Abstract:目的经尾静脉一次性注射博莱霉素(BLM)复制小鼠肺间质纤维化动物模型,并观察模型的肺组织病理学变化。方法8周龄雌性C57BL/6小鼠66只,随机分为BLM80组18只、BLM150组19只、BLM300组19只和对照组10只,分别经尾静脉一次性注射BLM80、150、300 mg/kg和生理盐水。结果①BLM80组和BLM150组小鼠最低体重分别为BLM注射前的84%和65%。②BLM80组、BLM150组、BLM300组和对照组小鼠的生存率分别为:100%、43%、0和100%。③BLM150组BLM注射后14 d、28 d,右肺羟脯氨酸含量分别为738±46 nmol、886±83 nmol,与对照组(360±75 nmol)比较差异均有显著性意义(P<0.01)。④BLM150组小鼠BLM注射28 d,在胸膜下及血管周围形成广泛、稳定、明显的纤维化改变。BLM150组与BLM80和对照组比较,肺纤维化病理评分分别呈明显增高(P值均小于0.001)。结论C57BL/6小鼠经尾静脉一次性注射BLM150mg/kg可以制备肺间质纤维化动物模型。

Abstract:目的采用三种方法建立兔单肺通气模型并比较其效果。方法日本大耳白兔30只,随机分为3组(即A、B、C组)各10个,分别采用自制双腔气管导管法、左主支气管结扎法和插管过深法。呼吸机通气参数为:FiO21.0,RR 40/min,VT 10 mL/kg。单肺通气2 h后恢复双肺通气。记录各组单肺通气实施的一次成功率、总成功率、从气管切开开始到单肺通气实施所需要的时间、动物失血量。实验结束后开胸测量兔气管、左主支气管、右主支气管的长度和内径。结果各组进入实验的动物数分别为10、6、8只。与B、C组比较,A组一次成功率和总成功率高,所需时间明显较少(P<0.01),且出血量明显少于B组(P<0.01)。结论采用自制双腔气管导管能迅速有效的建立单肺通气模型,是用于研究与单肺通气相关病理生理机制的理想模型。

Abstract:Objective The aim of this study was to explore the possibility to establish single-cell colonal lines from chicken embryonic primordial germ cells(EPGCs) and investigate their biological characteristics.Method single-cells were derived from the 4th passages and multiplied clonal EPGCs at stage 19 and 28,which were digested with 0.25% trypsin-EDTA.Then the single cell was picked up and seeded into individual well of a 96-well plate containing fibroblast feeder-layer,and every well contained one cell.The outgrowth of single-cell clones were passed after treatment with trypsin-EDTA,and their biological characteristics were detected by cytochemistry and histoimmunochemistry to detect their special markers.The karyotypes were assessed by routine test.The pluripotency was analyzed by differentiation in vitro.Results Nine in total of 288 clones maintained the normal diploid karyotype and undifferentiated.7 clones were passaged for 2 passages,and 2 clones were passaged for 4 passages.The 9 clones possessed normal karyotypes,expressing a series of surface markers such as: alkaline phosphatase and stage-specific embryonic antigen(SSEA)-1.Conclusion Nine single cell-cloned of chicken embryonic PGCs have been derived in our laboratory.The cells possess stable biological characteristics of undifferentiated PGCs.

Abstract:目的探讨兔肠道组织抗菌有效成分的组成及其性质。方法将新鲜兔小肠组织匀浆,经高温处理,乙酸浸提后,检测抗菌活性,再经Sephadex G100和Sephadex G75凝胶柱过滤层析,收集具有抗菌活性的蛋白组分,经SDS-PAGE电泳,考马斯亮蓝染色后,为一条蛋白带,其相对分子质量约为43×103。用琼脂糖弥散法和活菌计数实验检测纯化物对9株细菌的抑菌作用。结果分离得到一种纯的兔肠源抗菌蛋白,纯化的兔肠源抗菌蛋白对9株细菌的均有明显的抑菌作用,杀菌率介于78%与98%之间,显示了较强的抗菌活性。结论初步显示了兔肠源抗菌蛋白在细菌性疫病的防治方面的应用前景。

Abstract:Objective To study the effect of cartilage polysaccharide on S_(180)-bearing mice and its anti-tumor mechanism.Methods The S_(180)-bearing mouse model was prepared and treated by i.p.injection of cartilage polysaccharide.The tumor cells morphology was examined by light microscopy.The cell cycle was analyzed by flow cytometry,and apoptotic rate was determined by TUNEL assay.The expression of Fas and PCNA proteins was studied by immunofluorescent staining.Results The survival rate of S_(180)-bearing mice was significantly increased by cartilage polysaccharide treatment in comparison with that of control group.Cell shrinkage,nuclear condensation and apoptotic bodies were detected by morphological observation.The cell cycle of S_(180) cells was arrested at G_2/M phase. The expression of Fas protein was increased and PCNA expression was decreased at 24 h after cartilage polysaccharide treatment.Conclusion Cartilage polysaccharide induces apoptosis of S_(180) cells by cell cycle arrest and influences the expression of Fas and PCNA proteins,and inhibits tumor cell growth,and prolong survival time of S_(180)-bearing mice.Those findings indicate that cartilage polysaccharide might become a novel anti-tumor agent in future.

Abstract:目的应用B超技术对恒河猴子宫内膜周期性变化进行实时监测。方法采用Aloka 3.5 mHz探头的B超仪,分别对6只性成熟的正常雌性恒河猴进行连续3个月经周期(n=18)的子宫内膜的周期性变化的实时测量,同时进行连续2个周期(n=12)的血清雌二醇E2及孕酮P的放射免疫检测分析。结果恒河猴子宫内膜厚度变化呈周期性。在滤泡期(月经周期的第-9~0天),B超图像表现为子宫内膜区域相对子宫肌层区域回声较低(较黑),子宫内膜厚度呈线性增长(r=0.88);黄体期(月经周期的第0~17天),B超图像表现为子宫内膜区域光点增强为强回声,子宫内膜厚度无明显增长。子宫内膜周期性变化与血清E2及P相关,子宫内膜线性增长最大值出现时间与E2分泌最高峰出现时间相一致。结论B型超声监测技术(ultrasound examination,US)作为一种有用的手段,能实时监测恒河猴子宫内膜周期性变化。

Abstract:目的建立红色荧光和绿色荧光转基因小鼠,为活体荧光影像系统建立重要的实验动物模型。方法把DsRed-Express和EGFP基因插入chicken-βactin强启动子下游构建转基因载体,建立红色荧光和绿色荧光转基因C57BL/6J小鼠。PCR鉴定红色荧光和绿色荧光转基因小鼠的基因表型,活体荧光影像系统分析红色荧光和绿色荧光转基因小鼠,荧光显微镜检测红色荧光和绿色荧光转基因小鼠全身组织器官的组织形态。结果分别建立了3个系的红色荧光和3个系的绿色荧光转基因小鼠。活体荧光影像系统分析转基因小鼠分别呈现红色荧光和绿色荧光。经荧光显微镜观察,DsRed-Express转基因小鼠的红色荧光蛋白在多个组织器官中表达,尤其在胰腺、肝脏、肾脏和脾脏等器官表达量较高。EGFP转基因小鼠绿色荧光蛋白在全身各个组织器官中表达,尤其在胰腺、心脏、小肠、外周血细胞和脑组织等器官组织中表达量较高。结论DsRed-Express和EGFP基因在转基因小鼠中系统性高表达,成功建立了红色荧光和绿色荧光转基因小鼠。DsRed-Express和EGFP转基因小鼠将成为活体荧光影像系统的重要实验动物模型。

Abstract:Objective To study the sensitivity of Chinese non-human primates to H5N1 avian influenza virus and to establish animal models for H5N1 research.Method\ Cynomolgus and rhesus macaques were infected with H5N1 virus through nasal dropping.All monkeys were euthanized at various time points for collecting serum samples for serology,throat swab for etiology and tissues for pathology.Results\ After infection the animals manifested lack of appetite,transient fever and leukopenia.Virus replication was demonstrated by virus isolation and RTPCR.Histopathological examination of lung tissues at acute stage confirmed interstitial pneumonia,localized haemorrhage and necrosis,presence of inflammatory infiltration in the interstitium and interalveolar space.On the 14th day after infection,serum IgG level of the macaques went up.Conclusion\ Chinese rhesus and cynomolgus macaques can be infected with H5N1 virus and may serve as animal models for H5N1 research,providing better insight to the mechanism of AIV infection,and may greatly facilitate the screening of anti-AIV drugs and vaccine evaluation as well.

Abstract:目的观察15-酮基二十碳四烯酸(15-ketoeicosatetraenoic acid,15-KETE)对大鼠肺动脉平滑肌细胞(pulmonary arterial smooth muscle cells,PASMCs)膜电压门控钾离子通道(Kv)的作用,探讨其收缩肺动脉的离子通道机制。方法采用急性酶分离法(胶原酶Ⅰ型和弹性酶)获得健康成年SD大鼠单个PASMC,应用全细胞膜片钳记录方法,研究15-KETE对膜电位(Em)、膜电容(Cm)、电压门控钾电流(Ikv)的影响。结果①高浓度15-KETE(1×10-7mol/L、1×10-6mol/L)可引起PASMCs去极化,并且在细胞内钙被BAPTA缓冲后,15-KETE仍可引起PASMCs去极化,15-KETE对PASMC的膜电容无影响;②15-KETE(1×10-8~1×10-6mol/L)对Ikv的影响呈浓度依赖性和可逆性;③细胞内钙离子在生理浓度时([Ca2 ]i=75 nmol/L),15-KETE(1×10-6mol/L)对Ikv峰电流的抑制率显著高于细胞内无钙离子时。结论15-KETE可浓度依赖性的抑制Ikv,使常氧大鼠PASMCs去极化;细胞内钙离子加强了15-KETE对Ikv峰电流的抑制作用。

Abstract:目的研究17β-雌二醇(E2)暴露对雄性剑尾鱼(Xiphophorus helleri)卵黄蛋白原(vitellogenin,Vtg)诱导作用作为环境风险评价(ERA)的有效生物学标记的可行性。方法以E2诱导的剑尾鱼雄性个体的整体匀浆液为材料,采用Sephacryl S-300凝胶过滤层析柱和QSepharose阴离子交换柱从剑尾鱼体内提纯Vtg。结果确定了被纯化的剑尾鱼Vtg在4%~7.5%Native PAGE电泳中相对分子质量为540×103。4%~7.5%Native PAGE电泳后的凝胶分别利用苏丹黑B进行脂蛋白染色、高碘酸-Schiff试剂进行糖蛋白染色和甲基绿进行磷蛋白染色,表明剑尾鱼Vtg是一种富含糖、脂、磷的蛋白。结论表明雄性剑尾鱼卵黄蛋白原的诱导变化可作为环境风险评价(ERA)的有效生物学标记。

Abstract:Objective To observe the deposition of SIV specific immune complex in different organs at different time-points after SIV infection,and explore its possible relationship with multisystemic pathological changes in simian acquired immunodeficiency syndrome(SAIDS).Methods\ 8 SIV-infected rhesus monkeys and 1 uninfected monkey were autopsied and sampled.Serial sections were prepared,immune fluorescence staining of IgG,C3 and SIV p27 were performed,and the results were compared and analyzed.Results\ IgG,C3 and SIV p27 fluorescence expressed similarly at the same site in most monkeys,indicating the presence of SIV-specific immune complex.Among those positions,cerebral perivascular space(8/8),myocardial interstitial vessel(6/8),paracortex(5/8) and germinal center(5/8) of lymph nodes were the main sites with a higher positive rate.Their positive rates were elevated during the late phase after infection.Conclusion After SIV infection,viral specific immune complex is deposited widely,and increased remarkably with disease progression.SIV may lead to AIDS associated multisystem complication through IC deposition.Further research on it may be helpful to understand how SIV infection induces multisystem pathological changes,and it is also a promising target for developing new treatment strategy.

Abstract:目的研究脂多糖(Lipopolysaccharide,LPS)对实验性变应性鼻炎的影响。方法SD大鼠40只随机分4组,其中,变应性鼻炎组经腹腔注射及鼻腔滴入卵清白蛋白(OVA)致敏,建立变应性鼻炎动物模型;LPS刺激组经鼻腔滴入LPS(10μg/100μL);变应性鼻炎 LPS刺激组为大鼠激发成变应性鼻炎后再以LPS滴入鼻腔。观察各组的症状变化,如喷嚏,流涕等。行常规HE及甲苯胺蓝染色观察各组鼻黏膜炎性细胞的浸润情况,并行高倍镜下嗜酸性粒细胞计数。结果①变应性鼻炎 LPS刺激组过敏症状评分高于其余各组(P<0.01);正常对照组及LPS刺激组症状评分差异无显著性(P>0.05)。②变应性鼻炎 LPS刺激组鼻黏膜中嗜酸性粒细胞计数高于变应性鼻炎组,差异有显著性(P<0.05);正常对照组及LPS刺激组鼻黏膜中嗜酸性粒细胞计数差异无显著性(P>0.05)。结论LPS刺激可以加重变应性鼻炎的症状及鼻黏膜组织的病理学改变。

Abstract:目的急性肺损伤/急性呼吸窘迫综合征(Acute lung injury/Acute respiratory distress syndrome,ALI/ARDS)发病率与死亡率均较高,发病机制迄今尚不完全清楚,也无特效的治疗方法。本实验成功地建立一种轻型ALI动物模型,为研究该病的早期发病机制及治疗提供重要的观察手段。方法给予45只SD大鼠气管内灌注内毒素0.5 mL/kg(LPS 200μg/mL),观察4、12、24及48 h光镜和电镜下的病理改变;观察支气管肺泡灌洗液(BALF)中细胞分数、白蛋白等。结果在观察时间内实验动物均存活。LPS给予后实验组病理检查发现①肺间质水肿;②肺泡腔内多形核中性粒细胞(PMN)浸润和红细胞渗出;③肺泡Ⅰ型和Ⅱ型肺泡上皮细胞破坏。以LPS给予后4~12 h为最严重。BALF中PMN及白蛋白明显增加。结论气管内灌注内毒素0.5 mL/kg(LPS200μg/mL)成功地建立急性肺损伤动物模型。

Abstract:目的建立一种改良的新生兔缺氧缺血性脑损伤(HIBD)动物模型。方法选择孕期30 d的孕兔,随机分为正常对照组、缺氧5 min组、缺氧10 min组和缺氧15 min组。给予孕兔吸入7%二氧化碳的氮气使其窒息后剖宫产出新生兔,观察新生兔出生时的一般情况,4 d后作头颅磁共振影像(MRI)检查,5 d后处死动物采用HE染色和光镜观察新生兔脑组织结构改变,并作病理评分。结果缺氧10 min组新生兔活体观察、头颅MRI、病理改变符合窒息后HIBD动物的变化特点,MRI检查新生兔脑组织可见大片状、弥漫性分布的不均匀信号,呈稍短T2信号,白质、灰质界限模糊;正常对照组、缺氧5 min组和缺氧10 min组病理评分分别为(4±0,5.44±1.13,13.3±2.39),缺氧5 min组与正常对照组差异无统计学意义(P>0.05),缺氧10 min组主要见变性、坏死和小胶质细胞增生改变,与正常对照组差异有统计学意义(P<0.001),缺氧15 min组新生兔生后6 h内全部死亡,不作MRI检查及病理评分。结论向孕兔输7%二氧化碳的氮气10 min使其窒息后剖宫迅速取出新生兔是简单、快速、可靠制备HIBD模型的方法。

Abstract:Objective To develop a multiplex polymerase chain reaction(PCR) method for identification of RR-B strain of Xiphophorus helleri.Methods 6 pairs of special microsatelites primers,consisted of Msa012,Msa014,Msa033,Msb025,Msd003 and Msd051,were selected to perform multiplex PCR to distinguish RR-B swordtail fish strain from wild swordtail fish.Result Stable amplified product with primer series 1(Msa014,Msb025,Msd003) and series 2(Msa012,Msa033,Msd051) could be obtained.Those amplified segments of multiplex PCR could be used to clearly distinguish the strains from each other.The exclusion probability of primer series 1 was 99.98% and the number of primer series 2 was 99.96%.Conclusion The microsatelites multiplex PCR mothed developed in this study can be used to accurately and rapidly identify the RR-B strain of Xiphophorus helleri.

Abstract:目的研究山羊卵母细胞减数分裂过程中线粒体的动态分布。方法收集山羊卵母细胞,在M199中分别培养4、8、12、16、20和24 h,用特异性线粒体标记探针进行标记,用激光扫描共聚焦显微镜观察线粒体的分布情况。结果生发泡期线粒体多分散在卵母细胞的胞质内,并且距生发泡有一定的距离;生发泡破裂期线粒体逐渐移向染色质;第一次减数分裂中期与第二次减数分裂中期线粒体成簇密布在染色体周围。排出的第一极体中也含有大量的线粒体。结论同其他哺乳动物卵母细胞体外成熟过程中线粒体分布情况相比,线粒体在山羊卵母细胞中的分布具有明显的相似性。线粒体密布在成熟卵母细胞染色体周围可能与极体的排出和受精后染色体的迁移有关。

Abstract:Objective To study post-testis targets related to male contraception and infertility caused by dutasteride treatment on rat sperm maturation and fertility.Methods Rats were treated with 20 and 40 mg/kg/d dutasteride for consecutive 14 days.Each male rat was paired with 2 female rats and the reproductive indices were assessed.Sperm motility and morphology were analyzed by a computer-assisted sperm analysis system.The sperm survival rate was assessed by SYBR-14 and PI fluorescent staining.Serum dehydrotestosterone(DHT) and T of rats were detected by ELISA.Histological examination of testis and epdidymis was conducted by HE staining.Results The DHT level,sperm motility and the pregnancy index by treatment were all significantly decreased(P<0.05 or 0.01),while the abnormal sperm rate was increased(P<0.05).But there was no difference in serum T level between dutasteride treated and control males(P>0.05).The histology of testis and epididymis was also unaffected by treatment.Conclusions Dutasteride can induce infertility in male rats through inhibiting serum DHT level and interfering with epididymal sperm maturation,which may open a new approach for research on male contraception and fertility.

Abstract:Objective To detect the information of gastric emptying and infection of Helicobacter pylori for 30-weeks diabetic rats.Methods To establish 30 weeks diabetic rat model with Streptozotocin(STZ),then phenolsulfophthalein test was used for gastric emptying and infection of Helicobacter pylori was detected.Results The test showed that anormal gastric emptying existed in the 30-weeks diabetic rats.The Chinese medicine "Tangweikang" group improved compared with diabetic model group,Helicobacter pylori was negative.Conclusions In 30-weeks diabetic rats exists anormal gastric emptying without infection of Helicobacter pylori.

Abstract:Objective Canine transmissible venereal sarcoma(CTVS) is a naturally occurring sexually transmitted neoplasm of the genital system in dogs.This tumor is unique in oncology because it can be applied in homotransplantation and xenotransplantation.This review was designed to describe the biological features of CTVS cells and investigate the approach of CTVS tumor model.CTVS tumor model is reliable and can be developed in various tissues which resemble well the characteristics of the tumor.It can be applied in large animals and some organs difficult to be transplanted,provides a basis for a wide range of clinical and radiological oncology researches.