5-氮胞苷诱导对骨髓间充质千细胞凋亡影响的实验研究

目的：研究5-胞苷（5-azacytidine，5-aza）对体外培养的兔骨髓问充质于细胞（mensenchymal stemcell，MSC）凋亡的影响。方法：体外分离培养兔MSC，用不同浓度5-aza诱导不同时间．观察MSC凋亡情况。结果：正常培养兔MSC有轻度细胞捌亡；5-aza诱导浓度达15μmol／L时凋亡率明显增加（P〈0．01）．当浓度达10μmol／L，诱导时间延长则细胞洲亡率明显增加（P〈0．05），超过15μmol／l。时出现成片细胞死亡。结论：5-aza诱导对体外培养兔骨髓间质干细胞有诱导细胞凋亡作用，其作用程度与诱导时间及浓度有关。     

骨形态发生蛋白-7对肾、骨的作用

骨形态发生蛋白-7（Bone Morphogenetic Protein-7，BMP-7）属转化生长因子．8（Transforming Growth Factor-β，TGF-β）超家族成员，是一种多效性细胞因子，与肾脏发育、肾功能的维持、各种肾脏疾病密切相关，是肾脏保护因子，重组人骨形态蛋白-7（rhBMP-7）已被认为是肾病治疗的新方法。另外，BMP-7在骨骼发育、骨修复中也具有一定的调节作用，并与多种骨疾病密切相关，尤其在肾性骨营养失调病的肾脏形态功能和骨代谢紊乱性疾病中有密切联系。祖国传统医学“肾主骨”理论认为：肾对骨起调控作用，肾精充足，则骨髓生化有源，骨的生长发育及修复均与肾气滋养有关，BMP-7可能是“肾主骨”理论的物质基础的新依据。本文对BMP-7在肾、骨中的作用做一综述。     

慢性丙型肝炎患者血浆瘦素和肿瘤坏死因子-α水平与肝纤维化的关系

新近报告酒精性肝硬化患者循环瘦素水平增加,瘦素水平还与某些类型肝脂肪变性有关。此外,动物实验显示肝星状细胞也表达瘦素,而星状细胞在肝纤维化发生中又起关键作用。     

血管生成素-2在食管鳞癌中的表达及意义

目的初步探讨血管生成素-2(Angiopoietin-2,Ang-2)在食管鳞状细胞癌组织中的表达及其与肿瘤病理分级、浸润和转移的关系。方法采用S-P免疫组织化学方法,对85例食管鳞癌及22例正常食管组织中的Ang-2表达水平进行检测,并与肿瘤病理分级、浸润深度及淋巴结转移情况进行统计学分析。结果85例食管鳞癌组织中,57例Ang-2表达呈阳性,阳性率为67.06%,显著高于Ang-2在正常食管组织中的表达(P<0.05)。Ang-2表达与肿瘤病理分级和淋巴结转移显著相关(P<0.05)。结论Ang-2的表达与食管鳞癌的临床分期及病理分级呈正相关,提示Ang-2促进肿瘤新生血管形成,参与食管鳞癌的发生和发展,可作为反映食管癌进展的生物学指标。     

部分创面外用抗菌药物与成纤维细胞生长因子2、表皮生长因子、重组人生长激素对成纤维细胞生物学特性影响的实验研究

目的观察部分创面外用抗菌药物与成纤维细胞生长因子(FGF)2、表皮生长因子 (EGF)、重组人生长激素(rhGH)对成纤维细胞生物学特性的影响。方法体外培养成纤维细胞, 按所加药物不同分为对照组(常规培养)、丁胺卡那霉素(0．021、0．210、2．100 mg/L)组、庆大霉素(5、 50、500 mg/L)组、氯霉素(0．01、0．10、1．00 mg/L)组、磺胺米隆(5、10 g/L)组、FGF2(2 400 U/ml)组、 EGF(2 000 U/ml)组及rhGH(0．016、0．160、1．600 g/L)组。用噻唑蓝(MTT)法测定各组成纤维细胞增殖活性[吸光度(A)值],用流式细胞仪检测细胞周期,并于显微镜下观察细胞形态的变化。结果 (1)MTT法检测:与对照组A值0．455 3±0．021 7比较,各种剂量丁胺卡那霉素组、庆大霉素组、氯霉素组、磺胺米隆组成纤维细胞A值均明显降低(P<0．05或0．01),其中磺胺米隆(5、10 g/L)组降低最明显,分别为0．101 3±0．001 1、0．095 0±0．004 1(P<0．01)。FGF2组及0．016 g/L rhGH 组细胞A值明显高于对照组(P<0．05),而EGF组及0.160、1．600 g/L rhGH组A值与对照组接近 (P>0．05)。(2)细胞周期检测:对照组细胞增殖指数(PI)为(9．63±0．45)％,与之比较,0．210 mg/L丁胺卡那霉素组细胞PI值无明显变化(P>0．05),FGF2组、EGF组及0．016 g/L rhGH组PI值均明显升高,分别为(46．76±2．33)％、(42．30±1．41)％、(13．29±0．47)％(P<0．05或0．01)。 (3)形态学观察:对照组、EGF组及0．160、1．600 g/L rhGH组成纤维细胞数目较多,呈长条形或梭形, 轮廓不清,透明度高;丁胺卡那霉素组、庆大霉素组、氯霉素组、磺胺米隆组细胞数目较少,形态不规则,轮廓清晰,透明度低,细胞内多有颗粒样物质及空泡;FGF2组、0．016 g/L rhGH组细胞分布均匀、密集,呈长条形或梭形,核分裂相多见,轮廓不清,透明度高。结论不同创面外用药物对成纤维细胞生物学特性的影响各异,在创面修复过程中应选择合适的创面外用药物,以促进愈合并抑制瘢痕增生。     

45例正常人听觉诱发电位P50的分析

目的 利用诱发电位技术探讨正常人听觉P50特征.方法 应用美国Nicolet Bravo脑电生理仪,采用条件刺激(S1)-测试刺激(S2)模式对45名健康受试者作了听觉P50检测.结果 分析了基本波型,提出了Cz和Pz区域P50诸指标平均值.结论 听觉P50电位具有抑制性特征,其变化可反映大脑正常感觉门的功能状态.P50较佳表达式是波幅S1-S2和100(1-S2/S1)两种结合,能直接显示感觉门的程度.此正常值可供抑郁症研究时参考.     

Effect on peripheral nerve vivo with human insulin-like regeneration by transgene in growth factor-1

BACKGROUND: Human insulin-like growth factor (hIGF-1) has been successful in treating peripheral nerve injury, but it is still unclear whether hIGF-1 after transgene in vivo has the effect on promoting the regeneration of peripheral nerve. OBJECTIVE: To observe the effect of hIGF-1 on the regeneration of peripheral nerve by transgene in vivo with electrophysiology, histological morphology and ultromicro morphology. DESIGN: A univariate design. SETTINGS: Jilin Institute of Surgery, China-Japan Friendship Hospital Affiliated to Jilin University; School of Basic Medical Sciences, Jilin University. MATERIALS: Thirty male adult Wistar rats of grade Ⅱ, weighing 200-250 g, were provided by the Animal Experimental Center of Jilin University [certification number: SCXK-(Ji)20030001]. The rats were raised in the environment at the temperature of 25 ℃ and humidity of 70%. All the rats were randomly divided into hIGF-1-treated group, treatment control group and blank control group, 10 rats in each group. Positive liposomes (mass concentration of 2 g/L) and pcDNA3.1 (mass concentration of 1 g/L) were purchased from Beijing Yuanpinghao Company; pcDNAhIGF-1 (mass concentration of 1 g/L) was provided by Dr. Shen from the School of Public Health of Jilin University. The liposomes were mixed with plasmids with the mass ratio of 1.5 to 10.Operative microscope was made by Jiangsu Zhenjiang Microsurgical Instrument Factory; EMB-5304K electromyogram (EMG) evoked potential meter by Nihon Kohden Corporation. HPIAS-1 000 high-acuity color pathological imaging analytical system (Japan) and JEM-1200EX transmission electron microscope (Japan) were also used. METHODS: The experiments were carried out in Jilin Institute of Surgery from April to June in 2004. ① All the rats were anesthetized, and the right sciatic nerve was exposed, and it was clipped with a clip at 5 mm below the piriform muscle for 3 times, 10 s for each time. The pressed width was 3 mm, and formed as membrane under operating microscope (×6). Rats in the hIGF-1-treated group were subepineurially injected with the mixture of pcDNAhIGF-1 and positive liposomes (10 μL) immediately, those in the treatment control group were injected with the mixture of pcDNA3.1, positive liposomes and distilled water (10 μL), and those in the blank control group were not given any injection. ② The sciatic nerve functional indexes (SFI) were measured within 56 days postoperatively according to the methods used by Shen et al. ISFI=0 was taken as normal, and ISFI=-100 as completely damaged. EMG evoked potential meter was used to record the electrophysiological changes of the regenerated nerve fibers. The indexes of histological morphology in 5 randomly selected sights were determined with the color pathological imaging analytical system, and the ultrostructures of the regenerated nerve fibers were also observed. MAIN OUTCOME MEASURES: ① Comparison of the SFI within 56 days postoperatively; ② Comparison of the electrophysiology, histological morphology and ultrastructure of the regenerated nerve fibers 56 days postoperatively. RESULTS: All the 30 Wistar rats were involved in the analysis of results. ① SFI: The SFI values were gradually increased as time prolonged in all the three groups, and the changes were more obvious after 24 days, the SFI values recovered better at each time point in the hIGF-1-treated group than in the other two groups. ② Eelectrophysiological results of right sciatic nerve: The latency of motor evoked potential (MEP) was close between the treatment control group and the blank control group [(2.55±0.36), (2.65±0.55) ms, P > 0.05], but higher in the hIGF-1-treated group [(2.14±0.22) ms] than in the blank control group (P < 0.01). The amplitude and conduction velocity of MEP in the treatment control group [(6.67±0.69) mV, (29.57±4.06) m/s] were close to those in the blank control group [(6.60±0.59) mV, (29.22±3.20) m/s, P > 0.05], but those in the hIGF-1-treated group [(7.81±0.84) mV, (36.91±4.37) m/s] were larger or faster than those in the blank control group (P < 0.01). ③ Results of the pathological image analysis of the regenerated nerve fibers: The axonal diameter, thickness of myelin sheath of the regenerated nerve fiber and the number of myelinated nerve fiber in the treatment control group [(2.28±0.33) μm, (1.08±0.18) μm2, (71.80±8.25) fibers] were close to those in the blank control group [(2.18±0.29) μm, (1.03±0.15) μm2, (68.60±8.55) fibers] (P > 0.05), and those in the hIGF-1-treated group [(3.03±0.35) μm, (1.65±0.24) μm2, (88.20±8.82) fibers] were obviously larger or more than those in the blank control group (P < 0.01). ④ Ultrastructure of the regenerated nerve fibers of sciatic nerve: In the hIGF-1-treated group, the regenerated fibers of sciatic nerve were more and mature, manifested by thicker nerve fibers, thicker and evener myelin sheath, which were better than those in the other two groups. CONCLUSION: The results of the quantitative parameters of the electrophysiology, gross histological morphology and ultrostructural changes in the process of repairing damaged peripheral nerve indicate that transgene in vivo with hIGF-1 can promote the neural regeneration after peripheral nerve injury.     

缺氧诱导因子-1α在胶质瘤中的研究进展

缺氧诱导因子1(Hypoxiainduciblefactor1,HIF1)是调节细胞缺氧反应的主要转录因子,HIF1α为其活性调节亚单位。HIF1α在胶质瘤中表达显著增加,并与胶质瘤的恶性进展密切相关。HIF1为异二聚体结构,调控多种与细胞缺氧反应相关基因的表达;氧水平、相关癌基因与抑癌基因及细胞因子等均可调节HIF1α蛋白的表达;HIF1α和肿瘤内新生血管形成、细胞代谢、细胞凋亡及细胞坏死之间均存在密切关系,并可影响放化疗对肿瘤的疗效;以HIF1α为靶点的药物或基因治疗正在成为胶质瘤基础和临床研究关注的热点。