新生大鼠骨骼肌卫星细胞的原代培养及标记方法

本文用等密度沉降离心法分离出新生大鼠骨骼肌卫星细胞,观察其生长规律,检测分裂指数及生长曲线。用胶体金、DAPI及Brdu标记液分别标记细胞8、24、48h,测标记后细胞的生长曲线及标记率。结果表明分离所得肌卫星细胞纯度95％以上,梭形,培养第5天细胞排列出现方向性,并开始融合成肌管,第3—5天增殖旺盛,5代以内生长较快。三种标记方法对肌卫星细胞有不同程度毒性。胶体金标记法毒性最小,标记率100％;DAPI标记细胞率为100％;Brdu毒性最大且标记率仅为10％。本实验方法简单、快速,所得肌卫星细胞纯度高,性能好。胶体金标记法适合于电镜研究,DAPI标记法适合于光镜研究,Brdu标记法较差。     

细胞移植治疗心梗中移植细胞的标记与追踪

成熟个体的心肌细胞属于终未分化细胞，一旦损伤只能由疤痕组织代替。溶栓、冠状动脉血管成型术等治疗手段可以减少梗死面积，保护心功能，提高患者生存率，但却不能逆转已坏死的心肌。而心肌细胞数量减少及心肌重构导致的心衰就成为患者的主要死因。目前的供体细胞主要有胎期心肌细胞、骨骼肌卫星细胞，这些移植细胞能够在宿主心肌内生长，增加了心肌细胞数量，加强了疤痕组织的弹性，限制了疤痕组织的扩     

百福生胶囊对恶性肿瘤患者免疫功能的临床评价

目的 观察百福生胶囊对恶性肿瘤患者免疫功能的临床疗效。方法 纳入临床观察的67例恶性肿瘤患者随机分为实验组与对照组,实验组口服百福生胶囊,对照组口服薏参胶囊,连续治疗1个月后评定疗效。结果 百福生胶囊能显著提高外周血CD3、CD4、CD4/CD8比值及NK细胞(P<0.05),并可明显升高患者血清中的IL-2、IFN-r、TNF水平,降低sIL-2R水平(P<0.01),从而抑制和杀伤肿瘤细胞。结论 百福生胶囊能显著提高恶性肿瘤患者的细胞免疫功能,增强机体免疫力,改善临床症状,提高生存质量。     

大鼠骨骼肌成肌细胞与乳鼠心肌细胞共培养对缝隙连接蛋白43表达的影响

目的 观察体外共培养的乳鼠心肌细胞与大鼠骨骼肌成肌细胞(简称L6细胞)以及乳鼠心肌细胞与转染外源性缝隙连接蛋白43(Cx43)基因的L6细胞(简称L6-Cx43细胞)间Cx43蛋白的表达情况,从而探讨共培养时心肌环境对成肌细胞的作用.方法 将乳鼠心肌细胞与4′,6-二脒基-2-苯基吲哚标记的大鼠骨骼肌成肌细胞进行共培养...     

邵经明教授临证用穴规律探讨

邵经明教授在精研医经、广撷博采、勤于实践的基础上形成了一套独特的学术思想和技术专长,在几十年的临床工作中,以中医理论为基础,以经络学说为指导,遵循辨证施治规律,临证针灸治病,强调"取穴有主次,施术有先后",对临床各科病症的治疗,常以3穴为主而应用.其选穴精当,力专效宏.     

雌、孕激素对LPA3在小鼠子宫内膜围着床期表达的影响

目的 研究小鼠围着床期子宫内膜溶血磷脂酸受体3（LPA3）的表达以及雌、孕激素和孕激素拮抗剂RU486对其表达的影响。方法 用免疫组化法,检测妊娠第1~6天（d1~6）小鼠子宫内膜LPA3蛋白的表达规律; 用RT-PCR、Western blot检测子宫内膜LPA3的表达。结果 妊娠第1~6天小鼠子宫内膜组织均有LPA3表达,d3表达开始增强,d4最强,d5和d6 表达骤然下降。去势小鼠子宫内膜表达低水平LPA3 ;孕激素提高子宫内膜LPA3 的表达,且呈时间依赖性。雌激素对LPA3 表达无明显影响。雌、孕激素联合应用,雌激素降低孕激素对子宫内膜LPA3 表达的上调作用。RU486减弱孕激素的上调作用。结论　LPA3可能参与了胚胎的种植。雌、孕激素可能通过影响小鼠子宫内膜LPA3 的表达,共同参与小鼠子宫内膜容受性的建立。     

邵经明教授"五针法"治疗哮病的方法简介

哮病是一种常见的反复发作、不易根治的慢性顽固性疾病,中医认为,其基本病理变化为“伏痰”遇感引触,痰随气升,气因痰阻,相互搏结,壅塞气道,肺管挛急狭窄,通畅不利,肺气宜降失常,引动停积之痰,而致痰鸣如吼,气息喘促。初病者,其病在肺;病程日久,反复发作,则病及     

大鼠骨髓间充质干细胞移植治疗心肌梗死的实验研究

目的　研究骨髓间充质干细胞(BMSCs)移植对心肌梗死大鼠左心功能的影响以及在心肌病大鼠体内存活、分化的情况。 方法　结扎左冠状动脉前降支制备大鼠心肌梗死模型,采用同种异体大鼠BMSCs 体外分离、纯化、扩增,胶体金标记。按照对BMSCs的处理方式和心肌内注射的成分不同随机分为3组:A组接受BMSCs移植治疗;B组移植经5-氮胞苷体外诱导的BMSCs;C组列为对照组,注射等量培养基;每组12只。4周后通过血流动力学各项指标评价BMSCs移植对大鼠心功能的改善情况;免疫组织化学法检测移植细胞的存活和分化。 结果 与对照组C组相比,移植组A, B两组左心功能显著改善（P<0.01）。A, B两组之间无显著性差异。移植组左心室收缩压（LVSP）、左心室等容期压力最大变化速率（±dp/dtmax）明显高于对照组（P<0.01）,左心室舒张末期压（LVEDP）明显低于对照组（P<0.01）。移植的BMSCs能在心外膜下、梗死区及与正常心肌的交界处存活并向心肌细胞分化, 心肌特异性肌钙蛋白T（cTnT）和α－横纹肌肌动蛋白（α-sarcomeric actin）表达阳性。 结论 同种异体BMSCs移植入大鼠心肌梗死区后能明显改善心功能并向心肌细胞转化。     

碱性成纤维细胞生长因子体外诱导大鼠骨髓间充质干细胞分化为心肌样细胞

Objective To study the feasibility of basic fibroblast growth factor （bFGF）in inducing bone marrow mesenchymal stem cells(BMSCs) to differentiate into cardiomyocyte-like cells in vitro. Methods SD rat BMSCs were isolated and cultured from rat bone marrow, and then induced by bFGF. The cultured cells were observed by a phase-contrast microscope. The immunohistochemical technique and laser scanning confocal microscope (LSCM) were used for examination of the expression of desmin, α-actin and C-TnT. The ultrastructure of induced cells was observed by a transmission electron microscope. GATA4 andα-MHC expressions were detected by relative quantitative RT-PCR after 7, 21, 28 days of induction respectively. Results After primary cells were cultured for 48 hours,most of them became fusiform fibroblast-like shape with two or three processes and a few of them were flat in shape. The morphology of BMSCs induced by bFGF changed obviously. After being induced by bFGF for one week, the cells became larger and most of them became myocyte-like short column in shape or long shuttle-shape while paralleled. After four weeks, most cells became short column-shape and touched with adjacent cells tightly to form myotube-like structure,with apparent directionality of cell arraying. BMSCs induced by bFGF were identified by the positive staining for desmin, α-sarcomeric actin and C-TnT. Of BMSCs, there were more desmin positive and α-actin-positive cells made up higher of all BMSCs than C-TnT-positive cells. Desmin and α-actin-positive cells were about 33.82% and 58.64%, and C-TnT-positive cells were about 28.94%. Desmin(α-actin ) appeared red, and C-TnT was green under a laser scaning confocal microscopy. Their co-expression appeared yellow. Transmission electron microscope showed that the induced cells were rod in shape and the ovoid nuclei were positioned in the center of the cell. lots of mitochondria, rough endoplasmic reticulum, ribosome and paralleled myofilaments were founded in plasm.RT-PCR assessment showed that the differentiated cells began to express GATA-4 from day 7 to day 28 of differentiation and began to express α-myosin heavy china(α-MHC) fro     

催产素体外诱导大鼠骨髓间充质干细胞分化为心肌样细胞

目的 应用催产素(OT)体外诱导骨髓间充质干细胞(BMSCs)向心肌样细胞分化,探索BMSCs向心肌细胞分化的诱导方法.方法 取SD大鼠四肢骨骨髓,分离培养BMSCs,应用OT定向诱导,相差显微镜观察细胞形态学变化,应用免疫细胞化学、激光扫描共焦显微镜技术检测结蛋白、α-横纹肌肌动蛋白、心肌特异性肌钙蛋白T(C-TnT...