芍药苷减弱真菌葡聚糖诱导的支气管上皮细胞内NLRP3炎性小体活化

目的：探讨脂多糖（lipopolysaccharide,LPS）联合真菌葡聚糖能否活化支气管上皮细胞16HBE内NOD样受体热蛋白结构域相关蛋白3（Nod-like receptor pyrin domain-containing protein 3,NLRP3）炎性小体,芍药苷（paeoniflorin,PF）对该NLRP3炎性小体的活化是否有抑制作用及相关机制。方法：LPS联合真菌葡聚糖建立感染模型,RT-PCR检测细胞内NLRP3、caspase-1、白细胞介素（interleukin,IL）-1β mRNA的表达;ELISA检测细胞上清中IL-1β含量;caspase-1活性检测试剂盒检测胞内caspase-1活化程度;流式检测胞内活性氧（reactive oxygen species,ROS）变化;Western blot检测胞内NLRP3、caspase-1、IL-1β蛋白表达变化。结果：LPS联合真菌葡聚糖联合作用于支气管上皮细胞,胞内NLRP3、caspase-1、IL-1β表达转录加强,细胞上清中IL-1β含量增加,caspase-1活性上调,细胞内ROS升高;PF预作用细胞后,胞内ROS随着药物浓度增加而逐渐降低,NLRP3、caspase-1、IL-1β的转录表达也随之受抑制而下调。结论：LPS联合真菌葡聚糖可有效活化支气管上皮细胞内NLRP3炎性小体,而PF能有效抑制胞内ROS产生从而抑制炎性小体活化、抑制真菌葡聚糖所致的炎性反应。     

NLRP3炎性小体在脂多糖诱导的高糖心肌细胞缺氧复氧损伤中的作用及机制

目的 探究NLRP3炎性小体在脂多糖(lipopolysaccharide, LPS)诱导的心肌细胞高糖缺氧/复氧(hypoxia/reoxygenation, H/R)损伤中的作用及机制。方法 取正常对数期生长的H9C2心肌细胞,随机分为4组,即高糖(H)组(n=3)、高糖+缺氧/复氧(H+H/R)组(n=3)、高糖+LPS+H/R(H+LPS+H/R)组(n=3)和BAY11-7082干预高糖+LPS+H/R(H+LPS+BAY+H/R)组(n=3)。采用CCK-8法检测细胞活力,ROS检测试剂盒测定线粒体氧化应激水平,RT-PCR检测细胞NLRP3、ASC、caspase-1和IL-1β的mRNA表达水平,Western blot法检测细胞NLRP3、ASC、caspase-1和IL-1β的蛋白表达水平。结果 与高糖组比较,H+H/R组细胞活性降低、LDH水平升高、线粒体ROS产生增加,NLRP3、ASC、caspase-1和IL-1β的mRNA表达上调,NF-κB、NLRP3、ASC、caspase-1和IL-1β蛋白水平增高(P<0.05);LPS处理后,H/R的高糖心肌细胞损伤进一步加重(P<0.05),而加用BAY11-7082后,LPS加重的损伤得以改善(P<0.05)。结论 BAY11-7082可以抑制NLRP3炎性小体激活,从而减轻LPS介导的H9C2细胞高糖H/R损伤。     

LPS激活巨噬细胞NLRP3炎性小体的作用研究

目的 探讨脂多糖(LPS)激活巨噬细胞NLRP3炎性小体的作用.方法 用佛波酯诱导人单核细胞(THP-1)分化为巨噬细胞.实验分为对照组、LPS组、LPS+NLRP3 siRNA组、LPS+Scrambled siRNA组.采用免疫荧光染色检测核苷酸结合寡聚化结构域样受体家族pyrin结构域蛋白3(NLRP3)和凋亡相关斑点样蛋白ASC的共定位、Western blotting检测NLRP3蛋白的表达、ELISA检测细胞培养上清白介素1β(IL-1β)的浓度.结果 与对照组相比,NLRP3和ASC蛋白的共定位明显增多,LPS组NLRP3蛋白的表达增强.LPS还能使细胞培养上清IL-1β的浓度显著升高,NLRP3 siRNA干扰后细胞培养上清IL-1β的浓度显著降低.结论 LPS可以促进炎性小体成分NLRP3、ASC的表达,激活NLRP3炎性小体,刺激巨噬细胞分泌炎症因子IL-1β.     

黄芩抑制脂多糖/三磷酸腺苷诱导的内皮细胞NIMA相关蛋白激酶7(NEK7)表达和Nod样受体蛋白3(NLRP3)炎症小体激活

【目的】观察黄芩和黄芩苷对脂多糖(LPS)/三磷酸腺苷(ATP)诱导的人脐静脉内皮细胞(HUVECs)NIMA相关蛋白激酶7(NEK7)和Nod样受体蛋白3(NLRP3)小体表达的影响。【方法】将HUVECs随机分为空白对照组、模型组、阿托伐他汀组、黄芩苷组和黄芩血清组,按分组相应药物预处理细胞24 h,然后用1μg/mL LPS刺激细胞24 h,再用5.5 mmol/L ATP刺激细胞4 h后检测各指标。四甲基偶氮唑盐(MTT)法检测细胞活力,均相竞争法测定细胞上清液中内皮素1(ET-1)的含量,硝酸还原酶法测定细胞上清液中总硝酸盐(NOx)水平,蛋白免疫印迹法检测细胞内NEK7、NLRP3、凋亡相关斑点样蛋白(ASC)、裂解型胱天蛋白酶1(cleaved-Caspase-1)、白细胞介素1β(IL-1β)蛋白的表达。【结果】与模型组比较,黄芩血清组、黄芩苷组、阿托伐他汀组LPS/ATP诱导的细胞活力升高,ET-1水平降低、NOx水平升高,NEK7、NLRP3、ASC、cleaved-Caspase-1、IL-1β的蛋白表达水平降低(P<0.05或P<0.01),且黄芩血清组、黄芩苷组的作用效果优于阿托伐他汀组。【结论】黄芩和黄芩苷可以抑制LPS/ATP诱导HUVECs的NLRP3炎症小体激活引起的细胞焦亡而保护内皮,其机制可能与调控NEK7表达有关。     

降钙素相关基因肽在小鼠巨噬细胞炎症调控中的作用研究

目的 探讨降钙素相关基因肽（CGRP）在炎症调控中的作用,验证CGRP可通过活性氧簇-核苷酸结合低聚体结构域样受体3（ROS-NLRP3）信号通路抑制小鼠巨噬细胞炎症因子的分泌。方法 实验分为对照组、脂多糖（LPS）100?ng/ml组（LPS组）、CGRP 10?ng/ml组（CGRP 10组）、CGPR 30?ng/ml组（CGRP 30组）、CGRP 100?ng/ml组（CGRP 100组）及CGRP 30?ng/ml+LPS 100?ng/ml组（CGRP 30+LPS组）。作用12?h后,分别收集各组细胞的培养上清,酶联免疫吸附试验（ELISA）检测上清液中白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）的蛋白表达。同时收集各组细胞,实时荧光定量聚合酶链反应（qRT-PCR）检测炎症因子IL-1β、TNF-α和炎症复合体NLRP3 mRNA的表达水平。流式细胞术检测各组细胞内活性氧簇（ROS）水平,Western blotting检测ROS-NLRP3信号通路相关蛋白NLRP3、Caspase-1、IL-1β及TNF-α的蛋白表达。结果 与对照组比较,LPS组细胞中IL-1β、TNF-α及NLRP3 mRNA表达水平升高,培养上清液中IL-1β和TNF-α的蛋白表达水平升高,细胞内ROS水平升高,且细胞内NLRP3、Caspase-1、IL-1β及TNF-α的蛋白表达水平升高（P?<0.05）;与LPS组比较,CGRP 10组、CGRP 30组、CGRP 100组、CGRP 30+LPS组细胞中IL-1β、TNF-α及NLRP3 mRNA表达水平降低,培养上清液中IL-1β和TNF-α蛋白表达水平降低,细胞内ROS水平降低,且细胞内NLRP3、caspase-1、IL-1β及TNF-α蛋白表达水平也降低,所有指标中CGRP 30组降低最明显（P?<0.05）。结论 CGRP可降低巨噬细胞内ROS和NLRP3的表达,使炎症因子IL-1β和TNF-α的释放减少,CGRP可能在减弱局部炎症反应中发挥重要的调控作用。     

苦参碱联合Akt信号通路抑制剂影响肾癌细胞的活力与凋亡

目的本研究探讨苦参碱联合蛋白激酶B(Akt)信号通路抑制剂对肾癌细胞增殖活力及凋亡的影响及机制。方法以肾癌细胞为研究对象,用不同浓度的苦参碱作用后,噻唑蓝(MTT)检测细胞增殖活力,计算半数抑制浓度。用半数抑制浓度的苦参碱和Akt信号通路抑制剂分别处理细胞记为苦参碱组和抑制剂组,同时以半数抑制浓度的苦参碱和Akt信号通路抑制剂共同处理后的细胞为联合组,以正常培养的细胞为对照组,MTT检测细胞增殖活力,流式细胞术检测细胞凋亡,激光扫描共聚焦显微镜检测细胞中活性氧(reactive oxygen species, ROS)水平,蛋白质印迹法检测细胞中p38、磷酸化的p38(p-p38)、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(cleaved cysteinyl aspartate specific proteinase 3, cleaved caspase-3)表达。结果苦参碱能够呈浓度依赖的抑制肾癌细胞增殖活力。与对照组比较,苦参碱组、抑制剂组、联合组细胞存活率降低,凋亡率升高,ROS水平升高,p-p38水平、cleaved caspase-3蛋白水平升高。与苦参碱组、抑制剂组比较,联合组胞存活率降低,凋亡率升高,ROS水平升高,p-p38水平、cleaved caspase-3蛋白水平升高。结论苦参碱联合Akt信号通路抑制剂能够抑制肾癌细胞增殖活力,促进肾癌细胞凋亡,ROS和p38信号通路可能是其作用机制之一。     

原花青素对LPS/ATP诱导的NLRP3炎症小体激活及NF⁃κBp65磷酸化的抑制作用

目的：研究原花青素对脂多糖（lipopolysaccharide,LPS）与腺嘌呤核苷三磷酸（adenosine triphosphate,ATP）联合诱导的小鼠小胶质细胞BV2 NOD样受体蛋白3（nucleotide?binding oligomerization domain?like receptor protein 3,NLRP3）炎症小体激活及核因子（nuclear factor,NF）?κBp65磷酸化的抑制作用。方法：以1.0 μg/mL LPS、2.5 μmol/L ATP诱导BV2细胞炎症损伤模型,用不同浓度原花青素（0.1、1.0、2.5、5.0 μg/mL）干预细胞。采用噻唑蓝（thiazolyl blue tetrazolium bromide,MTT）法检测细胞存活率;比色法检测一氧化氮（nitric oxide,NO）释放水平;微量酶标法测乳酸脱氢酶（lactate dehydrogenase,LDH）活力;ELISA法检测白细胞介素（interleukin,IL）?1β、IL?18的分泌水平;Western blot法检测NLRP3、凋亡相关斑点样蛋白（apoptosis?associated speck?like protein containing a CARD,ASC）、半胱氨酸天冬氨酸蛋白酶（caspase）?1、pro?caspase?1、p?NF?κBp65、NF?κBp65蛋白表达水平。结果：不同浓度的原花青素对BV2细胞存活率的影响与对照组相比差异无统计学意义（P > 0.05）。与对照组相比,LPS/ATP诱导可增加BV2细胞NO、IL?1β、IL?18水平以及LDH活力（P < 0.05）,增加NLRP3、ASC、pro?caspase?1、caspase?1、     

WP1130通过抑制NLRP3炎症小体活化缓解小鼠的感染性休克

目的 探究小分子化合物WP1130抑制NLRP3炎症小体活化并缓解感染性休克的作用机制。方法 细胞生物学水平：WP1130预处理小鼠骨髓来源巨噬细胞（BMDM）或人THP-1细胞后,利用多种NLRP3炎症小体活化剂（Nigericin、ATP、MSU、胞内LPS转染）活化NLRP3炎症小体,使用poly A:T活化AIM2炎症小体,通过Western blot和ELISA检测细胞培养上清中caspase-1和IL-1β的分泌水平,确定WP1130对NLRP3炎症小体的抑制效果及其特异性。利用激光共聚焦显微镜观察Nigericin诱导的线粒体损伤情况,探究WP1130是否影响NLRP3炎症小体组装活化的上游信号。动物水平：将SPF级雄性C57BL/6小鼠随机分为3组,即空白对照组（Control组）、感染性休克组（LPS组）、WP1130治疗组（WP1130+LPS组）,ELISA检测小鼠血清和腹腔液中IL-1β、TNF-α的分泌水平。结果 WP1130可剂量依赖性的抑制多种激动剂诱导的NLRP3炎症小体活化（P<0.05）,但对非炎症小体相关炎性因子IL-6、TNF-α的分泌无显著影响（P>0.05）。此外,WP1130对AIM2炎症小体的活化无显著影响（P>0.05）。机制研究表明,WP1130并不影响NLRP3炎症小体组装活化的上游信号线粒体损伤。动物实验结果表明,相较感染性休克组（LPS组）,WP1130治疗组（WP1130+LPS组）小鼠血清和腹腔液中IL-1β的水平显著降低（P<0.05）,而TNF-α的分泌水平无统计学差异（P>0.05）。结论 WP1130能够特异性抑制NLRP3炎症小体活化并缓解LPS诱导的小鼠感染性休克。     

多索茶碱对脂多糖诱导肺泡上皮细胞炎性损伤及NLRP3/IL-1β通路的影响

目的探讨多索茶碱对脂多糖诱导肺泡上皮细胞炎症损伤及NOD样受体蛋白3(NLRP3)/白细胞介素-1β(IL-1β)通路的影响。方法体外培养人肺泡上皮细胞(AEC),将细胞分为对照组(不做处理)、LPS组(10μg/mL LPS处理)、LPS+低剂量多索茶碱组(10μg/mL LPS+5 mg/L多索茶碱处理)、LPS+中剂量多索茶碱组(10μg/mL LPS+10 mg/L多索茶碱处理)、LPS+高剂量多索茶碱组(10μg/mL LPS+30 mg/L多索茶碱处理)。溴脱氧尿苷(BrdU)掺入实验检测细胞增殖比例,流式细胞仪检测细胞凋亡;蛋白印迹(WB)法检测各组细胞的IL-1β、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、NLRP3、凋亡相关斑点蛋白(ASC)、半胱天冬酶-1(caspase-1)蛋白表达,用凝胶电泳迁移率转变分析(EMSA)检测各组细胞核转录因子-κB(NF-κB)活性。结果与对照组比较,LPS组细胞增殖比例均下降,细胞凋亡率、细胞中IL-1β、IL-6、TNF-α、NLRP3、ASC、caspase-1蛋白表达及NF-κB活性均升高,差异均有统计学意义(P0.05);与LPS组比较,LPS+低剂量多索茶碱组、LPS+中剂量多索茶碱组、LPS+高剂量多索茶碱组细胞增殖比例依次升高,细胞凋亡率、细胞中IL-1β、IL-6、TNF-α、NLRP3、ASC、caspase-1蛋白表达及NF-κB活性均依次降低,差异均有统计学意义(P0.05),且呈剂量依赖性。结论多索茶碱可能通过抑制NLRP3/IL-1β通路活化,减轻脂多糖诱导肺泡上皮细胞炎性损伤。     

CsNOSIP与NLRP3炎症小体活化的关系及其在华支睾吸虫慢性感染过程中的作用

目的探讨华支睾吸虫一氧化氮合酶相互作用蛋白(CsNOSIP)与Nod样受体蛋白3(NLRP3)炎症小体活化的关系及其在华支睾吸虫慢性感染过程中的作用。方法脂多糖(LPS)和三磷酸腺苷(ATP)共同刺激构建NLRP3炎症小体活化人巨噬细胞(THP-1)细胞模型,荧光定量PCR(qRT-PCR)检测不同浓度CsNOSIP作用下,NLRP3炎症小体相关指标NLRP3和白细胞介素-1b(IL-1b)的表达情况;qRT-PCR法检测CsNOSIP直接刺激人肝星状细胞LX-2后肝纤维化相关指标的表达情况;CsNOSIP与华支睾吸虫溶血磷脂酶(CsLysoPLA)共孵育后,qRT-PCR法检测小鼠巨噬细胞RAW264.7白细胞介素-25(IL-25)的分泌情况;构建RAW264.7炎症细胞模型,qRT-PCR法检测肿瘤坏死因子-α(TNF-α)的表达情况。结果 qRT-PCR法检测结果显示,低浓度的CsNOSIP可抑制THP-1细胞模型内NLPR3炎症小体相关指标NLRP3和IL-1b的表达,与阳性对照组相比差异有统计学意义(P0.05)。并且CsNOSIP还能下调CsLysoPLA诱导的IL-25的表达,与单独CsLysoPLA相比,差异有统计学意义(P0.05)。CsNOSIP还可抑制LPS诱导的TNF-α的表达(F=18.71,P0.05)。结论 CsNOSIP可抑制NLRP3炎症小体活化及下调IL-25和TNF-α的表达,可能在维持华支睾吸虫慢性感染的过程中发挥着重要的作用。     

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…