肝细胞癌患者中p53基因的突变研究

为了解肝细胞癌（ＨＣＣ）中ｐ５３基因的突变情况，收集８６例ＨＣＣ手术切除术标本，采用聚合酶链反应－单链构象多态性（ＰＣＲ／ＳＳＣＰ），聚合酶链反应－限制性片段长度多态性（ＰＣＲ／ＲＦＬＰ）和ＤＮＡ序列分析，研究了ｐ５３基因的突变情况，８６例ＨＣＣ中，ｐ５３基因的突变率为３６％，其中，密码子２４９的突变率为３３．７％，１例ＳＳＣＰ和ＲＦＬＰ均提示密码子２４９存在突变者经ＤＮＡ序列分析显示密码子２４９     

食管癌组织p53抑癌基因等位基因杂合缺失及突变的研究

目的：为了探讨ｐ５３抑癌基因在原发性食管癌中的改变以及与食管癌的发生、发展可能存在的关系。方法：采用聚合酶链反应（ＰＣＲ），配合限制性片段长度多态性（ＲＦＬＰ）和单链构象多态性（ＳＳＣＰ）分析法，对４８例新疆地区原发性食管癌及正常组织中ｐ５３抑癌基因的等位基因杂合缺失（ＬＯＨ）及突变进行了检测。结果：ＲＦＬＰ分析显示ｐ５３外显子４和内含子６的ＬＯＨ分别为８／１８和４／１３。ＳＳＣＰ分析显示，ｐ５３外显子５～８的突变率为５０％（２４／４８），且突变多出现在食管癌发展的较晚阶段。结论：ｐ５３基因缺失、突变可能在食管癌发生、发展过程中起重要作用     

广西地区胃癌与P53基因突变关系的初步研究

用聚合酶链式反应－限制性片段长度多态性（ＰＣＲ－ＲＦＬＰ）技术检测８例胃癌及１例正常胃组织Ｐ５３基因有２４９密码子点突变，结果未发现异常，提示Ｐ５３基因第７外显子第２４９密码子点突变不是本地区胃癌Ｐ５３基因突变的常见类型。     

广西原发性肝癌P^53基因第249密码子点突变的研究

用聚合酶链反应－限制性片段长度多态性方法分析了原发性肝癌组织中Ｐ＾５３基因第２４９密码子的点突变，突变率为４８．４％，结果表明Ｐ＾５３基因的突变可能与环境致癌物ＡＦＢ１和ＨＢＶ有关。     

乳腺癌P53基因点突变的检测

目的：探讨Ｐ５３基因点突变检测在乳腺癌预后判断中的意义。方法：应用聚合酶链反应———单链构像多态性（ＰＣＲＳＳＣＰ）方法对３５例乳腺癌患者的Ｐ５３基因位点的点突变进行检测。结果：３例出现点突变，经ＤＮＡ序列分析，１例为第１３６密码子ＣＡＡ（Ｇｌｎ）被ＣＡＧ（Ｇｌｎ）替代，１例为第２２７密码子ＴＣＴ（Ｓｅｒ）被ＧＣＴ（Ａｌａ）替代，１例为第２７３密码子ＣＧＴ（Ａｒｇ）被ＣＡＴ（Ｈｉｓ）替代。结论：Ｐ５３基因突变可能是引起乳腺癌分化不良的因素之一，并可能预示着乳腺癌预后较差。     

白血病中P16基因突变的检测

用聚合酶链反应－单链构象多态性（ＰＣＲ－ＳＳＣＰ）、ＤＮＡ直接测序和多重ＰＣＲ法，检测了１８例慢性粒细胞白血病（ＣＭＬ），１例Ｋ５６２细胞株，９例急性粒细胞性白血病（ＡＭＬ），６例急性淋巴细胞性白血病（ＡＬＬ），２例多发性骨髓瘤（ＭＭ）患者外周血／培养细胞ＤＮＡ中Ｐ１６基因的点突变和基因缺失。用ＰＣＲ－ＳＳＣＰ共筛查出５例ＣＭＬ中Ｐ１６基因的异常，突变率为２７．８％（５／１８），对其中１例外显子２异常者经测序证实为第１５１密码子ＣＣＣ→ＣＧＣ的转换，导致Ｐｒｏ→Ａｒｇ的错义突变；并检出１例ＭＭ中Ｐ１６基因外显子３的异常；受检的ＡＬＬ，ＡＭＬ，Ｋ５６２细胞株中，未检出突变。各病例中均未检出Ｐ１６基因的缺失。本文探讨了白血病中Ｐ１６基因突变的意义。     

对非同位素PCR-SSCP检测基因突变方法的探讨

目的：建立适合本实验室条件的非同位素ＰＣＲ－ＳＳＣＰ－ＲＦＬＰ基因突变分析技术。方法：应用银染ＰＣＲ－ＳＳＣＰ技术检测石蜡包埋复发肝细胞癌（Ｈｅｐａｔｏｃｅｌｕｌａｒｃａｒｃｉｎｏｍａ，ＨＣＣ）组织中Ｐ５３基因第七外显子突变。采用系列步骤克服非特异扩增；分析影响ＳＳＣＰ诸参数的优化；并与ＲＦＬＰ结果相印证。结果：检测复发ＨＣＣ标本２２例，ＳＳＣＰ阳性率７７．２７％（１７／２２），２４９位点突变率７２．７２％（１６／２２），ＲＦＬＰ与ＳＳＣＰ分析结果符合率１００％；另有２例发生２４９位点以外的突变。结论：该实验流程可靠、高效，适合大量标本的基因突变筛查     

p53基因点突变与胃癌细胞恶性程度及临床预后的关系

为进一步明确胃粘膜癌变过程中基因变异的规律，本研究以胃癌手术标本、胃镜活检标本、体外培养细胞为研究体系，利用聚合酶链反应／限制性内切酶长度多态性（ＰＣＲ／ＲＦＬＰ）、ＰＣＲ／单链构象多态性（ＳＳＣＰ）、ＤＮＡ序列分析、免疫组化及原位杂交多项研究手段，系统研究了６０例胃癌、３０例异型增生和３３例肠上皮化生病变中基因的改变。这一结果提示ｐ５３基因异常与胃癌的发生、发展密切相关。     

人肝癌裸鼠移植瘤株p53基因突变及测序研究

目的 研究从厦门同安肝癌高发区所建的裸鼠移植瘤（ＨＨＣ，ＨＨＣ１５）是否存在ｐ５３基因突变。方法 从两移瘤提取的总ＤＮＡ用分子生物学方法检测并测序，如ＰＣＲ方法扩增ｐ５３基因部分第７外显子，地高辛标记ＤＮＡ探针斑点杂交法，ＤＮＡ限制性酶切片段长度多态分析和激光荧光ＤＮＡ测序等。结果 ＨＨＣ４细胞ＤＮＡ有ｐ５３基因第２５０密码子（Ｃ→Ａ）的突变；ＨＨＣ１５有ｐ５３基因第２４９密码子（Ｇ→Ｔ）的突变。     

肺癌组织 p53 基因突变的研究

目的：探讨肺癌发生的分子遗传学机理，了解肺癌中ｐ５３基因突变的情况。方法：采取聚合酶链反应（ＰＣＲ）及聚合酶链反应－单链构象多态性（ＰＣＲ－ＳＳＣＰ）结合银染技术，对２０例肺癌组织中ｐ５３基因外显子５～８进行了突变分析。结果：ｐ５３基因突变１２例，占总人数６０％（１２／２０），分布于外显子５～８。其中第７外显子６例，第８外显子４例，第５、６外显子各１例。鳞癌突变率５０％（３／６），腺癌突变率６０％（６／１０），小细胞肺癌突变率７５％（３／４）。结论：ｐ５３基因突变与肺癌的发生发展有关。     

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.