不同人群细胞因子基因多态性分布的比较

目的研究上海地区部分汉族健康人群的细胞因子基因调节区或启动子区的多态性分布情况.方法随机选取上海地区60名汉族健康成人为研究对象,分别取外周静脉血2 mL,加入400 μL 5%EDTA抗凝,抽提DNA;采用序列特异性引物聚合酶链反应(PCR-SSP)法,对肿瘤坏死因子-α(TNF-α)、转化生长因子-β 1(TGF-β 1)、白细胞介素(IL)、干扰素-γ(IFN-γ)等细胞因子进行基因调节区或启动子区多态性分析.本组检测结果与采用同种方法测定的两组人群数据(文献报道)进行比较.结果三组人群IL-10的基因多态性分布差异不显著;TNF-α、TGF-β 1、IL-6和IFN-γ的基因多态性分布,三组之间差异非常明显.结论不同人群的细胞因子基因多态性分布存在差异.     

细胞因子基因型多态性与HPV引起的子宫颈损伤相关性研究

目的:探讨肿瘤坏死因子α(TNF-α)、转化生长因子β1(TGF-β1)、白细胞介素10(IL-10)、IL-18和干扰素-γ(IFN-γ)基因型多态性与人乳头瘤病毒(HPV)阳性的子宫颈损伤相关性.方法:选择本院就诊的42例子宫颈癌患者,另选择87名HPV阴性、阴道镜检查正常的妇女作为对照,检测两组TNF-α、TGF-β、IL-10、IL-18和IFN-γ的基因型多态性.结果:TNF-α、IL-10、IL-18和TGF-β表型的表达在HPV阳性与HPV阴性妇女中差异均无统计学意义(P>0.05).但IFN-γ在中度和高表达者比率HPV阳性患者明显增加(P<0.01).结论:利用细胞因子基因多态性可能有利于为HPV子宫颈癌前病变的影响进一步提供相关信息.     

细胞因子在实验性肝纤维化中动态变化及其意义

目的:探讨细胞因子TGF-β1、IL-4和IFN-γ在肝纤维化形成中变化及意义。方法:采用二硝基亚硝胺(DMN)诱导大鼠肝纤维化模型,细胞生物法测定血清转化生长因子β1(TGF-β1),双抗夹心ELISA法测定血清白细胞介素4(IL-4),肿瘤坏死因子α(TNF-α)和γ-干扰素(IFN-γ),同时进行肝组织学检查和羟脯氨酸(Hyp)测定。结果:TGF-β1、IL-4和IFN-γ在肝纤维化中均是逐步升高,TNF-γ在肝纤维化中前期升高,后期降低;TGF-β1、IL-4和IFN-γ与肝纤维化组织Hyp的相关性较好,r分别为0.71,0.52,和0.62。结论:TGF-β1、IL-4、IFN-γ、TNF-α参与肝纤维化发生的细胞因子网络的调节。     

致敏受者细胞因子基因多态性与肾移植急性排斥反应的关系

目的研究致敏受者细胞因子基因多态性与肾移植急性排斥反应的关系。方法应用序列特异性引物聚合酶链反应(PCR-SSP)对97例术前群体反应性抗体阳性肾移植受者的肿瘤坏死因子-α(TNF-α)、白介素-10(IL-10)、转化生长因子-β1(TGF-β1)、白介素-6(IL-6)、干扰素-γ(IFN-γ)的基因型进行测定,并探讨细胞因子基因型对急性排斥反应发生率的影响。结果97例致敏肾移植受者术后3个月内共有23例发生了急性排斥反应,其中TNF-α和IL-10高产量基因型组受者的急性排斥反应发生率分别高达51.9%和55.5%,显著高于相应的低产量基因型受者的12.9%和13.3%(P<0.01)。联合TNF-α和IL-10发现,高TNF-α产量基因型和高、中IL-10基因型受者的急性排斥反应发生率为62.5%,显著高于两者均为低产量基因型者的8.5%(P<0.01)。未发现TGF-β1、IL-6、IFN-γ细胞因子基因多态性与急性排斥反应的关系。结论TNF-α和IL-10基因多态性对致敏受者肾移植术后的急性排斥反应发生率有明显影响,测定TNF-α和IL-10基因型有利于制订合理的免疫抑制方案。     

贵州汉族人群肿瘤坏死因子-α基因启动子区的单核苷酸多态性

目的: 了解贵州汉族人群肿瘤坏死因子-α (tumor necrosis factor alpha, TNF-α)基因启动子区的基因多态性.方法: 采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP) 和序列特异性引物-PCR(SSP-PCR) 方法,对贵州汉族人群中TNF-α基因启动子区已知的5个多态性位点进行基因分型检测,并用统计学方法分析各位点基因型、等位基因频率及其组间差异.结果: 贵州汉族人群中TNF-α基因启动子区5个位点的等位基因频率分别为-238(G,96%;A,4.0%)、-308(G,92%;A,8%)、-857(C,65.0%;T,35.0%)、-863(C,80.0%;A,20.0%)、-1 031(T,48.0%;C,52.0%).结论: 贵州汉族人群中TNF-α基因启动子区单核苷酸多态性具有独特的特征,这为TNF-α基因多态性与疾病的相关性研究提供了基础资料.     

雾化吸入γ-干扰素对免疫低下大鼠某些细胞因子的影响

目的：研究雾化吸入γ-干扰素(IFN-γ)对免疫低下大鼠细胞因子的影响。方法：醋酸考的松皮下注射复制大鼠免疫低下模型,检测气管内注射白色念珠菌并雾化吸入IFN-γ l、3、7d的免疫低下大鼠和对照组大鼠肺泡巨噬细胞(AM)培养上清中TNF-α活性和IL-1β、IL-6水平,支气管肺泡灌洗液(BALF)IFN-γ、TNF-α活性,肺组织IFN-γ、TNF-α、IL-1β、IL-6表达,以及血清IFN-γ活性和IL-1β、IL-6水平。结果：吸入IFN-γ组大鼠AM培养上清中TNF-α的活性和IL-1β、IL-6水平显著高于对照组大鼠。吸入IFN-γ组大鼠BALF中IFN-γ、TNF-α活性高于对照组大鼠(d7 TNF-α除外)。吸入IFN-γ组灌菌后d7肺组织IFN-γ和IL-1β的表达高于对照组,TNF-α表达低于对照组,IL-6表达二组无显著差异。吸入IFN-γ组大鼠血清IFN-γ活性和IL-1β、IL-6水平,除d3 IL-1β外,与免疫低下大鼠组相比无显著差异。结论 雾化吸入IFN-γ可明显提高肺内IFN-γ、TNF-α、IL-1β、IL-6水平,但对相应血清细胞因子无明显影响。     

慢性再生障碍性贫血患者的细胞因子水平与中医证型的相关性研究

目的研究慢性再生障碍性贫血(AA)患者的细胞因子水平与中医证型的相关性。方法采用双抗体夹心ELISA法检测34例慢性AA患者的外周血γ干扰素(IFN-γ)、肿瘤坏死因子(IFN-γ)、白细胞介素-2(IL-2)、白细胞介素-3(IL-3)、白细胞介素-10(IL-10)和转化生长因子β(TGF-β)水平。结果 AA患者肾阴虚型和肾阳虚型与细胞分子的关系分析,IL-2在肾阳虚组中明显升高,在肾阴虚证中明显降低(P=0.008)。中医证型与观察指标分层讨论中显示,肾阳虚型和肾阴虚型在细胞因子的变化上有明显不同,这可能进一步说明包括IFN-γ、TNF-α、IL-2、IL-3、IL-10和TGF-β在内的多种细因子与中医辨证的内在关系,虽然不是所有的一一对等,但具有明显的统计学差异。结论中医证型与IFN-γ、TNF-α、IL-2、IL-3、IL-10和TGF-β存在明显相关性,IFN-γ、TNF-α、IL-2、IL-3、IL-10和TGF-β可能作为中医辨证的客观参考依据。     

溃疡性结肠炎虚实证候血清细胞因子表达的差异性研究

目的通过观察溃疡性结肠炎(ulcerative colitis,UC)大肠湿热证、脾气虚弱证、脾虚湿热证三种虚实证候患者血清中细胞因子TNF-α、IFN-γ、TGF-β1、IL-17A表达的差异,分析细胞因子表达与UC证候的相关性。方法分组随机纳入UC患者31例,分为大肠湿热证组、脾气虚弱证组、脾虚湿热证组,纳入10例健康志愿者作为对照组,采用ELISA法检测各组血清中TNF-α、IFN-γ、TGF-β1、IL-17A含量。UC组和对照组之间比较采用t检验,组间比较采用方差分析。结果 UC患者血清IFN-γ、TGF-β1浓度显著高于对照组(P0.01);大肠湿热证、脾气虚弱证组患者血清IFN-γ浓度均显著高于对照组(P0.01);大肠湿热证、脾气虚弱证、脾虚湿热证组患者血清TGF-β1浓度均显著高于对照组(P0.01);三种虚实证候组间比较,脾气虚弱证组患者血清TNF-α浓度显著高于大肠湿热证组(P0.05)和脾虚湿热证组(P0.01),大肠湿热证组患者血清IFN-γ浓度显著高于脾虚湿热证组(P0.01),脾气虚弱证组患者血清IFN-γ浓度显著高于脾虚湿热证组(P0.05);各组间血清IL-17A浓度比较均未见显著性差异(P0.05)。结论 UC患者血清IFN-γ、TGF-β1浓度升高可能与UC发病相关,UC患者血清TNF-α、IFN-γ的含量差异可为虚实证候的辨识提供客观实验依据。     

三种抗纤维化细胞因子对HepG2细胞转化生长因子-β1基因启动子转录活性的影响

目的:观察抗肝纤维化细胞因子白介素10(IL-10)、肝细胞生长因子(HGF)和干扰素γ(IFN-γ)对含不同-509C＞T基因型的转化生长因子β1(TGF-β1)基因上游启动子转录活性的影响,探讨细胞因子可能的抗肝纤维化机制.方法:以TGF-β1基因上游5'侧翼区启动子(-1328 ～ 812)含有-509C＞T 位点的片段作为靶序列,将其与氯霉素乙酰基转移酶(CAT)报告基因组成重组体phTGF2.14C、phTGF2.14T,脂质体法转染至人肝癌细胞系HepG2,应用IL-10(4 ng/ml)、HGF(10 ng/ml)和IFN-γ(20 ng/ml)作用于转染后HepG2细胞,ELISA法测定报告基因的表达.结果:转染phTGF2.14C的HepG2细胞报告基因活性明显高于转染phTGF2.14T者(P＜0.01).IFN-γ对转染phTGF2.14C、phTGF2.14T的HepG2细胞TGF-β1启动子转录活性均具有明显的抑制作用(P＜0.05),HGF对转染phTGF2.14C HepG2细胞的TGF-β1启动子转录活性具有促进作用(P＜0.05),IL-10对两种情况下HepG2细胞TGF-β1启动子转录活性调控作用均不显著.结论:C等位基因可明显增强TGF-β1基因启动子转录活性;IFN-γ可能通过抑制TGF-β1基因启动子转录活性抑制肝纤维化,而HGF、IL-10的抗纤维化作用可能与此途径无关.     

宁夏回、汉族群体TGF-β1启动子区基因多态性研究

目的探讨转化生长因子β1(Transforming Growth Factor-β1,TGF-β1)基因启动子区SNP-800(G/A)和SNP-509(C/T)多态性在宁夏回、汉族人群中的分布特征。方法应用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)技术对210例回族和308例汉族人群转化生长因子β1启动子区(-800位点、-509位点)两个多态性位点进行检测。结果宁夏回族群体TGF-β1基因两个多态性位点各基因型频率及等位基因频率分别为:-800(GG:97.62%;GA:2.38%;AA:0%;G:98.81%;A:1.19%),-509(CC:21.90%;CT:56.67%;TT:21.43%;C:50.24%;T:49.76%);宁夏汉族群体TGF-β1基因两个多态性位点各基因型频率及等位基因频率分别为:-800(GG:99.03%;GA:0.97%;AA:0%;G:99.52%;A:0.48%);-509(CC:27.27%;CT:50.97%;TT:21.75%;C:52.76%;T:47.24%),两组间差异均无统计学意义(P>0.05)。按性别分组,两个多态性位点的基因型频率及等位基因频率差异亦无统计学意义(P>0.05)。结论 TGF-β1基因启动子区-800、-509两个多态性位点在宁夏回、汉族人群及不同性别间差异无统计学意义。     

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.