电磁脉冲诱导大鼠心肌闰盘Cx43连接蛋白的上调

目的:探讨电磁脉冲(EMP)对SD大鼠心肌闰盘及主要连接蛋白Cx43的影响.方法:将雄性SD大鼠35只,随机分为对照组和EMP辐照组,EMP辐照组又分为照后即刻、1h、3h、6h、12h、24h组,每组5只大鼠.应用EMP模拟发生器对EMP辐照组大鼠进行辐照,场强为200 kV/m、前沿15 ns、脉宽7-8 ns、脉冲次数200次,脉冲间隔为7s,麻醉后取大鼠左心室,采用硝酸镧示踪法和透射电子显微镜观察大鼠心肌闰盘(ID)结构的变化,用Real-Time PCR方法和Western blot方法检测EMP辐照后,大鼠连接蛋白Cx43在转录水平和蛋白水平表达量的变化情况.结果:电镜下正常对照组大鼠心肌ID处未见硝酸镧颗粒沉积;EMP组大鼠ID处可见硝酸镧颗粒,且随时间的延长,硝酸镧颗粒沉积量逐渐增多,以照后6h组硝酸镧颗粒沉积量达最大,然后硝酸镧颗粒沉积量随时间延长逐渐减少,24h组恢复正常.心肌细胞Cx43的mRNA表达水平在EMP照后lh和6h明显增高,分别是对照组的1.95、3.10倍(P＜0.05);照后即刻、3h、12h、24h表达水平与对照组相比差异无统计学意义(P＞0.05).Western blot结果为照后即刻、1h、3h和6h组大鼠心肌细胞Cx43蛋白表达量与对照组比较显著增加,12h、24h与对照组相比差异无统计学意义(P＞0.05).结论:EMP可使心肌闰盘结构发生改变,上调Cx43蛋白的mRNA水平和蛋白表达量.     

发育心肌和心肌干细胞分化后心肌特异蛋白表达的比较研究

为探讨MCSC移植修复缺血性心肌的可能性,观察了大鼠发育心肌和MCSC分化为心肌细胞的cTnT和Cx-43表达特点。结果表明从胚胎发育至成年,心肌cTnT mRNA表达逐渐上调;用BMP-2诱导1周的MCSC可检测到cTnT mRNA表达,3～4周表达显著升高,诱导2周能观察到cTnT蛋白,3～4周可见cTnT呈现横纹样结构。胚胎第11 d心肌可检测到Cx-43 mRNA表达,生后7 d达高峰,以后逐渐降低,17 d趋于稳定。胚胎期Cx-43蛋白多分布于肌膜下,生后10 d位于细胞连接处。诱导的MCSC中Cx-43 mRNA表达特征与cTnT mRNA相似,2周可观察到Cx-43蛋白,3～4周可见Cx-43位于相邻细胞连接处。本研究结果提示,在BMP-2诱导下MCSC可分化为心肌细胞,表现为成熟心肌细胞的结构特征。MCSC有望成为治疗缺血性心脏病的理想种子细胞。     

心肌肥厚大鼠心肌Cx43的表达及法舒地尔的干预作用

目的：研究腹主动脉缩窄大鼠心肌缝隙连接蛋白Cx43的变化及法舒地尔的干预作用。方法：腹主动脉缩窄建立心肌肥厚大鼠模型,随机分假手术组,腹主动脉缩窄组、腹主动脉缩窄＋10mg／kg法舒地尔（ip,每天1次,8周）组、腹主动脉缩窄＋40mg／kg（ip,每天1次,8周）。病理切片观察心肌组织学变化;免疫组化法检测心肌Cx43蛋白表达。结果：模型组大鼠心肌细胞排列紊乱,肥大,间隙增宽,Cx43蛋白表达量明显低于正常组;Fas治疗后,死亡率下降,cx43蛋白表达量高于模型组,差异具有统计学意义（P〈0．01）。结论：Fas可能通过调高Cx43蛋白表达,治疗大鼠心肌肥厚。     

成年牦牛与双性牦牛睾丸蛋白质双向电泳的比较研究

为了比较牦牛和双性牦牛睾丸组织中蛋白质的差异表达,以探索双性牦牛生殖障碍的机理。提取牦牛睾丸(n=4)和1头双性牦牛睾丸总蛋白质,采用双向电泳分离后,对差异表达蛋白质点进行质谱鉴定。实验获得了分辨率高、重复性好的牦牛与双性牦牛睾丸组织蛋白质的双向电泳图谱,2倍及以上差异表达的蛋白质24个,其中22个蛋白质经质谱分析和数据库搜索后得到鉴定。有6种蛋白质属于分子伴侣,在双性牦牛睾丸组织中表达量显著下降,推测与双性牦牛精子发生障碍有关。     

低频脉冲磁场对大鼠心肌微血管内皮细胞增殖和缝隙连接蛋白43表达的影响

目的:探讨低频磁场对大鼠心肌微血管内皮细胞(CMECs)增殖和缝隙连接蛋白43(Cx43)表达的影响.方法:设不同照射强度为(08.mT组,1.4mT组,1.8mT组)为照射组.不加磁场干预为对照组.照射条件;磁场频率为15Hz,强度分别为0.8mT、1.4mT、1.8mT,照射时间为4 h/d,连续照射7d.应用MTT法检测CMECs增殖,采用Western blot检测Cx43蛋白表达.结果:生长曲线结果显示,磁场能够促进CMECs增殖.1.4mT组照射第2d后CMECs生长速度加快,在第3d开始进入对数生长期,在第4d生长最为旺盛之后进入生长平台期,第2～7d与对照组比较有显著性差异(P＜0.05),而且细胞生长曲线明显前移并且峰值增高.1.4mT组、1.8mT组CMECs增殖与对照组比较显著升高(P＞0.05).磁场照射后Cx43表达明显上调,1.4mT组、1.8mT组Cx43蛋白的表达均明显上升,与对照组比较差异有统计学意义(P＜0.05),0.8mT组cx43蛋白的表达与对照组比较无显著性差异(P＞0.05).而Cx43蛋白的表达1.4 mT组和1.8mT组之间差异无统计学意义(P＞0.05).结论:低频脉冲磁场能促进CMECs增殖与增强细胞活力,上调Cx43表达,其在分子水平上的可能作用机制表现为对Cx43的有效调控.     

天祝白牦牛与黄牛NGB蛋白三级结构比较分析

通过比较天祝白牦牛(Bos grunniens)与黄牛(Bos taurus)NGB蛋白三级结构,分析天祝白牦牛NGB蛋白氨基酸位点突变对其三级结构的影响.利用生物信息学在线分析网站及软件对天祝白牦牛和黄牛NGB蛋白的同源性、系统发育进化、三级结构进行比较分析.结果显示,天祝白牦牛NGB蛋白进化趋于保守,83号位点为未...     

低氧对大鼠右心室肥厚及心肌中缝隙连接蛋白43表达的影响

目的:研究低氧对大鼠右心室肥厚及心肌中缝隙连接蛋白43 (Cx43)表达的影响.方法:40只健康雄性SD大鼠随机分为正常组(control)、低氧3周组、低氧4周组和低氧5周组.除正常组外,其余3组大鼠分别在低氧环境中饲养3周、4周和5周.测定和比较各组大鼠的平均肺动脉压力(mPAP)、右心室收缩压(RVSP)、右心室肥厚度[Rv/(LV+S)％],并通过免疫组化染色法观察各组大鼠左心室心肌细胞中cx43的表达.结果:与正常组相比,低氧3周、4周、5周组大鼠的mPAP、RVSP、右心室肥厚度均显著升高(P均＜0.05),与低氧3周组比较,低氧4周、5周组大鼠的mPAP、RVSP、右心室肥厚度均显著升高(P均＜0.05),而低氧5周组大鼠的mPAP、RVSP、右心室肥厚度均显著高于低氧4周(P均＜0.05).免疫组织化学结果显示:低氧组大鼠Cx43排列紊乱,端-端连接减少,侧面连接增多;随着低氧时间的延长,大鼠心肌细胞中Cx43的表达逐渐减少,差异具有统计学意义(P均＜0.05).结论:低氧可导致右心室肥厚,并随着诱导时间的延长而逐渐加重,这可能与心肌中Cx43的分布紊乱及表达减少有关.     

心肌型脂肪酸结合蛋白胶体金检测试纸条的制备和初步应用

目的应用胶体金免疫层析法制备检测全血或血清样本中心肌型脂肪酸结合蛋白（H-FABP）的检测试纸条,用于急性心肌梗塞（AMI）的早期辅助诊断。方法采用柠檬酸三钠还原法制备胶体金,标记鼠抗心肌型脂肪酸结合蛋白单克隆抗体,喷于玻璃纤维膜上制成胶体金结合物垫,将另一株鼠抗心肌型脂肪酸结合蛋白单克隆抗体和抗鼠二抗分别包被检测线和质控线,组装成试纸条进行灵敏性、特异性、精密性、稳定性及临床样品检测。结果该试纸条的检测灵敏度为10ng／mL,15min内可判定结果;与肌钙蛋白I、C反应蛋白、肌酸激酶、人心肌肌红蛋白无交叉反应。检测240份临床标本,与临床诊断结果进行配对分析,阳性符合率95．83％、阴性符合率100％、总符合率97．92％。结论制备的H-FABP检测试纸条有良好的灵敏性、特异性,可用于早期AMI的辅助诊断。     

贯叶连翘提取物对扩张型心肌病大鼠缝隙连接蛋白Cx43的影响

目的:研究贯叶连翘提取物(HPE)对扩张型心肌病(DCM)大鼠心肌缝隙连接蛋白Cx43表达的作用。方法:以腹腔注射阿霉素建立DCM大鼠模型为基础,分析心肌缝隙连接蛋白(Cx43)表达差异性。结果:治疗组大鼠Cx43蛋白以及Cx43mRNA显著高于模型组。结论:HPE可能通过对Cx43受体敏感性的调节而改变Cx43表达,改善DCM大鼠心功能。     

稳心颗粒联合盐酸决奈达隆片对阵发性房颤患者氧化应激、心肌纤维化和外周血单核细胞Cx40、Cx43的影响

摘要 目的：观察稳心颗粒联合盐酸决奈达隆片对阵发性房颤（PAF）患者氧化应激、心肌纤维化和外周血单核细胞缝隙连接蛋白40（Cx40）信使核糖核酸（mRNA）、缝隙连接蛋白43（Cx43）mRNA的影响。方法：选取2020年3月~2022年12月期间在我院治疗的79例PAF患者为研究对象。按照随机数字表法将患者分为对照组（39例,盐酸决奈达隆片治疗）和研究组（40例,稳心颗粒联合盐酸决奈达隆片治疗）。对比两组房颤（AF）发作次数、静息心率、AF持续时间、氧化应激指标、心肌纤维化指标和外周血单核细胞Cx40mRNA、Cx43mRNA的变化情况。结果：与对照组治疗12周后相比,研究组的AF发作次数更少,静息心率更低,AF持续时间更短,丙二醛（MDA）、Ⅲ型前胶原氨基末端肽（PⅢNP）、基质金属蛋白酶-2（MMP-2）、外周血单核细胞Cx40mRNA、Cx43mRNA更低,超氧化物歧化酶（SOD）更高（P<0.05）。结论：稳心颗粒联合盐酸决奈达隆片治疗PAF患者,可有效改善患者的临床症状,减轻患者的氧化应激、心肌纤维化损伤,调节外周血单核细胞Cx40mRNA、Cx43mRNA水平。     

牦牛海马的形态特征及成体神经发生的初探

采用传统H.E 染色和Golgi-Cox 染色方法观察成年牦牛海马结构的形态和细胞构筑,并通过DCX - DAB免疫组化染色和DCX/ NeuN、GFAP / NeuN 双重免疫荧光标记等技术观察齿状回颗粒下层中的新生神经元和放射状胶质细胞。结果表明,牦牛海马结构主要包括齿状回和海马本部,二者分层清晰。海马的主要细胞为颗粒细胞、苔藓细胞和锥体细胞。CA3 区的锥体细胞胞体较CA1 区的大,但其顶树突的平均长度较短。CA1 区的锥体细胞明显分为两层,而CA3 区的则为一层。DCX 阳性细胞的胞体主要集中在齿状回颗粒下层靠近门区处,沿颗粒层内侧单个或少数聚集分布。沿齿状回颗粒下层分布着一层GFAP 阳性的放射状胶质细胞样细胞,其胞质和单极性的细长突起均呈GFAP 阳性,而胞核为阴性。在整个海马结构中均有大量星形GFAP 阳性细胞散在分布,特别是海马分子层和门区内靠近颗粒层部分的密度较其它部位大。牦牛海马的形态结构与绵羊的相似,而与大鼠、小鼠、家猫、兔子等小型哺乳动物有一定差别。两种DCX 免疫组化实验结果表明在牦牛海马中存在着新生神经元。GFAP 免疫荧光标记表明,牦牛海马结构中分布有星形胶质细胞;特别是放射状胶质细胞。     

Morphological study of the atrioventricular conduction system and Purkinje fibers in yak

We studied the morphology of the atrioventricular conduction system (AVCS) and Purkinje fibers of the yak. Light and transmission electron microscopy were used to study the histological features of AVCS. The distributional characteristics of the His‐bundle, the left bundle branch (LBB), right bundle branch (RBB), and Purkinje fiber network of yak hearts were examined using gross dissection, ink injection, and ABS casting. The results showed that the atrioventricular node (AVN) of yak located in the right side of interatrial septum and had a flattened ovoid shape. The AVN of yak is composed of the slender, interweaving cells formed almost entirely of the transitional cells (T‐cells). The His‐bundle extended from the AVN, and split into left LBB and RBB at the crest of the interventricular septum. The LBB descended along the left side of interventricular septum. At approximately the upper 1/3 of the interventricular septum, the LBB typically divided into three branches. The RBB ran under the endocardium of the right side of interventricular septum, and extended to the base of septal papillary muscle, passed into the moderator band, crossed the right ventricular cavity to reach the base of anterior papillary muscle, and divided into four fascicles under the subendocardial layer. The Purkinje fibers in the ventricle formed a complex spatial network. The distributional and cellular component characteristics of the AVCS and Purkinje fibers ensured normal cardiac function.     

A study on the inhibition of rat myocardium glutathione peroxidase and glutathione reductase by moniliformin

The yeasts of patients with oral cancer has been studied before and during Xr-therapy. Gram and PAS smears revealed an increase of yeast-like structures, during treatment, from 56% to 66% of the cases. Before radiotherapy oral yeasts were isolated from 56% of the patients with cancer represented by Candida albicans (30%); C. tropicalis (12%); C. glabrata and C. krusei (4%), besides six other different species (2%). During radiotherapy yeasts were isolated in 72% of the cases, as follow: C. albicans (36%); C. tropicalis (16%); Rhodotorula rubra (6%); C. kefyr; C. krusei and Pichia farinosa (4%), besides other nine species (2%). C. albicans serotype A represented 93% of the isolated samples, before treatment and 88,8% during Xr-therapy.     

在生理温度下降钙素基因相关肽对豚鼠心房肌复极过程的影响

实验应用常规微电极方法研究了在生理温度下 (36 5± 0 5℃ )降钙素基因相关肽 (calcitoningene relatedpeptide ,CGRP)对豚鼠心房肌细胞复极过程的影响及其与钾电流的关系。结果表明 :(1)CGRP(16nmol/L)可拮抗由钾通道阻断剂BaCl2 、4 AP引起的动作电位时间延长。 (2 )CGRP(16nmol/L)能够增加细胞外高钾 (18 5mmol/L)条件下心房肌慢反应动作电位的APA和Vmax,并缩短传导时间。 (3)CGRP(16nmol/L)能减弱甚至消除因并用CsCl (5mmol/L)和无钾灌流液诱发的触发活动。 (4)CGRP对动作电位复极过程的作用因温度条件而异。在生理温度下 ,CGRP(5、16和 5 0nmol/L)能够使动作电位平台抬高 ,缩短动作电位复极化 2 0 %、5 0 %和 90 %时程。其中 ,对动作电位复极化 2 0 %、5 0 %时程的作用呈剂量依赖性。而在室温下 (2 5 5± 2 1℃ ) ,CGRP使动作电位复极化 2 0 %、5 0 %和90 %时程延长。上述结果提示 ,CGRP对心房肌细胞具有多重电生理效应 ,其中生理温度下CGRP对钾电流的促进作用在动作电位的改变中占重要地位 ,今后有必要进一步研究CGRP对各种钾通道的作用     

高原慢性低氧对土生牦牛与迁饲黄牛肾功能影响的比较

目的：探讨长期慢性高原暴露对高原土生牦牛（yak）、高山迁饲黄牛（migratedcattle）和低海拔黄牛（lowland cattle）肾功能的影响,探究牦牛和高山迁饲黄牛适应和习服高原的不同特征及差异。方法：采集青海不同地区的牦牛血样,按海拔高度分为3 000 m,3 500 m,4 000 m和4 300 m四个组（n=84）,同时采集高山迁饲黄牛（n=22）及低海拔黄牛（n=39）血液,利用全自动血液生化分析仪测定尿素氮（BUN）、肌酐（Cr）、尿素氮/肌酐（BUN/Cr）、尿酸（UA）、二氧化碳结合率（CO₂cp）、葡萄糖（GLU）水平,分析不同海拔高度的牦牛之间及牦牛、高山迁饲黄牛和低海拔黄牛之间的差异。结果：随着海拔的升高,土生牦牛UA、CO₂cp两项升高较显著,与黄牛相比,BUN、BUN/Cr显著高于高山迁饲黄牛和低海拔黄牛, CO₂cp、GLU显著低于低海拔黄牛。低海拔黄牛和高山迁饲黄牛相比,BUN、BUN/Cr显著低于高山迁饲黄牛,UA、CO₂cp显著高于高山迁饲黄牛。结论：土生牦牛对于高原低氧环境有很强的适应能力;高山迁饲黄牛表现出对低氧的不适应性,处于对低氧的应激状态。     

Complex formation between calmodulin and a peptide from the intracellular loop of the gap junction protein connexin43: Molecular conformation and energetics of binding

Gap junctions are formed by a family of transmembrane proteins, connexins. Connexin43 is a widely studied member of the family, being ubiquitously expressed in a variety of tissues and a target of a large number of disease mutations. The intracellular loop of connexin43 has been shown to include a calmodulin binding domain, but detailed 3-dimensional data on the structure of the complex are not available. In this study, we used a synthetic peptide from this domain to reveal the conformation of the calmodulin-peptide complex by small angle X-ray scattering. Upon peptide binding, calmodulin lost its dumbbell shape, adopting a more globular conformation. We also studied the energetics of the interaction using calorimetry and computational methods. All our data indicate that calmodulin binds to the peptide from cx43 in the classical ‘collapsed’ conformation.     

Immunoexpression of aquaporins 1, 2, 3 and 4 in kidney of yak (Bos grunniens) on the Qinghai-Tibetan Plateau

Aquaporins (AQP) 1, 2, 3 and 4 belong to the aquaporin water channel family and play an important role in urine concentration by reabsorption of water from renal tubule fluid. Renal AQPs have not been reported in the yak (Bos grunniens), which resides in the Qinghai Tibetan Plateau. We investigated AQPs 1?4 expressions in the kidneys of Yak using immunohistochemical staining. AQP1 was expressed mainly in the basolateral and apical membranes of the proximal tubules and descending thin limb of the loop of Henle. AQP2 was detected in the apical plasma membranes of collecting ducts and distal convoluted tubules. AQP3 was located in the proximal tubule, distal tubule and collecting ducts. AQP4 was located in the collecting ducts, distal straight tubule, glomerular capillaries and peritubular capillaries. The expression pattern of AQPs 1?4 in kidney of yak was different from other species, which possibly is related to kidney function in a high altitude environment.     

A pilot study on the frequency structure of histological neoplasm diagnoses in rats

There are large differences in the frequencies of different kinds of neoplasms in a human or animal population. The question arises, whether the set of these frequencies shows some characteristic feature. Our fitting results on neoplasm frequency data relating to laboratory rats show that the frequencies are approximately lognormally distributed. At the same time, fitting results with the logarithmic series distribution, also frequently used in similar studies, are poor. A good fit of the Zipf-Mandelbrot distribution, fitted to the descendingly ordered dominant and subdominant frequencies can be achieved, sometimes after omitting some diagnoses. We point out the possibility that the omitted frequencies may be considered "unnatural" ones. The good fit of a particular frequency distribution to the diagnosis frequency set suggests a corresponding chance mechanism in forming the occurrence probabilities of different kinds of neoplasms. A frequently used scalar feature of frequency distributions of categorical data is the concentration or diversity. It was found that in the female rat population the diagnoses are more concentrated among the diagnosis categories. A possible explanation may involve the fact that females mature faster than males. The study of the distribution properties of the diagnosis frequencies promises the observation of new epidemiological phenomena.     

Interplay between PKC and the MAP kinase pathway in Connexin43 phosphorylation and inhibition of gap junction intercellular communication

Gap junction channels are made of a family proteins called connexins. The best-studied type of connexin, Connexin43 (Cx43), is phosphorylated at several sites in its C-terminus. The tumor-promoting phorbol ester TPA strongly inhibits Cx43 gap junction channels. In this study we have investigated mechanisms involved in TPA-induced phosphorylation of Cx43 and inhibition of gap junction channels. The data show that TPA-induced inhibition of gap junction intercellular communication (GJIC) is dependent on both PKC and the MAP kinase pathway. The data suggest that PKC-induced activation of MAP kinase partly involves Src-independent trans-activation of the EGF receptor, and that TPA-induced shift in SDS-PAGE gel mobility of Cx43 is caused by MAP kinase phosphorylation, whereas phosphorylation of S368 by PKC does not alter gel migration of Cx43. We also show that TPA, in addition to phosphorylation of S368, also induces phosphorylation of S255 and S262, in a MAP kinase-dependent manner. The data add to our understanding of the molecular mechanisms involved in the interplay between signaling pathways in regulation of GJIC.