半合成抗体库的构建及抗Tie2 Fab抗体的筛选

构建一个半合成抗体库 ,不经免疫制备人源抗Tie2Fab抗体。通过RT PCR方法 ,从人脐带血淋巴细胞总RNA扩增轻链基因及重链VH段基因 ,将轻链基因插入pCOMb3载体中 ,得人轻链质粒库 ;从HBsAb的Fd段基因制备含有不同长度随机化CDR3的FR3 CDR3 J CH1片段 ,然后将VH段基因与随机化的CDR3融合 ,得到Fd基因片段 ,再将其插入轻链质粒库中 ,得半合成人Fab质粒库。通过多次建库 ,获得总容量为 2× 1 0 7的半合成抗体库。其Fd段和轻链基因的重组率为 5 0 %。经 3轮淘洗 ,从噬菌体抗体库中筛选到与Tie2抗原特异结合的噬菌体克隆。

Fas and Bc1-2 EXpression on T Lymphocyte Subsets in the Peripheral Blood of Relapsing Patients with Condyloma Acuminatum

Objective: To study the expression of Fas and Bcl-2proteins on T lymphocyte subsets in the peripheralblood of relapsing patients with condyloma acuminatum(CA) and healthy controls. Methods: Flow cytometry (permeabization andstaining procedure with conjugated antibodies) wasused. Results: We observed that the expression of Fasprotein on CD4^+ T lymphocyte subset of CA patientswas significantly higher than that of healthy controls(P<0.01). Conclusions: Increased expression of Fas proteinon CD4^+ T lymphocyte subset may be a cause of de-creased percentage of CD4^+ T lymphocyte subset. Thisinduces the increased ratio of CD4^+/CD8^+.

LncRNA FAS-AS1转染对K562细胞生物学行为的影响

目的探究LncRNA FAS-AS1对K562白血病细胞生物学行为的影响。方法取对数生长期的K562细胞培养分为空白对照组(正常培养,不做任何处理),阴性对照组(转染LncRNA FAS-AS1空质粒),实验组(Lnc RNA FAS-AS1)。MTT法检测细胞增殖情况,Western blot检测细胞中mTOR、AMPK蛋白含量,流式细胞仪检测细胞凋亡情况。结果与空白对照组和阴性对照组相比,实验组K562细胞FAS-AS1 mRNA表达显著上升(P<0.05),但空白组与阴性对照组细胞中FAS-AS1 mRNA表达比较差异无统计学意义(P>0.05)。随着培养时间的延长,实验组K562细胞的增殖率显著低于对照组(P<0.05),但空白与阴性对照组细胞增殖率比较差异无统计学意义(P>0.05)。实验组凋亡率显著高空白对照和阴性对照组(P<0.05),而空白对照组与阴性对照组凋亡率比较差异无统计学意义(P>0.05)。实验组K562细胞AMPK蛋白表达水平显著高于空白对照组和阴性对照组(P<0.05),而空白对照组和阴性对照组K562细胞中表达水平蛋白mTOR、AMPK比较差异无统计学意义(P>0.05)。结论LncRNA FAS-AS1过表达对K562白血病细胞的生物学行为有显著影响,能够降低细胞的增殖数量及增殖活性,可能与活化AMPK基因进而阻断mTOR活性相关。

基于FXR-Srebp1c-FAS通路及CD36探讨祛痰活血方抗非酒精性脂肪性肝病的机制

目的 以内源性脂肪酸合成相关法尼醇X受体(farnesoid X receptor,FXR)-固醇调节元件结合蛋白-1c(sterol regulatory element-bindingprotein-1c,Srebp1c)-脂肪酸合成酶(fatty acid synthase,FAS)通路及外源性脂肪酸摄入相关的白细胞分化抗原36(cluster of differentiation 36,CD36)为切入点,探讨祛痰活血方抗非酒精性脂肪性肝病(non-alcoholic fatty liver disease,NAFLD)的分子机制。方法 雄性C57BL/6J小鼠,随机分为对照组、模型组、祛痰活血方组和易善复组,每组各10只。模型组、祛痰活血方组和易善复组给予高脂饲料喂养,4周后祛痰活血方组和易善复组开始给药干预,给药8周后取各组小鼠血清用于生化检测;肝组织用于病理、肝脏总胆固醇(cholesterol,TC)、甘油三酯(triglyceride,TG)含量及FXR、Srebp1c、FAS、CD36基因及蛋白表达的检测。结果 与对照组比较,模型组小鼠体质量、肝质量明显增加(P<0.05);肝组织病理结果及肝脏TC、TG含量的升高(P<0.01)均提示明显脂肪变性;肝组织FXR、Srebp1c的m RNA表达水平,以及CD36的m RNA和蛋白表达均明显上升(P<0.01),FAS蛋白表达下降(P<0.05);与模型组比较,祛痰活血组小鼠体质量、肝质量均明显降低(P<0.01);肝组织脂肪变性明显减轻;肝组织中FXR m RNA表达明显升高(P<0.01),Srebp1c、FAS、CD36的m RNA及蛋白表达均明显降低(P<0.05)。结论 外源性脂肪酸摄入增加可能是高脂引起的NAFLD的主要成因,祛痰活血方既抑制外源性脂肪酸摄入,亦减少内源性脂肪酸合成,双向调节改善NAFLD。

基于GA-BP-FA算法的多轴铣削残余应力优化

提出了面向多轴铣削GH4169的残余应力控制的GA-BP-FA多目标优化模型,运用遗传算法(GA)优化BP神经网络的初始权值和阈值,提升神经网络算法的收敛速度和预测精度。基于GA-BP算法建立了铣削残余应力预测模型,并分析了铣削工艺参数对残余应力的影响。以获得最小残余拉应力/最大残余压应力为目标,基于萤火虫算法(FA)对铣削工艺参数进行优化。研究结果表明,所建立的残余应力的GA-BP预测模型具有较高的预测精度,两个方向的平均误差分别为18.9%和16.7%。经FA算法优化后,所选参数阈内最优工艺参数为切削倾角Φ=85°,切削速度v=20 m/min,每齿进给量f_(z)=0.01 mm/z。优化后σ_(x)方向的残余压应力-463.43 MPa和σ_(y)方向的残余压应力-686.93 MPa。

FasL、Caspase-9表达水平对肝细胞性肝癌患者介入治疗后预后的判断价值分析

目的 探讨介入治疗前后肝细胞性肝癌(HCC)患者Fas配体(FasL)、半胱氨酸天冬氨酸蛋白酶-9(Caspase-9)表达变化及意义。方法 选取68例接受介入治疗的HCC患者为研究对象。对比治疗前后HCC患者FasL、Caspase-9水平。统计HCC患者复发情况,并分为复发组和未复发组。对比复发组和未复发组临床资料、治疗前后FasL、Caspase-9水平。Logistic多因素回归分析影响HCC患者治疗后复发的危险因素。制作受试者工作特征曲线(ROC),以曲线下面积(AUC)评价FasL、Caspase-9对HCC患者治疗后复发的预测价值。结果 与治疗前相比,治疗后患者FasL水平降低(P<0.05),Caspase-9水平升高(P<0.05)。随访结束,介入治疗的HCC患者复发发生率为47.06%(32/68)。复发组肿瘤最大直径、肿瘤个数、治疗后FasL水平高于未复发组(P<0.05),治疗后Caspase-9水平低于未复发组(P<0.05)。多因素分析结果显示:肿瘤最大直径(OR=2.619,95%CI:1.126~6.092)、治疗后FasL(OR=2.134,95%CI:1.264~3.602)、治疗后Caspase-9(OR=0.975,95%CI:0.954~0.997)是HCC患者治疗后复发的相关因素(P<0.05)。ROC分析显示,治疗后FasL、Caspase-9及两者联合检测对预测HCC患者治疗后复发灵敏度分别为78.12%(95%CI:59.56%~90.06%)、71.87%(95%CI:53.02%~85.60%)、93.75%(95%CI:77.78%~98.91%),特异度分别为72.22%(95%CI:54.57%~85.21%)、80.56%(95%CI:63.43%~91.20%)、72.22%(95%CI:54.57%~85.21%),AUC分别为0.794(95%CI:0.679~0.883)、0.785(95%CI:0.668~0.875)、0.867(95%CI:0.763~0.937)。结论 介入治疗后HCC患者FasL水平降低、Caspase-9水平升高。治疗后FasL、Caspase-9可作为临床评估HCC患者治疗后复发的重要参考指标。

Expression and significance of Tie-1 and Tie-2 receptors, and angiopoietins-1,2 and 4 in colorectal adenocarcinoma:Immunohistochemical analysis and correlation with clinicopathological factors

AIM: There is strong evidence that tyrosine kinases are involved in the regulation of tumor progression, cellular growth and differentiation. Recently, many kinds of tyrosine kinase receptors have been reported, among them Tie-1 and Tie-2 receptors constitute a major class.Angiopoietin (Ang)-1 is known as a ligand ofTie-2 tyrosine kinase receptor. The objective of this study was to establish a comprehensive Tie-1 and Tie-2 and Ang-1, 2 and 4expression profile in human colorectal adenocarcinomas.METHODS: We examined 96 cases of surgically resected human colorectal adenocarcinoma by immunohistochemistry and investigated the statistical correlation between the expressions of Ties and Angs and clinicopathological factors.RESULTS: Among the 96 cases of adenocarcinoma, 87(90.6%), 92 (95.8%), 83 (86.5%), 89 (92.7%), and 76cases (79.2%) showed positive staining in the cytoplasm of carcinoma cells for the Tie-1 and Tie-2 and Ang-1, 2and 4 proteins, respectively. Histologically, the expressions of Ties and Angs were variable. The expressions of Ties and Angs were correlated with several clinicopathological factors, but did not correlate with the presence of lymph node metastasis. Ties and Angs were highly expressed in human colorectal adenocarcinoma cells.CONCLUSION: These findings suggest that the Tie-Ang receptor-ligand complex is one of the factors involved in the cellular differentiation and progression of human colorectal adenocarcinoma.

脓毒症诊治中Ang/Tie信号通路的研究进展

脓毒症以及伴随的多器官功能障碍是一组常见的临床综合征,其患病率随着人口老龄化逐年上升。全球每年发生2000万例脓毒症,死亡率为26%。早期准确诊断脓毒症,有利于及时采取针对性的治疗策略,这是降低病死率的关键。内皮细胞是外源病原体或内源性损伤信号的早期作用对象。研究提示,内皮细胞的结构改变与功能活化在脓毒症的发生、发展中发挥重要作用。作为内皮细胞专属信号通路,血管生成素(angiopoietin,Ang)/酪氨酸激酶受体(tyrosine kinase receptor,Tie)在内皮细胞的异常激活和损伤中发挥重要作用。Ang/Tie信号通路主要包括2种位于内皮细胞中的酪氨酸激酶受体(Tie1、Tie2)和4种分泌性糖蛋白配体(Ang-1、Ang-2、Ang-3、Ang-4)。机制研究方面,Ang-1可持续性激活Tie2受体,维持细胞与细胞、细胞与基质间的相互作用,支持血管内皮细胞正常的生理功能;Ang-2是Ang-1拮抗剂,可竞争性阻断Ang-1与Tie2受体结合。脓毒症环境下,Ang-1下降,Ang-2升高,Ang-1/Ang-2的比值下降。Ang-2竞争性阻断Ang-1与可溶性Tie2受体结合,内皮细胞处于严重异常激活状态。Ang-2介导肝素酶的释放导致糖萼损伤,可引起血管通透性增加;Ang-2可促进炎症反应;Ang/Tie信号系统失调介导凝血功能障碍,Ang-2升高是弥散性血管内凝血的前哨性事件。临床病情监测方面,Ang-2>5.61 ng/mL,诊断脓毒症的灵敏度为74.36%;Ang-2持续升高,提示内皮功能难以恢复,器官发生功能性改变。早期动态监测Ang-2可用于预测脓毒症相关肺损伤、急性肾损伤。Ang-2/Ang-1比值上升、Ang1/可溶性Tie2比值下降可预测脓毒症患者90 d病死率(ROC曲线面积分别为0.787和0.704)。Ang-2和可溶性Tie-2水平下降,则提示脓毒症血浆置换有效。靶向Ang/Tie信号通路的动物实验获得一定成功,但目前临床试验未能获得有价值的结果。脓毒症相关Ang/Tie信号通路有待于进一步深入研究。

Correlation between the Expression of Fas, Bcl-2 in Peripheral Blood Lymphocytes and the LeveL of IFN-γ,IL-4 in Serum of Patients with Condyloma Acuminata

In order to investigate the correlation between the expression of the apoptotic regulatory proteins (Fas, Bcl 2) in peripheral blood lymphocytes (PBLC) and the level of IFN γ and IL 4 in serum of the patients with condyloma acuminata (CA) in the immune pathogenesis of CA, indirect immunofluorescence labeling method of flow cytometer and solid sandwich ELISA method were performed for detecting the expression of Fas, Bcl 2 in PBLC and the level of IFN γ and IL 4 in serum of 60 cases of CA. The results showed the expression level of Fas in PBLC of CA was significantly higher than in the normal control group, but the expression level of Bcl 2 was significantly lower (both P< 0.01). The level of IFN γ in serum of CA was significantly lower than in the normal control group ( P< 0.01), but IL 4 was significantly lower (both P< 0.01). The expression of Fas in PBLC had a negative correlation with the level of IFN γ in serum of patients with CA, but had a positive correlation with the level of IL 4; The expression of Bcl 2 had a positive correlation with the level of IFN γ, but had a negative correlation with the level of IL 4. All the correlation coefficients had significant differerce by t test ( P< 0.01). It was suggested abnormal apoptosis in PBLC, the suppressed secretion of the THl associated cytokines (eg: IFN γ) and the increased secretion of the TH2 asssociated cytokines (eg: IL 4) existed in the patients with CA and might play an important role in the immune pathogenesis of CA.

基于Fas/FasL介导的凋亡信号通路研究通管方对输卵管炎性不孕模型大鼠的作用机制

目的:探讨通管方对输卵管炎性不孕(SI)模型大鼠输卵管组织中Fas/FasL通路的影响。方法:采用经阴道内接种混合菌液的方法,建立输卵管炎性不孕大鼠模型。造模后,77只SD雌性大鼠随机分成空白组、假手术组、模型组、妇可靖胶囊组(0.29 g·kg^(-1)·d^(-1))、通管方低剂量组(7.515 g·kg^(-1)·d^(-1))、通管方中剂量组(15.03 g·kg^(-1)·d^(-1))、通管方高剂量组(30.06 g·kg^(-1)·d^(-1)),各组大鼠分别灌胃给药28 d后处死并取材。采用蛋白免疫印迹法(WB)检测SI模型大鼠输卵管组织中Fas、FasL蛋白及Caspase-1的表达水平,酶联免疫吸附法(ELISA)检测SI模型大鼠血清IL-4、IL-10、Caspase-1的表达变化。结果:与空白组比较,模型组血清中IL-4和IL-10的水平均明显降低,血清和输卵管组织中的Caspase-1水平明显升高(P<0.01);与模型组比较,通管方各剂量组和妇可靖胶囊组降低了血清和输卵管组织中的Caspase-1的表达(P<0.01),上调了血清中IL-4和IL-10水平、输卵管组织中Fas和FasL蛋白的表达(P<0.05,P<0.01)。其中,通管方高剂量组的作用均明显优于其他剂量组和妇可靖胶囊组(P<0.05,P<0.01)。结论:通管方可能通过调控Fas/FasL凋亡信号通路而消除SI模型大鼠输卵管炎症。

Expression of angiopoietins, Tie2 and vascular endothelial growth factor in angiogenesis and progression of hepatocellular carcinoma

AIM： TO investigate the significance of angiopoietins, Tie2 and vascular endothelial growth factor （VEGF） expression in the angiogenesis and progress of hepatocellular carcinoma （HCC）. METHODS： Fresh surgically resected specimens of HCC and noncancerous liver （NCL） tissue from 38 patients with HCC were obtained, and expression of angiopoietin-1 （Ang-1）, angiopoietin-2 （Ang-2）, Tie2, and VEGF messenger RNA （mRNA） was examined by real-time quantitative reverse transcription-polymerase chain reaction （RT-PCR）. Expression pattern of each gene in HCC and NCL tissue specimens was compared and the potential role and interaction in angiogenesis of HCC were analyzed. Genes＇ expression level and its relationship with tumor＇s clinicopathological parameters were also investigated. Immunohistochemical staining of CD34 was performed to determine the microvessel density （MVD） and Ang-2/Ang-1 ratio was calculated. Relationships between Ang-2/Ang-1 ratio, VEGF and MVD and clinicopathological features were also tested so as to evaluate their significance in the progression of HCC. RESULTS： Ang-2 and VEGF mRNAs in HCC were significantly higher than those in NCL tissue （P 〈 0.05）, whereas the Ang-1 and Tie2 mRNAs showed no statistical significance （P 〉 0.05）, though slightly lower level of Ang-1 mRNA in HCC was observed. Ang-2/ Ang-1 ratio and VEGF were both positively correlated to MVD. The Ang-2/Ang-1 ratio, Ang-2 and VEGF were all associated with tumor＇s clinicopathological parameters （P 〈 0.05） except for histological grades （P 〉 0.05）. Ang-1 and Tie2 levels in different clinicopathological groups were not significantly different （P 〉 0.05）. CONCLUSION： Dominant Ang-2 expression against Ang-1 through Tie2 receptor in the presence of VEGF plays a critical role in initiating early neovascularization and transformation of noncancerous liver to hepatocellular carcinoma. Its consequently constant operation in formed HCC induces further angiogenesis and progression of HCC.

基于ikPCA-FABAS-KELM的短期风电功率预测

为了增强在短期风电功率预测领域中传统数据驱动机器学习模型的精度,提出基于ikPCA-FABAS-KELM的短期风电功率预测模型.首先,对主成分分析进行改进,提出可逆核主成分分析(ikPCA),在保证数据特征的同时,降低输入数据的复杂度,以提升模型运行速度;其次,引入萤火虫个体吸引策略对天牛须算法(BAS)进行改进,提出FABAS算法;最后,利用FABAS算法对核极限学习机(KELM)的正则化参数C和核参数γ进行寻优,降低人为因素对模型盲目训练的影响,提高模型预测精度.仿真结果显示,提出的预测模型有效提高了传统模型的预测精度.

Effect of IFNα-2a on Fas expression and apoptosis rate of peripheral blood cytotoxic T cells in patients with hepatitis B

Interferon(IFN) with antiviral and im-munomodulatory activities is one of the most important therapeutic agents for the treatment of chronic hepatitis. The apoptotic effect of IFN is influenced by cell type and the types of IFN, which suppresses proliferation and induces apoptosis in some cell types while inhibiting apoptosis in others. The aim of this study was to explore the effect of IFNα-2a on Fas expression and the apoptosis rate of peripheral blood cytotoxic T cells (CTLs) in patients with hepatitis B. METHODS:Peripheral blood mononuclear cells were isolated from 26 patients with hepatitis B including 16 patients with chronic hepatitis B and 10 patients with chronic severe hepatitis B. Fas expression and apoptosis rate of CTLs were analyzed with flow cytometry before and after IFNα-2a treatment. RESULTS:Before IFNα-2a treatment, Fas expression and apoptosis rate of CTLs from patients with chronic hepatitis B were significantly higher than those from patients with chronic severe hepatitis B and healthy controls respectively. No significant difference was observed between Fas expression and apoptosis rate of CTLs from patients with chronic severe hepatitis B and healthy controls. After IFNα-2a treatment,Fas expression and apoptosis rate of CTLs from different groups were compared with those before IFNα-2a treatment, showing no significant difference despite alternation of different degree. CONCLUSIONS:Activation induced cell death (AICD) exists in peripheral blood CTLs from patients with hepatitis B. No effect of IFNα-2a exerts on Fas expression and apoptosis rate of Fas in patients with hepatitis B.

益母草注射液通过其生物碱成分上调Tie-2、VEGFR2和VEGF促进斑马鱼血管新生

目的:研究益母草注射液在斑马鱼血管损伤模型中的促血管生成作用,并阐明潜在的物质基础和分子机制。方法:采用HPLC-PAD和TLC技术检测益母草注射液的主要成分;利用舒尼替尼诱导转基因Tg(flk1:eGFP)斑马鱼血管损伤模型,益母草注射液干预治疗后,测量斑马鱼节间血管(ISVs)长度,QT-PCR检测血管生成相关基因的表达水平。结果:发现益母草注射液的主要成分是水苏碱、胆碱和葫芦巴碱;其中,益母草注射液、葫芦巴碱和胆碱可减轻化学诱导的斑马鱼血管损伤,其作用机制涉及益母草注射液上调Tie-2、VEGF和VEGFR2的表达,其主要成分葫芦巴碱上调Tie-2和VEGF的表达,胆碱上调VEGF和VEGFR2的表达。结论:益母草注射液及其主要生物碱成分可通过上调Tie-2、VEGFR2和VEGF来促进血管生成。

CHANGES OF SOLUBLE FAS AND SOLUBLE FAS LIGANDIN SERUM AND PERITONEAL FLUID OF INFERTILEPATIENTS WITH ENDOMETRIOSIS

Objective To evaluate the relationship between levels of soluble Fas(sFas)and soluble Fas ligand(sFasL)in serum and peritoneal fluid of endometriosis-associated infertility. Methods The soluble Fas ligand and soluble Fas levels in serum and peritoneal fluid of 20 infertile patients with endometriosis were assessed with enzyme-linked immunosorbent assay, and were compared with 14 infertile patients due to chronic pelvic infectious disease and 16 fertile controls. Results The sFasL levels were significantly higher in infertile patients with endometriosis(175.09 ± 80.55 pg/mL in serum and 284.50 ± 152.38 pg/mL in peritoneal fluid)than those of infertile controls (88.47 ± 43.55 pg/mL in serum and 17.30 ± 9.62 pg/mL in peritoneal fluid)and fertile controls(16.13 ± 11.75 pg/mL in serum and 8.84 ± 2.31 pg/mL in peritoneal fluid). In contrast, as for the sFas levels, infertile patients with endometriosis(828.60 ± 429.65 pg/mL in serum and 349.61 ± 288.89 pg/mL in peritoneal fluid)did not show any significant difference compared with those in infertile patients resulting from pelvic infectious disease(868.75 ± 570.48 pg/mL in serum and 181.76 ± 157.78 pg/mL in peritoneal fluid)and fertile control(822.26 ± 129.12 pg/mL in serum and 318.42 ± 145.16 pg/mL in peritoneal fluid). Conclusions Based upon these results, high level of sFasL in serum and peritoneal fluid and thus apoptosis mediated by it may be implicated in the mechanism involved in endometriosis-related infertility.

鱼类肠道益生地衣芽孢杆菌FA6发酵工艺的响应面法优化

研究以地衣芽孢杆菌FA6活菌数为指标,采用单因素实验和响应面法实验优化了地衣芽孢杆菌FA6发酵培养基和发酵条件,获得了地衣芽孢杆菌FA6最佳发酵培养基:70 g/L可溶性淀粉、40 g/L酪蛋白胨、2 g/L K2HPO4、1 g/L KH_(2)PO_(4)、2 g/L Na_(2)HPO_(4)、2 g/L MgSO_(4)和8 g/L NaCl;以及最佳发酵条件:初始pH 7.0、接种量1%、发酵温度28.8℃、发酵时间31h、装液量50 mL/250 mL和转速160 r/min。在此发酵培养基和发酵条件下,地衣芽孢杆菌FA6活菌数为7.97×10^(9)CFU/mL,较优化前提高了8.1倍。最佳发酵工艺的获得,将能够为鱼源芽孢杆菌的高密度生产,产业化应用提供基础。

Serum levels of soluble Fas, nitric oxide and cytokines in acute decompensated cirrhotic patients

AIM： To evaluate plasma levels of nitrite/nitrate （NOx）, soluble Pas （spas） antigen, tumor necrosis factor alpha （TNF-α） and interleukin-6 （IL-6） in patients with compensated and acute decompensated cirrhosis and to evaluate mediators causing acute decompensation in liver cirrhosis, METHODS： This prospective study was conducted in the medical intensive care unit of an academic tertiary center, Fifty-five patients with acute decompensation （gastrointestinal hemorrhage, encephalopathy, hydropic decompensation） and twenty-five patients with compensated liver drrhosis were included, Blood samples were taken for analyses of spas, Nox, IL-6, TNF-α, Liver enzymes and kidney functions were also tested, RESULTS： In patients with acute decompensation, plasma spas levels were higher than in non-decompensated patients （15305 ± 4646 vs 12458± 4322 pg/mL, P 〈 0.05）. This was also true for the subgroup of patients with alcoholic liver cirrhosis （P 〈 0.05）. The other mediators were not different and none of the parameters predicted survival, except for ALT （alanine-aminotransferase）. In patients with portal-hypertension-induced acute hemorrhage, NOx levels were significantly lower than in patients with other forms of decompensation （70.8 ± 48.3 vs 112.9 ± 74.9 pg/mL, P 〈 0.05）. When NOx levels were normalized to creatinine levels, the difference disappeared. IL-6, TNF-α and spas were not different between bleeders and non-bleeders. In decompensated patients spas, IL-6 and NOx levels correlated positively with creatinine levels, while IL-6 levels were dependent on Child class. CONCLUSION： In acute decompensated cirrhotic patients sPas is increased, suggesting a role of apoptosis in this process and patients with acute bleeding have lower NOx levels, However, in this acute complex clinical situation, kidney function seems to have a predominant influence on mediator levels,

Gene Expression of Fas,Soluble Fas and Fas-Ligand in Thyroid Tissues and Thyrocytes from Patients with Graves′ Disease

ObjectiveTo investigate Fas,soluble Fas(sFas)and Fas ligand(Fas L)gene expression in thyroid tissues and thyrocytes from patients with Graves disease(GD)and to find the interrelationship between apoptosis and pathogenesis of GD. MethodsThyroid tissues were obtained from 7 GD patients and 3 healthy subjects who died accidentally. Thyrocytes were cultured in Eagle′s medium. Total RNA was isolated from thyroid tissues and cultured thyrocytes. The cDNA was prepared by reverse transcription and amplified for Fas,sFas and Fas L by polymerase chain reaction(PCR). ResultsFas and sFas mRNA were detected in all samples from both GD and normal thyroid tissues and thyrocytes,but Fas L mRNA was only found in GD thyroid tissues and thyrocytes. Semi quantitative analysis showed that when compared with those of normal controls,the Fas and sFas mRNA levels were markedly increased in GD thyroid tissues(P<0.01),whereas in GD thyrocytes only the sFas mRNA levels was significantly elevated(P<0.01). ConclusionGene expression of Fas,sFas and Fas L showed abnormality in both thyroid tissues and thyrocytes from GD. The increased production of sFas might be involved in the hyperplasia of thyroid gland.

Association of Fas/Apo1 gene promoter (-670 A/G) polymorphism in Tunisian patients with IBD

AIM： To detect a possible association between the polymorphism of the （-670 A/G） Fas/Apol gene promoter and susceptibility to Crohn＇s disease （CD） and ulcerative colitis （UC） in the Tunisian population. METHODS： The （-670 A/G） Fas polymorphism was analyzed in 105 patients with CD, 59 patients with UC, and 100 controls using the polymerase chain reaction restriction fragment length polymorphism method. RESULTS： Significantly lower frequencies of the Fas -670 A allele and A/A homozygous individuals were observed in CD and UC patients when compared with controls. Analysis of （-670 A/G） Fas polymorphism with respect to sex in CD and UC showed a significant difference in A/A genotypes between female patients and controls （P corrected = 0.004 ＂in CD patients＂ and P corrected = 0.02 ＂in UC patients＂, respectively）. Analysis also showed a statistically significant association between genotype AA of the （-670 A/G） polymorphism and the ileum localization of the lesions （P corrected = 0.048） and between genotype GG and the colon localization （P corrected = 0.009）. The analysis of IBD patients according to clinical behavior revealed no difference. CONCLUSION： Fas-670 polymorphism was associated with the development of CD and UC in the Tunisian population.

基于Bow-Tie-LOPA的煤矿安全风险评估方法应用研究

为解决传统安全评估方法的局限性,有效降低煤矿安全事故、提升煤矿安全管理效能,提出基于Bow-Tie模型的LOPA煤矿安全风险评估方法,通过研究两种方法的概念并分析两种方法相结合的应用,综合应用Bow-Tie-LOPA方法进行煤矿安全风险评估,分析验证Bow-Tie-LOPA方法在煤矿安全风险评估中的可行性和有效性;利用Bow-Tie模型绘制煤矿安全风险事件因果链,找出可供LOPA方法分析的事故场景,并基于LOPA分析风险事件发生率和后果,利用风险矩阵和确定风险等级的方法,进一步评估事故场景,以确定煤矿安全风险后果以及安全保护措施。研究结果表明:LOPA方法的引入有利于Bow-Tie模型分析煤矿复杂事故和危机情境,提高煤矿安全风险管理水平。