青藤碱对佐剂性关节炎大鼠TLR4/MyD88信号通路的影响

目的:探讨青藤碱对佐剂性关节炎大鼠TLR4/MyD88信号通路诱导炎症的影响。方法:选择清洁级SD大鼠40只,随机分为模型组、青藤碱组、甲氨蝶呤组及正常对照组。前3组大鼠右后足跖皮内注射弗氏完全佐剂诱发大鼠佐剂性关节炎模型,正常对照组大鼠右后足跖皮内注射等量0.9%氯化钠注射液。4组分别干预4周后,取膝关节滑膜,采用Real-time PCR法检测TLR4 mRNA、MyD88 mRNA表达,ELISA法检测炎性因子TNF-α。结果:模型组大鼠膝关节滑膜组织TLR4 mRNA、MyD88 mRNA及炎性因子TNF-α表达均较正常对照组明显升高(P<0.01)。经青藤碱治疗后,TLR4 mRNA、MyD88 mRNA及炎性因子TNF-α表达均较模型组明显下降(P<0.01)。结论:青藤碱可改善佐剂性关节炎大鼠的关节炎症,该作用可能与抑制TLR4/MyD88信号通路有关。     

脓毒症早期大鼠肝脏Toll样受体4基因表达及干预性研究

目的:探讨严重腹腔感染所致脓毒症大鼠肝脏Toll样受体4(Tolllikereceptor4,TLR4)基因表达变化规律及对其调节机制进行初步探讨。方法:采用大鼠盲肠结扎穿孔(CLP)模型造成脓毒症。将96只大鼠随机分为正常对照组、假手术组、CLP组、地塞米松(DXM)干预组,于CLP后2、4、6、12、24h处死动物,留取肝脏标本,以半定量逆转录多聚酶链反应技术及相关软件分析不同组TLR4mRNA、TNF鄄αmRNA的表达,检测血清丙氨酸转移酶(ALT)、天冬氨酸转氨酶(AST)及肝脏组织病理,评估肝脏损伤情况。结果:CLP组2h后肝脏TLR4mRNA、TNF鄄αmRNA及血清ALT、AST比正常对照组明显增多(P﹤0.01);DXM干预组各时间点TLR4mRNA、TNF鄄αmRNA的表达及血清ALT、AST较单纯CLP组显著降低(P﹤0.01),24h病理切片显示肝损伤轻于CLP组。结论:严重腹腔感染可致大鼠肝脏TLR4mRNA持续高表达,与肝脏的损伤密切相关;早期应用DXM可抑制TLR4mRNA的表达,减轻肝脏损伤。DXM抑制腹腔感染所致的炎症反应可能与抑制TLR4的表达有关。     

γ干扰素对实验性肝纤维化大鼠Smad3 mRNA的影响

目的初步探讨γ干扰素抗纤维化的分子机制。方法SD大鼠70只,模型对照组和γ干扰素治疗组注射40?l4油剂按0.3ml/100g皮下注射,2次/周,另设正常对照组大鼠10只。γ干扰素治疗组造模同时肌内注射γ干扰素0.2MU/kg,1次/日,模型对照组及正常对照组肌内注射生理盐水,连续干预12周。第12周末处死所有大鼠。取肝脏标本行常规苏木素-伊红和胶原染色,在光镜观察肝组织炎症活动度和肝纤维化情况并进行评分;实时荧光定量PCR方法检测大鼠肝脏Smad3mRNA的表达。结果肝组织炎症活动度和肝纤维化评分显示γ干扰素治疗组大鼠炎症程度、肝纤维化程度与模型对照组大鼠的肝组织炎症程度、肝纤维化程度比较显著改善(P<0.01,P<0.01)。荧光定量RT-PCR检测显示:与正常对照组Smad3mR-NA相比,模型对照组大鼠肝脏中Smad3mRNA表达显著增加(P<0.01);与模型对照组相比,γ干扰素组大鼠肝脏组织中Smad3mRNA表达明显减少(P<0.05)。结论γ干扰素抗实验性肝纤维化作用与下调Smad3mRNA来实现。干预转化生长因子β1信号转导过程是γ干扰素抗肝纤维化的机制之一。     

wortmannin对重症急性胰腺炎大鼠肺损伤的保护作用

目的:观察磷脂酰肌醇-3激酶(PI3K)抑制剂wortmannin对重症急性胰腺炎(SAP)大鼠肺损伤的保护作用并探讨其机制.方法:将健康成年SD大鼠54只随机分为对照组、SAP组和SAP wortmannin组,每组18只,逆行胆胰管注射50 g/L牛磺胆酸钠制备SAP模型.检测血清TNF-α水平、肺组织湿/干质量比值、肺组织髓过氧化物酶(MPO)活性、支气管肺泡灌洗液(BALF)蛋白含量;观察肺组织及胰腺组织的病理变化.结果:SAP组较对照组血清TNF-α水平、肺组织湿/干质量比值、BALF蛋白含量及肺组织MPO活性均显著升高(P《0.01),胰腺、肺组织病理损伤随病情进展而逐渐加重;SAP wortmannin组较对照组各项指标均升高,但较SAP组均明显降低(P《0.01), 胰腺、肺组织病理损伤较SAP组减轻.结论:wortmannin对SAP大鼠肺损伤有一定的保护作用, 其机制可能与其抑制了中性粒细胞内PI3K信号转导通路的活化, 使多种炎症细胞的活化和TNF-α等炎症因子的释放受到抑制有关.     

高脂饮食诱导的肥胖大鼠下丘脑及肝脏组织中Toll样受体4的表达

目的 探讨在高脂饮食诱导下Toll样受体4(Toll-like receptor4,TLR4)在肥胖大鼠下丘脑及肝脏组织中的表达.方法 建立高脂饮食诱导的肥胖(DIO)大鼠模型,应用免疫组化染色和Real Time PCR方法检测下丘脑及肝脏组织中TLR4的蛋白及mRNA的表达.结果 喂养8周后,DIO组平均体重为(456.38±19.20)g,高于正常饮食对照组平均体重(372.10±22.47)g,两组间比较差异有显著性意义(P＜0.01);DIO组饮食量为(1229.23±57.29)g,高于正常饮食对照组饮食量(905.50±52.17)g,两组大鼠饮食量比较差异有显著性意义(P＜0.01).Real Time PCR方法显示,DIO组大鼠下丘脑组织TLR4 mRNA含量(0.56±0.16),高于正常饮食对照组TLR4 mRNA (0.39±0.10),P＜0.05; DIO组大鼠肝脏组织TLR4 mRNA含量(0.87±0.24),高于正常饮食对照组TLR4 mRNA(0.54±0.16),两组间差异也有显著性意义(P＜0.01).免疫组化染色肥胖组大鼠下丘脑及肝脏TLR4表达呈强阳性.结论 Toll样受体4在肥胖大鼠下丘脑及肝脏中的表达增多,提示下丘脑及肝脏的炎症反应参与了高脂饮食诱导肥胖的发生.     

腺苷蛋氨酸对过度激活的Rho-ROCK通路介导的糖尿病肝损伤的改善作用

目的探讨腺苷蛋氨酸对过度激活的Rho-ROCK通路介导的糖尿病肝损伤的改善作用。方法 40只清洁级Wistar大鼠依据随机数字表法为4组:对照组、2型糖尿病组、Y27632干预组及腺苷蛋氨酸干预组,各10只。对照组大鼠给予正常饲粮,自由饮水;糖尿病组、Y27632干预组及腺苷蛋氨酸干预组大鼠每日给予高糖高脂饲料,自由饮水,并于第35日时一次性腹腔注射链脲佐菌素(35 mg/kg,腹膜内);Y27632干预组于第9~12周给予Y27632 10 mg/kg进行治疗。腺苷蛋氨酸干预组于第9~12周给予腺苷蛋氨酸10 mg/kg进行治疗。分析各组大鼠胰岛素抵抗系数、血脂水平及Rho A、ROCK1/2和基质金属蛋白酶的表达变化。同时分析各组大鼠肝脏组织细胞凋亡相关蛋白胱天蛋白酶(caspase)-3及caspase-9的表达变化。结果糖尿病组、Y27632干预组和腺苷蛋氨酸干预组大鼠的胰岛素抵抗指数稳态模型、血糖、低密度脂蛋白胆固醇较对照组明显升高(P<0.01);糖尿病组、Y27632干预组和腺苷蛋氨酸干预组大鼠的总胆固醇高于对照组(P<0.01);糖尿病组、Y27632干预组和腺苷蛋氨酸干预组大鼠的高密度脂蛋白胆固醇低于对照组(P<0.01);糖尿病组、Y27632干预组和腺苷蛋氨酸干预组大鼠的羟脯氨酸高于对照组(P<0.01)。糖尿病组大鼠肝脏组织的Rho及ROCK1/2蛋白的表达、MMP-2及MMP-9的表达及促凋亡蛋白相关蛋白caspase-3和caspase-9的表达较对照组明显升高(P<0.01),而Y27632干预组和腺苷蛋氨酸干预组上述异常表达得到一定的恢复(P<0.01)。结论腺苷蛋氨酸通过抑制肝脏组织中过度激活的Rho/ROCK信号转导通路改善小剂量SZT合并高糖高脂饮食复制的2型糖尿病大鼠肝脏组织损伤。     

板蓝根多糖通过TLR4-NF-κB通路对结核大鼠肺部炎症影响的机制研究

目的:探究板蓝根多糖通过Toll样受体4(toll-like receptor 4,TLR4)-核因子κB(nuclear factorκB,NF-κB)通路对结核大鼠肺部炎症的影响机制。方法:45只SD大鼠随机分为3组(n=15):对照组、模型组、板蓝根多糖组。通过尾静脉注射H37RV结核分枝杆菌建模,在感染的第2天板蓝根多糖组大鼠接受板蓝根多糖灌胃干预。检测和比较各组小鼠血清炎性因子以及肺组织TLR4-NF-κB通路和炎性因子表达水平。结果:模型组肺组织中感染大量的结核分枝杆菌,并且成团聚集在细胞外,板蓝根多糖组的CFU水平显著低于模型组(P<0.05),结核分歧杆菌数量少于模型组(P<0.05),主要分布于结核结节和巨细胞中。模型组血清炎性因子水平显著高于对照组,板蓝根多糖组血清中白细胞介素质1β(IL-1β)、白细胞介素质6(IL-6)、干扰素-γ(IFN-γ)水平显著低于模型组而显著高于对照组(P<0.05)。模型组肺组织中TLR4-NF-κB通路及炎性因子mRNA水平显著高于对照组,板蓝根多糖组肺组织中TLR4、NF-κB、IL-1β、IL-6、IFN-γ水平显著低于模型组而显著高于对照组(P<0.05)。模型组肺组织中TLR4-NF-κB通路表达水平显著上调,板蓝根多糖组肺组织中TLR4、NF-κB蛋白水平显著低于模型组而显著高于对照组(P<0.05)。结论:板蓝根多糖可以通过调节TLR4-NF-κB通路减轻结核分歧杆菌感染引起的肺部炎症反应。     

甘露消毒丹对温病湿热证大鼠TLR4 mRNA、NF-κB p65表达的影响

【目的】观察甘露消毒丹对温病湿热证大鼠细胞内毒素Toll样受体4(TLR4)mRNA及核因子-κB(NF-κB)p65表达变化的影响,探讨清热化湿方的治疗机制。【方法】将Wistar大鼠随机分为正常组、湿热模型组(高脂 高温高湿 大肠杆菌)、清热化湿组;采用逆转录—聚合酶链反应技术(RT-PCR)检测肝组织巨噬细胞TLR4 mRNA,免疫组化技术检测肝组织巨噬细胞NF-κB p65表达。【结果】模型组在感染6、12、24 h不同时相点TLR4 mRNA、NF-κB p65表达逐渐增强,且不同时相点之间比较均有显著性差异(P<0.05或P<0.01),说明随着病程的发展,湿热模型TLR4 mRNA、NF-κB p65表达逐渐增强。治疗组6、12、24 h不同时相点的TLR4 mRNA表达逐渐减弱,12、24 h时相点治疗组与模型组比较,TLR4mRNA表达显著性减弱(P<0.05或P<0.01),但与正常组比较仍有显著性差异(P<0.01),说明TLR4 mRNA遏制衰减是一个缓慢的过程。而治疗组NF-κB p65激活6、12、24 h各时相点间比较均无显著性差异,与正常组比较亦无显著性差异,但与同时点模型组比较均有显著性差异(P<0.05或P<0.01)。【结论】甘露消毒丹对温病湿热证大鼠肝巨噬细胞TLR4 mRNA及NF-κB p65表达具有较好的干预调控作用,能中止炎症介质的转录,限制急性炎症反应。     

Hemin诱导大鼠肝脏HO-1表达对非酒精性脂肪性肝炎的保护作用

目的观察氯化血红素(hemin)对大鼠肝脏血红素加氧酶1(HO-1)表达的诱导作用,并探讨HO-1对大鼠非酒精性脂肪性肝炎(NASH)的保护作用。方法雄性SD大鼠随机分为对照组、模型组和干预组,每组8只。对照组给予正常饮食,模型组及干预组均给予髙脂饮食,共8周。之后对照组继续正常饮食喂养,模型组髙脂饮食喂养,干预组给予髙脂饮食及每日hemin 15 mg/kg腹腔注射,共10 d。第10天处死大鼠,观察肝脏组织形态学,检测各组血清丙二醛(MDA)和谷胱甘肽(GSH)、谷丙转氨酶(ALT)、谷草转氨酶(AST)水平,Western blot检测各组肝脏组织HO-1的表达。结果模型组MDA、AST、ALT较对照组显著升高(P<0.01),干预组MDA、AST、ALT较模型组显著降低(P<0.01);模型组GSH较对照组显著降低(P<0.01),干预组GSH较模型组显著升高(P<0.01);hemin干预组肝细胞肿胀、炎细胞浸润等形态学改变较模型组明显改善。Western blot结果显示:hemin干预组HO-1的表达明显高于模型组与对照组。结论 Hemin能够诱导大鼠肝脏组织HO-1表达的增加。通过增加HO-1的表达能够减轻大鼠肝脏氧化应激损伤,改善肝脏组织学及肝功能情况,对髙脂饮食引起的NASH起到保护作用。     

宫内生长受限大鼠肝脏Toll样受体4表达的变化

目的 检测宫内生长受限(IUGR)大鼠肝脏中Toll样受体4(TLR4)及细胞因子的表达,探讨IUGR个体革兰阴性(G-)细菌感染率增加的分子机制.方法 采用孕期蛋白质营养不良法建立IUGR大鼠模型,应用荧光定量RT-PCR技术检测3周龄子鼠肝脏中TLR4的mRNA含量变化,Western blot方法检测子鼠肝脏TLR4的蛋白表达,并应用酶联免疫吸附法(ELISA)检测肝组织中肿瘤坏死因子α(TNF-α)和白介素1β(IL-1β)的水平.结果 与对照组相比,IUGR组子鼠肝脏TLR4的mRNA和蛋白表达都明显降低(P<0.01);而TNF-α和IL-1β的表达水平明显增高,且与TLR4蛋白含量呈负相关(P<0.05).结论 宫内生长受限引起肝脏中细胞因子TNF-α和TL-1β的浓度增高,可能导致TLR4表达下降,后者可引起IUGR个体革兰阴性细菌炎症和败血症增加.     

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…     

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.     

Analysis of CK-19,PDX-1,Nestin,Ngn3 in different donor islet purity in rats

Objective: To investigate if there are the CK-19, PDX-1, Nestin, Ngn3 positive cells in the donor islets of different purity in rats. Methods: Thirty male adult SD rats were randomly divided into 3 groups. Islets were isolated using digestion by ductal injection of collagenase. Group Ⅰ (n=10): Separating cell preparations were not purified, Group Ⅱ(n=10): Islet sediment was purified with 25% Ficoll400 ,Group Ⅲ (n=10): Islet sediment was purified with 25% and 11% Ficoll-400. The levels of protein of CK-19, PDX-1, Nestin and Ngn3 were detected by immunohistochemistry and the mRNA of CK-19, PDX-1, Nestin, Ngn3 was amplified by RT-PCR. Results: After two different purification methods applied, three islet preparations of different purities were obtained. The difference of islet purity was significant among various groups (P＜0.05). Compared with group Ⅱ and group Ⅲ,the protein and mRNA of CK-19, PDX-1, Nestin,Ngn3 were both higher in group Ⅰ; group Ⅲ was poorly expressed. Conclusions: The three different islet purity donor islet have different CK-19, PDX-1, Nestin, Ngn3 positive cells within them, indicating that there are some islet stem cells in the purified donor islet.     

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.