肺癌纤维支气管镜刷落细胞中p16基因外显子2丢失的研究

目的：探讨p16基因外显子2丢失与原发性肺癌发生发展之间的相关性，以及纤维支气管镜刷落细胞中检测p16基因外显子2丢失代替手术标本检测的可行性。方法：在纤维支气管毛刷脱落细胞中采用PCR－电泳－紫外成像条带密度扫描法，以β－actin基因为内对照，检测及分析病理证实的原发性肺癌患者病灶侧与其相对正常侧p16基因外显子2丢失的情况。结果：p16基因外显子2丢失率在肺癌患者相对正常侧毛刷脱落细胞中0（0／19），在病灶侧毛刷脱落细胞中35．5％（11／31），两者比较有统计学显著差异（P＜0．01）。小细胞肺癌（SCLC）标本中丢失率为0（0／7），非小细胞肺癌（NSCLC）标本中丢失率为50％（11／22），两者比较有统计学显著差异（P＜0．05）。结论：p16基因外显子2丢失可能与NSCLC的发生发展相关。纤维支气管镜直视下病灶处毛刷脱落细胞检测p16基因外显子2丢失率可代替手术标本以协助术前诊断与治疗。     

p16基因第二外显子纯合性缺失检测对肺癌所致胸液的临床价值

目的　探讨p16基因第二外显子纯合性缺失检测对肺癌所致胸腔积液的临床价值。方法　用PCR技术检测 31例肺癌胸液及 21例结核性胸腔积液中p16基因第二外显子纯合性缺失的情况,同时送检胸液脱落细胞学作为对照分析。结果　表明所检 21例结核性胸液无p16基因第二外显子纯合性缺失, 31例肺癌胸液标本中有 12例出现第二外显子纯合性缺失,缺失率为 38 7% ( 12 /31 )。同时 16例脱落细胞学检测阳性,阳性率为 51 62% (16 /31)。在 15例脱落细胞学阴性中,有 6例存在p16基因第二外显子纯合性缺失,两者联合检测阳性率提高至 70 97% (22 /31),明显高于单独检测(P<0 05)。另外,p16基因第二外显子纯合性缺失与肿瘤细胞的组织学类型、转移、预后相关。本组研究表明鳞癌出现p16基因第二外显子缺失的机率虽高于腺癌但差异无显著性;而有淋巴结转移者明显高于无淋巴结转移者(P<0 005)。同时,在 12例p16基因纯合性缺失的患者中有 9例已发生一处以上的肺外转移,预后极差。结论　检测胸液p16基因第二外显子纯合性缺失对肺癌胸腔积液的诊断有重要价值,且特异性高;与脱落细胞学联合检测,可补充和提高其诊断阳性率,对鉴别癌性和结核性胸腔积液及判断肺癌的预后有一定的参考意义。     

原发性非小细胞肺癌中p16/MTS1基因外显子纯合性缺失的研究及临床意义

目的 检测原子性非小细胞肺癌（Non-small Cell Lung Cancer,NSCLC)手术标本p16基因（Multiple Tumor Suppressor Gene,MTS1)第一、二外显子纯合性缺失并探讨其与肿瘤细胞分化程度、组织类型、转移和预后的关系。方法 收集我院1999年1月－12月原发性支气管肺癌且手术病理证实为非小细胞肺癌32例。提取DNA,建立PCR方法分析p16基因第一、二外显子纯合性缺失,用Southern Blotting方法对p16第二外显子的PCR产物进行鉴定。结果 32例NSCLC患者中9例发生p16基因第二外显子的缺失,缺失率为28％（9／32）,p16第一外显子未发现缺失。结论 在原发性非小细胞肺癌中,p16基因第二外显子的纯合性缺失是p16基因失活的一个重要方式,且影响肺癌细胞其他生物学行为。     

CDKN2／p16基因在四种原发恶性肿瘤中存在状态的研究

目的：研究CDKN2/16基因缺失和蛋白丢失在非小细胞肺癌，食管癌、胃癌、乳腺癌4种原发恶性肿瘤中的作用。方法：采用免疫组织化学和多重PCR技术对68例（非小细胞肺癌31例，食管癌15例，胃癌13例，乳腺癌9例）原发肿瘤组织标本进行CDKN2/p16基因第一，第二外显子纯合缺失和p16蛋白表达丢失分析研究，结果：第一或（和）第二外显子总缺失率为13．2％（9／68），蛋白丢失率为44％（30／68），不同肿瘤组织标本的p16基因缺失率和蛋白丢失率差别很大。结论：CDKN2/[16基因是重要的多肿瘤抑制基因，在不同肿瘤组织中其失活方式不完全相同。     

复发性星形细胞瘤p16基因的研究

目的：结合病理学分级，研究复发星形细胞瘤的p16基因异常，探讨p16基因在星形细胞瘤复发中的作用。方法：采用复合PCR、PCR-SSCP等方法，对22例星形细胞瘤患的原、复发标本进行基因缺失及点突变的检测。结果：在全部44个标本中发现纯合性缺失14例，缺失率为31．8％(14／44)。将Ⅰ和Ⅱ、Ⅲ和Ⅳ级分别合并后相比差异有显性。发生p16基因缺失的病例，依其发生的时间先后分为三组。PCR-SSCP检测，点突变发生率为6．8％(3／44)。结论：星形细胞瘤在其恶性增殖中发生p16基因缺失，而基因缺失可能在维持其恶性表型中起重要作用。检测p16基因的缺失对判断星形细胞瘤的恶性转化有一定价值。点突变在星形细胞瘤进展中发生频率较低，可能不起重要作用。     

膀胱癌细胞p16抑癌基因失活的研究

目的 探讨膀胱癌中p16基因失活情况。方法 采用PCR、PCR—SSCP和DNA甲基化分析技术等方法对25例膀胱癌组织中p16基因进行检测。结果 25例膀胱癌标本中有6例存在p16基因纯合性缺失，缺失率为24％；无1例出现点突变；16例出现异常甲基化，检出率为69．6％(16／23)。结论 外显子2的纯合性缺失在安徽区域发生的膀胱癌中可能不是一个较频繁发生的事件；通过CpG岛异常甲基化失活可能是膀胱癌p16基因失活的主要机制。     

卵巢癌p16基因的纯合性缺失、突变及表达异常的研究

目的：研究p16基因的纯合性缺失，突变及表达异常与卵巢癌的发生是否存在相关关系。方法：采用PCR-SSCP检测卵巢癌p16基因的纯合性缺失和突变；采用RP-PCR定性及半定量分析p16基因；采用免疫组化技术检测p16蛋白。结果：32例卵巢癌中检测到1例外显子1突变，未检测到p16基因的纯合性缺失；RT-PCR分析卵巢癌中p16基因mRNA均有表达，半定量分析卵巢癌与正常卵巢组织中p16基因mRNA表达水平无差别；免疫组化检测9例卵巢癌中的p16蛋白为阴性（28.2%)。结论：卵巢癌的发生与p16基因的纯合性缺失和突变无相关关系，与p16基因异常表达有相关关系。     

γ射线诱发小鼠白血病模型中p16基因的改变

目的：研究γ射线诱发小鼠白血病模型中p16基因的失活机制。方法：应用聚合酶链反应(PcR)扩增39例γ射线诱发的白血病和对照小鼠胸腺组织中p16基因第1、第2外显子，检测等位基因纯合性缺失；用限制性内切酶-PCR法检测p16基因的甲基化发生情况；并应用PCR-单链构象多态性分析银染方法检测p16基因碱基突变的发生。结果：在白血病模型中，外显子2的缺失率为33．3％，甲基化的发生率为23．1％。结论：γ射线诱发的小鼠白血病模型发生、发展过程中伴有p16基因的改变，其失活以纯合性缺失和甲基化为主。     

急性淋巴细胞白血病中p16基因甲基化及表达

目的：探讨p16基因甲基化在急性淋巴细胞白血病发生机制中的作用。方法：采用限制性内切酶酶切结合多聚酶链反应(PCR)方法检测82例急性淋巴细胞白血病患者白血病细胞p16基因部分启动子和外显子1 CpG岛甲基化的情况，同时应用免疫组织化学染色法检测p16蛋白的表达。结果：31例急性淋巴细胞白血病患者存在p16基因的甲基化；1例p16基因纯合缺失；39名患者p16蛋白表达阴性；p16蛋白阴性表达患者的初次化疗完全缓解率和平均生存时间低于p16蛋白阳性表达者。结论：p16基因甲基化在急性淋巴细胞白血病的发生机制中有一定的作用，p16蛋白可能是评估急性淋巴细胞白血病预后的一个指标。     

FHIT和p53基因在非小细胞肺癌中的表达

目的研究脆性组氨酸三联体（FHIT）基因与p53基因在非小细胞肺癌（NSCLC）中的表达及其作用。方法病理确诊的NSCLC标本113例,免疫组化法检测FHIT和p53基因的蛋白表达,以癌旁正常组织作为对照。结果113例NSCLC中,66例（58％）FHIT蛋白表达降低,35例（31％）p53蛋白表达增多,均以鳞癌和吸烟患者多见（P〈0．01）。结论FHIT基因缺失与p53基因异常,可能在NSCLC的发生和发展中起重要作用。     

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.