Rac1在肝癌细胞株生长和侵袭中的意义

目的:证实Rac1在肝癌细胞的生长与侵袭中的意义,寻找可能的肿瘤基因治疗新靶点。方法:特异性抑制Rac1蛋白表达的siRNA干扰肝癌高转移细胞株97-H,观察肝癌高转移细胞株97-H内Rac1表达、细胞生长、肿瘤细胞的活力和体外侵袭力的变化。结果:肝癌高转移细胞株97-H内Rac1表达、细胞生长受抑制,肿瘤细胞的活力和体外侵袭力显著降低。结论:Rac1表达的减少能够显著抑制肝癌细胞的生长和侵袭,可能成为肿瘤基因治疗新的靶点。     

Effects of vitamin D analogues EB1089 on proliferation,apoptosis and invasion of heptocellular cells

目的 探讨维生素D类似物EB1089对肝癌细胞株增殖、凋亡和侵袭的影响.方法 体外培养肝癌细胞株MHCC97H,用不同浓度的EB1089分别培养不同的时间段.用四唑蓝比色实验(MTT)检测细胞的存活和生长,用流式细胞仪检测细胞的凋亡,用Transwell和划痕实验检测细胞的侵袭迁移能力,用RT-PCR方法检测雌激素受体α(ERα)、核转录因子p65(NF-κB p65)和基质金属蛋白酶-9(MMP-9)的mRNA的表达.结果 EB1089可以抑制肝癌细胞株的增殖,诱导肝癌细胞发生凋亡,抑制肝癌细胞的侵袭迁移.同时,EB1089增加MHCC97H细胞中ERα mRNA表达,下调NF-κB p65和 MMP-9 mRNA表达.结论 EB1089可能通过ERα途径抑制肝癌细胞株的增殖、侵袭迁移能力,诱导其凋亡.     

JWA抑制人肝癌细胞的黏附和迁移侵袭潜能

探讨JWA蛋白在人肝癌细胞黏附和迁移侵袭过程中的作用?方法:应用小干扰RNA下调低转移潜能人肝癌细胞株MHCC97L中JWA蛋白表达水平,MTT法检测MHCC97L增殖;应用细胞小室检测MHCC97L在JWA不同蛋白水平时的迁移及侵袭潜能变化;黏附试验检测MHCC97L细胞黏附能力变化?结果:研究发现人肝癌细胞株MHCC97L中的JWA表达水平下降后,肝癌细胞迁移及侵袭能力明显增强,细胞对细胞外基质纤维连接蛋白的黏附力增加?结论:RNA干扰JWA引起肝癌细胞的JWA蛋白表达水平下降,导致肝癌细胞的黏附?迁移及侵袭能力增强,结果提示JWA可能抑制人肝癌细胞侵袭和转移?     

CXCR7/CXCL12轴对人肝癌细胞增殖能力的影响

目的观察不同恶性程度的肝癌细胞株中趋化因子受体CXCR7的表达情况,探讨CXCL12/CXCR7生物学轴对人肝癌细胞增殖能力的影响。方法以肝癌成瘤高转移细胞株MHCC97-H及成瘤低转移细胞株PLC/PRF/5为研究对象。采用RT-PCR、Western blot免疫印迹法检测两株肝癌细胞的CXCR7 mRNA与蛋白表达情况,应用CXCR7激动剂(重组人CXCL12)和特异拮抗剂(CCX771)分别刺激和阻断CXCR7功能,应用Alamarblue法检测细胞的增殖能力。结果 MHCC97-H细胞株CXCR7的mRNA与蛋白表达明显高于PLC/PRF/5细胞株,CXCL12能明显增强MHCC97-H细胞增殖能力(P<0.01),CXCR7阻断剂CCX771作用后,MHCC97-H细胞增殖能力明显下降(P<0.01);CXCL12对PLC/PRF/5细胞增殖能力无明显影响(P>0.05)。结论 CXCL12/CXCR7生物学轴与肝癌细胞株增殖能力有一定的关系,CXCR7拮抗剂CCX771可明显抑制肝癌细胞的增殖能力。     

C×CR7／CXCLl2轴对人肝癌细胞增殖能力的影响

目的观察不同恶性程度的肝癌细胞株中趋化因子受体CXCR7的表达情况,探讨CXCLl2／CXCR7生物学轴对人肝癌细胞增殖能力的影响。方法以肝癌成瘤高转移细胞株MHCC97-H及成瘤低转移细胞株PLC／PRF／5为研究对象。采用RT-PCR、Westernblot免疫印迹法检测两株肝癌细胞的CXCR7mRNA与蛋白表达情况,应用CXCR7激动剂（重组人CXCLl2）和特异拮抗剂（CCX771）分别刺激和阻断CXCR7功能,应用Alamarblue法检测细胞的增殖能力。结果MHCC97-H细胞株CXCR7的mRNA与蛋白表达明显高于PLC／PRF／5细胞株,CXCLl2能明显增强MHCC97-H细胞增殖能力（P〈0．01）,CXCR7阻断剂CCX771作用后,MHCC97一H细胞增殖能力明显下降（P〈0．01）;CXCLl2对PLC／PRF／5细胞增殖能力无明显影响（P〉0．05）。结论CxCLl2／CXCR7生物学轴与肝癌细胞株增殖能力有一定的关系,CXCR7拮抗剂CCX771可明显抑制肝癌细胞的增殖能力。     

RNA干扰HBx基因对肝癌细胞化疗效果的影响

目的：探讨RNA干扰技术和传统的化疗方法联合应用对肝癌细胞生长抑制作用的影响,并选择高效的治疗组合,研究其凋亡机制。方法：鉴定3组肝癌细胞——MHCC97-H(原肝癌细胞株),HK3细胞（转染空质粒的MHCC97-H细胞）和21543细胞（转染含HBx-shRNA质粒的MHCC97-H细胞）;RT-PCR检测RNA干扰对HBx mRNA沉寂作用,CCK8和TUNEL分别检测RNA干扰HBx后化疗药物对MHCC97-H肿瘤细胞生长的抑制及诱导细胞凋亡情况。结果：RT-PCR结果显示,与MHCC97-H细胞组比较,21543细胞的HBx mRNA水平下降约91%,HK3细胞HBx mRNA水平下降不明显;RNA靶向干扰HBx基因后的肝癌细胞（21543细胞）较原肝癌细胞（MHCC97-H细胞）和HK3细胞增殖明显减慢,而后两种细胞差别无统计学意义;3种不同细胞加用3种不同浓度的氟尿嘧啶(0~120 mg/L)、顺铂(0~32 mg/L)后细胞生长明显减慢并呈浓度依赖性,以RNA靶向干扰HBx基因后的21543细胞生长抑制最明显;相同浓度的氟尿嘧啶对3组不同肝癌细胞均可引起细胞凋亡,以21543细胞凋亡最明显。结论：RNA干扰HBx可明显抑制肝癌细胞的增殖,并增加肝癌细胞对化疗药物的敏感性;RNA干扰HBx基因和化疗药物联合应用使肝癌细胞凋亡更明显,细胞增殖速度明显减慢。     

维生素D类似物EB1089对肝癌细胞增殖凋亡侵袭的影响

目的探讨维生素D类似物EB1089对肝癌细胞株增殖、凋亡和侵袭的影响。方法体外培养肝癌细胞株MH-CC97H,用不同浓度的EB1089分别培养不同的时间段。用四唑蓝比色实验(MTT)检测细胞的存活和生长,用流式细胞仪检测细胞的凋亡,用Transwell和划痕实验检测细胞的侵袭迁移能力,用RT-PCR方法检测雌激素受体α(ERα)、核转录因子p65(NF-κB p65)和基质金属蛋白酶-9(MMP-9)的mRNA的表达。结果 EB1089可以抑制肝癌细胞株的增殖,诱导肝癌细胞发生凋亡,抑制肝癌细胞的侵袭迁移。同时,EB1089增加MHCC97H细胞中ERαmRNA表达,下调NF-κB p65和MMP-9 mRNA表达。结论 EB1089可能通过ERα途径抑制肝癌细胞株的增殖、侵袭迁移能力,诱导其凋亡。     

β细胞素下调对肝癌细胞增殖能力的影响及其机制探讨

目的:在肝癌细胞系中通过下调β细胞素(BTC)的表达研究其对肝癌细胞侵袭能力的影响并探究其影响机制。方法:通过siRNA转染下调BTC后,通过Transwell侵袭实验检测SMMC7721和MHCC97-H细胞的侵袭能力变化,并用RT-PCR和Western blot检测与肝癌侵袭能力相关的PI3K/AKT-XIAP信号通路的表达变化。结果:与阴性对照组相比,siRNA下调组中SMMC7721和MHCC97-H细胞的侵袭能力明显降低,PI3K/AKT-XIAP信号通路受到明显抑制。结论:下调BTC表达可能通过抑制PI3K/AKT-XIAP信号通路抑制肝癌细胞的侵袭能力。     

KAI1基因对肝癌细胞侵袭、转移的影响及其分子机制

目的研究肿瘤转移抑制基因KAI1对肝癌细胞侵袭、转移的影响及其分子机制,为肝癌的基因治疗提供理论和实验依据.方法用亚克隆技术构建KAI1基因正、反义真核表达质粒,DOTAP脂质体法将其分别转入高转移潜能的人MHCC97-H肝癌细胞,以限制性酶切、PCR、Western blot及免疫组化鉴定基因重组及转染是否成功.电镜观察细胞超微形态变化,流式细胞仪检测细胞周期分布,细胞生长曲线和平板法集落形成试验观察细胞生长和集落形成能力,Boyden Chamber试验观察细胞侵袭能力,微管吸吮技术测定细胞的粘弹性和粘附力.BALB/CA 4～6周龄雄性裸鼠80只,随机分正义组、反义组、亲本细胞组及空载体组进行皮下(n=40)和原位肝(n=40)种植试验,观察体内的成瘤性、侵袭性及远处转移能力,免疫组化研究癌转移灶AFP和肿瘤组织细胞外基质(ECM)及粘附分子的表达状况.结果(1)成功构建了KAI1正、反义基因真核表达质粒,并将其分别转入MHCC97-H人肝癌细胞;(2)发现转入KAI1基因后可导致肝癌MHCC97-H细胞的一些细胞器数量和形态的变化;(3)KAI1基因对肝癌MHCC97-H细胞的体内外生长、细胞周期及体内成瘤性无明显影响;(4)反义KAI1基因可使MHCC97-H细胞的克隆形成能力增强、对体内外肝癌细胞的侵袭和转移有促进作用;(5)KAI1基因表达上调能使MHCC97-H细胞的体内外侵袭能力降低;(6)KAI1基因抑制肝癌侵袭和转移的机制可能与细胞粘弹性增加、ECM表达上调、粘附分子表达下调及粘附力下降等变化有关.结论KAI1基因在肝癌的侵袭转移过程中发挥着重要作用,上调KAI1基因表达能明显降低肝癌细胞的侵袭和转移能力.     

GPC3对肝癌细胞株MHCC97-L增殖的影响

目的通过构建磷脂酰肌醇蛋白聚糖3(GPC3)增强型绿色荧光蛋白真核表达载体,研究GPC3对人肝癌细胞株MHCC97-L增殖的影响。方法用基因重组及限制性内切酶技术构建并鉴定表达载体pEGFP-c3-GPC3,通过脂质体介导转染人肝癌细胞株MHCC-97L,流式细胞术和G418筛选建立稳定细胞株;采用荧光定量PCR、Western blotting分别检测MHCC-97L中GPC3 mRNA和蛋白的表达情况,并在荧光显微镜下观察细胞荧光;MTT法检测GPC3对肝癌细胞MHCC97-L体外增殖能力的影响。结果经酶切及测序鉴定,确认成功构建真核表达载体pEGFP-c3-GPC3,荧光显微镜下观察到绿色荧光表达;从mRNA和蛋白水平证实,GPC3基因在转染后的MHCC-97L中成功表达;MTT生长曲线证实MHCC97-L/GPC3较其他两株细胞增殖显著加强(P<0.001)。结论成功构建pEGFR-c3-GPC3真核表达载体;筛选出稳定表达GPC3基因的MHCC97-L/GPC3细胞株;GPC3基因表达升高可促进肝癌细胞株MHCC97-L的增殖。     

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…