结直肠癌人类白细胞抗原ABC表达与预后的相关性及干扰素γ的诱导作用研究

目的探讨人类白细胞抗原ABC（以下简称ABC抗原）在结直肠癌组织中的原位表达及其与生存预后之间的关系,揭示干扰素γ（IFN-γ）通过肿瘤抗原处理相关转运蛋白（TAP）途径对ABC抗原的诱导表达作用。方法采用免疫组织化学SP法检测62例结直肠癌组织中ABC抗原的表达,另采用RT-PCR方法分别检测癌组织中IFN-γ mRNA、TAP mRNA及ABC mRNA的含量。结果结直肠癌ABC抗原表达在年龄、性别、肿瘤大小及肿瘤所在部位中差异无统计学意义（χ^2值分别为0.38、0.78、2.70、0.52,均P〉0.05）,在Duke＇s分期、组织学分型、预后及IFN-γ表达中差异有统计学意义（χ^2值分别为8.26、4.11、4.32、4.96,均P〈0.05）;肿瘤组织中随着分化程度的降低,IFN-γ mRNA的含量降低,TAP mRNA及ABC mRNA也随之降低,三者具有协同效应。结论结直肠癌组织中ABC抗原的表达可以作为判断患者预后的指标之一,TAP是参与IFN-γ诱导肿瘤细胞增强表达ABC抗原过程的重要分子之一。     

Differential regulation of interleukin-6 receptors by interleukin-6 and interferons in multiple myeloma cell lines

Interleukin-6 (IL-6) mediates pleiotropic functions through specific receptors (IL-6R) composed of an 80-kDa binding protein, associated with a non-ligand binding protein (gp130) which transduces the signal. Because IL-6 is the major tumor growth factor in multiple myeloma, we investigated the regulation of IL-6R in two human multiple myeloma cell lines. Binding experiments with ¹²⁵I-labeled IL-6 showed that IL-6R were expressed at a high density on RPMI-8226 cells (15 000 receptors/cell), but no specific binding was detected on XG-1 cells, whose growth depends on the presence of exogenous IL-6. However, when IL-6 was removed from the culture medium, high-affinity IL-6R appeared on the surface of XG-1 cells (5300 sites/cell). Treatment of RPMI-8226 cells with IL-6 reduced the number of IL-6R without changing their affinity. This reduction was dose dependent and was not affected by acid treatment which dissociates ligand-receptor complexes. Cross-linking experiments showed that the formation of one IL-6/receptor complex of 160 kDa markedly decreased upon IL-6 treatment, while the other complex of 190 kDa became undetectable. These data provide evidence for ligand-induced down-regulation of membrane IL-6R expression in myeloma cells. Treatment of RPMI-8226 cells with interferon-α (IFN-α), which inhibits the growth of these cells, stimulated IL-6R expression and increased the formation of the 160-kDa IL-6/receptor complex. This stimulation was specific for IFN-α, since IFN-γ reduced the number of IL-6R. These data indicate that, in myeloma cells, IL-6R are differentially regulated by IL-6 and IFN-α.     

人IFN-λ1-3型mRNA实时荧光定量PCR检测方法的建立

【目的】建立人IFN‐λ1‐3型mRNA实时荧光定量聚合酶链式反应（qRT‐PCR）检测方法。【方法】根据人IFN‐λ1‐3型mRNA的基因序列合成特异性引物及探针,用体外转录方法制备IFN‐λ1‐3型的mRNA标准品,建立人IFN‐λ1‐3型mRNA qRT‐PCR检测方法。【结果】IFN‐λ1‐3型RNA标准品与对应的CT 值有良好的线性关系;扩增效率均在90％～100％之间;灵敏度分别是：1×100 co pies／μL、1×101 co pies／μL、1×101 co pies／μL ;变异系数均小于2％;各型间交叉反应不明显。【结论】本研究建立的IFN‐λ1‐3型mRNA qRT‐PCR检测方法灵敏度高,具有良好的重复性及特异性。     

慢性丙型肝炎复发合并亚临床甲状腺功能减退症1例

一、病例资料 患者女性,37岁,汉族,广东籍,教师.因"发现抗-HCV阳性5年"就诊.患者2003年体检时发现"抗-HCV阳性",因肝功能正常,未予以治疗.2004年8月起用普通干扰素联合利巴韦林治疗1年,HCV RNA转阴后停药,定期复查.停药6个月出现病毒学突破,肝功能异常,此后未进行抗病毒治疗,肝功能反复.2005年10月体检时发现"脂肪肝"及"甲状腺混合性结节",均未进行特殊治疗;无特殊个人吏、家族史.     

慢性乙型肝炎抗病毒疗效与外周血单个核细胞内乙型肝炎病毒DNA水平的关系

目的 探讨慢性乙型肝炎(CHB)抗病毒疗效与达到停药标准时外周血单个核细胞(PBMC)内HBV DNA水平的关系.方法 入选90例经抗病毒治疗达到停药标准的CHB患者,其中应用IFN 44例,应用核苷类药物46例.所有患者均于停药时检测PBMC内HBV DNA,比较阴性组和阳性组治疗前血清HBV DNA水平与达到停药标准时PBMC内HBV DNA的关系,观察停药时PBMC内HBV DNA水平与复发的关系.计量资料采用t检验,计数资料采用X2检验.结果 90例CHB患者停药时,PBMC内HBV DNA阴性组67例,阳性组23例.CHB患者血清HBV DNA阳转率在PBMC内HBV DNA阴性组为13.4%(9/67例),显著低于阳性组的73.9%(17/23例),差异有统计学意义(X2=30.4873,P<0.01).PBMC内HBV DNA阴性组与阳性组在肝病复发ALT升高幅度(t=0.8729,P=0.3913)、停药后复发时间(t=1.9222,P=0.0665)均差异无统计学意义,而在血清HBV DNA反弹幅度则差异有统计学意义(t=2.7493,P=0.0112).5例患者获得HBsAg血清学转换,且均未检测到PBMC内HBV DNA,随访6～12个月无一例复发.PBMC内HBV DNA阳性组治疗前血清HBV DNA水平为(7.2±1.1)lg拷贝/mL,显著高于阴性组的(5.2±2.1)lg拷贝/mL(t=4.3557,P<0.01).结论 经抗病毒治疗达到停药标准的CHB患者,其停药时的PBMC内HBV DNA水平可能是预测抗病毒疗效持久性的重要因素之一.

Abstract：

Objective To explore the relationship between the antiviral effect and peripheral blood mononuclear cell (PBMC) hepatitis B virus (HBV) DNA when the patients reach the standard of withdrawal of antiviral therapy in chronic hepatitis B (CHB).Methods Ninety CHB patients treated with interferon(n=44) or nucleot (s) ide(n=46) who reached the standard of withdrawal of antiviral therapy were recruited.HBV DNA levels in PBMCs were tested at the end of treatment,and its relationship with serum HBV DNA level before treatment in PBMC HBV DNA positive group and negative group were compared.The correlation between HBV DNA in PBMCs at the end of treatment and relapse were explored.Measurement data were analyzed by student t test and enumeration data were analyzed by X2 test.Results Among 90 patients,67(74.4%) were PBMC HBV DNA negative at the end of treatment,and 23(25.6%) were positive.The serum HBV DNA positive conversion rate in PBMC HBV DNA negative patients was 13.4%,which were significantly lower than that in positive group (73.9%) (X2=30. 4873, P<0.01 ). There were no significant differences of alanine aminotransferase (ALT) levels when hepatitis flare (t=0. 8729, P=0. 3913) and relapse time (t=1. 9222, P=0. 0665) between PBMC HBV DNA negative group and positive group after withdrawal of therapy, while the serum HBV DNA rebound was greater in positive group than that in negative group (t=2. 7493, P=0. 0112). There were five patients who achieved hepatitis B surface antigen (HBsAg) seroconversion, whose PBMC HBV DNA were all undetectable, and none relapsed during follow-up for 6-12 months. The pretreatment HBV DNA as level in PBMC HBV DNA positive was (7.2±1.1) lg copy/mL, which was much higher than that in negative group[(5.2±2.1) lg copy/mL] (t=4. 3557, P<0.01). Conclusions In patients who reach the standard of drug withdrawal,PBMC HBV DNA at the end of treatment is an important predictor for durability of antiviral therapy in CHB.     

干扰素诱导慢性乙型肝炎患者出现肝细胞损害敏感期的始动因素

目的 探讨干扰素诱导慢性乙型肝炎患者出现肝细胞损害敏感期的始动因素。方法 对44例乙型肝炎e抗原(HBeAg)和乙型肝炎病毒(HBV)DNA均阳性的慢性乙型肝炎患者进行干扰素治疗,定期检测血清丙氨酸氨基转移酶(ALT)以及外周血中乙型肝炎表面抗原(HBsAg)、HBeAg、抗-HBc和HBVDNA 含量,治疗前进行肝活体组织检查。结果 干扰素治疗后第6个月的HBeAg阴转率为75.0％,随访1年后为68.2％。肝细胞损害敏感期出现率为47.7%,其平均出现时间(3.14±1.49)周,平均持续时间(8.24±3.52)周,ALT平均升幅为基础ALT的(1.73±1.13)倍。肝细胞损害敏感期出现有利于提高干扰素治疗显效(Fisher精确概率,P—0.028)。治疗前外周血中HBeAg水平较高、肝内炎症活动度适中及HBcAg表达活跃有利于肝细胞损害敏感期的出现。从发生时间分析,外周血中的HBeAg和抗-HBc水平下降与肝细胞损害敏感期的起始相吻合。结论 肝细胞损害敏感期的始动因素可能为:干扰素治疗抑制了病毒蛋白的合成,导致外周血中HBeAg和抗-HBc水平快速下降,降低了对识别HBcAg的细胞毒性T淋巴细胞(CTL)的免疫封闭作用,从而激活了CTL细胞介导的乙型肝炎病毒感染肝细胞毒性作用。     

丙型肝炎病毒1b亚型E2区序列与干扰素治疗应答的关系

目的探讨上海地区丙型肝炎病毒(HCV)1b亚型感染者E2区的高变区1(HVR1)和高变区2(HVR2)、双链RNA激活的蛋白激酶真核翻译启动因子磷酸化同源区域(PePHD)及其邻近区域的基因序列与干扰素应答的相关性。方法收集上海地区24例HCV1b慢性感染者在α干扰素治疗前后及随访过程中的血清标本,用逆转录聚合酶链反应扩增E2区的HVR1、HVR2、PePHD及其邻近区域的基因并进行测序和氨基酸同源性分析。结果治疗前的HVR1区氨基酸序列高度变异,基于HVR1序列的种系发生树提示序列变化与干扰素应答类型无关。持续应答组(SR)、无应答组(NR)和复发组(TR)治疗前的HVR2区平均氨基酸变异数分别为2.0、1.0和1.5,统计学分析显示SR组与NR组差异有显著性(t=3.098,P<0.05)。治疗前的PePHD序列高度保守,无1例存在氨基酸变异。观察4例NR患者治疗过程中的PePHD序列,也未发现有任何氨基酸的突变。进一步比较PePHD邻近区域的氨基酸序列发现存在个别位点的变异(平均氨基酸变异SR组6.67,NR组5.75,TR组7.43),但变异的多少与疗效无关(SR组∶NR组t=1.413,P=0.195;SR组∶TR组t=1.706,P=1.107;TR组∶NR组t=0.682,P=0.504)。结论与HCVE2基因的HVR1区、PePHD区及其邻近区域序列与患者对干扰素的应答无相关性比较,HVR2区的序列变化可能与干扰素应答存在     

Effect of pegylated interferon alpha 2b plus ribavirin treatment on plasma transforming growth factor-β1, metalloproteinase-1, and tissue metalloproteinase inhibitor-1 in patients with chronic hepatitis C

AIM: To evaluate the effect of antiviral treatment on plasma levels of transforming growth factor-beta1 (TGF-beta1), metalloproteinase 1 (MMP-1), and tissue inhibitor of metalloproteinase-1 (TIMP-1) in patients with chronic hepatitis C.METHODS: TGF-beta1, MMP-1, and TIMP-1 plasma concentrations were measured by an enzyme immunoassay in 28 patients, during 48 wk of treatment with pegylated interferon-alpha 2b (PEG-IFN-alpha2b) plus ribavirin (RBV) and after 24 wk of follow-up. Patients were divided into two groups: responders (R) and non-responders (NR) related to achieved sustained virologic response. Normal values were evaluated in plasma samples of 13 healthy volunteers.RESULTS: Baseline plasma concentrations of TGF-beta1 and TIMP-1 (30.9+/-3.7 and 1 506+/-61 ng/mL respectively) measured in all subjects significantly exceeded the normal values (TGF-beta1: 18.3+/-1.6 ng/mL and TIMP-1: 1 102+/-67 ng/mL). In contrast, pretreatment MMP-1 mean level (6.5+/-0.9 ng/mL) was significantly lower than normal values (11.9+/-0.9 ng/mL). Response to the treatment was observed in 12 patients (43%). TGF-beta1 mean concentration measured during the treatment phase decreased to the control level in both groups. However at wk 72, values of NR patients increased and became significantly higher than in R group. TIMP-1 concentrations in R group decreased during the treatment to the level similar to normal. In NR group, TIMP-1 remained significantly elevated during treatment and follow-up phase and significant difference between both groups was demonstrated at wk 48 and 72. MMP-1 levels were significantly decreased in both groups at baseline. Treatment caused rise of its concentration only in the R group, whereas values in NR group remained on the level similar to baseline. Statistically significant difference between groups was noted at wk 48 and 72.CONCLUSION: These findings support the usefulness of TGF-beta1, TIMP-1, and MMP-1 in the management of chronic hepatitis C. Elevated TIMP-1 and low MMP-1 plasma concentrations during antiviral therapy may indicate medication failure.     

Ⅲ型干扰素表达的调控机制

目前,人们对Ⅲ型IFN(IFNλ)的功能和表达调控机制的了解还不完全.Ⅲ型IFN的表达方式和生物学效应与Ⅰ型IFN非常相似,所以很有可能其调控表达的机制也与Ⅰ型IFN相同.虽然最开始人们普遍认为Ⅰ型IFN只有在病毒感染的情况下才会表达,但是现在越来越多的研究指出胞内的细菌和寄生虫也可以促进其表达.Ⅰ型和Ⅲ型IFN调控机...     

HBV DNA及HBeAg定量在乙型肝炎患者抗病毒治疗方案选择中的指导作用

目的 根据慢性乙型肝炎患者HBV DNA及HBeAg定量高低的不同组合,观察其对IFN、阿德福韦酯(ADV)的治疗效果,寻求抗病毒药物的最佳治疗方案.方法 选择HBeAg阳性患者165例,根据所测HBV DNA及HBeAg定量高低的不同,将其分为A、B、C、D组.每组患者再随机分为IFN治疗组及ADV治疗组,48周时观察不同药物在各组患者治疗效果中的各项指标.结果 IFN治疗组对HBV DNA抑制率、HBV DNA水平下降(≥2lg拷贝/ml)的患者比例、HBV DNA阴转率及ALT复常率与ADV治疗组比较差异均无统计学意义,但在HBeAg定量下降≥500.00 COL/ml的患者比例、HBeAg阴转率、HBeAg血清转换率等项指标比较差异有统计学意义(P<0.05).并提示随着HBV DNA及HBeAg定量的增高,IFN和ADV的疗效依次下降,表现在ADV治疗组更为明显,显示在HBeAg血清转换率方面,IFN疗效明显优于ADV.结论 根据HBV DNA及HBeAg定量的高低,对选择不同的抗病毒治疗方案具有指导作用.