慢性丙型肝炎患者抗病毒治疗前后外周血树突状细胞表面共刺激分子的变化及初步分析

目的了解慢性丙型肝炎病毒(HCV)感染者外周血树突状细胞(DC)中髓样DC(mDC)和淋巴样DC(pDC)的表型频数,分析标准抗病毒治疗后获得完全早期病毒学应答(cEVR)患者DC表面共刺激分子(CD80、CD83、CD86)表达变化,探讨不同类型DC的活化在慢性HCV感染发病机制中的作用。方法选取我院住院的20例慢性HCV感染者,在接受初始聚乙二醇化干扰素-α(PEG-IFNα)联合利巴韦林抗病毒治疗达cEVR前后,分离外周血单个核细胞(PBMC),应用流式细胞仪检测mDC和pDC的频数及其表面CD80、CD83、CD86的表达水平。结果与健康对照组相比,慢性HCV感染者mDC、pDC频数均降低,其表面CD80的表达水平降低,CD86的表达水平升高,CD83的表达水平无明显差异。与治疗前相比,经干扰素和利巴韦林治疗达cEVR后,mDC上的CD80表达水平升高,CD83、CD86的表达水平降低;pDC上的CD80、CD83、CD86的表达水平均上升。结论慢性HCV感染者的外周血mDC、pDC频数降低,共刺激分子表达部分受损,干扰素抗病毒治疗可以恢复DC共刺激分子表达。提示DC数量和功能障碍可能是慢性HCV持续感染的重要机制之一。     

慢性乙型肝炎患者外周血树突状细胞上Toll样受体的表达变化

目的 分析慢性乙型肝炎患者外周血树突状细胞(DC)上Toll样受体3(TLR3)的表达变化及其临床意义.方法 随机筛选慢性乙型肝炎(CHB)患者20例,并以10例健康者作对照,分别从外周血分离单个核细胞,体外诱导培养DC.流式细胞仪(FCM)测定未成熟期DC(imDC)与成熟期DC(mDC)的表型;分别应用流式细胞仪和Western印迹法测定imDC与mDC上TLR3的表达率和受体蛋白的表达.结果 在健康对照组,mDC表面分子CD80、CD86、组织相容性白细胞抗原(HLA)-DR、CD83的表达率分别为(82.35±8.67)%、(79.61±10.08)%、(92.79±8.48)%和(83.76±5.47)%,imDC表面分子表达率分别为(28.31±8.79)%、(31.17±11.23)%、(27.61±10.28)%和(23.46±11.53)%,各组间比较,差异均有统计学意义(P＜0.01);而在CHB组,mDC与imDC表面分子表达率分别为(42.46±9.22)%、(40.72±11.24)%、(48.57±12.51)%、(22.25±11.22)%和(18.57±10.22)%、(24.16±10.46)%、(17.87±10.38)%、(14.28±6.77)%,各组间比较,差异均无统计学意义(P＞0.05);健康者组mDC上表面分子的表达率与CHB患者组比较,各组间差异均有统计学意义,而两组受试者imDC上表面分子的表达率比较,差异均无统计学意义.健康对照imDC上TLR3的表达率明显高于mDC(P＜0.01),而CHB患者imDC与mDC上TLR3的表达率差异无统计学意义(P＞0.05);且CHB患者imDC上TLR3的表达率明显低于健康对照(P＜0.01).结论 CHB患者外周血DC上存在TLR3的表达下调,可能是HBV感染慢性化的重要原因.     

慢性乙型肝炎患者外周血树突状细胞HBV DNA载量与其功能的关系

目的:探讨乙型肝炎患者外周血DCs HBV DNA载量与DCs功能的关系.方法:采集20例慢性乙型肝炎患者和10例健康人的抗凝外周静脉血,分离外周血单个核细胞(PBMCs),IL-4和GM-CSF的作用下培养使DCs增殖、成熟,以流式细胞技术分别检测DCs表面HLA-DR、CD-1α、CD80及CD86的表达:ELISA检测DCs培养上清液IL-12的水平,实时荧光定量PCR法检测DCs HBV DNA的载量,同种异体混合淋巴细胞反应检测DCs刺激同种异体T细胞增殖能力.结果:慢性乙型肝炎患者外周血DCs内亦可检测至HBv DNA,患者DCs表面HLA-DR、CD-1α、CD80及CD86的表达水平、DCs分泌IL-12水平及DCs刺激同种异体T细胞增殖能力均显著低于健康对照组,其中DCs HBV DNA阳性组较DCs HBV DNA阴性组下降更为明显(39.17±6.02 VS 60.11±7.92,46.03±5.52 VS 58.77±4.12,20.95±6.32 VS 35.24±7.41,25.16±6.05 VS 45.30±8.01,19.67±7.32VS 30.74±8.39,0.32±0.08 VS 0.59±0.11,均P＜0.01或0.05);HLA-DR、CD-1α、CD80及CD86的表达、IL-12水平及DCs刺激同种异体T细胞增殖能力与DCs HBV DNA载量呈显著负相关(r=0.713,-0.713,-0.623,-0.702,-0.525,-0.841,均P＜0.001).结论:慢性乙型肝炎患者外周血DCs可被HBV DNA感染,患者DCs HBV DNA载量与DCs功能有密切关系,病毒栽量低者DCs功能好,病毒栽量高者DCs功能差.     

慢性乙型肝炎患者外周血CD4+CD25+调节性T细胞和T淋巴细胞亚群的研究

目的 观察外周血CD4 CD25 调节性T细胞(Tregs)百分比和T淋巴细胞亚群在慢性乙型肝炎(CHB)患者中的特点.方法 采集CHB患者15例和健康对照20例的外周血单个核细胞(PBMC)标本,应用三色流式分析法对PBMC中CD4 CD25 Tregs占CD4 百分比及外周血中的T淋巴细胞亚群进行分析.结果 分别比较CD4 CD25highTregs和CD4 CD25 CD127low/-Tregs占CD4 T细胞百分比,两组表型的Tregs在CHB组均高于正常对照组[(4.25%±3.22%vs1.57%±0.64%),(4.39%±2.03%vs1.86%±0.84%)](P＜0.01).CHB组外周血中的CD4 T淋巴细胞亚群的百分比(32.28%±6.26%)较正常对照组(39.39%±10.73%)下降,统计学有差别(P＜0.05).外周血中CD8 及CD4 /CD8 比值在两组间变化无统计学差异.结论 CHB患者CD4 CD25high和CD4 CD25 CD127low/-Tregs升高,外周血中的CD4 T淋巴细胞百分比下降.     

慢性乙型肝炎患者B7-H1和PD-1信号途径表达的特点

目的 初步探讨免疫细胞协同共刺激分子途径之一--B7-H1和PD-1表达在慢性乙型肝炎(CHB)患者中的特点.方法 收集11例CHB患者和16例健康人的外周血肝素抗凝标本,流式细胞技术检测CHB患者外周血髓样树突细胞(mDC)和T淋巴细胞上B7-H1和PD-1的表达.数据比较采用t检验.结果 CHB患者外周血CD3~+T淋巴细胞、CD4~+T淋巴细胞、CD8~+T淋巴细胞和mDC上B7-H1阳性表达率分别为(40.69±14.49)%、(42.84±11.19)%、(33.48±14.07)%和(16.60±4.04)%.健康对照者分别为(14.66±10.11)%、(4.62±3.84)%、(1.89±2.31)%和(0.49±0.37)%,CHB患者B7-H1在T淋巴细胞和mDC上的表达水平均明显高于健康者,差异具有统计学意义(t=-2.884,t=-10.894,t=-7.378,t=-13.182;均P<0.05).CHB患者CD3~+T淋巴细胞、CD4~+T淋巴细胞和CD8~+T淋巴细胞上PD-1阳性表达率分别为(12.45±6.36)%、(11.42±6.20)%和(13.03±6.71)%,健康对照者分别为(7.80±3.53)%、(7.12±2.60)%和(7.88±3.74)%.CHB患者PD-1在T淋巴细胞上的表达水平也均明显高于健康者,差异具有统计学意义(t=-2.323,t=-2.355,t=-2.439;均P<0.05).结论 B7-H1和PD-1在CHB患者外周血mDC和T淋巴细胞上表达较高.     

HBsAg负载的慢性乙肝患者DCs诱导特异性Th1细胞分化的作用

目的:研究HBsAg负载的慢性乙肝患者外周血树突状细胞(dendritic cells,DCs)对自身Th1细胞分化的诱导作用.方法:Fico11密度梯度离心和贴壁法分离慢性乙肝患者外周血单核细胞,以含ThIL-4 rhGMCSF的完全培养基诱导DCs.流式细胞术检测各组DC表面CD80,CD86,CD40和HLA-DR分子的表达,CCK-8法检测各组DCs刺激同种异体淋巴细胞增殖的能力,ELISA法检测各组DCs培养液上清中IL-12的水平.免疫磁珠分选慢性乙肝患者外周血CD4 T细胞,分别与患者自身的DCs共培养,细胞内细胞因子染色,流式细胞仪检测共培养后CD4 T细胞内特征性细胞因子IFN-γ和IL-4,ELISA法检测共培养上清中IFN-γ/IL-4的水平.结果:与对照组相比,HBsAg、IFN-γ和HBsAg IFN-γ组DCs表面表达CD80,CD86,CD40和HLA-DR分子水平较高;刺激同种异体淋巴细胞增殖的能力较强;DCs分泌的上清液中、IL-12水平也较高.与对照组相比,HBsAg、IFN-γ和HBsAg IFN-γ组DCs与自身Th细胞共培养后,Th1细胞占CD4 T细胞的百分比升高(10.76%±3.98%,11.43%±4.32%,15.28%±4.73% vs 7.84%±3.10%,P＜0.01),Th2细胞占CD4 T细胞的百分比降低(1.43%±0.96%.1.68%±0.16%,0.92%±0.21% vs 2.61%±1.27%,P＜0.01),共培养上清中IFN-γ的水平增高(578±47 mg/L,496±92 mg/L,784±97 mg/L vs 342±34 meg/L,P＜0.05),IL-4的水平降低(187±52 mg/L,169±38 mg/L,89±37 mg/L vs 226±48 mg/L,P＜0.05),以HBsAg IFN-γ组最为明显.结论:经HBsAg负载的DCs可以改善患者体内因DCs功能低下而引起的Th1细胞分化不足的状态.     

慢性乙型肝炎患者外周血CD8~+T细胞CD95表达的研究

目的检测慢性乙型肝炎患者外周血CD8+T细胞CD95分子的表达水平,了解CD95分子表达与炎症进展的关系。方法采集37例慢性乙型肝炎患者及健康对照者外周血,分离外周血单个核细胞(PBMCs),用流式细胞术检测外周血CD8+T细胞CD95分子的表达情况。结果高病毒载量组和低病毒载量组CD8+T细胞CD95的表达均显著高于健康对照组(P0.05);CD95的表达及CD8+T细胞频率之间存在负相关关系(r=-0.3486)。结论慢性乙型肝炎患者CD95的高表达可能促进了CD8+T细胞的凋亡,使CD8+T细胞的抗病毒作用减弱,可能与HBV持续感染、慢性乙型肝炎的形成机制有关。     

儿童慢性乙型肝炎患者外周血树突状细胞亚群的变化和临床特点

目的观察慢性乙型肝炎患儿外周血树突状细胞(DC)亚群的变化,探讨DC亚群在乙型肝炎慢性化中的意义.方法慢性乙型肝炎患儿组26例,HBV携带儿童组16例,健康儿童25例,利用流式细胞技术对患者外周血髓样树突状细胞(mDC)和浆细胞样树突状细胞(pDC)的百分比和绝对值进行检测,并用CPG2216刺激外周血单个核细胞(PBMC),检测pDC的α干扰素产生能力.结果与健康对照组比较,慢性乙型肝炎患儿组血mDC和pDC的百分比和数量均显著下降(P＜0.05),PBMC经CpG2216刺激后产生干扰素的能力较健康对照组也有显著下降(P＜0.05),而HBV携带儿童仅pDC百分比与健康儿童比较有明显下降(P＜0.05).相关性分析表明,HBV携带儿童pDC绝对值与HBV DNA水平之间存在正相关(r=0.5464,P=0.0351),而慢型性乙型肝炎患儿mDC绝对值与HBV DNA水平存在负相关(r=-0.4554,P=0.0194).结论慢性乙型肝炎患儿外周血DC亚群下降的频率和数量导致其功能损伤,可能与HBV持续感染的致病机制有关.     

慢性乙型肝炎患者外周血树突状细胞表型及抗原提呈能力与HBV载量的关系

目的探讨慢性乙型肝炎(CHB)患者外周血树突状细胞(DCs)表型及抗原提呈能力与HBV载量的关系.方法采集23例CHB患者和8例健康人的抗凝外周静脉血,分离外周血单个核细胞(PBMCs),在重组人白细胞介素4和重组人粒细胞-巨噬细胞集落刺激因子的作用下培养使DCs增殖、成熟,以间接免疫荧光流式细胞技术分别检测DCs表面CD80、CD86、HLA-DR及ICAM-1的表达;将培养成熟的DCs与HBsAg共同孵育,用丝裂霉素C处理后再与自体PBMCs共同培养,在培养结束前12 h加入^3H-TDR,收集细胞,以β液闪计数仪测定cpm值;同期用定量聚合酶链反应技术测定CHB患者外周血HBV载量.结果患者DCs表面CD86、HLA-DR和ICAM-1的表达水平及DCs的抗原提呈能力均显著低于健康对照组;CD80、CD86、HLA-DR及ICAM-1的表达与HBV载量呈显著负相关(分别为P＜0.01,P＜0.01,P＜0.001和P＜0.001);DCs的抗原提呈能力也与HBV载量呈显著负相关(为P＜0.001).结论CHB患者外周血DCs的成熟和功能存在障碍,DCs的抗原提呈能力与血液中HBV的载量密切相关,并可能对HBV的清除产生重要的影响.     

慢性乙肝病毒感染者外周血树突状细胞亚群与血清HBVDNA的关系研究

目的 探讨慢性乙肝病毒感染者外周血树突状细胞(DCs)亚群[髓系DC(MDC)和浆细胞DC(PDC)]的变化与血清HBVDNA的关系.方法 采集健康人和慢性乙肝病毒感染者外周静脉抗凝全血,利用荧光抗体标记和流式细胞仪检测外周血树突状细胞亚群(MDC的特异性标记为Lineage-HLA-DR+CD11c+,PDC的特异性标记为Lineage-HLA-DR+CD123+);固相放免法定量检测血清乙型肝炎病毒标志.结果 慢性乙肝病毒感染者外周血PDC相对平均百分率低于健康对照者,但缺少统计学意义,MDC相对数量在两者间没有差别;血清HBeAg阳性慢性乙型肝炎患者外周血PDC相对数量显著高(P<0.01)于HBeAg阴性患者和健康人(P<0.05),而在上述3组中MDC相对数量无差异(P=0.194).结论 HBV感染可导致患者的免疫功能改变,使机体产生免疫耐受或DC的凋亡;慢性乙型肝炎患者外周血PDC增加与HBeAg血清转换有关,提示PDC可能有抑制病毒的作用.     

青少年高血压的研究进展

随着人们生活和行为方式的改变,高血压发病明显呈年轻化趋势。在青少年时期识别高血压病高危人群有助于早期进行有效干预和治疗,降低未来高血压的发生率及其严重性。现试从青少年高血压的诊断、发病因素、特点、治疗策略等方面的研究进展作一综述。     

Morbidity in cardiovascular diseases in immigrants in Sweden

INTRODUCTION: Although immigration to Sweden has increased in the last few decades, the incidence rates of cardiovascular disease and coronary heart disease in immigrants are unknown. The aim of the present study is to estimate whether place of birth affects the incidence rates of cardiovascular disease and coronary heart disease. MATERIAL AND METHODS: The study was designed as a follow-up study on morbidity in cardiovascular disease and coronary heart disease between 1 January 1997 and 31 December 1998, including three and a half million persons with age range 35-64 years, of whom 550 000 were born abroad, from the database MigMed consisting of the whole Swedish population. Incidence rates and relative risks were estimated by indirect standardization and a proportional hazard model. RESULTS: The age-adjusted risk of coronary heart disease was higher in most foreign-born groups than in Swedes. For example, in nine of 12 male groups, the relative risks varied between 1.1 and 2.2, and in seven of 12 female groups, the relative risks varied between 1.4 and 2.5. When also adjusting for level of education and employment status, the risks were still high, but on a lower level. CONCLUSIONS: Foreign-born people possess an over-risk of cardiovascular or coronary heart disease(CVD/CHD) compared with Swedish-born persons, also when level of education and employment status are taken into account.     

Pulmonary sporotrichosis in Oklahoma in susceptibilities in vitro

Six cases of pulmonary sporotichosis were observed in 2 institutions in Oklahoma City, Okla. Three of the patients were treated with iodides with or without surgery. Although one patient required a second course of iodides, the patients have remained well after at least 34 months of follow-up. Three patients treated with amphotericin B, single course as well as multiple courses, and other antifungal agents (hydroxystilbamidine and miconazole) have all relapsed. These cases and a reviewed of more than 40 cases of pulmonary sporotrichosis susceptibilities of Sporothrix schenckii that we observed in vitro suggest that amphotericin B is not an effective agent for the treatment of pulmonary sporotrichosis. It is our opinion that the treatment of choice for pulmonary sporotrichosis is a supersaturated solution of potassium iodide. If the patient is allergic to the medication or fails to respond, then a combination of amphotericin B plus flucytosine may be tried.     

C·肉毒杀鼠素(冻干剂)应用研究

目的为研究C·肉毒杀鼠索对杀灭达乌尔黄鼠(简称黄鼠)的大面积应用情况和对家畜、家禽的毒害作用,进行了C·肉毒杀鼠素的应用研究.方法大面积投毒采用ES-2药饵撒播机[1],间隔约80m进行条投.羊、鸡采用直接灌胃.结果大面积应用的灭鼠率为83.72%.对羊、鸡最高剂量分别为500万MLD、150万MLD,均未出现中毒现象.结论 C·肉毒杀鼠素是较为理想的草原大面积杀灭黄鼠的理想、首选药物.     

Differences in outcome in primary angioplasty in women.

BACKGROUND: In acute myocardial infarction (AMI), primary percutaneous transluminal coronary angioplasty (PTCA) has proved to be the best therapeutic approach. Several factors have been associated with worse outcome in AMI in females. Are there differences in outcome in women undergoing PTCA for AMI? AIM: To evaluate gender influence on clinical outcome and in-hospital mortality in patients with AMI who undergo primary percutaneous interventions. METHODS: We studied 245 consecutive patients (72 women, 29.4 %), who underwent primary PTCA between January 2000 and December 2001. The following parameters were analyzed: risk factors for coronary artery disease including hypertension, diabetes, smoking, hypercholesterolemia and family history, previous AMI, PTCA or angina, pain-to-balloon time, extent of coronary disease and outcome. RESULTS: Female patients were older (67.9+/-11.6 vs. 59.6+/-13; p < 0.001) with a higher prevalence of hypertension (65.3 % vs. 47.4 %; p < 0.05) and angina (29.0 % vs. 16.0 %; p < 0.05) and lower prevalence of smoking (27.8 % vs. 54.3 %; p < 0.001). Pain-to-balloon time was longer in women (6.8+/-4.1 vs. 5.4+/-3.7 hours; p < 0.05). Extent of coronary disease was similar in both groups. Glycoprotein IIb/IIIa inhibitors were used in 84.7 % of women and 90.8 % of men. The frequency of hemorrhagic complications (5.6 % vs. 5.2 %) and arrhythmias (15.3 % vs. 10.4%) and in-hospital mortality (9.7 6.4 %) were higher in females, although without statistical significance (p = NS). Hospitalization time was similar in both groups. CONCLUSIONS: Despite the growing awareness of a gender bias in therapeutic approaches to AMI, there are still some differences in outcome, with a trend towards higher mortality rates in women. Older age and longer pain-to-balloon time could account for this.