反义寡核苷酸对血管平滑肌细胞c—myc,增殖细胞核抗原 …

目的 研究反义ｃ－ｍｙｃ、增殖细胞核抗原（ＰＣＮＡ）基因片断对血管平滑肌细胞（ＶＳＭＣＳ）增殖的作用。方法 使用正义、反义及错配反义ｃ－ｍｙｃ、ＰＣＮＡ,分别艇于体外培养的ＶＳＭＣＳ,蛋白印迹和免疫组化法测定燕经计算机图像分析,观察其对ｃ－ｍｙｃ、ＰＣＮＡ蛋白表达的影响。结果 １０μｍｏｌ／Ｌ浓度的反义ｃ－ｍｙｃ、ＰＣＮＡ作用于ＶＳＭＣＳ后１￣５ｄ相庆ｃ－ｍｙｃ和ＰＣＡＮ蛋白表达减弱,计算机图像分     

肺癌中耐药基因p170的表达及其临床意义

目的：检测耐药基因ｐ１７０在要前有、无化疗的肺癌中表达情况以探讨其临床意义。方法；采用Ｓ－Ｐ免疫组化方法。检测１１３例肺癌（其中术前化疗的肺癌９３例，术前曾化疗的肺癌２０例）和１０例肺组织中ｐ１７０的表达水平。结果：在１１３例肺癌中ｐ１７０阳性表达率为４６％，其中小细胞癌（ＳＣＬＣ）（１５．４％）明显低于非小细胞癌（ＮＳＣＬＣ）（５０％）。术前曾化疗的肺癌２０例，ｐ１７０阳性表达率为７５％。术前未     

鼠转化生长因子-β1基因的克隆及其在成骨细胞中的表达

目的 克隆鼠转化生长因子－β１（ＴＧＦ－β１）基因并研究其在转染的成骨细胞中表达情况。方法 采用逆转录－聚合酶链反应（ＲＴ－ＰＣＲ）可从大鼠淋巴细胞中扩增出１．２ｋｂ特异性片段,经酶切鉴定证实为鼠ＴＧＦ－β１ ｃＤＮＡ。将此片段插入５．４Ｋｂ的ｐｃＤＮＡ表达载体,构建得到６．６Ｋｂ的ｐｃＤＮＡ－ＴＧＦ－β１重组质粒。应用脂质体介导的基因转移技术将ＴＧＦ－β１表达载体导入鼠成骨细胞,４８ｈ后用ＳＡＢ     

乳癌中p16,c-erbB-2的表达及意义

探讨ｐ１６和ｃ－ｅｒｂＢ－２蛋白表达与乳癌临床病理因素及预后的关系。方法 采用免疫组化和ＰＣＲ－ＳＳＣＰ技术,分别检测５０例乳癌ｐ１６,ｃ－ｅｒｂＢ－２蛋白的表达和２０例乳癌ｐ１６基因的纯合性缺失及突变。结果 ５０例乳癌中,１７例（３４．０％）ｐ１６蛋白表达阳性,２４例（４８．０％）ｃ－ｅｒｂＢ－２蛋白表达阳性。２０例乳癌中无ｐ１６基因的纯合性缺失,１例ｐ１６基因外显子２有点突变。结果表明,ｐ１６     

PSP94-TNFα D11a融合基因直接注射的抗肿瘤作用

目的：探讨人ＰＳＰ９４和肿瘤坏死因子α衍生物１１ａ（ＴＮＦαＤ１１ａ）融合基因（ＰＳＰ９４－ＴＮＦαＤ１１ａ）直接注射的抗肿瘤作用。方法：人前列腺癌裸鼠模型肌肉注射含ＰＳＰ９４－ＴＮＦαＤ１１ａ融合基因的ｐｃＤＮＡ－ＰＳＰ９４－ＴＮＦαＤ１１ａ质粒ＤＮＡ,剂量为５０μｇ／只,同时设ｐｃＤＮＡ－ＰＳＰ９４、ｐｃＤＮＡ３．０空载体、生理盐水和环磷酰胺对照组。注射后第２０ｄ处死动物,称瘤重计算抑瘤率。结     

p21 ras和nm23-H1表达与胃癌淋巴结转移的关系

为探讨ｐ２１ｒａｓ和ｎｍ２３－Ｈ１表达与胃癌淋巴结转移的关系,采用ＬＳＡＢ法观察了８０例胃癌患者的ｐ２１ｒａｓ和ｎｍ２３Ｈ１的表达,并分析其与淋巴结转移的关系,结果：有淋巴结转移（ＬＮＭ）组ｐ２１ｒａｓ表达阳性率（６２．５％）明显高于无ＬＮＭ组（４２．５％）但ｎｍ２３－Ｈ１表达阳性率（２７．５％）则明显低于无ＬＮＭ组（４７．５％）ＬＮＭ数为１－３枚和ｐ２１ｒａｓ表达阳性率（３３．３％）明显低于４－     

慢性胆囊炎和肿瘤的关系

目的探讨慢性胆囊炎后壁肥厚所致的腺上皮增生、异型、癌变的发生、发展过程。方法对１０５例慢性胆囊炎壁肥厚和１３例慢性胆囊炎壁肥厚型腺癌,应用ＳＰ免疫组织化学方法,观察ｐ５３、ｐ１６、Ｂｃｌ－２基因表达。结果慢性胆囊炎引起囊壁结节状增厚,腺上皮增生、异型,最后导致壁肥厚型腺癌。ｐ５３在慢性胆囊炎壁肥厚轻、中度中不表达,慢性胆囊炎壁重度肥厚和胆囊炎后壁肥厚型腺癌中阳性表达７．４％和４６．２％;ｐ１６在慢性胆囊炎壁肥厚轻、中、重度和胆囊炎后壁肥厚型腺癌中阳性表达分别为８２．６％,７５．０％,５１．９％和３８．５％,Ｂｃｌ－２在慢性胆囊炎壁肥厚轻、中、重度和胆囊炎后壁肥厚型腺癌中阳性表达分别为３９．１％,４６．９％,７４．１％和４６．２％。结论ｐ５３与ｐ１６、Ｂｃｌ－２基因表达差异有显著意义（Ｐ＜０．０１）;ｐ１６和Ｂｃｌ－２基因表达有显著的相关性;重度的慢性胆囊炎壁肥厚是重要的癌前病变。     

体外免疫吸附对狼疮肾炎的治疗和保护作用

目的　观察体外免疫吸附（ＥＩＡ） 对狼疮肾炎（ＬＮ） 的治疗和肾脏保护作用。方法　比较树脂型ＥＩＡ、大剂量激素和ＣＴＸ联合治疗（ 简称ＥＩＡ组）与大剂量激素和ＣＴＸ治疗（ 简称对照组） 血ＩｇＧ、ＡＮＡ、ｄｓＤＮＡ、Ｃ３、２４ 小时尿蛋白、ＢＵＮ、Ｓｃｒ 及其他临床指标的变化。结果　治疗２ 周时,ＥＩＡ组血ＩｇＧ、ＡＮＡ、ｄｓＤＮＡ显著低于对照组［ 两组分别为ＩｇＧ（１０－０３ ±２－９１） 、（１３－５４ ±３－０５）ｇ／Ｌ,ＡＮＡ（１４－００ ±１３－１４）、（３１－８２ ±３０－０２）ｔｉｔｅｒ－ １,ｄｓＤＮＡ（８－３０ ±４－８３） 、（２７－００ ±１８－５１） ％］ 。Ｃ３ 显著回升［ 两组分别为（５８７－３±９７－０３） 、（４８１－５±１３２－０４）ｍｇ／Ｌ］,２４ 小时尿蛋白、ＢＵＮ、Ｓｃｒ 显著低于对照组［ 两组分别为２４ 小时尿蛋白（２－２８±２－０７） 、（５－９８ ±３－３４）ｇ／２４ｈ,ＢＵＮ（９－１７ ±２－８３）、（１５－２９ ±５－３８）ｍｍｏｌ／Ｌ、Ｓｃｒ（１５６－７５ ±８９－３３）、（２４７－１４±７４－６９）μｍｏｌ／Ｌ） 。治疗４ 周时,ＥＩＡ 组血ＩｇＧ、ｄｓＤＮＡ 仍低于对照组［ 两组分别为Ｉｇ     

HESW与抗癌药物对膀胱癌细胞C－myc癌基因表达的影响

利用国产液电碎石机（ＥＳＷＬ－Ⅲ／２Ｍ型）产生的高能冲击波和１０－羟基喜树碱单纯或联合处理人膀胱癌细胞系（ＢＴ５６３７）。实验分对照组，ＨＥＳＷ组，药物组和联合组。冲击波条件：２００ＳＷ（１０ｋＶ，６０次／ｍｉｎ），药物浓度（０．１μｇ／ｍｌ），水温３７℃。采用α－３２Ｐ－ｄＣＴＰ标记Ｃ－ｍｙｃ癌基因探针，通过Ｎｏｒｔｈｅｒｎ印迹杂交技术，分析四组癌细胞Ｃ－ｍｙｃ癌基因表达。结果表明：对照组Ｃ－ｍｙｃ癌基因表达明显，其转录产物为２．７ｋｂ，而实验各组Ｃ－ｍｙｃ癌基因表达完全被抑制。结论：ＨＥＳＷ，１０－羟基喜树碱及其联合对人膀胱癌细胞的生长抑制作用与Ｃ－ｍｙｃ癌基因表达有密切关系。     

逆转录病毒介导人白细胞介素2基因在人膀胱癌细胞中高效表达

构建人白细胞介素２（ｈＩＬ－２）基因逆转录病毒表达载体ｐＺｉｐ－ｈＩＬ－２，用ＰＡ３１７包装细胞系包装后获得高效价逆转录病毒（１～５×１０５ｃｆｕ／ｍｌ）。病毒感染人膀胱癌细胞系ＢＩＵ８７并用Ｇ４１８选择培养，得到Ｇ４１８抗性克隆；经小鼠脾母细胞增殖法测定、ＲＮＡＳｌｏｔ打点杂交实验和Ｗｅｓｔｅｒｎ免疫印迹实验证实其高效表达ｈＩＬ－２，平均活性在２４小时每毫升为７５Ｕ／１０６细胞（最高达１２５Ｕ／ｍｌ）。转ｈＩＬ－２基因在人膀胱癌细胞中的高效表达，可为人膀胱癌转基因瘤苗的应用研究提供实验基础。     

感觉性和运动性神经来源许旺细胞神经生长因子的表达

目的;研究感觉性和运动性许旺细胞神经生长因子的表达是否有区别,进而探讨对神经特异性再生的影响。方法：体外培养高纯度的运动性许旺细胞,每种细胞分别在常规和添加白细胞介素－1培养基中培养,用双抗体夹心ELISA法测量不同时间培养培养基中神经生长因子表达量。结果：两种培养条件一,感觉性许旺细胞的神经生长均呈双峰状,第二峰较第一峰明显增高,运动性许旺细胞的表达在第6、7d达峰值,但总体水平较感觉性许旺细胞为低。结论：两种许旺细胞神经生长因子的表达量和特点具有差别,这种差别可以为解释神经特异性再生提供依据。     

腺病毒介导的LacZ基因靶向性导入脊髓及示踪周围神经的动态过程

目的由损伤的周围神经靶向性导入腺病毒介导的LacZ基因(AdLacZ)至脊髓，动态观察病毒裁体逆行输送到脊髓前角运动神经元、脊髓后根神经节(dorsal root ganglia，DRGs)感觉神经元及基因产物顺行标记周围神经的全过程和特点。方法分别将AdLacZ转染大鼠正中神经和胫神经近断端，然后以10-0无创线吻合神经。在转染后9周内的24个不同时间点取出正中神经组的C5～T1脊髓节段、DRGs连同臂丛，胫神经组的L2～L6脊髓节段、DRGs连同骶丛。将脊髓和DRGs的50μm横切片行X—gal染色和免疫组织化学染色，臂丛和骶丛的整个标本分别行X—gal染色。计数阳性脊髓前角运动神经元、DRGs神经元及周围神经轴突数，研究转基因表达在脊髓前角运动神经元、DRGs神经元及正中神经、胫神经的最早时间、高峰时间和持续时间。结果LacZ基因能特异性、高效表达在损伤周围神经的感觉和运动神经元。转基因在脊髓和周围神经的表达严格限于感染神经的同侧。正中神经组标本各部位的转基因表达均早于胫神经组。胫神经组被标记的运动神经元和感觉神经元数均高于正中神经组。表达持续的时间在运动神经元最短，然后是感觉神经元，在周围神经持续时间最长。在同一组内，转基因表达在DRGs神经元最早．然后是运动神经元，最后是周围神经干，而且被标记的感觉神经元数多于运动神经元数。结论由损伤的周围神经导入的AdLacZ不但在靶神经元高效特异性表达，而且能高效顺行标记神经元突起、周围神经直至吻合口远端的再生轴突。这对周围神经损伤的基因治疗和神经示踪研究有实用价值。     

双侧下颌升支矢状截骨下齿槽神经损伤和感觉恢复

目的：应用主观和客观测试神经感觉功能恢复的方法，检测双侧下颌升支矢状截骨（bilateralsagittal aplit osteotomy，BSSO）后下齿槽神经（inferior alveolar nerve，IAV）暂时和永久性神经损伤的发生率和感觉恢复的时间。方法：选择14例双侧下颌升支矢状截骨的IAN，在术前和术后7d，1，3，6和12个月采用针刺检测、两点分辨、直流感应电测法决定IAN的损伤和感觉的恢复。结果：IAN暂时性损伤率为78%(22/28)，单侧IAN永久性感觉丧失占7％（2/28）。结论：IAN经损伤术后需6-12个月感觉完全恢复，术中IAN断离可致永久性神经损伤。     

Selectivity of distal reinnervation of regenerating mixed motor and sensory nerve fibres across muscle grafts in rats

This study investigated target specificity during axonal regeneration of a mixed motor and sensory nerve towards respective targets. The femoral nerves in rats were divided and allowed to grow across a 6 mm gap interposed with frozen and thawed muscle grafts towards their distal motor and sensory nerve stumps. Fourteen weeks later the number of motoneurons projecting axons into the motor and sensory branches were determined by retrograde axonal tracing using horse-radish peroxidase. There were significantly higher numbers of motoneurons (p = 0.0034) projecting into the motor nerve than the sensory nerve. Efferent axons of a mixed nerve selectivity grew into motor branches when allowed to regenerate across a 6 mm gap interposed with muscle grafts. It is possible that a deliberately created 'structured gap' during repair of mixed nerves could improve axonal matching by allowing expression of neurotropism.     

失神经支配红白肌肉早期运动神经营养活性研究

目的了解失神经支配后红白肌肉内脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)早期含量变化及相应运动神经传导功能恢复情况,探讨靶器官对损伤运动神经再生修复的作用. 方法 Wistar大鼠40只,平均分为5组,其中1组为对照,其余4组剪断双下肢坐骨神经制作肌肉失神经支配模型,分别于神经损伤后1、3、7和14天采用免疫组织化学方法检测肌肉内BDNF含量.另取15只Wistar大鼠,钳夹双下肢坐骨神经制作失神经支配肌肉神经再生模型,于伤后7天和14天采用神经电生理方法测定神经传导速度及肌电图. 结果比目鱼肌(soleus,SOL)失神经支配后1、3和7天BDNF含量较对照组大幅度升高,差异有统计学意义(P＜0.01),但随时间推移呈下降趋势,14天BDNF含量与对照组接近(P＞0.05);腓肠肌(gastrocnemius,GAS)失神经支配后1、3、7和14天BDNF含量与对照组均接近(P＞0.05).失神经支配肌肉神经再生模型中,伤后7天失神经支配SOL和GAS均未测到动作电位;伤后14天支配GAS和SOL的运动神经传导速度分别恢复至正常值的(36.60±7.40)%和(42.50±3.50)%,二者差异无统计学意义(P＞0.05);GAS M波波幅恢复至正常值的(19.9±6.4)%,而SOL M波波幅恢复至正常值的(13.7±4.0)%,二者差异无统计学意义(P＞0.05). 结论失神经支配红、白肌肉内早期运动神经营养因子含量在不同时间内发生不同的变化,但支配两种靶器官的损伤运动神经功能恢复相近.失神经支配后靶器官的运动神经营养活性是其内部各种运动神经营养因子综合作用的结果.     

The influence of motor axon misdirection on muscle contraction force in early nerve repair in a rat sciatic nerve model.

We examined the influence of misdirection of regenerating motor axons toward the distal sensory Schwann tubes on the muscle contraction force in early nerve repair using a rat sciatic nerve model. At 0, 1, 2, 4 and 8 weeks after severing the tibial, peroneal and sural nerves, the proximal stump of the tibial nerve was anastomosed with the distal stumps of both the peroneal and sural nerves using tubulisation (n=10 in each of five groups). We intentionally used the distal stump of the sural nerve (a sensory nerve) to induce regeneration in motor axons from the proximal tibial nerve stump toward the distal sensory nerve stump. Twenty-four weeks after nerve repair, isometric contraction force and wet weight of the anterior tibial muscle were measured, and the numbers of regenerated myelinated axons (motor and sensory) in the distal sural and peroneal nerves were counted. The rates of sural nerve regeneration were significantly higher at weeks 0 and 1 than at the later repair times. However, muscle contraction force and muscle wet weight did not differ significantly between groups in which nerves were repaired within four weeks of severance. These results indicate that peripheral nerve repair within four weeks of severance does not influence the muscle contraction force of single muscle despite the misdirection of motor axons toward the distal sensory Schwann tubes.     

陈旧性神经损伤后雪旺细胞的变化实验研究

目的 观察大鼠坐骨神经损伤后雪旺细胞的变化。方法 切断成年SD大鼠右侧坐骨神经，形成10mm缺损。于术后1～12个月不同时间段取材。标本用p75受体（p75 neurotrophin receptor．p75NTR）、S-100免疫荧光染色，激光共聚焦显微镜观察，另部分标本进行超微结构观察。结果 神经损伤后1个月，损伤远端神经中p75NTR和S-100的表达最多，在损伤后6个月时p75NTR降至正常水平，S-100则在损伤后9个月时消失。电镜下，雪旺细胞出现在神经内膜管内，神经内膜管下及内膜管周围有大量胶原原纤维增生。结论 大鼠坐骨神经损伤后，雪旺细胞中p75NTR的表达增加，但随损伤时间的延长呈进行性下降，伤后6个月时消失。     

Percutaneous radiofrequency lesioning of sensory branches of the obturator and femoral nerves for the treatment of hip joint pain

BACKGROUND AND OBJECTIVES: The sensory innervation of the hip joint includes the sensory articular branches of the obturator and femoral nerves. In this report, we retrospectively evaluated 14 cases in which hip joint pain was treated by percutaneous radiofrequency lesioning of sensory branches of obturator and/or femoral nerves. METHODS: Fourteen patients who had hip joint pain and underwent percutaneous radiofrequency lesioning of sensory branches of obturator and/or femoral nerves were studied. In all cases, intra-articular hip joint block or articular branch block of obturator nerve with local anesthesia was transiently effective. Radiofrequency lesioning was performed at 75 degrees C to 80 degrees C for 90 seconds using an RFG-3B generator and Sluijter-Mehta cannulae kit (Radionics, Burlington, MA) for the obturator nerve in 9 patients and for both the obturator and femoral nerves in 5 patients. To assess pain intensity, a visual analog scale (VAS) was used. RESULTS: The VAS scores before and after the radiofrequency lesioning were 6.8 +/- 0.9 and 2.7 +/- 1.3, respectively. Twelve patients (86%) reported at least 50% relief of pain for 1 to 11 months. There were no side effects or motor weakness observed. CONCLUSIONS: Percutaneous radiofrequency lesioning of sensory branches of the obturator and femoral nerves is an alternative treatment in patients with hip joint pain, especially in those where operation is not applicable.     

理化法快速鉴别周围神经束性质

目的 探索一种快速鉴别感觉与运动神经束的方法. 方法 纯种Wistar大鼠30只,处死后取整条脊柱.取脊神经节及背根纤维、腹根纤维及坐骨神经,采用免疫印迹实验观察Annexin V 和 Agrin特异性.其中15只大鼠作为实验组,取脊神经前根、后根、坐骨神经及其肌支、皮支行免疫组织化学染色;余15只用PBS代替一抗作为对照组.12月龄健康新西兰大白兔16只,采用喇曼光谱法对不同性质的神经束进行研究. 结果 Annexin V 和 Agrin 分别是感觉神经与运动神经中的特异物质.运动神经与感觉神经的喇曼光谱在 1088、1276、1439、1579 及 1659 cm-1等附近的喇曼振动峰相对强度存在明显差异,1276 cm-1 处的峰值与 1439 cm-1 处的峰值比Ⅰ1 276/Ⅰ1 439 分别为 0.95±0.06 和 1.17±0.08,提示运动神经与感觉神经间的微观结构差异(P<0.05). 结论 显微激光喇曼光谱法是一种鉴别神经束性质的有效的物理学方法,优于免疫组织化学法,具有临床应用的可行性.     

Increased risk of median nerve dysfunction in floor cleaners: a controlled clinical and neurophysiological study.

We studied median nerve involvement in a group of asymptomatic handworkers at risk for carpal tunnel syndrome, and we evaluated damage to thin and thick nerve fibres in the distribution area of the median nerve. Considering floor cleaners as workers at high risk of developing cumulative traumatic disorders in the wrist, we included 42 cleaners and 41 controls. We assessed nerve conduction studies, vibration threshold, and temperature and pain thresholds of the median nerve. The cleaners had significantly impaired motor nerve conduction velocity (p = 0.006), longer sensory distal latency (p = 0.01), lower sensory amplitude (p = 0.0005), and increased difference in heat and cold threshold of the median nerve (p = 0.0002). Increased temperature threshold was associated with prolonged sensory distal latency of the median nerve in the cleaners. In conclusion, impaired neurophysiological variables in the median nerve in floor cleaners compared with controls confirm the hypothesis that those workers are at risk of developing median nerve dysfunction. Sensory nerves seem to be more susceptible to injury than motor branches.