外周血单核细胞SOCS-1表达与MODS患者预后关系的研究

目的了解外周血单核细胞SOCS-1表达与多器官功能障碍综合征（MODS）患者预后的关系及临床意义。方法收集24例MODS患者，并采集其外周静脉血，采用淋巴细胞分离液密度梯度离心法分离外周血单核细胞（PBMCs），分别以RT-PCR法及Western-blot法检测PBMCs中SOCS-1的基因及蛋白表达，分析其与预后及MODS评分的关系。结果MODS组中死亡患者PBMCs中的SOCS-1mRNA表达量（0.4938±0.0273）显著低于存活患者（0.5475±0.0289）（P〈0.05），SOCS-1蛋白表达量（0.7924±0.0284）显著低于存活患者（0.8406±0.0407）（P〈0.05）。MODS患者的PBMCs中的SOCS-1mRNA表达量与MODS评分呈显著的负相关关系（r=-0.723,P〈0.01），SOCS-1蛋白表达量与MODS评分呈显著的负相关关系（r=-0.534，P〈0.01）。结论在MODS中，SOCS-1的表达可能起到保护组织避免损伤的作用,SOCS-1表达的减少可能提示患者的预后不良     

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 5 7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　① 5 7例PBMC内HBV DNA总检测出率为 4 2 .1% (2 4 / 5 7) ,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 .0 1,存在显著差异 ;③血清HBV -DNA阴性而PBMCHBV -DNA阳性只有 1例 (1.7% ) ,其浓度为 1.5 0× 10 8/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     

狼疮性肾炎患者外周血IL-18水平及其基因表达

为了探讨白细胞介素 18(IL 18)在狼疮性肾炎 (LN )发生、发展中的作用。我们采用逆转录多聚酶链反应 (RT PCR )及酶联免疫吸附 (ELISA )法测定 16例正常人及 18例LN患者外周血单个核细胞 (PBMC )IL 18mRNA表达量及其血浆水平。结果提示LN患者PBMCIL 18mRNA表达量及血浆IL 18水平均较正常对照组显著增高 [IL 18mRNA表达量为 :1 2 6 2± 0 189vs0 84 4± 0 15 5 ,P <0 0 0 1;IL 18血浆水平为 :(82 2 0 9± 5 32 77)pg/mlvs (2 39 5 7± 75 0 6 )pg/ml,P <0 0 0 1]。且WHOIV型LN增高较非IV型LN更为显著 [IL 18mRNA表达量为 :1 32 9± 0 2 1vs 1 138± 0 15 2 3,P <0 0 5 ;IL 18血浆水平为 :(1135 5 4± 5 15 34)pg/mlvs (5 0 8 6 5± 341 36 )pg/ml,P <0 0 1]。另外 ,血浆IL 18水平与肾组织活动指数 ,肾小管间质损害程度呈等级相关 (r分别为 :0 6 10和 0 4 99,P均 <0 0 5 ) ,也与血清肌酐 (Scr) ,血清内生肌酐清除率 (Ccr)及 2 4h尿蛋白排泄量 (2 4hUPQ )呈直线相关 (r分别为 :0 898、 0 6 2 8和 0 5 37,P均 <0 0 5 )。本研究认为循环IL 18表达和分泌增高可能参与LN的免疫发病过程 ,并与狼疮活动有一定的关系     

宫颈癌患者外周血CD2^＋细胞表达的检测及意义

目的：探讨宫颈癌患者外周血CD2^＋细胞表达水平及临床意义。方法：应用流式细胞术检测40例宫颈癌患者及30例宫颈上皮内瘤变（cervical intra-epithelial neoplasia,CIN）患者外周血CD2^＋细胞百分率,以30例子宫肌瘤患者作为正常对照。结果：宫颈癌患者外周血CD2^＋细胞百分率水平明显低于CIN组及正常对照组,差异均具有统计学意义（P〈0.05）。结论：CD2^＋的表达与宫颈癌的发生、发展有关,可作为宫颈癌患者治疗及评价预后的参考指标。     

Expression and Functional Analysis of Toll-Like Receptors of Peripheral Blood Cells in Asthmatic Patients: Implication for Immunopathological Mechanism in Asthma

Background  We investigated the expression profile of toll-like receptors (TLRs) and TLR ligand-activated production profile of asthma-related inflammatory cytokines in asthmatic patients. The expression of TLR1–8 on monocytes, CD4+ T helper lymphocytes, CD8+ T cytotoxic lymphocytes, CD19+ B lymphocytes, and dendritic cells, and ex vivo production of cytokines from peripheral blood mononuclear cells activated by TLR ligands were measured by flow cytometry. Discussion  Ex vivo productions of TNF-α, IL-10, and IL-1β by TLR4 and TLR5 ligand LPS and flagellin were significantly lower in asthmatic patients (all P < 0.05). Expression of TLR4 and TLR5 was also found to be significantly lower in asthmatic patients when compared to that of control subjects (all P < 0.05). Therefore, the decreased activation of TLR4 and TLR5 in asthmatic patients might contribute to the immunopathological mechanisms of asthma by reducing the release of Th1 and anti-inflammatory cytokines. Samantha W. M. Lun and C. K. Wong contributed equally to this study.     

慢性乙型肝炎患者外周血CD4＋CD25＋、CD8＋CD28＋淋巴细胞的变化及与病毒载量的关系

目的：研究慢性乙型肝炎患者外周血CD4＋CD25＋、CD8＋CD28＋淋巴细胞的表达变化及与病毒载量的关系。方法：利用流式细胞术检测50例慢性乙型肝炎患者外周血CD4＋CD25＋、CD8＋CD28＋淋巴细胞的表达率和绝对数,并与正常对照组30例进行比较;荧光定量PCR检测慢性乙型肝炎患者HBV核酸的载量,并与CD4＋CD25＋、CD8＋CD28＋淋巴细胞的表达进行相关分析。结果：慢性乙型肝炎患者的CD4＋CD25＋表达率和细胞数分别为（15.60±5.86）%和（0.34±0.13）×109/L,明显高于正常对照组（P〈0.01）;而CD4＋CD25-、CD8＋CD28＋为（21.13±5.32）%、（0.47±0.19）×109/L和（9.49±2.57）%、（0.21±0.07）×109/L,均低于正常对照组（均P〈0.01）;不同病毒载量的慢性乙型肝炎患者CD4＋CD25＋T细胞与病毒载量均呈正相关（r分别为0.552、0.588,P均〈0.01）,而CD8＋CD28＋T细胞与病毒载量均无相关性（r分别0.275、-0.092,P均〉0.05）。结论：慢性乙型肝炎患者细胞毒T细胞（CD8＋CD28＋）减少,免疫调节细胞的增加与慢性乙型肝炎患者病毒长期存在,病程迁延不愈有关。     

川芎嗪和地塞米松对狼疮肾炎患者PBMC IL-12表达及合成Ig的影响

应用细胞培养技术 ,采用半定量RT PCR法和ELISA检测等方法 ,以正常人为对照 ,观察川芎嗪 (33μg/ml)和地塞米松 (10 5mol/L )对培养的狼疮肾炎 (LN )活动期、静止期外周血单个核细胞 (PBMC )表达IL 12和产生免疫球蛋白 (Ig)的影响。结果表明 ,LN活动期、静止期较正常对照组IL 12蛋白、基因水平明显增高 (P <0 0 1) ,川芎嗪仅对活动期狼疮肾炎PBMCIL 12表达明显抑制 ,而地塞米松明显抑制狼疮肾炎PBMCIL 12表达 (P <0 0 1) ,两种药物对正常人对照IL 12表达及合成Ig无明显抑制作用 (P >0 0 5 ) ,川芎嗪和地塞米松均明显抑制狼疮肾炎活动期及静止期Ig合成 (P <0 0 1或 <0 0 5 )。提示 :狼疮肾炎患者有高水平的IL 12 ,川芎嗪、地塞米松通过下调狼疮肾炎IL 12表达及Ig的合成 ,减轻狼疮肾炎的发生和发展。     

血清CD28、CTLA4及LAG3在慢性重型乙型肝炎患者的表达水平及其对短期预后的预测价值

目的探究慢性重型乙型肝炎患者血清白细胞分化抗原28(CD28)、细胞毒性T淋巴细胞相关抗原(CTLA4)及淋巴细胞活化基因3蛋白(LAG3)水平及其对患者短期预后的预测价值。方法选择2016年2月至2019年4月我院收治的107例慢性重型乙型肝炎、59例慢性乙型肝炎患者、50例健康体检者作为研究对象,分别纳入慢重肝组、慢乙肝组及健康组。收集各组受试者临床资料,测定血清CD28、CTLA4、LAG3水平,随访慢重肝组患者预后情况。结果①与健康组比较,慢重肝组、慢乙肝组患者天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、总胆红素(TBIL)、HBV-DNA载量及血清CTLA4、LAG3等指标水平升高(P<0.05),白蛋白(ALB)、CD28水平降低(P<0.05),慢重肝组各指标升高或降低幅度大于慢乙肝组(P<0.05)。②HBV-DNA载量与血清CD28水平呈负相关(P<0.05),与血清CTLA4、LAG3水平呈正相关(P<0.05)。③慢重肝组患者3个月内死亡率为48.60%,死亡组患者血清CD28低于存活组,CTLA4及LAG3水平显著高于存活组(P<0.05)。④血清CD28、CTLA4及LAG3预测慢重肝患者死亡的曲线下面积分别为0.777、0.792、0.774。⑤CD28高表达组患者生存率显著高于CD28低表达组(P<0.05);CTLA4高表达组、LAG3高表达组生存率显著低于CTLA4低表达组、LAG3低表达组(P<0.05)。结论血清CD28、CTLA4及LAG3与慢性重型乙型肝炎患者病情及预后有一定相关性,临床可考虑将其用于病情严重程度、预后评估的辅助指标。     

肺癌患者CD4+ CD25+调节性T细胞的检测及临床意义

目的:检测肺癌患者外周血CD4 CD25 调节性T细胞的分布并探讨相关机制.方法:采用流式细胞仪分析66例肺癌患者外周血CD4 CD25 调节性T细胞占CD4 T淋巴细胞的比例.结果:66例肺癌患者外周血中CD4 CD25 调节性T细胞占CD4 T淋巴细胞的比例为(16.2±2.4)%,与对照组(6.19±1.5)%比较差异有显著性(P＜0.05). 25例鳞癌、29例腺癌、12例小细胞癌患者外周血中CD4 CD25 调节性T细胞比例分别为(18.3±2.9)%、(15.6±1.8)%、(17.3±2.2)%,各组间比较差异无显著性(P＞0.05);均显著高于对照组(6.19±1.5)%,P＜0.05;34例Ⅲ期,14例Ⅳ期肺癌患者外周血中CD4 CD25 调节性T细胞比例为(15.3±2.6)%,(20.4±3.1%),均显著高于18例Ⅱ期患者(9.4±1.3)%,P均＜0.05.结论:肺癌患者外周血中有CD4 CD25 调节性T细胞比例增高,且与分期有关.它可能与肺癌患者免疫功能受损有关,可作为评估肺癌患者预后的一项指标.     

新城鸡瘟病毒诱导人粘附性单个核细胞产生一氧化氮与细胞毒性

本文采用Griess试剂、间接免疫荧光法和同位素释放等方法,发现新城鸡瘟病毒Losota系（NDV-L）与人外周血粘附性单个核细胞（a-MCs）作用2～4h后,可稳定地吸附在a-PBMCs表面,并使其释放一氧化氮（NO）,释放量与阳性对照组（BCG-LPS作用的a-PBMCs）相近;采用~3H-TdR释放法测定NDV-L作用的a-PBMCs对K_(562)靶细胞的细胞毒活性,发现具有明显的杀伤效应,且该杀伤效应与NO的产生有一定的依赖性。     

Increased bcl-2 expression in lymphocytes and its association with hepatocellular damage in patients with autoimmune hepatitis

The proto-oncogene product bcl-2 is known to inhibit apoptotic cell death, and its dysregulation might play a critical role in the development of autoimmune disease. To elucidate the role of bcl-2 in autoimmune hepatitis (AIH), its expression in peripheral blood mononuclear cells (PBMC) and in liver-infiltrating lymphocytes (LIL) was investigated. Increased bcl-2 expression in PBMC was found in AIH patients compared with that in chronic hepatitis C (CHC) patients and in healthy controls. The level of bcl-2 expression significantly correlated with serum ALT level. Further analysis showed that CD4+ T cells are enriched in bcl-2-expressing PBMC. To characterize the Th1/Th2 profile of bcl-2-expressing CD4+ T cells, intracellular interferon-gamma (IFN-gamma) and IL-4 were analysed. The results revealed that most of the bcl-2-expressing cells were found to be IFN-gamma-secreting Th1 cells. In three patients for whom their clinical courses could be followed, bcl-2 expression was decreased after the initiation of immunosuppressive therapy with corticosteroids. However, the level of IFN-gamma + cells was not altered. Immunohistochemical analysis also showed that large amounts of bcl-2+ cells were observed in periportal area in the liver. In conclusion, bcl-2-expressing cells were shown to be increased in peripheral blood and liver in AIH and the bcl-2 product was expressed mainly in CD4+ Th1-type cells, suggesting that these cells might promote the cellular immune response and contribute to the development of hepatitis and hepatocellular damage in AIH.     

护肝合剂对慢性乙型肝炎患者PBMC功能的影响

目的观察护肝合剂对慢性乙型肝炎患者肝功能、外周血单个核细胞（PBMC）功能的影响。方法治疗组50例．在一般治疗的基础上加用护肝合剂，对照组15例用甘草酸二胺氯化钠溶液、门冬氨酸鸟氨酸、灯盏花素等进行一般治疗。分离培养治疗前后外周血单核细胞，加入植物血凝素（PHA），培养48h后，1500r／min离心10min，收集上清液-20℃保存，统一检测IFN-y，IL-10含量。采用流式细胞仪检测治疗前后外周血CD8^＋CD28^＋T细胞亚群。结果治疗组谷丙转氨酶（ALT）、谷草转氨酶（AST），黄疸指数（TBIL）复常率均明显优于对照组（P〈0．05，P〈0．01，P〈0．01）。治疗组治疗后IFN-y显著升高（P〈0．05），IL-10显著下降（P〈0．05），对照组升高不显著（P〉0．05）。CD8^＋CD28^＋T细胞亚群治疗组较对照组显著升高（P〈0．05），且治疗组治疗前后亦显著升高（P〈0．05）。结论护肝合剂能够改善肝功能，提高慢性乙型肝炎患者Th1细胞免疫。     

Characteristics of a Specific Radioimmunoassay for Measurement of Ferritin on the Surface of Peripheral Mononuclear White Blood Cells in Cancer Patients

Using ¹²⁵I-labeled rabbit anti-Hodgkin's spleen ferritin antibody (RHF), a simple radioimmunoassay has been developed for quantitation of ferritin on the surface of peripheral blood mononuclear white blood cells (PBM). This method makes use of a % specific binding determination (%SP) by measuring the amount of ¹²⁵I-labeled RHF bound to 1 × 10⁶ PBM in the presence and absence of soluble ferritin. To standardize this procedure, artificial ferritin positive control cells were prepared by covalently coupling ferritin to cultured acute lymphoblastic luekemia cells. These cells were tested on a daily basis in parallel with patient PBM's to ensure inter and intra-assay precision and remained stable for over two years. Characteristics of ¹²⁵I-labeled RHF binding to control and patient PBM's were evaluated to determine the specificity of interaction and optimum binding parameters. %SP was linear in the range of 1 × 10⁵ - 1 × 10⁶ PBM's and was progressively inhibited by graded concentrations of soluble ferritin. F(ab')₂ preparations of RHF were equally as effective as intact RHF in blocking ¹²⁵I-labeled RHF binding confirming that ¹²⁵I-labeled RHF was not binding non-specifically to PBM Fc receptors. Additional experiments describing kinetics and methods of standardization of new lots of ¹²⁵I-labeled RHF are also described.     

低剂量辐射对外周血单个核细胞体外抗肿瘤活性的影响

目的：观察低剂量γ射线辐照的外周血单个核细胞对体外培养人胃癌细胞系MKN-28细胞的杀伤作用的影响。方法：实验设MKN-28肿瘤细胞对照组、外周血单个核细胞对照组、照射和未照射的外周血单个核细胞与肿瘤细胞共培养组，照射组的照射剂量为1Gy。利用吖啶橙／溴化乙锭（AO／EB）荧光双染色法，定期观察外周血单个核细胞对肿瘤细胞的杀伤情况。结果：外周血单个核细胞照射后培养至144h时有大量细胞死亡，存活细胞明显少于未照射组，培养至240h时．照射与非照射组中存活的外周血单个核细胞均减少，但是在照射组中减少更明显一些，在外周血单个核细胞与MKN-28肿瘤细胞共培养组中，照射组在共培养96h-240h时，外周血单个核细胞对胃癌细胞的杀伤作用比未照射组明显增强。结论：γ射线辐照对某些外周血单个核细胞有损伤作用，低剂量辐射的外周血单个核细胞杀伤肿瘤细胞活性增强，但肿瘤细胞对辐照后的外周血单个核细胞有趋化及细胞毒作用。     

Characteristics of Expanded CD4+CD28null T Cells in Patients with Chronic Hepatitis B

Expanded CD4⁺CD28^null T cells have not been described in the circulation of patients with chronic hepatitis B (CHB). The aim of the present was to detect and characterize the surface phenotype and functional capacity of these cells in CHB patients. Expanded CD4⁺CD28^null T cells were detected in the circulation of CHB patients with high viral load and elevated aminotransferase levels. Most CD4⁺CD28^null T cells showed a CD27?CD45RA^?CD45RO⁺ surface phenotype. The markers CD56, CD57 and killer immunoglobulin-like receptor (KIR) were detected on CD4⁺CD28^null T cells, but the majority were positive for CD57. Functionally, CD4⁺CD28^null T cells were found to be potent cytotoxic T lymphocytes with perforin and granzyme B secretion profiles. These findings indicate that the expanded CD4⁺CD28^null T cells are cytotoxic memory T cells and display a distinct functional phenotype in comparison with CD4⁺CD28⁺ T cells. The presence of these cells appears to be associated with inflammatory conditions, suggesting that these elevated CD4⁺CD28^null T cells might be involved in the pathogenesis of CHB.     

RA患者外周血CD8＋CD28-和CD4＋CD25＋调节性T细胞的表达及疾病活动性指标相关性分析

目的：检测类风湿性关节炎（RA）患者外周血CD8＋CD28-、CD4＋CD25＋调节性T细胞亚群,探讨其与临床活动性指标的关系。方法：采用流式细胞术检测台州医院RA患者外周血CD8＋CD28-、CD4＋CD25＋ T细胞亚群比例,探讨调节性T细胞与RA活动性、类风湿因子（RF）、免疫球蛋白（Ig）、C反应蛋白（CRP）、补体C3、抗CCP抗体、抗核抗体（ANA）、血小板（PLT）及血沉（ESR）的关系。结果：活动期RA患者外周血CD4＋CD25＋调节性T细胞亚群比例显著低于正常对照组（P〈0.01）,但稳定期RA患者与正常对照组结果差异无统计学意义（P〉0.05）。活动期和稳定期RA患者CD8＋CD28-与正常对照组相比较,结果无统计学意义（P〉0.05）;CD4＋CD25＋与CRP密切相关（r=-0.593,P〈0.05）,CD8＋CD28-与ESR相关系数呈弱相关。CD4＋CD25＋和CD8＋CD28-细胞与RF、IGG、C3、ANA、anti-CCP和PLT未见明显相关性。结论：活动期RA患者外周血CD4＋CD25＋ T细胞亚群比例减少,CD4＋CD25＋ T细胞可能与类风湿性关节炎疾病进展有关。     

Nigerooligosaccharides Augments Mitogen-Induced Proliferation and Suppresses Activation-Induced Apoptosis of Human Peripheral Blood Mononuclear Cells

Nigerooligosaccharides (NOS), a mixture of nigerose and nigerosylmaltooligosaccharides, consists of immunopotentiating oligosaccharides found in foodstuffs. We have previously reported that activation of peripheral blood mononuclear cells (PBMC) in response to concanavalin A (Con A) or a streptococcal preparation of OK-432 is augmented in healthy young adults and elderly subjects after the intake of NOS-supplemented syrup. A reappraisal of the data suggests that NOS augments proliferation but partly suppresses activation-induced apoptosis of PBMC in response to these mitogens. To confirm this hypothesis, PBMC from healthy male subjects were stimulated with Con A or OK-432 in the presence of nigerose at the concentrations at which it was detected in the blood of subjects who had ingested NOS-supplemented syrup. Cellular activation, specifically metabolic demand, viability and proliferation, was assessed from glucose consumption, by WST-1 colorimetry and by 5-bromo-2'-deoxy-uridine incorporation assay, respectively. The Con A-induced activation of PBMC in each measurement was significantly augmented by nigerose. OK-432-induced decreases in the viability of PBMC were significantly inhibited by nigerose. Stimulation of PBMC with Con A or OK-432 induced apoptosis, but nigerose suppressed such activation-induced cell death. These results indicated that nigerose activated PBMC in vitro in a manner similar to the process observed in vivo, providing further evidence for the effectiveness of consumption of NOS-supplemented syrup.     

系统性红斑狼疮外周血单个核细胞CD28/B7分子mRNA的表达

目的 :探讨CD2 8 B7分子在系统性红斑狼疮 (SLE)发病机制中的作用及其临床意义。方法 :应用逆转录 聚合酶链反应 (RT PCR)检测 35例活动期SLE患者和 30例正常人外周血单个核细胞 (PBMC)中CD2 8、B7 1和B7 2mRNA的表达水平。结果 :35例活动期SLE患者PBMC中CD2 8的阳性表达率 (2 2 86 % )明显低于正常人对照组 (70 0 0 % ) ,差异非常显著 (P <0 0 0 1) ;B7 2的阳性表达率 (82 86 % )明显高于正常对照组 (5 3 33% ) ,差异显著 (P <0 0 1) ;活动期SLE组CD2 8的平均表达水平 (0 194 5± 0 2 0 74 )明显低于正常对照组 (0 4 2 38± 0 10 5 3) ,差异显著 (P <0 0 5 ) ;B7 2的平均表达水平 (0 86 75± 0 2 5 75 )明显高于正常人对照组 (0 4 898± 0 30 72 ) ,差异非常显著 (P <0 0 1) ;35例活动期SLE患者中仅有 2例B7 1呈阳性表达。结论 :CD2 8 B7分子的异常表达可能与SLE患者淋巴细胞和抗原呈递细胞 (APC)的功能变化有关。B7 1低水平与B7 2的高水平表达表明 ,SLE患者T细胞的活化可能主要是通过CD2 8与B7 2的交联传递共刺激信号 ,介导以Th2型反应为主的免疫应答反应 ;B7 2的表达水平可能与SLE疾病的活动性有一定的相关性。CD2 8mRNA的低水平表达可能与外周血CD2 8 T细胞凋亡增加或迁移到炎症部