含服硝苯吡啶治疗危重症高血压36例疗效观察

我们自 1999年 1月～ 2 0 0 1年 6月 ,用含服硝苯吡啶的方法治疗危重症高血压 3 6例 ,取得满意疗效 ,现报告如下。1　病例与方法1 1　病例　 60例危重症高血压病人 ,按 1998年第七届世界卫生组织 /国际高血压联盟日本会议制订的诊断标准 ,均符合 3级高血压 ,即收缩压≥ 180ｍｍＨｇ ,舒张压≥ 110ｍｍＨｇ ,其中有 3 1例血压超过 2 2 0 /12 0ｍｍＨｇ ,主要症状有头痛、头晕、心悸、胸闷、恶心 ,部分伴有烦躁、呕吐、视物模糊、肢体麻木等。将这些病例随机分为治疗组 3 6例 ,男 2 3例 ,女 13例 ,年龄 41～ 84岁 ,平均 61岁 ,其中高血压危象 …     

透明质酸修饰壳聚糖/pDNA纳米微球的制备及体外转染软骨细胞

目的 以透明质酸(HA)修饰基因载体壳聚糖(CS)纳米微球,制作一种新型的HA/CS/pDNA基因转染系统,观察其结构特征及体外对软骨细胞的转染能力.方法 将不同比例的HA和CS与负载增强型绿色荧光蛋白基因(EGFP)的质粒DNA(pDNA)以复凝聚法制成纳米微球,以扫描电镜检测纳米微球形态,Zeta电位粒度分析仪测定其粒径、表面电位;凝胶电泳阻滞实验观察CS和pDNA的结合力;体外转染兔关节软骨细胞,以流式细胞仪及荧光显微镜检测转染效率.结果 HA/CS/pDNA纳米微球多呈球形,粒径为(223±51)nm,表面Zeta电位为(17.4±6.1)mV,可有效保护pDNA免受 DNaseⅠ的降解.体外转染实验证明HA/CS/pDNA纳米微球能介导pEGFP转染软骨细胞并在细胞内表达绿色荧光蛋白,转染率达(15.450±0.404)%,比裸pDNA组和CS/pDNA组有更高的转染效率(P<0.05).结论 复凝聚法制备的HA/CS/pDNA纳米微球是一种有效的新型非病毒基因转染系统,对软骨细胞有着潜在的靶向基因转染能力.

Abstract：

Objective To prepare hyaluronic acid (HA)-modified chitosan (CS)/pDNA (HA/CS/pDNA) nanoparticles as novel gene vectors, study their structural characteristics and gene transfection efficiency for chondrocytes in vitro in rabbits. Methods The HA/CS/pDNA nanoparticles were prepared by a DNA (pDNA), which load enhanced green fluorescent protein (EGFP) gene. The morphology of the nanoparticles was observed under the transmission electron microscopy. The sizes and zeta-potentials of the nanoparticles were measured by a Marven-nano laser diffractometer. The binding of pDNA was evaluated by agarose gel electrophoresis analysis. The gene transfection experiments in vitro were performed with the chondrocytes of rabbits. The gene transfection efficiency was measured by using flow cytometry and under the fluorescence microscopy. Results HA/CS/pDNA nanoparticles were mainly spherical, with an average size of (223±51) nm, and zeta-potential of (17.4±6.1) mV. The agarose gel electrophoresis analysis confirmed that they could effectively protect pDNA from degradation against DNase I. Gene transfection in vitro proved that HA/CS/pDNA nanoparticles could be efficiently transfected into chondrocytes of rabbits, the expression of green fluorescent proteins was observed under the fluorescent microscopy, and the transfection efficiency reached as high as (15.450±0.404)%, significantly higher than that of the naked pDNA or the CS/pDNA nanoparticles (P<0.05). Conclusion HA/CS/pDNA nanoparticles were an effective novel non-viral gene transfer vector, which possessed the potential targeting transfection ability on chondrocytes.     

空心加压螺钉治疗股骨颈骨折疗效分析

目的：探讨空心加压螺钉内固定术治疗股骨颈骨折的疗效及术后致股骨头缺血性坏死的相关影响因素。方法：对2003年1月至2009年6月应用空心加压螺钉治疗的96例股骨颈骨折进行回顾性分析,男44例,女52例;年龄21～88岁,平均56.3岁。将患者年龄、性别、骨折类型、骨折复位情况、外伤至手术复位时间与术后骨折不愈合及股骨头缺血性坏死之间的关系进行统计学分析。结果：84例获得随访,时间9～60个月,平均25.4个月。术后出现下肢深静脉血栓2例,骨折不愈合8例,股骨头缺血性坏死11例。术后Harris评分为（86.20±11.00）分,优40例,良32例,可7例,差5例。未移位骨折组和移位骨折组股骨头坏死发生率分别为3.22%和18.87%,两者差异有统计学意义（P=0.037）;解剖复位组和非解剖复位组的股骨头坏死发生率分别为5.00%和20.45%,两者差异有统计学意义（P=0.036）;而不同年龄、性别、手术时间对继发股骨头坏死无明显差异。结论：空心加压螺钉内固定术治疗无移位股骨颈骨折疗效良好,骨折类型及骨折复位情况是影响术后股骨头缺血性坏死的主要因素。对年轻移位的股骨颈骨折患者,应尽可能解剖复位、牢靠内固定,以减少术后股骨头缺血性坏死的发生;对于骨折移位严重的高龄患者,建议行人工关节置换术。     

不采用术中X线定位的后路椎间盘镜髓核摘除术治疗腰椎间盘突出症

目的探讨后路椎间盘镜手术中不采用X线定位的可行性。方法269例腰椎间盘突出症患者行后路椎间盘镜髓核摘除术,不采用X线进行术中定位。根据术前腰椎平片,通过骨性标志的触摸和术中细针穿刺确定手术间隙并建立工作通道,术中可根据手术情况适当调整工作通道位置。结果手术时间30～90min,269例患者中,267例首次置入工作通道定位准确,2例术中发现定位错误（神经根松弛无间盘突出）,移动工作通道完成手术,无需重新切口。术后所有患者腰腿痛症状消失或缓解,复查X线照片未发现手术节段错误。并发马尾神经损伤1例,单纯脑脊液漏4例,切口愈合不良2例。结论参照腰椎平片,通过骨性标志的仔细触摸和术中细针穿刺定位,不采用术中X线定位的后路腰椎间盘镜手术确实可行,并节省术中定位时间,避免患者和医护人员的射线接触。     

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.     

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.     

硫酸氨基葡萄糖和透明质酸钠治疗髌股关节骨关节炎的研究

目的：观察硫酸氨基葡萄糖和透明质酸钠治疗膝关节髌股关节骨关节炎的疗效及安全性。方法：86例髌股关节骨关节炎病人随机分为硫酸氨基葡萄糖组（A组）和透明质酸钠组（B组）,每组43例。A组口服硫酸氨基葡萄糖胶囊每次2粒,每日3次,连续服用12周。B组关节内注射透明质酸钠2mL,1次/周,共5次。随访半年观察其临床疗效及不良反应。结果：硫酸氨基葡萄糖治疗骨关节炎总有效率为83.7%,透明质酸钠组总有效率为86%,两组病人生活质量明显提高,经比较差异无统计学意义（P〉0.05）。A组和B组不良反应发生率分别为2.3%和9.3%,两组差异有统计学意义（P〈0.05）。结论：硫酸氨基葡萄糖和透明质酸钠治疗膝关节髌股骨关节炎均可以改善临床症状,硫酸氨基葡萄糖不良反应较少。     

关节镜下同种异体跟腱骨一期重建膝关节前后交叉韧带

目的 评价关节镜下利用同种异体跟腱骨一期重建膝关节前后交叉韧带的疗效.方法 2000年7月至2005年2月收治15例患膝前后交叉韧带断裂但对侧膝关节完好者,在关节镜下先对合并存在的半月板损伤进行修复,然后使用2条同种异体跟腱骨一期重建前后交叉韧带.亚急性期或慢性期(>3周)重建12例,急性期(<3周)重建3例.手术前后采用IKDC和Lysholm评分系统对患膝关节功能进行评估,随访结果与对侧健康膝火节进行比较.结果 所有患者均获得36～40个月(平均38个月)随访.根据IKDC评分,术前所有患膝关节功能都严重异常,术后9例患膝功能改善为止常,5例接近正常,1例异常.Lysholm评分由术前平均(56±5)分改善为术后(90±4)分,差异有统汁学意义(t=15.660,P<0.05.结论同种异体跟腱骨可用于关节镜下重建膝关节前后交叉韧带,疗效满意.     

卒中单元管理模式联合通心络胶囊治疗急性脑梗死的临床研究

目的探讨卒中单元管理模式联合应用通心络胶囊治疗急性脑梗死的临床疗效。方法选择脑梗死住院患者208例,随机分为对照组（103倒）和联合治疗组（105例）。对照组采用常规内科治疗,联合治疗组在对照组基础上加用卒中单元联合通心络胶囊治疗。治疗前后对两组患者进行神经功能缺损评分;采用简式Fugl-Meyer法评定两组患者治疗前后肢体运动功能;用Barthel指数评定两组患者治疗前后日常生活活动能力（ADL）。结果联合治疗组总有效率高于对照组（P〈0．01）;治疗后,联合治疗组的神经功能缺损评分低于对照组（P〈O．01）;联合治疗组在日常生活活动能力和运动功能改善方面,均优于对照组（P〈O．01）。结论卒中单元联合通心络胶囊能显著改善急性脑梗死患者神经功能,提高患者日常生活活动能力和运动功能,疗效显著,值得临床借鉴。     

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.