Enhanced complement-susceptibility and dysfunction of lymphocytes in paroxysmal nocturnal haemoglobinuria (PNH)

We investigated the complement-susceptibility of paroxysmal nocturnal haemoglobinuria (PNH) lymphocytes in relation to their dysfunction. When assessed by complement-mediated lysis induced by monoclonal antibodies (CD5 or CD20) and rabbit complement, the complement-susceptibility of lymphocytes from patients with PNH, both CD5+ (T cells) and CD20+ (B cells), was greater than that from controls (P less than 0.001). This susceptibility was further enhanced, both in normal controls (P less than 0.01) and in patients with PNH (P less than 0.001), when the activity of decay-accelerating factor (DAF) on the lymphocytes was blocked with anti-DAF monoclonal antibody. DAF amounts in mononuclear cells (MNC) from patients with PNH, measured by an enzyme-linked immunosorbent assay (ELISA), were lower than those of the controls (P less than 0.001). The expressions of DAF on T cells and on B cells from patients with PNH were significantly decreased (P less than 0.05 in T cells: P less than 0.01 in B cells). MNC from patients with PNH responded less to phytohaemagglutinin (PHA) and concanavalin A (Con A) than MNC from the controls (P less than 0.001). In contrast, the responses of PNH MNC to poke weed mitogen (PWM) or lipopolysaccharide (LPS) were not impaired. MNC from a normal donor preincubated with anti-DAF became less responsive to PHA or Con A. We conclude that PNH lymphocytes show enhanced complement-susceptibility and that they are involved in their own expression of DAF as well as in other types of peripheral blood cells. The results of the responses to various lectins suggest the dysfunction of T cells in PNH.     

Decreased interleukin-2 receptor β chain expression by peripheral blood lymphocytes in chronic liver disease

To investigate the decrease in natural killer (NK) activity in chronic liver disease, interleukin-2 receptor beta chain (IL-2R beta) expression was assessed by peripheral blood lymphocytes (PBL) using flow cytometry and an IL-2R beta chain-specific mouse monoclonal antibody. The percentage of IL-2R beta chain-positive PBL was significantly decreased in patients with chronic viral hepatitis, liver cirrhosis and hepatocellular carcinoma in comparison with normal controls (P less than 0.01). Among chronic viral hepatitis patients, it was significantly less in those with chronic active hepatitis than in those with chronic persistent hepatitis (P less than 0.05). Two-colour flow cytometry revealed that the IL-2R beta chain was mainly expressed by CD8+ or CD16+ cells in both the controls and the liver disease patients. CD8dull+ cells (NK cells) constituted more than 60% of the CD8+ cells expressing the IL-2R beta chain. Expression of the IL-2R beta chain with CD8 or CD16 was also significantly decreased in chronic liver disease patients compared with controls. In chronic viral hepatitis, there was a significant correlation between NK activity and the percentage of IL-2R beta+ PBL (P less than 0.001, r = 0.916), as well as between NK activity and the percentage of PBL co-expressing both the IL-2R beta chain and CD16 (P less than 0.001, r = 0.850). These findings suggest that decreased expression of the IL-2R beta chain by PBL may result in diminished NK activity in chronic liver disease.     

乙型肝炎病毒相关肝功能衰竭合并侵袭性肺曲霉菌病的临床特点与转归

目的 探讨HBV相关肝功能衰竭合并侵袭性肺曲霉菌病(IPA)的临床特征、危险因素、诊治及预后.方法 采用病例对照研究的方法,回顾性分析43例HBV相关肝功能衰竭合并IPA患者的临床特征、危险因素、诊治经过、预后及转归等,并选取同期住院的43例HBV相关肝功能衰竭合并单纯细菌感染患者以及43例未合并感染的HBV相关肝功能衰竭患者作为对照.计量资料采用方差分析,计数资料采用卡方检验,多因素分析采用Logistic回归分析.结果 合并感染的HBV相关肝功能衰竭患者PTA和PLT显著下降,MELD评分升高,尤以IPA组患者为著,合并IPA前后的对比结果亦是如此.IPA组患者1年内病死率为100.0%,明显高于单纯细菌感染组的65.10%,两者均高于未合并感染组的39.50%;Logistic回归分析结果显示,住院时间延长、使用大量抗菌药物、侵入性操作频繁等是HBV相关肝功能衰竭合并IPA的危险因素.结论 HBV相关肝功能衰竭患者合并IPA后病情迅速恶化、病死率高、预后差,提高对其早期表现的认知度,尽早诊断和进行强有力的抗真菌治疗,是提高HBV相关肝功能衰竭合并IPA患者生存率的有效方法.

Abstract：

Objective To investigate the clinical features, risk factors, diagnose, treatments and prognosis of patients with hepatitis B virus (HBV)-related liver failure complicated with invasive pulmonary aspergillosis (IPA). Methods A case-control study was performed to retrospectively analyze the clinical data of 43 patients with HBV-related liver failure complicated with IPA. A cohort of 43 patients with HBV-related liver failure complicated with bacterial infection only was analyzed at the same time and another cohort of 43 patients with HBV-related liver failure without any infections served as the control group. Results Compared with the control group, patients with infection had lower platelet counts and prothrombin activity, higher scores of model for end-stage liver disease (MELD), especially the patients with IPA (F=42.43,13.69,14, 22, all P＜0.01). And the similar results were observed in patients with IPA group before and after Aspergillus infection (F= 12.09,14.52,-16.74, all P＜0.01). Furthermore, the annual mortality was 100. 0% in patients with IPA,which was higher than that of patients complicated with bacterial infection only; and both groups were higher than control group of patients without infection. The statistic results showed that patients with longer length of stay, more antibiotics usage, and more invasive procedures were all risk factors of HBV-related liver failure complicated with IPA. Conclusions Conditions of patients with HBV-related liver failure deteriorate rapidly after complicated with IPA and patients have higher mortality and poorer prognosis.Therefore, it will be effective to enhance the recognition of the early clinical manifestations, and make diagnosis timely and treat with potent anti-fungal therapy to improve their survival.     

Effect of pyruvate dehydrogenase on production of interleukin-6 in peripheral blood mononuclear cells from patients with primary biliary cirrhosis

We investigated the effect of pyruvate dehydrogenase (PDH), a mitochondrial autoantigen, on the production of interleukin-6 (IL-6) by peripheral blood mononuclear cells (PBMC) from nine patients with primary biliary cirrhosis (PBC) and nine patients with other chronic liver disease (CLD) as a control. IL-6 activity was measured by the bioassay using MH-60 BSF2 cells. The mean level of spontaneous secretion of IL-6 from PBMC of PBC patients was significantly (P < 0.01) higher than that of CLD patients. In addition, the level of IL-6 production by PBMC of PBC patients with stimulation of PDH was significantly higher (P < 0.01) than that of CLD patients. On the other hand, the stimulation of PBMC from PBC and CLD patients by pokeweed mitogen (PWM) enhanced the IL-6 production. However, there was no significant difference in the levels of IL-6 produced by PBMC with stimulation of PWM + PDH between PBC and CLD. These findings suggest that PDH is involved in the production of large amounts of IgM and autoantibodies in PBC, by stimulating the production of IL-6 from mononuclear cells.     

Hepatocyte hepatitis B surface antigen expression in chronic hepatitis B virus carriers in Singapore: Correlation with viral replication and liver pathology

The hepatocyte hepatitis B surface antigen (HBsAg) expression in 149 liver biopsies from 124 chronic hepatitis B virus (HBV) carriers was correlated with serum HBV DNA status and histologic activity. Hepatocyte HBsAg was stained by the peroxidase-antiperoxidase method and serum HBV DNA was determined by dot blot hybridization. Sixty-five biopsies (44%) showed minimal changes (MC), 82 biopsies (55%) showed chronic liver disease (CLD) and 2 biopsies (1%) showed hepatocellular carcinoma. Hepatocyte HBsAg was found in 144 biopsies (97%). It was present in the cytoplasm of 141 specimens (95%) and/or plasma membrane of 48 specimens (32%). Approximately half (45%) of the cytoplasmic HBsAg-positive biopsies showed discrete distribution, while the other half (55%) were grouped. Fifty-five per cent (77 of 141) of cytoplasmic HbsAg-positive biopsies had CLD, while 44% (62 of 141) showed MC. There was no relationship between the presence of cytoplasmic HBsAg or its topographic distribution with disease activity. Membrane HBsAg distribution was similar for both groups of patients (MC vs CLD: 25 of 65 (38%) vs 23 of 82 (28%); P = NS). Serum HBV DNA was detected in 98 patients (66%) and was seen mostly in association with CLD (CLD vs MC: 61% vs 39%, P less than 0.001). It was also detected more often in the sera of patients with membrane HBsAg than in those with cytoplasmic HBsAg staining (41 of 48 (85%) vs 97 of 141 (67%); P less than 0.02). However, discrete distribution of cytoplasmic HBsAg was associated with positive serum HBV DNA when compared with grouped distribution (52 of 63 (83%) vs 43 of 78 vs (55%); P less than 0.005).     

乙型肝炎慢加急性肝功能衰竭患者免疫功能抑制与疾病严重程度的研究

目的 探讨乙型肝炎慢加急性肝功能衰竭(ACLF)患者体内免疫功能抑制与疾病严重程度的关系.方法 收集上海公共卫生临床中心2009年8月至2010年4月住院治疗的乙型肝炎ACLF患者27例(ACLF组),活动性慢性乙型肝炎患者28例(CHB组)和健康志愿者8名(对照组)的临床资料及外周血标本.以APACHEⅢ评分及肝性脑病程度作为反映疾病严重程度的量化指标.应用流式细胞术检测患者外周血中T淋巴细胞亚群的绝对计数和单核细胞表面人类白细胞抗原(HLA)-DR的表达量.采用CBA试剂盒中检测患者血浆中促炎性细胞因子及抑炎性细胞因子[干扰素(IFN)-γ、肿瘤坏死因子(TNF)-α、白细胞介素(IL)-2、IL-4、IL-10]的水平.采用SPSS 16.0软件进行数据统计.结果 与CHB组和对照组相比,ACLF组患者抑炎性细胞因子IL-10含量显著增加(Z=-4.279,U=124,P＜0.01;Z=-3.871,U=9.5,P=0.0001),促炎性因子IFN-γ、TNF-α、IL-2、IL-4则处于检测值下限;外周血中单核细胞表面HLA-DR表达量显著下调(Z=-4.714,U=98,P＜0.01;Z=-4.086,U=4,P＜0.01),HLA-DR表达量和APACHEⅢ评分存在显著负相关(R2=0.2667,P=0.0167);适应性免疫细胞CD4+T淋巴细胞绝对数量减少(Z=-4.411,U=116,P＜0.01;Z=-3.575,U=17,P=0.004).结论 乙型肝炎ACLF患者的免疫系统存在单核细胞功能抑制、CD4+T细胞耗竭及高含量抑炎性细胞因子的免疫紊乱状态,单核细胞HLA-DR表达量持续下降是反映疾病严重程度的指标.

Abstract：

Objective To explore the association of immune suppression with the severity of HBV-related acute-on-chronic liver failure(ACLF) patients.Methods From August 2009 to April 2010 in Shanghai Public Health Clinical Center, the peripheral blood samples and clinical data of 27 HBV-related ACLF patients (ACLF group), 28 patients wit h chronic active hepatitis B (CHB group)and 8 healthy individuals (Control group) were collected.APACHE Ⅲ score and the grade of hepatic encephalopathy were as quantitative index to evaluate the severity degree of the disease.The absolute counts of the subsets of T lymphocytes and human leukocyte antigen (HLA)-DR expression on the surface of monocyte in patients' peripheral blood were examined by flow cytometry, the proinflammatory cytokines and anti-inflammatory cytokines(IFN-γ, TNF-α, IL-2, IL-4, IL-10) in patients' plasma were detected by cytometric bead array (CBA) kit.The data was analyzed with SPSS 16.0 software.Results Compared with CHB group and control group, the level of anti-inflammatory eytokire IL-10 markedly increased ir HBV-related ACLF patients (Z= -4.279 ,U= 124, P＜0.01;Z= - 3.871, U= 9.5, P= 0.0001 ), however the level of pro-inflammatory cytokines IFN-γ、 TFN-α、IL-2 、 IL-4 in plasma were at low limit of detectable value.Meanwhile the expression quantity of HLA-DR on the peripheral blood monocytes significantly down-regulated (Z= -4.714, U= 98, P＜0.01;7= - 4.086, U = 4, P＜ 0.01), and there was negative correlation between HLA-DR expression quantity and APACHE Ⅲ score (R2 =0.2667, P=0.0167).In addition, the absolute counts of CD4+T lymphocytes in adaptive immune cells significantly decreased (Z= -4.411, U= 116, P＜0.01; Z=-3.575, U= 17, P= 0.0004).Conclusions The immune system of HBV-related ACLF patients displays immune dysfunction like monocyte function inhibition; CD4+ T lymphocytes depletion and high level of anti-inflammatory eytokines, the persistent down-regulation of the HLA-DR expression on monocyte is an indicator for the severity of disease.     

白细胞介素-17肝内表达与慢性乙型肝炎病毒感染所致肝纤维化的相关性

目的 探讨IL-17肝内表达与慢性HBV感染所致肝纤维化的相关性.方法 免疫组织化学法测定30例慢性HBV携带者、55例慢性乙型肝炎患者、20例乙型肝炎肝硬化患者肝组织内不同炎症程度分级和肝纤维化分期中IL-17的表达,ELISA法测定血清IL-17及肝纤维化指标HA、LN、PCⅢ、ⅣC的水平.组间差异性检验采用Kruskal-Wallis检验和Mann-Whitney检验,相关性分析采用Spearman分析.结果 肝组织IL-17表达水平在肝硬化组高于慢性乙型肝炎组(x2=25.3982,P=0.004),在慢性乙型肝炎组高于慢性HBV携带组(x2=11.5056,P=0.001);不同炎症程度及纤维化分级与IL-17表达强度呈正相关(r=0.718、0.693,均P＜0.01);肝组织IL-17主要集中于汇管区,其表达强度与血清HA、LN、PCⅢ、ⅣC呈正相关(r=0.793、0.834、0.722、0.883,均P＜0.01).结论 IL-17的肝内表达与肝内炎症程度分级及肝纤维化程度密切相关.     

Serum levels of macrophage migration inhibitory factor, Interleukin-17 and Interleukin-10 in patients with HBV-related liver disease

Objective To study the potential role of macrophage migration inhibitory factor(MIF),Interleukin-17(IL-17) and Interleukin-10(IL-10) in the development of HBV-related liver disease.Methods 48 patients with chronic hepatitis B(HBeAg negative and positive,24 cases;21cases of HBV-DNA negative and 27 cases of HBV-DNA positive),81 cases of hepatitis B patients with decompensated cirrhosis and 48 cases of primary liver cancer patients were collected as the experimental group,26 healthy people were as control group.Serum MIF,IL-17 and IL-10 were measured.Results MIF and IL-17 significantly increased,IL-10 significantly decreased in experimental group,compared with the control group(P<0.05),there was significant difference.In addition,there was no significant difference(P>0.05) between positive and negative of chronic hepatitis B.MIF,IL-17 and ALT levels were positively correlated(r=0.693,P<0.01;r=0.897,P<0.001),IL-10 and ALT was negatively correlated(r =-0.285,P=0.037).Conclusion These results indicated that MIF,IL-17 and IL-10 may participate in the pathological process of HBV-related liver disease,serum levels of MIF,IL-17 and IL-10 appear to reflect the severity of tissue injury in HBV-related liver disease.     

Cytokine profile in Egyptian hepatitis C virus genotype-4 in relation to liver disease progression

AIM: To observe the imbalance between T helper cell Th1 and Th2 cytokines in several chronic hepatitis disease at different stages of disease progression. METHODS: We measured the cytokine levels of Thl (IL-2 and IL-2R), Th2 (IL-10) and the pro-inflammatory cytokines (IL-6 and IL-6R and TNF and TNF-RI and Ⅱ) by the ELISA technique in the sera of 33 hepatocellular carcinoma (HCC) patients and 20 chronic liver disease (CLD) patients. In addition, 20 asymptomatic hepatitis C virus carriers and 20 healthy subjects negative for hepatitis C virus(HCV) markers served as controls. RESULTS: Anti-HCV antibodies were found to be positive in 94% of HCC cases and 75% of CLD cases. On the other hand, HCV viremia was detected using RT-PCR in 67% of HCC cases and 65% of CLD cases. HBsAg was positive in 9% of HCC cases and 30% of CLD cases. Also bilharzial-Ab was positive in 55% of HCC cases, 65% of CLD cases and in 70% of asymptomatic carriers (ASC). HCC patients had significantly higher values of IL-2R, TNF-RⅡ (P<0.001), and TNF-RI (P>0.05), but lower TNFα (P<0.001) and IL-6 (P = 0.032) in comparison to ASC. But, in comparison to non-cancer controls, HCC patients had higher values of IL-2R, IL-6R, TNF-RI and TNF-RⅡ, but lower TNF-α (P<0.001). CLD patients had higher IL-2R, TNF-RI, and TNF-RⅡ (P<0.001) than ASC. But, in comparison to non-cancer controls, CLD patients had higher values of IL-2R, TNF-RI and TNF- RⅡ, but lower TNF-α (P<0.001). IL-10 was higher (though not significantly) in HCC and CLD patients than in symptomatic carriers and non-cancer controls. CONCLUSION: Liver disease progression from CLD to HCC due to HCV genotype-4 infection is associated with an imbalance between Thl and Th2 cytokines. IL-2R, TNF-RI, and TNF-RⅡ could be used as potential markers.     

乙型肝炎肝硬化患者外周血调节性T细胞频率及血清IL-1β、IL-6和IL-10水平的变化

目的检测未抗病毒治疗的乙型肝炎肝硬化患者外周血调节性T细胞(Treg细胞)及其亚群频率和血清IL-1β、IL-6、IL-10水平,以探讨乙型肝炎肝硬化患者免疫功能的变化。方法在20例健康对照人群、17例慢性乙型肝炎(CHB)患者和38例乙型肝炎肝硬化(LC)患者,采用流式细胞仪检测外周血CD4+CD25+Foxp3(+Treg)细胞频率、CD4+CD39+Foxp3(+CD39+Treg)细胞和CD4+CTLA-4+Foxp3(+CTLA-4+Treg)细胞频率;采用ELISA法检测血清IL-1β、IL-6和IL-10水平。结果 LC患者Treg细胞频率、CD39+Treg细胞频率、CTLA-4+Treg细胞频率、血清IL-1β、IL-6和IL-10水平均高于正常人(P均<0.01)和CHB患者(P<0.05或P<0.01);失代偿期LC患者Treg细胞频率、CD39+Treg细胞频率、IL-6水平高于代偿期患者(P均<0.01);LC患者Treg细胞频率与CD39+Treg细胞频率(r=0.474,P<0.01)、CTLA-4+Treg细胞频率均呈正相关(r=0.330,P<0.05),Treg细胞频率(r=0.381,P<0.05)、CD39+Treg细胞频率(r=0.333,P<0.05)与TBil呈正相关,Treg细胞频率(r=-0.549,P<0.01)、CD39+Treg细胞频率(r=-0.437,P<0.01)、CTLA-4+Treg细胞频率(r=-0.368,P<0.01)与PTA呈负相关,CD39+Treg细胞频率与AST呈正相关(r=0.406,P<0.05)。结论 Treg细胞及其亚群频率和IL-1β、IL-6、IL-10水平的变化可作为临床初始治疗的乙型肝炎肝硬化患者辅助检查指标之一。     

血浆置换联合血液滤过对乙型肝炎相关慢加急性肝衰竭患者血清IL-17和IL-6的影响

目的探讨血浆置换联合血液滤过对乙型肝炎相关肝衰竭血清白细胞介素(IL)-17、IL-6的影响。方法 30例乙型肝炎慢加急性肝衰竭患者在内科治疗的基础上采用血浆置换联合血液滤过单次治疗。用ELISA检测各组血清IL-17、IL-6浓度,同时记录血清ALT、TBil等值并进行统计分析。结果肝衰竭组患者血清IL-17、IL-6水平分别为(123.5±23.0)pg/ml、(110.0±18.5)pg/ml,高于慢性乙型肝炎患者(48.5±6.3)pg/ml、(27.8±5.9)pg/ml和正常对照组(34.7±3.3)pg/ml、(12.1±5.1)pg/ml,P均<0.001;治疗后血清IL-17、IL-6水平分别为(84.7±21.4)pg/ml、(75.8±16.6)pg/ml,较治疗前下降(t=35.1,P<0.001;t=33.4,P<0.001);与好转组相比,人工肝对恶化组血清IL-17、IL-6清除效率降低(t=3.8,P<0.05;t=3.9,P<0.05);人工肝对血清IL-17、IL-6清除效率均与MELD评分呈负相关(r=-0.53、P=0.003;r=-0.43,P=0.015)。结论血浆置换联合血液滤过能有效降低肝衰竭患者血清IL-17、IL-6水平,其对IL-17、IL-6的清除效率可能与患者预后有关。     

Acute, hepatitis-A super-infection in HBV carriers, or chronic liver disease related to HBV or HCV

The impact of acute super-infection with hepatitis A virus (HAV) was determined in 20 asymptomatic carriers of the surface antigen (HBsAg) of hepatitis B virus (HBV), eight patients with HBV-related chronic liver disease (CLD), and four patients with CLD related to hepatitis C virus (HCV). For comparison, 100 patients with isolated HAV infection were also studied. The HBsAg carriers and patients with CLD related to HBV or HCV were significantly older than the patients with isolated HAV infection, with mean (S.D.) ages of 43.9 (14.1), 46.4 (16.0), 52.5 (8.6) and 28.4 (10.7) years, respectively (P < or = 0.02). There were no significant between-group differences in the baseline serum concentrations of alanine aminotransferase. All the patients with isolated HAV infection fully recovered. Fulminant or submassive hepatitis occurred in 11 (55%) of the HBsAg carriers and four (33%) of the 12 patients with CLD related to either HBV or HCV. Nine of the 15 patients with severe hepatitis died and the mortality rate among the HBsAg carriers was not significantly different from that among the CLD patients (25% v. 33%; P = 0.15). These fatal cases were all aged > 50 years and were significantly older [59.0 (2.1) years] than the six severe cases who recovered [43.2 (10.7) years] as well as the remaining 17 uncomplicated cases with CLD or HBsAg [40.3 (13.0) years] (P < or = 0.001). The results indicate that acute HAV is rarely fatal in young adults but may be severe and potentially fatal in patients with underlying chronic HBV or HCV infection, especially among the elderly. Vaccination against HAV should be considered for the patients at high risk who are negative for anti-HAV.     

Close association between basal cell layer antibodies and hepatitis B virus-associated chronic delta infection

Antibodies reacting in immunofluorescence with the basal cell layer of rat forestomach (BCLA) have been detected in 36 of 121 (30%) hepatitis B virus (HBV)-mediated chronic liver disease (CLD), in 1 of 30 (3%) HBV-negative CLD, in 3 of 36 (8%) alcoholic liver disease (with no correlation with serum HBV markers), in 1 of 25 (4%) primary biliary cirrhosis, in none of 19 HBV-related HBsAg-negative CLD and 60 healthy blood donors. Of 352 hospitalized patients with miscellaneous diseases (including immunological conditions), the antibodies were found in four (1%). In the 36 BCLA positive cases from HBV-mediated CLD, evidence of chronic delta infection was found in 34. The overall prevalence of BCLA in 68 delta cases was 50% (58% in chronic active hepatitis, 46% in cirrhosis), and in 28 delta negative cases was 4% (p less than 0.00002). BCLA of delta cases were mainly of the IgG class (38 of 41 sera), and high titers (up to 40,960) were found in the majority (66% greater than or equal to 1:640). The high titer BCLA has to be considered a marker of chronic delta infection in HBV cases.     

Anti-HCV positive hepatocellular carcinoma in cirrhosis : Prevalence, risk factors and clinical features

Recent reports indicate that hepatitis C virus (HCV) may play a role in the pathogenesis of hepatocellular carcinoma in cirrhotics. Using an ELISA test, we evaluated the prevalence of anti-HCV antibodies in 97 patients with hepatocellular carcinoma (HCC) in cirrhosis and in a group of 223 patients, including: 49 patients with HBsAg-positive chronic liver disease (CLD), 42 with alcoholic CLD, 110 with cryptogenic CLD and 22 with post-transfusional HBsAg-negative CLD. All diagnoses were histologically confirmed. Overall, anti-HCV-positive HCC were 64% of the total, with no statistically significant difference with respect to CLD (60.9%). The prevalence of anti-HCV was higher in cryptogenic HCC (80%) than in HBsAg-positive (60%) or alcoholic HCC (42.8%) (p less than 0.005). When HCC and cirrhosis of similar putative etiology were considered, anti-HCV prevalence was significantly higher in HCC than in cirrhosis only in the groups of patients with alcoholic liver damage (60% in HCC vs. 38% in cirrhosis, p less than 0.005). In HBsAg-positive patients, anti-HCV prevalence was twice as high in HCC than in CLD, but the difference was not statistically significant. Overall, anti-HCV prevalence in HCC was significantly higher than in alcoholic or HBsAg-positive CLD (p less than 0.001 and p less than 0.01, respectively) but lower than in cryptogenic CLD (p less than 0.001). Association between anti-HCV and anti-HBc was significantly more prevalent in patients with CLD than in those with HCC.(ABSTRACT TRUNCATED AT 250 WORDS)     

Serum antibodies to thymus epithelial cells in non-A, non-B and cryptogenic chronic liver disease

Antibodies against thymus epithelial cells (anti-TEC) and the basal cell layer (BCLA) of squamous epithelia have been described in association with HDV-related chronic liver disease (CLD). Data are lacking on their presence during nAnB virus infection. Sera from 51 patients with nAnB post-transfusion hepatitis, including acute and chronic cases diagnosed during a prospective study on candidates for cardiac surgery, and 167 with various forms of CLD were tested for the presence of anti-TEC and BCLA using indirect immunofluorescence on human thymus and rat forestomach sections. Both antibodies mainly occurred in nAnB, HDV and cryptogenic CLD (anti-TEC: 51%, 47% and 42%; BCLA: 29%, 38% and 31%, respectively). The prevalence of anti-TEC in nAnB CLD turned out to be higher than that recorded in alcoholic, HBV-related, autoimmune, liver and kidney microsomal antibody positive CLD and primary biliary cirrhosis (p ranging from less than 0.03 to less than 0.0004). Two monoclonal antibodies (Mabs) to cytokeratins gave a pattern superimposable on that of spontaneous anti-TEC (both Mabs) and BCLA (only one). Antibodies against epithelial constituents, presumably targeting cytokeratin-associated antigens, occur not only in HDV CLD, as previously reported, but also in nAnB CLD, where they might represent a diagnostic aid, due to the unavailability of reliable serological markers of nAnB infection. The close similarity of anti-TEC and BCLA status between nAnB and cryptogenic CLD suggests a nAnB etiology of at least a proportion of chronic liver patients at present scored as cryptogenic.     

Overlap and discrepancy between tests for anti-C100, anti-GOR and anti-CP9 in patients with chronic liver disease and inhabitants in Saga, Japan.

The authors evaluated the clinical significance of anti-C100, anti-GOR and anti-CP9 in hepatitis C virus (HCV)-related liver disease in two populations: 459 healthy subjects and 385 patients with chronic liver disease (CLD). Previously we reported high rates of mortality and morbidity (5.3%) of CLD in subjects in Saga, Japan. This was ascribed to the high prevalence (10.8%) of anti-HCV among randomized populations, as detected by the C100 ELISA test system, as compared with a finding of 2-3% in Japanese blood donors in the same decade. The incidence of anti-C100, anti-GOR and anti-CP9 detected by ELISA test system in the healthy population currently surveyed was 17.0%, 19.2% and 32.0% respectively, as compared with 75.3%, 60.3% and 73.0% respectively, in those with CLD. The incidence of positivity for at least one of the three antibodies was high (36.4%) among healthy subjects, and even higher (86.5%) among the patients with CLD. In the healthy subjects, incidence of positivity increased with age. The healthy and CLD populations differed in the proportion of cases positive for all three antibodies vs. those positive for at least one antibody: healthy subjects, 52/167, 31.1%, vs. CLD patients, 197/333, 59.2%; P less than 0.01. Among the anti-C100-positive healthy cases, these was a significantly high level of AST, ALT, ZTT and gamma GTP compared with negative cases, with or without anti-GOR and anti-CP9 (P less than 0.01-0.05). These observations suggest that the presence of anti-C100 may be related to the active state of HCV-related liver disease.(ABSTRACT TRUNCATED AT 250 WORDS)     

INCREASED IL-1 RECEPTOR ANTAGONIST (IL-lra) PRODUCTION AND DECREASED IL-l{beta}/IL-lra RATIO IN MONONUCLEAR CELLS FROM RHEUMATOID ARTHRITIS PATIENTS

In order to investigate the possible role of IL-1 receptor antagonist(IL-lra) in patients with rheumatoid arthritis (RA), this studywas undertaken to measure the amounts of IL-lra and interleukin-1ß(IL-1ß) protein produced by mononuclear cells (MNC)and to investigate the relationship between production of thesecytokines and clinical parameters. The MNC were cultured for24 h and the supernatants were measured for IL-lra and IL-1ßby ELISA kits. MNC from peripheral blood (PB) and synovial fluidof RA patients produced significantly higher amounts of IL-lrathan normal PBMNC (P < 0.01 and P < 0.05, respectively).When the IL-1ß/IL-lra ratio was calculated, IL-1ß/IL-lraratios of RA PBMNC were significantly lower than those of normalPBMNC (P < 0.001). The IL-1ß/IL-lra ratio of RAPBMNC was significantly higher in active RA patients than inRA patients in remission (P < 0.02). The amounts of IL-lraproduced by stimulated RA PBMNC positively correlated with thejoint score (P < 0.05), serum CRP levels (P < 0.05) andthe amounts of IL-1ß produced (P < 0.01). The amountsof IL-lra produced by unstimulated RA PBMNC did not correlatewith any of the clinical parameters studied. Gold sodium thiomalate(GST), but not auranofin, increased IL-lra production in vitro.These data suggest that increased IL-lra production in RA MNCis associated with IL-1ß produced, thus protectingfrom IL-1ß-mediated immune responses, and that enhancedIL-lra production by MNC cultured with GST is one of the mechanismsby which GST shows clinical efficacy. KEY WORDS: Rheumatoid arthritis, Interleukin-1 receptor antagonist, Mononuclear cells, Gold sodium thiomalate, Synovial fluid.     

Th17/Treg细胞因子在乙型肝炎慢加急性肝衰竭患者中的表达及意义

目的探讨乙型肝炎慢加急性肝衰竭患者(HBV-ACLF)血清中Th17/Treg细胞因子(IL-17/IL-35)的表达及其临床意义。方法收集兰州市第二人民医院2010年12月至2013年12月收治的HBV-ACLF患者共33例(HBVACLF组),选取同期住院的慢性乙型肝炎(CHB)患者42例(CHB组)和20例健康体检者(健康对照组)。采用ELISA法检测3组病例血清中IL-17与IL-35水平,同时检测各项肝功能指标。结果 HBV-ACLF组与CHB组患者IL-17、IL-35水平分别为(137.47±60.81)、(242.39±81.80)、(121.63±57.25)、(212.05±123.98)pg/mL,均高于健康对照组(16.37±8.47)、(33.6±24.63)pg/mL,(t=6.496、6.136、5.857、5.441,P0.01);HBV-ACLF组与CHB组IL-17、IL-35水平差异无统计学意义(t=0.987、1.034,P0.05)。结论 IL-17、IL-35的高水平表达可能与乙型肝炎慢加急性肝功能衰竭的发生有关。     

Immunoregulatory lymphokines in rheumatoid joints

Summary Since the role of interleukin-2 (IL-2) in rheumatoid synovial joints has been debated, we examined IL-2 production by, and IL-2 responsiveness of, cells eluted from synovial tissue (ST) of patients with rheumatoid arthritis (RA) and juvenile rheumatoid arthritis (JRA). IL-2 was not detected in unstimulated cell-culture supernatants from any of the four RA patients tested, but it was present in small amounts in supernatants of unstimulated cultures derived from three of seven JRA patients studied. After PHA stimulation, IL-2 was detected in corresponding supernatants from all RA and JRA patients and from normal mononuclear cells (MNC). There was no significant difference in IL-2 activity between supernatants of normal MNC and supernatants from either RA or JRA patients. The eluted cells showed a proliferative response to recombinant IL-2. Rheumatoid ST cells are thus able to produce and respond to IL-2. Since non-T cells present in the eluates might interfere with IL-2 metabolism, one cannot yet say whether T cells of rheumatoid ST themselves produce and respond to IL-2 in a normal or abnormal way.