高糖、高胰岛素及二甲双胍对人乳腺癌MCF-7细胞增殖的影响

目的：探讨高糖、高胰岛素和不同浓度二甲双胍对乳腺癌MCF-7细胞增殖的影响。方法：体外培养人乳腺癌细胞株MCF-7,分别给予高糖和（或）高胰岛素及不同浓度的二甲双胍进行干预,MTT法检测MCF-7细胞的增殖情况。结果：高糖、高胰岛素、高糖高胰岛素作用不同时间均可促进MCF-7细胞增殖;不同浓度的二甲双胍干预后,MCF-7细胞的生长均受到一定程度的抑制,且随时间和浓度的增加而增加。结论：不同浓度二甲双胍可对高糖、高胰岛素作用下乳腺癌MCF-7细胞的增殖起到抑制作用,且呈时间和浓度依赖性。     

胰岛素及其类似物单独或联合二甲双胍对人乳腺癌细胞系 (MCF-7)增殖的影响

目的:探讨人胰岛素及其类似物对人乳腺癌细胞系(MCF-7)增殖的影响及二甲双胍联合胰岛素及其类似物对MCF-7细胞增殖的影响?方法:①使用不同浓度(0?1?10?100?1 000 nmol/L)的人胰岛素及类似物干预MCF-7细胞72 h 后,用CCK-8法检测干预后细胞增殖情况?②用胰岛素及类似物(每种胰岛素浓度均为1 000 nmol/L)干预MCF-7细胞,分别用CCK-8法检测各胰岛素在干预24?48?72 h后的细胞增殖?③用CCK-8法检测单纯二甲双胍(0?2.5?5.0?10.0?20.0 mmol/L),及二甲双胍联合胰岛素及其类似物(浓度均为1 000 nmol/L)干预 MCF-7 72 h后细胞的增殖情况?结果:人胰岛素及类似物均能促进乳腺癌细胞系增殖并呈时间依赖性,人胰岛素?甘精胰岛素?赖脯胰岛素对细胞的增殖作用呈显著的剂量依赖性,相同条件下不同胰岛素对细胞的增殖作用无明显差异,但甘精胰岛素?赖脯胰岛素在相同条件下促细胞增殖作用高于其他胰岛素?二甲双胍可以呈浓度依赖性地抑制细胞的增殖作用,并减弱胰岛素类似物引起的细胞增殖作用,相同浓度二甲双胍对不同胰岛素类似物促增殖的抑制作用无明显差异,但对地特胰岛素的抑制作用小于其他胰岛素类似物?结论:胰岛素类似物和人胰岛素均能促进肿瘤细胞的增殖,二甲双胍可以抑制MCF-7细胞增殖,二甲双胍可以拮抗胰岛素及其类似物引起的促细胞增殖作用?     

二甲双胍对人乳腺癌ZR75—1细胞增殖及凋亡的调控

目的探讨抗糖尿病药物二甲双胍对人乳腺癌ZR75—1细胞增殖抑制及凋亡诱导的作用。方法3种不同浓度的二甲双胍处理乳腺癌ZR75—1细胞24、48h,MTT法检测ZR75-1细胞的增殖活性,流式细胞仪（FcM）检测48h后各浓度二甲双胍诱导细胞凋亡情况及对细胞周期的影响。结果与对照组相比,二甲双胍以时间、剂量依赖性方式抑制ZR75—1细胞增殖（P〈0．05）。作用48h后,二甲双胍诱导ZR75—1细胞凋亡并阻断了细胞周期进程,凋亡率随二甲双胍浓度的增加而增加（P〈0．05）,且随着药物浓度的增加,G0／G1期细胞比例逐渐增加,S期及G2／M期细胞比例逐渐下降（P〈0．05）。结论二甲双胍可显著抑制ZR75—1细胞的增殖,诱导其凋亡,阻滞细胞周期进程。     

二甲双胍对人肝细胞癌SMMC7721增殖的抑制作用

[摘要]目的检测二甲双胍作用下肝细胞癌SMMC-7721细胞的增殖和凋亡情况,并初步探讨了其中的机制。方法以不同浓度(0.2、1、2、5 mmol/L)二甲双胍处理SMMC-7721,用CCK-8法检测其对细胞增殖的影响,Hochest33342 DNA染色检测细胞凋亡,实时定量PCR法检测Bcl-2和Bid mRNA的表达情况。结果二甲双胍处理48 h 和96 h后,人肝细胞癌SMMC7721细胞的增殖明显受到抑制(P<0.05),以2 mmol/L和5 mmol/L组最为明显(P<0.01),并呈时间和剂量依赖性。Hochest33342染色发现,1 mmol/L和5 mmol/L浓度的二甲双胍可促进细胞凋亡,细胞凋亡率与对照组相比差异有统计学意义(P<0.05)。二甲双胍处理后,SMMC7721细胞内抗凋亡分子Bcl-2表达下降(P<0.01),促凋亡分子Bid表达升高(P<0.05)。结论二甲双胍能抑制人肝细胞癌SMMC7721细胞增殖,促进细胞凋亡,其机制可能与细胞内抗细胞凋亡分子Bcl-2表达下降及促凋亡分子Bid表达升高有关。     

二甲双胍对人血清诱导的胰腺癌细胞增殖及MMPs表达的影响

目的：探讨二甲双胍对糖尿病患者血清诱导的胰腺癌细胞株Patu8988增殖的影响及相关基因表达的变化。方法：以糖尿病患者血清诱导细胞株Patu8988为糖尿病组,以健康人血清为对照组,用不同浓度的二甲双胍（2.5、5.0、10.0、20.0、40.0 mmol/L）干预两组细胞,通过CCK-8细胞增殖实验检测不同血清诱导后细胞的增殖能力,采用RT-PCR检测二甲双胍干预后基质金属蛋白酶（MMP）-2、MMP-8和MMP-11 mRNA表达的差异。结果：与健康对照组相比,糖尿病患者血清诱导后促进Patu8988细胞增殖（0.74±0.03 vs.0.62±0.02,P＜0.01）;二甲双胍抑制糖尿病患者血清诱导的细胞增殖（P＜0.01）;RT-PCR结果显示二甲双胍减弱MMP-2、MMP-8、MMP-11的mRNA表达。结论：二甲双胍抑制糖尿病患者血清诱导的细胞增殖,这种抑制效应可能与其对MMP-2、MMP-8、MMP-11mRNA表达的下调作用有关。     

二甲双胍对结肠癌细胞增殖、周期及凋亡的影响

  目的 研究二甲双胍对人结肠癌细胞株SW480增殖、周期及凋亡的影响,并初步探讨其可能的机制。方法 体外培养人结肠癌细胞株SW480,给予不同浓度二甲双胍干预,MTT法检测细胞增殖情况,流式细胞仪检测细胞周期及凋亡,Real time-PCR法检测相关基因mRNA的表达。结果 二甲双胍可以抑制SW480细胞的增殖,并呈时间-剂量依赖性。20mmol/L二甲双胍干预48h后与对照组相比,细胞G0/G1期比例升高,S及G2/M期比例均降低;细胞凋亡比率增高;Real time-PCR示周期相关基因CyclinD1 mRNA表达明显下调（P0.05),凋亡相关基因BCL-2 mRNA表达明显下调,CASP3及BAX mRNA 表达明显上调(P<0.01)。结论 二甲双胍能够抑制人结肠癌细胞株SW480的增殖,G0/G1期细胞周期阻滞,促进细胞凋亡;其机制可能与上调及下调某些周期、凋亡相关基因的mRNA表达有关。     

二甲双胍可能通过Hippo-YAP通路抑制HER-2阳性乳腺癌细胞的增殖和促进凋亡

目的 观察二甲双胍对HER-2阳性乳腺癌细胞株SKBR3增殖、凋亡的影响,并探讨其可能的机制。方法 分别用0、20、40、60、80、100、120 μmol/L二甲双胍处理乳腺癌细胞株SKBR3,CCK-8法检测其对细胞增殖的影响;结晶紫染色观察其对细胞集落形成的影响;计算IC50值。以该浓度二甲双胍处理细胞,与空白对照相比,流式细胞术检测细胞凋亡和周期的改变;实时荧光定量 PCR（qRT-PCR）检测 YAP、TAZ、EGFR、CTGF、CYR61、E-cadherin、N-cadherin、Vimentin 及 Fibronectin 等关键基因的mRNA水平表达;Western blotting 实验检测YAP、TAZ蛋白水平的表达;免疫荧光观察二甲双胍对SKBR3细胞中YAP/TAZ核易位的改变。结果 CCK-8和细胞克隆实验显示二甲双胍对SKBR3细胞增殖活力具有明显的抑制作用（P<0.05）,呈浓度和时间依赖性。与对照组相比,二甲双胍处理后的SKBR3细胞凋亡明显增加;G1期细胞比例增多,而G2/M期细胞比例减少。N-cadherin、Vimentin和Fibronectin的表达降低,而E-cadherin的表达上调（P<0.05）。qRT-PCR和Western blotting检测结果显示,二甲双胍处理后YAP、TAZ、EGFR、CTGF和CYR61的表达明显下调（P<0.05）。免疫荧光实验显示：实验组较对照组的YAP或TAZ,其荧光的定位更多地聚集于胞浆,而细胞核中荧光的量明显的减少。结论 二甲双胍能抑制HER-2阳性乳腺癌细胞SKBR3的增殖,促进其凋亡和上皮－间质转化,其机制可能与抑制YAP/TAZ的表达和核定位有关。     

二甲双胍对肝癌细胞HCCLM3生物学行为的影响

目的 研究二甲双胍对人肝癌细胞株HCCLM3增殖、凋亡及迁移能力的影响.方法 体外培养人肝癌细胞株HCCLM3,以不同浓度的二甲双胍干预细胞,CCK-8法检测细胞增殖情况,RT-PCR检测Bax及Bcl-2基因表达,细胞划痕实验观察细胞迁移情况.结果 随着二甲双胍浓度增高和作用时间延长,对HCCLM3细胞增殖抑制率较对照组显著提高(P＜0.05);浓度达到10 mmol/L可抑制HCCLM3细胞迁移能力;作用48h后HCCLM3细胞中Bax表达呈现明显递增趋势,而Bcl-2表达呈现递减趋势.结论 二甲双胍能够以浓度及时间依赖性的形式抑制肝癌细胞HCCLM3细胞的增殖及迁移活动,并能够影响其凋亡相关基因Bax与Bcl-2的表达.     

二甲双胍对类风湿性关节炎患者滑膜细胞增殖及凋亡的影响

目的 探讨二甲双胍对类风湿性关节炎患者滑膜细胞增殖与凋亡的影响.方法 取类风湿性关节炎患者滑膜组织,用胰酶消化法分离其滑膜细胞,分别用流式细胞术和CCK-8法检测三种浓度的二甲双胍(10、20、30 mmol/L)对滑膜细胞增殖的影响,TUNEL和流式细胞仪检测滑膜细胞凋亡情况,ELISA法检测二甲双胍处理的滑膜细胞上清中的炎性因子和抗炎性因子.结果 三种浓度(10、20、30 mmol/L)的二甲双胍分别干预滑膜细胞24、48、72 h,均不同程度抑制了滑膜细胞的增殖,且成时间-浓度依赖性.CCK-8法检测结果显示20、30 mmol/L二甲双胍处理显著抑制了滑膜细胞的增殖(P＜0.05),流式细胞术结果表明20 mmol/L二甲双胍处理48 h后,Go/G1期滑膜细胞的百分比与对照组相比显著增加了约20％ (P ＜0.05),S期和G2/M期的细胞比例分别显著下降了9％和11％ (P ＜0.05).凋亡结果表明,20、30 mmol/L二甲双胍处理48、72 h,较对照组显著促进滑膜细胞的凋亡(P＜0.05),流式细胞术显示早期和晚期的滑膜细胞凋亡率约17.5％.检测炎性因子发现,20、30 mmol/L二甲双胍处理后显著降低炎症因子IL-6、IL-1β和TNF-α的水平(P＜0.05),增加抗炎因子IL-10水平约66％和76％ (P ＜0.05).结论 二甲双胍可显著抑制体外培养的人滑膜细胞的增殖,诱导其凋亡,可能与二甲双胍降低炎性因子TNF-α、IL-6、IL-1β水平和增加抗炎因子IL-10水平有关.     

二甲双胍抑制人前列腺癌细胞上皮间质转化及其作用机制

目的 研究二甲双胍对前列腺癌Vcap细胞增殖、侵袭及上皮间质转化（EMT）的影响,初步探讨microRNA(miRNA) 相关的作用机制。方法 以PBS处理组作为对照,使用不同浓度二甲双胍(1~50mmol/L) 处理前列腺癌Vcap细胞,MTS比色法检测细胞的增殖能力;流式细胞术分析二甲双胍对Vcap细胞周期分布的影响;划痕和侵袭小室实验分别检测5mmol/L二甲双胍和miR30a对细胞迁移和侵袭能力的作用;使用RT-PCR和Western blotting测定5mmol/L二甲双胍对Vcap细胞上皮指标物（E-cadherin、β-catenin）和间质指标物（Vimentin、Snail）mRNA和蛋白表达水平的影响;使用RT-PCR检测miR30a、miR143、miR185、miR196、miR205的表达水平变化。结果 二甲双胍抑制前列腺癌Vcap细胞的增殖,且呈浓度和时间依赖性。5mmol/L 二甲双胍明显影响Vcap细胞的周期分布并显著抑制Vcap细胞的迁移和侵袭能力。二甲双胍在mRNA和蛋白水平上,显著上调Vcap细胞中E-cadherin 和β-catenin的表达(P均<0.05) ,下调Vimentin和N-cadherin的表达(P均<0.05)。进一步的实验发现,二甲双胍显著上调miR30a的表达水平 (P<0.05),而后者可显著抑制Vcap细胞的增殖和EMT的发生。结论 二甲双胍明显抑制前列腺癌Vcap细胞的增殖、侵袭能力和上皮间质转化的过程。该过程可能涉及二甲双胍对miR30a的表达的上调。     

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.     

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.     

Electro-Acupuncture Combined with the Trigger Point Needle-Embedding for Treatment of Primary Trigeminal Neuralgia in 31 Cases

In recent years, the author of this essay has applied electro-acupuncture combined with the trigger point needle-embedding for treatment of primary trigeminal neuralgia in 31 cases, yielding satis- factory results as reported in the following.     

Investigation of Matrix Metalloproteinase -1, 2 and tissue inhibitor of matrix metalloproteinases-1 in Endometriosis

Objective： To explore the role of matrix metalloproteinase-1,2 （MMP-1, MMP-2） and tissue inhibitor of matrix metalloproteinases-1 （TIMP-1） in endometriosis. Methods： The eutopic and ectopic endometria from 40 subjects suffering from endometriosis and regular.endometria from 40 subjects （excluding endometriosis） were collected and examined by in situ hybridization technology and western blot assay. Results： Both expressions of MMP-1 and -2 were stronger in ectopic endometrium and eutopic endometrium than in normal endometrium. On the contrary, the expression of TIMP-1 in ectopic endometrium and eutopic endometrium was lower. The differences were significant （P 〈 0.01 ）. Moreover, there was no relationship among the expressions of MMP-1, 2 and TIMP-1 in ectopic endometrium. Conclusion： The expressions of MMP-1, 2 and TIMP-1 lose balance and lack of periodic changes in ectopic endometrium , which explains the biological invasive behavior of endometriosis. It was suggested-that regulating the balance between the MMPs and TIMP-1 should be an ideal therapeutic target to endometriosis.     

Shi Da-zhuo (史大卓) —An outstanding cardiologist of integrative medicine

Prof. SHI Da-zhuo, Ph.D., male, was born on March 20, 1960. Prof. SHI entered the Ph.D. program in 1990 at the China Academy of Chinese Medical Sciences under the supervision of Prof. CHEN Ke-ji, majoring in the treatment of cardiovascular diseases. After receiving his Ph.D. degree in 1993, Prof. SHI started working at the Cardiovascular Center in Xiyuan Hospital affiliated to China Academy of Chinese Medical sciences.     

Persist in and Carry Forward the Integrative Medical Research——Speech at the 6th National General Congress of Chinese Association of Integrative Medicine by Academician CHEN Zhu,Minister of Ministry of Health

The 6th National General Congress of Chinese Association of Integrative Medicine （CALM） was convened at 19-20, April 2008 in Beijing. Academician CHEN Zhu, the minister of Ministry of Health indicated at the congress that the integration of Chinese and Western medicine is very well in keeping with the situation of our country and the general rule of development in medical science; and as a good integration of Chinese medicine and Western medicine, it is mutually beneficial and advantageous to both of them. Seeing the creativity shown in integrative medical investigation in theoretic and methodological sides, we should and must persist in and develop it.