抗病毒治疗慢性丙型肝炎患者T细胞表面程序性死亡1和程序性死亡配体1的表达

目的 观察慢性丙型肝炎(CHC)患者抗病毒治疗24周时外周血CD4+和CD8+T淋巴细胞(T细胞)表面表面程序性死亡1 (PD-1)和程序性死亡配体1(PD-L1)表达水平,分析其与抗病毒治疗临床转归的关系.方法 24例CHC患者,均采用聚乙二醇干扰素α-2a (Peg-IFN α-2a)每周皮下注射一次,联合利巴韦林800 ～ 1200 mg/d,治疗24 ～ 48周.采用流式细胞术和实时荧光定量检测患者治疗前、治疗4、12、24周外周血CD4+和CD8+T细胞表面PD-1、PD-L1表达水平和外周血HCV RNA,全自动生化分析仪检测ALT.采用SPSS16.0软件.两样本计量结果分析采用t检验,治疗前后的计量结果采用重复测量的单因素或两因素方差分析,所有检验为双侧检验. 结果 CHC患者治疗后4周HCV RNA阴性者19例,CD4+和CD8+T细胞表面PD-1的表达率在治疗前分别为18.6％±6.1％和16.6％±13.8％,治疗24周时分别为10.3％±7.7％和9.4％±4.6％,治疗前后比较,PD-1的表达明显下降,F值为12.406和4.955,P值为0.002和0.039,差异有统计学意义.CD8+T细胞表面PD-L1的表达率在治疗前为17.5％±13.7％,治疗4、12、24周时分别为25.9％±11.1％、29.6％±15.1％、32.0％±15.7％,治疗后明显升高,F值分别为9.063、8.365、9.736,P值均＜0.01.治疗4周时,HCV RNA阳性者5例,仅发现CD8+T细胞表面PD-L1的表达治疗24周(39.2％±15.6％)与治疗前(17.4％±16.7％)比较明显升高,F=10.292,P=0.033.持续病毒学应答者20例:CD4+T细胞表面PD-1的表达在治疗4、12、24周分别为14.4％±7.5％、14.0％±6.9％、10.7％±7.6％,治疗前为20.2％±7.5％,与治疗前比较明显下降,F值分别为6.133、5.541、14.780,P＜0.05或P＜0.01.CD8+T细胞表面PD-1的表达在治疗12、24周分别为10.2％±4.6％和10.1％±4.9％,治疗前为16.8％±13.4％,治疗前后比较,PD-1的表达在治疗后明显下降,F值为4.964和4.613,P值均＜0.05.CD8+T细胞表面PD-L1的表达在治疗12、24周分别为30.8％±16.6％和35.2％±16.5％,治疗前为19.0％±14.5％,治疗后明显升高,F=6.442,P=0.020和F=12.349,P=0.002.复发组4例,各治疗时间点PD-1和PD-L1与治疗前比较,差异无统计学意义.结论 快速有效的抗病毒治疗可以下调CHC患者外周血CD4+和CD8+T细胞表面PD-1的表达,上调CD8+T细胞表面PD-L1的表达.CHC患者外周血CD4+和CD8+T细胞表面PD-1和PD-L1表达水平的变化可能与患者抗病毒治疗临床转归存在关系.     

芪灵合剂对慢性乙型肝炎患者T细胞表面程序性死亡1和程序性死亡配体1表达的影响

目的：观察中药芪灵合剂对慢性乙型肝炎（CHB）患者外周血CD4＋T淋巴细胞表面程序性死亡1（Programmed death-1,PD-1）及程序性死亡配体1（Programmed death-1 ligand1,PD-L1）表达水平的影响。方法：收集56例CHB患者随机分为芪灵合剂联合拉米夫定片（LAM）治疗组29例和单用LAM对照组27例,于抗病毒治疗的第0、24、48周用流式细胞术检测外周血CD4＋T淋巴细胞及其表面的PD-1/PD-L1的表达情况。结果：治疗48周后,治疗组患者HBV DNA转阴率、ALT复常率均高于对照组,与对照组比较,HBV DNA转阴率具有统计学意义（P〈0.05）。治疗组患者外周血CD4＋T细胞表面PD-1 24周、48周表达水平与治疗前比较逐渐下降;48周时与本组治疗前比较差异有统计学意义（P〈0.01）,与对照组比较差异有统计学意义（P〈0.05）。治疗组患者CD4＋T细胞PD-L1 48周后表达水平与治疗前比较差异有统计学意义（P〈0.01）,与对照组比较差异有统计学意义（P〈0.05）。结论：中药芪灵合剂联合LAM能够提高CHB患者HBV DNA转阴率,ALT复常率,其作用机制可能与抑制CD4＋T淋巴细胞表面PD-1/PD-L1的表达有关。     

活动性类风湿关节炎调节性T细胞程序性细胞死亡蛋白-1/程序性死亡配体1表达及与临床指标的相关性

目的探讨程序性细胞死亡蛋白-1/程序性死亡配体1(PD-1/PD-L1)在活动性类风湿关节炎(rheumatoid arthritis,RA)患者外周血调节性T细胞表面表达变化及临床意义。方法用流式细胞术分别检测62例活动期RA患者外周血CD4~+CD25~+T细胞和CD4~+CD25~-T细胞群PD-1和PD-L1的阳性率,并设立39例健康体检者作为健康对照组,分析PD-1/PD-L1与实验室各指标和病情活动性的相关性。结果 PD-1在24例高度活动组和38例中低度活动组RA患者CD4~+CD25~+T细胞的表达水平分别为[(2.27±0.56)%和(1.83±1.08)%],高于健康对照组(1.42±0.49)%,差异有统计学意义(P0.01)。PD-L1在高度活动组和中低度活动组RA患者CD4~+CD25~+T细胞的表达水平分别为[(2.06±0.62)%和(1.83±0.63)%],亦高于健康对照组(0.80±0.43)%,与对照组相比差异有统计学意义(P0.001)。PD-1在CD4~+CD25~+T细胞表达水平与DAS28呈正相关(r=0.477,P0.01),PD-L1在CD4~+CD25~-T细胞表达水平与DAS28无相关性(r=0.078,P=0.549)。结论 PD-1/PD-L1在活动性RA患者调节性T细胞表达存在差异,其与DAS28相关性提示PD-1/PD-L1信号通路可经调节性T细胞调控参与RA的发生。     

冠心病患者外周血调节性T细胞及程序性死亡受体-1表达的意义

目的:通过观察冠心病患者调节性T细胞(Tregs)和树突状细胞(DCs)数量和程序性死亡受体-1(PD-1)及其配体(PD-L1)的表达变化,探讨其功能状态,为动脉粥样硬化及不稳定斑块形成的病因及免疫疗法提供思路。方法:入选冠心病患者39例,其中稳定型心绞痛(SA)23例、急性冠状动脉综合征(ACS)16例;20例非冠心病患者为对照组。采用流式细胞术测定各组外周血CD4+CD25hi CD127low PD-1+Treg数量和CD11c+PD-L1+DC数量;用ELISA法测定血清IL-2水平。结果:外周血Treg数量在ACS组、SA组和对照组间差异无统计学意义;ACS组Treg PD-1表达率和DC、PD-L1表达率与SA组和对照组相比明显降低,差异有统计学意义(P<0.05)。结论:PD-1/PD-L1表达下调提示Treg和DC免疫抑制功能缺陷,可能是促进动脉粥样硬化病变进展及不稳定斑块形成的因素。     

外周血T细胞表面PD-1在慢性乙型肝炎患者干扰素治疗期间的表达

目的:观察慢性HBV感染(CHB)免疫清除期PD-1的表达状态以及干扰素抗病毒治疗期间T细胞PD-1的表达,明确免疫清除期CHB患者PD-1表达的临床意义.方法:伴有长期ALT反复升高的CHB患者20例被纳入本研究,所有病例均接受为期6 mo的Peg干扰素(1.5μg/kg)抗病毒治疗,每周1次.应用流式细胞术对所有患者抗病毒治疗不同时间点(0、12、24 wk)的外周血总CD8+T细胞和CD4+T细胞PD-1表达百分比和表达强度进行检测.结果:免疫清除期CHB患者外周血总CD8+T细胞和CD4+T细胞PD-1阳性率和PD-1表达荧光强度均明显高于健康对照组(P<0.05);PEG干扰素治疗早期CD8+T细胞PD-1阳性率呈现明显下降,且CD8+T细胞PD-1表达阳性率同ALT水平和病毒水平均呈现明显正相关(R2=0.22,0.357,P=0.001,0.002).结论:CHB患者外周血T细胞PD-1呈高表达:干扰素治疗诱导的病毒水平下降可导致CD8+T细胞PD-1表达明显下降.     

HBeAg导致慢性乙型肝炎患者外周血Th1/Th2型细胞因子失衡

目的 探讨HBeAg对慢性乙型肝炎(CHB)患者外周血单个核细胞(PBMC)功能的调节作用. 方法 以重组的HBeAg体外刺激CHB患者和健康志愿者的PBMC,用流式细胞术和酶联免疫吸附试验法检测其刺激前后Th1/Th2型细胞因子的变化情况,并观察HBeAg对CHB患者PBMC表面细胞程序性死亡受体(PD)1及其配体(PD-L)1表达的影响.两组间资料比较采用独立样本t检验; PD-1/PD-L1表达水平与HBV DNA拷贝数的相关性采用Spearman相关分析.结果 HBeAg刺激后可使HBeAg阴性CHB患者和健康志愿者CD3+CD4+T淋巴细胞内干扰素(IFN)γ表达水平(0.17％±0.08％与0.17％±0.04％)明显低于未刺激组(0.30％±0.16％与0.32％±0.12％),t值分别为-2.382和-4.190,P值均＜0.01;培养上清液中白细胞介素(IL)-6、IL-10和肿瘤坏死因子α含量明显高于未刺激组(HBeAg阴性CHB患者的t值分别为2.504,3.583和4.324,健康志愿者t值分别为3.542,6.246和5.273,P值均＜0.01).HBeAg刺激PBMC后,HBeAg阴性CHB患者和健康志愿者CD14+细胞表面PD-L1表达水平分别为13.02％±4.98％和3.10％±2.47％,明显高于未刺激组的5.89％±1.56％和0.97％±0.83％,t值分别为4.815和3.454,P值均＜0.05.基础状态下在HBeAg阳性CHB患者外周血中,CD3+CD4+T淋巴细胞内IFNγ表达水平为0.23％±0.09％,明显低于HBeAg阴性CHB患者和健康志愿者的0.34％±0.15％和0.35％±0.09％(t=-3.177,P＜0.01 ; t=-4.541,P＜0.01);而IL-4表达水平为0.39％±0.16％,明显高于HBeAg阴性CHB患者和健康志愿者的0.26％±0.12％和0.23％±0.12％,t值分别为3.382和4.393,P值均＜0.01.基础状态下在HBeAg阳性CHB患者外周血中,CDB+T淋巴细胞表面PD-1和PD-L1表达水平明显高于HBeAg阴性CHB患者及健康志愿者(P值均＜ 0.01),CD14+T淋巴细胞表面PD-L1表达水平显著高于HBeAg阴性患者和健康志愿者,t值分别为5.092和5.473,P值均＜0.01 ; HBeAg阴性CHB患者外周血中CD3+T淋巴细胞表面PD-L1表达水平明显高于健康志愿者(t=3.214,P＜0.01).结论 HBeAg可以明显抑制Th1型细胞因子IFN γ的产生,促进Th2型细胞因子IL-6和IL-10分泌,上调外周血PBMC表面PD-1/PD-L1的表达,从而有利于形成对HBV感染的免疫耐受.因此,HBeAg可能是造成慢性HBV感染者体内免疫耐受的重要因素之一.     

不同HBeAg状态下慢性乙型肝炎患者肝组织内CD4~+、CD8~+、CD20~+、CD57~+ T淋巴细胞的表达分析

目的探讨HBe Ag阳性和阴性慢性乙型肝炎(CHB)患者肝内免疫活性细胞(CD4+、CD8+、CD20+、CD57+T淋巴细胞)与肝脏病理及临床的关系。方法选取2008年1月-2013年12月在桂林市第三人民医院住院的70例CHB患者进行肝穿刺活组织检查,其中HBe Ag阳性患者38例,HBe Ag阴性患者32例。运用免疫组化染色分析肝组织内CD4+、CD8+、CD20+、CD57+T淋巴细胞的表达,比较不同HBe Ag状态患者CD4+、CD8+、CD20+、CD57+T淋巴细胞与临床相关性。等级资料采用Kruskal-Walls H检验,组间比较采用Nemenyi法,相关性分析采用直线相关分析法。结果 HBe Ag阳性和阴性患者的炎症程度到达G3~G4,其肝组织中CD8+、CD20+T淋巴细胞的表达均比G1组明显增加(P值均0.05);HBe Ag阳性患者炎症程度到达G4、而HBe Ag阴性患者炎症程度到达G3,其肝组织中CD4+T淋巴细胞的表达比G1组明显增加(P值均0.05);在HBe Ag阴性患者中CD4+、CD8+、CD20+、CD57+T淋巴细胞均与ALT水平呈正相关(r值分别为0.353、0.628、0.693、0.540,P值均0.05),且CD20+T淋巴细胞还与HBV DNA水平呈正相关(r=0.378,P0.05)。而在HBe Ag阳性患者中只有CD8+、CD20+T淋巴细胞与ALT水平呈正相关,与HBV DNA水平无相关性。结论不同HBe Ag状态患者肝组织内CD4+、CD8+、CD20+T淋巴细胞表达与肝组织内炎症分级均有明显相关性,但肝内免疫活性细胞与ALT、HBV DNA水平的相关性存在差异。     

二至和肝方对肝郁肾虚型慢性乙型肝炎患者PD-1/PD-L1表达的影响

目的:观察二至和肝方联合恩替卡韦治疗肝郁肾虚型慢性乙型肝炎患者外周血中CD4⁺ T细胞表面程序性死亡受体1(PD-1)/程序性死亡配体1(PD-L1)表达水平的影响。方法:将59例慢性乙型肝炎患者随机分为两组,观察组患者29例,予以二至和肝方+恩替卡韦片治疗;对照组患者30例,单用恩替卡韦片治疗。两组患者分别于治疗前、治疗24周、治疗48周后检查外周血丙氨酸氨基转移酶(ALT)、乙型肝炎病毒(HBV)标志物、HBV DNA和CD4⁺ T细胞表面PD-1/PD-L1表达水平。结果:治疗24、48周时两组患者外周血CD4⁺ T细胞表面PD-1/PD-L1表达水平均较本组基线值显著下调(P<0.01)。两组患者治疗前、治疗后24周CD4⁺ T细胞表面PD-1/PD-L1表达水平无明显差异(P>0.05);治疗48周后观察组患者CD4⁺ T细胞表面PD-1/PD-L1表达水平较对照组下降明显(P<0.05)。在治疗24、48周时,观察组患者HBV DNA转阴率显著高于...     

恩替卡韦治疗后慢性乙型肝炎患者外周血T淋巴细胞表面程序性死亡受体1表达与HBeAg血清学转换的关系

目的 动态观察慢性乙型肝炎患者恩替卡韦抗病毒治疗后不同时期外周血T淋巴细胞(简称T细胞)表面程序性死亡受体1(PD-1)表达的变化,并探讨其与HBeAg血清学转换间的关系.方法 对20例HBeAg阳性慢性乙型肝炎患者予以恩替卡韦抗病毒治疗并随访51周,根据HBeAg是否发生血清学转换分为:HBeAg未转换组(14例),HBeAg转换组(6例).分别于治疗前(基线,T0)、治疗2～4周(T1)、治疗5～10周(T2)、治疗11～20周(T3)、治疗21～30周(T4)、治疗31～51周(T5)收集外周血,流式细胞术检测CD4+、CD8+T细胞表面PD-1的表达水平,实时荧光定量PCR检测血清HBV DNA载量,同时检测血清ALT水平.正态分布资料采用独立样本t检验,非正态分布者采用Mann-Whitney U检验比较组间差异,相关性分析采用Pearson相关分析.结果 治疗前两组患者血清HBV DNA载量分别为(7.54±0.67)log10拷贝/ml、(7.30±0.79)log10拷贝/ml(P＞0.05),ALT水平为(187.26±184.15)U/L、(272.17±215.07)U/L(P＞0.05),外周血CD4+T细胞表面PD-1表达水平为6.04%±3.71%6.77%±2.88%(P＞0.05),CD8+T细胞表面PD-1表达水平为6.39%±3.33%、8.88%±2.84%(P＞0.05).恩替卡韦抗病毒治疗后两组患者血清HBV DNA载量、ALT水平的下降与CD4+、CD8+T细胞表面PD-1表达的下调呈显著正相关(r=0.212,P=0.05;r=0.377,P＜0.01;r=0.279,P＜0.05;r=0.347,P＜0.01).在相同的随访时间段内,HBeAg转换组血清HBV DNA载量、ALT水平及外周血CD4+、CD8+T细胞表面PD-1表达的下降率均高于HBeAg未转换组,且两组间△ T0～T1、△T0～T2期HBV DNA的下降率及△T0～T2、△T0～T3期CD8+T细胞表面PD-1表达的下降率差异有统计学意义(分别为49.9%对比37.3%,56.7%对比47.4%,70.1%对比-4.2%,66.9%对比24.5%,P值均＜0.05).结论 HBeAg阳性慢性乙型肝炎患者经恩替卡韦抗病毒治疗后,外周血CD8+T细胞表面PD-1表达的快速下调与血清HBV DNA相似,可作为预测后期HBeAg血清学转换的指标之一.

Abstract：

Objective To observe longitudinally the expression of Programmed death 1 (PD-1) on peripheral blood T cells in chronic hepatitis B patients underwent antiviral treatment with entecavir (ETV)and to explore the relationship between PD-1 expression and HBeAg seroconversion.Methods Twenty HBeAg positive patients underwent antiviral treatment with ETV were followed up for 51 weels.14 patients remained HBeAg positive and 6 patients achieved HBeAg seroconversion.Peripheral blood was collected at six time points:T0:baseline,T1:2-4week;T2:5-10week;T3:11-20week;T4:21-30week:T5:31-51week.PD-1 expressions on T cells were assessed by flow cytometry.Serum HBV DNA loads were determined by real-time fluorescent quanttative polymerase chain reaction (PCR) and serum ALT levels were examined at the same time.Results At baseline,serum HBV DNA load of patients without HBeAg seroconversion and with HBeAg seroconversion were (7.54 ± 0.67) log10 copies/ml and (7.30 ± 0.79) log10 copies/ml(P ＞ 0.05),the ALT levels were (187.26 ± 184.15) U/L and (272.17 ± 215.07) U/L (P ＞ 0.05),PD-1 exprissions on CD4+ T cells were 6.04% ± 3.71% and 6.77% ± 2.88% (P ＞ 0.05),PD-1 exprissions on CD8+ T cells were 6.39% ± 3.33% and 8.88% ± 2.84% (P ＞ 0.05).After ETV treatment,serum HBV DNA loads and ALT levels both decreased gradually,which was positively correlated with PD-1 expressions on CD4+ and CD8+ T cells (r = 0.212,P = 0.05;r = 0.377,P ＜ 0.01;r = 0.279,P ＜ 0.05;r = 0.347,P ＜ 0.01 ).During the same monitoring period,the HBV DNA loads,ALT levels and PD-1 expressions on T cells of the patients with HBeAg seroconversion decreased significantly as compared with the patients without HBeAg seroconversion.Besides,the decrease of HBV DNA loads during period △ T0-T1 and △ T0-T2 and PD-1 expressions on CD8+ T cells during period △ T0-T2 and △ T0-T3 were significantly different between these two kinds of patients (49.9% vs 37.3%,P ＜ 0.05;56.7% vs 47.4%,P ＜ 0.05;70.1% vs -4.2%,P ＜ 0.05;66.9% vs 24.5%,P ＜ 0.05).Conclusion The rapid decrease of PD-1 expression on peripheral CD8+ T cells after antiviral treatment with ETV is positvely correlated with the decrease of serum HBV DNA loads and may be used as a predictive index for HBeAg seroconversion in HBeAg positive patients.     

胃癌患者CD4~+和CD8~+T细胞的PD-1高表达与肿瘤免疫逃逸的关系

目的探讨胃癌患者CD4~+和CD8~+T细胞的程序性死亡受体(PD)-1与肿瘤免疫逃逸的关系。方法采用流式细胞术检测胃癌患者外周血中CD4~+、CD8~+T细胞的PD-1表达情况;分析胃癌患者外周血单核细胞PD-L1表达情况;比较胃癌组织、胃癌患者外周血与健康人胃黏膜中CD4~+、CD8~+T细胞的PD-1表达情况;酶联免疫吸附(ELISA)分析胃癌组织中表达PD-1阳性与PD-1阴性的CD4~+、CD8~+T细胞因子分泌情况。结果胃癌患者外周血中CD4~+、CD8~+T细胞PD-1表达和单核细胞的PD-L1表达均显著高于对照组(P<0.05);胃癌组织中CD4~+、CD8~+T细胞表达的PD-1显著高于胃癌外周血和正常人胃黏膜中的PD-1表达;胃癌组织中PD-1阳性的CD4~+,CD8~+T细胞分泌的相关细胞因子干扰素(IFN)γ、白细胞介素(IL)-17、granzyme B,肿瘤坏死因子(TNF)显著低于PD-1阴性。结论胃癌患者体内CD4~+、CD8~+T细胞高表达PD-1抑制了T细胞的功能,促进胃癌的免疫逃逸。     

CYP1A1 , GSTM1 , GSTT1 and NQO1 polymorphisms and colorectal adenomas in Japanese men

AIM: To investigate the role of functional genetic poly-morphisms of metabolic enzymes of tobacco carcinogens in the development of colorectal adenomas. METHODS: The study subjects were 455 patients with colorectal adenomas and 1052 controls with no polyps who underwent total colonoscopy in a preretirement health examination at two Self Defense Forces hospitals. The genetic polymorphisms studied wereCYP1A1*2A (rs 4646903), CYP1A1*2C (rs 1048943), GSTM1 (null or non-null genotype), GSTT1 (null or non-null genotype) and NQO1 C609T (rs 1800566). Genotypes were determined by the polymerase chain reaction (PCR)-restriction fragment length polymorphism or PCR method using genomic DNA extracted from the buffy coat. Cigarette smoking and other life-style factors were ascertained by a self-administered questionnaire. The associations of the polymorphisms with colorectal adenomas were examined by means of OR and 95%CI, which were derived from logistic regression analysis. Statistical adjustment was made for smoking, alcohol use, body mass index and other factors. The gene-gene interaction and effect modification of smoking were evaluated by the likelihood ratio test. RESULTS: None of the five polymorphisms showed a significant association with colorectal adenomas, nor was the combination of GSTM1 and GSTT1 . A borderline significant interaction was observed for the combination of CYP1A1*2C and NQO1 (P = 0.051). The OR associated with CYP1A1*2C was significantly lower than unity among individuals with the NQO1 609CC genotype. The adjusted OR for the combination of the CYP1A1*2C allele and NQO1 609CC genotype was 0.61 (95%CI: 0.42-0.91). Although the interaction was not statistically significant (P = 0.24), the OR for individuals carrying the CYP1A1*2C allele and GSTT1 null genotype decreased significantly compared with those who had neither CYP1A1*2C allele nor GSTT1 null genotype (adjusted OR: 0.69, 95%CI: 0.49-0.97). Smoking did not modify the associations of the individual polymorphisms with colorectal adenomas. There w     

Cytochrome 1A1 and 1B1 gene diversity in the Zanzibar islands

Amodiaquine (AQ) is a 4‐aminoquinoline widely used in the treatment of malaria as part of the artemisinin combination therapy (ACT). AQ is metabolised towards its main metabolite desethylamodiaquine mainly by cytochrome P450 2C8 (CYP2C8). CYP1A1 and CYP1B1 play a minor role in the metabolism but they seem to be significantly involved in the formation of the short‐lived quinine‐imine. To complete the genetic variation picture of the main genes involved in AQ metabolism in the Zanzibar population, previously characterised for CYP2C8, we analysed in this study CYP1A1 and CYP1B1 main genetic polymorphisms. The results obtained show a low frequency of the CYP1A1*2B/C allele (2.4%) and a high frequency of CYP1B1*6 (approximately 42%) followed by CYP1B1*2 (approximately 27%) in Zanzibar islands. Genotype data for CYP1A1 and CYP1B1 show a low incidence of fast metabolisers, revealing a relatively safe genetic background in Zanzibar’s population regarding the appearance of adverse effects.     

1-Naphthyl isocyanate and 1-naphthylamine as metabolites of 1-naphthylisothiocyanate

Abstract: The importance of the bioactivation of 1-naphthylisothiocyanate was studied. Forty minutes after 1-naphthylisothiocyanate administration to rats, bile was collected over a 2.5-h period; the liver was then excised and homogenized. 1-naphthylisothiocyanate and its metabolites in bile and liver of rats were identified and quantified using coupled gas chromatography-mass spectrometry. Three main compounds were found in all 1-naphthylisothiocyanate-treated animals. They were identified as 1-naphthyl isocyanate, 1-naphthylamine and the parent compound, 1-naphthylisothiocyanate. When rats were given cycloheximide, which attenuates 1-naphthylisothiocyanate toxicity, 30 min before 1-naphthylisothiocyanate (300 mg/kg), 1-naphthyl isocyanate concentration was significantly lower than in rats receiving only 1-naphthylisothiocyanate. The appearance of 1-naphthylamine was also inhibited by cycloheximide, although not to the same extent as 1-naphthyl isocyanate. On the other hand, phenobarbital, which potentiates 1-naphthylisothiocyanate hepatotoxicity, enhanced 1-naphthyl isocyanate and 1-naphthylamine formation. It is suggested that 1-naphthyl isocyanate, 1-naphthylamine and the highly reactive sulfur released from 1-naphthylisothiocyanate might be involved in the hepatotoxic effect of 1-naphthylisothiocyanate.     

Interleukin-1 and interleukin-1 antagonism.

The polypeptide cytokine interleukin-1 (IL-1) affects nearly every tissue and organ system. IL-1 is the prototype of the pro-inflammatory cytokines in that it induces the expression of a variety of genes and the synthesis of several proteins that, in turn, induce acute and chronic inflammatory changes. IL-1 is also the prototypic "alarm" cytokine in that it brings about increases in a variety of defense mechanisms, particularly immunologic and hematologic responses. Most studies on the biology of IL-1 have been performed in animals, but human subjects have recently been injected with recombinant IL-1 and the results confirm the two fundamental properties of IL-1 as being both a mediator of disease as well as of host defense. However, in either situation, over or continued production of IL-1 leads to debilitation of normal host functions; therefore, reduction of IL-1 synthesis or its effects becomes a target of therapy in many diseases. In this review, the structure, gene expression, synthesis, and secretion of IL-1 are described. In addition, the two IL-1 surface receptors, possible signal transduction mechanisms, various biologic activities, and production of IL-1 during disease states are discussed. Similarities and differences between IL-1, tumor necrosis factor, and IL-6 are presented. Although various agents for reducing the synthesis and/or for antagonizing the effects of IL-1 have been proposed, the recent cloning of a naturally occurring IL-1 receptor antagonist (IL-1ra) has opened new experimental and clinical approaches. The ability of this IL-1ra to block the triggering of IL-1 receptors in animals without agonist effects has reduced the severity of diseases such as hemodynamic shock, lethal sepsis, inflammatory bowel disease, experimental arthritis, and the spontaneous proliferation of human leukemic cells.     

甲型H1N1与季节性H1N1流感病毒血凝素基因比较分析

目的分析泰安市2008～2009年度季节性流感与2009年度甲型H1N1流感病原学检测结果 ,比较季节性H1N1与甲型H1N1血凝素基因变异情况。方法选择国家级流感监测哨点医院以及暴发疫情的疫点,采集流感样病例的鼻咽拭子标本,通过RealtimePCR进行病毒检测,用MDCK细胞进行病毒分离,通过RT-PCR扩增血凝素HA1片段的基因并测序,利用生物信息学进行序列分析。结果 2008～2009年共检测鼻咽拭子标本283份,分离出流感病毒33株,分离阳性率为11.67%,其中季节性H1N1亚型31株。2009年5月1日～12月31日,检测鼻咽拭子标本996份,流感核酸检测阳性417份,阳性率为41.86%,其中甲型H1N1337份,季节性H1N1亚型1份。6株季节性H1N1病毒均在多个氨基酸位点上发生变异,与疫苗株A/Brisbane/59/2007(H1N1)比较,有11个位点发生了突变,其中5个位点位于抗原决定簇上;测序成功的6株甲型H1N1病毒在多个氨基酸位点发生变异,与疫苗株A/California/07/2009(H1N1)比较,有6个位点发生突变,其中1个位点位于抗原决定簇的B区。结论 2008～2009年度季节性H1N1为优势株,甲流暴发后,甲型H1N1成为绝对优势毒株。季节性H1N1分离株有多处氨基酸替换,抗原决定簇B区变异频繁;甲型H1N1病毒分离株的基因有变异,但关键位点第222位仍为D(天冬氨酸),与疫苗株相比抗原决定簇的关键位点变化不大。     

Trib1 and Evi1 cooperate with Hoxa and Meis1 in myeloid leukemogenesis

Cooperative activation of Meis1 and Hoxa9 perturbs myeloid differentiation and eventually leads myeloid progenitors to leukemia, yet it remains to be clarified what kinds of subsequent molecular processes are required for development of overt leukemia. To understand the molecular pathway in Hoxa9/Meis1-induced leukemogenesis, retroviral insertional mutagenesis was applied using retrovirus-mediated gene transfer. The mice that received Hoxa9/Meis1-transduced bone marrow cells developed acute myeloid leukemia (AML), and Trib1, Evi1, Ahi1, Raralpha, Pitpnb, and AK039950 were identified as candidate cooperative genes located near common retroviral integration sites. Trib1 and Evi1 were up-regulated due to retroviral insertions, and coexpression of these genes significantly accelerated the onset of Hoxa9/Meis1-induced AML, suggesting that Trib1 and Evi1 are the key collaborators. Furthermore, Trib1 by itself is a novel myeloid oncogene, enhancing phosphorylation of ERK, resulting in inhibition of apoptosis. These results demonstrate the importance of specific oncogene interaction in myeloid leukemogenesis.     

GSTT1, GSTM1 and GSTP1 polymorphisms and susceptibility to juvenile idiopathic arthritis

OBJECTIVE: In this study we have analyzed GSTM1, GSTT1 and GSTP1 polymorphisms in patients with juvenile idiopathic arthritis (JIA), to investigate a possible role of these genes as genetic components of the disease. METHODS: A total of 103 individuals (49 oligoarticular, 41 polyarticular and 13 systemic) were analyzed for the three polymorphisms, using a PCR/RFLP methodology. RESULTS: We have observed significantly increased frequencies of individuals with GSTT1 null genotype in JIA patients comparing to controls (37% x 21%; p=0.0183). There was a 2-fold increased risk (OR 2.2, 95% CI 1.2-4.1) associating the disease with the GSTT1 null genotype. Considering the subgroups (oligoarticular, polyarticular and systemic), the results indicated an association between polyarticular and systemic patients and the GSTT1 null genotype. There was a 2-fold increased risk for polyarticular patients (OR 2.4, 95%, CI 1.1-5.4), and a 4-fold increased risk for systemic patients (OR 4.4, 95%, 1.3-14.5). CONCLUSION: The GSTT1 null genotype seems to be involved in polyarticular and systemic JIA.     

Glutathione S-transferase M1, T1, and P1 genotypes and rheumatoid arthritis

OBJECTIVE: To determine the effects of genetic polymorphisms of glutathione S-transferase (GST) M1, GSTT1, and GSTP on risk and severity of rheumatoid arthritis (RA) in a Korean population. METHODS: A total of 258 patients with RA and 400 disease-free controls were enrolled. GST genotypes were determined by RFLP-PCR. HLA-DRB 1 typing and further subtyping of all alleles was performed using sequence-specific oligonucleotide probe hybridization after PCR. Severity of RA among cases was assessed by Steinbrocker anatomical stage. Risk was assessed by calculating the age and sex adjusted odds ratio (OR) and 95% confidence intervals (CI). RESULTS: The OR for risk of RA with the GSTM1-null genotype was 1.40 (95% CI 1.02- 1.92, p = 0.04), and 1.86 (95% CI 1.12- 3.09, p = 0.005) among individuals without the shared epitope (SE). Among patients with RA, the OR for risk of severe RA for the GSTM1-null genotype was 2.45 (95% CI 1.04- 5.77, p = 0.02). No association was observed between the GSTT1 or GSTP1 genotypes and either risk or severity of RA. CONCLUSION: These results suggest that the deletion polymorphism of GSTM1 is associated with increased susceptibility for RA, particularly among individuals who are not carriers of the HLA-DRB 1 SE.