脑肿瘤干细胞与神经上皮肿瘤病理级别的相关性

目的：探讨脑肿瘤干细胞（brain tumor stem cells，BTSCs）的分离、培养及鉴定的方法，研究BTSCs比例与人脑神经上皮肿瘤病理级别之间的相关性。方法：将人神经上皮肿瘤细胞接种于含生长因子的无血清培养基中，使其中的脑肿瘤干细胞增殖和传代，利用细胞免疫荧光及免疫组化法检测干细胞的特异性标记物Nestin和CD133的表达；采用免疫组化（SABC法）研究46例脑肿瘤及5例正常脑组织CD133的表达，并与肿瘤病理分级进行相关性分析。结果：可以从人神经上皮肿瘤病例中分离培养出BTSCs；BTSCs可以在体外增殖并传代，并且表达干细胞的特异性标记物Nestin和CD133；CD133在正常的脑组织中未见表达，在各级神经上皮肿瘤中均有表达，各级别肿瘤中的阳性细胞比例差异有显著性（P〈0．01），CD133阳性细胞比例与肿瘤的恶性程度呈正相关（P〈0．01）。结论：各种病理类型神经上皮肿瘤组织中均存在干细胞特征的细胞，CD133的表达与肿瘤级别呈正相关。     

神经上皮来源脑肿瘤中肿瘤干细胞的分离培养与鉴定

目的建立人神经上皮来源脑肿瘤中肿瘤干细胞的体外分离、培养体系,观察其分化现象,鉴定其特异性标志物,为脑肿瘤干细胞的进一步研究打下基础。方法将26例脑肿瘤手术中取出的脑肿瘤细胞,以2×10^5／ml的密度接种于含生长因子的无血清培养基培养,使其中的脑肿瘤干细胞增殖,并进行单克隆形成实验及溴脱氧尿苷掺入实验;然后将增殖形成的脑肿瘤干细胞球接种于无生长因子的含血清培养基诱导分化;利用免疫荧光和免疫组化法检测脑肿瘤干细胞在细胞培养和组织切片中CD133和巢蛋白的表达。结果各类肿瘤标本中仅有少数肿瘤细胞能够在无血清培养基中存活并悬浮生长,并增殖形成克隆性脑肿瘤干细胞球,传代后脑肿瘤干细胞保持很强的自我更新和增殖能力;在含血清培养基中脑肿瘤干细胞贴壁分化;体外培养中脑肿瘤干细胞表达神经干细胞的特异性标志物CD133和巢蛋白,在肿瘤组织切片中存在CD133阳性细胞,其阳性表达率为（21．0±6．2）％～（38．0±7．0）％。结论人神经上皮来源脑肿瘤组织中存在少量具有自我更新和增殖能力、表达CD133和巢蛋白的脑肿瘤干细胞,并能在体外将其分离、培养和诱导分化;脑肿瘤干细胞有可能成为脑肿瘤基础研究的新平台和临床治疗的重要靶细胞。     

免疫磁珠法分选人脑胶质瘤干细胞的体外培养及鉴定

目的:旨在从人脑胶质瘤组织和细胞株中分离脑胶质瘤干细胞、进行体外培养并对其干细胞特性加以鉴定,以期对后续个体化敏感药物化疗奠定基础。方法:采用以CD133为标志的免疫磁珠法从人脑胶质瘤组织和细胞株中分离脑胶质瘤干细胞并进行体外培养,通过免疫荧光技术检测干细胞标志物CD133、Nestin,干细胞特性加以鉴定。结果:新鲜人脑胶质母细胞培养得到悬浮生长的肿瘤干细胞球,其能在体外自我更新、增殖,形成新的克隆性细胞球,保持连续稳定的传代,能表达神经干细胞表面标志物CD133。在含血清培养基中能够分化为与原肿瘤组织相似的贴壁生长的细胞。结论:胶质母瘤细胞株中存在的一小部分CD133+胶质瘤细胞具有干细胞的属性,并能稳定传代增生。     

胶质瘤细胞中干细胞分离的实验研究

目的研究应用无血清培养基悬浮法培养U-251胶质瘤细胞系的方法;并分离该细胞系中的脑肿瘤干细胞,鉴定其特异性标志物CD133+的表达并观察其生长和分化特征.方法应用培养大鼠神经干细胞的培养基和培养方法,在无血清培养基中采用悬浮法培养人U-251胶质瘤细胞系;再将分离获得的悬浮生长的脑肿瘤干细胞利用免疫荧光法检测脑肿瘤干细胞特异性标志物CD133+在脑肿瘤干细胞球中的表达.再将脑肿瘤干细胞接种于含血清培养基,观察其分化生长和分化特征.结果 U-251胶质瘤细胞系中有约(2.56±0.2)%的肿瘤细胞在无血清培养基中能够存活,增殖,形成自由漂浮的细胞球;其细胞表达CD133+神经干细胞的标志物,并可连续传代,若重新接种于含血清培养基中可重新贴壁分化,贴壁分化后细胞形态与直接在含血清培养基中培养的U-251胶质瘤细胞系无明显差别.结论用悬浮无血清培养法从U-251人胶质瘤细胞系中成功培养出脑肿瘤干细胞,其肿瘤干细胞能够产生为多种细胞形态的分化细胞,脑肿瘤干细胞的分离培养成功对脑肿瘤的基础和临床研究具有重要价值     

TJ905胶质瘤细胞系中脑肿瘤干细胞的培养、分离与鉴定

目的培养恶性胶质瘤TJ905细胞,并从中分离、培养和鉴定肿瘤干细胞,观察干细胞生长特征,为脑肿瘤干细胞的进一步研究奠定基础。方法将TJ905细胞置于含血清培养基中培养;将TJ905细胞置于含生长因子的无血清培养基中培养,待其形成悬浮生长的细胞球后用免疫磁珠分离获取CD133+细胞。用细胞免疫荧光染色对肿瘤干细胞及其分化细胞进行鉴定。结果在恶性胶质瘤细胞系TJ905中成功分离出肿瘤干细胞,培养于无血清培养液中,细胞呈球形、悬浮生长,具有很强的繁殖与自我更新能力;将该细胞置于含血清的培基中可分化;免疫荧光染色显示脑肿瘤干细胞中神经上皮干细胞蛋白(Nestin)表达阳性。结论培养的恶性胶质瘤细胞系TJ905中有脑肿瘤干细胞的存在,并且能在体外将其培养、分离及诱导分化,可为肿瘤干细胞的研究提供便利条件。     

MRP-1、MDR1、GST-π mRNA在CD133~+肿瘤干细胞中的表达分析

目的分离鉴定CD133~+肿瘤干细胞,初步分析MRP-1、MDR1、GST-π在CD133~+胶质瘤干细胞多药耐药性中的作用。方法采用胶质瘤U+251细胞系、磁珠分离,培养CD133~+胶质瘤干细胞,RT-PCR技术分析MRP-1、MDR1、GST-π在CD133~+胶质瘤干细胞中的表达。结果培养的胶质母细胞瘤干细胞表达干细胞标记物Nestin,分化后细胞表达GFAP、β-tubulin;MRP-1、MDR1、GST-π在CD133~+胶质瘤干细胞中呈高表达,与CD133-胶质瘤干细胞比较差异有统计学意义(P0.05)。结论培养、鉴定胶质母细胞瘤肿瘤干细胞、MRP-1、M DR1、GST-π在CD133~+胶质瘤干细胞中呈高表达,为下一步研究奠定基础。     

胶质瘤干细胞分离鉴定及其化疗敏感性研究

目的:探讨胶质瘤干细胞的分离、培养、鉴定及其与胶质瘤细胞化疗敏感性方面的差异.方法:将3株人胶质瘤细胞SHG-44、U251、U87MG分别接种于无血清培养基,从中分离出各自胶质瘤干细胞;扫描电镜观察胶质瘤干细胞的形态;RT-PCR法检测胶质瘤干细胞CD133、GFAP的表达;CCK8法检测化疗药物顺铂及长春新碱分别作用后胶质瘤细胞及其干细胞活性的变化.结果:从3株胶质瘤细胞中分离出的胶质瘤干细胞,可在体外增殖及传代,表达特异性标志物CD133,不表达GFAP.CCK8法检测出胶质瘤干细胞较胶质瘤细胞更为耐药(均P＜0.05).结论:胶质瘤干细胞较胶质瘤细胞化疗敏感性降低.     

组织块法体外培养人胚胎神经干细胞

目的：探讨体外分离、培养人胚胎脑组织神经干细胞的实用方法。方法：从人胚胎脑额叶皮层分离组织块，采用DMEM／F12＋N2无血清培养基，进行体外培养、传代。采用免疫细胞化学法鉴定神经干细胞和分化后的其他细胞表型。结果：从人胚胎脑额叶皮层分离的细胞具有增殖和多向分化潜能，可进行传代培养，获得的细胞中大部分为Nestin表达阳性的细胞。诱导分化后可以出现NSE、GFAP表达阳性的细胞。结论：用上述方法分离培养的细胞具有神经干细胞的特性，并表达Nestin蛋白，认为是神经干细胞。此方法是一种具有实用价值的体外培养人胚胎脑神经干细胞的技术。     

人HCT116结肠癌细胞系中CD133+结肠癌干细胞分离及鉴定

目的 从人结肠癌细胞系(HCT116)中分离鉴定结肠癌干细胞,并观察其体外生物学特性.方法 利用RT-PCR、免疫荧光、流式细胞等技术检测HCT116细胞中CD133的表达情况;利用免疫磁珠分选技术分选CD133+细胞;采用无血清培养基培养分选后的CD133+细胞鉴定其体外增殖特性;采用含血清培养基诱导干细胞克隆球分化并采用免疫组化检测CK20表达情况鉴定其分化特性.结果 HCT116细胞系中存住CD133+细胞,免疫磁珠能有效的分选CD133+细胞,分选后的阳性含量为91.92%,分选后的阳性细胞具有典型干细胞的体外增殖、分化特性.结论 CD133+结肠癌干细胞在体外具有和普通干细胞类似的无限增殖、自我更新和分化能力,同时它们也表达ABCG2等耐药相火蛋白.     

神经上皮肿瘤中脑肿瘤干细胞的分离培养和原发性多药耐药机制分析

目的 探讨ABC家族转运体耐药基因MDK1、MRP1和DNA修复酶耐药基因MGMT在脑肿瘤干细胞原发性多药耐药中的作用机制.方法 以CD133为脑肿瘤干细胞特异性标志物,利用免疫磁珠法分离30例人脑神经上皮肿瘤标本中的CD133+与CD133-细胞,并进行CD133+细胞的体外扩增培养,传代与鉴定.应用半定量RT-PCK法检测CD133+与CD133-细胞中耐药蛋白MDR1、MRP1、MGMT的活性表达情况.结果 免疫磁珠法能有效的分选出脑肿瘤干细胞,该类细胞具有异常的增殖能力.MDR1、MRP1与MGMT耐药基因在CD133+细胞中高度表达,并与肿瘤的病理级别正相关,其总体水平分别为CD133-细胞的16.1、19.6及34.0倍.并非所有的脑肿瘤干细胞均有耐药基因的表达,MDR1、MRP1与MGMT的总体阳性表达率分别为76.7%、86.7%t和73.3%.ABC家族转运体耐药基因和DNA修复酶耐药基因在CD133+细胞中的表达有明显的相关性.结论 神经上皮肿瘤中存在一定比例的CD133+脑肿瘤干细胞,免疫磁珠法能有效的分选出CD133+细胞.在异质性肿瘤中仅有小部分亚群细胞(CD133+脑肿瘤干细胞)具有原发耐药性.而ABC家族耐药蛋白和DNA修复酶在脑肿瘤干细胞中的共同过度表达是临床上脑肿瘤多药耐药的主要原因之一,脑肿瘤干细胞是神经上皮肿瘤原发性化疗耐药的根源和关键性治疗标靶.     

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…