三氧化二砷对肿瘤血管内皮细胞作用的研究

目的：探讨三氧化二砷（As2O3）对肿瘤血管内皮细胞生长和功能的抑制作用及其相关的抗血管形成机制。方法：采用肝癌HepG2细胞上清诱导人脐静脉内皮细胞（human umbilical vein endothelial cells,HUVEC）成为肿瘤血管内皮细胞（tumor-derived endothelial cells,Td-EC）,通过细胞迁徙实验流式细胞术、流式细胞术、血管形成实验检测As2O3对Td-EC细胞生物学特征的影响。结果：As2O3在体外对Td-EC有显著促进凋亡的作用,并能抑制其迁移和血管的形成,较在同样条件下对HUVEC的凋亡、迁移和血管形成作用显著。结论：As2O3在体外可特异地抑制Td-EC生长及功能。     

脑胶质瘤细胞外泌体对血管新生的影响及其机制

目的 探讨脑胶质瘤细胞外泌体对血管新生的影响及其机制。方法 差速离心法提取人脑胶质瘤细胞U373和U87以及脑胶质细胞HEB所分泌的外泌体,透射电镜观察外泌体结构,Western blot鉴定外泌体标志蛋白;将0.2 μg/μL的外泌体和等体积PBS分别与人脐静脉内皮细胞HUVECs共孵育,CCK-8法检测细胞增殖能力,Transwell实验检测细胞迁移能力,小管形成实验检测体外血管生成,流式细胞术检测细胞凋亡水平,Western blot检测凋亡蛋白Bax、Caspase3、Caspase9、VEGF、VEGFR2的表达及AKT的磷酸化水平。结果 提取的外泌体符合其特征;相比于PBS组,与HEB-exo共孵育后,HUVECs增殖、迁移、小管形成、细胞凋亡均无显著变化（P>0.05）;而与U373-exo和U87-exo共孵育后HUVECs增殖、迁移、小管形成能力均显著提高（P<0.05）;细胞凋亡水平显著降低（P<0.001）,Bax、Caspase3、Caspase9表达显著下降（P<0.001）,VEGF、VEGFR2的表达以及AKT的磷酸化水平显著提高（P<0.001）。结论 脑胶质瘤细胞外泌体能够激活调控VEGF/AKT信号通路而促进血管新生。     

茶氨酸对内皮细胞生长及肺癌血管生成的影响

目的探讨茶氨酸对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)增殖、凋亡、小管形成及肺腺癌移植瘤血管生成的影响。方法 HUVECs分为6组,予以6种不同浓度茶氨酸处理,MTT法检测细胞生长抑制率,流式细胞仪检测细胞凋亡率,应用三维培养反映体外小管形成能力;建立异位肺腺癌裸鼠模型,分为茶氨酸组和对照组,每组6只,茶氨酸组予以茶氨酸腹腔注射,对照组予以等体积生理盐水腹腔注射。检测肿瘤中VEGF的表达及肿瘤微血管密度(microvessel density,MVD),明确茶氨酸对肿瘤血管生成的影响。结果随着茶氨酸浓度的增加,HUVECs的生长抑制率及凋亡率增加(P<0.05,P<0.01);茶氨酸浓度依赖性地抑制体外小管形成(P<0.05,P<0.01);茶氨酸组较对照组肿瘤中VEGF表达、MVD明显下降(P<0.05)。结论茶氨酸呈浓度依赖性地抑制内皮细胞增殖,诱导内皮细胞凋亡,抑制内皮细胞体外小管形成。茶氨酸能抑制裸鼠移植瘤新生血管形成。     

三七皂苷R1对氧化型低密度脂蛋白诱导血管内皮细胞凋亡的保护作用

目的探讨三七皂苷R1（NGR1）对氧化型低密度脂蛋白（ox-LDL）诱导血管内皮细胞凋亡的保护作用及其机制。方法将体外培养人脐静脉内皮细胞（HUVECs）分为对照组、损伤组（ox-LDL组）和4种浓度NGR1治疗组（ox-LDL+不同浓度NGR1组）;采用CCK-8法检测NGR1的药物毒性以及各组HUVECs的增殖活性,采用Westernblot法检测Bcl-2、Bax及Cleaved-caspase-3凋亡相关蛋白表达情况,免疫荧光染色检测Cleaved-caspase-3凋亡情况,钙黄绿素细胞膜标记染色检测细胞黏附能力,Transwell迁移试验检测细胞迁移能力。结果与对照组比较,损伤组HUVECs的凋亡率和Cleaved-caspase-3表达水平显著升高,并且HUVECs的增殖活性、细胞黏附能力和迁移能力均显著降低（均P＜0.05）。与损伤组比较,NGR1治疗组HUVECs的凋亡率和Cleaved-caspase-3表达水平均随着NGR1浓度的增加而逐渐下降,HUVECs的增殖活性、细胞黏附能力和迁移能力均随着NGR1浓度的增加而逐渐升高（均P＜0.05）。结论NGR1通过抑制ox-LDL诱导的HUVECs细胞增殖活性降低、Cleaved-caspase-3表达上调而引起的细胞凋亡增多、细胞黏附和迁移数量减少,从而发挥其对ox-LDL损伤的血管内皮细胞的保护作用。     

FOXC2调控DLL4/Notch1信号通路对乳腺癌MCF-7细胞血管生成的影响

目的：探讨转录因子FOXC2对乳腺癌MCF-7细胞血管生成的影响,阐明FOXC2促进肿瘤血管生成的作用机制。方法：应用FOXC2慢病毒基因转染技术将FOXC2基因和空载体基因分别转染至乳腺癌MCF-7细胞株中,获得稳定转染细胞株;实验分为未转染组、空载体组和过表达组。采用Matrigel基质胶血管形成实验和Transwell小室实验检测各组细胞上清作用下人脐静脉内皮细胞（HUVECs）成管能力和迁移能力,RT-PCR和Western blotting法检测各组细胞中FOXC2、DLL4和Notch1 mRNA和蛋白表达。结果：与未转染组和空载体组比较,过表达组MCF-7细胞上清诱导HUVECs闭合小管数和迁移细胞数增加（P<0.05）;FOXC2、DLL4和Notch1 mRNA和蛋白相对表达水平明显增加（P<0.05）。结论：MCF-7细胞中过表达FOXC2能显著增加HUVECs的成管能力和迁移能力,其机制可能是通过Notch信号通路来实现的。     

干扰人乳腺癌MDA-MB-231细胞 EPCR表达对内皮细胞增殖､迁移及脉管形成的影响

目的 :通过观察干扰乳腺癌MDA-MB-231细胞内皮蛋白C受体(endothelial protein C receptor,EPCR)表达对内皮细胞的增殖、迁移、脉管形成能力的变化,分析EPCR在肿瘤血管形成中的作用。方法:采用si RNA方法降低人乳腺癌MDA-MB-231细胞EPCR的表达,通过RT-PCR及Western blot技术检测EPCR干扰效果,实验分为EPCR干扰组、无关序列组及未处理组。制备肿瘤条件培养基模拟肿瘤微环境培养HUVECs细胞,通过CCK-8法检测各组内皮细胞增殖能力,Transwell小室检测内皮细胞迁移能力,Matrigel检测内皮细胞脉管形成能力。结果:与未处理组及无关列序组相比,EPCR干扰组细胞的增殖、迁移及脉管形成能力均明显降低(P<0.05)。结论:干扰乳腺癌MDA-MB-231细胞EPCR的表达能够抑制内皮细胞增殖、迁移及脉管形成能力,提示EPCR可能在肿瘤血管形成中起重要作用。     

松香酸通过上调miR-30a-3p表达增强缺氧诱导的HUVECs血管生成

目的探讨松香酸对急性心肌梗死(AMI)后内皮细胞血管生成及迁移的作用与机制。方法在体外构建人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HUVECs)细胞缺氧模型;通过Matrigel小管形成实验检测HUVECs的血管生成情况;Transwell及伤口愈合实验检测HUVECs的细胞迁移;Western blot检测血管内皮生成因子A(VEGFA)的表达情况;qRT-PCR检测miR-30a-3p的转录水平。结果成功构建细胞缺氧模型;Matrigel小管形成实验显示缺氧抑制HUVECs的血管生成及细胞迁移,而利用松香酸处理缺氧细胞后,可以促进其血管生成和细胞迁移;Western blot结果显示,松香酸能够促进VEGFA的表达;进一步利用qRT-PCR检测松香酸处理后细胞内miR-30a-3p水平,发现松香酸可明显提高miR-30a-3p水平,而在细胞内转染miR-30a-3p inhibitor后,松香酸促进HUVECs的血管生成和细胞迁移作用被抑制。结论松香酸可能通过调节miR-30a-3p的表达,增强HUVECs的血管生成与细胞迁移。     

miRNA-195抑制剂对高糖培养血管内皮细胞生物学行为的影响及机制

目的：探讨高糖环境下miRNA-195 抑制剂对血管内皮细胞行为的影响及机制。方法：培养人脐静脉血管内皮细胞（HUVECs）ECV304,分别用高糖（HG）、miRNA-195模拟质粒（mimic）、miRNA-195抑制质粒（inhibitor）、Sirt1激动剂白藜芦醇（RES）处理细胞株,CCK-8试剂盒检测增殖活力,RT-PCR检测miRNA-195及Sirt1 mRNA水平,Western blot检测Sirt1、p-FoxO1、FoxO1、Caveolin-1、血管内皮生长因子（VEGF）蛋白表达水平,划痕实验检测HUVECs的迁移能力,transwell小室实验检测细胞侵袭能力,Matrigel胶检测HUVECs的成管能力。结果：HG促进HUVECs的增殖、迁移、侵袭,增强成管能力（P ＜0.05）;而miRNA-195inhibitor则明显抑制HUVECs的增殖、迁移,降低成管能力（P ＜0.05）。结论：miRNA-195 inhibitor或许通过miRNA-195/Sirt1/FoxO1/Caveolin-1/VEGF通路影响HG环境下HUVECs的生物学功能。     

芍药醇对叔丁基过氧化氢损伤的血管内皮细胞的保护作用

目的：探讨芍药醇（PAE）对叔丁基过氧化氢（TBHP）损伤的血管内皮细胞的保护作用及其机制。方法：体外培养人脐静脉内皮细胞（HUVECs）分为对照组（Control组）、造模组（TBHP组）和PAE处理组（各浓度PAE+THBP组）;用CCK8法及乳酸脱氢酶（LDH）检测各组HUVECs细胞增殖抑制情况,Transwell实验检测细胞的迁移能力,流式细胞仪和Cleaved-caspase 3免疫荧光染色检测HUVECs细胞凋亡情况。结果：与Control组比较,TBHP组细胞存活率和细胞迁移数明显降低,细胞凋亡率、LDH释放量和Cleaved-caspase 3蛋白的表达明显升高（均P＜0.05）。与TBHP组比较,PAE处理组细胞的存活率和细胞迁移数均随着给药浓度的增加而逐渐升高,细胞凋亡率、LDH释放量和Cleaved-caspase 3蛋白的表达均随着给药浓度的增加而逐渐下降（均P＜0.05）。结论：PAE通过抑制TBHP诱导的HUVECs细胞存活率降低、LDH释放量升高、细胞凋亡增多、Cleaved-caspase 3蛋白表达的上调,发挥其对TBHP损伤的血管内皮细胞的保护作用,且呈剂量依赖性。     

三氧化二砷对血管内皮细胞增殖和周期的影响

目的:观察As2O3对血管内皮细胞增殖、凋亡和细胞周期的干扰,探讨As2O3对血管内皮细胞生长的直接影响。方法:血管内皮细胞EVC-304体外培养,As2O3干预。采用MTT测定细胞活性,流式细胞仪测定细胞周期、细胞凋亡。结果:ECV-304细胞经过As2O3干预,其存活率明显降低,并且呈剂量依赖性。As2O3干预后,进入S期的细胞减少,细胞周期被阻在G1期,细胞可能在G1期进入凋亡。As2O3诱导早期凋亡是对照组的2.88～5.1倍,并且呈剂量依赖性;其晚期凋亡是对照组的1.17～1.67倍。结论:As2O3可直接干扰血管内皮细胞的细胞周期,阻滞细胞在G1期,并诱导细胞凋亡,进而抑制血管内皮细胞的增殖。     

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…