人单纯疱疹病毒1型对血管平滑肌细胞脂质代谢的影响

目的：研究人单纯疱疾病毒1型（HSV－1）对血管平滑肌细胞（SMC）脂质代谢的影响，探讨HSV一且与动脉粥样硬化（AS）的关系。方法；以HSV－1感染体外培养的牛主动脉平滑肌细胞，观察细胞形态和超微结构的改变，以及细胞内脂质代谢的变化。结果：HSV－1感染后，用PCR法能检出SMC内存在HSV－1核酸片段；感染后24h和36h，感染组细胞内总胆固醇（CH）较对照组明显增高（P＜0．05），HE染色和电镜下均显示感染组细胞空泡样变。结论：HSV－1不但能在体外感染血管SMC，且能导致SMC内胆固醇积聚，可能促使SMC向SMC源性泡沫细胞转变，从而参与动脉粥样硬化的发病过程。     

内皮细胞对平滑肌细胞增殖和脂质代谢的影响

本研究证明:自兔血浆分离得到的低密度脂蛋白(LDL)具有促进家兔主动脉平滑从细胞(SMC)的体外增殖作用,LDL与融合的家兔主动脉内皮细胞(EC)作用后的条件培养液则无促SMC增殖作用;稀疏生长EC条件培养液能提高SMC的LDL受体活性,促进胞内胆固醇积聚;LDL与融合EC作用后的条件培养液能抑制SMC LDL受体活性,抑制胞内胆固醇积聚;并使与天然LDL竞争结合LDL受体的能力明显下降。上述结果提示:不同生长状态的EC对SMC增殖及脂质代谢起不同的作用。     

99mTc标记的寡核苷酸在人卵巢癌细胞中的摄取及其对目的基因表达的影响

目的研究针对增殖细胞核抗原(PCNA)mRNA的反义寡核苷酸的标记方法,标记后探针在卵巢癌细胞株HO8910中的摄取情况及其对细胞增殖和目的基因表达的影响。方法人工合成一段针对PCNAmRNA的反义寡核苷酸(ASON),以放射性核素99mTc进行标记,研究标记物的标记率,放射化学纯度,与互补正义链寡核苷酸(SON)的结合能力,不同温度条件下在卵巢癌细胞中的摄取情况,以及反义探针对细胞增殖和细胞PCNAmRNA表达的影响。结果在双功能螯和剂NHSMAG3的作用下,ASON的标记率为(60.12±3.01)%(n=5),经纯化后放射化学纯度可达95%以上;标记物在体外能稳定存在,且仍保持与互补链的结合能力;37℃和22℃条件下,细胞对ASON的摄取率高于对SON,对ASON的清除速度缓于对SON,而37℃时,细胞对ASON的摄取明显高于22℃时,清除速度则明显快于22℃。标记的ASON与细胞孵育48h后,HO8910细胞中PCNAmRNA的表达显著减少。结论在双功能螯和剂的作用下,寡核苷酸能成功地得到标记,与SON相比,反义探针能被代谢旺盛的HO8910细胞特异性摄取并抑制目的基因的表达。     

环氧合酶-2反义寡核苷酸对宫颈癌Hela细胞顺铂敏感性的影响

目的研究COX-2反义寡核苷酸(AS-ODNs)对宫颈癌Hela细胞顺铂敏感性的影响。方法体外设计、合成针对COX-2的反义寡核苷酸,并转染宫颈癌Hela细胞,应用细胞蛋白印记法(West-blot)测定转染COX-2反义寡核苷酸后Hela细胞COX-2基因蛋白的表达变化,应用四甲基偶氮唑蓝比色法检测(MTT)COX-2反义寡核苷酸转染后Hela细胞对顺铂敏感性的变化。结果转染针对COX-2的反义寡核苷酸后,在24、48和72h,Hela细胞COX-2蛋白的表达水平下降;在转染48h后,Hela细胞对顺铂的半数抑制量从(10.475±1.713)μg/ml提高到(0.194±0.021)μg/ml,Hela细胞对顺铂的敏感性增加了53.88倍。结论COX-2反义寡核苷酸能增加宫颈癌Hela细胞对顺铂的敏感性。     

端粒酶基因hTRT反义寡核苷酸对其酶活性和肿瘤细胞增殖的影响

（1）目的：探讨端粒酶基因hTRT反义寡核苷酸对肿瘤细胞增殖和端粒酶活性的影响。（2）方法：采用MTT法检测反义寡核苷酸对肿瘤细胞HL－60增殖的影响。采用TRAP－PCR法检测反义寡核苷酸处理肿瘤细胞后端粒酶活性的变化；采用Giemsa染色观察反义寡核苷酸处理后肿瘤细胞形态学变化，采用DNA凝胶电泳和三苯胺法检测反义寡核苷酸处理后肿瘤细胞DNA片段化。（3）结果：端粒酶基因hTRT反义寡核苷酸作用48h对肿瘤细胞增殖出现抑制作用。5μmol/L反义寡核苷酸处理24h后，细胞端粒酶活性下降，并且随作用时间的延长，活性不断降低，96h下降至对照组的44．4％，用5μmol/L反义寡核苷酸处理96h的HL－60细胞出现凋亡的形态学特征，处理24，48，72，96h后凋亡指数为0.053,0.127,0.257,0.315,，用5μmol/L反义寡核苷酸处理HL－60细胞96h后，电泳出现DNA梯状条带。作用24，48，72，96h细胞DNA片段化率分别为17．98％，29．34％，35．43％，41．24％；（4）结论：端粒酶hTRT基因反义寡核苷酸能抑制肿瘤细胞的增殖和端粒酶活性，并能诱导肿瘤细胞HL－60凋亡。     

IL-6反义寡核苷酸对骨吸收的抑制作用

目的 :探讨IL -6反义寡核苷酸在体外对骨吸收的抑制作用及意义。方法 :分离培养新生小鼠颅盖骨 ,并分别用IL -6硫代反义寡核苷酸 (ASON )、无义寡核苷酸 (NSO )、IL -6或ASON与IL -6联合作用颅盖骨 ,未加药组为对照。培养一定时间后 ,测定培养上清液的钙含量 ,将实验组钙含量与对照组钙含量比值作骨吸收指数。结果 :ASON( 5.0m/L)分别作用 12、2 4、48h后 ,颅盖骨骨吸收指数呈时间依赖性降低。作用 48h后 ,ASON使颅盖骨骨吸收指数呈剂量依赖性降低 ;IL -6则使其呈剂量依赖性升高 ;ASON可轻度降低IL -6升高的骨吸收指数 ;ASON( 5μm /L)对有或无IL -6培养的颅盖骨骨吸收抑制率分别为 10 .8%、19.3 % ;NSO对骨吸收指数无影响。结论 :IL -6反义寡核苷酸对体外培养的颅盖骨骨吸收有一定的抑制作用     

姜黄素对非酒精性脂肪肝细胞胆固醇代谢的影响

[目的]探讨姜黄素对体外诱导的非酒精性脂肪肝模型细胞胆固醇代谢的影响及其机制。[方法]采用油酸诱导法建立非酒精性脂肪肝细胞模型,并用姜黄素进行处理,油红O染色观察细胞内脂质累积情况,酶法检测细胞内甘油三酯、游离胆固醇和总胆固醇含量,定量PCR检测SR-BΙmRNA和HMGCR mRNA的表达。[结果]30μg·mL^-1油酸可成功建立非酒精性脂肪肝细胞模型,姜黄素干预后,脂肪肝细胞中甘油三酯和游离胆固醇减少,细胞内脂质积累减少,而总胆固醇无显著变化。姜黄素还能降低HMGCR mRNA表达,而增加SR-BⅠmRNA表达。[结论]姜黄素能够降低脂肪肝细胞游离胆固醇,其机制与其抑制内源性胆固醇的生物合成,促进外源胆固醇进入肝细胞进行代谢有关。     

血管内皮生长因子反义寡核苷酸体外抑制皮肤血管瘤内皮细胞的增殖

目的 观察血管内皮生长因子（VEGF）反义寡核苷酸对皮肤血管瘤内皮细胞体外增殖及生物学特性的影响,并探讨其机制。方法 体外培养皮肤血管瘤内皮细胞;脂质体介导VEGF反义寡核苷酸进行转染;MTT法观察其对内皮细胞生长的抑制;免疫细胞化学技术检测VEGF的蛋白表达;流式细胞仪分析细胞周期变化。结果 VEGF反义寡核苷酸作用后的内皮细胞生长明显受到抑制,作用后48hVEGF蛋白表达明显下降,细胞周期中S期细胞数明显减少,并于72h后抑制效应逐渐减弱。结论 VEGF反义寡核苷酸能显著抑制皮肤血管瘤内皮细胞的生长,其机制是抑制细胞的增殖,而不是促进其凋亡。     

增殖细胞核抗原反义寡核苷酸对裸鼠肝癌生长的影响

目的 探讨增殖细胞核抗原（PCNA）反义寡核苷酸对裸鼠人肝癌生长的抑制作用。方法 接种传代培养的人肝癌细胞于裸鼠皮下,接种后24h内局部注射PCNA反义寡核苷酸进行治疗。观察裸鼠的体重变化、瘤体大小,计算抑瘤率。HE染色观察肝癌组织结构和细胞形态改变。结果 PCNA反义寡核苷酸治疗组裸鼠肝癌的生长受到抑制,抑瘤率为72．8％。瘤重、肿瘤体积明显小于正义寡核苷酸治疗组及对照组。镜下见PCNA反义寡核苷酸治疗组肝癌细胞部分细胞形态呈梭形,核分裂相较少见,细胞的异型性较小。结论 PCNA反义寡核苷酸能显著抑制裸鼠肝癌的生长。     

阳离子脂质体介导端粒酶反义寡核苷酸促进对HeLa细胞生长的抑制作用

目的 考察阳离子脂质体(CL)为载体的反义寡核苷酸(ASODN)序列对HeLa细胞的抑制作用.方法 选择以人端粒酶模板RNA(hTR)和人逆转录酶催化亚基(hTERT)为靶点的两种ASODN,用3β[N-(N‘,N‘-二甲氨基乙基)氨甲酰基]-胆固醇(DC-Chol)和二油酰基磷脂酰乙醇胺(DOPE)制备CL,考察其形态和粒径分布,并用其介导反义寡核苷酸转染HeLa细胞,通过噻唑蓝比色法(MTT)研究对细胞的抑制作用.结果 CL外观呈圆形,粒径均匀,在体外能够有效转染反义hTR和反义hTERT寡核苷酸序列(anti-hTR,anti-hTERT).CL/ASODN处理后的HeLa细胞,72 h后的存活率为(19.82±12.19)%(antihTR,2.0 μmol·L-1)和(16.59±4.10)%(anti-hTERT,2.0 μmol·L-1),而相同浓度游离的ASODN没有发现抑制作用.CL/ASODN复合物能够在120 h内持续抑制HeLa细胞的生长.结论 CL能够有效提高ASODN对HeLa细胞的抑制作用,作为端粒酶抑制剂的新型非病毒载体,具有良好的应用前景.     

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.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.     

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.