Abstract:目的探讨一氧化氮(nitric oxide,NO)对局灶性脑缺血再灌注所致神经细胞损伤的影响及影响机制。方法将SD大鼠随机分为假手术组(N组)、脑缺血再灌注组(MCAO组)、脑缺血再灌注加侧脑室微量注射20 mmol/L的L-Arg 5μL组(L-Arg组)及脑缺血再灌注加侧脑室微量注射20 mmol/L的L-NAME 5μl组(L-NAME组),作脑缺血30 min,再灌注12 h2、4 h和2 d,冰冻切片,相邻切片分别作焦油紫染色、NOS免疫组化、NOS mRNA原位杂交、TUNEL法原位检测凋亡细胞。结果N组NOS的活性弱阳性表达;MCAO组术后24 h NOS的表达明显增强,与各组比较,P<0.05;L-Arg组术后12 h小血管内皮细胞出现NOS的阳性高表达,术后24 h神经细胞NOS的阳性表达最高,与各组比较,P<0.05;L-NAME组各时间点NOS活性的表达为阴性或可疑阳性,与各组比较P<0.05,NOS的活性明显受到抑制。凋亡细胞的计数结果为N组26.3±4.2个,MCAO组62±4.2个,L-Arg组40.6±2.7个,L-NAME组78.3±3.3个,P<0.05。结论适量NO可有效降低细胞凋亡的发生,减轻脑缺血再灌注所致的神经细胞的损伤。

Abstract:Objective To investigate the effect of vitamin E on transcription activity of liver CYP450 1A1,CYP450 1A2,CYP450 1B1 genes in mice treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin(TCDD).Methods Forty-five mice were divided into 5 groups,and the levels of TCDD and vitamin E were given at a dose of 0 and 0,100 and 0,100 and 20,100 and 100,and 100 ng/kg/d and 500 mg/kg/d,respectively.After 4 weeks,livers were taken to analyze the transcription activity of CYP450 1A1,CYP450 1A2 and CYP450 1B1 genes by real-time PCR method.Reaults There was an increasing tendency of liver CYP450 1A1,CYP450 1A2 mRNA levels in the mice treated with TCDD compared with those in the control group,while CYP450 1B1 mRNA level was significantly lower than that of the control group.When vitamin E was supplemented at a dose of 20 mg/kg,there was a decreasing tendency of CYP450 1A1 mRNA levels in comparison with the group treated with TCDD.The CYP450 1A1 mRNA levels increased with the increase of vitamin E dosage,and significantly increased at vitamin E 500 mg/kg compared with those in the group treated with TCDD.The CYP450 1A2 mRNA level was decreased with increase of vitamin E dosage,and was significantly lower at vitamin E 500 mg/kg than that of the group treated with TCDD.When vitamin E was supplemented in a dose of 500 mg/kg,there was an increasing tendency of CYP450 1B1 mRNA level compared to that in the group treated with TCDD. Conclusion These results suggested that TCDD can affect or partly affect transcription activity of liver CYP450 1A1,CYP450 1A2 and CYP450 1B1 genes in mice,and vitamin E can alleviate the effect of TCDD to a certain extent.

Abstract:目的了解2型糖尿病模型GK大鼠生长曲线、主要脏器重量、糖代谢等生物学特性,评价GK大鼠葡萄糖刺激的胰岛素分泌能力。方法采用51只雄性GK大鼠及15只年龄性别匹配的Wistar大鼠作为研究对象。测定13周龄GK、Wistar大鼠空腹血糖、23周龄GK大鼠空腹及随机血糖。随访GK及Wistar大鼠生长曲线,34~46周龄期间血糖、糖化血红蛋白。46周龄时行腹腔葡萄糖耐量实验(IPGTT),计算相关参数评价β细胞葡萄糖刺激的胰岛素分泌能力;之后处死大鼠,脏器称重。比较GK及Wistar大鼠间上述各指标差异。结果13周龄GK大鼠空腹血糖4.74±0.41 mmol/L,对照Wistar大鼠1.85±0.44 mmol/L(P<0.001)。23周龄GK大鼠空腹血糖7.88±1.96mmol/L,随机血糖9.91±3.52~13.46±4.13 mmol/L。7~20及34~45周龄期间GK大鼠体重高于对照Wistar大鼠(P<0.05),46周龄时无显著性差异。34~45周龄期间GK大鼠空腹血糖、进食后血糖、HbA1c均高于对照Wistar大鼠(P<0.05)。IPGTT曲线下面积分析示GK大鼠胰岛素曲线下面积(AUCi)、葡萄糖曲线下面积(AUCg)高于对照Wistar大鼠,胰岛素与葡萄糖曲线下面积比值(AUCi/AUCg)低于对照Wistar大鼠,差异均有显著性(P<0.05)。GK大鼠肾脏重量高于对照Wistar大鼠(P<0.05),余主要脏器重量差异无显著性。结论GK大鼠空腹血糖、进食后血糖、HbA1c水平升高,葡萄糖刺激的胰岛素分泌能力(GSIS)减退,葡萄糖刺激后胰岛素分泌早期相消失,晚期相代偿性增加,具有2型糖尿病特点;体重、血糖等生物学特性稳定。

Abstract:目的表达犬细小病毒VP2蛋白(CPV VP2),用于犬细小病毒病的诊断、疫苗研制和VP2蛋白功能研究。方法采用PCR方法对CPV VP2基因进行扩增,将CPV VP2基因克隆到毕赤酵母(Pichia pastoris)分泌表达载体pPICZαA中,构建真核重组表达载体pPICZαA-VP2,将该重组质粒线性化后,转化毕赤酵母菌GS115中,在甲醇诱导下表达CPV VP2,SDS-PAGE和Western blotting鉴定表达蛋白。结果成功扩增了CPV VP2基因,构建了真核重组表达载体pPICZαA-VP2在毕赤酵母菌中表达出约64.35 kD蛋白。Western blotting鉴定表明,表达VP2蛋白与犬细小病毒阳性血清有反应性。结论在毕赤酵母中成功地表达了CPV VP2蛋白,能被犬细小病毒阳性血清识别。

Abstract:Objective To determine the values of physiological and biochemical parameters in BWEL-SPF chickens at age of 10,40 and 60 weeks.Methods The values of T,P,BT,CT,ESR,systolic pressure,diastolic pressure,RBC,HCT,MCV,RDW,HGB,MCH,MCHC,PLT,MPV,WBC,N,E,B,L,M,TP,Alb,AST,ALT,Glu and TG and CHO in BWEL-SPF chickens at age of 10,40 and 60 weeks,both male and female,were determined by corresponding measurement and t-test was made to analyze the results.Results There was a significant difference between CHO values in male and female chickens at age of 10 weeks(P<0.01),a very significant difference between TG,Ca,Na and Mg values in male and female chickens at age of 40 weeks(P<0.01),a significant difference between CHO values in male and female chickens at age of 40 weeks(P<0.05),a very significant difference between BT,B,M,ALT and Glu values in male and female chickens at age of 60 weeks(P<0.01),and a significant difference between CT and TG values in male and female chickens at age of 60 weeks(P<0.05).Conclusion The values of the physiological and biochemical parameters were different between male and female BWEL-SPF chickens at age of 10,40 and 60 weeks.

Abstract:Objective To study the effects of the selective inhibitor of PDE5 on mouse oocyte maturation in vitro.Methods Mouse cumulus-oocyte complexes(COCs) or denuded oocytes(DOs) were collected and cultured in the medium supplemented with the selective inhibitor of PDE5 and the rates of germinal vesical breakdown(GVBD) and first polar body(PB1) extrusion were observed at the end of the culture.In addition,a mouse follicle culture model was also used.Results In isolated oocytes,PDE5 inhibitor zaprinast prevented significantly both spontaneous oocyte meiotic maturation and FSH-induced HX-arrested meiotic maturation.In addition,treated the follicles with the type 5 inhibitor significantly inhibited the follicle-enclosed oocyte maturation induced by FSH.Conclusion Collectively,these results suggest that PDE5 induces mouse oocyte maturation.

Abstract:目的 探讨一氧化氮 (nitric oxide,NO) 对局灶性脑缺血再灌注所致神经细胞损伤的影响及影响机制.方法 将SD大鼠随机分为假手术组 (N组)、脑缺血再灌注组(MCA O组)、脑缺血再灌注加侧脑室微量注射20 mmol/L的L-Arg 5 μL组 (L-Arg组) 及脑缺血再灌注加侧脑室微量注射20 mmol/L的L-NAME 5 μl组 (L-NAME组),作脑缺血30 min,再灌注 12 h、24 h和2 d,冰冻切片,相邻切片分别作焦油紫染色、NOS免疫组化、NOS mRNA原位杂交、TUNEL法原位检测凋亡细胞.结果 N组NOS的活性弱阳性表达;MCAO组术后24 h NOS的表达明显增强,与各组比较,P＜0.05;L-Arg组术后12 h小血管内皮细胞出现NOS的阳性高表达,术后24 h神经细胞NOS的阳性表达最高,与各组比较,P＜0.05;L-NAME组各时间点NOS活性的表达为阴性或可疑阳性,与各组比较P＜0.05,NOS的活性明显受到抑制.凋亡细胞的计数结果为N组26.3±4.2个,MCAO组62±4.2个,L-Arg组40.6±2.7个,L-NAME组78.3±3.3个,P＜0.05.结论 适量NO可有效降低细胞凋亡的发生,减轻脑缺血再灌注所致的神经细胞的损伤.

Abstract:Objective To establish a new inbred strain and observe the growth, fecundity and some biological characteristics of DA rats under a stable raising condition. Methods 15 pairs of DA rats were raised to observe their fecundity and 25 pairs of DA rats were raised to examine their blood Indices. The rats were imported from production stock in Danmark. Results The inbred strain of DA rats was successfully established. DA rats provided a litter size of 5.5 rats in average, survival rate of 90.8 %, and the blood indices were close to those of Wistar rats. Conclutsion DA rats have been adapted to the conditions of our animal center and the introduced strain is successful. However,attention should be paid to that some biochemical indices and organ weight and organ cofficents are affected by sex.

Abstract:目的观察口服软骨复合物对类风湿性关节炎(rheumatoid arthritis,RA)的预防作用。方法口服软骨复合物两周后,再注射弗氏完全佐剂(Freund's complete adjuvant,FCA)诱发小鼠RA模型,通过观察小鼠关节炎分数的变化,测定小鼠各器官的指数,检测小鼠血清中一氧化氮(nitric oxide,NO)和肿瘤坏死因子-α(TNF-α)水平以及关节组织病理学的变化来评价口服软骨复合物对类风湿性关节炎的预防作用。结果软骨复合物能较有效的改善关节的肿胀度,并显著地提高小鼠的胸腺指数(P<0.01),显著地提高脾指数(P<0.05),明显降低NO和TNF-α水平;组织病理学显示给药组关节的变形破坏得到减轻。结论软骨复合物能有效地起到预防佐剂性关节炎的作用,然而其治疗作用还有待进一步研究。

Abstract:Objective To explore the methodology regarding culture,proliferation and purification of adipose-derived stem cells(ADSCs),and to study their biological characteristics. Methods Fat tissue was obtained from mesentery and subdermal tissues of rats of different lineages.The tissue samples were digested with collagenase to obtain adipose stromal cells.Cells were isolated and purified based on their different reaction to digestion.Immunohistochemistry was used to characterize and identify the harvested cells.Growth of cells was observed with an inverted contrast phase microscope.Results Despite different productivity,adipose stromal cells from different source tissues and different lineages showed identical biological characteristics.After 2-3 days of culture,floating cells became adherent and grew into clusters.After several passages,these cells separated and changed into spindle-shaped.The adherent cells could be further classified into two types.The first type of larger size contained profuse lipid droplets and highly positive by oil red O staining.The second type was of slimmer-shape,with little intracellular lipid and negative oil red O staining.After passages,remaining floating cells,albeit in a small number,continuesly differentiating into cells of the second type,while cells of first type gradually diminished.Assessed by immunohistochemistry,cells of the second type were CD49d( ) and CD106(-),which were characteristic phenotype of adipose stromal cells. Conclusion With modified duration of digestion,our method can be used to culture,proliferate,and purify adipose stromal cells,despite difference regarding productivity existing among different tissue sources and diverse lineages.

Abstract:目的为利用抑制消减杂交(SSH)法筛选旋毛虫抗肿瘤相关基因,建立理想的动物模型获取可靠的实验样本。方法Balb/c小鼠随机分为检测组(Tester)和驱动组(Driver),Tester为经口服感染旋毛虫250条、400条和550条组,Driver为不接种组,在感染后11 d,两组同时在皮下分别接种SP2/0细胞2×106个/只,检测组和驱动组小鼠荷瘤后,于20 d后处死,采集两组肿瘤组织和脾脏组织,用天平称量肿瘤块的重量,游标卡尺测量肿瘤块3个互相垂直直径进行体积比较;为了检测两组小鼠免疫情况,又采用流式细胞术检测体内T淋巴细胞的动态变化,以便评估所需样本的可靠性。结果将肿瘤组织与正常组织分离后,经统计分析比较两组肿瘤块重量和体积大小的差异均极为显著(P<0.01)通过检测T淋巴细胞亚类,FACS共检测1×105个细胞,分别得到脾细胞中CD3 、CD4 和CD8 T淋巴细胞的数量,感染组小鼠机体的CD3 、CD4 、CD8 和CD4 /CD8 显著增高,在接种不同剂量的旋毛虫后荷瘤,小鼠脾脏特异的T淋巴细胞亚类:CD3 差异显著(P<0.05);CD4 差异显著(P<0.05);CD8 差异显著(P<0.05);CD4 /CD8 差异显著(P<0.05)。通过以上指标的测定,我们认为所建立的动物模符合SSH的实验要求。结论建立的旋毛虫抗实体瘤动物模型,可用于旋毛虫抗肿瘤差异基因筛选的实验起始材料,为进一步研究旋毛虫抗肿瘤分子机制创造条件。

Abstract:目的建立具有高转移潜力食管癌细胞株并研究其生物学特征。方法将食管癌细胞系EC109细胞悬液异位移植到SCID小鼠胃壁,约3个月后或动物濒临死亡时处死,行病理学解剖,将肉眼可见的纵隔淋巴结转移瘤块接种于SCID鼠皮下扩增,然后取小鼠皮下瘤组织块进行细胞培养,得到性状稳定的细胞株NMC109后,用MTT法分析细胞生长曲线,Western bloting法检测与细胞分裂增殖能力密切相关的TopoIIα表达,酶谱法检测MMP-2和MMP-9的活性,划痕实验和Transwell体外移动实验检测细胞的移动能力。结果与母本细胞EC109相比,所获得的细胞株NMC109其增殖能力和TopoIIα表达明显增强,MMP-9的活性明显升高,移动能力明显增强。结论获得了具有高转移潜力的食管癌细胞株。

Abstract:目的通过低硒动物模型研究低硒及富硒蒜对雄性大鼠精子的影响。方法将30只大鼠分成3组:对照组(喂常规饲料)、低硒组(喂低硒饲料)、补硒组(喂低硒饲料加富硒蒜粉),喂养12周。实验结束时股动脉采血测血清硒(Se)含量、谷胱甘肽过氧化物酶(GSH-px)、超氧化物歧化酶(SOD)活性、及丙二醛(MDA)含量;处死动物后睾丸称重计算睾丸指数,测精子数量和精子活率。结果血清硒含量:3组大鼠分别为0.42±0.13、0.06±0.08、0.30±0.07 mg/L;精子数量:31.7±3.8、12.4±4.1、28.7±2.9×109;精子活率:78.35±3.23、44.63±4.40、74.45±5.62%;低硒组明显低于其他两组(P<0.01)。结论低硒能够造成大鼠精子数量减少、精子质量下降;富硒蒜具有拮抗氧化作用,能够改善低硒大鼠精子的数量及质量。

Abstract:Objective To establish a data-base of infant growth criteria in rhesus monkey conoly.Method Body weight,body length,tail length,forelimb and hindlimb in infants were measured from 100 days to one year old.Results With statistical analysis and drawing growth chart of the animals,the growth of male and female animals was almost the same.After one year the growth rate of males was higher than that of female.Conclusion The results of this study provide a criteria in growth measurement of infant rhesus monkeys,and are useful in continually improving their nutrition and breeding technology.

Abstract:Objective To explore the effect of calcitonin gene-related peptide(CGRP) and ET_1 on hypertension.Methods 42 rats with normal blood pressure were randomly divided into operation,sham operation and control groups.The level of CGRP and ET_1 were determined at 2,4,6,8 and 10 weeks after treatment,respectively.Results The blood pressure in rats of the operation group was significantly higher than that in the control group(P<0.01),but there was not significant difference between that of rats in the sham operation and normal control groups.CGRP of the operation group was significantly higher than that of control group(P<0.05).ET_1 of the operation group was significantly higher than that of control group.No significant difference was found between the ET_1 and CGRP in the sham operation group and control group.Conclusion CGRP has a protective effect on blood vessels in renal arterial hypertension,but ET_1 exerts a vasoconstrictive action.

Abstract:Objective To evaluate the benefits and safety of Tibetan drug Wangla as a food and as a drug.Methods Rats and mice were used in this experiment.Tests of acute oral toxicity,micronucleus assay of mouse bone marrow,sperm abnormality test and 30 days-feeding test were performed.Results All oral LD50 of the drug Wangla in male and female rats and mice were >10.00 g/kg body weight.It is actually non-toxic.All results of micronucleus assay of bone marrow,sperm abnormality test and genetic toxicity test were negative.No apaprent toxicity was shown by the 30 days-feeding test.Conclusion The Tibetan drug Wangla can be orally comsumped.

Abstract:目的建立一种快速检测H5N1病毒感染的血清学方法。方法以H5N1病毒作为抗原包被96孔板,与待检血清作用后,以辣根过氧化物酶标记山羊抗小鼠IgG作为二抗,用ELISA方阵法确定最佳抗原、血清稀释浓度。结果感染7 d小鼠血清中抗体水平很低,感染14 d血清抗体水平较高,最佳抗原稀释度1∶100,最佳血清稀释度1∶200。结论这种快速检测H5N1病毒的诊断方法直接有效、方便易行,有助于H5N1病毒感染的早期诊断

Abstract:目的应用Image-Pro Plus 5.0图像处理和分析软件,研究鸡胚尿囊膜(chick chorioallantoic membrane,CAM)血管新生面积定量的新方法。方法20只发育良好的7日龄鸡胚,分为龙葵给药组和对照组,每组10只。将中药龙葵水提液及等量蒸馏水吸附于5 mm直径的定性滤纸,置于CAM上。利用Image-Pro Plus 5.0软件,定量血管新生面积、蛋壳开窗处对应的CAM面积,计算出血管新生面积与CAM面积的比值。结果用Image-Pro Plus 5.0可方便、自动、准确地进行给药前后的数据收集和面积计算。统计分析表明,受试物龙葵可明显抑制CAM血管新生,与对照组比较有极显著差异(P<0.001)。结论Image-Pro Plus 5.0图像处理和分析软件是一种高效、准确的统计血管新生面积的工具,用该软件定量蛋壳开窗部位下的血管新生面积占开窗部位所对应的CAM总面积,不仅操作简便,而且数据计算自动生成,可较准确地反映鸡胚血管新生情况。

Abstract:Peripheral neuropathic pain is often difficult to be adequately treated in current medicine.Animal models may provide pivotal systems for preclinical study of pain.Partial spinal ligation,chronic constriction injury,and L5/L6 spinal nerve ligation represent three of the best characteristic rodent models of peripheral neuropathy.Recently,primary or permanent cultures of sensory neurons have been established to study the molecular mechanism of pain,especially for neurotransmitter release and signal transduction.

Abstract:Objective To investigate the influence of certain peculiar formula feed leading to a slow increase of body weight in laboratory animals.Methods To modify the formula of the peculiar feed reported in the literature,and to match the composition of various nutrients rationally,especially to raise the content of proteins.Results It was found that the body weight of the rats increased and returned to normal after feeding with the modified peculiar formula feed.Conclusion When using peculiar formula feed,the low protein contents may frequently be an important cause of a slow increase of body weight in laboratory animals.