Abstract:Objective To investigate whether synaptic NMDA receptor activation induces Wnt non-canonical signaling pathway activation.Methods Various methods such as primary cortical neuron culture,immunofluorescence staining,double immunofluorescence staining,and Western blotting combined with NMDA activation were used to assess the increase of some proteins in Wnt non-canonical pathway.Results The methods of primary cortical neuron culture were established and the purity of neurons was enough for experiment.Wnt5a was...

Abstract:目的 摸索出一套副溶血性弧菌基因敲除的可靠方案,副溶血性弧菌致病相关基因的敲除对深入研究其致病机制有重要意义.方法 通过融合PCR技术将目的 基因上下游同源臂融合并克隆到自杀载体pDS132上,将重组质粒转化大肠杆菌S17λpir中,再接合转移到副溶血性弧菌菌株内,经pDS132质粒上sacB基因的反向筛选得到突变株.结果 成功构建了副溶血性弧菌RIMD2210633菌株ΔopaR,ΔtoxR和ΔaphA三个基因突变株.结论 通过自杀载体同源重组成功获得精确敲除的无痕突变株更有利于基因功能的研究,使后续副溶血性弧菌突变株与野生株的对比研究成为可能.

Abstract:目的 确定DEAE-葡聚糖对CEMx174细胞的半数抑制浓度,明确其在SHIV病毒TCID50滴定及病毒扩增中的促进作用.方法 分别使用含DEAE和无DEAE的DMEM完全培养基测定SHIVchn19p7的TCID50.用无血清DMEM培养基系列稀释DEAE,加入CEMx174细胞,使用cck-8测定细胞破坏率.分别选取DEAE浓度为28.125μg/mL和14.0625μg/mL的无血清DMEM培养基对CEMx174细胞预处理3 h.再加入SHIV-KB9病毒液,定期测定培养上清中的P24水平,同时做正常病毒对照和DEAE-1640对照,比对不同处理下的病毒扩增情况.结果 使用了DEAE后,SHIVchn19p7的TCID50达到了3.16×104TCID50/mL,不使用DEAE,病毒的TCID50测定为阴性.DEAE对CEMx174细胞的IC50为44.85μg/mL.经浓度为28.125μg/mL和14.0625 μg/mL的DEAE预处理后,SHIV-KB9病毒扩增在13 d～17 d达到高峰.而用不含DEAE的1640生长液培养的实验孔在19 d才开始出现阳性反应.结论 高浓度的DEAE对细胞有较强的杀伤作用,低浓度的DEAE对细胞的破坏率较低,并且能显著促进病毒扩增.DEAE在病毒进入细胞的过程中确实起了重要的作用.

Abstract:Objective To evaluate the sellection of suitable internal reference genes for normalization of gene expressions in mouse.Methods The expression levels of six housekeeping genes including GAPDH,HPRT1,B2M,PPIA,ACTB and 18S rRNA in the hypothalamus,pituitary and ovarian tissues and two developmental stages of C57BL/6J and C3H/HeJ mouse strains were assessed by quantitative real-time PCR(qPCR).Results The variation of expression levels was analyzed using geNorm program and demonstrated that GAPDH and HPRT1 were...

Abstract:目的 为兔骨骼肌匀浆多点皮下注射制备多发性肌炎大鼠模型的方法探索最佳的实验对象和实验条件.方法 SD大鼠、Westar大鼠和Lewis大鼠根据品种、性别、免疫物剂量等因素分组建模,取股四头肌进行组织病理学检查,比较各组之间成模比例、死亡比例、肌肉病理评分等观察指标的差异.结果 ①三个大鼠品种中,Lewis大鼠模型成功率最高,且实验耐受性最好,相同免疫剂量组的死亡比例最低.②相同免疫剂量下雌性组的成模比例均高于雄性组,而肌肉病理评分在不同性别组之间差异不明显.③雌性Lewis大鼠中,注射兔骨骼肌匀浆蛋白剂量从8 mg/kg增加至10mg/kg时,成模比例及模型病理评分明显升高;但从10mg/kg增加至12 mg/kg时,成模比例及模型病理评分无明显变化,而大鼠死亡比例升高.④雌性Lewis成模大鼠中,免疫2周时即可见到骨骼肌纤维大小不一、横纹消失及核内移,免疫4～5周时可见骨骼肌纤维坏死、非坏死肌纤维及间质血管周围炎细胞浸润等典型病变,继续免疫至7周时骨骼肌病变未继续加重.结论 采用兔骨骼肌匀浆多点皮下注射方法制备多发性肌炎大鼠模型时,选择雌性Lewis大鼠作为实验对象,给予兔骨骼肌匀浆10mg/kg连续免疫5周可出现与人类多发性肌炎非常相似的骨骼肌病变,且成模比例高、死亡比例低,造模效果理想.

Abstract:Objective To select and establish suitable experimental animal models of skeletal fluorosis and analyze their sensitivity to fluoride-induced bone damages.Methods New Zealand rabbits,Hartley white guinea-pigs,Wistar rats,and Kunming mice(16 each) were included in this study.Each kind of the animals were divided randomly into two groups(each group 8): control group(group C) fed with standard food and distilled water,and model group(group M) fed with normal food and 150 mg/L NaF distilled water).At 60 days af...

Abstract:Objective To co-infect miniature pigs with influenza A virus A/California/7/2009 with A/Brisbane/10/07 and A/Shenzhen/406H/06 virus,respectively,and to verify whether the co-infection of A/California/7/2009(CA7) and A/Brisbane/10/07(H3N2) or A/California/7/2009 and A/Shenzhen/406H/06(H5N1) influenza viruses will induce mutation in influenza virus.Methods To co-infect the 5-6 month old miniature pigs with A/California/7/2009(CA7) and A/Brisbane/10/07(H3N2) or A/California/7/2009 and A/Shenzhen/406H/06(H5N1) ...

Abstract:Objective To investigate the changes of TLR4 and CD14 mRNA expression in the rats with acute lung injury(ALI) induced by lipopolysaccharide(LPS).Methods Twenty-four male Wistar rats were randomly divided into two groups: normal control group and LPS model group.Each group had two subgroups,4 hours and 8 hours after LPS injection.The ALI model was established by intravenous injection of LPS(10 mg/kg) through the tail vein.The arterial blood gas graph(ABGG) and lung-body index were analyzed.The expressions of...

Abstract:目的 研究羊骨胶原肽(sheep bone collagen peptide,SBCP)对类固醇诱导的去卵巢大鼠催乳素(prolactin,PRL)和促黄体生成素(luteinizing hormone,LH)分泌脉冲的影响.方法 6周龄子宫颈未开口的青春期SD大鼠实施双侧卵巢摘除手术,康复1周或3周后以类固醇替代方法诱导PRL和LH脉冲,处理组同时按1000 mg/(kg·d)的剂量以SBCP灌胃,通过颈导管采集血样,利用放射免疫技术测定各组大鼠外周血中的LH和PRL浓度.结果 对于术后康复1周的大鼠,SBCP对LH脉冲振幅起到增强作用,而对PRL脉冲没有影响;对于术后康复3周的大鼠,SBCP对LH和PRL脉冲振幅均表现为降低作用,但卵巢摘除后立即给予SBCP可减弱这种降低作用.结论 雌激素和孕激素替代注射的同时,补充羊骨胶原肽,不仅仍可诱导LH和PRL脉冲的产生,还可预防由于卵巢摘除带来的骨代谢紊乱,免疫力低下等不良影响,因而是对现有类固醇替代方法的改良.

Abstract:Objective To determine the genetic variation and genetic relationship between the wild mouse captured from Lankao county,Henan province,and four common strains of laboratory mice(B6,BALB/c,DDK,PWK),and to construct a wild-origin inbred line mouse.Methods Totally 25 pairs primers were used to identify DNA polymorphisms that distinguish the five strains of mice by amplified fragment length polymorphism(AFLP).Results AFLP primer combinations detected a total of 2035 amplification bands.There were 507 polymorph...

Abstract:Objective To observe the pathological changes in the livers of captive rhesus monkeys,and explore their distribution rule and provide basic background reference data for relevant biomedical studies.Methods One hundred and fifty five natularly dead rhesus monkeys were recorded in our department from 1998 to 2008 years.Autopsy was performed and the pathological changes in the livers were examined and statistically analyzed according to different ages at 2 to 20 years.Results Of the 155 monkeys,88 animals had ...

Abstract:Objective To compare the effects of different insertion depths of nylon thread on the establishment of rat model of middle cerebral artery occlusion(MCAO).Methods The rats were randomly divided into four groups according to different insertion depths of the nylon thread:(1) 0.8 cm group;(2) 1.3 cm group;(3) 1.8 cm group;(4) 2.2 cm group.The body weight and modified neurological severity scores were tested before MCAO,24 h and 48 h after MCAO.All rats were killed at 48 h after MCAO.The brains were removed an...

Abstract:Objective To establish a model of early atherosclerosis in guinea pig and explore its pathogenetic mechanisms,compared with the modeling in rats,and to clarify the characteristics and advantages of this guinea pig model of atherosclerosis.Methods The differences of early atherosclerosis induced by high lipid diets in guinea pig and rat were investigated.Aortic intima-media thickness,intimal inflammatory cells infiltration and plaque development were observed by pathology with HE staining.Serum lipid levels ...

Abstract:Objective To study the influence of weichangning,a Chinese medicine decoction,on rat models of experimantal spleen deficiency syndrome.Methods Thirt-six male Wistar rats were randomly divided into 4 groups: normal control group,spleen deficiency syndrome model group,Sijunzitang(another Chinese medicine decoction) group and Weichangning group,9 rats in each group.The rat models of spleen deficiency syndrome were induced by resperpine.The plasma concentrations of LDH,AKP,AchE and CK and the contents of SOD,MD...

Abstract:Objective To compare four methods of bacterial genomic DNA extraction from Sprague-Dawley rat fecal samples.Methods Sixteen fecal samples with 106 cfu S.aureus and 4 fecal samples without S.aureus were equally divided into four groups.Bacterial genomic DNA were extracted by following methods:(1) Several wash by PBS plus phenol/chloroform purification;(2) Liquid nitrogen digestion plus column purification;(3) PBS wash plus column purification,and(4) Liquid nitrogen digestion followed by phenol/chloroform pur...

Abstract:Objective To investigate the feasibility of heparin-bonded biodegradable polycaprolactone(PCL) materials in animal femoral artery transplantation.Methods Heparin-bonded biodegradable PCL materials loaded with basic fibroblast growth factor(b-FGF) were used as vascular tissue engineering scaffold,and small-diameter tubular scaffolds for tissue engineering blood vessel were constructed using electrospinning technique.Endothelial cell adhesion experiment was conducted to evaluate the adhesion of endothelial ce...

Abstract:目的 评价聚乳酸聚乙醇酸膜、聚β羟基丁酯膜和胶原膜的结构、生物相容性及其在组织工程血管中的应用前景.方法 HE、胶原染色,扫描电镜观察材料的结构.新西兰兔15只皮下植入材料,于4、6、8和12周取出观察其组织反应和降解情况.将犬股动脉间质细胞种植于聚乳酸聚乙醇酸膜、胶原膜上,观察其形态.结果 聚乳酸聚乙醇酸膜、聚β羟基丁酯和胶原膜的孔径分别为179、13、107 μm,3种材料组织相容性好,局部无组织坏死和脓肿形成.早期以中性粒细胞和嗜酸性粒细胞浸润为主,后期以淋巴细胞浸润为主.聚乳酸聚乙醇酸膜于8周基本吸收,胶原膜12周吸收,聚β羟基丁酯膜12周未见明显降解吸收.细胞种植实验显示细胞在聚乳酸聚乙醇酸膜和胶原膜表面生长良好.结论 聚乳酸聚乙醇酸膜和胶原膜在材料结构、组织及细胞相容性和降解率方面符合组织工程血管支架材料的要求.聚β羟基丁酯膜在材料结构、降解率和细胞相容性方面尚需改进.

Abstract:Objective To observe the changes of the biological characteristics and hormonal alterations in hypophysectomized immature rats.Method Eighty 4-weeks old Sprague-Dawley young rats underwent hypophysectomy via parapharyngeal approach within two days,and among them 20 qualified operated rats were taken as the group A(experimental group).They were raised together with 20 control rats(group B) under the same conditions.Their body weight,drinking water volume,food intake amount,body length and tail length were re...

Abstract:Non-human primates is closer to human than rodents in phylogenetics so that they could be more compatible to simulate clinical symptoms and the mechanism.In this article,we looked ahead at the prospect of non-human primates models,especially the advantage of the use of tree shrews in focal cerebral ischemia research area through reviewing the methods and application of focal cerebral ischemia models in non-human primates at home and abroad.

Abstract:Type 2 diabetes is a complex and heterogeneous disorder presently affecting more than 200 million people worldwide and causing serious socio-economic problems.Appropriate experimental models are essential tools for understanding the pathogenesis,complications,and genetic or environmental effects that increase the risks of diabetes,and inventing various therapeutic agents.The animal models of diabetes can be obtained either spontaneously or induced by chemicals,dietary or surgical manipulations and/or by com...