缺氧对小鼠小胶质细胞IRAK-1表达的影响

目的观察缺氧后小鼠小胶质细胞株N9对白细胞介素-1受体相关激酶-1(interleukin-1 receptor associated kinase-1,IRAK-1)的mRNA、蛋白表达的变化,探讨中枢神经系统炎症反应的相关机制。方法将培养的N9细胞置于缺氧(3%O2)条件下培养3、8、24h,用RT-PCR及Western印迹方法检测IRAK-1的mRNA和蛋白表达的变化,用酶联免疫吸附法检测培养上清液肿瘤坏死因子α(TNF-α)含量。结果N9细胞IRAK-1的mRNA和蛋白的表达随缺氧时间延长而逐渐增高,3h开始增加(P<0.01),8h达高峰(P<0.01),24h下降接近正常水平(P>0.05)。培养上清液TNF-α的含量也有相似的变化趋势。结论缺氧可以影响IRAK-1的mRNA和蛋白表达,提示中枢神经系统内存在对免疫反应起着调控作用的TLR/IL-1R家族信号传导系统,并且调控下游炎症反应。     

缺氧条件下大鼠神经元白细胞介素-1受体相关激酶-1的表达变化

目的观察大鼠神经元B35细胞株中白细胞介素-1受体相关激酶-1(interleukin-1 receptor-associated kina-ses-1,IRAK-1)mRNA及蛋白在缺氧后表达量的变化,并探讨其变化与神经元中TNF-α表达的关系。方法将培养的B35细胞置于低氧(3%O2、5%CO2、92%N2)条件下培养1、3、6、12、24、48、72 h,应用RT-PCR方法检测IRAK-1 mRNA表达的变化;Western blot检测IRAK-1蛋白含量的变化;激光扫描共聚焦观察IRAK-1在细胞内的分布及含量变化;酶联免疫吸附法检测培养液上清肿瘤坏死因子-α(TNF-α)的含量,并与IRAK-1蛋白进行相关分析。结果B35细胞IRAK-1 mRNA表达与正常对照相比,1 h开始增加[(0.786±0.042),P<0.05],3 h增加明显[(0.947±0.055),P<0.05],6 h达到高峰(1.245±0.077),P<0.05],12 h仍维持在较高水平[(0.911±0.041),P<0.05],24 h降至正常水平以下但无明显差异[(0.596±0.047),P>0.05],48...     

高浓度氧暴露对N9小胶质细胞功能的影响

目的探讨常压高浓度氧暴露对N9小胶质细胞功能的影响。方法体外培养N9小胶质细胞,分别在900 ml/L高浓度氧中暴露2、6、12、16、24、48 h后(1)流式细胞术检测细胞存活率及凋亡率;(2)DCFH-DA荧光探针检测活性氧(ROS)含量的变化;(3)RT-PCR检测Toll样受体4(TLR4)mRNA表达变化;(4)ELISA检测N9细胞培养上清中IL-1β和TNF-α的含量;(5)免疫荧光化学染色检测TLR4蛋白的表达变化。结果流式细胞术结果示,细胞凋亡率在高氧暴露12 h后明显高于空气对照组(P<0.05),16 h达高峰(P<0.01);高氧暴露2 h后,细胞内ROS含量明显增加,呈一定时间依赖性;TLR4 mRNA及蛋白表达水平明显高于空气对照组(P<0.05);高氧暴露后细胞上清中TNF-α及IL-1β含量随暴露时间延长而增加,均在6 h开始升高,12~16 h达高峰(P<0.05),IL-1β增高较TNF-α更为明显。结论高氧暴露后N9小胶质细胞TLR4通路激活,产生大量神经毒性因子(ROS、IL-1β及TNF-α),细胞凋亡增加。     

高浓度氧暴露对N9小胶质细胞功能的影响

摘要：目的探讨常压高浓度氧暴露对N9小胶质细胞功能的影响。方法体外培养N9小胶质细胞,分别在900 ml/L高浓
度氧中暴露2、6、12、16、24、48 h后（1）流式细胞术检测细胞存活率及凋亡率;（2）DCFH-DA荧光探针检测活性氧（ROS）含
量的变化;（3）RT-PCR检测Toll样受体4（TLR4）mRNA表达变化;（4）ELISA检测N9细胞培养上清中IL-1β和TNF-α的含
量;（5）免疫荧光化学染色检测TLR4蛋白的表达变化。结果流式细胞术结果示,细胞凋亡率在高氧暴露12 h后明显高于
空气对照组（P<0.05）,16 h达高峰（P<0.01）;高氧暴露2 h后,细胞内ROS含量明显增加,呈一定时间依赖性;TLR4 mRNA
及蛋白表达水平明显高于空气对照组（P<0.05）;高氧暴露后细胞上清中TNF-α及IL-1β含量随暴露时间延长而增加,均在
6 h开始升高,12~16 h达高峰（P<0.05）,IL-1β增高较TNF-α更为明显。结论高氧暴露后N9小胶质细胞TLR4通路激活,
产生大量神经毒性因子（ROS、IL-1β及TNF-α）,细胞凋亡增加。
     

缺氧条件下大鼠单核细胞NF-κB活性及TNF-α、IL-1β表达变化规律的研究

目的研究缺氧培养的大鼠外周血单核细胞核转录因子κB(NF-κB)结合活性及肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)表达的变化规律,探讨缺氧与全身炎症反应综合征的关系,为进一步研究老年多器官功能障碍综合征的肺启动机制奠定基础.方法采用明胶法分离大鼠外周血单核细胞,于低氧条件下(3%O2, 5%CO2, 92%N2)培养0、1、3、6、9、12、24 h后,收集细胞及培养液上清,分别采用电泳迁移分析法(EMSA)、逆转录PCR(RT-PCR)和酶联免疫吸附试验(ELISA)检测其NF-κB活性及TNF-α、IL-1β表达量.结果缺氧1～12 h,NF-κB结合活性及TNF-α、IL-1β表达量均显著高于正常(P<0.01),其峰值见于缺氧6 h.缺氧24 h,上述指标与正常无显著差异(P＞0.05).NF-κB结合活性与TNF-α、IL-1β mRNA和蛋白表达水平显著相关(P<0.01,P<0.05).结论缺氧可显著增强大鼠外周血单核细胞的NF-κB结合活性,并可诱导其产生大量的促炎症因子TNF-α、IL-1β,这些变化可能与急性呼吸窘迫综合征时血浆中大量促炎症因子持续存在密切相关.     

体外培养大鼠视网膜神经节细胞缺氧条件下HIF-1α动态表达及意义

目的探讨体外培养Long Evans大鼠视网膜神经节细胞缺氧条件下缺氧诱导因子1α（hypoxia induction factor 1α，HIF-1α）表达变化及损伤机制。方法正常对照组视网膜神经节细胞在常规条件下、缺氧组在1％氧浓度下分别培养1、3、12、24h。采用RT—PCR法检测神经节细胞HIF-1α mRNA水平，蛋白质免疫印迹法测定HIF-1α蛋白水平表达，免疫荧光法进行HIF-1α细胞内定位检测，分别采用黄嘌呤氧化酶法、硫代戊巴比妥酸法、酶标双抗夹心免疫吸附法ELISA方法测定培养液SOD活性、MDA含量及TNFoa水平检测。结果正常对照组神经节细胞未见HIF-1α mRNA及蛋白表达，缺氧组HIF-1α mRNA水平、蛋白表达随缺氧时间延长显著升高（P〈0．01），其峰值分别在缺氧后3h和12h。免疫荧光法见缺氧组HIF-1α表达于神经节细胞胞浆。缺氧组细胞培养液SPD活性随缺氧时间时间延长显著降低，MDA含量及TNF-α含量显著升高，差异有统计学意义（P〈0．05、P〈0．01）。结论体外培养视网膜神经节细胞在缺氧条件下其HIF-1α mRNA水平及蛋白表达呈上升后降低趋势，其表达变化与缺氧时培养液中脂质过氧化反应逐渐增强、抗氧化能力降低及炎性因子水平上升有关。     

神经肽Y对小神经胶质细胞活化状态和生成TNF-α的影响及其作用机制

目的：探讨神经肽 Y（neuropeptide Y,NPY）对原代小神经胶质细胞生物活性及生成肿瘤坏死因子α（tumor necrosis factor-α,TNF-α）的影响及其作用机制。方法培养原代大鼠皮层小胶质细胞,将细胞分为 Control组、脂多糖（lipopolysaccharide,LPS）组、NPY＋LPS 组、NPY 组和 BIBP3226＋NPY＋LPS 组,分别用无血清胶质细胞培养液、含 LPS 的无血清胶质细胞培养液、含 NPY 和 LPS 的无血清胶质细胞培养液、含 NPY 的无血清胶质细胞培养液和含 BIBP3226、NPY 和 LPS 无血清胶质细胞培养孵育细胞6 h。经免疫细胞化学荧光染色后,显微镜下观察小胶质细胞的形态学变化。ELISA 方法检测培养液中 TNF-α蛋白含量,RT-PCR 方法检测小胶质细胞中TNF-αmRNA 表达水平。结果 LPS 组 TNF-α的含量及细胞中 TNF-αmRNA 表达水平显著高于 Control 组（P ＜0．05）,小胶质细胞处于活化状态;LPS＋ NPY 组与 LPS 组相比,TNF-α蛋白和 mRNA 表达水平明显降低（P ＜0．05）,小胶质细胞活化水平降低;BIBP3226＋NPY＋LPS 组与 LPS＋NPY 组相比,TNF-α蛋白和 mRNA 表达水平显著增高（P ＜0．05）;LPS 组与 BIBP3226＋NPY＋LPS 组差异无统计学意义;NPY 组与 Control 组差异无统计学意义。结论 NPY 可以降低小神经胶质细胞的生物活性,抑制其生成 TNF-α,作用机制可能与 NPY Y1受体有关。     

Exendin-4对肿瘤坏死因子-α诱导的单核细胞趋化蛋白-1表达的影响

目的观察exendin-4对大鼠肾小球系膜细胞单核细胞趋化蛋白-1(MCP-1)、纤维连接蛋白(FN)表达的影响。方法试验分组:对照组、肿瘤坏死因子-α(TNF-α)组(10 ng/ml)、TNF-α(10 ng/ml)+E1组(1 nmol/L exendin-4)、TNF-α(10 ng/ml)+E5组(5 nmol/L exendin-4)、TNF-α(10 ng/ml)+E10组(10 nmol/L exendin-4)。收集24 h细胞,提取RNA,采用实时荧光定量检测细胞内MCP-1 mRNA的表达;分别在24、48 h收集各组细胞培养上清液,采用ELISA检测培养细胞上清中MCP-1、FN的浓度。结果孵育24 h时,TNF-α组MCP-1 mRNA和细胞培养上清MCP-1、FN表达较对照组明显增加(P<0.01);与TNF-α组比较,TNF-α+E5组、TNF-α+E10组细胞内MCP-1 mRNA和培养上清内MCP-1、FN含量均明显降低;孵育48 h时,TNF-α组细胞培养上清MCP-1、FN表达较对照组明显增加(P<0.01);与TNF-α组比较,TNF-α+E1组、TNF-α+E5组、TNF-α+E10组细胞培养上清内MCP-1、FN含量均明显降低;并且随着时间的延长和exendin-4浓度的增加,培养上清内MCP-1、FN含量降低更加明显。结论 Exendin-4可明显抑制TNF-α诱导的系膜细胞MCP-1 mRNA的表达,并以剂量和时间依赖的方式抑制TNF-α诱导的系膜细胞培养上清内MCP-1、FN的含量,提示exendin-4可能会对肾脏具有一定保护作用。     

IL-10对TNF-α诱导的泡沫细胞三磷酸腺苷结合盒转运体A1表达的抑制作用

目的:观察白介素-10(IL-10)对肿瘤坏死因子-α(TNF-α)诱导的THP-1巨噬细胞源性泡沫细胞三磷酸腺苷结合盒转运体A1(ABCA1)表达的干预作用.方法:THP-1单核细胞经诱导转化为巨噬细胞源性泡沫细胞后随机分为4组:对照组,培养液中不加任何其他物质;TNF-α组,培养液中加10μg/L TNF-α;IL-10组,培养液中加10μg/L IL-10;IL-10 TNF-α组,培养液中加10μg/L IL-10预先孵育2 h后,再加10μg/L TNF-α.采用RT-PCR和Western-Blot检测各组0 h、6 h、12 h、24 h、48 h ABCA1 mRNA和蛋白表达.结果:TNF-α组ABCA1 mRNA和蛋白表达呈时间依赖性降低(F=782.94,F=768.12,P均＜0.05).IL-10组ABCA1 mRNA表达在24 h和48 h有轻微增加(P＜0.05),但ABCA1蛋白表达在各时间点无明显变化.IL-10 TNF-α组ABCA1 mRNA和蛋白表达也呈时间依赖性降低(F=697.23,F=749.64,P均＜0.05),但同TNF-α组相比,其下降程度有所减轻(P＜0.05).结论:IL-10可部分抑制TNF-α所引起THP-1巨噬细胞源性泡沫细胞ABCA1表达的下调.     

牛磺酸预处理抑制IRAK-4信号通路对大鼠移植肝脏再灌注损伤的保护机制

目的 观察牛磺酸预处理后SD大鼠移植肝脏的白细胞介素-1受体相关激酶-4(interleukin-1 receptor aasociated kinase-4,IRAK-4)表达水平的变化,探讨牛磺酸预处理抑制肝脏缺血再灌注损害的相关机制.方法 雄性SD大鼠128只,完全随机化分为生理盐水对照组(NS组)和牛磺酸预处理组(TA组),每组64只,用Kamada's袖套法经行肝移植.NS组切取供肝前10 min经腰静脉注射生理盐水1.5 ml,TA组切取供肝前10 min经腰静脉注射牛磺酸(300 mmol/L)1.5 ml.于再灌注后0、60 min及180 min,蛋白免疫印记法及实时荧光定量PCR测定肝组织中IRAK-4 mRNA和蛋白表达水平;凝胶电泳迁移率分析法(EMSA)检测肝组织NF-κB活性及血清TNF-α含量;全自动血清自动生物化学仪检测血清转氨酶;光镜切片检测肝脏组织学.结果 牛磺酸预处理能明显提高大鼠1周生存率(P＜0.01),改善肝功能,减轻肝组织病理学改变.再灌注后0 min,血清转氨酶、IRAK-4 mRNA与蛋白表达水平、NF-κB活性以及TNF-α含量2组间比较差异无统计学意义(P＞0.05);NS组各项指标水平于再灌注60 min出现峰值,但TA组各项指标水平明显低于NS组(P＜0.01);再灌注后180 min,2组IRAK-4 mRNA与蛋白表达水平、NF-κB活性以及TNF-α含量均较60 min时有所下降,且TA组下降水平较NS组更为明显(P＜0.01).结论 牛磺酸对大鼠移植肝脏的缺血再灌注损伤有明显抑制作用,抑制IRAK-4及其下游的NF-κB、TNF-α表达是牛磺酸预处理减轻肝脏缺血再灌注损害的重要机制之一.     

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…