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7 d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.     

表现为厌食和胰腺炎的胰腺胃重复囊肿

患者 ,女 ,17岁 ,2年前出现间歇性上腹痛并向腰背部放射 ,合并有厌食。腹部超声和 CT显示右侧附件肿块并游离腹腔积液。行诊断性剖腹术时发现右侧卵巢 Morgagni囊肿 ,5 cm大小 ,即行引流术。患者术后上腹部痛依旧且伴进食加剧、体重减轻 ,继而诊断为消化性溃疡、幽门螺杆菌性胃炎、紧张焦虑性肠激惹症 ,针对后者予抗抑郁治疗。患者又由于脱水收住院治疗 ,但仍然腹痛、恶心、呕吐 ,进一步体重减轻。当体重由初诊时 99磅减至 77磅时 ,患者又因厌食及食欲过盛被收住精神科治疗。 1999年 2月 ,患者经历了 6年严重呕吐和腹痛后被体检发现左上腹…     

显微内镜椎间盘摘除术治疗老年腰椎间盘突出症

[目的]探讨老年椎问盘突出症临床特点,评价显微内镜椎间盘摘除术治疗老年腰椎问盘突出症目的疗效和安全性.[方法]对21例诊断为腰椎间盘突出症目的老年患者目的临床特点、影像学资料、显微内镜椎间盘摘除术治疗效果进行分析总结.[结果]所有患者年龄在60岁以上,腰腿痛为主要症状,病史中均无间歇性跛行,21例患者中,L5S1椎间盘突出 2例,L4、5间隙13例,L3、4间隙6例.所有患者行显微内镜椎间盘摘除术,术后恢复优良率90%,平均16个月,随访优良率80%.[结论]老年椎间盘突出症指征掌握适当,显微内镜椎间盘髓核摘除手术可取得优良目的疗效,椎间融合术不应作为常规目的手术选择.     

脑明治疗高血压性脑出血40例疗效观察

我科在 1999年 3月～ 2 0 0 1年 2月 ,用脑明治疗 40例高血压性脑出血进行临床观察 ,现总结如下 :1　临床资料1 1　入选条件　 (1) 5 0～ 80岁 ;(2 )发病 72ｈ内 ;(3 )出血量 <3 0ｍｌ;(4)为高血压引起的脑出血 ;(5 )未破入脑室或蛛网膜下腔者 ;(6)意识清醒或轻度嗜睡能唤醒者。1 2　一般资料　全组病例均选自我科 1999年 3月～ 2 0 0 1年 2月 80例高血压性脑出血完整病例。均经头颅ＣＴ证实 ,符合1995年全国脑血管病学术会议统一诊断标准。随机分成治疗组 40例 ,男 2 4例 ,女 16例 ,平均 5 8 80± 11 0 0岁 ,出血量 19 48± 3 67ｍｌ。对…     

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.     

尤瑞克林联合银杏达莫注射液治疗进展性脑梗死

目的探讨尤瑞克林联合银杏达莫注射液治疗进展性脑梗死的临床效果。方法将80例患者随机分为观察组和对照组，每组40例，两组患者均根据病情给予对症处理。对照组给予银杏达莫注射液20ml加入生理盐水注射液250ml，静脉滴注，每天1次，连续应用15d。观察组在对照组治疗基础上给予尤瑞克林0．15PNA U加入氯化钠注射液100ml，静脉滴注，前10min缓慢滴注，30rain内滴注完毕，每天1次，连续应用7d。患者治疗前和治疗后清晨采集空腹肘静脉血，检查红细胞变形指数和超敏c反应蛋白水平。结果两组患者治疗前后神经功能缺损评分、红细胞变形指数和超敏c反应蛋白检测结果比较，差异有统计学意义（P〈0．05）。观察组总有效率为95．0％，对照组总有效率为80．0％，两组比较差异有统计学意义（P〈0．05）。结论尤瑞克林联合银杏达莫注射液能够显著改善进展性脑梗死患者神经功能缺损状况，减轻脑梗死炎症反应，提高红细胞变形能力，临床治疗效果显著，值得临床推广应用。     

同种异体肌腱联合股前外侧皮瓣一期修复手背软组织缺损

目的 评价同种异体肌腱加皮瓣对手背复合软组织缺损一期修复重建的临床疗效. 方法 2006年7月至2011年7月,对15例手背复合软组织缺损患者一期采用股前外侧皮瓣联合同种异体肌腱行手背伸肌腱修复和创面覆盖,皮瓣大小9 cm×5 cm ～ 14 cm×11 cm,每例修复手背肌腱缺损2～4条,术后2周皮瓣成活后利用被动伸指支具进行早期康复训练. 结果 15例皮瓣均成活,12例患者术后获得随访12 ～ 24个月,平均16个月.2例患者因肌腱粘连术后6个月行肌腱松解,其余10例患者手指屈伸良好.随访结束时,患手腕关节主动屈40°～ 70°,伸25°～50°,掌指关节60°～85°、指间关节80°～90°活动范围,总体优良率达92％. 结论 一期同种异体肌腱联合皮瓣修复手背皮肤及伸指肌腱缺损的方法安全可靠,疗效肯定.同期异体肌腱重建不仅可以避免自体肌腱移植引起的新创伤,而且可以确保及时的手指康复训练,避免了延期手术所致的伸指功能丢失.     

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.