腺病毒介导的P~(53)基因对涎腺腺样囊性癌细胞系生长的影响

目的研究P53基因对涎腺腺样囊性癌细胞系生长的影响。方法以人涎腺腺样囊性癌细胞系SACC-83、SACC-LM、ACC-2、ACC-M和NACC为实验对象,将载有人野生型P53cDNA的重组腺病毒(Ad5CMV-P53)感染涎腺腺样囊性癌细胞系,观察Ad5CMV-P53对涎腺腺样囊性癌细胞生长的影响。结果5种涎腺腺样囊性癌细胞系转染Ad5CMV-P53病毒后,均可以诱导其发生凋亡,使细胞系生长受到明显的抑制。结论Ad5CMV-P53介导的野生型P53基因,可能是一种有效的治疗涎腺腺样囊性癌的辅助手段。     

埃克替尼通过p38-MAPK信号通路对涎腺腺样囊性癌ACC-M细胞的凋亡诱导作用

目的：探讨埃克替尼对人涎腺腺样囊性癌细胞ACC-M凋亡的影响,阐明埃克替尼对涎腺腺样囊性癌的治疗作用机制。方法：ACC-M细胞随机分为对照组,2、4、8 μmo1/L^-1埃克替尼组,促分裂原活化蛋白激酶（APK）抑制剂SB203580(20 μmol/L^-1)组,SB203580(20 μmol/L^-1 )+埃克替尼(4 μmol/L^-1 )组。4 h后收
集细胞,采用MTT法检测各组ACC-M细胞的生长抑制率,用caspase-3活力检测试剂盒检测ACC-M细胞的凋亡情况（ 即caspase-3活力）,采用
Westernblotting法检测p-p38-MAPK蛋白的表达水平。结果：与对照组比较,埃克替尼组ACC-M细胞生长抑制率明显升高(P<0.05),caspase-3活力显著增强（P<0.05）,
p-p38-MAPK蛋白表达水平增加（P<0.05）。与4 μmol/L^-1 埃克替尼组比较,SB203580+埃克替尼组p-p38-MAPK蛋白表达水平明显降低（P<0.05）,caspase-3活力显著降低（P<0.05）。结论：埃克替尼可通过上调p-p38-MAPK信号的表达诱导ACC-M细胞凋亡。     

涎腺腺样囊性癌免疫组化表达的临床意义

目的观察涎腺腺样囊性癌的免疫组化表达特点,探讨CD117表达对腺样囊性癌预后的意义。方法采用组织芯片和免疫组化方法,观察41例涎腺腺样囊性癌、19例涎腺基底细胞腺瘤及31例涎腺多形性腺瘤平滑肌特异性肌动蛋白(SMA)、p63基因蛋白(p63)、S-100蛋白(S-100 Protein)、细胞角蛋白(CKpan)、上皮膜抗原(EMA)、Ki-67核抗原(Ki-67)及C-kit原癌基因(CD117)的表达,总结各抗体在涎腺腺样囊性癌、基底细胞腺瘤及多形性腺瘤中表达的强度和阳性分布模式,分析其诊断、鉴别诊断和预后意义。结果免疫组化标记SMA、p63、S-100蛋白、CKpan、EMA在涎腺腺样囊性癌、基底细胞腺瘤、多形性腺瘤中显示不同的分布特点。SMA、p63、S-100蛋白标记,腺样囊性癌肿瘤细胞巢周边及筛孔衬覆细胞呈阳性,腺管状结构外层细胞显示阳性;基底细胞腺瘤瘤细胞巢外围及腺管结构外层细胞呈阳性;多形性腺瘤阳性细胞弥漫分布或巣状分布。CKpan、EMA标记,腺样囊性癌肿瘤细胞胞质、胞膜强阳性;基底细胞腺瘤腺样结构内层细胞胞质阳性;多形性腺瘤上皮分化的肿瘤细胞标记阳性,肌上皮细胞分化的肿瘤细胞部分阳性。腺样囊性癌不同预后组Ki-67表达有差别,复发生存病例及死亡病例与无复发生存病例比较,差异有统计学意义(SNK-q检验q值分别为7.472 9和5.218 6,P<0.01)。死亡病例CD117强阳性表达与无复发生存病例比较,差异有统计学意义(P=0.039 4)。Ki-67指数高、CD117表达强阳性者预后较差。结论 SMA、p63、S-100蛋白、CKpan、EMA的不同表达对涎腺腺样囊性癌、基底细胞腺瘤、多形性腺瘤的诊断和鉴别诊断具有参考价值,Ki-67、CD117的表达与涎腺腺样囊性癌的预后相关。     

应用抑制性消减杂交技术分析涎腺腺样囊性癌转移相关基因

目的：克隆涎腺腺样囊性癌转移相关的基因。方法：应用mRNA抑制性消减杂交技术，研究涎腺腺样囊性癌高、低转移细胞系基因表达上的差异。结果：以高转移细胞ACC-M为测试子，低转移细胞ACC-2为驱赶子，获得高表达的基因序列有7个，均为已知序列同源基因。其中XAGE-1b基因为肿瘤睾丸抗原家族中的一个重要基因。结论：与ACC-2相比ACC-M中部分基因的高表达与涎腺腺样囊性癌高转移特性的获得有关，此结果为进一步探索腺样囊性癌肿瘤转移分子机理以及肿瘤转移控制和基因治疗提供了重要的依据。     

miR-26a靶向EZH2抑制涎腺腺样囊性癌ACC-M细胞的增殖活性

目的 探讨miR-26a对涎腺腺样囊性癌ACC-M细胞增殖能力的影响及其与EZH2的靶向关系. 方法 构建pcD-NATM6.2-pre-miR-26a、pmirGLO-EZH2野生型（pmirGLO-EZH2-WT）和突变型（pmirGLO-EZH2-MT）重组质粒,并采用荧光实时定量PCR（qRT-PCR）检测pre-miR-26a在ACC-M细胞中的转染效率,并与未处理组（control组）进行比较.同时采用MTT法分析miR-26a过表达对ACC-M细胞增殖活性的影响,并采用双荧光素酶报告基因系统、qRT-PCR和Western blot检测miR-26a与EZH2的靶向关系. 结果 将测序结果采用NCBI BLAST进行序列比对,结果显示pre-miR-26a、EZH2-WT和EZH2-MT重组质粒均构建成功,无碱基缺失或突变.qRT-PCR显示转染pre-miR-26a的ACC-M细胞其miR-26a的表达显著高于未处理组（P＜0.001）.MTT分析显示miR-26过表达可以明显抑制ACC-M细胞的增殖活性（P＜0.05）.此外,双荧光素酶报告基因系统（P=0.001）、qRT-PCR和Western blot均证实miR-26a能够显著下调EZH2的mRNA（P=0.001）和蛋白的表达（P=0.006）. 结论 miR-26a能够显著抑制涎腺腺样囊性癌ACC-M细胞的增殖活性,并且与EZH2之间存在良好的靶向关系.     

LAPTM4B⁃35蛋白在涎腺腺样囊性癌中的表达及临床意义

目的：检测溶酶体相关4次跨膜蛋白B（lysosome?associated protein transmembrane?4 beta,LAPTM4B）?35蛋白在正常涎腺及涎腺腺样囊性癌中的表达水平,探讨其与肿瘤患者临床病理特征的相关性。方法：收集95例涎腺腺样囊性癌和20例正常涎腺组织,以及相关临床病理资料,应用免疫组织化学法检测LAPTM4B?35在正常涎腺组织、涎腺腺样囊性癌的表达水平,卡方检验或Fisher精确检验及多因素Logistic回归分析探讨LAPTM4B?35表达水平与涎腺腺样囊性癌临床病理特征的关系。结果：LAPTM4B?35蛋白在正常涎腺组织不同细胞中呈差异性表达：在腺泡细胞中表达为阴性,在导管和闰管细胞中表达很弱,在纹状管细胞中表达中等。在涎腺腺样囊性癌组织中,48.00%的高级别涎腺腺样囊性癌LAPTM4B?35表达增高,显著高于低级别涎腺腺样囊性癌。此外,LAPTM4B?35高表达与临床分期晚亦呈显著相关。结论：LAPTM4B?35蛋白在正常涎腺组织细胞中呈差异性表达,与涎腺腺样囊性癌的组织学分级和临床分期显著相关。     

蛋白激酶CK2-α′在腺样囊性癌细胞中的表达

目的 通过检测蛋白激酶CK2-α′在涎腺腺样囊性癌中的表达分布特征,探讨蛋白激酶CK2-α′与涎腺腺样囊性癌的关系及临床病理意义.方法 应用免疫荧光技术对蛋白激酶CK2-α′在同一来源不同转移能力涎腺腺样囊性癌细胞(SACC-83)及涎腺腺样囊性癌肺转移细胞(SACC-LM)的表达进行分析.结果 蛋白激酶CK2-α′在涎腺腺样囊性癌细胞及涎腺腺样囊性癌肺转移细胞中都存在高表达,并且细胞核中的表达都高于在细胞质中的表达.结论 蛋白激酶CK2-α′的表达与涎腺腺样囊性癌的肺转移呈正相关.     

涎腺腺样囊性癌层粘连蛋白表达的研究

目的　研究层粘连蛋白在涎腺腺样囊性癌中的表达特征及其与腺样囊性癌组织分化程度的关系。方法　用超敏S P免疫组化方法检测 2 1例涎腺腺样囊性癌层粘连蛋白的表达。结果　腺样囊性癌组织分型与层粘连蛋白的表达密切相关。结论　层粘连蛋白的表达可作为腺样囊性癌组织分化的一个指标     

舌鳞癌和涎腺腺样囊性癌细胞株中MAGE-1和MAGE-3基因的表达

目的：观察肿瘤特异性抗原基因MAGE－1、MAGE－3在人舌鲜癌细胞株Tca-8113、人涎腺腺样囊性癌细胞株ACC－2和ACC－M中的表达状况，以评价这些基因作为舌鳞癌和涎腺腺样囊性癌的免疫学检测和免疫治疗、基因治疗分子标志物的可行性。方法：采取Tca-8113、ACC－2和ACC-M细胞株培养后，用Trizol试剂提取总RNA，逆转寻合成cDNA，而后应用聚合酶链反应扩增目的片段，经琼脂糖凝胶电泳确认目的片段。以GAPDH基因作为检测内对照，并与正常组织比较。结果：在人舌鳞癌细胞株Tca-8113、人涎腺腺样囊性癌细胞株ACC－2和ACC－M中，MAGE－1、MAGE－3基因均有表达。在正常组织中未见上述基因的表达。结论：MAGE－1、MAGE－3基因有可能作为舌癌和涎腺腺样囊性癌免疫学检测的分子标志物，并且具有作为免疫治疗、基因治疗特异性靶位的潜在价值。     

丁酸钠抑制涎腺腺样囊性癌细胞增殖的实验研究

目的 观察组蛋白去乙酰化酶抑制剂丁酸钠对涎腺腺样囊性癌细胞ACC-2的生长抑制情况,以及对细胞凋亡的影响.方法 采用浓度梯度为0、1.0、2.0、5.0、10.0 mmol/L丁酸钠对涎腺腺样囊性癌细胞进行干预,普通光学显微镜观察细胞形态学改变;MTT法检测细胞增殖活性;AnnexinV-FITC/PI双染流式细胞仪定量检测细胞凋亡.结果 ①0、1.0、2.0 mmol/L丁酸钠作用ACC-2细胞后,细胞无明显的形态学改变,5.0、10.0 mmol/L丁酸钠处理后,细胞变小变圆,部分细胞从瓶壁脱落,细胞增殖明显受到抑制;②1.0 mmol/L、2.0 mmol/L丁酸钠作用72 h,5.0 mmol/L、10.0 mmol/L丁酸钠作用48、72 h后,细胞生长抑制率与对照组相比P＜0.05,差别有统计学意义,且呈时间-剂量依赖性;③2.0、5.0 mmol/L丁酸钠作用ACC-2细胞72 h后平均凋亡率分别是(33.63±2.52)%和(41.10±2.48)%(P＜0.05).结论 丁酸钠呈时间-剂量依赖性影响涎腺腺样囊性癌细胞的体外增殖,并促进细胞凋亡.     

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.