类风湿性关节炎患者滑膜液中IL-10、IL-12和sFasL含量的检测

为了研究细胞因子和凋亡分子在类风湿性关节炎(RA)发病中的作用,用ELISA法分析了26例RA患者滑膜液和血清中IL-12、IL-10和可溶性FasL(sFasL)的含量。结果表明RA患者滑膜液中IL-12、IL-10含量分别为(419.9±89.2)pg/ml和(187.7±34.5)pg/ml,外周血中这两种细胞因子的含量均较低,分别为(65.3±34.2)pg/ml(IL-12)和(85.0±12.7)pg/ml(IL-10)。滑膜液中sFasL的含量为266pg/ml,明显高于血清含量(36pg/ml)。这一结果提示,RA患者滑膜液中IL-12含量和sFasL增高,这些炎性细胞因子增高可能参与了关节滑膜中的自身反应性T细胞的活化,继而造成免疫损伤。     

抑郁症患者血清细胞因子、C-反应蛋白和锌水平的变化及其临床意义

目的:探讨IL-6、IL-1β、TNF-α、CRP、Zn在抑郁症病理生理机制中的作用。方法:抑郁症患者33例,正常对照组23例。采用酶联免疫吸附法(ELISA)测定抑郁症患者和正常对照组的血清IL-6、IL-1β、TNF-α的水平,同时采用散射比浊法测定血清CRP,化学比色法测定血清Zn水平。于治疗前评定汉密尔顿抑郁量表(HAMD)。结果:①抑郁症血清IL-6[(8.90±5.63)pg/ml]、TNF-α[(13.57±7.63)pg/ml]、CRP[(6.18±5.68)mg/L]水平显著高于正常对照组(IL-6:5.95±3.66;TNF-α:12.87±5.34;CRP:2.50±1.44),且血清Zn水平[(11.88±2.37)μmol/L]显著低于正常对照组(13.60±1.90);抑郁症组血清Zn水平女性患者(11.19±2.21)显著低于男性患者(14.04±1.48)。②抑郁症血清IL-6水平与TNF-α有显著的正相关(r=0.470,P<0.01),而血清IL-1β水平与Zn有显著的正相关(r=0.346,P<0.05)。③抑郁症血清Zn与HAMD中迟缓因子分有显著的正相关(r=0.351,P<0.05),与性别有明显的正相关(r=0.576,P<0.01)。结论:①在抑郁症组血清IL-6、TNF-α、CRP水平升高可能是抑郁症的免疫学标志之一;②抑郁症中细胞因子IL-1β表达异常与微量元素锌密切相关,提示可能与抑郁症发病机制有关。     

血管内皮生长因子的检测在冠心病患者中的意义

目的:为了探讨血管内皮生长因子(VEGF)的检测在冠心病患者中的意义。方法:应用双抗夹心ELISA法检测了55例冠心病(CHD)患者和34例正常对照者的血清VEGF水平。结果:CHD患者血清VEGF水平(498.98±304.50)pg/ml明显高于正常对照组(121.49±30.29)pg/ml,P<0.01;其中急性心肌梗塞(AMI)患者组血清VEGF水平(809.21±191.16)pg/ml明显高于心绞痛患者组(292.14±148.56)pg/ml,P<0.01,而且两者均高于正常对照组。结论:提示VEGF可能是反映心肌缺血的敏感指标。     

动态观察高血压患者血清白细胞介素18改变的临床意义

探讨原发性高血压患者治疗前后血清白细胞介素18(IL-18)改变的临床意义. 采用酶联免疫分析法(ELISA)动态检测高血压患者治疗前后及对照组的血清IL-18含量. 高血压患者血清IL-8含量(1032.0±748.0pg/mL)明显高于对照组(420.6±385.3pg/mL), P＜0.05.1级高血压患者治疗前血清IL-8含量为955.2±808.0pg/mL, 治疗2周后血清IL-8含量为691.67±589.84pg/mL, 前后对照有明显差异(P＜0.01);2、3级高血压患者治疗前血清IL-18含量为1284.5±462.2pg/mL, 治疗2周后血清IL-18含量为881.84±474.04pg/mL, 前后对照有明显差异(P＜0.05).高血压患者血清IL-18含量明显高于对照组, 高血压患者治疗后血清IL-18含量明显低于治疗前血清IL-18含量, 因此提示 血清IL-18含量的改变可能参与高血压病的发病机制, 高血压患者存在免疫调节功能紊乱.     

白细胞介素-35对慢性乙型肝炎患者外周血Th17细胞的影响及其临床意义

目的观察白细胞介素(IL)-35在慢性乙肝患者外周血的水平和对Th17细胞的影响,为阐释慢性HBV感染的免疫耐受和炎症应答机制提供实验依据。方法 27例慢性乙肝患者和18例健康志愿者入组本研究。利用酶联免疫吸附试验检测外周血IL-35的水平,利用流式细胞术检测Th17细胞比例。应用重组人IL-35刺激外周血单个核细胞后观察Th17细胞和Th17相关细胞因子水平的变化,并观察抗病毒治疗后IL-35和Th17细胞水平变化。结果慢性乙肝患者血清IL-35水平(72.95pg/ml±15.71 pg/ml)较健康志愿者(46.35 pg/ml±13.71 pg/ml)显著升高(P0.000 1),且与HBV DNA呈显著正相关(P0.000 1)。慢性乙肝患者外周血HBV表位肽诱导Th17细胞比例(1.52%±0.46%)较健康志愿者(0.03%±0.01%)显著升高(P0.000 1),但非特异性Th17细胞在2组之间无显著差异(1.66%±0.42%vs 1.43%±0.43%,P=0.089)。在接受抗病毒治疗前,重组人IL-35刺激可显著降低慢性乙肝患者HBV表位肽诱导Th17细胞的比例和Th17细胞分泌的细胞因子(IL-17和IL-22)的水平。而抗病毒治疗可显著降低慢性乙肝患者血清IL-35水平,但对HBV表位肽诱导Th17细胞的比例无显著影响,同时,在接受抗病毒治疗后,重组人IL-35刺激对慢性乙肝患者Th17细胞和Th17细胞分泌细胞因子无明显影响。结论 IL-35在慢性乙型肝炎发病过程中可能发挥免疫抑制作用,导致机体免疫耐受和病毒持续感染。     

慢性乙型肝炎患者外周血单核细胞与血清HBV DNA定量对比研究

目的探讨慢性乙肝患者外周血单个核细胞(PBMC)内 HBV-DNA的出现与血清HBV-DNA浓度之间关系.方法应用荧光定量PCR技术检测57例慢性乙肝患者血清和PBMC中HBV-DNA含量、对血清中不同的病毒浓度进行分组比较分析.结果①57例PBMC内HBV-DNA总检测出率为42.1%(24/57),两者检测结果一致占88.6%.②根据血清内HBV-DNA浓度分成三组,三组PBMC内HBV-DNA浓度及阳性率比较p＜0.01,存在显著差异;③血清HBV-DNA阴性而 PBMC HBV-DNA阳性只有1例(1.7%),其浓度为1.50×108/L.结论①血清HBV-DNA浓度与PBMC内HBV-DNA的出现及HBV-DNA浓度存在明显相关性.②临床上检测PMBC中HBV-DNA是对血清HBV-DNA的一个重要补充;③PBMC内HBV-DNA检测对观察病毒在非血清内的状态、间接反映肝细胞病毒复制情况以及进一步指导抗病毒治疗有一定意义.     

IL—12以及IL—12P40对子宫内异症患者NK细胞功能的免疫调节

目的通过观察正常人及子宫内膜异位症(EM)患者外周血及腹腔液中IL-12、IL-12P40含量变化,分析其分子诱导NK细胞对子宫内膜细胞的细胞毒效应的影响,探讨IL-12、IL-12P40分子与EM患者NK细胞功能低下的相关性.方法ELISA酶联免疫检测IL-12、IL-12P40含量以及MTT释放法检测NK细胞毒活性.结果①EM患者腹腔上清液可使正常人外周血NK细胞杀伤活性(16.80%±3.6%)明显下降(8.37%±4.5% )(P＜0.05).② EM患者IL-12含量血清为66.38±12.6 pg/ml,低于对照组84.97±13.7 pg/ml(P＜0.05);腹腔液中为77.76±14.6 pg/ml,低于对照组106.92±10.7 pg/ml(P＜0.05).③EM患者IL-12P40含量血清为35.64±10.6 pg/ml, 与对照组无明显差异;腹腔液含量为79.76±12.6 pg/ml, 高于对照组40.54±10.0 pg/ml(P＜0.05), 并与疾病的程度呈负相关.④IL-12能增强NK细胞对子宫内膜细胞的杀伤,并呈剂量依赖.在10 ng/ml浓度诱导后杀伤率为31.90%,高于对照组的16.80%.⑤IL-12P40能拮抗IL-12对NK细胞活性的诱导,当加入IL-12P40后,使IL-12诱导的活性(29.3%)下降为19.36%.结论子宫内膜异位症患者腹腔液中IL-12P40含量的增高可能与NK细胞功能下降有关.     

肾综合征出血热患者血浆和血清可溶性IL-2R水平变化的研究

本文应用双McAb ELISA夹心法检测了HFRS患者血清和血浆中SIL-2R水平。结果表明,急性期患者血浆和血清中sIL-2R水平(分别为199±81U/ml和214±122U/ml)显著地高于恢复期患者(分别为104±37U/ml和97±28U/ml)和正常人(分别为88±31U/ml和71±21U/ml)(P<0.01);在急性期中尤以少尿期升高最明显。表明HFRS患者机体免疫细胞处于高度活化的状态,患者血清和血浆中sIL-2R的检测对于判断病期和指导治疗均有一定的意义。     

The therapeutic effect of Lamivudine on HBV-DNA in PBMC and its inducement effect against cytokine

AThe purpose of this investigation was to study the therapeutic effect of Lamivudine on HBV DNA in peripheral blood mononuclear cells (PBMC) and serum, and the level of cytokines in serum of the patients with chronic hepatitis B. The patients were divided into two groups (A = 47, B = 34), and treated by Lamivudine, routine medicine, respectively. The levels of HBV-DNA in PBMC and serum and cytokines were all detected before and after treatment. After the treatment of Lamivndine for 36 weeks, the total conversion negative rates of HBV-DNA in PBMC and serum of the patients treated with Lamivudine were 55.32% (26/47) and 61.70% (29/47), respectively. The total negative conversion rates of HBV-DNA in PBMC and serum of the patients treated by routine medicine were 26.47% (9/34) and 32.35% (11/34), respectively. There was significant difference between Lamivudine group and routine medicine group ( P < 0.01). The negative conversion rates of HBeAg in serum of the patients were 46.81% (22/47) and 68.09% (32/47) at the end of 24 weeks and 36 weeks, and were higher than those of routine medicine group ( P < 0.05 and P < 0. 01). The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), ALT/AST in serum of the patients after being treated by Lamivudine, routine medicine were down-regulated to (30.1±9.6) U/ml, (32.3±10.7) U/ml, 0.9±0.1 and (48.4±10.7) U/ml, (44.7±11.0) U/ml, 1.1±0.2. After the analysis of variance, the high significant difference was obvious between the two groups (P < 0.01). It was due to the high levels of IL-6, IL-8 and TNF-αin chronic hepatitis B which could be down-regulated to (250. 5±33. 3) pg/ml, (153.4±22.2) pg/ml, (232.6±21.2) pg/ml by Lamivudine, which was more obvious than that of routine medicine (P < 0.01). Lamivudine has high therapeutic effect on the treatment of HBV DNA in PBMC and serum and has better therapeutic effect than that of routine therapy. Lamivudine may also have higher down-regulated inflammatory infiltration and secretion in local site caused by chemotactic cytokines and produce promotional effect on the recovery of liver function.     

白三烯D4和白介素13对人肺成纤维细胞白三烯受体1 mRNA表达及嗜酸粒细胞趋化因子产生的影响

探讨白三烯D4(LTD4)和白介素13(IL-13)对人肺成纤维细胞白三烯受体(CysLT1R)表达和eotaxin产生的影响。采用培养人肺成纤维细胞株(MRC-5),以不同浓度的IL-13进行刺激,用实时定量RT-PCR法测定CysLT1R mRNA表达的变化。再以不同浓度的LTD4和IL-13刺激,用ELISA法测定上清液中eotaxin的浓度,并观察Montelukast对eotaxin的产生是否有抑制作用。结果显示,培养的人肺成纤维细胞上存在着CysLT1R mRNA的低表达,以10 ng/ml IL-13刺激时,随着刺激时间延长,CysLT1R mRNA表达逐渐增高,48 h达高峰,较未刺激时表达增加(18.56±7.41)倍(P<0.01)。当以10 ng/ml和100 ng/ml的IL-13浓度刺激48 h时,较未刺激细胞CysLT1R mRNA表达分别增加(17.33±3.81)倍和(20.11±5.05)倍(P<0.01)。IL-13也能促进成纤维细胞产生eotaxin,以10 ng/ml和100 ng/ml的IL-13刺激成纤维细胞时,eotaxin浓度分别为(155.43±15.95)pg/ml和(221.31±17.23)pg/ml,较未刺激细胞(31.67±3.49)pg/ml明显增加(P<0.01)。单独给予LTD4刺激时,eotaxin浓度则无明显变化。但是以1×10-7mol/L和1×10-6mol/L浓度的LTD4分别联合10 ng/ml的IL-13共同刺激时,产生eotaxin的浓度分别为(204.4±25.4)pg/m和(255.1±38.3)pg/ml;较仅给予10 ng/ml的IL-13刺激时(155.4±16.0)pg/ml明显增加(P<0.01)。在LTD4+IL-13刺激组中,加入Mointelukast后eotaxin浓度为(148.3±15.7)pg/ml,较不加Montelukast(204.4±25.4)pg/ml明显降低(P<0.01)。人肺纤维细胞存在着CysLT1R mRNA的低表达,IL-13能够上调其表达,呈现时间与浓度依赖性。IL-13和LTD4对人肺成纤维细胞分泌的eoitaxin具有协同刺激作用,这可能与IL-13上调成纤维细胞CysLT1R mRNA表达有关,而Montelukast对这种刺激具有拮抗作用。     

Cryptic association of e antigen with different morphologic forms of hepatitis B surface antigen

When highly purified HBsAg particles, separated by rate zonal centrifugation into populations differing in predominant size, were tested for HBeAg, the e1 specificity was detected preferentially in association with particle fractions containing large filaments and Dane particles. These results were obtained both by agar gel diffusion and by radioimmunoassay for e antigen. The e antigen activity present in these fractions was potentiated by prior treatment of particles with Tween 80, suggesting cryptic localization of e1 specificity within or under the outer membrane. The HBeAg released by detergent treatment from a purified preparation composed predominantly of small-particle forms of HBsAg was separated by electrofocusing into a peak of nonparticulate e antigen in the pH range of 5.7--6.0. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed three major polypeptides in this preparation with approximate molecular weights of 25,000, 55,000, and 70,000. Furthermore, two additional peaks of e antigen activity were detected which migrated in association with HBsAg particles at isoelectric points of 4.4 and 5.5--5.6. The major portion of e antigen remained in association with particles after further purification by rate zonal centrifugation.     

HBIG联合HBVac阻断HBV宫内感染的研究

目的探讨孕期多次联合注射乙型肝炎免疫球蛋白 (HBIG)和乙型肝炎疫苗(HBVac )阻断乙型肝炎病毒HBV)宫内感染效果.方法对30例HBsAg阳性孕妇从孕20周开始联合注射HBIG和HBVac,另23 例未行注射的HBsAg阳性孕妇作对照.采用敏感特异的套式聚合酶链反应检测孕妇及新生儿血清及外周血单个核细胞(PBMC)中HBV DNA.结果阻断组和对照组新生儿血清HBV DNA检出率分别为10%(3/30)和34.8%(8/23),二组差异显著,而PBMC中HBV DNA检出率二组无显著性差异;阻断组孕妇血清 HBV DNA水平下降率38.5 %(10/26),而对照组10.5%(2/19),差异显著, PBMC中HBV DNA水平二组无显著性差异.结论孕期多次联合注射HBIG 和 HBVac 可有效降低孕妇体内HBV DNA水平,从而降低HBV宫内感染率,但对孕妇PBMC中 HBV 影响不大, 并不降低新生儿 PBMC 中HBV DNA的检出率.     

Secular trend of age-specific prevalence of hepatitis B surface and e antigenemia in pregnant women in Taiwan

To elucidate the impact of aging of hepatitis B carrier women on their viral replicative markers in a hepatitis B endemic area, all the parturients admitted to the Hospital were studied from 1985 to 2000. Serum hepatitis B surface (HBsAg) and hepatitis B e antigen (HBeAg) were tested by radioimmunoassay. Mann-Whitney U and Student's t-tests were used for statistical analysis. The results showed the yearly prevalence rate of HBsAg in pregnant women seemed stable with a mean of 12.0 +/- 1.1% during the period. The yearly positive rate of HBeAg among HBsAg-positive pregnant women varied between 30.4% and 42.6% from 1985 to 1992 and declined from 29.6% in 1993 to 18.1% in 2000. The mean ratio of HBeAg/HBsAg in carrier parturients was 24.7% [intraquantile range (IQR) 20.5-28.4] from 1993 to 2000, which was significantly lower than that of 32.4% (IQR 31.0-39.0) from 1985 to 1992 (P < 0.0001). The mean age of HBeAg-positive primiparas from 1993 to 2000 was 29.1 +/- 3.9 years and significantly higher than that of 28.0 +/- 3.7 years from 1985 to 1993 (P < 0.001), as well as in secundiparas 31.2 +/- 3.8 years vs. 30.1 +/- 3.4 years (P < 0.001) and in total parturients 30.3 +/- 4.2 years vs. 29.3 +/- 3.8 years (P < 0.001). Thus, no significant decrease of HBsAg carriage was observed in the past 16 years, whereas a decreased ratio of HBeAg/HBsAg was noted in carrier parturients in the past 8 years and the elderly HBeAg-positive parturients from 1993 to 2000 may be the cause.     

Prevention of perinatal transmission of hepatitis B virus (HBV): a comparison of two prophylactic schedules

Perinatal transmission of hepatitis B virus (HBV) from HBsAg carrier mothers who were HBeAg+, antiHBe+, or negative for both HBe markers, was interrupted using either 4 doses of vaccine, or one dose of hepatitis B immunoglobulin (HBIG) at birth, combined with 4 doses of vaccine. In those infants who received HBIG at birth, the antiHBs titre was significantly higher at 1 and 2 months old, but at 6, 9, and 18 months old, there was no significant difference. Among the infants of carrier mothers who did not display HBeAg (i.e., were antiHBe+, or negative for both HBe markers), a transient subclinical infection would have been expected in around 10% had there been no intervention. No evidence of such infection was detected, and no difference in outcome was found between the two treatment groups. Amongst infants born to HBeAg+ carrier mothers, infection occurred in 1 out of 8 who had received HBIG and vaccine, and in 3 of 8 who had received vaccine only. The difference in outcome was not statistically significant, but the numbers analysed were small. The infections which occurred in spite of prophylaxis may be attributable to in utero infection, poor response to vaccine by the infant, or to the mother having a particularly high HBV-DNA level. HBIG given at birth to infants of HBeAg+ carrier mothers may enhance the protection of infants who are destined to be poor responders to vaccine.     

慢性乙肝患者树突状细胞抗原递呈功能状态的初步研究

目的:研究慢性乙肝患者树突状细胞(DC)抗原递呈功能的改变。方法:从慢乙肝患者和健康人外周血分离单个核细胞,诱导培养出DC,显微镜下观察DC形态并计数,流式细胞仪检测DC表面协同刺激分子(CD80,CD86),MTT法检测DC刺激同种异体淋巴细胞增殖的能力,ELISA法测定培养上清液中IL-12、r-IFN水平。结果:慢乙肝患者外周血单个核细胞诱导培养所获得的DC数量及表面协同刺激分子(CD80,CD86)表达水平、刺激同种异体淋巴细胞增殖能力和细胞因子产生水平均明显低于健康者(P<0.05)。结论:慢乙肝患者DC的数量及抗原递呈、免疫刺激功能状态均处于低下水平。     

Relationship between serum hepatitis B virus DNA and surface antigen with covalently closed circular DNA in HBeAg‐negative patients

Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) is responsible for viral persistence. This study aimed to investigate the serum surrogate markers for cccDNA and to evaluate the intrahepatic viral events associated with disease activity in HBeAg‐negative chronic hepatitis B patients. Thirty‐three treatment‐naïve patients with a negative HBeAg who had a liver biopsy were studied. Active disease was defined as a serum alanine aminotransferase >40 IU/L and a serum HBV DNA >10,000 copies/ml. This study showed significant correlation between serum HBV DNA and both log cccDNA (r = 0.41, P = 0.018) and log total intrahepatic HBV DNA (r = 0.71, P < 0.0001). No significant correlation was observed between serum HBsAg and log cccDNA (P = 0.15) or log total intrahepatic HBV DNA (P = 0.97). Fourteen and 19 patients had inactive and active disease, respectively. The median log cccDNA and log total intrahepatic HBV DNA (copies/10⁶ cells) were significantly higher in patients with active disease compared with those with inactive disease (4.11 vs. 3.53, P = 0.03 and 5.46 vs. 4.64, P < 0.001, respectively). The HBV replicative efficiency, defined as the ratio of serum HBV DNA to cccDNA, was approximately 20% higher in patients with active disease. No significant difference was observed in the HBsAg levels and the ratio of serum HBsAg to cccDNA between the two groups. In conclusion, serum HBV DNA, but not HBsAg, reflects the amount of cccDNA and the replication efficiency of HBV in patients with HBeAg‐negative chronic hepatitis B. J. Med. Virol. 82:1494–1500, 2010. © 2010 Wiley‐Liss, Inc.     

Gender disparity in distribution of the major hydrophilic region variants of hepatitis B virus genotype C according to hepatitis B e antigen serostatus

Hepatitis B virus (HBV) e antigen (HBeAg) seroconversion during chronic HBV infection is known to play an important role in disease progression and patient response to antiviral agents. The aim of the present study was to analyze gender disparity in distribution of major hydrophilic region (MHR) variants according to HBeAg serostatus. Prevalence of MHR variants from 68 Korean patients with chronic hepatitis (31 HBeAg-positive and 37 HBeAg-negative) was examined in terms of HBeAg serostatus and sex by direct sequencing analysis of the MHR. Gender disparity was observed in the distribution of MHR variants according to HBeAg serostatus. In male patients, the prevalence of MHR variants was significantly higher in HBeAg negative patients than in HBeAg positive patients [58.8% (10/17 patients) vs. 14.3% (3/21 patients), P=0.004]. However, the same was not true in female patients [55.0% (11/20 patients) vs. 60.0% (6/10 patients), P=1.000)]. In addition, 2 mutation types (L110I and G145A) related to HBeAg serostatus were found. In conclusion, HBeAg seroconversion in male chronic patients infected with genotype C could lead to mutations of MHR, major target to host immune response, which might in turn contribute to HBV persistence and immune evasion.     

B7家庭新成员

T细胞活化除需要TCR传导的第1信号外,还需协同刺激分子传导的第2信号.