葡萄糖转运蛋白1基因多态性对成纤维细胞糖代谢的影响

高血糖和遗传体质因素是糖尿病肾病 (ＤＮ)的主要发病因素。研究表明 ,葡萄糖转运蛋白 1基因 (ｇｌｕｃｏｓｅｔｒａｎｓｐｏｒｔｅｒ1,ＧＬＵＴ1)ＸｂａＩ(- )等位基因与 2型ＤＮ的发生相关[1] 。ＧＬＵＴ1是肾小球系膜细胞的主要葡萄糖转运蛋白[2 ] ,负责细胞的基础代谢 ,其表达与活性可能在ＤＮ细胞糖代谢异常中起了重要作用[3] 。为了进一步揭示ＧＬＵＴ1基因多态性与其功能之间的联系 ,我们对ＧＬＵＴ1不同基因型ＤＮ和正常人皮肤成纤维细胞葡萄糖摄入率和动力学 ,ＧＬＵＴ1的表达及细胞表型进行了研究。一、对象分别对 5 4名汉族健康…     

雷公藤内酯对L_(1210)细胞补救通路DNA合成的影响

用[~3 H]TdR参入法发现雷公藤内酯对L_(1210)细胞TdR转运有较强的抑制作用,而对TdR磷酸化的影响较弱。通过分析药物对DNA溶液粘滞性的影响、药物对DNA平衡透析及药物与DNA作用后的紫外光谱与荧光光谱变化等,均未发现药物与DNA之间存在相互作用。从药物对无细胞体系DNA合成无明显影响可进一步证实该药对DNA模板功能及DNA聚合酶α均无明显影响。因此对补救通路DNA合成的抑制,主要缘于该药对细胞核苷转运的抑制。     

抑制自噬增强门冬酰胺酶对小细胞肺癌细胞H1688和H446的抗癌作用

目的 研究门冬酰胺酶(asparaginase,AN)对小细胞肺癌H1688和H446细胞的抗癌作用,探讨自噬在AN抗癌过程中的功能.方法 采用MTT法及台盼蓝染色法检测AN单独或联用自噬抑制剂氯喹(chloroquine,CQ)对H1688和H446细胞的生长抑制作用;免疫荧光法观察自噬标记物LC3表达,示踪自噬体形成;Western blot检测自噬相关蛋白LC3及Akt/mTOR信号通路的表达.结果 AN呈浓度依赖性抑制H1688和H446细胞增殖并促进其死亡(P＜0.05);AN处理组可以显著增加H1688和H446细胞自噬小体数量并诱导LC3-Ⅱ表达;自噬抑制剂CQ提高AN对H1688和H446细胞杀伤作用(P＜0.05);AN作用H1688细胞后p-Akt、p-mTOR、p-70S6K蛋白表达降低.结论 AN对小细胞肺癌H1688和H446细胞具有抑制作用并诱导细胞保护性自噬,阻断自噬可以增强AN对小细胞肺癌H1688和H446细胞的抗癌疗效.     

普罗布考对动脉粥样硬化兔HDL功能的影响及其调控机制

目的观察普罗布考对动脉粥样硬化兔巨噬细胞、肝细胞HDL功能的影响,并进一步探讨普罗布考抗动脉粥样硬化的机制。方法18只新西兰大白兔随机分为:(1)正常对照组(n=6):给予普通饮食喂养;(2)动脉粥样硬化组(n=6):饲以高脂饲料;(3)普罗布考组(n=6):在饲以高脂饮食基础上给予普罗布考[400mg/(kg·d)]。12周后取血以酶法测定血脂;利用流式细胞仪检测腹腔巨噬细胞、肝细胞细胞表面ABCA1、SR-BⅠ表达量;液闪计数仪检测其[3H]胆固醇转出率。结果实验12周后,普罗布考组血清总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)、非高密度脂蛋白(NHDL-C)水平明显下降。普罗布考上调动脉粥样硬化兔腹腔巨噬细胞及肝细胞ABCA1及SR-BⅠ蛋白质表达水平。普罗布考组腹腔巨噬细胞、肝细胞[3H]胆固醇转出率较动脉粥样硬化组明显上升。结论虽然普罗布考降低HDL-C,但是其通过上调腹腔巨噬细胞、肝细胞SR-BⅠ及ABCA1的表达及增加其[3H]胆固醇转出率,改善HDL功能,具有促进动脉粥样硬化兔胆固醇逆转运(RCT)作用。     

应用RNA干扰技术检测ASCT2功能

目的谷氨酰胺(glutamine,GLN)是肠道细胞的主要氧化燃料,必须通过载体的转运才能进入细胞内,其中载体ASCT2起重要作用,文中将应用RNA干扰(RNA interference,RNAi)技术研究肠黏膜上皮细胞株(intestinal epithelial cell,IEC)-6细胞膜GLN载体ASCT2的转运功能。方法体外培养IEC-6,RNAi慢病毒颗粒进行细胞感染,3 d后荧光显微镜下观察绿色荧光蛋白(green fluorescent protein,GFP)表达情况,5 d后采用RT-PCR检测载体ASCT2的mRNA表达,7 d后采用Western blot检测其蛋白表达,同时测定[3 H]-L-GLN摄取率。结果病毒转染细胞中均有GFP表达,载体ASCT2的mRNA表达下降60%,蛋白表达几近消失,[3H]-L-GLN摄取率下降15%。结论通过慢病毒载体介导的RNAi这一方便有效的方法,证实了ASCT2载体在整个细胞的GLN转运中起15%以上的作用。     

血小板激活因子对小鼠脾细胞增殖以及产生IL—2,NKCF和CSF的影响

本文研究了PAF对BALB/c小鼠脾细胞增殖以及产生IL-2,NKCF和CSF的影响。[~3H]TdR掺入法检测脾细胞增殖结果表明PAF在10~(-10)～10~(-7)mol/L范围内显著抑制ConA诱导的脾细胞增殖。用CTLL-2细胞测定IL-2,YAC-1细胞检测NKCF,小鼠骨髓细胞测定CSF,结果表明当PAF浓度为10~(-7)mol/L时,三种因子的产生均被明显抑制。PAF对小鼠脾细胞功能的抑制可能与其广泛的病理生理作用有关。     

细梗香草总皂苷诱导肺癌细胞H460凋亡及其机制的研究

目的：观察细梗香草总皂苷（LC）对非小细胞肺癌H460细胞增殖和凋亡的影响。方法在H460肺癌细胞培养时加入不同浓度的LC,采用MTT法和克隆形成实验检测细胞的增殖活性,AnnexinⅤ/PI（FCM）法观察LC对肺癌细胞凋亡的诱导作用和周期分布的影响。DCFH- DA荧光探针FCM检测细胞内活性氧生成,Western blot法检测NF-κB、I-κB、Bax、Bcl-2、Capase-3的蛋白表达。结果 LC可抑制H460细胞生长,具有剂量、时间依赖性;LC作用H460细胞后克隆形成率明显下降。LC可引起H460细胞凋亡,并可将H460生长阻滞在S期。LC以浓度依赖方式增高H460细胞内活性氧水平,还可使H460细胞I-κB、Bax和Capase-3表达增加,而使NF-κB和Bcl-2表达降低。结论 LC可以抑制H460细胞增殖,可能是通过增加细胞内活性氧表达,激活了肺癌细胞线粒体凋亡途径。     

用快速比色计量法测量细胞生长

有许多方法可用来检测体外培养细胞的生长、增殖或抑制(如染料排除法、软琼脂克隆试验和[~3H]胸腺嘧啶渗入法等),用于对增殖或活性因子和抗肿瘤药物筛选的研究,但这些方法因重复性差、实验周期长或操作困难等缺点,不宜推广。Mosmann报道的比色试验,检测培养于96孔板的活细胞数,其原理是活细胞脱氨酶的催化作用能使四唑盐(MTT)还原成蓝色甲臢结晶,后者溶于有机溶剂,进而用ELISA仪检测吸收光密度     

桂枝茯苓丸逆转lncRNA H19介导的miR-195-5p/SGK1通路治疗子宫内膜癌机制研究

目的探究桂枝茯苓丸对子宫内膜癌Ishikawa细胞的作用和调控机制。方法进行实时定量PCR(qRT-PCR)检测lncRNA H19在人正常子宫内膜细胞EMC和子宫内膜癌细胞HEC-1A、HEC-1B、Ishikawa细胞表达。向Ishikawa细胞转染sh-H19,利用细胞计数试剂盒8(CCK8)检测细胞生长活力,流式细胞术评估细胞凋亡率,qRT-PCR和蛋白免疫印迹法(Western blot)分析自噬标志蛋白P62、Beclin1和LC3II的表达。采用双荧光素酶报告基因检测验证lncRNA H19与miR-195-5p及miR-195-5p与SGK1的靶向结合关系。将oe-H19转染到Ishikawa细胞,并将转染oeH19的Ishikawa细胞在桂枝茯苓丸含药培养液中培养。qRT-PCR检测lncRNA H19表达,CCK8、流式细胞术检测细胞的增殖和凋亡,Western blot检测自噬相关蛋白P62、Beclin1和LC3II的表达。结果与人正常子宫内膜细胞EMC (1.00±0.09)比较,lncRNA H19在子宫内膜癌细胞HEC-1A(1.73±0.13)、HEC-1B(2.12±0.14)、Ishikawa(3.18±0.16)表达显著升高(P0.05或P0.01)。与对照组比较,下调lncRNA H19使子宫内膜癌细胞增殖活性显著降低[(45.00±3.95)%比(100.00±8.12)%,P0.05],细胞凋亡率明显升高[(10.06±1.12)%比(5.35±0.55)%,P0.05]。敲减lncRNA H19的子宫内膜癌细胞P62表达显著降低,Beclin1和LC3II表达显著上调。双荧光素酶报告基因检测和Western blot结果表明,lncRNA H19靶向负调控miR-195-5p表达,miR-195-5p靶向抑制SGK1表达。在挽救实验中,与oe-NC组(1.00±0.09)比较,oe-H19组(3.23±0.29)Ishikawa细胞lncRNA H19含量显著增高(P0.01),而桂枝茯苓丸能够逆转oe-H19转染引起的lncRNA H19水平上升(1.14±0.12)(P0.05)。oe-H19组[(240.00±15.89)%]细胞存活率较oe-NC组[(100.00±7.88)%]显著增高(P0.05),转染oe-H19后的Ishikawa细胞经桂枝茯苓丸含药培养液处理后,其存活率明显降低(92.00±8.47)%(P0.05)。oe-H19转染提高Ishikawa细胞P62表达,抑制Beclin1和LC3II表达,经桂枝茯苓丸含药培养液处理,Ishikawa细胞P62 Beclin1和LC3II表达部分恢复。结论桂枝茯苓丸具有抗子宫内膜癌活性,其作用机制可能是抑制lncRNA H19介导的miR-195-5p/SGK1通路,进而抑制子宫内膜癌细胞的增殖,促进细胞自噬。     

地冬鳖甲煎剂对小鼠甲亢模型肝组织~3H—亮氨酸及~3H—胸腺嘧啶核苷摄入的影响

本实验采用常用的养阴药物生地、麦冬、鳖甲治疗小鼠甲亢模型。观察其对肝组织~3H—亮氨酸和~3H—胸腺嘧啶核苷摄入的影响。实验发现,甲亢组肝组织~3H—亮氨酸摄入率低于正常组,差别有高度显著性(P<0.01)。甲亢加中药治疗组摄入率提高,与正常组比较差别无显著性(P>0.05)。另方面,甲亢组~3H—胸腺嘧啶摄入率高于正常组,差别有高度显著性(P<0.01)。甲亢加中药治疗组摄入率下降至正常,与正常组比较差别无显著性(P>0.05)。结果提示,养阴药物生地、麦冬、鳖甲有调节蛋白质和DNA代谢的作用。     

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.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

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.