极低密度脂蛋白受体亚型与脂蛋白的结合效应及其摄取能力的差异

目的 探讨人极低密度脂蛋白受体（VLDLR）不同亚型与配体的结合效应及其摄取能力的差异，明确其在脂蛋白代谢和泡沫细胞形成中的作用。方法 应用基因重组技术获得分别稳定表达全长VLDLR（I型）和（Mink糖链区缺失的VLDLR（Ⅱ型）的ldlA7细胞株。核素标记VLDLR的天然配体VLDL、β-VLDL以观察配体结合能力；以VLDL温育细胞，检测细胞内脂质含量，油红O染色法观察胞内脂质堆积及泡沫细胞形成情况。结果 实验发现，稳定表达Ⅰ型VLDLR的ldl—A7细胞表面结合^125I-VLDL和^125I-β—VLDL的量与胞内甘油三酯和总胆固醇的含量均明显高于表达Ⅱ型VLDLR的ldl—A7细胞，泡沫细胞形成显著。结论Ⅰ型VLDLR与VLDL、β-VLDL的结合和摄取能力明显高于Ⅱ型VLDLR，泡沫细胞形成过程中Ⅰ型VLDLR发挥着重要作用。     

hGM-CSF受体β亚单位胞浆域在配体胞吞过程中的作用

目的　探索人粒－ 巨噬细胞集落刺激因子受体（ｈＧＭ－ Ｒ）β亚单位胞浆域在配体诱导的胞吞过程中的作用。方法　通过ＰＣＲ 扩增技术,构建β亚单位胞浆域缺失突变子（１４４１、１５８２、１７２６ 、２３５４ 和Ｓｍａ）,然后将野生型或突变型β亚单位和ＧＭ－ Ｒα亚单位ｃＤＮＡ共转染入ＣＯＳ－７ 细胞,并对转染细胞的配体胞吞进行检测。结果　以低亲和力方式与配体结合的ＣＯＳ－ ＧＭ－ Ｒα胞吞水平仅为结合配体的７％ ,而以高亲和力方式与配体结合的ＣＯＳ－ ＧＭ－ Ｒα／β胞吞水平则达３７％ ,为前者的５ 倍,这种胞吞现象可被丝氨酸／苏氨酸激酶抑制剂Ｈ－ ７ 和星形孢菌素抑制,但不被酪氨酸激酶抑制剂木黄酮和制表菌所抑制;从β亚单位胞浆域结构与胞吞关系可见,Ｃ 末端１２２ 个氨基酸缺失可使胞吞受抑,而该１２２ 个氨基酸上游的３００ 个氨基酸区域缺失则可使胞吞水平显著提高。结论　有效的胞吞需要高亲和结合力的功能受体存在,且胞吞过程不涉及酪氨酸激酶磷酸化;而β亚单位胞浆域对胞吞有正性或负性调节作用,提示这些区域含有调节胞吞过程的序列     

大鼠肝实质及非实质细胞VLDL和HDL受体的研究

作者以~(125)I-标记的人VLDL和不含apo E的HDL_3为配体,研究了大鼠肝实质细胞(PC)及非肝实质细胞(NPC)的VLDL和HDL受体的生物学特性。结果显示:VLDL受体和HDL受体均可介导肝PC和NPC结合、摄取及降解相应的脂蛋白;肝NPC上该二受体的活性分别为PC的10倍和4倍;肝NPC上VLDL受体HDL受体的解离常数分别为15.0～34.2μg/ml和10.1～17.7μg/ml,最大结合容量分别为2170～2607ng/mg和1004～2738ng/mg细胞蛋白;VLDL受体活性受EDTA抑制及胆固醇的下降调节,HDL受体不受EDTA抑制但受胆固醇的上升调节。发现apoCⅢ对VLDL受体结合功能有明显抑制作用。     

多功能极低密度脂蛋白受体

极低密度脂蛋白受体（VLDL-R）是低密度脂蛋白受体超家族的成员，主要分布于脂酸代谢活跃的组织，参与甘油三酯的代谢。然而，近年来的研究表明，VLDL-R参与多种细胞的活动，与细胞的增殖、迁移以及细胞的分化等重要细胞功能活动密切相关，故被认为是细胞内重要的多功能受体。     

HIV辅助受体结合位点配体基因的获得及合成配体体外抑制作用的研究

目的 从构建的噬菌体十肽库中筛选HIV辅助受体的配体，测序后人工合成配体并研究其体外抑制HIVgp120．抗原的活性。方法 采用噬菌体表面表达技术，构建具有一这库容量的噬菌体随机十肽库。以HIVgp120多肽做抗原，经过五轮的亲和筛选获得高亲和力的配体，测序后人工合成配体肽，以ELISA试验检测配体的抑制活性。结果 获得1个与HIV辅助受体结合位点有高亲和力的配体克隆。经核苷酸序列测定，推导出该配体的氨基酸序列并人工合成该配体多肽。结论 筛选得到的展现在噬菌体表面的配体及人工合成的配体，对HIV辅助结合位点均有较好的封闭作用。     

vMIP-Ⅱ各受体结合活性位点分析

目的 探索vMIP-Ⅱ结合趋化因子受体的活性住点以便有目的地对其进行改构。方法应用生物信息学方法对影响、vMIP-Ⅱ受体结合的氨基酸残基进行预测分析。结果vMIP-Ⅱ对不同的受体有不同结合位点。其与CC类趋化因子受体结合住点主要在核心骨架，与CXC类趋化因子受体结合位点主要在N端。结论 vMIP- 是一种极佳的趋化因子受体阻断剂类新药开发先导物。     

醉茄内酯A及其类似物与Smoothened受体的分子对接研究

目的 基于分子对接探讨醉茄内酯A及其结构简化的类似物与Smoothened受体之间的相互作用.方法 采用计算机软件,以Smoothened受体5L7I为受体模板,对醉茄内酯A及其类似物进行分子对接研究.结果 醉茄内酯A及其类似物与SMO蛋白5L7I的7次螺旋跨膜域的中心部位产生相互作用,与活性口袋的2个关键氨基酸残基结合是否牢固与生物活性密切相关.结论 醉茄内酯A及其类似物与SMO蛋白5L7I对接的结合位点信息有助于醉茄内酯类的结构简化和新型Smoothened受体抑制剂设计.     

核受体辅活化蛋白7与自噬体蛋白LC3的相互作用#br#

目的: 探索核受体辅活化蛋白7(nuclear receptor co-activator 7，NCOA7)与微管相关蛋白轻链3(microtubule-associated protein light chain 3，LC3)的相互作用，并确定其结合位点。方法: 在胃癌AGS细胞中行质谱分析，在HEK293细胞中行GST-LC3沉淀实验，分别检测两种细胞中LC3结合蛋白；采用分子克隆法构建NCOA7不同截短体和点突变体；免疫印迹法检测NCOA7的截短体ΔN462,ΔN650,ΔN650 I749/750A质粒蛋白的表达水平;GST LC3与NCOA7突变体的沉淀实验鉴定NCOA7与LC3的结合位点。结果： 质谱分析和GST-LC3沉淀实验结果表明，NCOA7是LC3结合蛋白。免疫印迹结果显示，NCOA7不同截短体和点突变体质粒蛋白均有表达。GST-LC3与NCOA7突变体的沉淀结果显示，NCOA7 N端WEDL基序和C端WEII基序是LC3结合位点。WEII基序与LC3结合受到NCOA7的651~720位氨基酸结构抑制，是一个可调节的LC3结合位点。结论： NCOA7是LC3结合蛋白，其N端WEDL和C端WEII与LC3结合。     

趋化因子受体CXCR4的结构与功能

目的 认识趋化因子受体CXCR4之结构与功能的关系。方法 分别建立野生型趋化因子受体CXCR4及CXCR2、5个CXCR4/CXCR2嵌合受体和2个CXCR4突变受体的CHO稳定表达细胞株,以配体-受体结合试验,细胞微生理监测术、体外细胞-细胞融合实验为手段观察各变异受体与重组人SDF-1β的结合能力、在受刺激后的信号转导能力,以及作为HIV-1辅助受体的能力。结果 有4个变异受体（2444a,4442,4222,CXCR4-Tr）保持了程度不同的具有辅助受体功能。结论 CXCR4以多个结构区域参与与SDF-1β的相互作用。其N端胞外区足以及具有与SDF-1β的高亲和性结合能力。第三环链对CXCR4的结合能力也具有其特定的贡献。CXCR4的信号传导不仅需要保守结构DRY盒,而且还需要贯穿整个分子的7个跨膜区域和受配体刺激后形成并维持一定的构象。CXCR4的辅助受体功能结构域与配体结合结构域间存在交叉重叠。     

结缔组织生长因子受体结合区域的预测及筛选

目的分析结缔组织生长因子(connective tissue growth factor,CTGF)二级结构,进行其受体结合区域的预测及筛选。方法利用生物信息学手段,分析CTGF二级结构、亲水性以及其它部分物理性质,预测和筛选出CTGF与受体的可能结合区域,为以后人工合成小分子多肽的CTGF拮抗剂做出初步筛选。结果通过对CTGF二级结构、相应同源蛋白对比、以及亲水性和抗原性分析结果显示,CTGF与其受体结合的位点可能位于96～102、104～112、257～272区段,它们的氨基酸序列分别是TAKDGAP、IFGGTVYRS和IRTPKISKPIKFELSG。结论初步筛选出CTGF受体结合位点,为根据其3个氨基酸序列合成CTGF拮抗剂提供了工作基础。     

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.