紫癜性肾炎患儿外周血白三烯B4水平和CD4+CD25+调节性T细胞表达

目的 观察紫癜性肾炎(HSPN)患儿白三烯B4(LTB4)、CD4+CD25+调节性T细胞(regulatory T cell,Treg)表达及其相关性,探讨两者在过敏性紫癜(HSP)肾损害中的作用.方法 选择HSP患儿20例,HSPN患儿40例,健康对照者20例.采用ELISA法检测各组血浆LTB4水平,流式细胞术检...     

干细胞因子对内皮细胞增殖、迁移、管状形成能力的影响及对CD133+细胞的趋化效应

目的 探讨干细胞因子(SCF)对脐静脉内皮细胞(HUVEC)增殖、迁移、管状形成能力的影响,以及对CD133+细胞的趋化效应.方法 应用MTT及CCK-8增殖分析法检测HUVEC在不同细胞因子[空白试剂、SCF、血管内皮生长因子(VEGF)、抗人SCF、人IgG]条件下增殖能力的差异性;采用细胞划痕法与Matrigel体外三维成型法分别检测内皮细胞的增殖、迁移和管状形成能力;并应用Transwell技术检测不同细胞因子诱导的CD133+细胞体外趋化效应.结果 MTT及CCK-8增殖分析结果显示SCF无HUVEC增殖刺激活性;SCF可显著提升HUVEC迁移能力;SCF呈剂量依赖性增强HUVEC 管状形成能力,在适宜浓度SCF(100 ng/ml)作用下,HUVEC完整小管形成数量[(30.0 ±3.4)/105HUVEC]显著高于空白试剂组[(5.0±2.6)/105HUVEC,P<0.01];SCF可高效诱导CDl33+细胞体外趋化,SCF组[(118.0±6.5)/104CD133+细胞]Transwell小室跨膜迁移细胞数显著高于空白试剂组[(47.0±4.7)/104CDl33+细胞,P<0.01].结论 SCF可显著增强HUVEC的迁移及管状形成能力,并有效诱导CD133+细胞体外趋化,提示SCF/c-kit信号转导在内皮细胞及其祖细胞的血管新生与血管发生过程中可能发挥重要作用.

Abstract：

Objective To explore the effects of stem cell factor (SCF) on proliferation, transmigration, capillary tube formation of human umbilical vein endothelial cells (HUVEC) and on the chemotaxis of CD133+ cells. Methods In the presence of blank control, SCF, vascular endothelial growth factor ( VEGF) , anti-human SCF (anti-SCF) or human IgG, the difference in proliferation capacity of HUVEC was analyzed by MTT and CCK-8 methods, and wound scratch assay and three-diamensional in vitro Matrigel assay were used for transmigration and capillary tube formation of HUVEC, respectively. In addition, the chemotaxis of CD133 + cells sorted from human umbilical cord blood by flow cytometry was investigated by Transwell migration assay. Results SCF didn't improve the proliferative capacity of HUVEC, but significantly enhanced the transmigration capacity, and increased capillary tube formation in a dose-dependent manner.The number of intact tubules [(30.0 ±3.4)/105 HUVEC] formed by HUVECs in the presence of the optimal concentration of SCF (100 ng/ml) was remarkably higher than that in blank control group [(5.0 ±2.6)/105HUVEC,P <0.01]. SCF also significantly induced a chemotactic response of CD 133+ cells, the transmembrane migration cell number into Transwell lower chamber was significantly higher in SCF group [(118.0 ±6.5)/104 CD133+ cells] than in blank control group [(47. 0 ±4. 7)/104 CD133 + cells,P <0.01]. Conclusions SCF significantly promotes the transmigration and capillary tube formation of HUVEC, and induces a chemotactic response of CD133 + cells. SCF/c-kit signaling possibly plays a critical role in regulating angiogenesis of vascular endothelial cells and vasculogenesis of endothelial progenitor cells.     

脓毒症患者自然调节性T细胞及可溶性CD25水平研究

目的 通过前瞻性研究探讨脓毒症患者外周血自然调节性T细胞(Treg)百分比和血浆可溶性CD25分子(IL-2sRa)水平的变化及其临床价值.方法 将2009年2月至2010年2月上海交通大学附属瑞金医院重症医学科所收治的符合ACCP/SCCM于1997年提出的SIRS与脓毒症诊断标准的37例脓毒症患者与15例非感染SIRS患者,以及上海交通大学附属瑞金医院体检中心24例排除感染及免疫系统基础疾病患者,根据诊断标准分为脓毒症组、非感染SIRS组与正常对照组.其中脓毒症组男性26例,女性11例,年龄(61.67±11.87)岁;非感染SIRS组男性8例,女性7例,年龄(67.06±12.57)岁;正常对照组男14例,女10例,年龄(56.54±6.37)岁.脓毒症组肺部感染28例,腹腔感染6例,其他4例;非感染SIRS组为择期清洁手术后24 h内无感染征象者.所有入选对象均排除免疫系统基础疾病和(或)近期(30 d内)曾使用或正在使用强效免疫抑制剂患者.流式细胞分析术检测三组外周血Treg细胞百分比,ELISA检测血浆IL-2sRa,IL-4,IFN-γ水平.所得数据采用方差分析及非参数检验的Kruskal-Wallis H方法.结果 ①Treg细胞在脓毒症组、SIRS组和正常对照组占外周血CD4+CD25+T细胞的比例分别为:(66.82±21.79)%,(51.79±21.79)%、(56.45±10.68)%,脓毒症组Treg细胞显著高于SIRS组和正常对照组(P=0.001).②与SIRS组(381.664±189.83)和对照组(164.132±56.37)相比,脓毒症组可溶性CD25分子(IL-2sRa)水平(425.619±270.12)显著增高(P=0.000).③IL-2sRa浓度与Treg细胞占CD4+CD25+T细胞的比例相关分析示:Spearman相关系数=0.390,P=0.003(P＜0.05),两者存在正相关关系.结论 Treg细胞在脓毒症患者外周血的表达水平特征性增高.而外周血IL-2sRa水平可以反映Treg细胞表达水平,也有助于简化判断脓毒症的免疫状态的方法.

Abstract：

Objective To explore the CD4 + CD25 + Foxp3 + regulatory T cell percentage and plasma levels of soluble CD25 molecules in peripheral blood of septic patients and their clinical value through prospective study. Method A total of 37 septic patients and 15 non-infectious SIRS patients, who conformed to the criteria of SIRS and sepsis which proposed by American College of Chest Physicians/Society of Critical Care Medicine Consensus Conference ( ACCP/SCCM ) in 1997, were collected in ICU of Ruijin Hospital ( Shanghai Jiaotong University) from February 2009 to February 2010. Twenty-four health people were from Medical Center of Ruijin Hospital, who were excluded infection and (or) autoimmune diseases. There were 26 male and 11 female in sepsis group, average age ( 61.67 ± 11.87 ) years old; 8 male and 7 female in SIRS group, average age (67.06 ± 12.57)years old; 14 male and 10 female in health control, average age (56.54 ± 6.37 )years old. All selected patrents were excluded the autoimmune diseases and (or) patients within recent (30 days) had used or now used immunosuppressive agents. We therefore measured the Treg cell percentage in peripheral blood by Flow Cytometry and the plasma levels of IL-2sRa, IL-4, IFN-γ by ELISA. The data were analyzed by analysis of variance or nonparametric Kruskal-Wallis H test. Results ① The percentage of CD4 + CD25 + Foxp3 + regulatory T cells among septic patients, SIRS patients, and control group was: ( 66.82 ± 21.79 ) %, ( 51.79 ± 21.79 ) %, ( 56.45 ± 10. 68 ) %, respectively. septic patients showed the highest percentages of CD4 + CD25 + Foxp3 + regulatory T cell among CD4 + CD25 + T cells(P ＜ 0.05 ). ② The plasma levels of soluble CD25 in septic patients (425. 619 ± 270.12 ) were significantly higher than SIRS patients (381. 664 ± 189.83) and the control group ( 164. 1 32 ± 56.37 ) ( P ＜ 0.05 ). ③ The correlation analysis between the concentration of soluble CD25 molecules in plasma and the ratio of CD4 + CD25 + Foxp3 + regulatory T cells to CD4 + CD25 + T cells showed Spearman correlation coefficient =0.390, P = 0.003 ( P ＜ 0.05 ). Conclusion: the expression of natural regulatory T cells characteristically increased in septic patients. And the levels of soluble CD25 in peripheral blood were related to the percentages of natural regulatory T cells, which simplified the assessment of the immune status in Septic patients.     

多发性硬化患者外周血CD8+CD28-调节性T细胞的变化

Objective To investigate the levels of peripheral blood CD8+ CD28- regulatory lymphocytes and their clinical values in the patients with multiple sclerosis (MS). Method From October 2005 to August 2008, 51 patients with active rehpsing-remitting MS were enrrolled from Department of Neurology of the First Affil-iated Hospital of Wenzliou Medical College. The diagnostic criteria for MS were the 2005 revisions to the "McDon-ald criteria". All the admitted patients received 1 g of methylprednisoione per day intravenously for 5 days, fol-lowed by 60 mg prednisone per day orally for 12 days,and tapered in 6 weeks. Fourteen patients were reevaluated after corticosteroid therapy. Twenty healthy individuals ,as normal controls,matched for age and sex with the MS patients were also enrolled in this study. The percentages of peripheral blood T cells (CD8+ CD28-, CD8+CD28+, CD8+, CD4+ CD8-) were measured by flow cytometric analysis. Parametric statistical analysis were per-formed using standard methods, and linear regression analysis was conducted using Pearson correlation test. Re-sults (1)Compared with controls,the patients with active MS had significantly lower percentage of CD8+ CD28-T cells [(18.48±9.89)% vs. (24.48±4.86)%, P <0.01], and higher percentage of CD8+ CD28+ T cells [(12.23±4.31) % vs. (8.55±3.49) %, P <0.01]. (2)The percentage of CD8+ CD28- T cells was negative-ly correlated with that of CD4+ CDS- T cells (r = -0.488, P < 0.01). (3) After corticosteroid therapy, the per-eentage of peripheral blood CD8+ CD28- / CD8+ CD28+ T cells didn' t significantly decrease or increase in 14 ac-tive MS patients (P > 0.05). Conclusions The decrease of peripheral blood CD8+ CD28- regulatory T cells might be associated with the pathogenesis of MS, and CD8+ CD28- regulatory T cells perhaps played their roles through CD4+ T cells. Corticosteroid therapy could not reverse the levels of CD8+ CD28- T cells.     

多发性硬化患者外周血CD8+CD28-调节性T细胞的变化

Objective To investigate the levels of peripheral blood CD8+ CD28- regulatory lymphocytes and their clinical values in the patients with multiple sclerosis (MS). Method From October 2005 to August 2008, 51 patients with active rehpsing-remitting MS were enrrolled from Department of Neurology of the First Affil-iated Hospital of Wenzliou Medical College. The diagnostic criteria for MS were the 2005 revisions to the "McDon-ald criteria". All the admitted patients received 1 g of methylprednisoione per day intravenously for 5 days, fol-lowed by 60 mg prednisone per day orally for 12 days,and tapered in 6 weeks. Fourteen patients were reevaluated after corticosteroid therapy. Twenty healthy individuals ,as normal controls,matched for age and sex with the MS patients were also enrolled in this study. The percentages of peripheral blood T cells (CD8+ CD28-, CD8+CD28+, CD8+, CD4+ CD8-) were measured by flow cytometric analysis. Parametric statistical analysis were per-formed using standard methods, and linear regression analysis was conducted using Pearson correlation test. Re-sults (1)Compared with controls,the patients with active MS had significantly lower percentage of CD8+ CD28-T cells [(18.48±9.89)% vs. (24.48±4.86)%, P <0.01], and higher percentage of CD8+ CD28+ T cells [(12.23±4.31) % vs. (8.55±3.49) %, P <0.01]. (2)The percentage of CD8+ CD28- T cells was negative-ly correlated with that of CD4+ CDS- T cells (r = -0.488, P < 0.01). (3) After corticosteroid therapy, the per-eentage of peripheral blood CD8+ CD28- / CD8+ CD28+ T cells didn' t significantly decrease or increase in 14 ac-tive MS patients (P > 0.05). Conclusions The decrease of peripheral blood CD8+ CD28- regulatory T cells might be associated with the pathogenesis of MS, and CD8+ CD28- regulatory T cells perhaps played their roles through CD4+ T cells. Corticosteroid therapy could not reverse the levels of CD8+ CD28- T cells.     

多发性硬化患者外周血CD8+CD28-调节性T细胞的变化

Objective To investigate the levels of peripheral blood CD8+ CD28- regulatory lymphocytes and their clinical values in the patients with multiple sclerosis (MS). Method From October 2005 to August 2008, 51 patients with active rehpsing-remitting MS were enrrolled from Department of Neurology of the First Affil-iated Hospital of Wenzliou Medical College. The diagnostic criteria for MS were the 2005 revisions to the "McDon-ald criteria". All the admitted patients received 1 g of methylprednisoione per day intravenously for 5 days, fol-lowed by 60 mg prednisone per day orally for 12 days,and tapered in 6 weeks. Fourteen patients were reevaluated after corticosteroid therapy. Twenty healthy individuals ,as normal controls,matched for age and sex with the MS patients were also enrolled in this study. The percentages of peripheral blood T cells (CD8+ CD28-, CD8+CD28+, CD8+, CD4+ CD8-) were measured by flow cytometric analysis. Parametric statistical analysis were per-formed using standard methods, and linear regression analysis was conducted using Pearson correlation test. Re-sults (1)Compared with controls,the patients with active MS had significantly lower percentage of CD8+ CD28-T cells [(18.48±9.89)% vs. (24.48±4.86)%, P <0.01], and higher percentage of CD8+ CD28+ T cells [(12.23±4.31) % vs. (8.55±3.49) %, P <0.01]. (2)The percentage of CD8+ CD28- T cells was negative-ly correlated with that of CD4+ CDS- T cells (r = -0.488, P < 0.01). (3) After corticosteroid therapy, the per-eentage of peripheral blood CD8+ CD28- / CD8+ CD28+ T cells didn' t significantly decrease or increase in 14 ac-tive MS patients (P > 0.05). Conclusions The decrease of peripheral blood CD8+ CD28- regulatory T cells might be associated with the pathogenesis of MS, and CD8+ CD28- regulatory T cells perhaps played their roles through CD4+ T cells. Corticosteroid therapy could not reverse the levels of CD8+ CD28- T cells.     

多发性硬化患者外周血CD8+CD28-调节性T细胞的变化

Objective To investigate the levels of peripheral blood CD8+ CD28- regulatory lymphocytes and their clinical values in the patients with multiple sclerosis (MS). Method From October 2005 to August 2008, 51 patients with active rehpsing-remitting MS were enrrolled from Department of Neurology of the First Affil-iated Hospital of Wenzliou Medical College. The diagnostic criteria for MS were the 2005 revisions to the "McDon-ald criteria". All the admitted patients received 1 g of methylprednisoione per day intravenously for 5 days, fol-lowed by 60 mg prednisone per day orally for 12 days,and tapered in 6 weeks. Fourteen patients were reevaluated after corticosteroid therapy. Twenty healthy individuals ,as normal controls,matched for age and sex with the MS patients were also enrolled in this study. The percentages of peripheral blood T cells (CD8+ CD28-, CD8+CD28+, CD8+, CD4+ CD8-) were measured by flow cytometric analysis. Parametric statistical analysis were per-formed using standard methods, and linear regression analysis was conducted using Pearson correlation test. Re-sults (1)Compared with controls,the patients with active MS had significantly lower percentage of CD8+ CD28-T cells [(18.48±9.89)% vs. (24.48±4.86)%, P <0.01], and higher percentage of CD8+ CD28+ T cells [(12.23±4.31) % vs. (8.55±3.49) %, P <0.01]. (2)The percentage of CD8+ CD28- T cells was negative-ly correlated with that of CD4+ CDS- T cells (r = -0.488, P < 0.01). (3) After corticosteroid therapy, the per-eentage of peripheral blood CD8+ CD28- / CD8+ CD28+ T cells didn' t significantly decrease or increase in 14 ac-tive MS patients (P > 0.05). Conclusions The decrease of peripheral blood CD8+ CD28- regulatory T cells might be associated with the pathogenesis of MS, and CD8+ CD28- regulatory T cells perhaps played their roles through CD4+ T cells. Corticosteroid therapy could not reverse the levels of CD8+ CD28- T cells.     

多发性硬化患者外周血CD8+CD28-调节性T细胞的变化

Objective To investigate the levels of peripheral blood CD8+ CD28- regulatory lymphocytes and their clinical values in the patients with multiple sclerosis (MS). Method From October 2005 to August 2008, 51 patients with active rehpsing-remitting MS were enrrolled from Department of Neurology of the First Affil-iated Hospital of Wenzliou Medical College. The diagnostic criteria for MS were the 2005 revisions to the "McDon-ald criteria". All the admitted patients received 1 g of methylprednisoione per day intravenously for 5 days, fol-lowed by 60 mg prednisone per day orally for 12 days,and tapered in 6 weeks. Fourteen patients were reevaluated after corticosteroid therapy. Twenty healthy individuals ,as normal controls,matched for age and sex with the MS patients were also enrolled in this study. The percentages of peripheral blood T cells (CD8+ CD28-, CD8+CD28+, CD8+, CD4+ CD8-) were measured by flow cytometric analysis. Parametric statistical analysis were per-formed using standard methods, and linear regression analysis was conducted using Pearson correlation test. Re-sults (1)Compared with controls,the patients with active MS had significantly lower percentage of CD8+ CD28-T cells [(18.48±9.89)% vs. (24.48±4.86)%, P <0.01], and higher percentage of CD8+ CD28+ T cells [(12.23±4.31) % vs. (8.55±3.49) %, P <0.01]. (2)The percentage of CD8+ CD28- T cells was negative-ly correlated with that of CD4+ CDS- T cells (r = -0.488, P < 0.01). (3) After corticosteroid therapy, the per-eentage of peripheral blood CD8+ CD28- / CD8+ CD28+ T cells didn' t significantly decrease or increase in 14 ac-tive MS patients (P > 0.05). Conclusions The decrease of peripheral blood CD8+ CD28- regulatory T cells might be associated with the pathogenesis of MS, and CD8+ CD28- regulatory T cells perhaps played their roles through CD4+ T cells. Corticosteroid therapy could not reverse the levels of CD8+ CD28- T cells.     

多发性硬化患者外周血CD8+CD28-调节性T细胞的变化

Objective To investigate the levels of peripheral blood CD8+ CD28- regulatory lymphocytes and their clinical values in the patients with multiple sclerosis (MS). Method From October 2005 to August 2008, 51 patients with active rehpsing-remitting MS were enrrolled from Department of Neurology of the First Affil-iated Hospital of Wenzliou Medical College. The diagnostic criteria for MS were the 2005 revisions to the "McDon-ald criteria". All the admitted patients received 1 g of methylprednisoione per day intravenously for 5 days, fol-lowed by 60 mg prednisone per day orally for 12 days,and tapered in 6 weeks. Fourteen patients were reevaluated after corticosteroid therapy. Twenty healthy individuals ,as normal controls,matched for age and sex with the MS patients were also enrolled in this study. The percentages of peripheral blood T cells (CD8+ CD28-, CD8+CD28+, CD8+, CD4+ CD8-) were measured by flow cytometric analysis. Parametric statistical analysis were per-formed using standard methods, and linear regression analysis was conducted using Pearson correlation test. Re-sults (1)Compared with controls,the patients with active MS had significantly lower percentage of CD8+ CD28-T cells [(18.48±9.89)% vs. (24.48±4.86)%, P <0.01], and higher percentage of CD8+ CD28+ T cells [(12.23±4.31) % vs. (8.55±3.49) %, P <0.01]. (2)The percentage of CD8+ CD28- T cells was negative-ly correlated with that of CD4+ CDS- T cells (r = -0.488, P < 0.01). (3) After corticosteroid therapy, the per-eentage of peripheral blood CD8+ CD28- / CD8+ CD28+ T cells didn' t significantly decrease or increase in 14 ac-tive MS patients (P > 0.05). Conclusions The decrease of peripheral blood CD8+ CD28- regulatory T cells might be associated with the pathogenesis of MS, and CD8+ CD28- regulatory T cells perhaps played their roles through CD4+ T cells. Corticosteroid therapy could not reverse the levels of CD8+ CD28- T cells.     

60Co γ射线致死剂量全身照射对小鼠脾脏和肠系膜淋巴结T细胞亚群表面免疫分子表达水平的影响

目的 探讨小鼠接受60Co γ射线致死剂量全身照射(TBI)后,脾脏和肠系膜淋巴结T细胞活化分子和黏附分子表达水平的变化.方法 于2012年7月至10月,选择12只无特定病原体(SPF)级雄性BALB/c小鼠为研究对象.研究对象纳人标准:周龄为10～12周龄,体重为20～25 g.按照完全随机法将其分为TBI组(接受致死剂量TBI)和对照组(未照射),每组6只.TBI组予60C0 γ放射源一次性照射(剂量率为0.66 Gy/min,总剂量为7.5 Gy).于照射后第7天,脊椎离断处死两组小鼠,无菌条件下,取其脾脏、肠系膜淋巴结,研磨后采用密度梯度离心法收集单个核细胞.采用流式细胞术检测两组小鼠脾脏和肠系膜淋巴结中,CD4+T,CD8+T,辅助性T细胞(Th)1及细胞毒性T细胞(Tc)1细胞表面CD44和CD62L的表达水平.并采用统计学方法比较两组小鼠上述免疫分子表达水平.结果 照射后第7天,TBI组小鼠脾脏和肠系膜淋巴结CD44+ CD4+T细胞比例为(88.1±1.1)％和(76.7±1.8)％,均低于对照组的(95.0±1.1)％和(92.8±1.4)％,且差异有统计学意义(t=-11.01、-17.82,P＜0.05);而CD44+ CD8+T细胞的比例分别为(99.0±0.3)％和(92.5±1.7)％,均高于对照组的(88.7±0.8)％和(83.4±1.6)％,差异亦有统计学意义(t=31.15、9.54,P＜0.05).TBI组小鼠肠系膜淋巴结CD44 Th1比例为(4.1±0.4)％,高于对照组的(1.0±0.2)％,且差异有统计学意义(t=14.78,P＜0.05);而两组小鼠脾脏CD44+Th1比例比较,差异无统计学意义(t=-0.93,P＞0.05).TBI组小鼠脾脏和肠系膜淋巴结CD44+ Tc1比例分别为(65.5±4.6)％和(26.6±2.8)％,较对照组的(41.8±1.5)％和(18.7±1.3)％增高,且差异有统计学意义(t=9.97、6.51,P＜0.05).TBI组小鼠的脾脏和肠系膜淋巴结中的CD4+T和CD8+T细胞表面CD62L的表达水平均低于对照组,脾脏中表达水平分别为(0.3±0.0)％比(66.7±4.7)％,(1.1±0.2)％比(64.4±5.1)％;肠系膜淋巴结中为(2.6±0.3)％比(89.7±2.8)％,(9.1±0.5)％比(79.4±3.3)％,两组上述4项指标比较,差异均有统计学意义(t=-24.52、-21.53、-53.75、-36.56,P＜0.05).结论 小鼠接受TBI后,脾脏和肠系膜淋巴结的T细胞被激活,表现为活化分子CD44在效应T细胞表达升高,而黏附分子CD62L表达下调,T细胞表型改变.通过检测T细胞表面CD44、CD62L等活化及黏附分子的表达水平,可在一定程度上反映体内T细胞活化情况,为评估TBI预处理方案应用后,机体T细胞功能状态的改变提供依据.     

ICU危重症患者免疫功能的研究

目的 观察危重症患者T淋巴细胞亚群、NK细胞活性、免疫球蛋白、白蛋白和补体水平,了解危重症患者的免疫功能状态.方法 观察20例ICU的危重症患者(APACHEⅡ评分≥16分)和20例健康体检者的细胞免疫、体液免疫和白蛋白水平.结果 ①危重症患者CD3、CD4、NK细胞活性和IgM、补体C3水平比健康者明显降低[(55.41±21.09)%与(73.93±5.77)%;(20.87±8.16)%与(39.04±4.24)%;(11.56±6.65)%与(16.94±4.55)%;(0.88±0.43)%与(1.32±0.50)%;(0.91±0.36)g/L与(1.31±0.44)g/L;均P＜0.01];IgA水平升高[(3.02±1.23)g/L与(2.05±0.71)g/L,P＜0.01];②白蛋白和CD4细胞活性APACHEⅡ评分≤25分患者明显高于APACHE Ⅱ评分＞25分的患者[(26.99±3.98)g/L与(20.39±3.30)g/L;(25.38±6.05)%与(17.36±6.17)%;均P＜0.05];③外周血T淋巴细胞CD4细胞活性和白蛋白水平与APACHEⅡ评分负相关r=-0.476,P=0.0335;r=-0.6659,P=0.0014).结论 危重症患者细胞免疫功能和体液免疫功能受到抑制,外周血CD4细胞活性可以反映危重症患者免疫功能.     

原发免疫性血小板减少症患儿中T淋巴细胞亚群、NK细胞及B细胞在外周血中的表达情况分析

目的研究原发免疫性血小板减少症患儿中T淋巴细胞亚群、NK细胞及B细胞在外周血中的表达情况。方法选择ITP患儿241例作为研究组,另选择252例健康儿童作为对照组,检测并比较2组患儿T淋巴细胞亚群(CD_3、CD_4、CD_8细胞)、B细胞和NK细胞水平。结果研究组外周静脉血中CD_3水平、CD_4水平和CD_4/CD_8显著低于对照组,CD_8水平显著高于对照组(P0.05);研究组血清中B细胞水平低于对照组。研究组的PAIgG、PAIgA、PAIgM水平显著高于对照组(P0.05),IFN-γ水平高于对照组,IL-4水平低于对照组;研究组CD(16+56)水平高于对照组。结论 ITP患儿机体存在T淋巴细胞水平失衡,NK细胞以及B细胞表达未见异常发生。     

重型再生障碍性贫血患者CD8+效应T细胞损伤骨髓造血途径的研究

目的 研究重型再生障碍性贫血(SAA)患者外周血CD8+ CD25+和CD8+HLA-DR+T细胞数量及其杀伤靶细胞的途径,探讨SAA的免疫发病机制。方法 应用流式细胞术检测29例SAA(初治14例、缓解15例)患者及12名健康对照者外周血CD8+ CD25+和CD8+ HLA-DR+T细胞的数量及其胞质内穿孔素、颗粒酶B、肿瘤坏死因子β(TNF-β)、胞膜FasL表达。结果 SAA初治组患者CD8+ CD25+T细胞占CD8+和CD3+T细胞的比例分别为(3.67±2.58)％和(2.25±1.35)％,缓解组为(5.19±4.29)％和(2.98±1.35)％,正常对照组为(4.84±2.31)％和(2.11±1.88)％,CD8+CD25+T细胞占CD8+和CD3+T细胞的比例三组间比较差异无统计学意义(P值均＞0.05)。初治组CD8+ HLA-DR+T细胞占CD8+T细胞比例为(39.30±8.13)％,缓解组为(20.65±5.38)％,正常对照组为(18.34±6.68)％,初治组明显高于缓解组及正常对照组(P＜0.01),缓解组与正常对照组比较差异无统计学意义(P＞0.05)。SAA初治组CD8+ HLA-DR+T细胞占CD3+T细胞比例为(27.81±7.10)％,缓解组为(12.02±3.03)％,正常对照组为(8.50±2.33)％,初治组明显高于缓解组及正常对照组(P＜0.01),缓解组明显高于正常对照组(P＜0.05)。SAA初治组CD8+ HLA-DR+T细胞穿孔素、颗粒酶、TNF- β、FasL中位表达比例分别为8.51％、96.08％、72.11％、94.25％,均明显高于缓解组(1.78％、85.20％、34.38％、51.20％)及正常对照组(1.86％、82.09％、17.92％、32.91％)(P值均＜0.05),缓解组与正常对照组比较差异无统计学意义(P＞0.05)。结论 SAA患者外周血CD8+ HLA-DR+效应T细胞比例增加,CD8+效应T细胞影响靶细胞的穿孔素-颗粒酶途径、细胞因子(TNF-β)途径、Fas/FasL途径均参与了骨髓造血细胞损伤的过程。     

不同营养途径对食管癌术后细胞免疫功能的影响

目的 探讨不同营养途径对食管癌术后细胞免疫功能的影响.方法 将62例食管癌手术治疗患者应用随机数字表随机分为早期肠内营养（EN）组和肠外营养（PN）组,各31例;分别于术前及术后第1、8天进行细胞免疫指标（T淋巴细胞及其亚群、自然杀伤细胞、B淋巴细胞）检测,两组计量资料的比较采用t检验,两组多个时间点计量资料的比较采用重复测量资料的方差分析.结果 EN组术前CD3、CD4、CD8、CD4/CD8、自然杀伤细胞和B淋巴细胞分别为（60.46±6.41）％、（39.03±5.64）％、（25.06±5.39）％、1.60±0.24、（13.86±4.91）％、（13.14±3.81）％,术后第8天分别为（65.18±7.81）％、（40.45±6.82）％、（22.81±4.47）％、1.78±0.16、（16.97 ±4.85）％、（15.69±3.88）％,与术前比较差异均有统计学意义（P均＜0.05）;PN组术前CD3、CD4、CD8、CD4/CD8、自然杀伤细胞和B淋巴细胞分别为（59.78±5.17）％、（39.10±5.49）％、（24.94±3.97）％、1.57±0.20、（14.02±4.47）％、（12.96±3.39）％,术后第8天分别为（60.27±6.20）％、（39.71±5.63）％、（24.74±3.80）％、1.62±0.15、（15.35±4.31）％、（14.33±3.43）％,与术前比较除CD3、CD8外差异均有统计学意义;EN组与PN组术后第8天比较除CD4外差异均有统计学意义（P均＜0.05）.结论 食管癌术后早期EN较PN可显著改善患者的细胞免疫功能.     

严重烧伤患者外周血树突细胞的变化及临床意义

目的 探讨严重烧伤患者外周血树突细胞(DC)的变化及与烧伤严重程度和脓毒症发生的关系.方法 22例严重烧伤患者根据脓毒症的诊断分为烧伤组(10例)和烧伤脓毒症组(12例);按烧伤总体表面积(TBSA)分为TBSA Ⅰ组(TBSA 30％～50％,14例)和TBSA Ⅱ组(TBSA 51％～80％,8例).采集患者伤后1、3、7、14和20 d外周静脉血,用流式细胞仪检测外周血中髓样树突细胞(mDC,Lineage1- HLA-DR+CD11e+)和浆样树突细胞(pDC,Lineage1-HLA-DR +CD123+)两种DC亚型的数量.以同期10例健康体检者作为对照.结果健康对照组外周血mDC为(0.450±0.150)％,pDC为(0.241±0.084)％.与健康对照组比较,烧伤组患者伤后1、3、7d外周血mDC[(0.257±0.116)％、(0.274±0.086)％、(0.317±0.056)％]和pDC[(0.122±0.058)％、(0.165±0.051)％、(0.177±0.024)％]均显著减少(均P＜0.05),14 d、20 d时mDC、pDC数量恢复至正常水平;烧伤脓毒症组患者伤后1d外周血mDC[(0.230±0.090)％]和pDC[(0.114±0.071)％]即较烧伤组进一步减少(均P＜0.05),至20 d时外周血mDC和pDC仍显著低于烧伤组[mDC:(0.246±0.076)％比(0.412±0.097)％; pDC:(0.097±0.032)％比(0.203±0.039)％,均P＜0.05].与健康对照组比较,TBSA I组患者伤后1、3、7d外周血mDC[(0.266±0.062)％、(0.289±0.071)％、(0.351±0.054)％]和pDC[(0.131±0.025)％、(0.163±0.037)％、(0.178±0.038)％]均显著减少(均P＜0.05),14 d、20 d时mDC、pDC数量恢复至正常水平;TBSAⅡ组患者伤后1d外周血mDC[(0.227±0.070)％]和pDC[(o.112±0.047)％]即较TBSA Ⅰ组进一步减少(均P＜0.05),至20 d外周血mDC和pDC仍显著低于TBSA Ⅰ组[mDC:(0.297±0.072)％比(0.423±0.046)％;pDC:(0.107±0.061)％比(0.197±0.042)％,均P＜0.05].结论严重烧伤患者早期外周血mDC和pDC数量均减少,烧伤程度越严重DC数量减少越明显;外周血DC数量减少导致烧伤后机体免疫功能低下,数量减少明显者易并发脓毒症;烧伤后定期检测外周血DC数量可以作为脓毒症的早期预警指标.     

急性冠脉综合征患者不同部位血Th17细胞和IL-17对比分析

目的 比较急性冠状动脉综合征(acute coronary syndrome,ACS)患者周围血和冠状动脉罪犯血管病变处血辅助性T细胞17 (helper T cells,Th17)占CD4+T细胞的比例以及IL-17水平,分析其临床意义.方法 序贯入选2012年2月15 日至2012年10月15日杭州师范大学附属医院接受冠状动脉造影(CAG)的冠心病(coronary heart disease,CHD)及疑诊冠心病患者76例.分为ACS组、稳定性心绞痛(stable angina,SA)组和对照组.所有患者均在做完冠状动脉造影后即刻采集肘静脉血5 mL.ACS患者同时采集罪犯血管病变处血5 mL.采用流式细胞术检测不同部位血 Th17的百分率.用ELISA法检测不同部位血血清白细胞介素(interleukin,IL)-17的质量浓度.结果 ACS患者周围血Th17细胞占CD4+T细胞的比例和冠状动脉罪犯血管病变处血 Th17细胞占CD4+T细胞的比例分别是(3.18±0.29)％和(3.17±0.30)％,差异无统计学意义(P =0.919).ACS组、SA组和对照组外周血 Th17细胞占CD4+T细胞的比例分别是(3.18±0.29)％、(1.32±0.31)％和(1.28±0.33)％,ACS组与另外两组相比差异均有统计学意义(P＜0.01),但SA组和对照组比较,差异无统计学意义.ACS外周血和冠状动脉罪犯血管病变处血血清IL-17质量浓度分别是(81.23±18.63) pg/mL和(82.37 ±20.51) pg/mL,差异无统计学意义(P =0.573).ACS组外周血IL-17水平分别与SA组和对照组相比,差异均有统计学意义.但SA组和对照组比较差异无统计学意义.结论 ACS患者周围血 Th17细胞百分比以及IL-17水平与冠状动脉罪犯血管病变处相似,原因可能是多方面的.ACS患者外周血 Th17细胞比例增加、IL-17水平升高,这可能参与动脉粥样硬化斑块不稳定的形成,这个过程导致ACS发病.     

重组人脑利钠肽治疗顽固性心力衰竭临床研究

目的 研究重组人脑利钠肽(rhBNP)治疗顽固性心力衰竭(RHF)患者血液动力学效应及安全性.方法 选择住院期间经常规抗心力衰竭药物强化治疗无明显改善的RHF患者16例,给予rhBNP 1.5 μg/kg静脉冲击后,以0.007 5μg/(kg·min)连续静脉滴注24h.在给药后24h内监测有创血液动力学参数、血压、心率及血清生化指标,应用rhBND 3 d后,行超声心动图检测心脏指数(CI)、左心室舒张末期容积(LEVDD)、左心室射血分数(LVEF).结果 在注射rhBNP 15 min时肺毛细血管楔压[PCWP(22.7±4.0)mm Hg与(25.3±3.9)mm Hg]、平均肺动脉压[MPAP(31.9±3.6)mm Hg与(34.6±7.8)mm Hg]较基础值明显下降(P＜0.05);收缩压在给药后1h下降[(105.2±11.5)mm Hg与(119.0±17.2)mm Hg],差异有统计学意义(P均＜0.01),以后逐渐恢复至基线时水平;用药24h心率也明显改善[(109.0±10.8)次/min与(82.2±8.6)次/min](P ＜0.01);血钾、血钠、血肌酐无明显变化;用药3d后CI[(3.7±0.6)L/m2与(1.8±0.4)L/m2]和EF[(43.1±8.3)％与(31.2±6.4)％]均有明显改善,LVEDD明显缩小[(63.6±5.7)mm与(67.3±6.2)mm](P均＜0.01).未发生与rhBNP相关的症状性低血压及其他严重不良反应.结论 静脉应用rhBNP能迅速改善RHF患者血液动力学状况,且安全可行.     

56例中年女性尿失禁患者的尿流动力学分析

目的 研究中年女性(35～55岁)尿失禁患者的尿流动力学变化,提高相应诊治水平.方法 对56例尿失禁的中年女性患者进行尿流动力学研究,并与正常对照组(17例)比较.结果 56例患者中压力性(SUI)尿失禁患者为33例(58.93％)(SUI组)、急迫性(UUI)尿失禁患者为9例(16.07％)(UUI组)、混合性(SUI/UUI)尿失禁患者为14例(25.00％)(SUI/UUI组).SUI组患者最大尿流率(Qmax)(27.72±5.21)ml/s高于正常对照组(20.45±7.15)ml/s,差异有统计学意义(P＜0.05);UUI组的膀胱初感容量(FS)、正常排尿感觉(ND)、强烈排尿感觉(SD)、急迫排尿感觉(UD)与正常对照组[(135.65±42.73)ml与(132.70±40.65)ml、(166.24±51.42)ml与(160.70±50.44)ml、(315.75±42.34)ml与(320.75±42.34)ml、(320.24±45.03)ml与(335.75±51.98)ml]比较差异均有统计学意义(P均＜0.05);SUI组及SUI/UUI组患者测得Valsalva漏尿点压力(ALPP)[(62.29±25.40)cm H2O与(88.30±28.54)cm H2O]比较差异有统计学意义(t=13.041,P＜0.05);SUI组患者的最大尿流率时逼尿肌压(Pdet-Qmax)、最大尿道压(MUP)及最大尿道闭合压(MUCP)明显低于正常对照组[(24.29±6.24)cm H2O与(34.45±8.20)cm H2O、(68.20±18.27)cm H2O与(87.14±17.26)cm H2O、(74.24±35.75)cm H2O与(90.66±30.10)cm H2O]差异均有统计学意义(P均＜0.05).结论 中年女性尿失禁患者具有较大患者群体,其分类较为复杂,尿流动力学检查对于明确其诊断及选择正确治疗方法,提高疗效具有重要指导意义.     

SARS患者康复期淋巴细胞功能状态及T细胞受体Vβ24个亚家族表达的格局

背景:严重急性呼吸道综合征(severe acuterespi ratory syndrome,SARS)是由SARS相关冠状病毒感染引起的急性传染性疾病。但目前其康复期患者细胞免疫状态仍有待进一步研究。目的:观察SARS患者康复期淋巴细胞功能状态和T细胞受体(TCR)Vβ24个亚家族表达的格局。设计:前后对照观察。单位:广东省中医院中心实验室。对象:选择2003-01/04在广州中医药大学第二附属医院收治的76例治愈的SARS患者,全部病例均符合"非典型肺炎的临床诊断标准"、"非典型肺炎重症病例诊断标准、出院标准"及《中医临床诊疗术语证候部分》的诊断标准,男30例,女46例,平均(32±11)岁。选择10名同期健康到院体检者,女5名,男5名,平均(32±7)岁,所有受试对象对检测项目知情同意。方法:①TCRVβ24个亚家族的表达检测:取受试对象全血2mL,用EDTA-K2抗凝管抗凝。在流式细胞术检测专用管中,分别加入各种荧光素标记的mAb:抗CD3、抗CD4、抗CD8、抗CD25、抗CD28、抗HLA-DR、PC5-抗CD3mAb、TCRVβ1(PE FITC)、Vβ2(PE FITC)、Vβ3(FITC)、Vβ4(PE FITC)、Vβ5.1(PE FITC)、Vβ5.2(PE)、Vβ5.3(PE)、Vβ7.1(PE FITC)、Vβ7.2(FITC)、Vβ8(FITC)、Vβ9(PE)、Vβ11(PE)、Vβ12(FITC)、Vβ13.1(PE)、Vβ13.2(PE)、Vβ13.6(PE FITC)、Vβ14(FITC)、Vβ16(FITC)、Vβ17(PE FITC)、Vβ18(PE)、Vβ20(FITC)、Vβ21.3(FITC)、Vβ22(PE FITC)和Vβ23(PE)10μL,然后加入抗凝血50μL。混匀后,于室温避光孵育15min,再加入溶血素溶血、洗涤。最后将沉淀溶在300μLPBS中,用CoulterESP流式细胞仪进行检测。②T细胞亚群,T、B细胞的活化状态及Ts细胞和Tc细胞百分率的检测:每次收集5000个细胞,用仪器所带软件分别计算T细胞亚群(CD3、CD4和CD8)、活化的T、B细胞(CD3 /CD25 、CD3 /HLA-DR 、和CD3-/HLA-DR )、Ts细胞和Tc细胞(CD8 /CD28 、CD8 /CD28-)的百分率。主要观察指标:①T细胞亚群(CD3、CD4和CD8)的表达。②活化的T、B细胞(CD3 /CD25 、CD3 /HLA-DR 、和CD3-/HLA-DR )的表达。③Ts细胞和Tc细胞(CD8 /CD28 、CD8 /CD28-)的百分率。④TCRVβ24个亚家族的表达。结果:所有受试对象均进入TCRVβ24个亚家族检测结果分析,74例SARS患者及健康对照者10名均进入其余检测结果分析。①T细胞亚群的检测结果:CD4 细胞的百分率均数低于参考值[(33.33±6.64)%,(43±9)%,P<0.01],CD8 细胞的百分率高于参考值[(34.07±6.40)%,(30±9)%,P<0.01]。②活化的T、B细胞表达检测:表达HLA-DR的T、B细胞增加,而表达CD25的T细胞减少。有53例CD25 细胞的百分率低于正常参考值,有64例CD3 /CD25 T细胞的百分率低于正常参考值。只有CD3 /HLA-DR 和CD3-/HLA-DR 细胞的百分率高于正常参考值,其中表达CD3 /HLA-DR 的T细胞增加36例,表达CD3-/HLA-DR 的T细胞增加30例。③Ts细胞和Tc细胞的百分率:Ts细胞(CD8 /CD28-)的百分率较参考值增高[(28.75±7.31)%,(15.99±5.1)%,P<0.01];Tc细胞(CD8 /CD28 )的百分率则低于参考值[(5.99±3.60)%,(13.2±4.1)%,P<0.01]。其中有39例Tc细胞的百分率降低,有48例Ts细胞的百分率升高,CD4 与CD8 T细胞的比值均在正常参考值范围内。④TCRVβ24个亚家族表达:康复期SARS患者TCRVβ24个亚家族表达中,TCRVβ14的表达最高,TCRVβ5.3、和23表达次之,出现TCRVβ24个亚家族表达的不同。正常对照组中只有TCRVβ14表达最高,与SARS患者的结果相一致,但TCRVβ24个亚家族的分布格局不同。两组进行比较,Vβ1、Vβ5.2、Vβ5.3、Vβ7.2、Vβ9、Vβ11、Vβ13.1、Vβ13.2、Vβ17、Vβ18、Vβ22和Vβ23的表达差异显著,康复期SARS患者组均高于正常对照组(P<0.05-0.01)。结论:康复期SARS患者CD8 CD28-T细胞数的升高可导致CD8 T细胞数升高;而Ts细胞数的增加,因其分泌的抑制因子增多,从而造成CD3 和CD4 T细胞数降低。SARS患者康复期TCRVβ24个亚家族在T细胞上的表达不同。CD3 /HLA-DR 和CD3-/HLA-DR T细胞数的升高,可能与T、B细胞的活化较晚有关。