通过下调FOXM1的新型小分子抑制剂1-3-51抗胃癌细胞的作用

目的 研究新型小分子抑制剂1-3-51在胃癌中调控FOXM1 (Forkhead box protein M1)表达及其抗癌的作用.方法 CCK-8实验检测1-3-51处理后的BGC823和MKN45细胞IC50;Western blot检测胃癌细胞FOXM1表达;qRT-PCR检测胃癌细胞FOXM1及其下游分子Cyclin D1和MMP9 mRNA表达;流式细胞术测定细胞周期情况;Transwell实验检测胃癌细胞迁移和侵袭能力;裸鼠皮下移植瘤模型检测1-3-51抗胃癌增殖效果;在胃癌细胞BGC823中转染FOXM1过表达质粒及其空载质粒,CCK-8实验、Transwell实验分别检测1-3-51对胃癌细胞活性、迁移及侵袭等的影响.结果 新型小分子抑制剂1-3-51在BGC823和MKN45细胞作用48 h后的IC50分别为(5.429±0.225) μmol/L、(5.169±0.239) μmol/L,并可以显著抑制FOXM1的表达及其下游分子Cyclin D1和MMP9 mRNA表达.1-3-51显著抑制胃癌细胞的增殖、侵袭及迁移.过表达FOXM1后,可部分逆转其抑制细胞增殖、侵袭及迁移能力.结论 小分子抑制剂1-3-51在胃癌细胞中可通过抑制FOXM1及其下游分子Cyclin D1和MMP9等的表达,进而降低胃癌细胞增殖、迁移、侵袭能力.     

EGCG经PI_3K-AKT信号通路诱导人胃癌BGC-823细胞G1期阻滞

目的探讨表没食子儿茶素没食子酸酯（Epigallocatechin-3-gallate，EGCG）通过PI3K-AKT通路对人胃癌BGC-823细胞周期调控的分子机制。方法采用噻唑蓝比色法（MTT）检测BGC-823细胞的存活率；碘化丙啶（PI）染色流式细胞术（flow cytometry，VCM）检测EGCG处理前后BGC823细胞周期的变化；Western blot检测PI3K-AKT信号传导通路相关蛋白的表达。结果EGCG能够抑制BGC823细胞的生长，且这种抑制作用与诱导BGC823细胞G1期阻滞有关。EGCG可以下调巩K-AKT信号通路蛋白Akt的磷酸化表达，对通路下游周期相关蛋白cyclinD1的表达也有抑制作用。结论EGCG能够通过活化PLK-AKT信号通路发挥诱导人胃癌BGC823细胞G1期阻滞的作用。     

白藜芦醇诱导胃癌BGC⁃823细胞凋亡的分子机制

目的：探究白藜芦醇对人胃癌细胞BGC823的作用及其分子机制。方法：采用MTT法检测白藜芦醇对BGC823细胞增殖的影响;流式细胞术测定白藜芦醇对BGC823细胞凋亡、活性氧水平和线粒体膜电位的影响;Western blot法检测白藜芦醇对BGC823细胞cleaved caspase?3和cleaved caspase?9蛋白表达的影响。结果：MTT结果显示白藜芦醇对胃癌BGC823细胞具有明显的增殖抑制作用,且呈时间和剂量依赖关系;31.25~500.00 μmol/L白藜芦醇作用24 h和48 h可诱导BGC823细胞凋亡,500 μmol/L作用24 h和48 h,细胞凋亡率分别达（40.42±2.64）%和（75.02±2.36）%。流式细胞术结果显示白藜芦醇作用24 h后可使BGC823细胞内活性氧（reactive oxygen species,ROS）增多;与对照组相比,62.50~500.00 μmol/L白藜芦醇作用细胞24 h后,细胞出现了显著的线粒体膜电位降低（P < 0.05）;Western blot结果显示白藜芦醇以浓度依赖方式上调cleaved caspase?9和cleaved caspase?3的表达。结论：白藜芦醇可以显著诱导细胞凋亡,其诱导细胞凋亡与ROS增多、线粒体膜电位降低和调节cleaved caspase?9和cleaved caspase?3的表达有关。     

TRAF1基因干扰质粒的构建及其对胃癌细胞生物学功能的影响

目的:构建T F1基因干扰重组质粒及建立T F1基因干扰瞬时转染胃癌细胞模型,然后通过干扰目标胃癌细胞中的T F1基因来检测T F1对胃癌细胞的生物学功能的影响。方法:首先通过real-time PCR和Western印迹验证T F1在胃癌细胞系BGC823,SGC7901,MGC803中的表达,筛选出T F1表达量最高的一株细胞作为干扰转染的目标细胞;设计及构建3个T F1的shRNA表达载体pLVX-shRNA-T F1-shRNA1~3,将其分别转染至目标胃癌细胞系中,应用real-time PCR和Western印迹筛选出干扰效率最强的shRNA,最后通过M及流式细胞术检测T F1对胃癌细胞凋亡能力的影响,通过Transwell小室迁移实验来检测对其迁移功能的影响。结果:与GES-1比较,T F1基因在BGC823,SGC7901,MGC803中的表达均有上调(P<0.05),其中以BGC823表达最高;3个T F1shRNA对T F1基因均有抑制作用,以pLVX-shRNA-T F1-shRNA2干扰效果最强;干扰胃癌细胞BGC823中的T F1可使细胞生长活力减弱,凋亡明显增加,但对其迁移功能无明显影响。结论:T F1基因在胃癌细胞系BGC823中上调最明显;pLVX-shRNA-T F1-shRNA2可成功干扰BGC823中T F1的表达;T F1可抑制BGC823细胞的凋亡。     

下调STYK1基因表达可抑制结直肠癌细胞生长与侵袭

目的 探讨STYK1基因下调对结直肠癌细胞HCT-15增殖、凋亡和迁移以及MEK/ERK和PI3 K/AKT信号通路的影响.方法 采用基因干扰技术抑制结直肠癌细胞HCT-15中STYK1的表达,Western blot检测抑制效果,CCK-8检测STYK1表达下调对HC-T15细胞增殖的影响,流式细胞仪检测STYK1表达下调对HC-T15细胞凋亡的影响,Transwell小室实验检测STYK1表达下调对HCT-15细胞侵袭的影响,通过Western blot检测STYK1表达下调对HCT-15细胞MEK/ERK和PI3 K/AKT信号通路活性的影响.结果 与干扰对照组相比,STYK1表达下调明显抑制HCT-15细胞增殖活性(P＜0.01)和侵袭能力(P＜0.01),并诱导细胞凋亡(P＜0.01);同时,STYK1表达下调明显抑制HCT-15细胞ERK1/2和AKT蛋白的磷酸水平.结论 STYK1基因干扰可以抑制结直肠癌细胞的增殖和侵袭,并诱导凋亡,其机制可能与抑制MEK/ERK和PI3K/AKT信号通路活性有关.     

OPG/RANKL/RANK系统参与牙槽骨吸收及重建过程作用初探

目的 初步探讨护骨素OPG、核因子κB受体活化因子配体RANKL、核因子κB活化因子RANK在牙周炎牙槽骨吸收及骨重建过程中的作用.方法 对小鼠单核巨噬细胞白血病细胞株RAW264.7实验组进行体外诱导培养,对照组为完全培养基培养;小鼠成骨样细胞株MC3T3-E1实验组采用前期实验收集的骨吸收上清处理,对照组采用完全培养基培养;以免疫荧光等方法检测细胞OPG、RANKL、RANK的表达.30只8周龄雄性SD大鼠建立实验性牙槽骨吸收模型,研究其吸收重建规律.以免疫组化S-P法检测OPG、RANKL、RANK的表达情况.结果 实验组MC3T3-E细胞经骨吸收上清处理7d后,OPG蛋白平均荧光强度为(29.636±5.652),对照组为(15.568±1.229),表达显著上调(P＜0.01);但实验组RANKL蛋白平均荧光强度为(6.806±1.738),对照组为(18.082±2.732),表达被显著抑制(P＜0.01),OPG/RANKL比值在上清处理后高于对照组(P＜0.05).采用大肠杆菌内毒素E-LPS注射法成功制备大鼠牙槽骨吸收模型.在牙槽骨吸收区域OPG水平较无骨吸收区域有所降低,RANKL的表达则相反.结论 OPG、RANKL、RANK参与了大鼠实验性骨吸收活动;破骨细胞骨吸收上清可能通过改变OPG与RANKL的比值,从而影响骨重建中骨形成与骨吸收的动态平衡.     

SP-141抑制胃癌细胞增殖、迁移并促进凋亡

目的：探讨MDM2的新型小分子抑制剂SP-141对胃癌细胞株MGC803和BGC823增殖、凋亡以及迁移的影响。方法：胃癌细胞经SP-141处理后,采用CCK-8法、克隆形成实验、流式细胞术、Hoechst染色法和划痕实验分别检测细胞存活率、细胞增殖、周期分布、凋亡以及迁移能力改变。Western blot检测相关分子蛋白质表达水平。结果：TCGA数据库中32对胃癌组织中的MDM2 mRNA表达水平高于癌旁组织（P<0.01）;5种胃癌细胞株均检出MDM2,表达水平差异不大;SP-141抑制MGC803和BGC823的细胞存活率以及克隆形成,诱导其细胞周期阻滞在G2/M期;SP-141增加细胞凋亡发生率,并下调Bcl-2同时上调Bax、Caspase-3剪切体、PARP剪切体的蛋白表达;SP-141抑制细胞迁移并伴有FAK蛋白表达量的下降。结论：MDM2的新型小分子抑制剂SP-141可有效抑制胃癌细胞增殖、迁移并促进细胞凋亡。     

全反式视黄酸衍生物逆转高水平胰岛素对胃癌细胞BGC-823迁移的作用研究

目的 探讨高水平胰岛素条件下全反式视黄酸衍生物4-氨基-2-三氟甲基-苯基-维甲酸酯(ATPR)对人胃癌细胞BGC-823迁移能力的影响及其潜在的分子机制.方法 首先采用不同浓度胰岛素刺激BGC-823细胞,CCK-8法检测胰岛素对细胞增殖能力的影响;然后采用等同人高胰岛素血症水平(≥0.5 ng/ml)而对BGC-823增殖能力没有影响的胰岛素剂量(＜ 10.0 ng/ml),联合ATPR处理BGC-823细胞后,通过细胞划痕实验和Transwell实验明确BGC-823细胞迁移能力的改变;Western blot检测与细胞迁移能力相关蛋白的表达水平.结果 CCK-8结果显示,与对照组比较,0.5、5.0 ng/ml胰岛素作用48 h后对BGC-823细胞的增殖无影响,而10.0 ng/ml胰岛素能够促进BGC-823细胞增殖;细胞划痕和Transwell实验显示,5.0 ng/ml胰岛素明显促进BGC-823细胞的迁移能力;但是,联合ATPR后能够抑制胰岛素对BGC-823迁移的促进作用,作用与肌球蛋白轻链激酶(MLCK)抑制剂ML-7类似;Western blot结果显示胰岛素促进MLCK蛋白表达且下调胃特异性紧密连接蛋白Claudin 18的表达,而ATPR联合处理后能抑制MLCK并促进Claudin 18蛋白的表达.结论 ATPR可逆转高水平胰岛素对胃癌细胞BGC-823迁移的促进作用,可能与其抑制胰岛素对MLCK和Claudin 18的调控密切相关.     

PI3''''K/AKT通路抑制剂对人胃癌细胞Caspase-9表达的影响

目的:检测PI3'K/Akt通路抑制剂对抗肿瘤药物诱导人胃癌细胞凋亡过程的影响.方法:将胃癌细胞系BGC-823细胞分为6组:空白对照组、渥曼青霉素组、足叶乙甙组、渥曼青霉素 足叶乙甙组、阿霉素组和渥曼青霉素 阿霉素组.按照上述分组用药物干预BGC-823细胞24 h,采用NaI法提取各组细胞的DNA,Western blot检测caspase-9表达变化及用琼脂糖凝胶电泳检测细胞凋亡情况.结果:① DNA琼脂糖凝胶电泳示空白对照组未见DNA 梯状谱,其他5组均出现明显的DNA梯状谱;② Western blot 结果提示阿霉素与足叶乙甙干预后,BGC-823的Caspase-9活性与未处理组相比增强,加用渥曼青霉素处理后,BGC-823的Caspase-9活性与未处理组相比明显增强,与单用阿霉素或足叶乙甙处理组相比有所增强.结论:PI3K/AKT通路抑制剂能促进胃癌细胞凋亡,增强抗肿瘤化疗药物的疗效 .     

AMPK对高糖环境下人牙周膜细胞中RANKL表达的调节作用

目的：高糖可在一定程度上影响牙周膜细胞的生物学功能。本研究皆在探讨高糖对人牙周膜细胞中RANKL表达的影响及该影响是否可被AMPK活化所调节 方法：Real-time PCR和Western blot方法检测细胞中RANKL和 OPG表达,并通过检测AMPK活化特异性底物SAMS及Westernblot检测AMPK磷酸化,观察细胞中AMPK活性变化。此外,通过通过Real-time PCR和Western blot方法检测AMPK活化对牙周膜细胞中RANKL表达的影响 结果：高糖作用下人牙周膜细胞中RANKL mRNA和蛋白表达增高,且AMPK活性明显下降;Compound C可通过抑制AMPK活性上调RANKL表达,而二甲双胍可通过激活AMPK下调RANKL表达;另外,我们还发现二甲双胍可通过激活AMPK显著减少高糖环境下牙周膜细胞中RANKL表达 结论：结果提示高糖作用下牙周膜细胞中RANKL表达升高与AMPK的磷酸化及活性受到抑制有关,且AMPK活化可参与调节牙周膜细胞中RANKL表达     

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…     

Dr.Zhang Ren''''s Experience in the Acupuncture Treatment of Different Diseases with the Same Therapeutic Principle

Dr.Zhang Ren,the chief physician,is the chairman of Shanghai Acupuncture and Moxibustion Association.Having been engaged in medicine for about 40 years,he is experienced in treating various intractable diseases.In his long years of clinical practice,he advocates taking the TCM differentiation as the basis to seek for the acupuncture method for treatment of modern intractable diseases.The author of this essay had the fortune to follow Dr.Zhang in study.The following is a summary of Dr.Zhang's experience in the acupuncture treatment for different intractable diseases with the same therapeutic principle.     

Luo Lingjie''''s Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.     

猪肺磷脂注射液联合经鼻持续气道正压通气对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性的影响

目的 探讨猪肺磷脂注射液联合经鼻持续气道正压通气(NCPAP)对呼吸衰竭早产儿的临床疗效及肌酸激酶同工酶活性(CK-MB)的影响.方法 选取呼吸衰竭早产儿80例,分为观察组和对照组各40例.对照组采用NCPAP给氧治疗,观察组给予NCPAP给氧联合猪肺磷脂气管内给药.观察两组患儿治疗前及治疗12h、24 h后PaO2、PaCO2、血氧饱和度(SaO2)、pH的变化情况,检测治疗前及治疗5d后血清CK-MB水平;评估两组患儿的临床治疗效果.结果 两组患儿PaO2、PaCO2、SaO2、pH比较,差异均有统计学意义(P＜0.05),其中观察组治疗后的PaO2、SaO2、pH均高于对照组,PaCO2则低于对照组.两组的PaO2、SaO2、pH均随观察时间延长而升高(P＜0.05),PaCO2均随观察时间的延长而降低(P＜0.05).观察组治疗有效率为87.5％,显著高于对照组的70.0％ (P ＜0.05).治疗5d后两组患儿血清CK-MB水平均较前降低(P＜0.05),且观察组明显低于对照组(P＜0.05).结论 猪肺磷脂注射液气管内给药联合NCPAP可以显著降低呼吸衰竭早产儿CK-MB的含量,提高治疗有效率,起到很好的呼吸循环支持作用.     

Survey and practice of reporting quality of randomized controlled clinical trials on traditional Chinese medicine

Evidence obtained from randomized controlled trials （RCTs） has been generally accepted as the gold standard in the evaluation of clinical effectiveness. Readers need to understand the trial design, implementation, results, analysis and interpretation, so as to fully Jnderstand the results of RCTs. Thus, the investigators of RCTs have to report these items in a complete, accurate and clear manner. Since 1998, we have conducted several evaluations on the reporting quality of RCTs published in Chinese journals on traditional Chinese medicine （TCM） and results have shown that there is an urgent need for higher quality RCTs on TCM.     

TCM Treatment of Ankylosing Spondylitis by Tonifying the Kidney and Strengthening the Governor Vessel

Ankylosing spondylitis is a chronic and progressive disorder with inflammation mainly involving the central axis joints. It mainly affects the cervical spine and the lumbosacral area, with the pathogenesis closely related to the kidney and the Governor Vessel (GV). TCM holds that the syndrome is deficiency in origin and excess in superficiality, which is due to insufficiency of the kidney, deficiency of GV, and blocking of the channels with the invasion of exogenous evil, leading to poor circulation of qi and blood and malnutrition of the bones, muscles and joints. The TCM method of tonifying the kidney and strengthening GV to regulate circulation of qi and blood and check the arthralgia pain should be adopted, with the Kidney-Tonifying and GV Strengthening Decoction (益肾强督汤) prescribed.     

IMPROVING P-gp EXPRESSION IN HUMAN MONONUCLEAR CELLS IN VITRO TRANSFECTED BY MULTIDRUG RESISTANCE-1 mRNA

CHEMOTHERAPY playsa greatrolein the treat- ment of malignanttumors,especiallyingynecolo- gicalones.But inanticancerchemotherapy,leuko-cytopeniaisfrequentlytheprimarydose-limitingsideeffect factor.Moreover,cancersarefrequentlychemoresistantbe-causeof overexpressionof P-glycoprotein(P-gp), which isencodedby multidrugresistancegene (MDR1 ) and detectableinup to50% ofhuman cancersand renderscellsresistancetoanticancerdrugs.The safetyand potentialtherapeuticbenefitof mdr1 gene transferredto h…