过敏性哮喘患者单核细胞来源的树突状细胞的表型及功能研究

目的 :探讨哮喘患者的外周血单核细胞 (peripheralbloodmonocyte,PBMC)来源的树突状细胞 (dendriticcell,DC)表面协同刺激分子CD1a、CD83、CD4 0、CD86的表达 ,以及其激发同种异体T淋巴细胞的能力 ,为进一步研究哮喘的发病机制及治疗提供新的思路。方法 :取哮喘患者及正常人外周血以Thomas法分离出PBMC ,然后加入相关的细胞因子及抗原培养并诱导出相应的成熟的DC细胞。用流式细胞仪 (fluorescentactivatedcellsortor,FACS)检测DC表面协同刺激分子CD1a、CD83、CD4 0、CD86的表达。成熟的DC与同种异体T淋巴细胞反应后用细胞计数板计数得出T淋巴细胞总数。结果 :哮喘患者DC的CD86表达比正常人高 (t =7.35 ,P <0 .0 0 1 ) ,但其激发同种异体T淋巴细胞的能力较对照组无明显增强。结论 :哮喘患者可表达较高水平的CD86并且有较强的激发的能力 ,表明DC在哮喘的发病中可能扮演较重要的角色     

类风湿关节炎外周血和关节液树突状细胞的膜分子表达及功能研究

目的研究类风湿关节炎（rheumatoid arthritis,RA）患者树突状细胞（dendritic cell,DC）的膜分子表达水平和功能的变化,探讨DC在RA发病机制中的作用。方法应用流式细胞术、混合淋巴细胞培养测定诱导DC细胞表面分子表达、对同种异体T细胞功能情况,诱导成熟后DC表面分子表达与临床活动及预后相关指标ESR、CRP、RF、Stoke指数的相关性。结果RA外周血来源DC表达CD83、CD209、CD80、HLA—DR较正常人外周血来源DC增高（P〈0．05）,RA关节液来源DC较RA外周血来源DC表达CD83、CD209、CD86、CD80明显增高（P〈0．05）。RA关节液组较RA和正常人外周血DC对同种异体T细胞的刺激明显增强。RA患者外周血诱导成熟DC表型CD83、CD209与stoke评分指数呈正相关。结论提示RA时关节内免疫反应可能与局部DC分化、成熟能力增强有关,后者可通过激活T细胞引起自身免疫反应,造成滑膜组织损伤。RA患者DC表面共刺激分子表达水平的增高可能与患者免疫亢进及所致的免疫损伤有关。     

细菌脂多糖对人外周血单核细胞来源的树突状细胞发育和功能的影响

目的 :探讨细菌脂多糖 (LPS)对正常成人外周血单核细胞来源的树突状细胞 (Dendritic cell,DC)表面分子表达及免疫功能的影响。方法 :分离正常人 (n=5 )外周血单个核细胞 (PBMC)以 GM-CSF、IL-4联合诱生 DC,实验组在培养的第 2天加入LPS,正常对照组不加。于第 4、8天 FACS检测 DC表面分子 CD1a、CD14、CD83、CD80、HL A-DR的表达。同期检测 DC的增殖和刺激同种异体 T淋巴细胞的增殖情况。结果 :经 LPS刺激后 ,正常人 DC早期、晚期表面分子 CD1a、CD83、CD80表达增高 ,CD14表达降低 (P<0 .0 5 )。DC数量的增加及刺激同种异体 T细胞能力明显增强。结论 :GM-SF、IL-4和 L PS联合刺激能促进正常人 DC的成熟和免疫功能的提高     

rAAV-2-HBsAg和rAAV-2-GM-CSF共感染增强人外周血单核细胞来源树突状细胞免疫刺激功能

目的探讨rAAV-2-HBsAg(表达乙肝表面抗原的重组腺相关病毒)和rAAV-2-GM-CSF(表达粒细胞/巨嗜细胞-集落刺激因子的重组腺相关病毒)共感染对人外周血单核细胞来源树突状细胞(DC)免疫刺激功能的影响.方法用rAAV-2-HBsAg和rAAV-2-GM-CSF同时感染新分离的树突状细胞;CytoTox 96 Non-Radioactive Ctoxicity Assay试剂盒检测细胞毒性T淋巴细胞(cTL)反应;混合白细胞反应(MLR)检测DC刺激同种异体淋巴细胞增殖的能力;流式细胞仪检测DC表面分子CD80、CD83、CD86和HLA-DR.结果rAAV-2-HBsAg和rAAV-2-CM-CSF共感染的DC刺激同种异体淋巴细胞增值的能力和激发的自身淋巴细胞的CTL效应增强,CD83表达水平提高.结论内源性表达的GM-CSF能促进DC成熟、增强DC的免疫刺激功能.     

TGF-β2对树突状细胞表型和功能的影响

目的: 研究转化生长因子-β2 (TGF-β2)对树突状细胞(dendritic cells, DC)表型和功能的影响. 方法: 体外培养骨髓来源的DC(BMDC)及TGF-β2诱导的DC(TGFβ2-DC). 采用双色免疫荧光化学染色或流式细胞仪(FCM)分析DC的细胞表型. 分离纯化脾脏来源的同种异体T细胞和CD4+ T细胞. 用BMDC及TGFβ2-DC刺激T细胞增殖,并用混合淋巴细胞反应(MLR)测定其能力. 在CD4+ T细胞与BMDC或TGFβ2-DC共培养5 d后,用FCM检测CD4+ T细胞表型的变化. 结果: BMDC表现为CD11c, CD86, MHC class II+和CD8а-的髓系DC. TGF-β2的诱导抑制了DC表面协同刺激分子、MHC classII的表达(P<0.01),并抑制其刺激同种异体T细胞增殖的能力. 与BMDC诱导的CD4+ T细胞相比,TGFβ2-DC诱导的CD4+ T细胞CD152(细胞毒T淋巴细胞相关分子4, CTLA-4)表达上升(P<0.01). 结论: TGF-β2的诱导促使DC表面协同刺激因子表达下降,T细胞合成CTLA-4增加,T细胞的活化和增殖受到明显抑制.     

过敏性哮喘患者树突状细胞对原始T细胞活化的影响

目的　探讨过敏性哮喘病人和正常人外周血单个核细胞 (PBMC)来源树突状细胞(DC)表达表面分子 (CD1a、CD83、CD4 0、CD86 )和细胞因子 (IL -1 2 )的差异及其对Th1和Th2型细胞因子平衡的影响。方法　分别取哮喘患者和正常人外周血经Ficoll和不连续密度梯度离心法获取贴壁细胞 ,加入细胞因子 (GM CSF和IL 4)体外培养为不成熟DC(iDC) ,给予LPS刺激使其发育为成熟DC(mDC)。另取脐血同上方法获得非贴壁细胞 ,分别加CD4单抗和CD4 5RA单抗及磁珠分离得到原始T淋巴细胞 ,分别将两组mDC和原始T细胞共培养。使用流式细胞仪 (FACS)检测树突状细胞表面共刺激分子CD1a、CD83、CD4 0、CD86的表达 ,通过ELISA法检测mDC分泌的IL -1 2及T细胞分泌的IFN -γ和IL- 4的含量。结果　 (1 )哮喘组PBMC来源DC表达CD86分子比对照组显著升高(P <0 . 0 1 )。 (2 )哮喘组DC产生IL- 1 2及其亚单位p4 0均较正常对照组显著减少 (P值均 <0 . 0 1 ) ;(3)混合淋巴细胞反应中 ,哮喘组T细胞释放Th1型细胞因子IFN -γ较对照组减少 (P <0 . 0 5 ) ,Th2型细胞因子IL -4较对照组显著增多 (P <0 . 0 1 ) ;(4)哮喘组CD86表达水平与IL 4浓度呈正相关(r=0 5 4 87,P <0 . 0 5 ) ;(5 )哮喘组IL- 1 2水平与IFN -γ水平呈正相关 (r =0 . 75 81 ,P <0 . 0     

树突状细胞表面共刺激分子在哮喘小鼠中的作用研究

目的:研究哮喘小鼠骨髓来源树突状细胞(DCs)的表型及共刺激分子在哮喘发病中的影响.方法:用卵清蛋白建立哮喘模型,利用rmGM-CSF和rmIL-4体外诱导骨髓细胞分化为DCs,流式细胞仪检测DCs表面共刺激分子的表达,混合淋巴细胞反应检测DCs刺激同种异体的T淋巴细胞增殖的能力.结果:在两种细胞因子的作用下,从骨髓中可以诱导出大量的DCs;且均表达DCs的表面标志(33D1).哮喘组DCs表达CD86分子的水平高于对照组,而CD40、CD80在两组之间没有差异;哮喘组与对照组DCs均能强烈刺激同种异体T淋巴细胞的增殖,哮喘组DCs的刺激能力明显强于对照组.结论:哮喘组DCs的抗原递呈能力增强,表明DCs在哮喘的发病中可能起着重要作用.     

依那普利对大鼠树突状细胞功能的干预效应

目的 研究动脉粥样硬化(AS)大鼠树突状细胞(DC)的功能状态及血管紧张素转化酶抑制剂(ACEI)的影响.方法 大鼠30只分组饲养后,分离外周血单个核细胞(PBMC),在含粒细胞巨噬细胞集落刺激因子(GM-CSF)、白介素-4(IL-4)培养条件下制备DC.用流式细胞仪检测DC表面共刺激分子CD86(B7-2)的表达,混合淋巴细胞反应(MLR)检测DC对同种异体T淋巴细胞的刺激能力.酶联免疫吸附法(ELISA)测定MLR上清液中细胞因子水平.结果 与正常对照组比较, AS大鼠DC表面CD86的表达明显增高,对T淋巴细胞刺激的能力增强,致炎细胞因子(IL-1β、TNF-α)分泌增多.与AS组比较,AS应用依那普利组的DC表面CD86的表达明显降低,对T淋巴细胞刺激的能力下降,致炎细胞因子(IL-1β、TNF-α)分泌减少.结论 AS大鼠DC处于激活状态,DC可能参与了AS的发病.ACEI对大鼠DC功能有明显的抑制作用,可能是其抗AS的作用机制之一.     

HBV S-ecdCD40L融合基因修饰对树突状细胞功能的影响

目的探讨HBVS—ecdCD40L融合基因修饰对树突状细胞（DC）功能的影响。方法分离健康成人外周血单个核细胞,GM—CSF、IL-4诱导培养树突状细胞,培养第5天,以脂质体介导转染,分为pcDNA3．1-S—ecdCD40L转染组、pcD—NA3．1-S转染组、pcDNA3．1转染组及PBS对照组。转染48h收集DC及上清,流式细胞仪检测DC表面CD80、CD86、HLA—DR表达水平;CCK-8法检测混合淋巴细胞反应（MLR）中DC刺激同种异体淋巴细胞增殖的能力;酶联免疫吸附试验（ELISA）检测IL-12的分泌水平。结果与对照组比较,pcDNA3．1-S—ecdCD40L转染组DC表达CD80、CD86、HLA—DR水平增加,刺激同种异体淋巴细胞增殖的能力增强,分泌IL-12水平增高。结论HBVS—ecdCD40L融合基因修饰能促进DC活化。增强DC功能,将CD40L胞外段基因与HBV抗原基因融合可能是增强乙肝疫苗免疫效果的有效方法。     

含未甲基化CpG寡脱氧核苷酸对人外周血树突状细胞抗肿瘤作用的影响

目的：研究含未甲基化CpG寡脱氧核苷酸(CpG ODN)对人外周血树突状细胞(dendritic cells，DC)分化、成熟及其抗肿瘤作用的影响。方法：分离正常人外周血DC,透射电镜观察CpG ODN刺激组和未刺激组DC的超微结构，流式细胞仪分析其表面HLA-DR、CD86的表达，MTT法检测其对同种异体混合淋巴细胞反应(mixed—lymphocyte reactor，MLR)中T细胞增殖的影响及调节T细胞杀伤肿瘤的能力。结果：与未刺激组相比，CpG ODN刺激组DC表面突起细、长且规则，粗面内质网增多，空泡减少甚至消失；同时DC表面HLA-DR、CD86的表达上调，能明显促进MLR中T细胞的增殖，增强DC调节T细胞杀伤肿瘤活性：结论：CpG ODN可诱导人外周血DC分化成熟，增强DC调节T细胞杀伤肿瘤活性。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.