• Issue 3,2005 Table of Contents
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    • Pathological Changes in SARS-CoV Infected Rhesus Monkeys

      2005(3):132-135.

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      Abstract:目的在感染的8只恒河猴的SARS-CoV模型动物中,观察肺等组织中出现的系列病理学改变,为针对抗SARS药物筛选、疫苗评价中的免疫病理反应等奠定实验依据。方法SARS-CoV经鼻腔接种8只恒河猴,在感染的第5、7、10、15、20、30和60天,分别安乐处死动物,组织病理取材,制片,观察。结果经病毒分离和RT-PCR证实动物感染是成功的。系列病理改变表明,早期肺组织可见间质性肺炎,水肿、结构破坏、出血,巨噬细胞浸润;后期出现内皮细胞受损及再生,透明膜形成,小血管玻璃样变,肺组织纤维化及肺气肿形成,肺泡网状纤维和弹力纤维破坏并增生等,脾脏、淋巴结生发中心早期有萎缩,后期有恢复等病理学改变均和SARS患者相似。结论感染恒河猴出现与SARS患者类似的临床和病理学改变,为进一步研究该病毒的病原特性、发病机理、药物筛选、疫苗评价等方面的研究奠定了重要基础。

    • Exploration on SARS-Coronavirus Infection Combined with Histoplasma Capsulatum

      2005(3):136-139.

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      Abstract:Objective To explore the etiology of SARS that it could be coronavirus infection combined with Histoplasma Capsulatum. Methods Specimens taken from one case of SARS autopsy and one case of rhesus monkey necropsy. Histopathological and immunofluorescence techniques were used to detect the fungal Histoplasma Capsulatum and pathological changes in the specimens of human and monkey's lung, spleen, lymph nodes and liver. Results 1. The histopathological changes of monkey were very similar to those in human beings. 2. The results of immunohistochemistry demonstrated that: (a) SARS convalescent serum and rabbit anti-Histoplasma Capsulatum serum vs. H. Capsulatum antigen revealed to be positive. (b) Both two kinds of serum vs. specimens of SARS and monkey's lungs, spleen, lymph nodes and liver appeared H. Capsulatum positive. Conclusion It was evident that the patient and monkey were infected with H. Capsulatum in vivo. The etiology of SARS is coronavirus combined with Histoplasma Capsulatum infection.

    • Report on A Deep Fungus Found from the Serum of SARS Patients

      2005(3):140-143.

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      Abstract:目的从临床诊断的SARS病人血清中分离病原微生物。方法采用SARS患者血清通过NIH小鼠腹腔、脑和鸡胚卵黄囊和尿囊接种、人胚肺细胞呼吸道病毒培养和系列细菌培养分离病原体,所获微生物经形态学、血清学、生化学和法国梅里埃(Vitek)全自动微生物系统鉴定,复制动物模型和病理学检查。结果从SARS病人血清分离的微生物,检定结果为一种双相型深部嗜肺性真菌,属流行性肺组织胞浆菌(球状酵母菌)。结论分离的肺组织胞浆菌(球状酵母菌)是呼吸系统深部真菌。

    • Virologic and Serologic Detection of SARS-CoV in Infected Rhesus Monkeys

      2005(3):144-146.

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      Abstract:Objective To study virological and serological detection of SARS-CoV in the infected Rhesus monkeys. Methods Eight rhesus monkeys were infected with SARS-CoV inoculated into the nasal cavity, and euthanatized at 5, 7, 10, 15,20, 30 and 60 days after inoculation, respectively. Throat swabs and blood samples were collected periodically for detecting the SARS-CoV and antibodies by RT-PCR, virus isolation and ELISA. Results Infections in the monkeys have been successfully established by SARS-CoV inoculation. The SARS virus RNA was detected on 5-16 days after inoculation using nested RT-PCR in throat swab samples. Viruses were isolated in 5 out of 8 infected monkeys. The SARS coronavirus-specific IgG was positive from 15 th day post infection. Conclusion The duration of PCR detection, virus isolation and antibody production may be used as important criteria for anti-SARS drug studies and vaccine evaluation.

    • SARS-CoV Survey on Breeding Civet Cats in the Suburbs of Beijing

      2005(3):147-148.

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      Abstract:Objective To investigate the possible infections of SARS-CoV in civet cat bred in the suburbs of Beijing. Methods Pharyngeal swab, anal swab and fur samples collected from 30 civet cats were examined using RT-PCR for SARS-CoV. The neutralized antibodies against SARS-CoV from sera from the civet cats were checked. Results All samples from the civet cats were SARS-CoV negative, and no neutralized antibodies were positive. Conclusion There was no evidence to conclude that the civet cats inbred in the suburbs of Beijing were host of SARS-CoV.

    • Determination of Genetic Purity of Bombyx mori Inbred Strain by RAPD

      2005(3):149-153.

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      Abstract:Objective To analyze the genetic purity of silkworm (Bombyx mori) inbred strain IS-c108A for directing the breeding of silkworm as a laboratory animal. Methods 20 selected single primers were used to amplify randomly genomic DNA of 30 individuals in each batch of the inbred strain IS-c108A, parent strain c108 of this inbred strain and the control strain 871. The amplified fragment bands were analyzed to calculate the similarity index and the genetic distance among the individuals and batches. Results The frequency of polymorphic bands in the three batches of the inbred strain IS-c108A were 1.807%, 1.841% and 1.841%, respectively, and the average was 1.830%. The frequency of polymorphic bands was 7.207% between 30 individuals of the parent strain c108 of this inbred strain,while it was 7.08% between individuals of the control strain 871. The frequency of polymorphic bands between the inbred strain IS-c108A and c108 was 49.20%,while it was 58.33% between c108 and 871. Based on RAPD patterns, the calculated similarity index between 30 individuals in each batch of the inbred strain IS-c108A were 0.99581, 0.99555 and 0.99551, respectively, the average was 0.99562. Conclusion All the results implied that the inbred strain IS-c108A of silkworm has reached a high degree of genetic purity, and the silkworm has the advantage to obtain an inbred strain with high degree of purity easily.

    • A Rat Model of Type 2 Diabetes Mellitus Induced by High Fat Chow and Low Dose Streptozotocin Injection

      2005(3):154-158.

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      Abstract:目的观察高脂喂养联合低剂量STZ注射的SpragueDawley(SD)大鼠2型糖尿病模型的代谢特征、病理学以及胰岛分子生物学变化。方法4周龄雄性SD大鼠36只随机分为三组(1)正常对照组(Control)9只,普通饲料喂养。(2)高脂组(HighFatchow,HE)9只,高脂饲料喂养,为普通饲料中添加20%脂肪(猪油和蛋黄粉各50%)和20%蔗糖。(3)糖尿病组(DM)18只。喂养4周后腹腔注射STZ(40mg/kg)。所有大鼠做灌胃葡萄糖耐量(OGTT)试验。放免法测定血清胰岛素,免疫组化染色观察胰岛β细胞的形态学特点,彩色图像分析系统测定胰岛素表达量,RT-PCR测定胰腺β细胞胰岛素mRNA表达水平。结果糖尿病大鼠空腹血糖(FBG)、胰岛素水平(FINS)显著高于Control组和HE组大鼠(P<0.01),空腹血清甘油三酯(TG)和游离脂肪酸(FFA)水平显著高于Control组(P<0.05);胰岛β细胞吸光度(A)显著低于高脂组大鼠(P<0.05),降低11.6%。胰岛素免疫反应阳性区占胰岛百分比显著低于Control组和HE组,分别下降31.9%(P<0.05)和43.1%(P<0.01)。胰岛素mRNA表达水平显著低于HE组(P<0.05)。STZ注射后48h(基线值)大鼠FBG水平的分布情况为A组(FBG<10.0mmol/L)占7/18;B组(FBG10~19.9mmol/L)占5/18;C组(FBG≥20mmol/L)占6/18。STZ注射后9d的OGTT结果与基线值相比,B组OGTT值总体变化最小,A组FBG的变异最大,达到25%。结论高脂喂养联合低剂量STZ注射的糖尿病大鼠模型模拟2型糖尿病发生的主要病理生理过程,具有高血糖、高胰岛素血症以及血脂异常等基本特征。

    • Higher Efficiency of Transgenic Mouse Production Achieved by Double-pronuclear Microinjection

      2005(3):159-162.

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      Abstract:目的建立高效的转基因小鼠制备技术,为开展遗传工程动物模型研究奠定技术基础。方法通过向小鼠受精卵原核中注入不同浓度的DNA分子,筛选最适注射用DNA浓度;将K14/hCTLA4-Ig基因表达载体分子通过显微注射分别导入小鼠受精卵雌、雄原核,并设立单原核注射对照组;利用输卵管腹壶部穿刺移植法将注射后的小鼠受精卵移植于同期发情的受体母鼠;利用PCR对出生的转基因首建小鼠进行筛选。结果最适DNA分子浓度为10ng/μl;在单、双原核注射组胚胎2细胞卵裂率分别为52.3%(132/253)和45.0%(108/240),差异有显著性(P<0.05);注射胚胎移植后体内存活率分别为18.1%(24/132)和16.7%(18/108),差异无显著性;转基因首建小鼠阳性率分别为3/24和5/18,转基因阳性小鼠占总注射胚胎的比例为1.2%(3/253)和2.08%(5/240),差异有极显著性(P<0.01)。结论尽管双原核注射对胚胎的2细胞卵裂率有一定影响,但通过双原核注射可有效提高转基因小鼠的制备效率。

    • Establishment of STZ-Induced Diabetic Nephropathic Rat Model

      2005(3):163-165.

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      Abstract:Objective To establish a diabetic nephropathic rat model and to study it's kidney lesions. Methods 40 male Wistar rats were randomly divided into control and experimental diabetic groups, 20 per each group. Diabetic rats were induced by one intraperitoneal injection of streptozotocin (STZ) in a dose of 65 mg/kg. The rats were breed for 14 weeks. During the experiment, the blood glucose and urine glucose were measured and gross health status was observed. Finally, serum creatinine (SCr), blood urea nitrogen (BUN), urine protein and urine albumin were assayed, and renal pathological changes were examined by light and electron microscopy. Results The diabetic rats showed increased SCr, BUN, urine protein and urine albumin levels and hypertrophy of the kidneys. The renal pathology exhibited glomerular and tubulo-interstitial alterations. Conclusion The STZ-induced rat diabetic model have certain glomerulus and tubulo-interstitial lesions and can be used in diabetic nephropathy studies.

    • Inhibition of the Growth of Mouse Lewis Lung Cancer by VEGF Antisense Oligonucleotides

      2005(3):166-169.

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      Abstract:目的探讨血管内皮生长因子(VEGF)反义寡核苷酸对C57BL/6小鼠Lewis肺癌肿瘤生长的抑制作用。方法制备C57BL/6小鼠皮下肺癌模型30只,随机分为3组,每组10只VEGF反义寡核苷酸(ASPODN)治疗组、VEGF正义寡核苷酸(SPODN)治疗组及对照组。接种Lewis肺癌细胞后24h内,分别皮下注射ASPODN及SPODN进行治疗,对照组只注射生理盐水,每周2次,连续4周;观察各组小鼠肿瘤的生长情况、游标卡尺测量肿瘤体积大小。所有C57BL/6小鼠于接种后第25天先用二维超声观察肿瘤的实时图象,利用脉冲多普勒获取血流频谱,获得收缩期峰值速度(PS),阻力指数(RI)。断颈处死小鼠,光镜及电镜下观察肿瘤组织形态学改变及超微结构变化,免疫组化法检测微血管密度(MVD)。结果与对照组瘤重比较(7.83±0.78)g、VEGF-ASPODN组(4.49±0.43)g能明显抑制小鼠肿瘤生长(P<0.01)、VEGF-SPODN组(7.73±0.69)g则无明显作用(P>0.05)。VEGF-ASPODN组、VEGF-SPODN组抑瘤率分别为42.7%、5.9%。组织形态学及超微结构观察,VEGF-ASPODN对肿瘤生长具有抑制作用。VEGF-ASPODN组与对照组相比较PS及RI有明显差异(P<0.01);VEGF-SPODN组与对照组相比无明显差异(P>0.05)。结论肿瘤原位注射VEGF反义寡核苷酸能抑制小鼠肺癌生长。

    • Expression and Function of Apoptosis-related Genes bcl-2 and Bax in Stress-induced Gastric Ulcer in Rats

      2005(3):170-172.

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      Abstract:目的探讨凋亡相关基因bcl-2和Bax在应激性溃疡中表达的变化及中药组方应激宁对其变化的影响。方法Wistar大鼠水浸-束缚应激(WRS)4h,用免疫组织化学方法检测胃黏膜组织bcl-2和Bax蛋白的表达。结果bcl-2和Bax在正常大鼠胃黏膜组织中有散在的表达;应激宁组bcl-2阳性细胞数目较应激组增多,两者具有明显的差异(P<0.05);应激组Bax阳性细胞数目较应激宁组增多,两者也具有明显的差异(P<0.05);在应激组和应激宁组中,Bax阳性细胞数目均比bcl-2增加(P<0.05)。结论WRS后胃黏膜组织bcl-2和Bax的表达差异很可能参与了胃黏膜损伤和修复过程,应激宁对胃黏膜的损伤具有一定的保护作用。

    • RAPD Characterization of 5 Clonal Cell Strains from S180

      2005(3):173-175.

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      Abstract:Objective To set up differential specific maps of 5 clonal cell strains from S180 with RAPD. Methods The Genomic DNA of the 5 clonal strains from S180 including S2D9, S2D7, S1F11, S1H10 and S1B11 was amplified with RAPD-PCR using 23 single primers. The PCR results were assessed by electrophoresis on agarose gel. Results 3 of the 23 single primers were found to be different between amplified products from different clonal strains.Conclusion RAPD characteristics of the 5 clonal cell strains from S180 are apparently different.

    • Effects of Vitamin C Polyphosphate on Growth, Lymphocyte Proliferation and Content of Serum Corticosterone in Mice

      2005(3):176-179.

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      Abstract:目的探讨维生素C多聚磷酸酯对小鼠生长、淋巴细胞转化及血清皮质酮含量的影响以及淋巴细胞转化和血清皮质酮含量在热应激条件下的变化。方法在4组小鼠的饲料中依次添加35%的维生素C多聚磷酸酯,添加剂量分别为0(对照组)、500(实验Ⅰ组)、2500(实验Ⅱ组)和5000mg/kg(实验Ⅲ组),在(21±1)℃下饲养4周,对照组和实验Ⅱ组各取一半小鼠经(35±1)℃热应激处理24h。用特定生长率的方法测定小鼠的生长,用噻唑蓝还原法测定淋巴细胞转化,用荧光法测定血清中皮质酮的含量。结果各组间ICR小鼠的生长、淋巴细胞转化无明显差异;实验Ⅱ组的血清皮质酮含量明显低于对照组,其他组间差异无显著性。热应激后,淋巴细胞转化无明显变化;血清皮质酮含量较应激前有显著上升,但实验Ⅱ组仍显著低于对照组。结论在饲料中添加维生素C多聚磷酸酯对小鼠的特定生长率和淋巴细胞转化均无显著影响,但会降低小鼠血清皮质酮的含量。

    • Kidney Gene Expression Changes of Human ApoE4-Transgenic Mice by cDNA Array Analysis

      2005(3):180-183.

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      Abstract:目的研究人apoE4转基因鼠肾脏的基因表达谱变化.方法分别提取人apoE4转基因鼠和正常C57BL/6J小鼠的肾脏总RNA,经逆转录合成cDNA探针后分别与鼠cDNA表达点阵杂交,再用ESTblot软件进行分析,并用Northern印迹证明基因表达的改变.结果人apoE4转基因鼠肾脏中有38个基因的mRNA表达升高,22个基因的mRNA表达降低.其中血浆谷胱甘肽过氧化物酶前体、视黄酸γ受体和白介素5受体等基因的表达明显增加.B-raf原癌基因、促红细胞生成素受体、整联蛋白α4的基因表达显著降低.Northern杂交证明转基因鼠肾脏的c-Jun基因表达升高.结论人apoE4转基因鼠肾脏的c-Jun、血浆谷胱甘肽过氧化物酶前体、白介素5受体等基因的表达增加;促红细胞生成素受体、整联蛋白α4等基因的表达减少.

    • The Effect of Ceramide on Lactate Dehydrogenase Metabolism of Primary Cultured Mice Cortical Neurons

      2005(3):184-186.

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      Abstract:Objective To explore whether natural ceramide has effect on lactate dehydrogenase(LDH) metabolism in primary cultured mice cortical neurons. Methods Ceramide at different concentrations of 50,100,200,500,1000,2000 nmol/L was given to the culture medium of primary cultured mice cortical neurons. The LDH concentration was assayed after 0 h,1 h,4 h,8 h,12 h,16 h,24 h,36 h respectively. Its metabolism rate and leakage rate were calculated respectively. Results Ceramide changed significantly the distribution of LDH inside and outside of the neurons. Exogenous ceramide stimulated LDH elevation in supernatant in a dose-dependent and time-dependent manner, but had little impact on the total LDH metabolism of neurons. Conclusion Ceramide elevates the leakage rate of LDH and has no effect on total LDH metabolism of neurons.

    • Application of Tree Shrew as an Animal Model in Biomedical Research

      2005(3):187-190.

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      Abstract:Objective Tree shrew (Tupaia belangeri) is a newly developed animal model in biomedical research. The animal can be successfully infected with many human pathogenic viruses, such as human hepatitis viruses A-D, Chikungunya virus, herpes virus, etc. Tree shrew has been quite widely used in research among others in neuroscience, reproductive biology, immunology and sociopsychology.

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