NLRP3炎症小体及其下游炎症因子在动脉粥样硬化炎症反应中的作用

目的：探讨NLRP3炎症小体及其下游炎症因子在动脉粥样硬化炎症反应中的作用。方法选择氧化低密度脂蛋白(ox-LDL)作用于人单核细胞系THP-1,经Western blot法检测NLRP3及其下游炎症因子半胱天冬酶-1(Caspase-1)、白介素-1β(IL-1β)和白介素-18(IL-18)的表达;并采用SiRNA技术下调空白处理组(Blank组)和ox-LDL处理组NLRP3基因表达,分别检测以上指标。结果与空白处理组相比,50 mg/L、100 mg/L、150 mg/L的ox-LDL作用于单核细胞后可促进NLRP3的表达,NLRP3下游的Caspase-1、IL-1β和IL-18表达水平与NLRP3表达一致,表达也均明显升高,差异均具有统计学意义(P<0.05)。NLRP3小干扰RNA (SC-45469)转染后,Blank组与ox-LDL组的NLRP3及其下游炎症因子Caspase-1、IL-1β和IL-18的表达均有显著下降(P<0.05);而转染后Blank组与ox-LDL组各指标下降率组间比较差异均无统计学意义(P>0.05)。结论 NLRP3炎症小体参与动脉粥样硬化发生发展的过程,其可能通过改变ox-LDL的表达水平,诱发炎症相关因子Caspase-1、IL-1β和IL-18的分泌来调节, NLRP3炎症小体具有成为临床上防治动脉粥样硬化的新靶点的重要潜力。     

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。     

卡维地洛对ox-LDL诱导内皮细胞中NLRP3炎症小体介导焦亡及炎症的影响

目的研究卡维地洛对氧化型低密度脂蛋白(ox-LDL)诱导内皮细胞中NLRP3炎症小体介导焦亡及炎症的调控作用。方法人脐静脉内皮细胞(HUVEC)分为对照组、ox-LDL组、低剂量组(0.01 mmol/L)、高剂量组(0.1 mmol/L);转染阴性对照腺病毒(Ad-NC)或过表达NLRP3腺病毒(Ad-NLRP3)后分为Ad-NC组、Ad-NC+ox-LDL组、Ad-NC+高剂量组、Ad-NLRP3+高剂量组。检测各组细胞活力和白细胞介素(IL)-1β、IL-18含量,以及GSDMD-N、NLRP3、Cleaved Caspase-1表达水平。 结果ox-LDL组细胞活力低于对照组,IL-1β、IL-18及GSDMD-N、NLRP3、Cleaved Caspase-1均高于对照组(P＜0.05)。低剂量组和高剂量组细胞活力高于ox-LDL组,IL-1β、IL-18及GSDMD-N、NLRP3、Cleaved Caspase-1均低于ox-LDL组(P＜0.05)。转染NLRP3腺病毒后,Ad-NLRP3+高剂量组和Ad-NC+ox-LDL组细胞活力低于Ad-NC+高剂量组,IL-1β、IL-18及GSDMD-N、NLRP3、Cleaved Caspase-1均高于Ad-NC+高剂量组(P＜0.05)。 结论卡维地洛显著抑制ox-LDL诱导HUVEC中NLRP3炎症小体介导的细胞焦亡及炎症反应。     

游离脂肪酸激活肾系膜细胞NLRP3炎症小体信号及氧化应激机制的探讨

目的:探讨NLRP3炎症小体在游离脂肪酸(free fatty acids,FFAs)诱导下的肾小球系膜细胞(HBZY-1)中的表达及氧化应激在其中的影响。方法:将HBZY-1细胞分为正常对照组、不同浓度棕榈酸组、高棕榈酸+N-乙酰半胱氨酸(N-acetylL-cysteine,NAC)组。各组细胞培养6、9、12 h后采用Western blotting和RT-PCR检测各组NLRP3、Caspase-1、IL-1β蛋白及m RNA表达,并同时观察NAC对各组蛋白及m RNA表达的影响。结果:棕榈酸刺激肾系膜细胞NLRP3炎症小体信号分子的表达变化:与对照组相比,不同作用浓度和时间的棕榈酸均可上调NLRP3、Caspase-1、IL-1β蛋白及m RNA表达(P<0.05),且呈浓度和时间依赖性。与单纯棕榈酸组相比,NAC预处理可阻止棕榈酸诱导的NLRP3、Caspase-1、IL-1β蛋白及m RNA表达上调(P<0.05)。结论:棕榈酸可通过氧化应激机制诱导系膜细胞NLRP3炎症小体信号的活化,参与糖尿病肾病炎症的发生。     

NLRP3炎症小体在脓毒症及其相关器官功能障碍中的研究进展

脓毒症是一个复杂而动态的过程,与免疫反应密切相关.炎症小体被认为是脓毒症固有免疫反应的重要组成部分,其中以核苷酸结合寡聚化结构域样受体蛋白3(nucleotide-binding oligomerization domain-like receptors family pyrin domain containing 3,NLRP3)炎症小体研究最广泛,它能与接头蛋白ASC、Caspase-1结合后形成炎症小体复合体,通过调控Caspase-1活化,促进白细胞介素-1β(interleukin-1β,IL-1β)及白细胞介素-18(interleukin-18,IL-18)等炎症因子成熟、释放.本文就近年来NLRP3炎症小体在脓毒症及其相关器官功能障碍中的研究进展进行综述.     

Mcc950在新生儿坏死性小肠结肠炎的作用及其机制研究

目的 研究NLRP3炎症小体及其特异性抑制剂Mcc950对新生儿坏死性小肠结肠炎(NEC)的作用及机制.方法 将120只1日龄新生SD乳鼠随机分为对照组、NEC组和Mcc950治疗组,每组40只.通过人工喂养、缺氧复氧、冷刺激加脂多糖(LPS)灌胃建立NEC动物模型.采用HE染色观察各组肠组织病理表现,并对肠组织行损伤评分;采用蛋白质印迹技术和实时荧光定量PCR(RT-qPCR)检测炎症因子IL-1β和IL-18、炎症小体NLRP3、细胞焦亡主要标志分子Caspase-1和GSDMD的蛋白及mRNA表达水平.结果 与对照组相比,NEC组大鼠体重减轻,肠上皮细胞排列紊乱,部分绒毛坏死、脱落,黏膜层、黏膜下层及固有层可见水肿分离,相应的肠组织损伤评分升高,IL-lβ、IL-18、NLRP3、Caspase-1和GSDMD蛋白及mRNA表达水平升高,差异有统计学意义(P＜0.05).与NEC组相比,Mcc950治疗组大鼠体重下降趋势变缓,肠组织病理损伤减轻,肠组织损伤评分降低,IL-1β、IL18、NLRP3、Caspase-1和GSDMD表达降低,差异有统计学意义(P＜0.05).结论 成功建立NEC模型,Mcc950可抑制NLRP3介导的炎症及细胞焦亡,从而在NEC中发挥保护作用.     

NLRP3与NOD2受体激活对下游炎症因子表达的影响

目的 核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)与核苷酸结合寡聚化结构域2(NOD2)受体被其特异性配体激活后对下游炎症因子表达的影响.方法 实验分为4组:空白对照组、TATP组、TMDP、TMDP+ATP组.用酶联免疫吸附试验(ELISA)测定白细胞介素(IL)-1β及IL-18的表达,逆转录PCR测定NLRP3、半胱氨酸天冬氨酸蛋白酶1(Caspase-1)的表达情况.结果 TMDP组、TATP组、TMDP+ ATP组的IL-1β和IL-18水平均高于空白对照组(P＜0.05),且TMDP+ ATP组水平增高更显著.TMDP组、TATP组、TMDP+ATP组的NLRP3和Caspase-1表达均高于空白对照组(P＜0.05),且TMDP+ATP2水平增高更显著(P＜0.05).结论 NOD2的特异性配体胞壁酰二肽,NLRP3的特异性配体腺苷三磷酸单独刺激THP-1后NLRP3及下游炎症因子的表达很弱或无表达,联合刺激后可显著增强NLRP3及下游炎症因子的表达,二者之间存在相互促进及协同作用.     

NLRP3炎症小体在高糖诱导小鼠肾小管上皮细胞EMT中的作用

目的:观察NLRP3炎症小体在高糖诱导小鼠肾小管上皮细胞上皮间质转化(EMT)中的作用.方法:以小鼠肾小管上皮细胞为研究对象,高糖处理24 h后,采用real-time PCR法检测细胞表型标志分子(E钙粘素和α-SMA)和NLRP3炎症小体mRNA的表达,ELISA法检测细胞上清液中IL-1β含量.采用NLRP3炎症小体特异性抑制剂MCC950探讨NLRP3炎症小体在高糖诱导小鼠肾小管上皮细胞ETM中的作用.结果:高糖处理24 h可降低小鼠肾小管上皮细胞E钙粘素mRNA表达,上调α-SMA mRNA表达;同时上调NLRP3、pro-IL-1β和pro-caspase-1 mRNA表达,以及IL-1β分泌;抑制NLRP3炎症小体可部分逆转高糖诱导的小鼠肾小管上皮细胞EMT.结论:高糖应激可能通过激活NLRP3炎症小体促进IL-1β分泌诱导肾小管上皮细胞发生EMT.     

左旋紫草素对大鼠重症急性胰腺炎并发急性心肌损伤的保护作用研究

目的 探讨左旋紫草素对大鼠重症急性胰腺炎(SAP)并发急性心肌损伤(AMI)的保护作用.方法 24只雄性SD大鼠随机分为假手术组、SAP模型组与左旋紫草素干预组(20 mg/kg),每组8只.采用L-精氨酸腹腔注射制备SAP模型,并于术后24 h内腹腔注射左旋紫草素进行干预治疗.HE染色观察胰腺及心肌组织病理变化;酶联免疫吸附剂测定法测定各组血清肌酸激酶同工酶(CK-MB)、肿瘤坏死因子(TNF-α)、白介素1β(IL-1β)水平;蛋白质印迹技术检测各组心肌半胱氨酸天冬氨酸蛋白酶(Caspase-1)、炎性小体(NLRP3)、凋亡相关斑点样蛋白(ASC)蛋白表达;DHE荧光探针法检测各组心肌活性氧ROS含量.结果 与假手术组相比,SAP模型组大鼠胰腺和心肌组织损伤严重,血清CK-MB、TNF-α、IL-1β水平增加,Caspase-1、NLRP3、ASC蛋白表达水平增高,心肌ROS水平增加,差异均有统计学意义(P<0.05).与SAP模型组比较,左旋紫草素干预组大鼠胰腺和心肌组织损伤程度得到改善,血清CK-MB、TNF-α、IL-1β水平降低,Caspase-1、NLRP3、ASC蛋白表达水平减少,心肌ROS水平降低,差异均有统计学意义(P<0.05).结论 左旋紫草素可减轻SAP并发的AMI,其机制可能与抑制心肌组织NLRP3炎性小体过度活化有关.     

慢性牙周炎牙龈卟啉单胞菌感染患者NLRP3炎症小体与炎症反应及焦亡指标的相关性

目的研究慢性牙周炎牙龈卟啉单胞菌感染患者NLRP3炎症小体与炎症反应及焦亡指标的相关性。 方法选择300例慢性牙周炎患者,按照牙龈卟啉单胞菌感染情况分为阳性组和阴性组。比较两组牙周健康指标、牙周组织NLRP3、凋亡相关斑点样蛋白(ASC)、Caspase-1 mRNA和GSDMD-N蛋白表达水平,以及龈沟液白细胞介素(IL)-1β、IL-18、肿瘤坏死因子-α(TNF-α)水平。采用Pearson检验进行相关性分析。 结果阳性组菌斑指数(PLI)、牙龈指数(GI)、附着丧失(AL)、牙周袋深度(PD)、牙周组织NLRP3、ASC、Caspase-1 mRNA、GSDMD-N蛋白,以及龈沟液IL-1β、IL-18、TNF-α水平均高于阴性组(P＜0.05)。NLRP3、ASC、Caspase-1 mRNA表达水平与PLI、GI、AL、PD、GSDMD-N蛋白、IL-1β、IL-18、TNF-α水平均呈正相关(P＜0.05)。 结论慢性牙周炎患者牙龈卟啉单胞菌感染加重病情、炎症反应及细胞焦亡,其作用可能与激活NLRP3炎症小体有关。     

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.     

Acupuncture Treatment of Vomiting

CASE HISTORY A female patient, 46 years old, head of the foreign affairs department of a certain university in Beijing, paid her first visit on October 9, 2006, with the chief complaint of vomiting for one month. She got vomiting after meals in early September. Before that, she had discomfortable sensation in the stomach due to angry with others, but she didn't pay much attention. Later, it developed into vomiting after eating. After the vomiting, the discomfort would be relieved, but with slight hypodynamia. She was once diagnosed as having 'neurogenic vomiting'. Having taken some western and Chinese drugs, the above symptoms were a little bit improved, but she would have nausea upon eating and with regurgitation. Because of the fear for vomiting, she did not dare to have food intake, with body weight reduction of 6 kilos in one month.     

Qiu Baoguo''''s Experience in Treating Consumptive Syndromes after Radio-Chemotherapies

Radiotherapy and chemotherapy are the important modern medical therapies for malignant tumors,yet they can also bring about serious local and systemic toxic side reactions so to decrease the patient;'s life quality,manifested by a series of consumptive symptoms.Having engaged in the combined work of Chinese and western medicine for nearly 50 years,the research fellow Qiu Baoguo in Henan Provincial Academy of TCM has developed his unique views on the TCM study of consumptive syndromes.The author of this essay had once the fortune tO follow Dr.Qiu in clinic,and specially would like to introduce in the following Dr.Qiu's experience in treating consumptive syndromes after radio-chemotherapies for patients with malignant tumor.     

Insomnia Due to Deficiency of Both the Heart and Spleen Treated by Acupuncture-Moxibustion and Chinese Tuina

OBJECTIVE: To observe therapeutic effects of the comprehensive therapy of acupuncture-moxibustion and Chinese Tuina for treatment of insomnia due to deficiency of both the heart and spleen. METHODS: 92 cases were divided randomly into the treatment group (treated by acupuncture-moxibustion and Chinese Tuina) and the control group (treated by acupuncture-moxibustion). RESULTS: The therapeutic effect of the treatment group was obviously superior to that of the control group (the CHI2 test showed P < 0.01). CONCLUSIONS: The comprehensive therapy of acupuncture-moxibustion and Chinese Tuina can give marked therapeutic effects for treatment of insomnia due to deficiency of both the heart and spleen.     

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.