Abstract:Objective To investigate the mechanism(s) of dopamine D_5 receptor regulation to hypertension involved in the antioxidant actions.Methods We investigated blood pressures in D_5 receptor deficient(D_5-/-) and wild type(D_5 / ) mice,and NADPH oxidase protein expression(p47~(phox)) and activity in kidney of those mice.We also checked super oxide and hydrogen peroxide production by using HEK-293 cells heterologously expressing human D_5 receptor.Result The systolic,diastolic and mean arterial blood pressures,and NADPH oxidase activity and p47~(phox) expression were higher in D_5-/-than D_5 / mice.In HEK-293 cells heterologously expressing human D_5 receptor,its agonist,fenoldopam,decreased super oxide and hydrogen peroxide production.Conclusion\ The ability of D_5 receptor stimulation to inhibit NADPH oxidase activity so as to decrease ROS production may explain,in part,the antihypertensive action of D_5 receptor activation.

Abstract:Objective To analyze the polymorphisms of 20 microsatellites in 11 strains of mice by PCR and to explore a new method for inbred strain mice genetic monitoring.Methods 20 short tandem repeat(STR) primers were used to amplify DNA in 11 inbred strains of mice(C3H,DBA,BALB/c,C57BL/6,C57BL/6-Bg,C57BL/10,TA1,TA2,T739,M615 and SCID).The polymorphisms information of C57BL/6-Bg,C57BL/10,TA1,TA2,T739,M615 and SCID were acquired according to those of C3H,DBA,BALB/c and C57BL/6.Results 193 out of the 220 amplified bands were in anticipation.Conclusion The results showed that different stains of mice could be differentiated by use of polymorphisms of the 20 microsatellites.It revealed that genetic diversity is abundant in inbred mice.These results have instructive significance for mice genetic monitoring.

Abstract:目的建立巴马小型香猪的药物代谢研究模型,了解巴马小型香猪机体对药物的作用。方法选择抗动脉粥样硬化的HMG-CoA还原酶抑制剂型药物洛伐他汀(LV)为模型药,给6月龄雄性巴马小型香猪二组共8头,单剂量(6.0 mg/kg,2.4 mg/kg)灌胃,采用RP-HPLC法检测LV的血药浓度,并计算药代动力学参数。结果LV在巴马小型香猪的处置符合一室开放模型;Tmax分别为(2.54±0.18),(2.82±0.44)h;T1/2α分别为(1.53±0.05),(1.52±0.44)h;T1/2β分别为(2.04±0.32),(2.77±0.89);Cmax分别为(28.86±3.76),(10.93±1.77)ng/mL;AUC分别为(202.76±41.98)和(104.86±16.44)(ng/mL)×h。结论LV在巴马小型香猪的吸收速度约为正常人的0.5倍,消除速度为正常人的1.5~2倍,但整个代谢过程类似正常人,均属一室开放模型,提示巴马小型香猪可作为心血管系统药物LV药代研究的良好模型。

Abstract:Objective To study the effect of FSH and LH on superovulation in KM mice.Method 48 mice were divided into 4 groups,which received FSH 3 IU,6 IU,9 IU and 12 IU i.p.,respectively,quadrifidly and in a decreasing sequence.48 hours later,LH in a dose of 3 IU,6 IU,9 IU and 12 IU was injected.The female mice were combined with male mice in one cages,and were checked for embolus the next morning.Embryos were washed out 48 hours later.Result The rate of embolus visualization was 79.17%(38/48).The average of washing out embryos was 5.67(68/12),13.00(156/12),29.17(350/12),11.50(138/12) embryos per mouse according to the used hormones: 3 IU FSH 3 IU LH,6 IU FSH 6 IU LH,9 IU FSH 9 IU LH and 12 IU FSH 12 IU LH,respectively.Conclusion The protocol using 9 IU FSH 9 IU LH is the most effective to induce superovulation in KM mice among all schemes used in this study.

Abstract:Objective To establish an animal model of gastrointestinal microflora imbalance induced by an antibiotics,ceftriaxone.Methods Specific-pathogen-free mice received orally antibiotics in the drinking water ad lib(the concentration was 0.5 mg/mL),or via intragastrical gavage in a dose of 125 mg/mL,0.2 mL b.i.d.for 4 days.On the 5th day,the mice were sacrificed and intestinal flora analysis was performed.Results Both the two methods induced alteration of intestinal flora,but the intestinal mucosa of mice treated by intragastrical gavage showed more rapid repair and the translocation of bacteria was lower than that in the mice of group administered with antibiotics in drinking water.Conclusion Administeration of ceftriaxone in drinking water ad lib is a perfect method to induced intestinal microflora dysbiosis in mice.

Abstract:目的研究镇痛实验中以痛阈值作为检测麻醉效应指标的可行性及复方亚甲蓝注射液的镇痛效应.方法 100只小鼠随机分为5组：生理盐水组、利多卡因组、复方亚甲蓝注射液高、中、低剂量组.将小鼠固定于掌心,两电极分别刺于小鼠尾尖1/3处及左下肢足部,开通测量仪（量程0.5～1 mA,阶程6）,观察记录发生第一嘶叫反应的电流值,作为动物给药前之基础痛阈值.选用4^#针头随机区组设计注射于小鼠足跟偏内上部胫骨周围间隙内,再按照基础痛阈测定同样抓取小鼠并同部位刺激,观察记录给药后10 min、30 min、1 h、2 h、4 h、6 h、8 h、10 h、12 h、24 h、26 h之第一嘶叫反应的电流值,以电流的变化值及实际电流值为观察指标.结果利多卡因给药后10min痛阈值即显著升至最高点,其后逐步降低,其测定痛阈值与基础痛阈值及生理盐水组的同点测定痛阈值比较有极显著性差异的维持时间2～3 h左右,4 h后基本无镇痛效应;利多卡因给药后10 min、30 min、1 h、2 h痛阈相对基础痛阈的变化值与生理盐水组的同点测定痛阈相对基础痛阈的变化值比较有极显著性差异,维持时间2～3 h左右,4 h后变化值无统计学的意义;药后2 h～26 h各复方亚甲蓝组动物的痛阈值与利多卡因组比较有极显著性差异（P〈0.01或P〈0.05）.结论 2%利多卡因（0.05mL/只）镇痛作用持续时间小于4 h,而0.33%、1.1%和3.3%的复方亚甲蓝镇痛作用持续10～26 h.结果证明复方亚甲蓝具有及时而持久的局部镇痛效应.

Abstract:目的建立小鼠慢性心肌炎实验动物模型,为心肌炎发病机理的研究以及治疗药物的筛选提供一个可靠的实验手段.方法采用BALB/c小鼠500只,用驯化的柯萨奇B3病毒进行感染,在实验的第10天、20天、40天、80天和120天对实验小鼠进行各项实验数据检测.主要包括死亡率、病理形态、超微结构、心电图和血清特异性抗体等.结果①实验组小鼠死亡率为27.8%.②小鼠心脏重与体重比值随着实验时间的延长有明显的增加.③小鼠平均心肌炎发病率为49%,第120天仍有心肌病理改变.④实验小鼠心肌超微结构有明显改变.⑤实验小鼠异常心电图发生率约在50%以上.第120天实验小鼠心电图改变仍占57.7%.⑥小鼠血清特异性抗体IgG在各时间段变化不明显,而IgM随时间的延长阳性率有明显下降趋势.结论通过实验显示,一个稳定可靠的慢性小鼠心肌炎模型基本建立成功,本实验支持慢性心肌炎可导致扩张性心肌病的发生.

Abstract:目的对比测定贵州小型香猪与人肝微粒体细胞色素CYP酶的五个主要亚型的活性,并对二者活性相似的亚型以特异性抑制剂进行抑制,观察抑制活性反应的相似性,比较二者的CYP酶特性,评估贵州小型香猪作为人体外药物代谢动力学研究实验动物模型的可行性及其可应用的药物代谢反应类型.方法制备贵州小型香猪和人的肝微粒体,构建肝微粒体和探针底物的反应体系以及抑制剂抑制反应的体系,利用HPLC测定CYP3A,CYP1A,CYP2A,CYP2D和CYP2E酶的探针底物与肝微粒体反应的活性,并对贵州小型香猪和人的相应活性值进行比较,选择活性最为相近的亚型进行特异性抑制.结果贵州小型香猪肝微粒体具有CYP酶的五个主要亚型的反应活性,其中CYP3A酶的两个特异反应即硝苯地平氧化反应,睾酮6β-羟化反应的活性值和人肝的相应活性接近.同时,二者的CYP3A酶的特异性抑制反应相似.结论贵州小型香猪适用于作为人的CYP3A酶及其相关药物代谢研究的良好动物模型.鉴于CYP3A酶是人体内Ⅰ相反应最主要的药物代谢酶,因此贵州小型香猪作为人体药代动物模型具有一定前景.

Abstract:Transmission of viral infection by tumour lines or other biological materials may have confounding effects on research Many research organizations require screening for viral agents of all cell lines, tumours, sera and other biologicals before implantation or inoculation into animal models. Screening for viral contamination is done by the mouse antibody production （MAP） test, by cell culture, or alternatively by direct detection of the viral agents by polymerase chain reaction （PCR）. The description of procedures for sanitation of infected cell lines or tumours is sparse. The present report describes the procedures used for sanitation of three transplantable murine tumour lines, which were transplanted in vivo in a mouse hepatitis virus （MHV）-infected colony of mice at the Department of Experimental Clinical Oncology （DECO）. The tumours were frozen and serially transplanted three times in a quarantine colony of syngenic mice.

Abstract:目的大鼠在屏障环境和独立通气笼盒（Individually ventilated cages,IVC）中饲养条件下,某些生物学特性变化的比较.方法分别将10对SD大鼠在屏障环境和独立通气笼盒中饲养繁殖,观察生长发育和繁殖性能,测定50日龄仔鼠血常规、血生化指标.结果 （1）大鼠生长发育及繁殖性能中胎间隔差异无显著性（P＞0.05）;屏障环境中雄性大鼠在30 d前较IVC中雄鼠增重明显,40d后差异不显著;雌鼠在30 d以后差异显著.（2）血常规指标中血小板计数（PLT）、血小板比积（PCT）、血小板分布宽度（POW）、平均血小板体积（MPV）、大血小板（LPCR）差异有显著性（P〈0.01）;血生化中总胆红素（TBil）、直胆红素（DBil）、间胆红素（IBil）、碱性磷酸酶（ALP）、肌酐（Cre）、葡萄糖（Glu）、铜（Cu）、锌（Zn）、钠（Na）、氯（Cl）、总二氧化碳（CO2）升高（P〈0.05或P〈0.01）,乳酸脱氢酶（LDH）、磷酸肌酸激酶（CK）、磷酸肌酸激酶同工酶（CK-MB）、低密度脂蛋白胆固醇（LCL-C）、载脂蛋白B（ApoB）、脂肪酶（LPS）、镁（Mg）下降（P〈0.05或P〈0.01）.结论在屏障环境和IVC条件下,大鼠的生长发育及某些血液学指标有一定的差异.

Abstract:Bladder cancer is regarded as a promising candidate for innovative therapies in the field of immune and gene therapy. In this paper, we present the subcutaneous, metastatic and a novel orthotopic model of murine MB49 bladder cancer in C57BL/6 mice. We further show the potential of using adenoviral vectors together with different transduction enhancers to augment in vivo gene delivery. Finally, we present candidate genes for tumour detection, therapy or targeting. The MB49 tumour grew rapidly in mice. The subcutaneous model allowed for tumour detection within a week and the possibility to monitor growth rate on a day-by-day basis. Injection of MB49 cells intravenously into the tail vein gave rise to lung metastases within 16 days, while instillation of tumour ceils into pretreated bladders led to a survival time of 20 ～ 40 days.

Abstract:Objective To assess the influence of cysticercus cellulosae cell vaccine on immunization in pregnant sows,piglets with maternal antibody and normal piglets.Mothods\ To measure serum antibody by ELISA method and determine the immunized results of different experimental pigs.Results\ Antidody titer of pregant sows reached a titer of 1:20480.Piglets possessed maternal antibody showed a near the effective titer 1:2560.Giving an additional immunization after birth at proper time,the antibody level can be further raised shortly.Conclusion\ The maternal antibody from pregnant sows immunized by Cysticerus cellulosae cell vaccine can be transmitted to piglets,making them possessing congenital immunity and resistant to invasion of Cysticercus cellulosae eggs.Immunization of normal piglets with the same vaccine induces an effective antibody level much slowly.

Abstract:目的研究放射治疗诱发BALB/c小鼠Renca肾癌细胞凋亡的特点及其调控机制,为肿瘤放射治疗提供实验依据。方法采用Annexin-V-FITC/PI法流式细胞仪检测不同剂量放疗后细胞凋亡率,建立BALB/c小鼠肾癌细胞肿瘤种植模型,采用PV免疫组织化学法检测肿瘤石蜡切片P53的表达及ELISA试剂盒检测细胞因子IL-2、IFN-γ的表达。结果①在一定照射剂量范围内,Renca细胞的凋亡率随剂量的增加而升高。②放疗组突变型P53蛋白表达率为(30.17±0.88)%,肿瘤组为(24.22±0.68)%,两组间有显著的统计学差异。③放疗组小鼠脾细胞分泌IL-2为(167.56±13.13)pg/mL,为肿瘤组小鼠分泌量(83.69±1.37)pg/mL的2倍,两组间有显著的统计学差异。放疗组小鼠脾细胞分泌IFN-γ为(1482.47±47.91)pg/mL,约为肿瘤组小鼠分泌量(211.02±35.98)pg/mL的7倍,两组间有显著的统计学差异。结论放疗可诱发Renca细胞凋亡,P53蛋白可能涉及本实验中的细胞凋亡通路。

Abstract:Objective VP1 protein of foot-and-mouth disease virus(FMDV) is the most antigenic protein being responsible for eliciting neutralizing antibodies and conferring protective immunity.The successful expression of VP1 gene in Bac-to-Bac expression system should lay a foundation for research of FMDV Type O serology method.Methods\ We inserted VP1 gene of Foot-and-Mouth virus type O into pFastBacHIa vecter to get recombinant plasmid pFastBacHIa-VP1.The recombinant plasmid was transferred into DH10Bac containing a shuttle vector Bacmid and the VP1 gene was integrated into Bacmid by site-specific transposition.Subsequently, the recombinant shuttle vector Bacmid-VP1 was transfected with Sf9 cells.Results\ SDS-PAGE electrophoresis and Western blot analysis detected a band of about 26.4 kDa protein in the expression product of VP1 gene in insect cells.Conclusion\ The successful expression of VP1 gene in Bac-to-Bac expression system should lay a foundation for research of antigenicity and immunogenicity of VP1 protein as well as the surveillance of antibody.

Abstract:目的以小鼠卵母细胞为模型,利用开放式拉长细管(open pulled straw,OPS)法研究在不同条件下冷冻、解冻对其发育能力的影响,为家畜、濒危动物乃至人类探索一种简易、高效的卵母细胞冷冻保存提供理论与技术参考。方法在室温(25℃±0.5℃)条件下,以乙二醇(EG)、DMSO为主体抗冻保护剂配制成的玻璃化溶液(EFS、EDFS),采用OPS法对卵母细胞进行玻璃化冷冻保存。实验一:操作台温度分别为25℃±0.5℃和37℃±0.5℃时,卵母细胞在10%EG 10%DMSO或10%EG溶液中预处理30 s,移入EDFS或EFS中平衡15 s~45 s进行冷冻保存;实验二:操作台为37℃±0.5℃条件下,采用三种方法(卵母细胞在A:0.5 mol/L和0.3 mol/L的蔗糖中分别处理2 min和5 min;B:0.3 mol/L和0.15 mol/L的蔗糖中分别处理5 min和2 min;C:0.5 mol/L的蔗糖处理5 min)对EDFS30和EDFS40冷冻保存的卵母细胞进行解冻;实验三:解冻后的卵母细胞在SrCl2溶液中进行孤雌激活。结果与结论实验一中卵母细胞解冻后的形态正常率分别高达92.1%(25℃±0.5℃)和92.5%(37℃±0.5℃),均与对照组(98.9%)无显著性差异(P>0.05);实验二中卵母细胞解冻以C法效果最佳(形态正常率分别为92.5%和67.5%);实验三中解冻后在mCZB中培养0.5 h的卵母细胞激活效果优于未培养组(卵裂率和囊胚发育率分别为80.0%vs.57.7%;18.2%vs.0),均有显著或极显著差异(P<0.05或P<0.01);而培养组与新鲜对照组卵母细胞激活后卵裂率和桑椹胚发育率与对照组(80.0%vs.77.8%;52.8%vs.61.9%)均无显著性差异(P>0.05)。

Abstract:目的探讨剑尾鱼肝细胞原代培养的方法。方法分别用机械分散法、酶消化法对肝细胞进行分离,并比较在William’s E(Williams’Medium E)、DMEM、DMEM/F12无血清培养基中肝细胞的生长效果。结果⑴组织块培养法(机械分散法)的肝细胞在William’s E、DMEM、DMEM/F12无血清培养基中,温度25℃~28℃,pH 7.3左右时都可以短期正常生长,而生长效果好依次是William’s E、DMEM、DMEM/F12无血清培养基。⑵酶消化法分离的肝细胞的产量为(6.4±1.7)×106 cell/g肝组织,即时存活率为(85±6)%。24 h贴壁率不足20%。结论酶消化法培养肝细胞产量、即时存活率及贴壁率偏低。组织块培养法肝细胞生长良好,而不需要苛刻的条件,可以满足毒理学实验要求。培养肝细胞首选William’s E培养基。

Abstract:Development of high-throughput gene microarray as a new powerful technique has improved our understanding of the rheumatoid arthritis, and it has been successfully applied to study the inflamed paws of mice or rats systemically immunized with arthritogenic compounds to induce arthritis. This review highlights present and potential applications of gene microarray technology for animal models in research of rheumatoid arthritis.