乳腺癌患者BRCA1基因外显子11突变研究

目的:检测乳腺癌患者(breast and ovarian cancer susceptibility gene 1,BRCA1)基因exon11突变情况及突变位置,探讨BRCA1突变与乳腺癌的关系.方法:采取105例乳腺癌标本为实验组,30例非癌乳腺组织为对照组.应用PCR-SSCP技术和DNA直接测序法检测所有标本BRCA1基因外显子11全部序列的突变情况.结果:30例非癌乳腺组织BRCA1基因外显子11未检出突变,105例乳腺癌中有10例发生基因突变,占总例数的9.5%.10例中2例为同义突变:2430T→C,2630T→G;8例为错义突变:2532T→G,2685T→C(2例),3191C→G,3232A→G,3667A→G(2例),3876C→A .结论:BRCA1基因外显子11突变与乳腺癌的发生关系密切,对其进行检测可能对乳腺癌的患病风险评估及早期诊断具有重要意义.     

内蒙古地区妇女散发性乳腺癌BRCAl基因突变分析

目的：筛查内蒙古地区散发性乳腺癌患者中BRCAl突变位点。方法：收集新近确诊的乳腺癌患者病例及全血标本40例。以及非癌乳腺良性疾病患者全血30例作为对照,利用TIANampBloodDNA Kit试剂盒获取基因组DNA;PCR方法扩增基因组DNA的BRCAl基因第11外显子的2620～2894bp和3570～3847bp片段．所有标本利用ABl3100-DNA测序仪分析扩增的DNA片段。结果：测序的DNA片段与GeneBank中的序列进行了比对,在所扩增DNA片段中未发现碱基的突变。结论：在我们的研究中所检测的散发性乳腺癌患者的BRCAl基因第11外显子2620～2894bp和3570～3847bp区域未发现突变。     

乳腺癌BRCA1基因突变的研究

目的　乳腺癌易感基因 (BRCA1基因 )是一种抑癌基因。本研究检测BRCA1基因在乳腺癌中的突变情况 ,探讨BRCA1基因突变与乳腺癌的关系。方法　选取 89例乳腺癌患者标本作实验组 ,另取非癌乳腺组织标本 30例作对照组。每一例标本分别捣碎 ,分别用酚 -氯仿抽提法提取DNA ,每例DNA用PCR扩增BRCA1基因的 2、5、1 7、2 0等 4个外显子。分别将每例病人每个外显子的PCR扩增产物进行SSCP分析 ,对出现异常区带的PCR扩增产物进行DNA测序 ,与基因库序列比对分析其突变情况。结果　 30例非癌乳腺组织未检测出BRCA1基因突变 ,89例乳腺癌共检测出 4例突变 ,其中 2例为 5外显子的错义突变 (2 73C >G ,2 87A >T) ,2例为 1 7外显子的错义突变 (5 1 1 5T >C ,5 1 1 6A >G)。乳腺癌BRCA1的基因突变率为 4 5 %(4 / 89)。结论　BRCA1突变与乳腺癌有关系 ,检测BRCA1基因突变对于乳腺癌患病风险评估及早期诊断具有重要意义     

人食管癌组织中DNA聚合酶β基因突变的研究

目的：研究人食管癌组织中DNA聚合酶β（POLB）基因的突变情况。方法：利用逆转录聚合酶链反应（RT－PCR）、单链构象多态性（SSCP）和序列分析对30例人食管癌标本组织中DNA聚合酶β基因进行了检测。利用DNASIS、OMIGA等软件分析测序数据。结果：手术切除浸润癌组30例食管癌标本中有13例POLB发生变异,突变率为43．3％（13／30）;而癌旁对照29例正常,仅1例异常。早期原位癌组14标本中有5例POLB发生变异,突变率为35．7％（5／14）;另外,在1例食管鳞状上皮增生中也发现有POLB变异发生。在7例癌组织中存在相同58bp(177位到234位）的基因片段缺失;共有8种点突变形式：（1）375位A→G（Ile→Val),(2)454位T→C（Phe→Ser),(3)462位G→T（Tlu→终止码）,（4）466位G→A（Gly→Glu）,（5)613位A→T（Lys→Ile),(6)648位G→C（Gly→Arg),(7)660位A→G（Arg→Gly)(8)670位A→G（Glu→Gly)。结论：本研究首次在食管癌中发现POLB基因突变;该酶基因突变可能与食管癌的发生、发展相关。     

人原发性肝癌组织内抑癌基因PTEN的突变检测

目的 检测人原发性肝细胞性肝癌(HCC)组织中抑癌基因PTEN的突变。方法 采用PCR—SSCP技术和DNA测序技术检测60例HCC组织中PTEN的突变。结果 在60例配对HCC标本中，发现了共6个突变位点的存在。其中，出现在外显子内的突变有1个，即在31B号标本第4外显子中5’—侧翼下游13bP处检测到C→T的突变(89031C→T)，该突变导致了PTEN蛋白第84伉氨基酸由精氨酸变为赖氨酸。另外5个突变均位于内含子中，包括在34B号标本的第4外显子3’-侧翼下游106bP处的5碱基(AATAG)缺失突变、在9B号标本的第4外显子3’—侧翼下游67bP处检测到的一个G→T点突变(89125G→T)、在33B号标本的第6外显子3’—侧翼下游58bP处检测到的一个T→A点突变(110285T→A)、在44B号标本的第8外显子5’—侧翼上游29bP处检测到的一个C—T点突变(118833C→T)、在22B号标本的第8外显子5’—侧翼上游82bP处检测到一个A→C点突变(118780A→C)。在我们选择的SK—HEP—1、HePG2、Huh—7、ChangliVer和WRL68肝脏细胞中未检测到PTEN基因的突变。结论 肝癌组织中PTEN突变可能参与了原发性肝癌的形成。     

颅内海绵状血管畸形CCM1基因突变的分析

目的探讨颅内海绵状血管畸形（ICM）患者CCM1基因突变的情况。方法收集经手术病理证实的ICM患者25例,均为汉族,其中家族性ICM7例。以30例正常健康人作为对照组。抽取两组对象的静脉血,提取基因组DNA、PCR,扩增CCM1基因8、9、11、12、13、15、16、17、18号外显子及部分两侧相邻的内含子、DNA测序,结果与GenBank比较。结果ICM患者中有11例被检测出7个新的CCM1基因突变,突变率为44％（11／25）。其中错义突变3个：12号外显子,1160A→C（Q387P）、1172C→T（S391F）;13号外显子,1405A→C（N469H）,插入突变2个：8号外显子,704insT（K246stop）;18号外显子,2138insG（T733stop）,内含子突变1个：IVS12-4C→T,同义突变1个：17号外显子,1875C→T（F625F）。除了1875C→T的同义突变可能为基因多态性以外,其他被发现的CCM1基因突变均导致编码KRIT蛋白的变化。对照组的CCM基因未检测出异常。家族性ICM的CCM1基因突变率高于散发性ICM,两者比较差异有统计学意义（P〈0．05）。结论国人ICM患者同样存在CCM1基因的突变,导致编码的KRIT1蛋白功能改变或缺失,这与ICM发病相关,是ICM发病的遗传学基础。     

食管癌高发区人食管癌组织中DNA聚合酶β基因突变检测

目的：研究林州市食管癌高发区病例的食管癌组织中DNA聚合酶β基因(polβ)的突变情况。方法：利用PT-PCR、SSCP和序列分析对食管癌高发区50例食管癌及癌旁组织标本中polβ基因进行检测。结果：50例食管癌标本中有22例polβ发生变异,突变率为44％(22／50);而癌旁组织仅2例异常,突变率为4％(2／50)。突变形式有：58bp(177位到234位)的基因片段缺失;375位A→G(Ⅱe→Val),454位T→C(Phe→Ser),462位G→T(Glu→终止码),466位G→A(Gly→Glu),613位A→T(Lys→Ⅱe),648位G→C(Gly→Arg),660位A→G(Arg→Gly)。结论：食管癌高发区食管癌组织存在polβ基因突变,且突变率较高;该酶基因突变可能与食管癌的发生、发展相关。     

BRCAl基因突变与年轻患者乳腺癌发生的相关性分析

目的 筛查年轻乳腺癌BRCAl基因的突变位点及SNP携带情况,探讨BRCAl基因突变与年轻乳腺癌发生的关系.方法 来自我院2004年1月-2006年8月收集的乳腺癌组织共30例,其中5例有至少1个一级亲属患乳腺癌,发病年龄≤35岁.由乳腺癌组织提取基因组DNA,对BRCAl基因第2、11C、11F、11L、11I、16、20外显子的编码序列进行PCR扩增.扩增产物进行DNA直接测序证实,利用DNA Star-MagAlign软件进行序列比较.结果 BRCAl基因中共发现14个序列变异,有3个移码突变(cDNA2639、2640delTA、3343delG及3398delT)和11个点突变(cDNA 2570 C>T、cDNA2620 A>T、1473A>G、1561C>T、1594G>A、2206A>G、2227T>C、2659C>A、2806T>C、3307A>G、3375G>A),其中3个乳腺癌家族史阳性.突变率为10%(3/30).第16及20外显子未发现突变.结论 BRCAl突变主要位于第11号外显子上,乳腺癌家族史阳性的年轻乳腺癌突变率高,3个移码突变可能与年轻乳腺癌发生相关.     

食管癌高发区p53基因第五外显子突变分析

目的 :探讨高发区食管癌标本中p5 3基因的突变状况及其与危险因素的关系 .方法 :采用PCR扩增产物纯化后直接DNA序列测定技术对河南省林州市 4 0例食管癌标本p5 3基因第五外显子突变情况进行检测 .结果 :4 0例食管癌标本中 ,有 8例标本 10个位点发生突变 ,突变率为 2 0 % (8/4 0 ) .10个突变中 9个为点突变 ,1个为插入突变 .突变大多发生在G :C碱基对 ,G :C→A :T的置换最多 ,占 5 / 9,A :T→T :A的颠换占 3/ 9,G :C→C :G的颠换占 1/ 9.p5 3基因第五外显子突变与性别、年龄、食管癌家族史和吸烟无显著联系 (P >0 .0 5 ) ,但与食用酸菜有密切联系 (P <0 .0 5 ) ,其OR值为 5 .95(1.13～ 31.2 6 ) .结论 :高发区食管癌标本中 p5 3基因第五外显子突变位点较为分散 ,经常食用酸菜可引起 p5 3基因突变     

家族性乳腺癌23例患者乳腺癌易感基因-1部分序列突变分析

目的：探讨中国汉族家族性乳腺癌患者乳腺癌易感基因－1（BRCA1）的突变情况。方法：采用聚合酶链反应－单链构像多态性分析（PCR－SSCP），标记染色双脱氧末端法测序，对15个家系23例家族性乳腺癌患者进行BRCA1部分序列突变分析。结果：发现4例基因突变，突变率为17．4％（4／23例），其中1例为2228insG，致编码子711处蛋白截短，另3组（2种）为1884A→T和3232A－→G，突变引起单个氨基酸改变。结论：在中国汉族家族性乳腺癌患者中BRCA 1突变率较低，3232A→G可能为中国人的突变热点，BRCA 1突变检测在高危人群中有一定意义。     

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.     

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.