Inhibition of transforming growth factor-beta 1 alters the growth, anchor-dependent cell aggregation and integrin mRNA expression in human promonocytes: implications for endometriosis and peritoneal adhesion formation

Transforming growth factor beta (TGF-beta) is a major secretory product of macrophages which, through autocrine/paracrine pathways, play a central role in normal reproductive tissues as well as in disorders such as endometriosis and intraperitoneal adhesion formation. Using TGF- beta antisense oligonucleotides and U937 cells (a promonocytic human cell line) as an in-vitro model, the present study examined the autocrine mediated action of TGF-beta 1 on proliferation, anchor- dependent and -independent cell aggregation and expression of several mRNAs of cell surface adhesion molecules including integrins and platelet-endothelial cell adhesion molecule (PECAM-1). Northern blot analysis and enzyme-linked inmmunosorbent assay (ELISA) revealed that treatment with TGF-beta 1 antisense, but not sense or nonsense oligomers, in a dose-dependent manner (0.1-10 microM) down-regulated the expression of TGF-beta 1 mRNA and protein to undetectable amounts at the highest antisense concentration. TGF-beta 1 antisense at < 1 mM slightly increased, while at > 3 microM significantly inhibited, the rate of DNA synthesis and proliferation of these cells (P < 0.05). Treatment with TGF-beta 1 antisense promoted cell aggregation under anchor-independent culture conditions (plastic dishes), while it suppressed colony formation under anchor-dependent culture conditions (soft agar assay). U937 cells expressed alpha 2, alpha 3, alpha 4, alpha 5, alpha 6, beta 1 and beta 2 integrin mRNA and PECAM-1 mRNA, while alpha v, beta 3 and beta 5 integrin mRNA was undetectable. The relative amount of alpha 2, alpha 3, alpha 4, alpha 6, beta 1 and beta 2 integrin and PECAM-1 mRNA expression were down-regulated in a dose- dependent manner after TGF-beta 1 antisense treatment, while alpha 5 integrin mRNA expression was up-regulated, although it was undetectable at 10 microM antisense. In contrast, TGF-beta 1 antisense up-regulated beta 3 mRNA expression with maximal effect occurring at 10 microM. These results provide evidence that the autocrine loop of monocyte/macrophage-derived TGF-beta 1 action is essential for regulation of growth, aggregation and the expression of adhesion molecules by these cells. We propose that in disorders such as endometriosis and peritoneal fibrous adhesions, significantly higher numbers of tissue macrophages with the capacity to express excess TGF- beta 1 yield an environment able to promote cell-cell and cell-matrix interactions, and thus lead to further complications from these abnormalities. We are currently investigating whether site-specific inhibition of TGF-beta using antisense strategy is a useful tool for management of these lesions, particularly after their post-surgical removal.      

运用蛋白质芯片技术鉴定干扰素的亚型

目的：采用自制的6株干扰素单克隆抗体，在本实验室建立的金基底蛋白质芯片平台上对IFN-α1b、IFN-α2a、IFN-α2b、IFN-β及IFN-γ进行鉴定。方法：金基底蛋白质芯片用已知亚型的IFN半成品包被，封闭后与自制的6株干扰素单抗反应，再与Cy3-羊抗鼠IgG反应，反应后GenePix4100A芯片扫描仪扫描读数，将结果与ELISA法、Western blot法比较。同样方法检测IFN成品。结果：金基底蛋白质芯片检测技术结果表明，该平台鉴定干扰素的亚型具有特异性，与ELISA法、Western blot法能得出一致的结论。结论：运用本实验室建立的蛋白质芯片平台可以对干扰素的亚型进行鉴定，具有高通量，操作简单，样品用量少，重复性强等优点。     

HA-40vol%Ti复合材料与动物骨组织相容性体内研究

采用热压烧结法制备HA-40vol%Ti复合材料,并通过体内植入试验,研究了该复合材料的生物相容性.结果表明,HA-40vol.%Ti复合材料具有良好的生物相容性和骨引导性,可以与骨发生骨整合,早期骨整合和骨引导性优于纯Ti.在整个植入时间段内,HA-40vol.%Ti复合材料与周围骨组织的结合强度均比纯钛的高,且增长的快.植入3月后,复合材料与新骨的结合强度已达到4.73MPa.     

葡萄糖神经酰胺合成酶在人乳腺癌细胞的表达及其与多药耐药的关系

鞘脂类物质神经酰胺通过介导肿瘤细胞凋亡在一定程度上逆转了肿瘤多药耐药(MDR)。人乳腺癌细胞葡萄糖神经酰胺合成酶(glucosylceramide synthase,GCS)使神经酰胺糖基化为葡萄糖神经酰胺(GC),降低了神经酰胺的促凋亡作用,促进了肿瘤细胞MDR。试图经本实验探讨人乳腺癌细胞GCS表达与多药耐药的关系。     

ONO-AE-248诱发中性粒细胞非凋亡性程序化细胞死亡的生化信号转导初探

目的：研究ONO-AE-248诱发中性粒细胞（PMN）非凋亡性程序化细胞死亡过程中，细胞死亡相关的蛋白分子caspase-3、caspase-8、p-38MAPK和PKC的作用，探讨这种细胞死亡的分子机制。方法：运用光镜和DNA电泳对PMN形态学变化和生化特征进行观察和评估；通过Western blot显示PMN中caspase-3、caspase-8活性改变和p-38MAPK磷酸化改变；运用MTS法检测PKC抑制剂对PMN活性的影响。结果：PMN经ONO-AE-248刺激，绝大多数分叶核融合成单叶核（12小时），DNA琼脂糖电泳结果显示缺乏梯状条带（24小时），而且ONO-AE-248对caspase-3、caspase-8活性和p-38MAPK磷酸化水平无明显影响。然而，PKC抑制剂STS和H-7能显著抑制ONO-AE-248对PMN的促死亡效应。结论：ONO-AE-248所诱导的PMN死亡可能是一种非凋亡性的主动性细胞死亡，即非凋亡性程序化细胞死亡。该死亡过程不依赖caspase-3、caspase-8的活化和p-38MAPK的磷酸化，但是PKC可能发挥正向的调控作用。     

基于电生理-力学复合心脏模型的右心室三维运动分析

在原有电生理心脏模型基础上,我们采用有限元方法和复合材料理论建立了一个电生理-力学复合双心室模型,并基于该模型对右心室心壁在收缩期的三维运动进行仿真分析。模型建立过程中,考虑了心肌的纤维旋向、心电兴奋产生的心肌收缩力。心室壁位移、最小主应变(E 3)等用来表征心室的运动。仿真结果与加标记磁共振成像的实验结果基本一致。另外,仿真结果还表明,虽然心基部位移最大,但最大收缩力却发生在心尖部,这个结果用一般的动物实验或人体实验是难以得到的。因此,电生理-力学复合双心室模型对于今后进一步评价心脏功能具有重要意义。     

Predictors of hepatic steatosis in HBeAg-negative chronic hepatitis B patients and their diagnostic values in hepatic fibrosis

Objective: To investigate predictors of hepatic steatosis in HBeAg-negative chronic hepatitis B (CHB) patients and their diagnostic values in hepatic inflammation and fibrosis. Methods: A total of 106 HBeAg-negative CHB patients with clinically and pathologically proven steatosis and 98 patients without steatosis were recruited into this study. The levels of fasting blood glucose (FBG), fasting insulin (FINS), triglyceride (TG), cholesterol (CHOL), alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (Alb), globulin (Glb), HBV DNA, body mass index (BMI), homeostatic model assessment of insulin resistance (HOMA-IR) and pathological changes of the liver in inflammation, fibrosis and fatty deposition were examined in all patients. Results: The levels of BMI, HOMA-IR, FBG, insulin, TG, and CHOL were significantly higher in patients with steatosis than those without steatosis (all P<0.05). But ALT, AST and HBV DNA levels were significantly lower in patients with steatosis (all P<0.05). Logistic regression analysis showed that only FINS was a significant predictor for hepatic steatosis (P<0.05); FINS and Glb were significant predictors for hepatic inflammation (all P<0.05); BMI and TC were significant predictors for hepatic fibrosis (all P<0.05). Conclusions: Hepatic steatosis, a common disease in HBeAg-negative CHB patients, was positively associated with BMI, FBG, FINS, TG, TC, GGT, ALP and HOMA-IR. In these patients, the prevalence of hepatic inflammation and fibrosis was also increased.     

班级生涯心理辅导促进大学生职业生涯规划的对照研究

目的:考察班级生涯心理辅导对大学生职业生涯规划的作用。方法:方便选取某高校2个平行班学生为研究对象,将2个班随机分为实验班(N=32)和对照班(N=28)。对实验班进行为期3个月、共10个单元的"班级生涯心理辅导"课程,对照班不做干预。用班级生涯心理辅导效果评估表,对实验班学生在第5单元(中期评估)和第10单元(末期评估)结束时分别进行测试。辅导结束3个半月后对实验班学生进行追踪访谈。结果:经过3个月的班级生涯心理辅导,实验班学生在职业生涯规划的信心和能力等方面有了显著提高(如职业生涯规划的信心反馈"有很大提高",由中期评估的8.3%提高到末期评估的33.3%)。追踪访谈表明,实验班学生明确了自己的生涯定向,制订出各自的生涯规划方案,有了较清楚的生涯发展目标。结论:班级生涯心理辅导对大学生的职业生涯规划有积极的促进作用。     

小鼠胚胎腭板发育过程中麦胚凝集素、蓖麻凝集素和鸡冠珊瑚树凝集素受体的表达

方法:以生物素标记的凝集素WGA、 RCA和ECL为探针,应用亲和组织化学显色方法检测上述3种凝集素受体在各组小鼠胚胎腭板上皮及其间充质的分布状况和变化规律.结果:在小鼠胚胎腭发育过程中3种凝集素受体在腭板上皮中的表达水平均高于在间充质中的表达水平.WGA受体在腭板上皮中的表达峰值出现于d1322-d148,即腭板上抬和水平生长期;而RCA和ECL受体的表达峰值出现于d158-d1522,即腭板融合期.结论:3种凝集素受体均可能不同程度地参与腭板正常发育的调节,WGA受体可能主要作用于腭板上抬和水平增长期,而RCA和ECL受体则可能主要作用于腭板融合期.     

注意缺陷型和混合型注意缺陷多动障碍儿童的眼跳特征

目的:使用眼跳任务深入研究不同亚型注意缺陷多动障碍(Attention deficit hyperactivity disor-der,ADHD)儿童的抑制损伤。方法:选取金华市2所小学1500人,使用DSM-Ⅳ诊断标准、Conners儿童行为量表(父母问卷和教师问卷),筛查出10～13岁儿童48名,其中注意缺陷型(I型)11人,混合型(C型)19人,对照组18人,全体进行了正向眼跳和反向眼跳。结果:⑴反向眼跳中对照组的错误率低于I型和C型[(37.97±13.14)%vs.(70.90±15.78)%,(84.04±12.84)%;均P0.001],I型的错误率低于C型(P=0.015)。⑵对照组比C型更容易改正错误的眼跳[(95.39±7.49)%vs.(85.54±15.87)%,P0.05],对照组的矫正反应时要短于I型和C型(P0.05)。⑶反向眼跳中C型比对照组和I型精确性更差[(5.47±1.63)°vs.(4.41±1.00)°,(4.23±1.31)°;P0.05],正向眼跳中ADHD儿童有更多的先期性眼跳[(3.11±4.55)%vs.(1.11±1.98)%,P0.05]。结论:研究提示C型和I型的ADHD儿童存在抑制功能损伤,且I型的抑制功能好于C型。