转染bcl-2基因对心肌细胞缺氧耐受性的影响

目的 探讨转染ｂｃｌ－２基因能否提高心肌细胞对缺氧的耐受性。方法 ①重组真核细胞表达载体。将含编码人ｂｃｌ－２ｃＤＮＡ寒带开放阅读框的ｂｃｌ－２ｃＤＮＡ重组至逆转录病毒载体－ｐＬＸＳＮ的ＥｃｏＲⅠ位点上,用脂质体包裹重度组质粒导入ＰＡ３１７细胞株,并经Ｇ４１８筛选阳性克隆;②用含高滴度重组病毒的上清感染心肌细胞,原位杂交鉴定其转染是否成功;③观察转染心肌细胞在缺氧环境下的生存率。结果 ①重组了含ｂ     

胃粘膜上皮细胞系GES—1细胞凋亡密度依赖性的研究

目的：探讨细胞密度对ＧＥＳ－１细胞凋亡的影响及其机制。方法：细胞凋亡的存在以Ｇｉｅｍｓａ染色证实，ｂｃｌ－基因的表达采用细胞原位杂交技术。结果：正常培养情况下ＧＥＳ－１细胞凋亡呈细胞密度依赖性，８ｍｇ／Ｌ的五肽胃泌素能增强ｂｃｌ－２基因的表达但不能抑制密度依赖的细胞凋亡。结论：ｂｃｌ－２基因表达可能与ＧＥＳ－１细胞凋亡的密度依赖性无关。     

粒—巨噬细胞集落刺激因子cDNA在造血祖细胞中的表达

目的；探讨逆转录病毒介导的ＧＭ－ＣＳＦｃＤＮＡ在造血母细胞中转移与表达。方法：采用电穿孔法将小鼠ＧＭ－ＣＳＦｃＤＮＡ重组逆转录病毒ｐＬＸＳＮ／ＧＭ和空载体ｐＬＸＳＮ转染病毒包装细胞ＰＡ３１７，再将含有ＧＭ－ＣＳＦ重组逆转录病毒的细胞培养上清液感染富含造血干祖细胞群体，采用ＰＣＲ和Ｓｏｕｔｈｅｒｎｂｌｏｔ分析培养细胞基因组中外源基因的整合，用Ｄｅｘｔｅｒ培养体系检测培养３周的各组细胞增殖数，结果：在     

逆转录病毒介导的TNFa基因在胶质瘤细胞系（C6）中的表达及对肿瘤细胞恶性生长的影响

利用逆转录病毒载体ＬＸＳＮ构建了含有完整编码ＴＮＦｃＤＮＡ的重组逆转录病毒质粒ｐＬＸＳＮ－ｔｎｆ，用Ｌｉｐｏｆｅｃｔａｍｉｎｅ将重组质粒导入病毒包装细胞ｐＡ３１７，经Ｇ４１８筛选培养获得抗性克隆，用ＮＩＨ３Ｔ３细胞测定病毒滴度，获得滴度为５×１０５ＣＦＵ／ｍｌ的细胞克隆。利用病毒上清液感染大鼠胶质瘤细胞系Ｃ６，得到Ｇ４１８抗性克隆细胞Ｃ６ｐＬＸＳＮ－ｔｎｆ，经ＰＣＲ检测，ＴＮＦｃＤＮＡ完整地整合在细胞基因组中。测定Ｃ６ｐＬＸＳＮ－ｔｎｆ细胞上清中ＴＮＦ的生物活性，结果显示ＴＮＦ有相对稳定的表达（４８～１８０Ｕ／ｍｌ１０６ｃｅｌｌｓ／２４ｈ）。实验还显示经ＴＮＦ基因转导的Ｃ６ｐＬＸＳＮ－ｔｎｆ细胞生长速度较之亲本肿瘤细胞Ｃ６明显下降，基因修饰后的肿瘤细胞在Ｗｉｓｔａｒ大鼠体内形成肿瘤的能力明显受到抑制。进一步用超离心法浓缩病毒对胶质瘤移植模型进行了体内治疗研究。     

癌胚抗原cDNA真核表达重组质粒的构建

目的：将人特异表达的癌胚抗原（ＣＥＡ）－ｃＤＮＡ克隆到逆转录病毒质粒载体ｐＬＸＳＮ中,构建成真核表达重组质粒ｐＬＸＳＮ－ＣＥＡ,为进一步表达表达人ＣＥＡ的动物肿瘤细胞模型作准备。方法：用内切酶ＥｃｏＲＩ酶切质粒ｐＬＸＳＮ及ｐ９１０２３Ｂ－ＣＥＡ－１７,得到线状载体ｐＬＸＳＮ及外源基因ＣＥＡ－ｃＤＮＡ,然后通过连接酶的连接反应将ＣＥＡ－ｃＤＮＡ插入到ｐＬＸＳＮ的ＥｃｏＲＩ位点,用ＥｃｏＲＩ和ＢａｍＨＩ鉴定。结果：成功地将ＣＥＡ－ｃＤＮＡ克隆到ｐＬＸＳＮ中,通过鉴定筛选获得有意义的正向重组质粒ｐＬＸＳＮ－ＣＥＡ。结论：利用基因克隆技术能将人ＣＥＡ－ｃＤＮＡ定向插入逆转录病毒质粒载体中,构建成真核表达重组质粒ｐＬＸＳＮ－ＣＥＡ,为进一步建立ＣＥＡ阳性动物肿瘤细胞模型及研究ＣＥＡ阳性肿瘤的免疫防治奠定了基础。     

反义bcl—2RNA促进HL—60细胞的凋亡

目的：分析反义ｂｃｌ－２对ＨＬ－６０细胞凋亡的作用，为进一步研究凋亡机制打基础，方法：我们用ｌｉｐｏｆｅｃｔｉｎ将插有反义ｂｃｌ－２基因片段的重组逆转病毒载体ｐＤＯＲ－ＡＢ转染ＨＬ－６０细胞，用核酸打点杂交方法检测转染效果和载体表达效果，用流式细胞仪检测ｂｃｌ－２的改变及细胞周期的变化，用光镜和电镜观察转染细胞的形态学改变，并用转染的细胞对足叶乙甙敏感性的变化间接证明反义ｂｃｌ－２的作用，结果：ｐ     

人白细胞介素-15cDNA真核表达载体的构建及其在肺癌细胞中的表达

目的观察人ＩＬ－１５ｃＤＮＡ在腺癌细胞内的表达。方法将人ＩＬ－１５ｃＤＮＡ于ＥｃｏＲＩ、ＢａｍＨＩ位点正向克隆到逆转录病毒载体ｐＬＸＳＮ,构建ｐＬ－ＩＬ－１５－ＳＮ的重组质粒。以Ｌｉｐｏｆｅｃｔｉｎ介导将该重组质粒转染到人肺鳞癌细胞（ＰＧ细胞系）和小鼠肺腺癌细胞（ＬＡ７９５细胞系）。经Ｇ４１８条件培养筛选,获得阳性细胞克隆。再经ＣＴＬＬ－２细胞增殖法测阳性细胞ＩＬ－１５的表达活性。结果获得３个ＰＧ细胞和４个ＬＡ７９５细胞阳性克隆,ＩＬ－１５活性测定显示其表达水平在２４ｈ内分别在１４２～２０１或１３８～１７８Ｕ／（ｍｌ·１０６细胞）。结论ＩＬ－１５ｃＤＮＡ转导的人和小鼠肺癌细胞可表达有生物学活性的ＩＬ－１５。     

反义寡聚脱氧核苷酸抑制HL－60细胞中bcl－2基因表达的研究

目的：研究ＨＬ－６０细胞中ｂｃｌ－２基因的表达并探索ｂｃｌ－２基因ｍＲＮＡ特异的反义寡聚脱氧核苷酸（反义ＯＤＮ）对ＨＬ－６０细胞中ｂｃｌ－２基因表达的影响．方法：采用细胞原位杂交和免疫细胞化学染色技术检测ＨＬ－６０细胞中ｂｃｌ－２基因的表达，人工合成ｂｃｌ－２ｍＲＮＡ特异性反义ＯＤＮ片段与ＨＬ－６０细胞共培养，在作用一定时间后分别测定细胞中ｂｃｌ－２ｍＲＮＡ和Ｂｃｌ－２蛋白表达的变化．结果：ｂｃｌ－２基因在ＨＬ－６０细胞中既有转录水平的高表达，又有翻译水平的高表达，人工合成的反义ＯＤＮ片段在一定浓度范围内，作用一段时间后细胞内ｂｃｌ－２ｍＲＮＡ表达无显著改变，而蛋白表达却呈现了显著下降，与浓度和作用时间呈正相关．结论：人工合成的反义ＯＤＮ片段可以抑制ＨＬ－６０细胞中ｂｃｌ－２的转录作用．在进行白血病基因治疗的实验研究中，ｂｃｌ－２基因可以作为基因治疗的一个有用的靶基因．     

霍奇金病RS／H细胞bcl—2,fas和cyclin D1蛋白表达及其相关关系

该文利用免疫组化检测了４０例霍奇金病（ＨＤ）／Ｈ细胞ｂｃｌ－２,ｒａｓ、ｃｙｃｌｉｎ Ｄ１的表达,并对其相关关系进行了研究。结果显示：ｂｃｌ－２和ｆａｓ蛋白一信号在ＲＳ／Ｈ细胞中主要呈浆膜型。在ＨＤ上亚型ＬＰ,ＮＳ,ＭＣ和ＬＤ中ｂｃｌ－２阳性细胞密度逐渐增高（Ｐ〈０．０５）,而ｆａｓ阳性细胞密度依次降低（Ｐ〈０．０５）;并呈明显负相关（ｒ＝－０．５６９０）。在４０例ＨＤ中ｃｙｃｌｉｎ Ｄ１阳性表达     

人乳腺癌MCF—7细胞凋亡过程中p53与bcl—2表达的时序性

目的：研究细胞凋亡过程中ｐ５３与ｂｃｌ－２基因表达变化的时序关系，以探索ｐ５３调控ｂｃｌ－２的可能性。方法：选择具有野生型ｐ５３的人乳腺癌ＭＣＦ－７细胞系，用５－Ｆｕ为诱导剂诱导细胞凋亡；用Ｎｏｒｔｈｅｒｎ杂交及Ｗｅｓｔｅｒｎ印迹法检测细胞凋亡过程中ｐ５３与ｂｃｌ－２表达变化的时序性。结果：ｐ５３表达增高出现在ｂｃｌ－２表达降低之前。结论：ｐ５３在时序上具有作为ｂｃｌ－２基因负调控转录因子的可能性     

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.