共刺激分子异常介导寻常型银屑病免疫失衡的实验研究

目的通过检查外周血免疫细胞共刺激分子和细胞因子表达格局,分析寻常性银屑病患者可能发生的免疫异常。方法选择未经治疗的寻常型银屑病患者与正常对照组,抽取外周血并分离单个核细胞(peripheral bloodmonocytes,PBMC),采用RT-PCR对寻常型银屑病患者和正常对照组外周血CD86、B7H1、Foxp3、T-bet、GATA-3指标进行检测。采用流式细胞仪检查两组外周血清中IL-12、IFN-γ、IL-17、IL-2、IL-10、IL-9、IL-22、IL-6、IL-13、IL-4、IL-5、IL-1β、TNF-α含量。结果寻常型银屑病组PBMC中Foxp3和GATA3表达低于正常人群,但T-bet表达增高(P<0.05)。寻常型银屑病患者TNF-α明显高于正常对照组(P<0.05)。寻常型银屑病患者CD86相对表达量(0.3636±0.1003)明显高于对照组(0.1061±0.03883)(P<0.05);而B7H1相对表达量(0.06876±0.03145)明显低于对照组(0.3401±0.1152)(P<0.05)。结论寻常型银屑病患者Th1亚群增加、TNFa水平增高,而Th2和调节性T细胞(Regulatory T cells,Treg)亚群明显降低;寻常型银屑病患者外周血抗原递呈细胞(antigen present cells,APC)表达共刺激分子CD86增加而共抑制分子B7H1降低,提示寻常型银屑病患者体内存在的免疫功能紊乱可能是由于其APC功能异常所致。     

银屑病患者外周血单个核细胞CCR5的表达及其意义

目的研究银屑病患者外周血单个核细胞趋化因子受体5(CCR5)的表达及其与银屑病皮损面积和严重程度指数(PASI)的关系。方法分离35例中重度寻常型银屑病患者及30例健康对照者外周血单个核细胞(PBMCs),用RT-PCR法测定PBMCs培养前后CCR5 mRNA的表达水平,免疫荧光标记-流式细胞仪检测CCR5阳性细胞比率。结果治疗前银屑病患者外周血单个核细胞中CCR5 mRNA表达水平明显高于治疗后及健康对照组,并与PASI呈正相关(r=0.516,P<0.05)。结论 CCR5可能通过活化与趋化单个核细胞到银屑病皮损而参与了银屑病的发病。     

趋化因子受体CXCR3 mRNA在寻常型银屑病患者皮损及外周血单个核细胞中的表达

目的 研究手常型银屑病患者皮损及外周血单一核细胞(PBMC)中趋化因子受体CXCR3 mRNA的表达及其临床意义.方法 RT-PCR法测定20例银屑病患者皮损和10例健康正常人皮肤及其PBMC中CXCR3 mRNA的表达;直线相关性分析方法分析银屑病患者皮损及PBMC中CXCRB的表达水平与银屑病皮损面积和严重程度指数(PASI)之间的关系.结果 银屑病患者皮损及PBMC中CXCR3 mKNA表达相对含量分剐为(1.44±0.72)和(1.49±0.10),正常人皮肤及PBMC中CXCR3 mRNA相对含量为(0.51±0.10)和(0.45±0.09).经统计学分析,银屑病皮损及外周血PBMC中CXCR3的mRNA表达明显高于正常对照组,差异有显著性(P<0.05);银屑病患者CXCR.3 mRNA的表达水平与PASI呈正相关(P<0.05).结论 趋化性因子受体CXCR3可能参与了在银屑病的发病过程.     

寻常型银屑病患者外周血线粒体DNA检测及其临床意义

目的： 分析寻常型银屑病患者外周血线粒体DNA（mtDNA）水平,并探讨其在疾病中的作用机制。方法: 收集28例寻常型银屑病患者和28例健康对照者外周血,实时荧光定量PCR检测血浆mtDNA表达。分离外周血单个核细胞,实时定量PCR检测环状GMP-AMP合酶（cyclic GMP AMP synthase,cGAS）和干扰素基因刺激物（stimulator of interferon gene,STING）mRNA表达水平。外周血单个核细胞与mtDNA共培养,ELISA检测上清液白细胞介素（interleukin,IL）-23、IL-17、干扰素-γ（interferon-γ,IFN-γ）浓度。结果: 与健康对照组相比,银屑病组外周血mtDNA相对表达显著升高\[(2.34±0.81) vs (1.18±0.25),P<0.001］,且与银屑病皮损面积和严重程度指数（PASI）评分呈正相关（r=0.693,P<0.001）。与健康对照组相比,银屑病组外周血单个核细胞cGAS和STING mRNA相对表达均显著上升（P均<0.001）。与mtDNA共培养后,单个核细胞IL-23、IL-17、IFN-γ分泌显著增加（P均<0.001）,且与PASI评分均呈正相关（r分别为0.647,0.585,0.492,P均<0.01）。结论: 银屑病患者外周血mtDNA异常升高,可能通过活化免疫细胞cGAS/STING信号通路,参与银屑病疾病过程。     

SLE患者外周血单个核细胞中信号转导子及转录激活因子4mRNA的表达及其与血清IFN-γ水平之间的相关性

目的探讨系统性红斑狼疮(SLE)外周血单个核细胞(PBMC)中信号转导子及转录激活因子4(STAT4)mRNA表达及其与血清干扰素(IFN-γ)水平之间的相关性。方法用TaqMan实时荧光定量聚合酶链反应法检测60例SLE患者及30例正常对照者PBMC中STAT4mRNA表达水平,用酶联免疫吸附法检测SLE患者血清中IFN-γ的水平,并分析两者之间的相关性。结果 PBMC中STAT4mRNA表达水平在SLE患者组中要显著高于正常对照组(P<0.05),并且在SLE患者病情活动组中显著高于病情非活动组(P<0.05)。SLE患者血清中IFN-γ水平检测结果为(12.71±5.68)ng/ml,并与患者PBMC中STAT4mRNA表达呈正相关(r=0.769,P<0.05)。结论 SLE患者PBMC中STAT4mRNA的表达水平高于正常,并与病情活动性及患者血清中IFN-γ水平相关。     

寻常型银屑病患者血清S100A12水平的测定及临床意义

目的了解寻常型银屑病患者血清中S100A12水平及其临床意义。方法 ELISA法检测50例寻常型银屑病患者和50例健康对照组血清S100A12水平,并计算患者银屑病面积和严重程度指数(psoriasis area and severity index,PASI)。使用Pearson分析法分析S100A12水平与PASI相关性。结果寻常型银屑病组血清S100A12水平为(154.4±114.9)ng/mL;对照组血清S100A12水平为(72.6±62.8)ng/mL。寻常型银屑病组S100A12水平高于对照组,差异有统计学意义(P0.05)。Pearson分析结果显示寻常型银屑病组S100A12水平与PASI评分之间呈正相关性(r=0.436,P0.05)。结论银屑病患者血清S100A12水平明显高于健康人,与疾病严重程度相关,可作为判断银屑病严重程度的指标。     

寻常性银屑病患者外周血单个核细胞TLR4mRNA表达与TNF—α相关性

目的探讨寻常性银屑病患者外周血单个核细胞（PBMC）Toll样受体4（Toll-like receptor4,TLR4）的表达变化及其与血清中TNF—Q浓度的关系。方法采用荧光定量PCR技术检测寻常性银屑病患者外周血单个核细胞TLR4mRNA的表达,并采用双抗体夹心酶联免疫吸附法（ELISA）检测血清TNF—α的浓度。另取30例正常人作为对照组。结果寻常性银屑病患者外周血中PBMC的TLR4mRNA的表达较正常人对照组明显增高（P〈0．01）,TLRdmRNA表达与血清TNF-α浓度变化呈正相关（P〈0．05）。结论通过单核吞噬系统TLR4介导的,TNF-α大量产生,激活炎症级联反应可能是银屑病发病的重要病理机制之一。     

颈段食管癌患者外周血单个核细胞HLA-A和HLA-B mRNA的表达

目的检测颈段食管癌患者外周血单个核细胞(peripheral blood mononuclear cell,PBMC)HLA-A和HLA-B mR-NA的表达水平,探讨其在肿瘤免疫中的作用及其临床应用价值。方法收集32例颈段食管癌患者和45名健康体检者外周血单个核细胞,用实时荧光RT-PCR方法检测淋巴细胞HLA-A和HLA-B mRNA的表达水平。结果PBMC的HLA-A和HLA-B mRNA在颈段食管肿瘤组的相对表达率分别为0.41±0.26和0.37±0.28,在正常对照组分别为1.12±0.54,1.19±0.60,颈段食管肿瘤组HLA-A和HLA-B mRNA相对表达率低于正常对照组(P均<0.000 1)。多因素Logistic回归分析显示,HLA-A,B mRNA的表达与颈段食管癌的发生有关(P=0.01)。结论颈段食管癌患者外周血单个核细胞HLA-A和HLA-B mRNA表达水平低于正常对照组。HLA-A和HLA-B mRNA的表达水平或许可以作为一个新的肿瘤标志物,这对颈段食管癌的早期诊断和治疗提供了一个新的思路和方向。     

2型糖尿病伴周围神经病变患者外周血单个核细胞中醛糖还原酶mRNA水平测定

目的探讨外周血单个核细胞(PBMC)中醛糖还原酶(AR)mRNA水平与2型糖尿病并发周围神经病变的关系.方法用RT-PCR法检测32例2型糖尿病患者及35名正常对照者PBMC中ARmRNA水平,用肌电图检测腓神经传导速度,根据神经传导速度有无减慢将糖尿病患者分为普通糖尿病组和糖尿病神经病变组.结果2型糖尿病患者PBMC中AR mRNA的表达量(0.59±0.56)高于正常对照组[(0.30±0.1l),P=0.003];糖尿病是否伴神经病变患者之间AR mRNA的表达量无差别.结论2型糖尿病患者PBMC中AR mRNA表达水平增高与是否伴神经病变无关.     

信号转导和转录激活因子4及γ-干扰素在银屑病患者皮损中的表达及其意义

目的研究银屑病患者皮损内信号转导和转录激活因子4(STAT4)及γ-干扰素(IFN-γ)的表达及其临床意义。方法利用荧光定量聚合酶链反应法测定20例寻常型银屑病患者皮损内及17例正常对照者皮肤中STAT4 mRNA及IFN-γmRNA的定量表达,采用直线相关回归分析方法分析STAT4mRNA及IFN-γmRNA的表达与银屑病病情严重指数(PASI)的相关性,STAT4mRNA与IFN-γmRNA表达的相关性。结果银屑病皮损内STAT4mRNA及IFN-γmRNA的表达均高于正常对照者皮肤内的表达水平(P<0.001)。银屑病皮损内STAT4mRNA的表达与患者PASI呈显著的正相关(r=0.78465,P<0.0001),IFN-γmRNA的表达与患者PASI无显著相关性(r=-0.16125,P=0.4970)。银屑病皮损内STAT4mRNA的表达与IFN-γmRNA的表达呈显著的正相关(r=0.62149,P=0.0034)。结论STAT4可能通过上调IFN-γ的表达,促进银屑病的炎症进程。     

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.     

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.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.     

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.