circROBO1上调KLF5促进视网膜细胞瘤侵袭的机制研究

目的：探索circROBO1通过上调KLF5促进视网膜Y79细胞侵袭的作用及其可能的调控机制。方法：RNase R酶消化及qRT-PCR实验检测环状RNA circROBO1在视网膜Y79细胞中的结构稳定性；分别提取视网膜Y79细胞的胞质及胞核RNA，进行circROBO1的亚细胞定位分析；采用siRNA沉默视网膜Y79细胞中circROBO1的表达，划痕实验、Transwell细胞侵袭与迁移实验检测circROBO1对Y79细胞迁移与侵袭能力的影响；通过CircInteractome及TargetScan在线软件分别预测circROBO1与下游miRNA及miRNA与下游靶基因KLF5的靶向结合位点，并采用双荧光素酶报告基因实验验证两者之间的靶向调控关系。蛋白免疫印迹实验检测siRNA沉默Y79细胞中circROBO1的表达后对KLF5表达的影响。结果：与未经RNase R酶处理的对照组相比，circROBO1的相对表达量在RNase R酶处理后并无显著变化；而线性ROBO1的相对表达量在经RNase R酶处理后，则显著降低（t=16.18,P<0.05）;circROBO1在...     

circCDYL靶向miR-552-5p抑制胃癌AGS细胞的增殖和转移

目的探讨环状RNA Y染色域样(circCDYL)对微小RNA-552-5p(miR-552-5p)及胃癌AGS细胞增殖和转移的影响。方法实时荧光定量PCR分析胃癌组织、癌旁正常组织样本中circCDYL和miR-552-5p表达水平。运用CCK-8法、集落形成、划痕愈合实验和Transwell实验分析circCDYL和miR-552-5p表达对AGS细胞增殖、迁移和侵袭的影响。Western blotting分析E-cadherin、N-cadherin蛋白表达。双荧光素酶报告实验分析circCDYL和miR-552-5p靶向关系。结果与癌旁正常组织比较,胃癌组织中circCDYL表达显著降低(P＜0.05),而miR-552-5p表达显著升高(P＜0.05)。过表达circCDYL或干扰miR-552-5p后AGS细胞增殖、集落形成数、划痕愈合率、侵袭细胞数均显著降低(P＜0.05)。circCDYL直接与miR-552-5p结合,上调miR-552-5p能够逆转过表达circCDYL对AGS细胞增殖、迁移和侵袭的抑制作用(P＜0.05)。结论circCDYL通过靶向miR-552-5p抑制胃癌AGS细胞增殖、迁移和侵袭。     

miR-34a抑制人鼻咽癌细胞生长与侵袭

目的探讨miR-34a在人鼻咽癌CNE2细胞中的生物学功能。方法采用MTT、Transwell侵袭实验、细胞划痕实验分别检测过表达miR-34a对人鼻咽癌CNE2细胞的生长与侵袭作用。结果 MTT检测发现,过表达miR-34a可抑制人鼻咽癌CNE2细胞的生长增殖;细胞划痕和Transwell侵袭实验显示,在鼻咽癌CNE2细胞中过表达miR-34a 48 h后划痕愈合率为40.3%±9.0%,与对照组（76.3%±6.8%）比较,能明显减缓CNE2细胞划痕的愈合,转染miR-34a mimics 48 h后穿过基底膜的细胞数为71±10,与对照组（147±5）比较,能明显减缓CNE2细胞侵袭能力（P〈0.01）。结论 miR-34a可抑制鼻咽癌CNE2细胞生长与侵袭。     

MCL1在鼻咽癌中的表达水平及临床意义

目的 研究髓样细胞白血病蛋白1（myeloid cell leukemia-1 protein,MCL1）在鼻咽癌组织和鼻咽癌细胞中的表达情况,并分析其表达水平与鼻咽癌患者临床预后指标之间的关系。方法 运用免疫组化方法检测鼻咽癌组织和正常鼻腔黏膜组织中MCL1蛋白的表达差异,并分析MCL1表达与鼻咽癌患者临床病理指标之间的关系;运用Western blot法检测人鼻咽癌细胞系CNE2、CNE1以及人永生化鼻黏膜上皮细胞NP-69中MUC1蛋白表达情况。结果 免疫组化结果显示,鼻咽癌组织中MCL1表达阳性率为58.3%（35/60）,在正常鼻腔黏膜组织中MCL1阳性表达率23.8%（5/21）,鼻咽癌组织的MCL1表达水平显著高于正常鼻腔黏膜组织（P<0.05）;MCL1的表达水平与鼻咽癌患者的T分级、临床分期及淋巴结转移显著相关（P<0.05）,而与患者年龄、性别及分化程度无显著相关性（P>0.05）;同时,MCL1在鼻咽癌细胞系CNE2及CNE1中的表达高于NP69细胞,差异均有统计学意义（P<0.05）;结论 MCL1在鼻咽癌中可能起到促进肿瘤进展的作用,患者的MCL1表达水平可能与其预后相关。     

microRNA-372靶向SDC1抑制人胰腺癌上皮间质转化及侵袭

目的 探讨microRNA-372（miR-372）靶向调控SDC1基因表达抑制人胰腺癌上皮间质转化及侵袭的机制研究。方法 收集人胰腺癌及癌旁组织手术标本;qRT-PCR和免疫组化法检测组织中miR-372和SDC1表达;筛选miR-372表达水平最高的人胰腺癌细胞系进行分组和转染,实验分成6组,即空白对照组、阴性对照组、miR-372 mimic组、miR-372 inhibitor组、siRNA-SDC1组和miR-372 inhibitor+siRNA-SDC1组;双荧光素酶报告基因试验验证miR-372和SDC1基因的靶向关系;CCK8法、流式细胞术、划痕试验及Transwell试验分别检测各组细胞转染后增殖能力、凋亡情况、迁移及侵袭能力;qRT-PCR和Western blot法检测各组细胞转染后SDC1和上皮细胞标志物E-cadherin及间质表型细胞标志物Vimentin的表达水平。结果 SDC1蛋白在胰腺癌组织中的阳性表达率明显高于癌旁正常组织（P<0.05）。胰腺癌中miR-372、SDC1表达与肿瘤分化程度、转移和TNM分期显著相关（P<0.05）。双荧光素酶报告基因试验结果显示SDC1是miR-372的靶基因。与空白对照组和阴性对照组比较,miR-372 mimic组和siRNA-SDC1组细胞的生长、迁移和侵袭受到抑制,凋亡率增加,SDC1和Vimentin的表达水平明显降低,E-cadherin的表达水平明显升高（P均<0.05）;而miR-372 inhibitor组的趋势与miR-372 mimic组和siRNA-SDC1相反（P均<0.05）;然而,miR-372 inhibitor+siRNA-SDC1组与空白对照组和阴性对照组比较,各项指标差异均无统计学意义。结论 miR-372可能通过靶向抑制SDC1基因表达,抑制人胰腺癌细胞的迁移、侵袭和上皮间质转化。     

miR-577在胃癌中的表达及对SGC-7901细胞侵袭的影响

目的 研究微小RNA 577（microRNA-577,miR-577）在人胃癌（gastric cancer,GC）组织内的表达情况及对肿瘤细胞侵袭能力的影响。方法 选择2013年1月~2015年3月于第三军医大学大坪医院普通外科收治并行手术切除的70例胃癌与对应癌旁组织。通过qRT-PCR方法检测miR-577在组织中的相对表达量,分析miR-577的临床病理意义;采用人工合成的miR-577模拟物（miR-577 mimics）转染人胃癌SGC-7901细胞,并检测SGC-7901细胞过表达miR-577后侵袭能力及miR-577下游潜在靶点β-catenin mRNA及蛋白的表达变化。结果 MiR-577在胃癌组织中表达显著下调（P=0.000）并与与淋巴结转移（P=0.003）、高TNM分期（P=0.006）等不良临床特征呈正相关;过表达miR-577能够显著抑制SGC-7901细胞侵袭能力（P=0.000）及细胞内β-catenin的表达水平。结论 miR-577在胃癌中低表达,并可能通过下调β-catenin的表达来抑制胃癌细胞的侵袭。     

miRNA-135b 对鼻咽癌细胞侵袭 和转移的促进作用

目的 研究miRNA-135b 对鼻咽癌CNE1 细胞系侵袭和转移的影响和作用机制。方法 用逆转录聚 合酶链反应（RT-PCR）检测miRNA-135b 在鼻咽癌CNE1 细胞系中的表达,分别在鼻咽癌CNE1 细胞系中 过表达和下调miRNA-135b,通过微孔滤膜培养小室及双室联合培养系统（Transwell TM）检测肿瘤细胞迁移和 侵袭能力,采用Western blotting 检测大型肿瘤抑制因子同源蛋白2（LATS2）的表达。结果 miRNA-135b 在 鼻咽癌CNE1 细胞系中高表达,通过增强miRNA-135b 表达,鼻咽癌CNE1 细胞的侵袭和转移能力有所提升; 通过靶向miRNA-135b 干扰载体,下调miRNA-135b 表达,鼻咽癌CNE1 细胞的侵袭和转移能力有所下降。 增强miRNA-135b 表达,LATS2 活性和表达下降,而下调miRNA-135b 表达,LATS2 活性和表达上升。结 论 miRNA-135b 过表达会提升鼻咽癌CNE1 细胞的侵袭和转移能力,下调miRNA-135b 表达可有效抑制鼻 咽癌细胞增殖,促进细胞凋亡。miRNA-135b 的表达可能通过影响LATS2 活性影响鼻咽癌细胞的生长。     

长链非编码ATB通过抑制miR-200c的表达促进乳腺癌细胞的侵袭转移

目的 探讨长链非编码ATB通过抑制miR-200c的表达促进乳腺癌细胞侵袭转移的作用及其机制.方法 qPCR检测不同乳腺癌细胞株中ATB和miR-200c的表达情况;双荧光素酶报告基因检测ATB与miR-200c的相互作用;MTT细胞增殖实验和Transwell侵袭实验检测沉抑制ATB后乳腺癌细胞增殖和侵袭能力的变化;MTT细胞增殖实验和Transwell侵袭实验检测抑制ATB后沉默miR-200c对乳腺癌细胞增殖和侵袭能力的逆转作用;Western blot检测抑制ATB后ZEB1和ZNF217蛋白的表达.结果 与其他乳腺癌细胞相比,SKBr-3细胞株ATB表达水平最低,miR-200c的表达水平最高;ATB能与miR-200c的位点特异性结合,调控其表达活性;抑制ATB后可以增强乳腺癌细胞的增殖和侵袭能力,而沉默miR-200c后可以在一定程度上逆转ATB对乳腺癌细胞增殖和侵袭能力的影响;抑制ATB后ZEB1和ZNF217蛋白表达水平得到一定的恢复.结论 ATB在乳腺癌发生、发展过程中起重要作用,ATB可以靶向调节miR-200c调控乳腺癌细胞的增殖和侵袭能力.     

circ_0000069靶向miR-338-3p对非小细胞肺癌A549细胞增殖、迁移和侵袭的影响

目的探讨环状RNA 0000069(circ_0000069)是否靶向miR-338-3p调控非小细胞肺癌(NSCLC)A549细胞增殖、迁移和侵袭。 方法采用qRT-PCR技术检测NSCLC患者癌组织、癌旁正常组织中circ_0000069和miR-338-3p表达量。采用CCK-8、Transwell实验分析抑制circ_0000069或过表达miR-338-3p对A549细胞增殖活力、迁移和侵袭的影响。使用荧光素酶酶报告实验和qRT-PCR分析circ_0000069和miR-338-3p之间的相互作用。 结果NSCLC组织中circ_0000069表达量较癌旁组织升高(P＜0.05),而miR-338-3p表达量显著降低(P＜0.05)。抑制circ_0000069或过表达miR-338-3p后A549细胞活力、迁移和侵袭细胞数显著降低(P＜0.05)。miR-338-3p是circ_0000069的靶基因,circ_0000069靶向负调控miR-338-3p表达。干扰miR-338-3p表达显著减弱circ_0000069对A549细胞增殖、迁移和侵袭的抑制(P＜0.05)。 结论抑制circ_0000069通过上调miR-338-3p抑制NSCLC细胞A549增殖、迁移和侵袭。     

miR-138在鼻咽癌患者中的预后价值及其对细胞增殖、迁移和侵袭的影响

目的 探讨miR-138在鼻咽癌中的临床意义以及对癌细胞增殖、迁移和侵袭的影响。 方法 采用实时荧光定量PCR检测鼻咽癌组织和癌细胞CNE1中miR-138的表达量。采用Kaplan-Meier生存分析和Cox回归分析评估miR-138的预后价值。构建体外调控miR-138表达量的鼻咽癌细胞体系,采用MTT法和Transwell实验法分析miR-138对癌细胞CNE1生命活动的影响。 结果 miR-138在鼻咽癌组织和细胞中的表达量均显著下调(P<0.001),且与癌症患者的淋巴结转移(P=0.007)及TNM分期(P=0.003)密切相关。生存曲线表明,低表达miR-138的鼻咽癌患者较高表达患者具有较差的总生存率(P=0.002),且可以作为一个独立的预后因子(HR=2.603,95%CI=1.245～5.441,P=0.011)应用在鼻咽癌的治疗中。体外细胞实验证明,高表达miR-138具有抑制鼻咽癌细胞CNE1增殖、迁移和侵袭的能力(P<0.05),而抑制miR-138的表达则显著促进癌细胞的各种生命活动(P<0.05)。 结论 miR-138与鼻咽癌的预后密切相关,且可以明显抑制鼻咽癌细胞CNE1的增殖、迁移和侵袭能力。     

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.