辛伐他汀对酒精诱导大鼠骨髓基质细胞成脂及成骨基因表达的影响

目的观察辛伐他汀对酒精诱导大鼠骨髓基质细胞成脂及成骨基因改变的影响,探讨辛伐他汀治疗酒精性骨坏死的可能性。方法分离培养SD大白鼠骨髓基质细胞（MSCs）,随机分为三组：模型组：第1周以含终浓度0．09mol／L酒精的培养液培养,第2周停用酒精,实验组：第1周以含终浓度0．09mol／L酒精的培养液培养,第2周停用酒精,改以含终浓度1×10^-7mol／L辛伐他汀的培养液培养;对照组：全程以不加酒精和辛伐他汀的培养液培养。于实验第14天时收集细胞,以RT—PCR检测细胞中过氧化物酶体增殖子激活受体-γ（PPARγ）mRNA和骨钙素（OC）mRNA的表达。实验数据以均数±标准差（x^-±S）表示,采用SPSS10．0软件进行统计学分析,多组间比较采用单因素方差分析,样本均数间的两两比较采用LSD—t检验,检验水准仪=0．05。结果与对照组比较,模型组细胞PPARγmRNA表达水平明显升高（P〈0．05）,实验组细胞PPARγmRNA表达水平稍高（P〉0．05）;与对照组比较,模型组OCmRNA表达水平明显降低（P〈0．05）,实验组OCmRNA表达水平稍低（P〉0．05）。结论辛伐他汀能抑制酒精诱导MSCs成脂基因表达,对抗或解除酒精对MSCs的成骨分化的抑制而增加成骨基因表达,可能有治疗酒精性骨坏死的作用。     

辛伐他汀对骨髓基质干细胞成骨分化晚期骨钙素表达的影响

目的通过辛伐他汀体外干预成骨分化晚期的骨髓基质干细胞,观察骨钙素（osteocalcin,OCN）在mRNA和蛋白水平的表达,探讨辛伐他汀促进成骨分化作用及机制。方法取4周龄大鼠股骨和胫骨骨髓腔内的干细胞体外培养,传一代后并向成骨细胞诱导分化,取第五代细胞分别加入lOl7M浓度的辛伐他汀或等量溶剂干预,于加药后48h提取细胞RNA及蛋白,采用Realtime PCR和Westernblot法检测0CN的表达,同时收集细胞培养液上清,ELISA法检测0CN浓度;1周后采用yonKossa染色观察细胞外基质矿化能力。结果①RealtimePCR：实验组0CN的mRNA表达水平显著高于对照组（P〈0．05）;②Westernblot：实验组OcN的蛋白表达水平显著高于对照组（P〈0．05）;③ELISA：实验组细胞培养上清中0CN浓度显著高于对照组（P〈0．05）;④VonKossa染色：实验组细胞外基质矿化能力强于对照组。结论辛伐他汀可以刺激大鼠骨髓基质干细胞向成骨细胞分化晚期0CN在rnRNA和蛋白水平的表达。     

酒精性骨坏死发病机制的分子生物学实验研究

目的 :观察酒精诱导骨髓基质细胞 (Marrowstromalcells ,MSCs)的作用 ,揭示酒精性股骨头坏死新的发病机制。方法 :采用原代MSCs体外培养技术 ,取小鼠股骨骨髓细胞 ,分离获取MSCs。以酒精作为脂肪细胞分化诱导剂处理细胞 ,以苏丹Ⅲ染色 ,光镜下计数脂肪细胞。并测定细胞内甘油三酯、碱性磷酸酶活性和培养液中骨钙素含量。采用完整细胞斑点印迹分子杂交方法检测酒精 ( 0 .0 9mol/L)组和对照组细胞中 42 2(aP2 )mRNA和Ⅰ型胶原mRNA的表达。结果 :以递增浓度酒精 ( 0 .0 3、0 .0 9、0 .15mol/L)处理细胞 2 1d ,MSCs分化生成的脂肪细胞数量随酒精作用时间延长及浓度增大而增多 ,具有一定时间、剂量依赖性。与对照组相比 ,细胞内甘油三酯含量明显增高 ,碱性磷酸酶活性随酒精浓度增大而降低 ,培养液中骨钙素含量显著减少。实验组中 42 2 (aP2 )mRNA表达含量 72 0 7.8± 3 3 1.3 ,显著高于对照组 65 2 .2± 62 .6,P <0 .0 0 1。其Ⅰ型胶原mRNA表达含量 3 5 67.3± 3 0 0 .9,明显低于对照组 7487.0± 488.4,P <0 .0 0 1。结论 :酒精能够从基因调控水平诱导MSCs大量分化为脂肪细胞 ,成骨分化减少 ,这可能是长期酗酒后股骨头骨髓内脂肪组织增多 ,骨内压增加 ,血流灌注减少 ,导致缺血 ,同时骨修复能力不足 ,从而发生     

地塞米松调节骨髓基质细胞成脂及成骨分化的组织化学研究

目的：观察地塞米松对骨髓基质细胞分化成脂肪细胞的影响,探讨激素性股骨头缺血性坏死的发病机制。 方法：骨髓基质细胞在含有不同浓度地塞米松的培养基中培养,苏丹Ⅲ染色作脂肪细胞计数。采用生化分析仪 测定细胞内甘油三酯含量及碱性磷酸酶活性。结果：脂肪细胞数随地塞米松浓度增大及作用时间延长而增多。 实验组甘油三酯含量明显高于对照组。碱性磷酸酶活性随地塞米松浓度增大而降低。结论：地塞米松能够直接 诱导骨髓基质细胞分化成脂肪细胞,减少其向成骨细胞分化,这可能与激素性骨坏死的发病机制有关。     

条件培养液对兔骨髓基质细胞向成骨细胞分化的影响

目的 :观察条件培养液对兔骨髓基质细胞向成骨细胞分化的影响。　方法 :分离兔骨髓 ,培养骨髓基质细胞 ,传代细胞分别用标准培养液和加入 β 甘油磷酸钠 ,地塞米松和抗坏血酸制成的条件培养液培养 ,观察不同培养液下骨髓基质细胞碱性磷酸酶活性和骨钙素含量。　结果 :在条件培养液下 ,骨髓基质细胞碱性磷酸酶活性和骨钙素含量明显高于标准培养液下的含量。　结论 :条件培养液有助于骨髓基质细胞向成骨细胞的分化和增殖     

酒精对骨髓多能干细胞PPARγ及骨钙素mRNA表达的影响

目的 :观察酒精对骨髓多能干细胞 (D1细胞 )成脂转录因子 (PPARγ)mRNA和成骨基因骨钙素mRNA表达的影响。方法 :以不同浓度 (0 (对照组 ) ,0 .0 9,0 .15 ,0 .2 1mol/L)酒精处理D1细胞 14d ,苏丹ⅠV染色 ,采用计算机图像分析软件确定脂肪细胞百分比 ,采用RT PCR技术检测细胞中PPARγ及骨钙素mRNA表达。结果 :对照组脂肪细胞生成很少 ,酒精处理组脂肪细胞生成显著增多 ,脂肪细胞百分比随酒精浓度增大而增多 (P <0 .0 0 1)。各组与对照组PPARγmRNA表达差异无统计学意义。 0 .0 9,0 .15 ,0 .2 1mol/L酒精组骨钙素mRNA表达降低 ,分别比对照组减少 38%、4 3%和 5 4 % (P <0 .0 0 1)。结论 :酒精能够导致骨髓多能干细胞大量分化为脂肪细胞 ,可能是酒精在脂肪酸旁路代谢下游所起的作用 ,这可能与酒精性骨坏死的发生机制有关。     

乙醇对人骨髓间充质干细胞成脂转录因子γ和骨钙素mRNA表达的影响

目的:观察乙醇对人骨髓间充质干细胞(hBMSCs)内成脂转录因子(PPAR)γ和骨钙素(Osteocalcin)mRNA表达的影响.方法:取健康自愿者骨髓,通过梯度离心、贴壁分离培养,获得hBMSCs,传至第2代hBMSCs 8 d,随机分为2组,实验组以0.09 mol/L乙醇作为诱导剂每d处理细胞,对照组不给予乙醇,继续培养细胞6 d.应用RT-PCR技术检测2组细胞中PPARγ mRNA和Osteocalcin mRNA的表达.结果:与对照组比较,实验组hBMSCs内PPARγ mRNA表达增高(1.38±0.27 vs 1.73±0.34,P＜0.05).与对照组比较,实验组hBMSCs内Osteocalcin mRNA表达降低(1.08±0.22 vs 0.48±0.11,P＜0.05).结论:乙醇能够诱导hBMSCs PPARγ表达增强,Osteocalcin表达减少,这可能与酒精性骨坏死的发生机制有关.     

芦荟大黄素对非酒精性脂肪性肝炎细胞模型脂质过氧化损伤的影响

目的：探讨芦荟大黄素（aloe-elodin,AE）对非酒精性脂肪肝炎（non-alcoholic steatohepatitis,NASH）细胞模型脂质过氧化损伤的影响。方法：将正常人肝细胞株 L-02分为对照组和实验组,对照组用普通培养基,实验组按培养基含30％脂肪乳剂终浓度不同再分为0．25,0．5,0．75,1 mL／L 组,培养72 h 后予以油红 O 染色,观察细胞形态和细胞内脂肪滴数量,测定培养液谷丙转氨酶（ALT）、谷草转氨酶（AST）、三酰甘油含量。选择最合适浓度（0．5 mL／L 组）建立 NASH 模型,再设立对照组、模型组、实验组,实验组加入10－4,10－5,10－6 mmol／L 的 AE,分别于培养24,48,72 h 监测细胞内脂滴数量、三酰甘油含量和细胞培养液中 ALT、AST、超氧化物歧化酶（SOD）、丙二醛、内毒素、肿瘤坏死因子（tumor necrosis factor alpha,TNF-α）等指标。结果：用含30％脂肪乳剂0．5 mL／L 的培养液培养 L-02细胞株72 h,成功建立 NASH 肝细胞模型。不同浓度 AE 干预 NASH 细胞72 h 后,培养液中 ALT、AST、丙二醛、内毒素、TNF-α等含量均较模型组明显下降,SOD 含量较模型组明显升高（P ＜0．05）。结论：AE 可改善肝细胞脂质过氧化程度,减轻炎症反应及细胞损伤。     

儿童骨髓基质细胞体外培养

目的 探讨儿童骨髓基质细胞体外培养条件,为儿童骨组织工程研究选择种子细胞。方法 将儿童髂骨内骨髓分别采用密度梯度离心法和全骨髓法培养,待细胞长满培养瓶后传代,选取正常生长骨髓基质细胞第3代绘制生长曲线并测定细胞内骨钙素(OCN)及培养液中碱性磷酸酶(ALP)含量。培养期间用倒置相差显微镜观察细胞形态。结果 原代儿童骨髓基质细胞呈长梭形、三角形或多角形,存在有集落样生长特性,传代后细胞形态无明显变化,且生长、增殖稳定,细胞内OCN及培养液中ALP含量在1周左右达到高峰。结论 采用密度梯度离心法和全骨髓法获得儿童骨髓基质细胞,经培养、传代后,可以得到大量稳定增殖的骨组织工程种子细胞。     

骨形态发生蛋白对人骨髓基质细胞的体外成骨分化影响

目的 提取牛骨形态发生蛋白，并观察其对人骨髓基质细胞的体外成骨分化作用，为将骨形态发生蛋白与骨髓基质细胞用于进一步的缺损修复研究提供实验依据。方法 从小牛骨中的提取骨形态发生蛋白，配制条件培养液，并作用于培养状态的人骨髓基质细胞，染色检测细胞碱性磷酸酶，I型胶原表达情况，放免法测定培养液中骨钙素含量。结果 人骨髓基质细胞经衣导培养后，碱性磷酸酶，I型胶原，骨钙素表达明显增加。结论 体外培养时，天然骨形态发生蛋白可促进人骨髓基质细胞的成骨分化。     

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.