不同胎龄胎儿皮肤和增生性瘢痕中细胞外信号调节激酶5及MAPKK基因表达的变化

目的　探讨细胞外信号调节激酶 5 (extracellular signalregulatedproteinkinase 5 ,ERK5 )及其上游信号分子MAPKK(MEK5 )在不同胎龄的胎儿皮肤和不同形成时期的增生性瘢痕中的基因表达变化规律。方法　用病理学技术检测不同发育时期的皮肤和增生性瘢痕的结构特征后 ,提取 18例不同胎龄 (13～ 32周 )的胎儿皮肤、6例少儿皮肤、16例不同发生时期的增生性瘢痕 (4个月～ 11年 )和 8例正常皮肤组织的总RNA后 ,用RT PCR方法检测ERK5和MEK5在不同组织中的表达变化特征。结果　随着胎龄的增加 ,胎儿皮肤中ERK5和MEK5基因表达水平逐渐降低 ;而在少儿皮肤中 ,基因的转录本含量明显减少 (P <0 0 5 )。在正常皮肤和不同形成时期的增生性瘢痕中 ,MEK5基因表达水平相近 ,而ERK5基因在正常皮肤中的表达水平较低 ,在增殖期和成熟期瘢痕中表达水平明显增强 (P <0 0 1)。结论　在早期妊娠胎儿皮肤中ERK5和MEK5基因的高表达可能是胎儿皮肤创面无瘢痕愈合的机制之一 ,而增生性瘢痕发生和形成也可能与这两种基因表达增强有关。     

p73蛋白在原发性胆囊癌中的表达及意义

目的 研究p73蛋白在原发性胆囊癌中的表达并探讨其与肿瘤生物学行为、预后的关系。方法 应用免疫组化S-P法检测p73蛋白在49例原发性胆囊癌、15例胆囊腺瘤性息肉和12例慢性胆囊炎组织中的表达。结果 p73蛋白在原发性胆囊癌中的阳性表达率为49％(24/49),而在胆囊腺瘤性息肉和慢性胆囊炎中均无表达,具有显著差异(P<0.01)。胆囊癌组织中p73蛋白表达与肿瘤分化程度有关,在低分化癌中有较高表达,与高中分化癌比较,差异有显著性(P<0.05)。与性别、年龄、淋巴结有无转移和Nevin分期无关(P>0.05)。与预后有关,p73蛋白表达阴性病人预后显著优于p73蛋白表达阳性病人,术后1年内死亡者p73阳性表达率为68％(21/31),1～3年死亡组为25％(3/12),而>3年死亡组为0,差异有显著性(P<0.05)。结论 p73蛋白在原发性胆囊癌组织中呈高水平表达,p73基因可能参与了胆囊癌的发生、发展过程,可作为判断原发性胆囊癌分化程度和预后的指标。     

抗结核分枝杆菌培养滤液蛋白10的新型单克隆抗体的制备及其临床应用

目的 制备抗结核分枝杆菌培养滤液蛋白10(CFP-10)特定抗原肽的单克隆抗体,为建立结核病临床早期诊断的免疫学方法奠定物质基础.方法 设计并合成结核分枝杆菌CFP-10第53 ～ 66位氨基酸(14肽AAVVRFQEAANKQK)作为特定的抗原肽序列,经免疫BALB/C小鼠,取其脾细胞和小鼠骨髓瘤细胞SP2/0融合,经酶联免疫吸附试验(ELISA)检测、多次有限稀释亚克隆建立能稳定分泌抗结核分枝杆菌特异抗原肽单克隆抗体的杂交瘤细胞系,纯化该单克隆抗体,进行免疫球蛋白亚型分析,并鉴定分析该单克隆抗体在蛋白质免疫印迹法(Western blotting)、免疫沉淀法和ELISA检测中的应用效果.共检测结核分枝杆菌培养上清样本38份,非结核分枝杆菌培养上清样本20份,结核性胸水样本32份,非结核性胸水样本24份以及健康对照血清样本20份.结果 单克隆抗体免疫球蛋白亚型分析显示该杂交瘤细胞系产生的抗体类型为IgG1和κ型.该抗结核分枝杆菌CFP-10特异性单克隆抗体可成功用于Western blotting和免疫沉淀法分析.ELISA定量检测显示,该特异性抗体检测结核分枝杆菌的敏感性为78.6％ (55/70),特异性为92.2％ (59/64).结论 CFP-10特定抗原肽单克隆抗体用于结核分枝杆菌检测有较高的敏感性和特异性,可用于结核病的早期诊断.     

The prognostic impact of fluctuating levels of C-reactive protein in Brazilian haemodialysis patients: a prospective study.

BACKGROUND: A single elevated C-reactive protein (CRP) value predicts mortality in haemodialysis (HD) patients, but the relative importance of repeated vs occasional positive systemic inflammatory response findings is not known. METHODS: To assess the influence on survival of occasional inflammation, CRP, serum albumin (S-Alb) and fibrinogen were analysed bimonthly in 180 HD patients (54% male, 49+/-14 years). Clinically significant inflammation was defined as CRP >5.1 mg/l, based on the receiver operating characteristics curve for CRP as predictor of death. Based on four consecutive measurements of CRP, patients were assigned into three groups: group 1 (n = 74; 41%), no inflammation (CRP < or = 5.1 mg/l in all measurements); group 2 (n = 65; 36%), occasional inflammation (1-3 measurements of CRP > 5.1 mg/l); and group 3 (n = 41; 23%), persistent inflammation (all measurements of CRP >5.1 mg/l). The nutritional status was evaluated by subjective global assessment (SGA) and body mass index (BMI), and the survival (21 months of follow-up) by Kaplan-Meier curve and Cox model. RESULTS: The median and range of CRP values (mg/l) for group 1, 2 and 3 were: 3.2 (3.2-5.1), 3.6 (3.2-54.9) and 13.8 (5.2-82), respectively (P<0.001), whereas the prevalence of malnutrition, assessed by SGA and BMI, did not differ significantly between the groups. The survival rate by Kaplan-Meier analysis was significantly different among the groups (chi2 = 12.34; P = 0.0004). Patients in group 3 showed the highest mortality (34%; P = 0.001), compared with group 1 (8%) and group 2 (14%; P = 0.01), respectively, whereas there was no significant difference in mortality between groups 1 and 2. Age, CRP, S-Alb level and SGA were independent predictors of mortality. CONCLUSION: The patients with a persistent elevation of CRP had a higher mortality rate than the patients with occasional CRP elevation. Thus, persistent, rather than occasional, inflammation is an important predictor of death in HD patients.     

Ergothioneine pretreatment protects the liver from ischemia-reperfusion injury caused by increasing hepatic heat shock protein 70

BACKGROUND: Reperfusion of the liver after ischemia induces the expression of the heat shock genes and the synthesis of the heat shock proteins (HSP). We studied the effects of the natural antioxidant ergothioneine (EGT) treatment on the expression of HSP70 in ischemic-reperfused (IR) liver. METHODS: Adult male Wistar rats were randomly divided into three groups: Sham group given standard laboratory chow and water for 3 weeks followed by sham operation; Control group given standard laboratory chow and water for 3 weeks followed by liver IR injury; EGT group given standard laboratory chow supplementation l-ergothioneine (1.2 mg/kg/d body weight) administered by gavage and water for 3 weeks followed by liver IR injury. Ten rats from each group were killed to determine serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactic dehydrogenase (LDH), tissue malondialdehyde (MDA), HSP70 levels, and histologic changes at 30, 60, and 120 min of reperfusion, respectively. Survival was followed for 1 week. RESULTS: IR caused significant increase in serum AST, ALT, LDH, and tissue MDA levels. As compared with the control group, animals treated with EGT experienced a significant decrease in serum AST, ALT, and LDH levels in all reperfusion periods. Tissue MDA levels in animals receiving EGT were significantly reduced as compared with control group at 30 min and 60 min after reperfusion. After ischemia, reperfusion caused a remarkable production of HSP70 in the control group. When the rats were pretreated with EGT, the levels of HSP70 increased significantly in their livers after reperfusion compared with the control group. Liver injury in the EGT-treated animals was lower to that in the control group. The 7-day survival rate was significantly improved (from 50% to 80%) by EGT pretreatment. CONCLUSION: HSP70 has been shown to induce tolerance against warm IR injury in rat livers. EGT pretreatment protects the liver from IR injury by over-expression of HSP and the subsequent suppression of lipid peroxidation.     

罗格列酮对多囊肾囊肿衬里上皮细胞p38促分裂原活化蛋白激酶信号通路的影响

Objective To investigate the effect of rosiglitazone on p38 mitogen-activated protein kinase (p38MAPK) pathway in polycystic kidney cyst-lining epithelial cells. Methods The cyst-lining epithelial cells (PKD cells) from human polycystic kidney were treated with rosiglitazone (10 μmol/L), peroxisome proliferator-activated receptor-γ (PPARγ) inhibitor GW9662 (10 μmol/L), rosiglitazone (10 μmol/L) +GW9662 (10 μmol/L), p38MAPK specific inhibitor SB203580 (10 μmol/L), SB203580 (10 μmol/L)+ rosiglitazone(10 μmol/L) for 2 hours followed by epidermal growth factor (EGF) stimulation. Protein expressions of p38, phuspho-p38 (p-p38) and proliferating cell nuclear antigen (PCNA) were detected by Western blot. p38 mRNA was examined by RT-PCR. Expression of c-fos and c-jun was observed by immunocytochemistry. Results (1) EGF markedly up-regulated the expressions of p38, p-p38, PCNA, c-fos anti c-jun compared with control group (P<0.01). (2) Compared with EGF treated group, rosiglitazone significantly reduced p38 activation and mRNA expression (P<0.01, respectively). Rosiglitazone, rosiglitazone+SB203580 could significantly down-regulated p-p38, PCNA, c-fos and c-jun expression (P<0.01, respectively) with no significant difference between these two groups. (3) GW9662 partially reversed the reduction effect of rosiglitazone. Conclusions Rosiglitazone can inhibit proliferation of autosomal dominant polycystic kidney disease cyst-lining epithelial cells partially through down-regulating p38 activation and reducing c-fos, c-jun and PCNA expression. The above effect of rosiglitazone is in part PPARγ-independcnt.     

骨形态发生蛋白复合同种松质骨载体修复节段性骨缺损的实验研究

目的　本实验将探讨同种松质骨作为 BMP的载体修复节段性骨缺损。方法　将 rh BMP- 2复合同种松质骨载体 (其中含 rh BMP- 2 0 .4 mg)植入兔桡骨 15 m m人工缺损处 ,以新鲜自体松质骨植入 ,单纯同种松质骨植入作为对照 ,通过放射学骨缺损修复 L ane评分 ,Nilsson骨愈合组织学评分 ,扫描电镜观察 ,比较术后 4、8、12周各组修复骨缺损的效果。结果　 rh BMP- 2复合同种骨载体组术后 4、8、12周各项评分与同期自体骨组之间无显著性差异 (P>0 .0 5 ) ,明显优于单纯同种骨组 (P<0 .0 5 )。结论　 rh BMP- 2具有高效的骨诱导能力。同种骨是一种较理想的 BMP载体 ,rh BMP- 2复合同种松质骨载体的骨修复效果与自体骨基本一致     

基于胆汁泡相糖蛋白特征而研制诊断胆结石ELISA药盒的可行性分析

目的:分析胆汁泡相糖蛋白的特征并评估其ELISA药盒临床应用的可行性。方法:应用凝集素探针介导的点印迹法分析泡蛋白糖链的组成与结构,利用酶解方法测定泡蛋白各组分的构成,并分析其氨基酸组成。利用Westernblot法分析胆汁与血清泡蛋白的同源性,根据ELISAkit测定泡蛋白的含量分布,并以ROC曲线判别ELISAkit的临床应用价值。结果:化学分析泡蛋白由多肽和寡糖组成,其中多肽分子量约为21u,糖链约占总蛋白含量的37.3%,为核心岩藻糖丰富的多天线复杂型聚糖。泡蛋白具胆汁与血清分布两相性,胆固醇性结石组胆囊胆汁和血清33.5u泡蛋白含量分别为(213.4±70.1)μg/ml、(179.8±97.9)μg/ml,明显高于胆色素性结石症组和正常对照组(P<0.05),而后两者间差异无显著性(P>0.05)。ELISAkit判断胆固醇性结石的胆汁相敏感度和特异度分别为85%和90%,而血清相为61.7%和93.3%。结论:胆汁泡蛋白是一种新认识的胆结石促成核因子,其在不同人群中具有表达谱的异质性,ELISAkit辅助诊断胆石症具有一定的临床应用价值,但尚需更多病例的积累和验证。     

急性坏死性胰腺炎大鼠脑脊液髓磷脂碱性蛋白水平变化的初步研究

目的:研究急性坏死性胰腺炎(ANP)大鼠24h内脑脊液髓磷脂碱性蛋白(MBP)水平的动态变化。方法:应用5%和1%的牛磺胆酸钠分别诱导ANP和急性水肿性胰腺炎(AEP)大鼠模型,同时设立假手术对照组。分别于模型诱发后3、6、9、12及24h,经大鼠小脑延髓池抽取脑脊液。以酶联免疫吸附(ELISA)法测定标本中的MBP含量。结果:诱发ANP后3h,脑脊液MBP水平明显高于AEP组和假手术组(P<0.01);6h和9h时仍维持于较高水平(P<0.05),12h开始缓慢降低。而AEP组和假手术组的MBP水平无明显变化,在24h内维持于较低水平。结论:ANP大鼠早期时脑脊液MBP水平已异常升高,提示脑组织髓鞘结构已有损害。检测脑脊液MBP水平变化对临床胰腺炎病人并发脑功能障碍的诊断和预后评估具有重要意义。     

高频线阵型超声波在肛瘘诊断中的应用

目的：探讨高频线阵型超声波在肛瘘诊断中的价值。方法：使用GE公司LOGIQ7型超声波诊断仪10MHz高频线阵型探头对26例复杂肛瘘及可疑肛瘘患者进行术前检查，并与手术结果对比。结果：超声波显示情况与手术结果完全符合。结论：高频线阵型超声波在肛瘘诊断中具有较高的价值。