人拟表皮模型的建立及其屏障特性的分析

目的：建立具有正常皮肤样水渗透屏障功能的人拟表皮模型。方法：采用胰蛋白酶消化，Ｐｅｒｃｏｌ密度梯度离心方法，从健康成年女性乳房皮肤中分离纯化出活角朊细胞，按２．０×１０６的细胞密度种植于吸附有胶原蛋白的ｐｕｒｏｐｏｒｅ微孔尼龙膜上，在含有胎牛血清、表皮生长因子（ＥＧＦ）和胎牛脑垂体提取物（ＢＰＥ）的完全培养液中，浸没式培养５ｄ后转移至气－液面培养１４ｄ。结果：在上述培养条件下获得的人拟表皮具有与人皮肤相似的屏障功能，８０％拟表皮的水渗透屏障系数Ｋｐ值为（２５±６）×１０－３ｃｍ·ｈ－１。结论：该模型已可取代人皮肤用于工业毒物、药品和化妆品的经皮渗透代谢及毒性试验。     

硝酸甘油对室间隔缺损合并心力衰竭婴幼儿血流动力学影响

目的：探讨静脉滴注硝酸甘油（ｎｉｔｒｏｇｌｙｃｅｒｉｎ, ＮＴＧ）对室间隔缺损合并充血性心力衰竭婴幼儿血流动力学影响。方法：１５ 例室间隔缺损合并充血性心力衰竭婴幼儿,在连续监测心率和血压的情况下,静脉滴注ＮＴＧ０ ．４～０ ．８ μｇ·ｋｇ－ １·ｍｉｎ －１ ,利用超声心动图在用药前、用药后１ ｈ、６ ｈ 分别测定左心室射血分数（ｅｊｅｃｔｉｏｎｆｒａｃｔｉｏｎ,ＥＦ） 、短轴缩短率（ｆｒａｃｔｉｏｎａｌｓｈｏｒｔｅｎｉｎｇ,ＦＳ）、肺动脉平均压（ｐｕｌｍｏｎａｒｙ ａｒｔｅｒｙ ｍｅａｎ ｐｒｅｓｓｕｒｅ,ＰＡＭＰ） 、肺毛细血管楔压（ｐｕｌｍｏｎａｒｙｃａｐｉｌｌａｒｙ ｗｅｄｇｅ ｐｒｅｓｓｕｒｅ,ＰＣＷＰ） 和左心室壁应力（ｌｅｆｔ ｖｅｎｔｒｉｃｕｌａｒ ｗａｌｌｓｔｒｅｓｓ,ＬＶＷＳ） 等。结果：ＰＣＷＰ用药前（２．８ ±０ ．４） ｋＰａ,用药后１ ｈ 降至（１．７ ±０ ．４） ｋＰａ,６ ｈ 降至（１．６±０．４） ｋＰａ（ Ｐ＜ ０ ．０５） ;ＬＶＷＳⅠ用药前（４－３２±０－５６） Ｎ／ｃｍ２ ,用药后１ ｈ 降至（３－６５ ±０－５９） Ｎ／ｃｍ２ ,６ ｈ 降至（３－８７ ±０－６０） Ｎ／ｃｍ２（ Ｐ＜ ０．０５） ;ＬＶＷ     

Ｃａｓｐａｓｅ－３沉默对缺血缺氧大鼠骨髓间充质干细胞凋亡的影响

【目的】 探讨沉默ｃａｓｐａｓｅ－３对大鼠骨髓间充质干细胞（ＭＳＣ）在体外缺血缺氧环境下凋亡的影响。【方法】 构建靶向ｃａｓｐａｓｅ－３的ｓｈＲＮＡ重组慢病毒并转染ＭＳＣ。建立缺血缺氧模型,流式细胞术和Ｈｏｅｃｈｓｔ荧光染色法检测细胞的凋亡情况。Ｗｅｓｔｅｒｎ ｂｌｏｔ检测ｃａｓｐａｓｅ－３的蛋白表达,Ｒｅａｌ－ｔｉｍｅ ＰＣＲ检测ｃａｓｐａｓｅ－３、Ｂｃｌ－２和Ｂａｘ的ｍＲＮＡ表达。【结果】 重组慢病毒成功转染ＭＳＣ。缺血缺氧环境下,转染组的细胞凋亡率为（１３．６６ ± ０．２０）％,低于空载体组的（２１．８６ ± ０．４３）％和空白对照组的（２２．２８ ± ０．４８）％（Ｐ ＜ ０．０１）。沉默ｃａｓｐａｓｅ－３后ｃａｓｐａｓｅ－３表达下调,Ｂｃｌ－２表达上调,Ｂａｘ表达下调,Ｂｃｌ－２／Ｂａｘ比值升高（Ｐ ＜ ０．０５）。【结论】 沉默ｃａｓｐａｓｅ－３能显著提高ＭＳＣ在缺血缺氧环境下的存活能力。     

经皮穿刺球囊二尖瓣成形术(附20例临床观察)

经皮穿刺球囊二尖瓣成形术（ＰＢＭＶ）２０例，结果显示，左房平均压明显降低，术前２７７Ｋｐａ±０８１Ｋｐａ，术后１９９Ｋｐａ±０．６６Ｋｐａ，Ｐ＜００５；二尖瓣口面积明显增大，术前１３５ｃｍ２±０．１９ｃｍ２，术后２１５ｃｍ２±０．１５ｃｍ２，Ｐ＜００１，心功能明显改善，临床疗效显著。     

磷脂对大鼠离体肠管吸收葛根素的影响

目的探讨磷脂（Ｐｈｏｓｐｈｏｌｉｐｉｄｓ，Ｐ）对大鼠离体肠管吸收葛根素（Ｐｕｅｒａｒｉｎ，Ｐｕｒ）的影响。方法在改良Ｆｉｓｈｅｒ－Ｐａｒｓｏｎｓ灌流的大鼠肠管标本中，以ＨＰＬＣ法测定灌流液中Ｐｕｒ的含量。结果Ｐｕｒ的吸收峰在２５４ｎｍ处，Ｐｕｒ的标准曲线呈正相关（ｒ＝０．９９６，Ｐ＜０．０１），标准品的回收率为１００．７±４．５３％，变异系数为３．２５％，在５０ｍｉｎ时，乳化葛根素和葛根素的扩散常数分别为３×１０－７和１×１０－７ｃｍ２·ｓ－１。结论Ｐ能增加Ｐｕｒ的吸收量，提高其生物利用度     

复合培养人工皮肤的制备

复合培养人工皮肤的制备应用基础医学研究所马刚＊杨同书颜炜群张林＊侯立中用Ｄｉｓｐａｓｅ、胰蛋白酶、胶原酶Ⅰ消化分离、培养表皮角月元细胞和成纤维细胞，分别接种到胶原网架上，培养１周后改为液气界面培养。所得到的培养皮肤在组织学上于第１周时表皮呈单层；第２...     

Relationship between endothelin-1 and ischemic brain damage after subarachnoid hemorrhage and protective effect of Ginkgo biloba extract

Ｃｅｒｅｂｒａｌｖａｓｏｓｐａｓｍ（ＣＶＳ）ｉｓｏｎｅｏｆｔｈｅｍａｊｏｒｃｏｍｐｌｉｃａｔｉｏｎｓｏｆｓｕｂａｒａｃｈｎｏｉｄｈｅｍｏｒｒｈａｇｅ（ＳＡＨ）,ｗｈｉｃｈｃｏｎｔｒｉｂｕｔｅｓｔｏｔｈｅｈｉｇｈｉｎｃｉｄｅｎｃｅｏｆｍｏｒｔａｌｉｔｙａｎｄｄｉｓａｂｉｌｉｔｙｆｒｏｍｔｈｉｓｄｉｓｅａｓｅ．Ｉｎｓｐｉｔｅｏｆｅｘｔｅｎｓｉｖｅｅｘｐｅｒｉｍｅｎｔａｌａｎｄｃｌｉｎｉｃａｌｓｔｕｄｉｅｓ,ｔｈｅｐｒｅｃｉｓｅｅｔｉｏｌｏｇｙｏｆＣＶＳｒｅｍａｉｎｓｕｎｃｌｅａｒａｎｄｎｏｓａｔｉｓ…     

表皮角朊细胞的分离、培养和鉴定

表皮角朊细胞的分离、培养和鉴定马刚１，２陈世义２刘晋宇１敖艳霞＊张林２（１应用基础医学研究所２第一临床学院皮肤科）关键词Ｄｉｓｐａｓｅ角朊细胞免疫组织化学中图号Ｒ３２９．３４表皮角朊细胞培养技术正越来越广泛地应用于医学领域的科研工作中〔１〕。我们已开...     

单克隆抗体F（ab‘）2片段2种纯化方法的建立及比较

目的：建立纯化（ｍＡｂ）Ｆ（ａｂ‘）２片段的有效方法。方法：在ｐＨ５．５环境下，用ＤＴＴ激活木瓜酶，在没有ＤＴＴ存在下消化抗人肝癌ｍＡｂ ＨＡｂ１８ ＩｇＧ１，３７℃，反应８ｈ，然后分别用Ｓｅｐｈａｃｒｙｌ Ｓ－２００（１ｃｍ×６５ｃｍ）凝胶过滤柱和ＤＥＡＥ－Ｓｅｐｈａｒｏｓｅ－ＦＦ（２ｃｍ×１８ｃｍ）阴离子交换柱纯化。结果：经ＳＤＳ－ＰＡＧＥ（１００ｇ／Ｌ）电泳测定，２种方法纯化的Ｆ（ａｂ’）２     

粘附式细胞仪快速分析细胞膜脂质流动的方法

用荧光探针ＮＢＤ－Ｃ６－ＨＰＣ标记体外培养４８ｈ的小鼠肝细胞，细胞膜磷脂用５７０型粘附式细胞仪激光漂白局部细胞膜，然后测定荧光的重新分布。结果显示，荧光恢复牢为７６／，膜流动性为（１．４９±０．０５７）×１０￣（－9）ｃｍ￣２／ｓ。     

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.