24h尿液18-羟皮质醇ABC-ELISA的建立

目的建立24h尿液18-羟皮质醇亲合素-生物素酶联免疫测定法(ABC-ELISA). 方法制备18-羟皮质醇抗血清,以此建立18-羟皮质醇竞争性ABC-ELISA;同时测定部分正常人、原醛腺瘤、原醛特发性增生病人24h尿液18-羟皮质醇水平. 结果抗血清的效价为1∶30 000,交叉反应性除皮质酮为0.02%,其余均小于0.01%,批内差异为7.8%～8.2%,批间差异为9.9%～12.1%,平均回收率为101.3%,平行测定相关系数为0.9939(P＜0.01),最低可测定限为10nmol/L,正常参考值为284.7～169.2nmol/24h.原醛腺瘤组尿液18-羟皮质醇明显高于原醛增生组、原发性高血压组和正常组(P＜0.05),后三组之间无显著性差异(P＞0.05). 结论本方法具有简便、灵敏、稳定等特点,可准确反应机体24h 18-羟皮质醇的分泌情况.     

家族性腺瘤性息肉病肠粘膜、腺瘤及癌组织的基因蛋白检测与意义

研究家族性腺癌性息肉病（FAP）癌变过程中P21－H－ras、EGFR和P53的变化与意义。方法：用ABC免疫组化方法检测并比较以上三种指标在41例FAP患者的正常肠粘膜、腺瘤和癌组织标本中表达情况。结果：P21－H－ras、EGFR及P53在正常肠粘膜中阳性表达率分别为0、4．7％与0，在腺癌中为29．3％、35．4％与20．8％，癌组织中为47．4％、60．6％与52．6％，癌与腺癌的阳性率都显著高于正常肠粘膜（P＜0．05）；P21－H－ras、EGFR在癌组织中的阳性率高于腺瘤，但差别不显著（P＞0．05），P53在癌的阳性率显著高于腺癌（P＜0．05）。癌组织有2个以上指标阳性者占55．6％，3个都阳性者占27．3％；腺瘤中分别为13．2％与4．4％，差别显著（P＜0．05）。结论：提示P21－H－ras、EGFR、P53基因都参与FAP癌变，P21－H－ras基因、EGFR可能作用于早期，P53基因则作用于晚期。检测P21－H－ras、EGFR、P53对早期诊断FAP癌变有一定意义。     

内皮素抗血清抑制动脉损伤后内膜增生的实验研究

目的：通过兔颈总动脉球囊损伤模型，观察内皮素（ET）抗血清对损伤动脉内膜增生的影响，以寻找预防动脉损伤后内膜增生的新途径。方法：设立假手术组、对照组、ET抗血清组。ET抗血清组术后按0．5ml/kg/d自耳缘静脉给予ET抗血清，假手术组及照组给同等量的正常兔血清，观察术后2周各组血浆6－keto-PGF1α、TXB2水平、血管增殖指数及血管内膜厚度和管腔狭窄度。结果：1、血浆6-keto-PGF1α水平：ET抗血清组显高于对照组（P＜0．01）。2、血浆TXB2水平：ET抗血清组显低于对照组（P＜0．01）。3、损伤侧血管增殖指数：各组均显高于假手术组（P＜0．01），ET抗血清组显低于对照组（P＜0．01）。4、对照组损伤侧血管内膜厚度、截面积、管腔狭窄度均显高于假手术组（P＜0．01），而ET抗血清组上述指标均低于对照组（P＜0．01）。结论：ET抗血清抑制动脉内膜的增生。     

P53蛋白在人大肠肿瘤过度表达的研究

用鼠抗P53单克隆抗体DO—7检测25例大肠癌和35例大肠腺瘤。腺癌组按不典型增生程度分为腺瘤件重度不典型增生组、腺瘤伴中度不典型增生组、腺癌件轻度不典型增生组。各组同时计数核分裂指数（MI）。实验结果：①P53蛋白的阳性表达：腺癌组为68％．腺瘤组为34％（P＝0．01）。②70％P53阳性腺癌阳性细胞数超过40％．而腺瘤组无超过10％。③脉瘤①、②、③组P53阳性率分别为42％、38％和20％．但组间无显著差别。④MI在腺癌组高于腺瘤组（P＜0．01）．腺瘤①、②组高于③组（分别为P＜0．01,P＜0．05）。腺瘤组和腺癌组中P53阳性和阴性组与MI无统计学差别。本买验表明P53蛋白免疫组化检测,为研究大肠上皮细胞从不典型增生发展为大肠癌的肿瘤形成机理提供有意义的参考依据。     

结直肠癌缺陷基因蛋白在大肠肿瘤中的表达及临床意义

目的：探讨结直肠癌缺陷基因在大肠腺瘤－癌演序中的蛋白表达及其临床意义。方法：以结直肠癌缺陷基因蛋白胞内部分独特氨基酸链的单抗15041A为一抗，免疫组化ABC法检测21例大肠粘膜、40例伴不同程度不典型增生大肠腺瘤、21例大肠癌组织的蛋白表达情况。结果：结直怕癌缺陷基因蛋白表达率，大肠粘膜及轻度不典型增生大肠腺瘤100％，中度不典型增生腺瘤93％，明显高于重度不典型增生腺瘤（45％）及大肠瘤（33％），P＜0．05。结论：结直肠癌缺陷基因蛋白表达缺失与大肠腺瘤演变及大肠癌发展相关，免疫组化法检测结直肠癌缺陷基因蛋白发现癌前病变，研究大肠瘤演变及大肠癌发展具有重要意义。     

缺血缺氧新生大鼠脑内HO-1的表达及与CO的相关性

目的：研究缺血缺氧时新生大鼠脑内血红素氧化酶－1（HO－1）表达的变化及其与内源性一氧化碳的相关性。方法：7d龄SD仔鼠110只，随机分为3组：假手术对照组（Control,10只），缺血缺氧组（HI，50只）及缺血缺氧＋锌原卟啉组（HI＋Znpp,50只），于模型建立后0，1，4，12和24h处死，观察各组脑组织HO－1活性的变化及CO和cGMP含量。结果：（1）缺血缺氧组cGMP及CO水平在缺血缺氧后0h即升高，与对照组相比较有明显差异（P＜0．01），但与HI＋Znpp组无显著差别（P＞0．05）。（2）缺血缺氧后0h各组HO－1活性均无显著差异（P＞0．05）。（3）缺血缺氧组在1，4和12hHO－1，cGMP，CO水平与对照组及HI＋Znpp组比较均 明显升高（P＜0．01），在12h达到高峰，在24h恢复正常水平。与HI和12h均明显减低（P＜0．01），但仍高于正常组（P＜0．01）。结论：缺血缺氧对脑内HO－1诱导高表达，并与内源性一氧化碳的增高密切相关。HO－CO－cGMP系统可能在缺血缺氧脑损伤的病理生理过程中起着重要作用。     

大肠腺瘤及癌变组织中p53基因突变与bcl-2、PCNA表达关系的研究

研究p53、bcl－2及PCNA在大肠癌发生发展中的意义。应用聚合酶链反应一单链构象多态性分析检测40例腺瘤和80例腺癌中p53基因第5－8外显子突变,同时应用免疫组化SP法分别检测大肠正常粘膜,增生性息肉,腺瘤以及腺癌中p53、bcl－2、PCNA的蛋白表达。结果中17．5％（7／40）腺瘤和46．25％（37／80）腺癌可见p53基因突变;腺瘤、腺瘤癌变及腺癌中p53蛋白表达率分别为22．5％、70．6％和58．8％;正常粘膜和增生性息肉中未见p53表达,p53蛋白表达与其基因突变相关（rs=0．54）,但和临床病理因素及预后无关（P＞0．05）;bcl-2在所有正常粘膜和增生性息肉基底部上皮细胞、77．9％腺瘤、64．7％腺瘤癌变和55％腺癌可见表达,在腺瘤和腺癌中bcl－2表达差异显著（P＜0．05）,高分化腺癌bcl－2表达率明显高于差分化腺癌（P＜0．01）。应用Spearman相关分析,bcl－2与P53表达在腺瘤及腺癌组中呈负相关。PCNA表达率在大肠腺瘤一癌序列中逐渐增高,它与腺瘤p53表达和木典型增生程度、腺癌组织分化和淋巴结转移相关（P＜0．05）,但与p53、bcl－2表达及其它因素无关。p53基因改变主要涉及到良性腺瘤和腺癌的转变过程,p53基因突变是参与和影响p53蛋白表达和生物学功能的主要因素。bcl－2表达是大肠癌发生中的早期     

慢性肾衰血,尿SIL—2R的变化及意义

为探讨慢性肾衰（CRF）患者可溶性白介素－2受体（SIL－2R）变化的意义，采用ELISA法检测32例CRF患者的血、尿SIL－2R。根据Ccr分为两组：组ICcr＜10ml／min（n＝23）和组ⅡCcr＞10ml／min（n＝9）。组Ⅰ中透析达6～8周者为组Ia（n＝10），透析已＞8周者为组Ib（n＝5）。结果：1．组Ⅰ患者血SIL－2R明显高于健康对照组（P＜0．01）和组Ⅱ（P＜0．01），而尿SIL－2R无此差异。2．Ia组患者血SIL－2R透析后较透析前明显降低（P＜0．01），但组Ib无此差异。3．组Ia和Ib患者尿SIL－2R浓度在透析前后无统计学差异。4．25例CRF患者外周血CD3（％）和CD4（％）明显降低（P＜0．01），而CD8（％）趋于增高（P＜0．01）。5．血SIL－2R浓度与Ccr呈负相关，与BUN及24h尿蛋白排出量（24hUprV）呈正相关。提示：SIL－2水平可作为免疫系统活化、肌酐清除减少及有效透析的指标之一。     

粘着斑激酶在大肠癌中表达水平与侵袭，转移的相关研究

目的：研究粘着斑激酶（FAK）表达与大肠癌发生，生物学行为和预后的关系，方法：应用免疫组织化学S－P方法检测10例正常肠粘膜，16例肠腺瘤伴不典型增生及108例大肠癌手不本中FAK的表达，结果：正常肠粘膜表达均阴性，肠腺瘤伴不典型增生表达阳性率为12.50%,大肠癌表达阳性率为75．93％（82／103），其中无区域淋巴结转移者阳性率为48．84％，有区域淋巴结构转移者阳性率为93．85％（P＜0．01），FAK表达水平与大肠癌浸润兴度和Dukes分期呈正相关（均P＜0．01），FAK阳性者术后3年，5年生存率分别为49．74％，44．30％，而阴性者为100％，88．54％，二组患者术后3年，5年生存率差别均有显著性意义（均P＜0．01），结论：大肠癌由于FAK表达增加，加速恶性细胞的增殖，浸润和转移，故检测大肠活检粘膜及癌组织中FAK表达状况，有助于进一步了解大肠粘癌变倾向及大肠癌生物学行为，有利于对患者预后的判断。     

术前尿液 BLCA－4及 IL－8水平与膀胱癌临床病理特征的关系及意义

目的：探讨术前尿液BLCA－4及IL－8水平与膀胱癌病理特征的关系并确定其在临床诊断上的意义。方法：采用酶联免疫反应原理分别检测膀胱癌组患者、正常对照组和高危性膀胱癌人群中术前尿液中BLCA－4和IL－8水平,分析研究他们与膀胱癌临床病理特征的关系。结果：实验测得BLCA－4水平的Cutoff值为14．0 A／μg．protein,正常对照组尿中BLCA－4含量为2．7±5．8 A ／μg．Pro－tein;高危性膀胱癌组BLCA－4含量为9．1±18．7 A ／μg．protein,均低于实验的Cut－off值,但高危性膀胱癌组明显高于正常对照组（P＜0．01）;而膀胱癌组患者尿中BLCA－4含量为45．3±69．2 A ／μg．protein;显著高于正常对照组和高危性膀胱癌组（P＜0．001）。与正常对照组相比,术前尿液中IL－8水平在高危性膀胱癌组和膀胱癌组均显著性上升,统计学差异分别为P＜0．01, P＜0．001。膀胱癌组患者术前尿液中BLCA－4和IL－8水平均与年龄、性别、肿瘤单发还是多发,初发还是复发没有显著统计学差异（ P均＞0．05）,而与肿瘤的病理分期和分级有显著统计学差异（ P＜0．05）。结论：术前尿液BLCA－4及IL－8水平可能成为膀胱癌肿瘤诊断标志物,为临床早期诊断和病理特征判断提供依据。     

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.     

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.