妇科千金片对慢性盆腔炎模型大鼠子宫组织辅助性T细胞1/2型细胞因子表达的影响

目的研究妇科千金片对慢性盆腔炎大鼠组织辅助性T细胞（helper T cell,Th）1/2型细胞因子表达的影响。方法采用金黄色葡萄球菌、大肠杆菌及解脲脲原体混合菌接种法建立慢性盆腔炎模型。模型成功后,随机分成妇科千金片低、中、高（052,104,208 g/kg）剂量组和假手术组、模型组、空白组6组。治疗21 d后,取子宫组织,采用酶联免疫吸附测定法检测白介素 1β（interleukin 1β,IL 1β）、IL 8、肿瘤坏死因子 α（tumor necrosis factor α,TNF α）和IL 10表达情况。结果与空白组比较,模型组IL 1β、IL 8、TNF α明显升高（P＜001）,IL 10明显降低（P＜001）。与模型组比较,妇科千金片高、中剂量组IL 1β,IL 8、TNF α均明显降低（P＜005）,IL 10明显升高（P＜001）;低剂量组IL 8和TNF α有所降低（P＜005）。结论妇科千金片治疗慢性盆腔炎的机制可能与抑制组织促炎症因子IL 1β、IL 8、TNF α的释放以及促进组织抗炎症因子IL 10表达、调节Th1/Th2平衡有关。     

Expansion of natural killer cells in patients with head and neck cancer: detection of "noninhibitory" (activating) killer Ig-like receptors on circulating natural killer cells

BACKGROUND: In a group of patients with head and neck cancers (H&NC), the expansion of the population of CD3-,CD16+ natural killer (NK) cells in the peripheral blood was studied. METHODS: Cytofluorimetric analysis of the expression of killer Ig-like receptors (KIR, namely p58.1, p58.2, p58.3, p70, and p140) and CD94-NKG2a was performed. Cytolytic activities were studied using 51Cr release assay. T and NK cell cloning was performed using limiting dilution culture conditions. Cytokine production was analyzed using commercial enzyme immunoassays. RESULTS: Phenotypic analysis showed that the expanded populations were heterogeneous. Even in the presence of a large number of circulating NK cells, "nonspecific" cytolytic capacities were heavily reduced, whereas cytolytic capacity related to T cells was virtually normal. Unlike NK cell clones derived from healthy donors, most NK cells derived from H&NC patients expressed surface "activating" NK cell receptors (KAR) for HLA, detected by use of a redirected cytolytic assay. Analysis of the CD4+ subpopulation at the clonal level demonstrated that they had a severe proliferative defect. CONCLUSION: These experimental data indicated that H&NC patients have a polyclonal expansion of functionally deficient NK cells expressing KAR. In addition, the proliferative capacity of patients' "helper" cells was strongly inhibited, thus accounting for a severe impairment of cytolytic activity of the expanded NK cells.     

Induction of cytokine tolerance requires internalization of Chylomicron-Bound LPS into hepatocytes

BACKGROUND: Chylomicron-bound LPS (CM-LPS) renders hepatocytes unresponsive to stimulation by proinflammatory cytokines, a process termed cytokine tolerance. We have shown that cytokine tolerance is a time- and dose-dependent process requiring functional low-density lipoprotein receptors (LDLR). Thus, we hypothesized that cytokine tolerance directly correlates with the internalization of CM-LPS complexes, and inhibition of lipoprotein binding and/or internalization inhibits the induction of cytokine tolerance in hepatocytes. MATERIALS AND METHODS: We correlated the rate of internalization of radioiodinated CM-LPS complexes with hepatocellular NO production as a measure of cytokine responsiveness. In additional studies, we used four different strategies to inhibit binding/internalization of CM-LPS via LDLR and then determined the effect of each strategy on the induction of cytokine tolerance. RESULTS: There was a strong inverse correlation between the internalization of CM-LPS and the responsiveness of hepatocytes to proinflammatory cytokines (r(2) = -0.997). Furthermore, the greater the degree of LDLR inhibition, the less susceptible hepatocytes were to the induction of cytokine tolerance by CM-bound LPS. Accordingly, cytokine tolerance induction was inhibited in hepatocytes with decreased membrane expression of LDLR as compared to control cells (69 versus 12% control; P = 0.005). Competitive inhibition of CM-LPS binding prevented internalization of CM-LPS and resulted in loss of the cytokine-tolerant phenotype. Whereas CM-LPS successfully induced cytokine tolerance in ldlr(-/-) hepatocytes, it only occurred after a prolonged pretreatment period of 8 h. CM-LPS complexes containing apolipoprotein (apo) E(2) also required a prolonged pretreatment period to induce a level of cytokine tolerance comparable to that induced by CM-LPS complexes containing either apo E(3) or E(4). CONCLUSION: Lipoprotein-bound LPS inhibits the responsiveness of hepatocytes to proinflammatory cytokines in a manner directly correlated with the internalization of LPS. Furthermore, inhibition of lipoprotein binding/internalization prevents this LPS-mediated induction of cytokine tolerance in rodent hepatocytes.     

Intestinal graft versus native liver cytokine expression in a rat model of intestinal transplantation with and without donor-specific cell augmentation

BACKGROUND: Immunomodulatory strategies such as donor-specific bone marrow or blood transfusions have been used to promote engraftment after intestinal transplants. We previously showed that delivery of donor antigen via the portal vein can effectively reduce the rate of intestinal graft rejection. The purpose of our current study was to investigate the impact of donor-specific cell augmentation (blood versus bone marrow) via the portal vein on cytokine expression in intestinal grafts versus native livers. MATERIAL AND METHODS: We performed heterotopic small intestinal transplants between male Brown-Norway (donor) and female Lewis (recipient) rats. We studied 10 groups according to the type of donor-specific cell augmentation and the use and dose of immunosuppressive therapy. For cell augmentation, donor-specific blood or bone marrow was transfused via the donor portal vein immediately before graft implantation. For immunosuppression, tacrolimus was used post-transplant at a high or low dose. Control rats received neither immunosuppression nor cell augmentation. Tissue samples for histological assessment were obtained at designated time points. RNA was extracted from intestinal graft and native liver biopsies for cytokine measurements (IL-1 alpha, IL-1 beta, IL-2, IL-4, IL-5, IL-6, IFN-gamma, TNF-alpha, and TNF-beta). Chimerism levels were determined using Q-PCR analysis. RESULTS: Without concurrent immunosuppression, neither portal donor-specific blood nor bone marrow transfusion reduced the rate of rejection. With immunosuppression, outcome was significantly better after portal donor-specific blood (versus bone marrow) transfusion. Irrespective of the type of donor-specific cell augmentation, severe rejection caused strong cytokine expression in the grafts of IL-1 alpha, IL-1 beta, IFN-gamma, and TNF-alpha; in the native livers, mainly of TNF-alpha (with IFN-gamma showing hardly any increase). In general, rejection caused stronger cytokine expression in the grafts than in the native livers. Mild rejection correlated well with strong intragraft expression of IL-6, TNF-alpha, and TNF-beta (early rejection markers); severe rejection with IL-1 alpha, IL-1 beta, IFN-gamma, and TNF-alpha (late rejection markers).In addition to cell augmentation per se, the type of cell augmentation also had an impact on cytokine expression in both grafts and native livers. Cell-augmented (versus tacrolimus-treated) rats showed hardly any differences in intragraft cytokine expression, but the expression of almost all cytokines was significantly stronger in the native livers. With immunosuppression, bone marrow infusion increased intragraft cytokine expression of IL-1 alpha, IL-1 beta, IFN-gamma, and TNF alpha, as well as liver cytokine expression of IL-1 beta, compared to blood transfusion. This finding reflected the more advanced rejection stages in the bone marrow infused group; different types of donor-specific cell augmentation had similar effects on liver cytokine expression. In the absence of myoablative therapy, chimerism levels were low, in both cell-augmented and non-cell-augmented groups. CONCLUSIONS: Rejection and donor-specific cell augmentation independently causes differences in intragraft versus native liver cytokine expression after intestinal transplants. Portal donor-specific blood transfusion, as compared with donor-specific bone marrow infusion, lowered the incidence of rejection and diminished intragraft cytokine up-regulation.     

加味当归补血汤调节IgA肾病肾虚血瘀证患者细胞因子的临床研究

目的研究加味当归补血汤对Ig A肾病患者的抑制细胞因子聚集及延缓肾脏纤维化的保护作用。方法将经过肾活检证实的Ig A肾病患者进行随机分组治疗,对照组仅使用基础治疗,治疗组在基础治疗的基础上加用加味当归补血汤。在治疗终点应用统计学方法分析2组患者血、尿细胞因子及纤维化相关指标,并进行组间疗效差异的比较。结果治疗组和对照组均可减少24 h尿蛋白水平,治疗组由治疗前的(5.78±1.10)g下降至(1.25±0.67)g,而对照组由(5.25±1.21)g下降至(2.09±0.89)g。同时具有调节患者细胞因子和纤维化相关指标如单核细胞趋化蛋白-1(MCP-1)、巨噬细胞移动抑制因子(MIF)、转化生长因子-β1(TGF-β1)、透明质酸酶(HA)、层粘连蛋白(LN)、胶原Ⅳ(Col-Ⅳ)、Ⅲ型前胶原N端肽P(PⅢNP)等的作用疗效显著,治疗前后自身对照改善均具有统计学意义(P0.05)。2组治疗后以上数据的组间比较,除尿液LN外组间比较均有统计学意义,分析其在Ig A肾病肾脏纤维化的意义表明治疗组的疗效优于对照组。结论中药和西药治疗均对Ig A肾病具有较好的抑制细胞因子聚集及延缓肾脏纤维化的保护作用,配合中药加味当归补血汤治疗可以明显提高临床疗效。     

奥瑞凝胶对反流性食管炎大鼠模型食管组织中相关基因表达及血清相关分子含量的影响

目的:研究奥瑞凝胶对反流性食管炎大鼠模型食管组织中相关基因表达以及血清中相关分子含量的影响.方法:选择成年雄性SD大鼠作为研究对象,随机分为正常组、模型组和治疗组,模型组和治疗组建立反流性食管炎模型,治疗组给予奥瑞凝胶治疗.处死大鼠后,检测血清炎症因子含量和食管组织炎症因子、多肽类神经递质、促增殖基因的表达.结果:(1)炎症因子:与模型组比较,食管组织和血清中白介素-23、17(IL-23、IL-17)的含量在治疗组中呈降低趋势;(2)多肽类神经递质:与模型组比较,食管组织中一氧化氮(NO)、NOS、血管活性肠肽(VIP)、VIP-R1、VIP-R2含量在治疗组中呈升高趋势,P物质(SP)含量治疗组中呈降低趋势;(3)促增殖基因:与模型组比较,食管组织中c-myb、增殖细胞核抗原(PCNA)和Ki-67的mRNA和蛋白含量在治疗组中呈降低趋势.结论:奥瑞凝胶治疗有助于缓解炎症反应,调节多肽类神经递质表达,抑制食管黏膜上皮过度增殖,对反流性食管炎模型大鼠具有治疗作用.     

连花定喘片治疗支气管哮喘的疗效

 目的  评价连花定喘片治疗支气管哮喘的疗效。方法  将50只SPF级BALB/C小鼠随机分为对照组、模型组、地塞米松组、连花高剂量组、连花低剂量组,每组10只。采用卵清白蛋白＋氢氧化铝致敏,并雾化激发复制小鼠支气管哮喘模型,给药组于每次激发前30 min给药,共计7次。采用特殊气道阻力值评估气道高反应性,肺组织HE染色及肺泡灌洗液(bronchoalveolar lavage fluid,BALF)细胞分类计数评价各组小鼠气道炎症性改变,ELISA法和磁性Luminex分析法检测BALF和血清中IL-4、IL-13、INF-γ的表达。结果  地塞米松组和连花高剂量组能显著降低气道阻力(P<0.05)。地塞米松组和连花高、低剂量组BALF中的各类炎性细胞数量和IL 13的表达均明显减少(P<0.05)。血清IL-13的表达在地塞米松组明显降低(P<0.05),但在连花高、低剂量组中没有变化。各组小鼠BALF和血清中IL-4和INF-γ的表达没有统计学差异。结论  连花定喘片可缓解哮喘症状,可能与其降低Th2型细胞因子IL-13的含量,从而减轻气道炎症有关。     

肝移植术中再灌注前经门静脉放血对内环境和心肺功能的影响

目的 研究肝移植术中再灌注前放血的临床意义.方法 32例肝病患者在静脉及吸入复合全麻下行无转流原位肝移植术,分为再灌注前放血组(经门静脉放血200 ml,21例)和对照组(11例).常规麻醉监测,并放置Swan-Ganz导管监测心输出量,无肝前期、无肝期给予抑肽酶、去甲肾上腺素及多巴胺,维持无肝期平均动脉压＞70 mm Hg,心输出量指数＞2.5 L·min-1·m-2.分别于门静脉阻断即刻(T1)、门静脉开放即刻(T2)、新肝期10min(T3)、新肝期30min(T4)采集桡动脉血液测定电解质、血气及炎性细胞因子浓度(肿瘤坏死因子tumor necrosis factor alpha-alpha,TNF-α;白介素6,Interleukin-6,IL-6).各时间点分别记录心肺功能参数.结果 两组患者心律失常发生率(X2=1.73,P＞0.05)和死亡率(X2=1.12,P＞0.05)没有显著差别;各时间点血钙、血镁浓度均明显低于正常值;两组患者桡动脉血钾、TNF-α、IL-6均无显著变化,再灌注前放血对乳酸的增长没有影响;各时间点肺氧合功能、心功能参数无显著变化,组间无明显差异.新肝期30 min,两组患者均表现为乳酸、炎性因子呈增高趋势,外周血管阻力指数(systemic vascular resistance index,SVRI)显著下降.结论 再灌注前放血似乎对内环境、心肺功能影响较小.     

肝移植术中门静脉阻断前后血液成分变化的研究

目的 研究肝移植术中门静脉阻断前后血液成分变化及临床意义。方法 32例肝病患者于静吸复合全麻下行无转流原位肝移植术。常规麻醉监测，并放置Swan-Ganz导管检测心输出量，无肝前期、无肝期给予抑肽酶输注，通过输液、给予去甲肾上腺素及多巴胺，维持无肝期平均动脉压〉70mmHg，心输出量指数〉2.5L·min^-1·m^-2。分别于门静脉阻断即刻、门静脉开放即刻采集门静脉血液测定电解质、血气及炎性细胞因子浓度（肿瘤坏死因子tumor necrosis factor alpha，TNF-α；白介素Interleukin-6，IL-6）。结果 阻断前、后血钙、血镁浓度均明显低于正常值。无肝期后门静脉血钾、乳酸、TNF-α及IL-6浓度显著升高（P〈0.05）；门静脉血血氧分压显著降低（P〈0.05），但仍超过40mm Hg。与C级患者相比，A级患者门静脉阻断前后血乳酸、IL-6的净变化量增加；门静脉血氧分压、二氧化碳分压净变化值呈下降趋势。结论 无肝期门静脉阻断后，其血液成分发生明显变化，开放循环前门静脉放血可能具有重要临床意义，Child A级患者门静脉开放前放血意义可能更大；但对心、肺功能影响的意义还有待探索。