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MIAO Lin, PENG Yanxia, WANG Feng, LI Yi, WANG Haowei, LI Lihua, ZENG Xiaofeng, YANG Genmeng
2023,33(7):1-8, DOI: 10. 3969 / j.issn.1671-7856. 2023. 07. 001
Abstract:
Objective To investigate damage of double-stranded DNA by methamphetamine (METH) in various types of nerve cells. Methods For in vitro experiments, primary cortical neurons and HT-22, BV2, HMC3 and U-87 MG cells were treated with METH. Cell morphology was observed and γ-H2AX expression was detected by Western blot. For in vivo experiments, METH was administered to mice by intraperitoneal injection, and behavioral analysis was performed by the open field test. Morphological changes of neurons in the prefrontal cortex and hippocampus of were observed by HE staining. Double-stranded DNA damage was observed by immunofluorescence, and γ-H2AX expression was detected by western blotting. Results In vitro, after treatment with METH, the morphology of primary cortical neurons, and HT-22, BV2, HMC3 and U-87 MG cells had obviously changed, the synapses of cells were shortened or disappeared, cell bodies had shrunk, and gaps were wider. γ-H2AX expression was significantly increased. In vivo, after administration of METH, compared with the saline group, the movement trajectory and distance were significantly increased in the METH group. HE staining showed that neurons in the prefrontal cortex and hippocampus were markedly edematous and eosinophilic, and some neurons had degenerated. Immunofluorescence showed that double-stranded DNA damage in neurons in the prefrontal cortex and hippocampus of the METH group was significantly increased, and the fluorescence intensity was significantly enhanced compared with the saline group. Western blot showed that γ-H2AX expression in the prefrontal cortex and hippocampus of the METH group was significantly increased compared with the saline group. Conclusions METH damaged doublestranded DNA in the nervous system. METH induced a significant increase in γ-H2AX expression in primary cortical neurons HT-22, BV2, HMC3 and U-87 MG cells, the prefrontal cortex, and hippocampus. This study provides a theoretical basis to elucidate the mechanism of METH-induced neurotoxicity.
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ZHANG Yixing, JIANG Zhiyan, LIU Anqi, WANG Shumin, SUN Chenghui
2023,33(7):9-16, DOI: 10. 3969 / j.issn.1671-7856. 2023. 07 002
Abstract:
Objective To evaluate the feasibility of an animal model of simulated adenoid hypertrophy by combining animal models of allergic rhinitis and chronic pharyngitis. Methods A rat model of allergic rhinitis was established by basal sensitization through intraperitoneal OVA injection and nasal enhanced sensitization. A model of chronic pharyngitis was established simultaneously by a throat spray of ammonia, both models were combined into a rat model of simulated adenoid hypertrophy. The symptom performance of model animals was assessed by the animal behavioral score after modeling. Pathomorphological changes of nasal and pharyngeal mucosae and nasopharyngeal lymph nodes were observed by HE staining. The percentage of eosinophils was assess in rat whole blood. IL-4 and IgE levels in rat serum and mucosal tissue were measured by ELISA. Results The behavior score, eosinophil percentage, and IL-4 and IgE levels in serum and tissue were increased compared with the control group (P< 0. 01). Compared with model group A, the eosinophil percentage and serum IgE level were increased in model group B (P<0. 01). In the model groups, nasal and pharyngeal mucosae and nasopharyngeal lymph nodes showed different degrees of disease-related histopathological changes. Conclusions The rat model had symptom manifestations and pathological changes similar to adenoid hypertrophy. This animal model of simulated adenoid hypertrophy can be used as a combined animal model of allergic rhinitis and chronic pharyngitis, but requires further exploration and improvement.
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ZHANG Gedi, LIU Gengxin, LUO Fuli, YAN Ziyou
2023,33(7):17-25, DOI: 10. 3969 / j.issn.1671-7856. 2023. 07. 003
Abstract:
Objective To predict the mechanism of Shenshuai Prescription (SSR) in chronic kidney disease(CKD)-related myocardial injury using network pharmacology and molecular docking method. Methods We used the traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP) and Herb herbal medicine identification database (http:/ / herb. ac. cn/ ), and the SwissTargetPrediction database to screen target information of active ingredients. We then used the UniProt database to screen for human targets and standard gene names. A drug active ingredient target network diagram was constructed using Cytoscape 3. 7. 2 software, and the GeneCards database was used to collect disease-related targets. The “ Shenshuai Recipe” against CKD myocardial injury gene target database was established using Venny 2. 1, and the STRING database was used to build the main component target interaction network and screen key targets. Cytoscape 3. 7. 2 software was imported for topology analysis and a protein-protein interaction network diagram was constructed. Finally, the DAVID platform was used for Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis and Gene Ontology (GO) biological function annotation. Results After screening, there were 252 active compounds in SSR and 649 common targets of SSR and CKD myocardial injury, among which AKT Serine/ Threonine Kinase 1 (AKT1), Tumor Necrosis Factor (TNF), Mitogen-Activated Protein Kinase 3 (MAPK3), and Vascular Endothelial Growth Factor A (VEGFA) may be important targets of SSR in treating CKD myocardial injury. GO analysis identified 1485 Biological process items, 176 Cell component items, and 386 Molecular function items, of which plasma membrane, cytosol, and cytoplasm had the largest number of enriched genes, and 313, 304, 276 genes were distributed respectively. KEGG analysis indicated that HIF-1α, Lipid and atherosclerosis, AGE-RAGE signaling path in diagnostic complexes, phosphoinositide 3-kinase (PI3K)-AKT, and insulin resistance pathways might be involved in the mechanisms of SSR in treating CKD myocardial injury. Conclusions SSR might play a role in cardiorenal protection by participating in multiple mechanisms, including improving insulin resistance, improving lipid metabolism, antiatherosclerosis, and regulating the expression of inflammatory factors and vascular endothelial growth factor, with the PI3K/Akt and mitogen-activated protein kinase pathways being potentially important signal regulation pathways.
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CHENG Jinlai, ZHOU Ziyu, LIU Li, SONG Lixia, HONG Kun, TAN Yuqing, YANG Miyi
2023,33(7):26-33,40, DOI: 10. 3969 / j.issn.1671-7856. 2023. 07. 004
Abstract:
Objective To investigate the factors affecting the establishment of a mouse model of alcoholic liver injury. Methods A total of 150 specific-pathogen-free male ICR mice were divided into three groups to study the possible influences of different experimental conditions, including modeling period, frequency, type, and dosage of modeling agent, and time interval between doses. Mice in the three groups received 60% ethanol and alcohol with 53% volume ratio by gavage at different doses, frequencies, and time intervals and the mortality rates were calculated. We also detected alanine aminotransferase (ALT) and aspartate transaminase (AST) serum levels and malondialdehyde (MDA), glutathione(GSH), and triglyceride (TG) levels in liver tissue homogenate. Liver pathology was examined in tissue sections. Results The mortality rates in the 1-day modeling and double-dose liquor modeling groups were 20% and 40%, respectively, and the mortality rates in the 4, 6, 8, and 12 h interval dosing groups were 40%, 20%, 10%, and 0%, respectively. Pathological indicators, including ALT, AST, MDA, GSH, TG, and tissue pathology worsened in line with shortening of the gavage interval and increased gavage dose. Conclusions The alcohol dose, length of the modeling period, and frequency of dosing affect the establishment of an alcoholic liver injury model in mice. We suggest that dosing mice with alcohol with 53% volume ratio twice a day at 20 mL/ kg body weight or 10 mL/ kg body weight at 6 h intervals can successfully cause liver damage.
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ZHAO Changcheng, DING Huaming, LU Lunshan, ZHANG Qian, YANG Jing, DONG Ying, XUE Qunshan, ZHUANG Ke
2023,33(7):34-40, DOI: 10. 3969 / j.issn.1671-7856. 2023. 07. 005
Abstract:
Objective To investigate the effect of HIV infection on inosine-5’-monophosphate dehydrogenase 2(IMPDH2) expression in the cerebral temporal cortex. Methods Montreal Cognitive Assessment (MoCA) scores and magnetic resonance imaging were used to assess mild cognitive impairment in HIV-infected patients. In addition, the histologic structure of the temporal cortex in a rhesus macaque with chronic simian-human immunodeficiency virus (SHIV) KU-1 infection was examined using hematoxylin and eosin staining. Moreover, IMPDH2 protein expression levels in paired healthy and infected temporal cortex were examined using immunohistochemistry and Western blot. Finally, putative genes associated with IMPDH2 expression were investigated using protein-protein interaction network analysis. Results HIV positive individuals had a thinner temporal cortex compared with healthy controls. In addition, the numbers of neurons in the temporal cortex were decreased and IMPDH2 protein levels were significantly lower in the infected monkey compared with healthy controls. Eight genes interacting with the IMPDH2 gene were identified. Conclusions Downregulation of IMPDH2 protein in the temporal cortex might be involved in the pathogenesis of HIV-related neurocognitive disease.
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LI Hongyan, MEI Xianyun, YANG Huiya, LI Yikang, FENG Pei, SUN Fangyun
2023,33(7):41-47, DOI: 10. 3969 / j.issn.1671-7856. 2023. 07. 006
Abstract:
Objective To investigate the effect and mechanism of Nardosinone (Nar) in hypoxia-induced apoptosis of H9c2 cardiomyocytes meditated through the PI3K/ Akt/ mTOR pathway. Methods Cobalt chloride (CoCl2 ) was used to establish a hypoxic injury model in H9c2 cardiomyocytes. Cell proliferation was assessed by CCK-8 assays. Hoechst 33342 was applied to counterstain cells, and laser scanning confocal microscopy was used to observe apoptosis. The potential signaling pathway of Nar in treating IHD was predicted by network pharmacology. Western blot was used to detect expression of apoptosis-, autophagy-, and PI3K/ Akt/ mTOR pathway-related proteins. Results The PI3K/ Akt/mTOR pathway is major signaling pathway in Nar treatment of IHD. Compared with the control group, CoCl2 (400 μmol/ L)decreased cell proliferation, the Bcl-2/ Bax ratio, PI3K, Akt and mTOR protein phosphorylation levels, and P62 protein expression, while increasing nuclear apoptotic fragmentation, the LC3II/ LC3I ratio, and Cleaved-Caspase-3 and Beclin-1 protein expression (P<0. 05). Compared with the CoCl2 group, Nar (50 μmol/ L) pretreatment significantly increased cell proliferation, alleviated nuclear apoptotic fragmentation, decreased Cleaved-Caspase-3 protein expression, increased the Bcl-2/ Bax ratio, and increased PI3K, Akt and mTOR phosphorylation levels and P62 protein expression. The LC3II/ LC3I ratio and the Beclin-1 protein expression were also decreased (P< 0. 05). Pretreatment with PI3K specific inhibitor LY294002 offset the effects of Nar on CoCl2-induced apoptosis, PI3K/ Akt/ mTOR pathway-related proteins, and P62 protein expression, but had no effect on the LC3II/ LC3I ratio or Beclin-1 expression. Conclusions Nar inhibited CoCl2-induced H9c2 cardiomyocyte apoptosis by activating the PI3K/ Akt/ mTOR pathway, but probably alleviated the damage caused by excessive autophagy in cells through other pathways.
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CHEN Wenpei, YANG Wei, DAI Jinlong, GUO Jianmin
2023,33(7):48-54, DOI: 10. 3969 / j.issn.1671-7856. 2023. 07. 007
Abstract:
Objective To establish and evaluate a chronic model of atopic dermatitis (AD) induced by 2,4-dinitrochlorobenzene (DNCB) in Brown Norway rats. Methods Brown Norway rats were randomly divided into negative, model, and tacrolimus groups. The ear skin of rats in model and tacrolimus groups were challenged by 0. 5% DNCB at days 1, 3, 7, 9, 12, 14, 16, 19, 21, 23 and 25. Rats in the control group were administered a substrate solution at the same volume at the same time points. Rats in the tacrolimus group were transdermally administered tacrolimus ointment for 25 collected and weighted after 26 days of modeling. Histopathological examination and toluidine blue staining of ear tissue were also performed. The IgE level in ear tissue was detected. Results The ear thickness in model rats was significantly increased after 9 days of challenge by DNCB, while whereas inflammation score was remarkedly improved after 12 days and reached a peak at 16~25 days. The weight of ear tissue and the IgE level in the ear were increased at day 26 of challenge. Pathological changes, such as epidermal thickening, inflammatory cell infiltration, and mast cell infiltration, occurred in the model group. Tacrolimus significantly reversed these changes. Conclusions Brown Norway rats challenged by DNCB develop stable AD-like symptoms, which are appropriate for pharmacological evaluation.
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DENG Yu, WEI Pixi, REN Shuangshuang, LYU Guixue, WEI Wenwen
2023,33(7):55-66, DOI: 10. 3969 / j.issn.1671-7856. 2023. 07. 008
Abstract:
Objective To explore the influence of miR-486-5p on the malignant behavior of endometrial cancer(EC) cells and its targeted regulation of PTEN. Methods The expression miR-486-5p and PTEN mRNA in EC tissues and cells was detected by qRT-PCR. The correlation between miR-486-5p and PTEN mRNA expression in EC tissues and the clinical characteristics of patients were analyzed. A dual luciferase reporter assay was performed to verify the relationship between miR-486-5p and PTEN. Ishikawa cells were separated into NC, NC inhibitor, miR-486-5p inhibitor, miR-486-5p inhibitor+si-NC, and miR-486-5p inhibitor+si-PTEN groups. Western blot was performed to detect expression of PTEN and PI3K/ AKT pathway-related proteins in cells. CCK-8 assays were performed to assess cell viability. Colony formation assays were used to examine the ability of cells to form colonies. Flow cytometry was performed to assess apoptosis, Scratch healing and transwell assays were applied to assess cell migration and invasion. Results The miR-486-5p was highly expressed in EC tissues and cells, while PTEN mRNA was expressed at a low level (P<0. 05). miR-486-5p and PTEN mRNA expression in EC tissues was related to the tumor size, differentiation degree, International Federation of Gynecology and Obstetrics stage, and lymph node metastasis (P<0. 05). miR-486-5p in Ishikawa cells might negatively target and regulate PTEN expression. Compared with NC and NC inhibitor groups, cell viability at 48 and 72 h, scratch healing rate, and ratios of p-PI3K/ PI3K and p-AKT/ AKT of Ishikawa cells in the miR-486-5p inhibitor group were decreased (P<0. 05), the number of colonies formed and the numbers of migrating and invasive cells were decreased (P<0. 05), and the apoptosis rate was increased (P<0. 05). Compared with miR-486-5p inhibitor and miR-486-5p inhibitor+si-NC groups, cell viability at 48 and 72 h, scratch healing rate, and ratios of p-PI3K/ PI3K and p-AKT/ AKT of Ishikawa cells in the miR-486-5p inhibitor+si-PTEN group were increased (P< 0. 05), the number of colonies formed and the numbers of migrating and invasive cells were increased (P<0. 05), and the apoptosis rate was decreased (P<0. 05). Conclusions Inhibition of miR-486-5p may suppress activation of the PI3K/ AKT pathway by negatively targeting PTEN, thereby inhibiting the proliferation, migration, and invasion of EC cells and inducing their apoptosis.
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JIANG Liping, WANG Xia, PENG Xinhua
2023,33(7):67-77, DOI: 10. 3969 / j.issn.1671-7856. 2023. 07. 009
Abstract:
Objective To explore how miR-34a-5p regulates insulin-like growth factor Ⅱ mRNA-binding protein 3 (IMP3) and programmed death ligand-1 (PD-L1) expression and its influence on the biological behavior of breast cancer cells. Methods We analyzed IMP3 and PD-L1 expression in breast cancer and normal tissues using the Human Protein Atlas and GEPIA, and patient prognosis and survival using The Cancer Genome Atlas and GTEx project. miR-34a-5p, IMP3, and PD-L1 mRNA levels, IMP3 and PD-L1 protein levels were detected in breast cancer and adjacent normal tissues by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blot. Correlations between miR-34a-5p and IMP3 and PD-L1 mRNAs were analyzed by Pearson’ s test. Immunohistochemistry was used to detect IMP3, PD-L1, and leukocyte differentiation antigen 44 variant 6 (CD44v6) expression in breast cancer and normal tissues, and the relationships between IMP3 and PD-L1 expression and clinicopathological parameters were analyzed. Target gene prediction and dual luciferase reporter assay were used to verify the targeted regulation of miR-34a-5p on IMP3 and PD-L1. MCF7 cells were divided into control group, miR-NC group, miR-34a-5p group, miR-NC+pcDNA-NC group, miR-NC+IMP3 group, miR-NC+PD-L1 group, miR-34a-5p+pcDNA-NC group, miR-34a-5p+IMP3 group, and miR-34a-5p+PD-L1 group. Cell proliferation, migration, and invasion were determined by MTT, colony-formation, scratch, and Transwell chamber tests, respectively. Expression levels of IMP3, PD-L1, CD44v6, matrix metalloproteinase-2 (MMP-2), MMP-9, and E-cadherin were detected by Western blot. Results The biological databases showed that higher expression levels of IMP3 and PD-L1 in breast cancer tissues were associated with shorter survival (P<0. 05). Expression levels of miR-34a-5p were significantly lower while expression levels of IMP3 and PD-L1 mRNAs and proteins were significantly higher in breast cancer tissues than in normal tissues. miR-34a-5p expression was negatively correlated with IMP3 and PD-L1 mRNAs, respectively (P<0. 05). Immunohistochemistry showed positive expression of IMP3, PD-L1, and CD44v6 in breast cancer, with higher IMP3 and PD-L1 expression associated with later TNM stage, lower degree of tumor differentiation, and increased risks of lymph node and distant metastases (P<0. 05). Dual luciferase assay verified the targeted regulation of miR-34a-5p on IMP3 and PD-L1. The cell viability, number of clones formed, scratch-healing rate, cell invasion, and expression levels of IMP3, PD-L1, CD44v6, MMP-2, and MMP-9 protein were all significantly lower and E-cadherin expression was significantly higher in the miR-34a-5p group compared with the control group and miR-NC group. The cell viability, number of clones formed, scratch-healing rate, cell invasion, and expression levels of IMP3, PD-L1, CD44v6, MMP-2, and MMP-9 proteins were all significantly increased while expression of E-cadherin protein was significantly decreased in the miR-NC+IMP3 and miR-NC+PD-L1 groups compared with the miR-NC+pcDNANC group. The cell viability, number of clones formed, rate of scratch healing, cell invasion, and expression levels of IMP3, PD-L1, CD44v6, MMP-2, and MMP-9 protein were all significantly increased while E-cadherin expression was significantly decreased in the miR-34a-5p+IMP3 and miR-34a-5p+PD-L1 groups compared with the miR-34a-5p+pcDNANC group (P< 0. 05). Conclusions miR-34a-5p can inhibit the proliferation, migration, invasion, and epithelialmesenchymal transition of breast cancer cells by targeting the expression of IMP3 and PD-L1.
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TIAN Lu, CHEN Ying, ZHANG Yang
2023,33(7):78-84, DOI: 10. 3969 / j.issn.1671-7856. 2023. 07. 010
Abstract:
Objective To investigate the effects of serum containing Fuzheng Kangai Decoction (FZKAD) on the proliferation, apoptosis, and cell cycle progression in human ovarian carcinoma HO-8910PM cells. Methods Forty Sprague-Dawley rats were divided randomly into control and low-dose (4. 725 g/ kg), medium-dose (9. 45 g/ kg), and high-dose (18. 9 g/ kg) FZKAD groups. After 7 days of intragastric administration, serum containing FZKAD was prepared and used for the culture of HO-8910PM cells. Cell proliferation was detected by MTT assay, colony-formation ability was detected by clone formation assay, apoptosis was detected by flow cytometry using Annexin V-fluorescein isothiocyanate/propidium iodide double-staining, cell cycle progression was detected by flow cytometry using propidium iodide single staining, and protein expression levels were detected by Western blot. Results There was no significant difference in cell survival rates among the low-dose, medium-dose, and control groups after 24 h ( P>0. 05). However, cell survival rates in the other groups were significantly decreased after 24, 48 and 72 h (P<0. 05 or P<0. 01), and the colony-formation abilities were also significantly decreased in each dosage group (P<0. 01). Compared with the control group, the apoptosis rates and protein expression levels of P53 and Bax were significantly increased in the low-dose, medium-dose, and highdose groups (P<0. 05 or P<0. 01), while the protein expression level of Bcl-2 was decreased (P<0. 01). The percentage of cells in G0 / G1 stage was significantly increased in the low-dose, medium-dose, and high-dose groups compared with the control group (P<0. 05 or P<0. 01), while the percentages of cells in S and G2 / M stages and protein expression levels of cyclin D1 and cyclin-dependent kinases 4 and 6 were decreased (P<0. 05 or P<0. 01). Conclusions Serum containing FZKAD can inhibit the proliferation of human ovarian cancer HO-8910PM cells, related to the induction of apoptosis and cell cycle arrest.
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LIU Yiqin, ZHU Tingting, MAO Haixia, KANG Ting, ZHANG Liling, WU Weihua, OU Santao
2023,33(7):85-91, DOI: 10. 3969 / j.issn.1671-7856. 2023. 07. 011
Abstract:
Objective To observe expression of ferroptosis-related factors in an adenine-induced chronic kidney disease model, and investigate the mechanism of ferroptosis in the pathological process of chronic kidney disease. Methods Twenty rats were randomly divided into a control group (Con) and model group (CKD). The CKD model was established by 2. 5% adenine gavage. All rats were sacrificed after 6 weeks. Blood samples were collected to measure serum urea nitrogen (BUN) and serum creatinine (Scr). HE staining was performed to observe renal pathological changes. Masson staining was performed to observe interstitial fibrosis of renal tubules. Iron, malondialdehyde (MDA), and glutathione(GSH) concentrations were measured. Western blot was performed to detect the protein expression of iron ferroptosisrelated proteins including systemic Xc- subunit SLC7A11, peroxidase 4 (GPX4), and iron metabolism-related proteins including transferrin receptor 1 (TFR-1), ferritin heavy chain (FTH), ferritin light chain (FTL), and membrane iron transporter protein (FPN). Correlations between iron, MDA and GSH levels and interstitial renal fibrosis were analyzed. Results Compared with the Con group, serum urea nitrogen and serum creatinine were significantly increased in the CKD group (P<0. 05). HE and Masson staining showed significant tubular dilatation and interstitial fibrosis. Iron and MDA concentrations were significantly increased (P<0. 05), and the GSH concentration was significantly decreased (P<0. 05). Immunohistochemistry showed that, 4-HNE protein expression was significantly increased and GPX4 protein expression was significantly decreased in the CKD group compared with the Con group. Western blot showed that SLC7A11, GPX4, TFR-1, and FPN protein expression was significantly decreased, while FTH and FTL expression was significantly increased compared with the Con group. Correlation analysis indicated that iron and MDA concentrations were positively correlated to the relative area (%) of renal fibrosis, and the GSH concentration was negatively correlated with them. Conclusions Ferroptosis is present in the adenine-induced CKD rat model and may be involved in renal fibrosis progression in CKD via the GSH-GPX4 axis.
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DU Chenfei, HAN Lei, FAN Wei, LI Hongqi, ZHENG Fuzeng
2023,33(7):92-99,122, DOI: 10. 3969 / j.issn.1671-7856. 2023. 07. 012
Abstract:
Objective To explore the effect of Rehmannia glutinosa polysaccharide (RGP) on apoptosis of osteoarthritis (OA) chondrocytes induced by sodium iodoacetate (MIA) via the regulation of long-chain non-coding RNA (lncRNA) maternal expression gene 3 (MEG3). Methods Rat chondrocytes were cultured with MIA 0, 1, 2, 4, and 8 μmol/ L to induce chondrocyte injury, and were then treated with RGP 50, 100, 200, 400, and 800 mg/ mL, respectively, to detect the appropriate experimental concentration. Rats were divided into a normal group, MIA group, RGP group, RGP+control (NC) small interfering RNA (siRNA) group, and RGP+si-MEG3 group. Cell viability was detected by Cell Counting Kit-8 (CCK-8) assay and apoptosis was detected by Hoechst 33258 staining and flow cytometry. mRNA levels of MEG3, metalloproteinase 13 (MMP-13), type Ⅱ collagen-α1 (COL2A1), and proteoglycan (ACAN) were detected by real-time quantitative polymerase chain reaction. Protein levels of PI3K, phospho (p) PI3K, serine/ threonine kinase(AKT), p-AKT, BAX, BCL-2, and caspase3 were detected by Western blot. Results MIA decreased chondrocyte viability and induced apoptosis in a dose-dependent manner, decreased MEG3, COL2A1, and ACAN mRNA levels, and increased MMP-13 mRNA levels (P<0. 05). RGP 100, 200, 400, and 800 mg/ mL increased chondrocyte viability and MEG3 levels (P <0. 05). Cell viability, MEG3, COL2A1, and ACAN mRNA, and p-PI3K/ PI3K, p-AKT/ AKT, and BCL-2 protein levels were decreased in the MIA group compared with the control group, while the apoptosis rate, MMP-13 mRNA, and BAX and caspase3 protein levels were increased (P<0. 05). Cell viability, MEG3, COL2A1, and ACAN mRNA, and p-PI3K/ PI3K, P-AKT/ AKT, and BCL-2 protein levels were increased in the RGP group compared with the MIA group, while the apoptosis rate, MMP-13 mRNA, and BAX and caspase3 protein levels were decreased (P<0. 05). Knockdown of MEG3 weakened the protective effect of RGP on MIA-induced chondrocyte injury. Conclusions RGP can promote the synthesis of chondrocyte extracellular matrix and inhibit cell apoptosis and MIA-induced chondrocyte damage, possibly acting via a mechanism related to the up-regulation of MEG3 expression and induction of PI3K/ AKT pathway activation.
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LI Dongdong, LI Xiaoming, WU Xuelian, YI Lu, CHEN Guangyou
2023,33(7):100-107, DOI: 10. 3969 / j.issn.1671-7856. 2023. 07. 013
Abstract:
Objective To investigate the therapeutic effect and mechanism of Gubi powder on osteoarthritis. Methods New Zealand white rabbits were divided into a blank group, model group, positive group (diclofenac potassium gel), and Gubi powder group (n = 6 rabbits per group). A rabbit knee osteoarthritis model was constructed using the modified Hulth method, and the treatments were administered for 4 weeks after 8 weeks of modeling. Cartilage thickness was measured by toluidine blue staining and the Mankin score was derived after Muscovite O staining of cartilage tissue. Terminal deoxynucleotidyl transferase dUTP nick end labeling fluorescence staining was used to detect the apoptosis of chondrocytes in cartilage tissue. Collagen (Col) Ⅱ and collagen X protein expression levels in cartilage tissue were detected by immunohistochemistry and expression levels of Bax, cleaved-caspase-3, phospho (p) extracellular signalregulated kinase (ERK), p-p38, p-Smad2, p-Smad3, and transforming growth factor (TGF)-β1 in cartilage tissues were detected by Western blot. Results Cartilage thickness was decreased, the Mankin score and chondrocyte apoptosis rate were increased, Col Ⅱ, p-Smad2, p-Smad3, and TGF-β1 protein expression levels were decreased, and Col X, Bax, cleaved-caspase-3, p-ERK, and p-p38 levels were increased in the model group compared with the control group. Cartilage thickness was increased, the Mankin score and chondrocyte apoptosis rate were decreased, Col Ⅱ, p-Smad2, p-Smad3, and TGF-β1 expression levels were increased, and Col X, Bax, cleaved-caspase-3, p-ERK, and p-p38 protein expression were decreased in the positive and Gubi powder groups compared with the model group. Conclusions Gubi powder showed good therapeutic efficacy against osteoarthritis in rabbits, by reducing chondrocyte apoptosis, stabilizing cartilage tissue structure, and promoting the regeneration and repair of cartilage tissue after injury. Its mechanism may be related to inhibition of the ERK/ P38 pathway and activation of the TGF-β/ Smad pathway.
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ZHANG Xiuyun, YANG Kun, LI Peijia, TANG Shun, YANG Siyu, CHEN Hui
2023,33(7):108-116, DOI: 10. 3969 / j.issn.1671-7856. 2023. 07. 014
Abstract:
Objective To explore the healing effect of xiaochaihu decoction in a rat model of chronic pyogenic osteomyelitis of the jaw. Methods Sixty-five specific-pathogen-free rats were used to establish a model of chronic pyogenic osteomyelitis of the jaw, by introducing Staphylococcus aureus into round bone defects prepared using a bone drill. After 1 month, five rats were randomly selected to test the modeling result, and the other rats were divided randomly into four groups: group A, debridement+saline; group B, debridement+xiaochaihu decoction; group C, debridement+cefuroxime; and group D, debridement+xiaochaihu decoction+cefuroxime. Five rats in each group were sacrificed at 2, 4, and 8 weeks after administration. The extent of the bone defect was detected by cone beam computed tomography (CBCT), levels of the inflammatory cytokines interleukin (IL)-6 and IL-1β were detected by enzyme-linked immunosorbent assay, and bone morphogenetic protein-2 was detected by immunohistochemistry. Results CBCT showed that the bone defects in groups B, C, and D had decreased at 2, 4, and 8 weeks after administration, compared with group A (P<0. 05). In addition, the bone defects were smaller in groups B and D compared with group C (P<0. 05), and smaller in group D compared with group B (P<0. 05). Levels of IL-6 and IL-1β were lower in groups B, C, and D compared with group A at 2, 4, and 8 weeks after administration, IL-1β levels were lower in groups B and D compared with group C, and levels of IL-6 were lower in group D compared with C (P<0. 05); however, there was no significant difference in IL-6 levels between groups B and C ( P>0. 05). IL-1β and IL-6 levels were both lower in group D compared with group B (P<0. 05). BMP-2 expression was detected in groups A, B, C, and D, with higher mean optical densities in groups B, C, and D compared with group A, higher optical densities in groups B and D compared with group C, and in group D compared with group B at 2, 4, and 8 weeks after administration (P< 0. 05). Conclusions We successfully established a rat model of chronic pyogenic osteomyelitis of the mandible using S. aureus. Xiaochaihu decoction improved chronic pyogenic osteomyelitis, and the therapeutic effect of xiaochaihu decoction combined with cefuroxime was better than that of either drug alone.
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ZHANG Wei, WANG Muzi, DENG Shaochang
2023,33(7):117-122, DOI: 10. 3969 / j.issn.1671-7856. 2023. 07. 015
Abstract:
Laboratory animal barrier contamination by pathogenic microorganisms during the operation of environmental facilities seriously affects research and animal health or leads to biosafety incidents, interference of experimental result, or interruption of experiments. However, elimination of contamination during their long-term operation is difficult ,and can occur due to careless management or operational details. Contamination by pathogenic microorganisms is a major problem for laboratory animal managers. After engaged in laboratory animal barrier facility management for many years, The author deeply felt the importance of details management to prevent infection by pathogenic microorganism. Therefore, summarized of the details problem combined facility layout with staff management based on years of working experience, which in order to be discussed and used for reference.
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PENG Mengwei, LIU Yan, ZHANG Yan, HAO Linglun, CHEN Yulong, WU Yaosong
2023,33(7):123-129, DOI: 10. 3969 / j.issn.1671-7856. 2023. 07. 016
Abstract:
Cachexia is a syndrome of multifactorial metabolic disorders involving multiple organs and is a common fatal factor in patients with malignant tumors. The major clinical symptoms are muscle atrophy and loss, and their mechanism is associated with excessive protein degradation and synthesis disorders. In recent years, traditional Chinese medicine has made a significant advantage in the use of few side effects, multi-targets, and multi-channel intervention diseases. It has unique value in preventing and treating tumor disease and muscle atrophy. This review mainly discusses the effect of muscle protein degradation and synthesis mediated by various signals on tumor Cachexia muscle atrophy and the regulatory role of traditional Chinese medicine to provide a reference for the clinical application and research of traditional Chinese medicine in the treatment of tumor Cachexia muscle atrophy.
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TIAN Jing, MA Yingjie, WANG Peng, LI Yang, ZENG Lu, LI Bin, SU Guohai
2023,33(7):130-140, DOI: 10. 3969 / j.issn.1671-7856. 2023. 07. 017
Abstract:
Vascular endothelial cells form a thin layer of flattened cells that cover the surface of the vessel wall, As an effector tissue, the vessel wall responds to various stimuli to release vasoactive substances to maintain homeostasis. However, endothelial cells are prone to dysfunction induced by multiple pathological factors, which is manifested by an impaired vasomotor function, thrombosis, and arterial wall proliferation. Studies have shown that vascular endothelial dysfunction is closely related to the occurrence and development of cardiovascular diseases such as atherosclerosis and heart failure. Therefore, effectively evaluating endothelial functions is essential to prevent and treat cardiovascular diseases. In this review, we discuss the functions and regulatory principles of the vascular endothelium and its relationship with cardiovascular diseases and summarize the basic principles and advantages and disadvantages of existing endothelial assessment techniques to assist in the diagnosis and treatment of cardiovascular diseases.
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WANG Baoyue, WANG Yongfu, SUN Xiaolin
2023,33(7):141-148, DOI: 10. 3969 / j.issn.1671-7856. 2023. 07. 018
Abstract:
Accumulating studies have shown that long non-coding RNA (lncRNA) plays major roles in endothelial injury, inflammation, cancer, and autoimmune diseases. Systemic sclerosis (SSc) is an autoimmune disease with an unclear pathogenesis. lncRNA may be involved in the occurrence and development of SSc by regulating immune disorders, microvascular disease, and fibrosis, and may be a biomarker of SSc to predict occurrence, activity and progression of the disease. Therefore, lncRNA may be a novel therapeutic target of SSc.
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SONG Yagang, TIAN Shuo, QIAO Jingyi, WU Xiangxiang, ZHU Xinghao, LIU Tiantian, MIAO Mingsan
2023,33(7):149-154, DOI: 10. 3969 / j.issn.1671-7856. 2023. 07. 019
Abstract:
Toxicity has become a major obstacle to the international use of traditional Chinese medicines, and a rational grading strategy for their toxicity is therefore needed. Although the existing traditional toxicity classification can help to guide the clinical use of traditional Chinese medicines, it lacks quantitative data and is not sufficiently accurate, with different sources reporting different toxicity classifications for the same drug. Modern research on toxicity classification mainly includes classification method based on LD50 of oral raw drugs, multi indicator comprehensive classification method, evaluation model of toxic traditional Chinese medicine based on “substance efficacy toxicity”, and the use of modern toxicology research models. Although these toxicity grading method achieve quantitative standards for traditional Chinese medicine toxicity grading, they deviate from the theoretical guidance of traditional Chinese medicine and differ significantly from clinical practice, failing to reflect the essence of clinical compound medication of traditional Chinese medicine. To sort previous studies on the toxicity of traditional Chinese medicines, this paper proposes a scoring and grading method based on data mining combined with toxicological studies. The proposed method is based on data mining of historical toxicity records for traditional Chinese medicines, clinical reports, and experimental in vitro and in vivo studies, as well as comprehensive modern toxicological research result, followed by the assignment of weighted scores to each item, to derive a toxicity classification method for traditional Chinese medicines. For example, aconite would be graded as highly toxic according to this procedure. This grading method has the advantages of focusing on clinical compound drug use, as well as providing guidance for clinical practice. It can quantitatively indicate the degree of toxicity, thus reflecting the modern characteristics of traditional Chinese medicine. This grading method needs further refinement, but after gradual improvement and refinement, this toxicity classification method for traditional Chinese medicines is expected to promote their rational clinical application and wider international use.
Volume 33,2023 Issue 7
Volume 33,2023 Issue 7
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The expression of IGHG1 in acute myeloid leukemia THP-1 cell and the influences on cell proliferation, apoptosis and invasion by regulating TGF-β/Smad pathway
Abstract:
Objective To investigate the expression of immunoglobulin γ-1 heavy chain constant region (IGHG1) in acute myeloid leukemia (AML) THP-1 cell and its influences on cell proliferation, apoptosis and invasion by regulating the transforming growth factor β (TGF-β)/Smad pathway. Methods The bone marrow specimens of 9 children with AML, the bone marrow specimens of 8 children with fracture,human bone marrow stromal cells HS-5 and human AML cells THP-1, HL60 were used as research objects,Western Blot was used to detect IGHG1 protein expression; THP-1 cells were divided into: blank group (cells without any treatment), si-NC group, si-IGHG1-1 group, si-IGHG1-2 group, si-IGHG1-3 group, TGF-β group,si-IGHG1-1 TGF-β group,si-IGHG1-1 TGF-β LY364947 group, CCK-8 method was used to detect cell proliferation; apoptosis was detected by flow cytometry; Transwell experiment was used to detect cell invasion ; Western blot was used to detect the protein expression of IGHG1, TGF-β, p-Smad2, and p-Smad3 in each group of cells. Results Compared with the bone marrow of children with fracture, the expression level of IGHG1 protein [(0.24?.03) vs (0.87?.12] in the bone marrow of children with AML was significantly higher (P<0.05); compared with HS-5 cell, the expression level of IGHG1 protein in human AML cells THP-1, HL60 was significantly increased [(0.89?.14)(0.75?.08) vs (0.21?.02)] (P<0.05); compared with the blank group , the OD450 value (24, 48, 72h) of THP-1 cells, the number of invaded cells, and the protein expression of TGF-β, p-Smad2, and p-Smad3 were significantly reduced in the si-IGHG1-1 group, and the apoptosis rate was increased (P<0.05), while the corresponding indexes in TGF-β group were opposite (P<0.05); the TGF-β reversed the effects of silencing IGHG1 on the proliferation, apoptosis and invasion of THP-1 cells; compared with si-IGHG1-1 TGF-β group, TGF-β, p-Smad2, p-Smad3 protein, OD450 values (24, 48, 72 h) and invasion number of cells decreased significantly in si-IGHG1-1 TGF-β LY364947 group, and the cell apoptosis rate was increased (P<0.05). Conclusions IGHG1 is highly expressed in AML cells. Silencing IGHG1 can inhibit the proliferation and invasion of AML cells, and promote the apoptosis of AML cells. This mechanism may be related to the inhibition of TGF-β/Smad pathway.
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The relationship between atmospheric pollutants and some adverse pregnancy outcomes
jinjian, Chang Hui, Huang Hehuan, Peng Rui, Zhang Xiaoan
Abstract:
With the rapid development of China's economy and the continuous acceleration of urbanization, the problem of air pollution is becoming increasingly prominent, which has a great impact on human health. In recent years, a large number of research work has emerged both domestically and internationally, and research results have shown a certain correlation between exposure to atmospheric pollutants during pregnancy and adverse pregnancy outcomes. The impact of air pollution on the health and embryonic development of pregnant women has received more attention. This article mainly provides a brief review on the impact of atmospheric pollutants on some adverse pregnancy outcomes and their possible biological mechanisms.
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Research Progress on Model Organism Zebrafish in Neurodegenerative Diseases
Huang Kongli, Su Shijie, Wang Yuting, LIU ZHUANGZHUANG, CAO DANDAN, WANG PEI, CHEN GUANLIN, Wang Qi
Abstract:
Neurodegenerative disease （NDD）is a chronic and progressive neurological disease characterized by the loss of a large number of specific neurons. It mainly includes Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD) and amyotrophic lateral sclerosis (ALS). Although the lesions and etiologies of different types of neurodegenerative diseases are different, delayed neurodegenerative lesions and cell loss in specific brain areas are their common characteristics. For this reason, it is collectively referred to as neurodegenerative diseases. In recent years, zebrafish have attracted increasing attention from society as a new type of model organism. Although there are some differences between zebrafish and human central nervous systems, zebrafish neural conduction systems, neuronal and glial cell types, and disease-related gene homology are very similar to those of humans. Zebrafish has been widely used in the study of neurodegenerative diseases, and some achievements have been made in this field to improve our understanding of these diseases, but because of the complexity, multi-factors and multi-gene regulation of neurodegenerative diseases, the etiology and pathogenesis of zebrafish are not clear, so the treatment of this kind of disease has always been a difficult problem. By consulting relevant literature from home and abroad in recent years, this study reviews recent advances in neurodegenerative diseases using zebrafish as a model organism.
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Progress in irisin and its upstream and downstream antidepressants
Sangjiala, Lishanshan, Cuixin, Renqingqing, Houruiling, Panxingfang, Wangshenjun, Zhao
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Depression is the main cause of disability, and the adverse effects on people are difficult to eliminate. Despite the increasing number of antidepressants, depression is still not adequately treated clinically, and new mechanisms still need to be explored. As the beneficial effects of irisin on the nervous system are gradually elucidated, studies have found that irisin has an antidepressant effect, and irisin may become a new target for the treatment of depression. This study aims to explore the mechanism of irisin and its upstream and downstream antidepressants, by reviewing the existing studies to explain the link between depression and irisin, propose the potential mechanism of SIRT1 / PGC-1α mediating FNDC5/Irisin to regulate BDNF and promote neurogenesis to improve depression, and provide a new idea for the study of irisin and its upstream and downstream antidepressants.
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Advances of CREB in the regulation of memory through synaptic plasticity and its relation with Alzheimer’s disease
Luo Zhuohui, pang shuo, Zhang lianfeng
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Alzheimer’s disease is an irreversible heterogeneous neurodegenerative disease. AD patients are featured with memory loss and impaired synaptic plasticity. With a view to cAMP responsive element-binding protein which is intimately associated with synaptic plasticity. this assay summarized progress on the structure, signal pathway, downstream genes and relative memory regulation. The involvement of CREB in AD development would serve as references for AD researches based on synaptic plasticity improvement.
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Research progress on the regulation of lymphangiogenesis and its role in myocardial infarction
Abstract:
Lymphatic system plays an important role in regulating interstitial fluid homeostasis, lipid metabolism and immune function. After myocardial infarction, enhanced lymphangiogenesis accelerates the clearance of infiltrating immune cells, reduces the production of pro-inflammatory cytokines, reduces edema, inflammation, and fibrosis, and promotes the recovery of damaged heart function. Vascular endothelial growth factor-C (VEGF-C) and its receptor VEGFR-3 are components of the lymphangiogenesis pathway and play a critical role in maintaining tissue fluid balance and myocardial function after cardiac injury. Lymphatic vessels are closely related to the immune system. Different immune cell groups can stimulate or inhibit lymphatic remodeling. Macrophages are congenital immune cells widely distributed in organs and tissues, and play an important role in various physiological and pathological processes such as organ development, host defense, acute and chronic inflammation, tissue homeostasis and remodeling. More mechanisms of lymphangiogenesis need to be identified to provide effective targets for clinical stimulation of lymphangiogenesis to treat heart disease. This paper reviews the basic pathological changes of the heart and lymphatic vessels after myocardial infarction, the regulatory factors of lymphangiogenesis, and the influence of macrophages on lymphangiogenesis.
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Effects of LncRNA FGD5-AS1 targeting and regulating miR-129-5p on the proliferation, apoptosis, migration and invasion of oral squamous cell carcinoma cells
Abstract:
Objective To investigate the effects of long non-coding RNA (LncRNA) FGD5-AS1 on the proliferation, apoptosis, migration and invasion of oral squamous cell carcinoma (OSCC) cells through targeted regulation of miR-129-5p. Methods The expression of FGD5-AS1 in OSCC was analyzed by online database. The tumor tissues, normal tissues, and the human oral mucosal cells (HOK) and OSCC cells (SCC-9, HSC-4, SCC-25, CAL-27) cultured in vitro from 30 OSCC patients collected in our hospital were used as research subjects, qRT-PCR method was performed to detect the expression of FGD5-AS1 and miR-129-5p. The CAL-27 cell line with the highest FGD5-AS1 expression was randomly separated into Control group (normal culture, no transfection), si-NC group (transfected with si-NC), si-FGD5-AS1 group (transfected with si-FGD5-AS1), si-FGD5-AS1 NC inhibitor group (co-transfected with si-FGD5-AS1 and NC inhibitor) and si-FGD5-AS1 miR-129-5p inhibitor group (co-transfected with si-FGD5-AS1 and miR -129-5p inhibitor), CCK-8 method and clone formation assay were used to detect the proliferation ability of CAL-27 cells; the apoptosis level of CAL-27 cells was detected by flow cytometry; the migration ability of CAL-27 cells was detected by wound-healing assay; Transwell chamber was used to detect the invasion ability of CAL-27 cells; and dual luciferase reporter experiment verified the targeting relationship between FGD5-AS1 and miR-129-5p; the expression of high mobility group protein B1(HMGB1) was detected by Western blot. In vivo xenograft tumor model was constructed and divided into sh-NC group, sh-FGD5-AS1 group, miR-129-5p inhibitor group, and sh-FGD5-AS1 miR-129-5p inhibitor group. Tumor volume and tumor were detected. QRT-pcr was used to detect the expression of FGD5-AS1 and miR-129-5p in transplanted tumor tissues. The expression of HMGB1 and Ki67 was detected by immunohistochemistry. Results Database analysis showed that the expression level of FGD5-AS1 in OSCC tumor tissues was 4 times higher than that in normal tissues, and FGD5-AS1 expression was associated with poor grade in OSCC patients. Compared with normal tissues or human oral mucosal cells, the expression of FGD5-AS1 in tumor tissues and OSCC cell lines was significantly increased, and the expression of miR-129-5p was significantly decreased (P<0.05), the CAL-27 cells with the highest expression level of FGD5-AS1 were selected for transfection experiments. Compared with the Control group and the si-NC group, the apoptosis rate of the si-FGD5-AS1 group was significantly increased, and the OD value (48 h, 72 h, 96 h), scratch healing rate and the number of invaded cells were significantly reduced (P<0.05). MiR-129-5p was the target gene of FGD5-AS1. Inhibiting the expression of miR-129-5p was able to reverse the effects of interference FGD5-AS1 on the proliferation, apoptosis, migration and invasion of OSCC cells, thereby restoring the cancer-promoting effect of FGD5-AS1. After FGD5-AS1 was disrupted, HMGB1 expression was down-regulated by significantly enhancing miR-129-5p expression (P<0.05). In vivo experiments showed that FGD5-AS1 silencing significantly inhibited the growth and expression of HMGB1 and Ki67 (P<0.05), inhibition of miR-129-5p was the opposite; Inhibition of miR-129-5p reversed the inhibition of FGD5-AS1 on tumor growth and expression of HMGB1 and Ki67 (P<0.05). Conclusions FGD5-AS1 is up-regulated in OSCC cells. Interfering with FGD5-AS1 can inhibit proliferation, migration and invasion of OSCC cells and promote apoptosis by targeting miR-129-5p / HMGB1 axis.
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Behavioral assessment in the parkinson’s model of nonhuman primates
Abstract:
Parkinson's disease is a neurodegenerative disease, can cause a series of patients with symptoms of movement and the movement, behind the pathogenesis is unclear, to explore the mechanism behind these phenotypic requires an appropriate animal model of rodents in the movement and the movement is very difficult to accurately simulate the patient symptoms, causes related to preclinical studies have limitations, in translational medicine. Non-human primate animal models can make up the gap between rodents and humans, thus the non-human primate PD model of movement and the movement symptoms, is very important to research on mechanism and therapy, this review summarizes the different behavior in quantitative way, and compared the advantages and disadvantages between different methods, for studying the PD monkey model provides a behavioral testing for help.
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Autophagy: the key mechanism of exercise in improving neurodegenerative diseases
Chen Xianghe, Qiu Xiao, Liu Chi, Shen Ziming, Zhou Xiangxiang
Abstract:
Autophagy regulation of neurological diseases is the focus of current research in the field of neuroscience. Autophagy disorder leads to protein expression, deposition and dysfunction such as Aβ、Tau、α-syn and causes neurodegenerative diseases such as AD, PD and HD. Exercise is an important means to improve neurodegenerative diseases, which is closely related to the up-regulated expression of LC3, Beclin-1, Lamp1 and other autophagic factors after the activation of AdipoR1/AMPK/TFEB, AMPK/mTOR and other pathways. Higher autophagy level can remove the deposition of Aβ、Tau、α-syn and other proteins in the brain and can improve the neurodegeneration, synaptic structure and function disorder caused by neurodegenerative diseases. This study reviewed and analyzed the mechanism of autophagy in the improvement of neurodegenerative diseases by exercise, which will provide a solid theoretical basis and new research ideas for the improvement of neurodegenerative diseases by exercise.
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Advances in the construction of animal models of severe asthma
Wei Dandan, Wang Liting, Long Jie, Chen Yanjiao, Wang Yu, Yang Yongqing, Xu Yudong
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Severe asthma (SA), which requires high doses of glucocorticoids in combination with other medications to maintain symptom control or is uncontrollable even with these treatments, is currently a challenge in the clinical management of asthma. The establishment of a stable and reproducible experimental animal model that highly mimics the pathophysiological and clinical characteristics of patients with severe asthma is a key fundamental link in deepening the study on the pathogenesis of severe asthma, identifying potential therapeutic targets and developing targeted drugs. This paper reviews the experimental studies related to the construction of animal models of severe asthma in the past 10 years, and summarizes the recent progress in the establishment and evaluation of animal models of severe asthma from three aspects: animal selection, animal model construction protocols, and model pathological phenotypes. To summarize and analyze the latest progress in the construction of SA animal models, and provide a reference for SA basic research based on animal models.
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Research progress in experimental models of pulmonary arterial hypertension
xiejincheng, Chenjianying, Dengshaodong, Xiao mengyuan
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Pulmonary hypertension is a chronic progressive disease that, if not treated promptly, can ultimately lead to right heart failure and even death In order to explore the pathogenesis of pulmonary arterial hypertension and find more effective treatment methods, it is necessary to establish a suitable experimental model. In this paper, Pubmed, CNKI and other databases were used to search relevant literature, and the preparation methods and research progress of various experimental models of pulmonary arterial hypertension in recent years were summarized. The preparation methods of the models were classified to show the pathological characteristics of the disease more directly. On this basis, the present situation and existing problems of the in vitro and in vivo model of pulmonary arterial hypertension were discussed, in order to provide reference for the pathogenesis and clinical research of pulmonary arterial hypertension.
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Research progress of experimental premenstrual dysphoria model in rodents
Abstract:
Premenstrual dysphoric disorder (PMDD) is a subtype of Premenstrual syndrome with high prevalence in women during their reproductive life. It includes physical symptoms such as breast tenderness, headache as well as serious emotional symptoms such as anxiety, depression and irritability. These symptoms are menstrual specific, they appear before menstruation and disappear within a week after it ends. PMDD threatens the health of female patients all over the world, In The Diagnostic and Statistical Manual of Mental Disorder(Fifth edition ed,DSM-5）, PMDD is identified as one of the five types of depression. At present, studies on the pathological mechanism of PMDD mainly focus on hormone and receptor expression, which is the theoretical basis of the animal model establishment, as well. Animal model that can model after clinical symptoms of human beings is a very important to connect basic research to clinical research, which is also crucial to explore the pathogenesis and develop appropriate drugs. Although many kinds of animal models of PMDD have been used in experimental studies, there are still many limitations, which resulting in a barrier to further investigation of PMDD. Therefore, this review arranged the known pathogenesis research of PMDD and discuss the existing animal models of PMDD, aims to provide reference for PMDD related mechanism research and drug development.
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Analysis of animal models of keratitis based on the characteristics of clinical conditions of traditional Chinese and Western medicine
Abstract:
Abstract: Keratitis is a common ophthalmic disease with a high rate of blindness. This paper reviews a large number of relevant literature of animal models of keratitis, summarizes and analyzes the mechanism of existing animal models of keratitis, and evaluates the clinical anastomosis degree of traditional Chinese and Western medicine in combination with the established clinical diagnostic standards of traditional Chinese and Western medicine, analyzes its advantages and disadvantages, and puts forward some suggestions. It is found that most of the existing models are based on Western medical theories, and lack the diagnostic standards of traditional Chinese medicine. The model is mostly modeled by a single factor, which cannot well simulate the real lesion process of the disease; The diagnosis of the model lacks apparent indicators; It is recommended that animals be modeled by combining disease evidence. It is very important to prepare animal models of keratitis with high clinical compatibility and establish perfect evaluation standards, which can help us better understand the occurrence and development of keratitis in modern medical experiments, and effectively prevent and treat it.
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Study on the uptake and transport properties of euscaphic acid in Caco-2 cell model based on UPLC-TQ-MS
wangning, xieli, ding wenhuan, tian li
Abstract:
ABSTRACT: Objective Caco-2 cell model was established to explore euscaphic acid uptake and transport mechanism in this study. Methods Ultra-high performance liquid chromatography-triple quadrupole-mass spectrometry(UPLC-TQ-MS) was employed to determine the content of euscaphic acid. The effects of different time and temperature on its uptake were investigated. Based on the results of the uptake studies, the effects of different concentrations, P-gp inhibitors, chelating agents and pH values on its bidirectional transport were explored. Results The uptake of euscaphic acid was (8.38 ?0.87) μg穖g-1 for 180 min on the Caco-2 cell model at 37 癈. The apparent permeability coefficient (Papp values) of euscaphic acid for low, medium and high concentrations were(61.41 ?.92)?0-4、(146.90?4.91)?0-4 and (167.18?.72)?0-4 cm穝-1 respectively, which were positively correlated with the concentrations. P-gp inhibitors and chelating agents had no effect on its Papp values. Weakly acidic environment (pH6.00) significantly increased its Papp value, and the efflux rate (ER) ranged from 0.8–1.4. Conclusion The above results indicate that euscaphic acid has good transmembrane permeability in Caco-2 cell model, and the uptake mode is mainly passive diffusion. It is not a substrate of P-gp and there is no cellular bypass transport. This research may provide an experimental basis for the in vivo intestinal absorption of medicines containing euscaphic acid.
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Mechanism of taxifolin regulating endoplasmic reticulum stress PERK-ATF4 pathway to reduce myocardial hypertrophy in hypertensive rats
yujinyu, hanjing, zhangying, yuwen
Abstract:
【Abstract】 Objective To investigate the impact and molecular mechanism of taxifolin (TAX) on myocardial hypertrophy in spontaneously hypertensive rats (SHR). Methods Twenty-four SHRs were separated into SHR control group (SHR group), TAX group (20 mg/kg), and TAX PERK activator CCT020312 (CCT) group (20 mg/kg TAX 2 mg/kg CCT), 8 per group; another 8 normal blood pressure Wistar-Kyoto (WKY) rats were regarded as the normal control group (WKY group), and all were given corresponding drugs for 8 weeks of continuous intervention. During the experiment, the changes in blood pressure of the rats were observed, and after the intervention, the thickness of the diastolic ventricular septum (IVSd), the thickness of the systolic ventricular septum (IVSs), and the left ventricular ejection fraction (LVEF) were detected by echocardiography to determine the degree of myocardial hypertrophy and cardiac function, then the cardiac index and left ventricular index were calculated, hematoxylin-eosin (HE) staining, wheat germ agglutinin (WGA) staining and Masson staining were performed to evaluate the pathological changes of myocardial tissue, real-time quantitative PCR (qRT-PCR) was performed to detect the expressions of atrial natriuretic peptide (ANP), B-type natriuretic peptide (BNP), type I collagen α1 chain (COL1A1) and type III collagen α1 chain (COL3A1) mRNA in myocardial tissues, Western blot was performed to detect the expressions of protein kinase R-like endoplasmic reticulum kinase (PERK)-activator of transcription 4 (ATF4) pathway-related proteins in cardiac muscle. Results After the intervention, compared with the WKY group, the systolic blood pressure (SBP), diastolic blood pressure (DBP), IVSd, IVSs, cardiac index, left ventricular index, myocardial cell cross-sectional area, collagen volume fraction (CVF), myocardial tissue ANP, BNP, COL1A1 and COL3A1 mRNA expressions, glucose-regulated protein 78 (GRP78), ATF4, C/EBP homologous protein (CHOP) protein levels and p-PERK/PERK ratio in the SHR group increased (all P<0.05), LVEF decreased (P<0.05); compared with SHR group, the SBP, DBP, IVSd, IVSs, cardiac index, left ventricular index, myocardial cell cross-sectional area, CVF, myocardial tissue ANP, BNP, COL1A1 and COL3A1 mRNA expressions, GRP78, ATF4, CHOP protein levels and p-PERK/PERK ratio in TAX group decreased (all P<0.05), LVEF increased (P<0.05); CCT020312 partially reversed the protective effects of TAX on cardiac function and cardiac hypertrophy. Conclusion TAX can improve hypertensive myocardial hypertrophy by inhibiting endoplasmic reticulum stress (ERS), and its mechanism may be related to inhibiting PERK-ATF4 pathway.
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Study on tissue distribution difference of ligustrazine-Salvia miltiorrhiza before and after compatibility in acute myocardial ischemia model rats
lirong, goujian, liuting, gongzipeng, luyuan, liuchunhua, hunagyong, sunjia
Abstract:
Objective To investigate the difference in tissue distribution of ligustrazine hydrochloride and Salvia miltiorrhiza (Danshen) in Acute myocardial ischemia (AMI) rats. Methods AMI model was induced by subcutaneous injection of isoproterenol hydrochloride. UPLC-MS/MS method was used to determine the content differences of ligustrazine and danshensu in heart, liver, spleen, lung, kidney and brain tissues after single intravenous injection of Danshen injection (DGI), Ligustrazine injection (CGI) and Shenxiong glucose injection (SGI) at different time points. Results After intravenous injection, ligustrazine and danshensu were widely distributed in tissues of AMI rats and reached their peak rapidly. Before and after compatibility, the content of ligustrazine was the highest in brain tissue and the lowest in liver tissue. The content of danshensu in kidney tissue was the highest, and in brain tissue was the lowest, indicating that ligustrazine could penetrate the blood-brain barrier more easily than danshensu, and the main accumulation tissues were different. The AUC of ligustrazine and danshensu in the heart of the target organ was significantly increased (P < 0.001). Conclusion It is speculated that the combination of ligustrazine and salvia miltiorrhiza may enhance the therapeutic effect by increasing the distribution of main pharmacodynamic components in the heart, so that SGI can play a better role.
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The role of PI3K/Akt/mTOR signaling pathway in the pathogenesis of TRALI rats
liushuangchun, Zhang Xijiang, Chen Zaihuan, Wang Luqian, Lin Hairong
Abstract:
Objective: To study the effect of PI3K/Akt/mTOR signaling pathway on the pathogenesis of TRALI. Methods: Animal model was established by the method of trauma-blood loss-massive transfusion, and the pulmonary histopathological changes were detected by HE staining to determine whether the rats had pulmonary edema. The protein and mRNA expression levels of TNF-α, IL-6 and IL-1β in peripheral blood or lung tissues in TRALI rat models were detected by ELISA and qRT-PCR. The expression levels of PI3K/Akt/mTOR signaling pathway related proteins and apoptosis-related proteins Bax, Bcl-2 and Caspase3 were detected by western blotting. Results: Alveolar tissue structure was seriously damaged, the alveolar wall was thickened, there was pink edema fluid in the alveolar cavity, inflammatory cells infiltrated, edema was obvious in TRALI model rat. The expression levels of inflammatory cytokines TNF-α, IL-6 and IL-1β were significantly increased in peripheral blood and lung tissues (P<0.05); PI3K/Akt/mTOR signaling pathway was activated, the expression of p-mTOR/mTOR was significantly increased, and the expression of apoptotic protein Bax and Caspase 3 was inhibited, and the expression of anti-apoptotic protein Bcl-2 was increased (P<0.05). Conclusion: As a potential drug target, mTOR is an important means for clinical prevention and control of the occurrence and development of TRALI by defining the exact time target of its protective and damaging effects and selecting the optimal time of medication due to its complicated mechanism.
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Alterations of Gut Microbiota - Short Chain Fatty Acid Axis in IBS-D Rats and the Effect of Sodium Butyrate
zhan kai, Wu haomeng, zheng huan, qin shumin, Huang shaogang, yang yuanming
Abstract:
Objective To explore the alterations of gut microbiota - short chain fatty acid metabolic axis in IBS-D rat model, and to explore the effect of sodium butyrate on this axis. Methods Seventeen Wistar rats were randomly divided into three groups. The IBS-D rat model was induced by 4% acetic acid enema combined with restraint stress. During the modeling period, sodium butyrate was injected intraperitoneally. The fecal flora of rats was detected by 16sRNA technology, and the fecal short chain fatty acid content of rats was detected by liquid chromatography mass spectrometry (LC-MS). Results Compared with the normal group, the weight of the model group was decreased, AWR score and fecal water content have a significant increase, the relative abundance of intestinal probiotics was decreased，while that of g-Blautia was elevated, the fecal acetic acid content was increased significantly, and the contents of butyric acid and valeric acid was decreased significantly in the model group. After drug intervention, compared with the model group, the weight of rats in the sodium butyrate group was increased, the AWR score and fecal water content was decreased significantly, the relative abundance of g-Blautia was decreased significantly, and the fecal butyric acid and valeric acid content was obviously increased. Conclusions Sodium butyrate can alleviate diarrhea and colon visceral hypersensitivity in IBS-D rats by improving the imbalance of gut microbiota - short chain fatty acid metabolic axis.
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Advances in Ferroptosis-Based Therapy for Aging and Aging-Related Diseases
Zhou yongchang, Tao Siman, Chen Shengqiang, Pu Xiuying
Abstract:
Ferroptosis is a novel form of programmed cellular necrosis proposed in recent years, which is found in various diseases and seriously impacts on human health. Numerous studies have shown that inhibition or promotion of ferroptosis can improve many aging and aging-related diseases, and Traditional Chinese medicine plays an important role in this regard. This paper reviews the effects and mechanisms of Ferroptosis on aging and aging-related diseases, and compares the research progress of TCM in the treatment of ferroptosis-mediated aging and aging-related diseases in recent years, aiming to provide more options for the treatment and prevention of aging and aging-related diseases.
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Research progress in the pathogenesis of inflammatory bowel disease
Houxiaoting, Menghuan, Xuejiachen, Zhanghuamin, Wangkesi, Wangxude
Abstract:
Inflammatory bowel disease (IBD) is a global idiopathic disease, involving the ileum, rectum and colon. IBD begins to appear early in clinical symptoms, including abdominal discomfort, diarrhea, hematochezia, fever, fatigue and weight loss. The diagnosis of IBD is based on the comprehensive evaluation of relevant clinical manifestations, endoscopic examination results and histopathological characteristics of pathological tissue specimens. Biological agents, oral corticosteroids, salicylic acid and surgery are the main conventional treatments for IBD. In recent years, considerable progress has been made in elucidating the pathogenesis of IBD. In clinical practice, it is considered that the complex interaction between genetic susceptibility, environmental factors and intestinal mucosal barrier leads to abnormal oxidative stress, autophagy and mucosal immune response. This paper briefly summarizes and expounds the research progress of genetic and environmental factors, intestinal mucosal barrier function, oxidative stress, autophagy dysfunction and adaptive immune response, so as to provide basis for further research and development of treatment methods and improvement of clinical treatment effect.
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Effect of Bushen Jianpi Kaixin formula on autophagy and apoptosis in AD rats
li ying, Wang Ying, Kong Mingwang
Abstract:
Objective To investigate the effects of Bushen Jianpi Kaixin Formula (BSJPKXF) on the learning and memory ability of Alzheimer s disease (AD) model rats and the related autophagy and apoptosis in cortex of AD model rats, and exploring the underlying mechanism of BSJPKXS. Method The 60 eligible rats were randomly divided into six groups (n = 10): control group, AD group, Bushen group (BS, 3.6g/kg·d), Jianpi group (JP, 4.05g/kg·d), Kaixin group (KX, 2.34g/kg·d) and Bushen Jianpi Kaixin group (BSJPKX, 9.99g/kg·d). AD mouse model was established by intraperitoneal injection of D-gal. The rats in BS group, JP group, KX group and BSJPKX group were gavaged with corresponding drugs once a day. The rats in control group and AD group were treated with equal amount of normal saline one time per day. After four weeks, the learning and memory ability was tested by Morris water maze. The open-field test was used for detecting the cognitive function in rats. The expression of LC3-I, LC3-II and Beclin in cerebral cortical tissues were detected by western bolt. The expression of Bax and Bcl-2 in cerebral cortical tissues were detected by immunohistochemistry. The related mRNA level of Beclin 1, P62, Bax and Bcl-2 in cerebral cortical tissues were detected by RT-PCR. Result Compared with the control group, D-gal significantly decreased the spatial learning and memory ability of rats in the AD group (P < 0.01), decreased the expression of Beclin, LC3-I/LC3-II, Bcl-2 and Bcl-2/Bax, increased the mRNA level of P62 and the expression of Bax (P < 0.01). After treatment, related to the AD model group, Bushen Jianpi Kaixin formula improved the spatial learning memory ability of rats in the BSJPKX group (P < 0.01), increased the expression of Beclin, LC3-I/LC3-II, Bcl-2 and Bcl-2/Bax, decreased the mRNA level of P62 and the expression of Bax (P < 0.01). Conclusion Bushen Jianpi Kaixin formula can improve cognitive impairment in AD rats. The mechanism is presumedly related to the reduction of neural autophagy and apoptosis.
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The evaluation of experimental animal studies on acupuncture for hypertension based on the SYRCLE’s tool and the ARRIVE guidelines
Tang wenjing, ZHANG Yue, LI Yuxi, ZHONG Dongling, JIN Rongjiang, ZHANG Lili, LI Juan
Abstract:
Objective To evaluate the risk of bias and reporting quality of animal experimental studies on acupuncture for hypertension, and analyze the deficiencies in experimental design, implementation and reporting, so as to decrease the risk of bias of researches and improve the reporting quality of animal studies. Methods Databases including China national knowledge infrastructure（CNKI）, Wanfang database（WANFANG）, Chinese science and technology periodical database（VIP）, Chinese biomedical literature database（CBM）, Web of Science, PubMed, Embase, and The Cochrane Library were searched from inception to October 8, 2022. Two researchers independently screened experimental studies related to acupuncture interventions in hypertensive animals according to eligibility criteria. The SYRCLE’s tool and the ARRIVE guidelines 2.0 were used to evaluate the risk of bias and reporting quality of the included literature. Excel 2019 was used to extract data and descriptive analysis was conducted according to the results. Results A total of 79 animal experiments on acupuncture for hypertension were included, among which 16 and 21 were published in Chinese core journals and science citation index（SCI）journals respectively. The result of SYRCLE’s tool showed that 5 of the 10 items were evaluated as well, while the rest were unclear risk or high risk of bias. Of the 79 included articles, the risk of bias of 19 articles were low, while the remaining studies had some risk of bias. The results of the ARRIVE guidelines 2.0 indicated that 19 of the 38 sub-items were well reported, while the rest were poorly reported.The reporting quality of 51 articles were good and the remaining studies were inadequately reported. Conclusions At present, the risk of bias of acupuncture intervention in hypertensive animal experiments is high and the reporting quality is generally low. The inadequate description of some important entries had impact on the reproducibility of the experiments and the translation of the results.The SYRCLE’s tool and ARRIVE guidelines 2.0 are recommended to be used as reference during experimental design and reporting, thus improving the standardization and reporting quality of acupuncture for hypertensive animals.
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Research advances of trophoblast cells in threatened abortion complicated with intrauterine hemorrhage
HuoYan, Zhu Jun nan, Fu Ya qi, Chen Ping, Xu Guang li
Abstract:
Threatened abortion complicated with intrauterine hemorrhage is a common disease during pregnancy.Its pathogenesis is related to the imbalance of maternal-fetal interface microenvironment and the obstruction of uterine spiral artery remodeling. Trophoblast cells are the outermost layer of the maternal-fetal interface microenvironment .Its proliferation, migration and invasion are closely related to normal pregnancy.It plays an irreplaceable role in immune tolerance and regulation, uterine spiral artery remodeling, and maintenance of maternal-fetal interface microenvironment.In recent years, trophoblast cells have become the main line of research in autoimmune diseases and other fields.Studies have found that it plays an important role in the diagnosis and treatment of threatened abortion complicated with intrauterine hemorrhage.This article reviews the research progress of trophoblast in the treatment of threatened abortion complicated with intrauterine hemorrhage.
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ZHANG Gedi, LIU Gengxin, LUO Fuli, YAN Ziyou
2023,33(7):17-25, DOI: 10. 3969 / j.issn.1671-7856. 2023. 07. 003
Abstract:
Objective To predict the mechanism of Shenshuai Prescription (SSR) in chronic kidney disease(CKD)-related myocardial injury using network pharmacology and molecular docking method. Methods We used the traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP) and Herb herbal medicine identification database (http:/ / herb. ac. cn/ ), and the SwissTargetPrediction database to screen target information of active ingredients. We then used the UniProt database to screen for human targets and standard gene names. A drug active ingredient target network diagram was constructed using Cytoscape 3. 7. 2 software, and the GeneCards database was used to collect disease-related targets. The “ Shenshuai Recipe” against CKD myocardial injury gene target database was established using Venny 2. 1, and the STRING database was used to build the main component target interaction network and screen key targets. Cytoscape 3. 7. 2 software was imported for topology analysis and a protein-protein interaction network diagram was constructed. Finally, the DAVID platform was used for Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis and Gene Ontology (GO) biological function annotation. Results After screening, there were 252 active compounds in SSR and 649 common targets of SSR and CKD myocardial injury, among which AKT Serine/ Threonine Kinase 1 (AKT1), Tumor Necrosis Factor (TNF), Mitogen-Activated Protein Kinase 3 (MAPK3), and Vascular Endothelial Growth Factor A (VEGFA) may be important targets of SSR in treating CKD myocardial injury. GO analysis identified 1485 Biological process items, 176 Cell component items, and 386 Molecular function items, of which plasma membrane, cytosol, and cytoplasm had the largest number of enriched genes, and 313, 304, 276 genes were distributed respectively. KEGG analysis indicated that HIF-1α, Lipid and atherosclerosis, AGE-RAGE signaling path in diagnostic complexes, phosphoinositide 3-kinase (PI3K)-AKT, and insulin resistance pathways might be involved in the mechanisms of SSR in treating CKD myocardial injury. Conclusions SSR might play a role in cardiorenal protection by participating in multiple mechanisms, including improving insulin resistance, improving lipid metabolism, antiatherosclerosis, and regulating the expression of inflammatory factors and vascular endothelial growth factor, with the PI3K/Akt and mitogen-activated protein kinase pathways being potentially important signal regulation pathways.
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WANG Xue-qiao, RAO Yu-teng, WU Wei-peng, CHENG Peng, LI Wei, JIANG Dai-xun, CHEN Wu
2014,24(6):22-26, DOI: 10.3969.j.issn.1671.7856.2014.006.005
Abstract:
Objective To establish a canine model of intervertebral disc extrusion by surgery and observe the histological and microcirculatory changes of the spinal cord, in order to accumulate data for studies on the pathology and mechanism of treatment for intervertebral disc extrusion. Methods Normal healthy adult dogs were divided randomly into two groups: normal control group and model group. To simulate the intervertebral disc extrusion caused by spinal cord compression, 6Fr double lumen catheter with ballon was inserted into the spinal cord T12-T13 and filled with about 5 mL Iohexol after the exposure of spinal cord L1 by hemilaminectomy. The spinal cord blood flow (SCBF) at the L1 level before and after compression was measured by laser-Doppler flowmetry. Morphological changes of the compressed spinal cord at 14 days after compression was examined by histopathology.Results The (Texas spinal cord injury score) (TSCIS) scores of the motor function of bilateral hind limbs were highly significantly decreased (P<0.01). The blood flow of spinal cord at the L1 level was significantly decreased (P<0.05) after compression than that before. Compared with the normal control group, the model group showed abnormal vacuolization in the white matter and the number of normal neurons in the ventral horn of gray matter was significantly lower(P<0.01). Conclusions Our findings demonstrate that canine models of intervertebral disc extrusion can be successfully established by balloon catheter compression, showing local impairment of microcirculation and histological changes in the spinal cord. This canine model may provide a useful model for evaluation of therapeutic effects of acupuncture and for mechanism studies.
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2017,27(5):19-22, DOI: 10.3969.j.issn.1671-7856.2017.05.007
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In this paper, the present situation and development trend of the laboratory animal industry in China are expounded comprehensively and objectively. Through comparative analysis, relative to Europe and the United States one hundred years of accumulated huge resources and strong financial support, the overall laboratory animal industry in China is far behind. The whole nation system should make great efforts to realize leaping development and to catch up. Laboratory animal resources are still the foundation and weak link of the development of laboratory animal industry. This study can provide a scientific basis for the competent authorities at all levels to develop fast and efficient laboratory animal industry development strategy, and also provide reference for the development of the laboratory animal industry itself.
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LI Yan-hong, LIU Ying, QIN Chuan
2012(6):65-71, DOI: 10.3969/j.issn.1671.7856.2012.06.015
Abstract:
There are some correlations between the variation of thymus,T、B cells of adaptive immune system, neutrophil, macrophage, NK/NKT cells, Dendritic cells etc of natural immune system and immunosenescence. Immunosenescence mainly associated with the varies of adaptive immune system, therefor, this article will introduce the inordinately varies of the number, surface molecules, cytokines and signal transduction as well as the dysfunction of T and B cell with age.
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CHEN Ting, LI Feng-di, DENG Wei, QIN Chuan
2014,24(1):72-73, DOI: 10.3969.j.issn.1671.7856.2014.001.017
Abstract:
Objective To compare mouse inner canthus veniplex injection and tail intravenous injection.Methods 4～6 weeks BALB/c mice were injected with normal saline respectively, the time and the difficulty were compared. Results mouse inner canthus veniplex injection is easier than tail intravenous injection.Conclusion mouse inner canthus veniplex injection is an easier and more successful method for mouse intravenous injection.
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ZHAO Xing-mei, WANG Yan, CHEN Hua-de
2014,24(2):38-41, DOI: 10.3969.j.issn.1671.7856.2014.002.009
Abstract:
Objective To study the effects of Shenqi-fuzheng injection (SQ) in different doses injected into Zusanli acupoint in treating heart-qi deficiency syndrome and explore its dose-to-effet relationship.Methods The rat model of heart-qi deficiency syndrome was established by forced burden swimming and gavage of large doses of propranlolum. Seven groups of rats were set up with 8 rats in each group: the blank control group, the model group and 5 treatment groups which received a Chinese medicine Shenqi-fuzheng injection (SQ) in different doses (0.05 mL, 0.10 mL, 0.15 mL, 0.20 mL and 0.25 mL per rat, respectively) at ST36 acupoints. The treatment was administered for consecutive 10 days. The general conditions and symptoms of the rats were observed and recorded. The serum concentrations of atrial natriuretic peptide (ANP) and cyclic adenosine monophosphate (cAMP) were assayed by ELISA. The activity of serum superoxide dismutase (SOD) was detected by colorimetry. The pathological changes of myocardial tissue were observed using HE staining. Results Compared with the blank control group, the model group showed obvious heart failure symptoms such as fatigue, purple tongue, short and rapid breath, the concentration of ANP was increased while cAMP decreased, and SOD activity was declined (P<0.001 for all). The pathology of myocardial tissue showed inflammatory cell infiltration and seriously swollen cardiomyocytes in disordered arrangement. Compared with the model group, all treatment groups showed that symptoms were relieved, ANP concentration was decreased, and cAMP concentration and SOD activity were increased. Among them, the changes of indicators in the SQ-0.05 mL group were mildest (P<0.05) while strongest in the SQ-0.20 mL group (P<0.001). The pathological changes of myocardial tissue in the SQ-0.20 mL group were mildest and close to normal. Conclusions ST36 acupoint injection with the Chinese medicine Shenqi-fuzheng injection is an effective therapy for heart-qi deficiency syndrome in rats. The treatment effects are positively correlated with the doses of drug in the range of from 0.05 mL to 0.20 mL per rat.
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ZHAO Yong, WU Jing, MAO Feng-feng, ZHAO Shan-min, ZHANG Cai-qin, BAI Bing, SHI Chang-hong
2012(7):17-20, DOI: 10.3969.j.issn.1671.7856.2012.007.005
Abstract:
ObjectiveDiscussion the common problem of tumor-burdened in node-mice,In order to improve the success rate of tumor burden. MethodThrough analysis the influence of various factors of tumor-burden in node-mice, seek the solution of problem in tumor burdended. ResultSummarized the various factors in the tumor burden, and put forward reasonable suggestions.ConclusionNode-mice tumor burden is the base of the oncology, drug and biological products of safety evaluation and effectiveness screening. This paper is to provide reasonable suggestions for the preparation of good nude mice tumor model.
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2013,23(11):0-0, DOI:
Abstract:
[Abstract] Objective To study the effects of D-galactose on the expression of P-ERK 1/2 in hippocampus of mice and on learning and memory ability Methods Twenty Kunming mice, half male and half female, were divided into two groups. To establish aging model of mice, D-galactose was injected daily in the subcutaneous for 35 days in the dosage of 125 mg / kg. The control group was given saline. In the modeling period, the general condition of mice was observed. Step-down and morris water maze were carried out to test the learning and memory ability at the last one week. After the experiment, the animals were sacrificed to extract hippocampal tissue. The expression of P-ERK1 / 2 in hippocampal of mice was detected by western blot. Results Step-down test showed that compared with the control group, the latency in the model group decrease significantly (P <0.01) and the number of electric shock in the model group increase significantly (P <0.01). Morris water maze showed that the latency of escape in the model group were longer than the control group (P <0.01). Western blotting results showed that the expression of P-ERK1 / 2 significantly decreased in the model group and the effect is more pronounced in male mice. Conclusion D-galactose may impair learning and memory ability by inhibiting the expression of P-ERK1 / 2 in hippocampal of mice and the effect is more pronounced in male mice.
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2013,23(11):0-0, DOI:
Abstract:
Objective To analyze the difference of growth and organ coefficient between mutant mice and normal mice using inbred BALB/c curly mutant mouse model．Methods 21d，42d and 63d three different age groups of BALB/c mutant curly and normal BALB/c mice of 20 each，half male and half female，were selected to measure body weight，head length，body length，tail length and to compare the head body ratio，tail body ratio and organ coefficient．Results F4 generation BALB/c mutant curly has completely homozygous mice．Head body ratio，tail body ratio of mutant curly mice aged 21d and 42d were lower than those of normal mice (P< 0.05)，body weight，head body ratio，tail body ratio of mutant curly mice aged 63d were lower than those of normal mice (P< 0.05)．Heart，spleen，ovarian，uterine coefficient of 21d mutant mice were greater than those of normal mice (P< 0.05，P＜0.01)；heart and thymus coefficient of 42d mutation curly mice were lower than those of normal mice，brain and testis coefficient were higher than those of normal mice (P< 0.05，P< 0.01)；heart and uterus coefficient of 63d mutant curly mice lower than those of normal mice，brain coefficient was higher than those of normal mice (P<0.05，P< 0.01)．Conclusion There are differences in appearance and organ coefficient of BALB/c mutant curly and normal mice，these different mutations have very important significance for the study of BALB/c mutant curly mice growth，cardiovascular system，immune system and reproductive system．
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WANG Jun-feng, SHI Mei-lian, WU Yan-qiu, GUO Jian-guang, FU Jie, SUN Zong-guo, GAO Jing-hua, XU Ping
2012(3):17-22, DOI: 10.3969/j.issn.1671.7856.2012.03.004
Abstract:
ObjectiveTo determine the growth rate, organ index, blood Physiological and biochemical Parameters of germ-free KM mice during different growing Period, baby germ-free KM mice were obtained by hysterectomy form SPF KM mice and reared by hand feeding under aseptic condition. Methods(1) Weight and body size of the mice were measured at different days(from 0 to 112days; n=60; 30♀,30♂) to create a growth curve (2) The weight of the live mice and major organ size after dissection were measured at different days selected randomly (day = 8,6,112; n = 60 30♀,30♂) to determine the growth of these mice. (3) The blood Physiological and biochemical Parameters were measured by collected blood samples from the infraorbital vessels. Results(1) The weight of germ-free KM mice increased with the growing days, and the growth rate reached maximum within 1-2weeks Post-weaning. Significant difference was found between males and females(P＜0.05) after 4th week. (2) All the major organs (heart, liver, spleen, lung and brain) increased with the ages except the thyme gland, and the growth of cecum size was obvious. (3) Difference of blood Physiological and biochemical Parameters was found between males and females at the same age, and the difference types increased with the growth of mice age. ConclusionThe results of growth rate and blood physiological and biochemical parameters could provide reference for the evaluation of germ-free KM mice colony.
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ZHAO Jing-shu, WANG Rong, ZHANG Jing-yan, WU Yan-chuan, JI Zhi-juan
2012(2):66-69, DOI: 10.3969/j.issn.1671.7856.2012.02.015
Abstract:
ObjectiveTo establish an in vitro model of caloric restriction (CR) with human neuroblastoma cell line SH-SY5Y cells and observe the effects of different energy supplies on the growth of SH-SY5Y cells. MethodsHuman neuroblastoma cell line SH-SY5Y cells were cultured in different media with glucose at different concentrations:low glucose group (2 g/L), normal glucose group (3.15 g/L) and high glucose group (4.5 g/L). MTT metabolic rate, cell growth curve and LDH leakage rate were measured to observe the cell growth status of different groups. ResultsCompared with the normal control group, the cells of the high glucose group had shorter cytoplasmic processes and shrank cell bodies, showed a little bit lower MTT metabolic rate, higher LDH leakage rate and poorer growth status, while the cultured cells of the low glucose group had more extensive processes, greatly lower MTT metabolic rate and lower LDH leakage rate, and the cells grew slower but showed good morphology. ConclusionsCulture at high glucose concentration is injurious to cells and induces a higher metabolic rate, so the cells are inclined to be senescent and to die. Culture at low glucose concentration in an allowable range of caloric restriction is protective to the cells, unharmful to the cells, and favorable for cell growth and prolongs the cell survival.
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LIU Qin, WANG Li-ping, CHEN Fang, ZHANG Yi
2015,25(7):69-73, DOI: 10.3969.j.issn.1671.7856. 2015.007.015
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Adipose-derived stem cells(ASCs) as potential seeded cells have been widely used in tissue engineering. Thus to obtain enough, high activity, high purity adipose-derived stem cells is the particular important premise of the application in tissue engineering. In this paper, the isolation and purification methods of ASCs were reviewed and the merit and demerit of different methods were compared in order to provide theoretical basis for safe and high-effective isolation and purification of ASCs.
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ZHANG Zhicheng, YUAN Yuan, WANG Xuan, SONG Qingkai, DAI Jiejie
2018,28(5):21-27, DOI: 10.3969/j. issn. 1671 -7856. 2018. 05. 005
Abstract:
Parkinson’s disease (PD) is a progressive neurodegenerative disorder, with an etiology that is now considered to be due to interaction between genetic and environmental factors. Typical PD features include loss of dopaminergic neurons in the nigrostriatal region, with typical motor traits of PD associated with dopamine deficiency. Animal models have contributed to determining PD etiology and pathogenesis, as well as testing new therapeutic schedules and novel drug research. Rodents, tree shrews, primates, and other animal models of PD have been established by different method . These models each have their own advantages and limitations, showing different clinical features and pathological mechanisms to those in humans. Therefore, the appropriate model for scientific research must be carefully considered. This article reviews the main neurotoxic and transgenic models of PD.
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LOU Qi, SHI Qiao-juan, GUO Hong-gang, LI Wei, LU Ling-qun, ZHOU Wen-wei, SA Xiao-ying
2012(3):5-11, DOI: 10.3969/j.issn.1671.7856.2012.03.002
Abstract:
ObjectiveTo investigate the pathogenesis of nonalcoholic fatty liver disease(NAFLD), provide a reference for the evaluation of fatty liver therapeutic effect, the liver function, lipid metabolism,insulin resistance and liver apoptosis of NAFLD rats established by high cholesterol feeding were continuously monitored from 4~16 weeks. Methods 40 NAFLD rats established by high cholesterol feeding were randomly divided into the 4,8,2,6 weeks model group and a group of normal rats were set up for control, the serum GLU、CHO、TG、HDL、LDL、GPT、GOT and insulin were tested, the liver apoptosis and pathological changes were observed, then analyzed the effect of the liver function, lipid metabolism,insulin resistance and apoptosis on the pathological changes in non-alcoholic fatty liver rats. ResultsAfter 4 weeks of high cholesterol feeding, the rats had got the liver injury, disorder of lipid metabolism, insulin resistance. Liver cell apoptosis were increased obviously, and pathological changes were characterized by cell steatosis and inflammatory, the lesions became more serious with time went on. ConclusionsRats established by high cholesterol feeding could get the nonalcoholic fatty liver disease and the lesions became more serious with time over during 4~16 weeks. Lipid metabolism disorder and insulin resistance were the important cause of liver cell steatosis and inflammatory. The model we established is suitable for the evaluation of fatty liver therapeutic effect.
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PAN Yongming, CHEN Liang, Xu Jian-qin, Zhu Ke-yan, Chen Min-li
2013,23(11):0-0, DOI:
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[Abstract] Objective To establish the disease syndrome combined animal model, the WHBE rabbit myocardial ischemia model of phlegm-blood stasis syndrome type, by high fat diet feeding and coronary artery ligation. Methods WHBE rabbits were randomly divided into the normal control (NC) group, myocardial ischemia model (MI) group and myocardial ischemia model of phlegm-blood stastis syndrome (PBSS-MI) group, 6 in each group. The myocardial ischemia of MI group and PBSS-MI group were induced by ligated the coronary artery, and then NC group and MI group were fed with common forage, while PBSS-MI group was fed with high fat forage for 4 weeks. The model establishment and its pathological process of phlegm-blood stasis syndrome were evaluated by examinations on body weight, myocardial enzymes indexes, blood liquid levels, hemorheology and myocardial infarction size (MIS), and correlation analysis by Pearson method. Results Compared with NC, the weight of MI group was slightly lowered, and the serum CK, LDH, TG, plasma viscosity, casson viscosity and myocardial infarction size were significantly increased (P<0.05, P<0.01).While the weight of PBSS-MI group had no obviously lowered, but serum CK, LDH, TC,TG, LDL-C, HDL-C, whole blood viscosity, plasma viscosity, erythrocyte electrophoresis time, casson viscosity and myocardial infarction size were significantly increased (P<0.05, P<0.01). Compared with MI group, TC, LDL-C, HDL-C, high shear rate of whole blood viscosity, erythrocyte electrophoresis time and myocardial infarction size were higher than those of MI group (P<0.05, P<0.01). Correlation analysis showed that MIS and TC, TG and HDL-C in MI group were closely related (P<0.01), and MIS and TC,TG, HDL-C, LDL-C, LDH, CK, high shear rate of whole blood viscosity, low shear rate of whole blood viscosity, erythrocyte electrophoresis time and cassion viscosity in PBSS-MI group were also closely related (P<0.05, P<0.01). Conclusion Disease syndrome combined model for myocardial ischemia model of phlegm-blood stasis syndrome type in WHBE rabbit could be established by high fat diet feeding with coronary artery ligation, and it has related changes between MIS and blood lipids level, hemorheology, which may reflect related biological basis of the traditional Chinese medicine of phlegm and stasis cementation.
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LI Bi-hai, LUO Li-juan, LIANG Xin-lin, JIN Shi-jun, RAO Jun-hua
2014,24(1):23-25, DOI: 10.3969.j.issn.1671.7856.2014.001.006
Abstract:
Objective To explore the correlation between vaginal bleeding during early pregnancy and abortion of Macaca fascicularis. Method The Statistic results of the incidence of early pregnancy bleeding and abortion rate were worked out by observing the normal menstrual and the vaginal bleeding during early pregnancy. Then the correlation between vaginal bleeding during early pregnancy and abortion of was investigated based on the results. Results The ratio of Macaca fascicularis bleeding in the first menstrual cycle was reached 80% of the total amount of pregnancy, while the abortion rate was 0%. Conclusion Since the vaginal bleeding in early pregnancy was one basic phenomenon of Macaca fascicularis, it was have no effect or little impact on pregnancy security.
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LI Rifei, YUAN Na, YE Dongyang, LI Riyong, LI Yaohan, ZUO Runan, SHI Xi, CHEN Qiang, LI Yinqian
2018,28(2):113-118, DOI: 10.3969. j. issn. 1671 -7856. 2018. 02. 020
Abstract:
Breast cancer is the most common spontaneous malignancy in women, causing a serious threat to women’s health. Experimental animal models have been important research tools in bringing insights into the related mechanisms of breast cancer, as well as in delivering improved therapies for the disease. To reveal the pathogenesis of various breast cancers and facilitate the development of new drugs, ideal animal models of breast cancer should share some common tumor molecular characteristics and biological behavior with that of human breast cancer. In this review, five types of experimental animal breast cancer models were included, which may provide a basis for research on diagnosis and treatment of human breast cancer.
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ZHANG Lei-yang, HE Min, LI Yue, QIU Fu-rong, CHEN Wen-wen, WU Yu, YANG Jiang-wei, JIANG Jian
2017,27(9):92-97, DOI: 10.3969.j.issn.1671-7856.2017.09.018
Abstract:
The incidence of depression has been increasing over the recent years, which can cause serious physical and mental health problems in humans, but its pathogenesis has not been fully clarified. Animal models can simulate the depression in humans, thus are widely used for studies of the pathogenesis of depression, as well as in research and development of new antidepressants. According to the different ways of modeling, animal models of depression can be divided into the following categories:stress models, surgical models, drug-induced models and genetic models. These models can provide useful tools to explain some pathogenetic aspects of depression, such as neurotransmitters and their receptors/transporters, neurotrophic factors, neuroendocrine systems, inflammatory hypotheses, and so on. This review summarizes and evaluates the commonly used rodent animal models of depression and provides a reference for further research on depression.
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GE Wen-ping, ZHANG Xu, GAO Xiang, LIU Ning, ZHANG Hai-tao, ZHANG Xiao-juan
2012(3):65-68, DOI: 10.3969/j.issn.1671.7856.2012.003.014
Abstract:
ObjectiveTo provide reference for producing a good laboratory animal production and management, we randomly analyzed microbes and parasites of Specific pathogen free(SPF)mice commonly used in China.Methods The current national standards《Laboratory animal—microbiological standards and monitoring》and《Laboratory animal—parasitic standards and monitoring》were used to tested the microbes and parasites in the samples of SPF mice strains commenly used in China, including ICR,KM,C57BL,BALB/c and BALB/c-nu.ResultsThe results showed that the main virus carried by SPF mice were Reo-3, PVM and POLY, the bacteria contamination included klebsiella pneumoniae, Staphylococcus aureus and Peudomonas aeruginosa. They all affect the mice and experiment in certain degree except for Peudomonas aeruginosa.
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2013,23(9):0-0, DOI:
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[Abstract] Objective: To study the toxicity of intravenous injection of human umbilical cord mesenchymal stem cells in C57/BL6J mice with repeated administrations. Methods: 32 SPF C57/BL6J mice were randomly divided into negative control group and HUMSC transplantation group, of equal gender. HUMSCs were isolated and subsequently cultured through 5 passages in vitro. 2?06 HUMSCs were injected each mice of transplantation group via tail vein once 1 week for 4 weeks, and the equal quantity PBS were injected to control group mice. Mice were euthanatized on the day of 1 week (8 mice per group) and 4 weeks(all survived mice) after the last treatment following the schedule. Toxicity was evaluated by clinical observations, pathology (blood cell counts, clinical biochemistry), immunologic consequences, visceral organs weight and anatomic pathology. Results:No significant difference in hematological and biochemical parameters between HUMSC transplantation group and control group (P > 0.05). There was also no significant change in organ weight or organ coefficient between the two groups(P > 0.05). There was no significant change in the subpopulation of T-cells (the percentage of CD3, CD4, CD8 T cells, and the ratio of CD4 and CD8 T cells). HUMSC transplantation group did not show any histopathological change in the viscera organs compared to control group. Conclusion:Results have suggested that intravenous injection of HUMSCs in C57/BL6J mice with repeated administrations is safe and feasible. No side effects and immune response were found in recipients.
