Study on chondrogenic differentiation of canine mesenchymal stem cells induced by Type 2 recombinant adeno-associated viral mediated transfer of hTGF-β1

Objective To investigate the potential application of human transforming growth factor-beta-1 (hTGF-β1) gene mediated by type 2 recombinant adeno-associated virus (rAAV2) vector inducing chondrogenic differentiation of canine mesenchymal stem cells (MSCs) in vitro. Methods Canine MSCs from bone marrow were isolated and cultured in vitro by density gradient centrifngation and adherence screening methods. The morphology of MSCs was observed by inverted phase contrast microscope and Giemsa stain. Flow eytometry was used to detect surface antigens of MSCs, The third generation of MSCs were transfected by rAAV2-hTGF-β1 with or without MOI of 1 ×105 v.g./cell or 5×105 v.g./cell. The expression of hTGF-β1 was detected by Western blot after 10 days, and TGF-β1 synthesis was determined by ELISA at 3, 6 and 9 day, respectively. After 2 weeks of culturing, mRNA expressions of type Ⅱ collagen and aggrecan were determined by RT-PCR and the collagen Ⅱ protein was detected by immunocytochemistry. Results The MSCs appeared to be morphologically spindle-shaped and showed active capability of proliferation both in primary and passage generations. Flow cytometry analysis indicated that MSCs were universally positive for CD29, CD44 and CD105, but negative for CD34 and CD45. TGF-β1 expression can be observed by Western blot after 10 days in two transfection groups, MOI of 5 × 105 group and MOI of 1× 105 group. With the extension of time, the contents of hTGF-β1 increased in the two groups detected by ELISA, while there was a significant difference between them two (P < 0.01). After 2 weeks of transfection of MSCs by rAAV2-hTGF-β1, the expression of collagen Ⅱ and Aggreacan mRNAs were positive. It also showed positive of collagen Ⅱ detected by immunocytochemistry. Conclusion Canine MSCs show chondrogenesis differentiation after induction by Type 2 rAAV mediated transfer of TGF-β1 gene. The process is a potential application for cartilage tissue engineering.     

低分子肝素钙治疗进展性脑梗死疗效观察

目的 探讨低分子肝素钙在治疗进展性脑梗死的疗效观察.方法 将120例进展性脑梗死住院患者,随机分为观察组60例与对照组60例.两组应用常规治疗,观察组加用低分子肝素钙.结果 观察组有效率明显高于对照组.结论 进展性脑梗死常规治疗联合低分子肝素钙治疗,比单纯常规治疗,在预防脑梗死家中效果显著,安全性高,值得推广.     

丹参内关穴封闭缓解急性心肌梗死

急性心肌梗死 (ＡＭＩ)出现剧烈的胸痛 ,在应用一般镇痛措施效差的情况下 ,采取丹参内关穴封闭的方法获得满意的疗效 ,报告如下 :1　临床资料选择临床上符合ＷＨＯ诊断标准的ＡＭＩ患者 18例。其中急性前壁心肌梗死 12例 ,广泛前壁心肌梗死 6例 ;年龄 5 0～ 76岁 ,无高血压病史。全部病例均采用了杜冷丁或吗啡肌肉或静脉注射 ,硝酸甘油舌下含化或静脉滴注及其它镇静止痛药物 ,部分病例应用了静脉溶栓 ,但患者仍出现剧烈胸痛。即应用 5ｍｌ注射器抽取复方丹参 (上海第一制药厂生产 ) 2ｍｌ,左侧内关穴局部消毒后 ,采用直刺方法 ,反复提插几次…     

高血压性脑出血并神经源性肺水肿的临床与CT表现

我院在1998年1月至2003年3月共收治高血压性脑出血788例，其中36例合并神经源性肺水肿（NPE）。本文对36例高血压性脑出血并NPE的临床及CT表现，现分析如下：     

适脑脉治疗脑梗死患者运动及认知功能的临床观察

观察适脑脉对脑梗死患者运动及认知功能有明显疗效 ,现报道如下 :1　材料与方法1 1　病例选择　 1998年 1月至 2 0 0 1年 4月在内科住院的 5 6例脑梗死恢复期患者 ,男 3 2例 ,女 2 4例 ,年龄 40～ 78岁 (平均5 6 42± 12 40岁 ) ,符合 1986年全国第二次脑血管会议通过的诊断标准 ,并经ＣＴ检查证实。入院治疗时意识清醒 ,梗塞部位 :基底节区 2 8例 ,放射冠 16例 ,额叶 6例 ,颞、枕叶各 3例。病程 8天～半年 ,平均 3 6月。1 2　治疗方法1 2 1　药物治疗 :常规组应用血管扩张剂及其它对症处理。适脑脉组是在常规治疗的基础上给予适脑脉 (瑞…     

甲状腺功能亢进性心脏病27例误诊分析

甲状腺功能亢进症 (以下简称甲亢 )是临床常见的一种内分泌疾病 ,并发症较多 ,其中以甲状腺功能亢进性心脏病 (以下简称甲亢心脏病 )最常见 ,因其心脏症状表现突出 ,特别当甲亢症状不典型时 ,容易造成误诊。我院 1994年 6月— 1998年 10月共收治 2 7例甲亢心脏病误诊病例 ,现将误诊原因分析如下。1　临床资料1.1一般资料　男 10例 ,女 17例 ,年龄 8～ 6 8岁 ,平均 42岁 ,其中 2 2例为门诊患者 ,5例为住院患者 ,院外误诊 17例 ,院内误诊 10例 ,首次就诊至确诊时间 1～ 36个月 ,平均 6个月。1.2临床表现　 2 7例患者均有心慌、胸闷、心率增快 …     

TGF-β1壳聚糖缓释微球的制备和体外检测

【目的】通过制备新型的转化生长因子-β1(TGF-β1)缓释系统，并对其载药、体外释药及降解特性进行检测，评估应用生物可降解壳聚糖微球作为TGF-β1控制释放载体的可行性。【方法】应用三聚磷酸钠(TPP)作为交联剂，以乳化交联法制备具有控制释放功能的负载TGF-β1的壳聚糖微球；以牛血清白蛋白(BSA)作为模板蛋白，应用相同的方法制备负载。BSA的微球。应用扫描电镜、微镜粒度分、药物包封率测定、体外药物释放动力学检测等方法分析微球的特性。【结果】制备的微球球形良好，球体表面光滑，具有较高的TGF-β1包封效率90．1％)。持续7d的药物释放试验表明，BSA与TGF-β1两种蛋白均可以从微球中缓慢释放，其中TGF-β1的释放率低于BSA的释放率。溶菌酶溶液降解作用下，4周的体外降解过程中，可见微球质量持续下降并伴有明显的微球形貌改变。【结论】应用乳化交联法可制备负载TGF-β1的壳聚糖缓释微球。这种新型药物控制释放系统在细胞因子的控制释放及软骨组织工程中有潜在的应用价值。     

侧脑室穿刺引流并腰穿置换脑脊液治疗脑室出血46例

我科自 1998年 4月至 2 0 0 1年 4月 ,对 46例脑室出血(IVH)患者采用侧脑室穿刺加尿激酶 (UK)溶解并腰穿置换脑脊液治疗 ,与 1993年 4月至 1998年 3月的 48例采用侧脑室穿刺加 U K溶解引流的 IVH患者进行对照研究 ,效果满意 ,现报道如下。1　资料与方法1.1　本研究中 94例均为我院住院患者 ,均经 CT所证实的原发性脑室出血 (PIVH )或以脑室积血为主的继发性脑室出血 (SIVH)。1.1.1　治疗组　 46例 ,男 2 9例 ,女 17例。年龄 18～ 80岁 ,平均 5 7岁。PIVH11例 ,SIVH35例。单侧侧脑室积血 7例 ,双侧脑室并三脑室积血 2 3例 ,单纯…     

Study on chondrogenic differentiation of canine mesenchymal stem cells induced by Type 2 recombinant adeno-associated viral mediated transfer of hTGF-β1

Objective To investigate the potential application of human transforming growth factor-beta-1 (hTGF-β1) gene mediated by type 2 recombinant adeno-associated virus (rAAV2) vector inducing chondrogenic differentiation of canine mesenchymal stem cells (MSCs) in vitro. Methods Canine MSCs from bone marrow were isolated and cultured in vitro by density gradient centrifngation and adherence screening methods. The morphology of MSCs was observed by inverted phase contrast microscope and Giemsa stain. Flow eytometry was used to detect surface antigens of MSCs, The third generation of MSCs were transfected by rAAV2-hTGF-β1 with or without MOI of 1 ×105 v.g./cell or 5×105 v.g./cell. The expression of hTGF-β1 was detected by Western blot after 10 days, and TGF-β1 synthesis was determined by ELISA at 3, 6 and 9 day, respectively. After 2 weeks of culturing, mRNA expressions of type Ⅱ collagen and aggrecan were determined by RT-PCR and the collagen Ⅱ protein was detected by immunocytochemistry. Results The MSCs appeared to be morphologically spindle-shaped and showed active capability of proliferation both in primary and passage generations. Flow cytometry analysis indicated that MSCs were universally positive for CD29, CD44 and CD105, but negative for CD34 and CD45. TGF-β1 expression can be observed by Western blot after 10 days in two transfection groups, MOI of 5 × 105 group and MOI of 1× 105 group. With the extension of time, the contents of hTGF-β1 increased in the two groups detected by ELISA, while there was a significant difference between them two (P < 0.01). After 2 weeks of transfection of MSCs by rAAV2-hTGF-β1, the expression of collagen Ⅱ and Aggreacan mRNAs were positive. It also showed positive of collagen Ⅱ detected by immunocytochemistry. Conclusion Canine MSCs show chondrogenesis differentiation after induction by Type 2 rAAV mediated transfer of TGF-β1 gene. The process is a potential application for cartilage tissue engineering.     

罗哌卡因局部浸润用于显微内镜腰椎间盘切除术后镇痛

目的：评价罗哌卡因切口周围局部浸润在显微内镜腰椎间盘髓核摘除术后镇痛中的作用。方法：83例行显微内镜腰椎间盘髓核摘除术患者随机分成2组，A组（n=43）手术开始前以0．75％盐酸罗哌卡因10mL于切口周围肌层和皮下局部浸润；B组（n=40）手术开始前以0．9％生理盐水10mL于切口周围肌层和皮下局部浸润。对手术后患者的疼痛指数、阿片类镇痛药物使用量以及切口愈合情况等进行统计观察。结果：A组患者的结果明显优于B组，A组患者术后疼痛反应轻、阿片类镇痛药物使用量少、胃肠道反应发生率低、住院时间更短。所有患者未出现切口愈合不良等并发症。结论：罗哌卡因切1：2周围局部浸润对于缓解显微内镜腰椎间盘髓核摘除术后疼痛具有优良的效果，能明显缩短患者术后恢复的时间，值得临床推广使用。