内脏脂肪素对巨噬细胞MMP-9的作用及其机制探讨

目的 研究内脏脂肪素（visfatin）对巨噬细胞基质金属蛋白酶-9（MMP-9）的作用及其机制。方法体外诱导THP-1单核细胞转化为巨噬细胞。为明确visfatin对MMP-9的作用,细胞分为两组：①巨噬细胞+visfatin 12 h组;②巨噬细胞+visfatin24 h组,两组的visfatin的质量浓度均为：0（对照组）、50、100、200、400 ng/mL。采用RT-PCR和Western blotting测定MMP-9基因和蛋白表达,明胶酶谱法检测MMP-9的活性。为明确visfatin对MMP-9的作用机制,细胞分为五组：①巨噬细胞未加刺激组（对照组）;②巨噬细胞+ MAPK p38、ERK1/2、JNK信号通路抑制剂预处理1 h后加visfatin（200 ng/mL）24 h组;③巨噬细胞+过氧化物酶体增殖剂活化受体（PPARγ）天然及人工配体/ RXR配体预处理1 h后加visfatin（200 ng/mL）24 h组;④巨噬细胞+visfatin（200 ng/mL）24 h组（Vis200组）;⑤巨噬细胞+visfatin（200 ng/mL）刺激不同时间组（5、10、15、30、60 min）。Western blotting检测MMP-9蛋白和PPARγ蛋白表达及visfatin刺激下巨噬细胞p38、ERK1/2、JNK MAPK磷酸化水平。结果 Visfatin能促进MMP-9基因及蛋白表达（P<0.05,P<0.01）,同时增强了MMP-9的活性（P<0.01）。p38 MAPK、ERK1/2 MAPK通路抑制剂及RXR配体抑制visfatin对MMP-9表达具有上调作用;visfatin能促进p38 MAPK和ERK1/2 MAPK的磷酸化,但不影响PPARγ蛋白的表达。结论 Visfatin增加了巨噬细胞炎症因子的表达,该作用与p38 MAPK和ERK1/2MAPK信号通路有关;RXR可能参与了该过程。     

HB-EGF对HSC-T6细胞Ras/ERK通路的激活及对MMP-2、TIMP-1表达的影响

目的 观察肝素结合性表皮生长因子样生长因子(HB-EGF)激活肝星状细胞(HSC)过程中Ras/ERK信号通路的变化和对金属基质蛋白酶(MMP)及其抑制物的影响. 方法 传代培养HSC-T6细胞,分空白对照组、HB-EGF处理组(20,40,80 ng/mL),采用QRT-PCR法检测MMP-2、金属蛋白酶组织抑制剂-1(TIMP-1)及表皮生长因子受体(EGFR) mRNA的表达,并确定HB-EGF刺激HSC-T6 Ras/ERK通路(ERK1、ERK2、P38MAPK)活化的最佳浓度及最佳时间,检测该通路活化情况. 结果 QRT PCR检测提示,MMP-2的表达随HB-EGF干预时间的延长增加,TIMP-1的表达随干预时间的延长递减;40 ng/mL HB-EGF培养HSC-T6 48 h后,EGFRmRNA的表达较空白对照组明显上调(P＜0.05);选择40 ng/mL HB-EGF培养48 h为最佳刺激时间与浓度,该处理组ERK1 mRNA的表达较空白对照组明显上调(P＜0.05),但ERK2、P38MAPK的变化无统计学意义;Western-blot检测提示该处理组ERK1蛋白的表达较空白对照组上调(P＜0.05). 结论 HB-EGF可影响HSC-T6的MMP-2、TIMP-1的表达;HB-EGF通过促进HSC-T6表达EGFR、ERK1,有效激活细胞内Ras/ERK信号通路.     

水飞蓟宾对宫颈癌HeLa细胞增殖和侵袭能力的影响及机制研究

目的 研究水飞蓟宾(silibinin, SB)对宫颈癌HeLa细胞增殖和侵袭能力的影响,并探究其作用机制。方法 CCK-8检测不同浓度SB对宫颈癌HeLa细胞增殖的影响,筛选SB作用浓度。将细胞分为9组：对照组、SB-L(水飞蓟宾低剂量)组、SB-M(水飞蓟宾中剂量)组、SB-H(水飞蓟宾高剂量)组、PD98059(ERK1/2信号抑制剂)组、SB202190(p38 MAPK抑制剂)组、PD98059+SB组、SB202190+SB组和顺铂组;CCK-8检测细胞增殖能力,Transwell观察各组细胞侵袭情况,实时PCR检测基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)、MMP-9、细胞外信号调节激酶1/2(extracellular signal-regulated kinase 1/2,ERK 1/2)和p38 mRNA表达水平,Western blot检测MMP-2、MMP-9、ERK1/2、p38、p-ERK1/2和p-p38蛋白表达水平。结果 与对照组比较,其余各组细胞的增殖率和侵袭细胞数均显著降低(均P<0.05),MMP-...     

丝裂原活化蛋白激酶对佛波酯诱导滋养细胞MMP-9基因表达的影响

目的 探讨佛波酯(PMA)诱导细胞滋养层细胞(CTB)MMP-9基因表达的调控机制。方法 用细胞ELISA法测定CTB细胞的蛋白激酶活性变化;用反转录聚合酶链反应检测CTB中MMP-9的基因表达。结果 100 nmol/L PMA能迅速激活CTB中丝裂原活化蛋白激酶(MAPK)家族中细胞外信号调节蛋白激酶(ERK)、c-jun氨基末端激酶(JNK)以及p38 MAPK激酶的活性。100 nmol/L PMA刺激CTB引起MMP-9 mRNA表达显著增加,能被ERK或p38 MAPK的特异性抑制剂所抑制。结论 ERK和p38 MAPK可能是PMA诱导CTB中MMP-9基因表达增加的重要调节物质。     

丝裂原活化蛋白激酶对佛波酯诱导滋养细胞MMP-9基因表达的影响

目的 探讨佛波酯(PMA)诱导细胞滋养层细胞(CTB)MMP-9基因表达的调控机制。方法 用细胞ELISA法测定CTB细胞的蛋白激酶活性变化；用反转录聚合酶链反应检测CTB中MMP-9的基因表达。结果 100nmol／L PMA能迅速激活CTB中丝裂原活化蛋白激酶(MAPK)家族中细胞外信号调节蛋白激酶(ERK)、c-jun氨基末端激酶(JNK)以及p38 MAPK激酶的活性。100nmol／L PMA刺激CTB引起MMP-9 mRNA表达显著增加，能被ERK或p38 MAPK的特异性抑制剂所抑制。结论 ERK和p38 MAPK可能是PMA诱导CTB中MMP-9基因表达增加的重要调节物质。     

TWEAK通过P38MAPK途径促进大鼠心肌成纤维细胞Ⅰ型胶原和MMP-1的表达

目的 探讨肿瘤坏死因子样凋亡微弱诱导剂(TWEAK)影响心肌成纤维细胞(CFs)中Ⅰ型胶原和基质金属蛋白酶1(MMP-1)表达的作用机制。方法 取Wistar新生大鼠CFs,加入重组人TWEAK(rhTWEAK)及P38MAPK抑制剂(SB203580)干预,将实验分为① 对照组：不加干预因素;② TWEAK组：加入100ng/mL TWEAK;③ TWEAK+SB203580组：加入100ng/mLTWEAK+SB203580。 采用MTT法检测CFs增殖;Western blot法检测Ⅰ型胶原蛋白及磷酸化P38MAPK(P-P38MAPK)蛋白表达;qRT-PCR法检测Ⅰ型胶原mRNA表达;ELISA法检测CFs细胞培养液中MMP-1的浓度。结果 加入TWEAK干预后,促进了CFs增殖,Ⅰ型胶原蛋白及P-P38MAPK蛋白表达上调,Ⅰ型胶原mRNA和MMP-1 表达上调。加入SB203580可抑制CFs增殖,部分抑制Ⅰ型胶原mRNA、Ⅰ型胶原蛋白、P-P38MAPK蛋白及MMP-1的表达。结论 TWEAK可通过P38MAPK途径促进CFs 表达Ⅰ型胶原和MMP 1。     

刺糖多糖对小鼠腹腔巨噬细胞核转录因子、丝裂原活化蛋白激酶及氨基末端激酶信号通路相关基因和蛋白表达的影响

目的探讨刺糖多糖对小鼠腹腔巨噬细胞中核转录因子(NF-κB)、丝裂原活化蛋白激酶(MAPK)及cJun氨基末端激酶(JNK)信号通路相关基因和蛋白表达水平的影响。方法将处于对数生长期的小鼠腹腔巨噬细胞RAW264. 7按1×108L-1接种至24孔细胞培养板中,24 h后更换细胞培养液。将细胞分为0. 0、12. 5、50. 0、100. 0 mg·L-1刺糖多糖组,每组细胞加入相应浓度的刺糖多糖干预24 h。采用实时荧光定量聚合酶链反应检测各组细胞中p38、NF-κB p65、细胞外调节蛋白激酶1/2(ERK1/2)、JNK1/2/3 mRNA的表达,采用Western blot法检测各组细胞中磷酸化p38 (p-p38)、p38、磷酸化NF-κB p65 (p-NF-κB p65)、NF-κB p65、磷酸化ERK1/ERK2 (p-ERK1/ERK2)、ERK1/ERK2、磷酸化JNK1/2/3(p-JNK1/2/3)及JNK1/2/3蛋白的表达。结果刺糖多糖干预巨噬细胞24 h后,与0. 0 mg·L-1刺糖多糖组比较,50. 0、100. 0 mg·L-1刺糖多糖组细胞中ERK1、NF-κB p65 mRNA表达量显著升高(P <0. 05); 12. 5、50. 0、100. 0 mg·L-1刺糖多糖组细胞中p38 mRNA表达量显著升高(P <0. 05)。与12. 5 mg·L-1刺糖多糖组比较,50. 0、100. 0 mg·L-1刺糖多糖组细胞中ERK1、p38、NF-κB p65 mRNA表达量显著升高(P <0. 05)。与0. 0、12. 5 mg·L-1刺糖多糖组比较,50. 0、100. 0 mg·L-1刺糖多糖组细胞中p-p38、p-NF-κB p65、p-ERK1/ERK2、ERK1/ERK2蛋白表达量显著升高(P <0. 05)。结论刺糖多糖能上调p38、NF-κB p65、ERK1/2蛋白的磷酸化水平,从而激活MAPK信号通路和NF-κB信号通路;刺糖多糖的免疫机制可能与NF-κB p65、p38、ERK1mRNA表达上调有关。     

长链非编码RNA核富集转录子1对胃癌SGC7901细胞迁移、侵袭、自噬的影响及机制研究

目的:分析长链非编码RNA(lncRNA)核富集转录子1(NEAT1)通过调控丝裂原活化蛋白激酶(MAPK)信号通路对胃癌SGC7901细胞迁移、侵袭及自噬的影响。方法:人胃癌SGC7901细胞传代培养后分为对照组、过表达组和沉默组。对照组为不做任何处理，过表达组给予lncRNA NEAT1过表达质粒转染，沉默组给予lncRNA NEAT1沉默质粒转染。鉴定lncRNA NEAT1转染效率，分析沉默lncRNA NEAT1对人胃癌SGC7901细胞迁移、侵袭、自噬、MAPK信号通路蛋白表达量的影响。结果:与对照组比较，过表达组lncRNA NEAT1表达量、基质金属蛋白酶-9(MMP-9)、MMP-2、细胞外信号调节激酶(ERK)蛋白表达量增加，上皮性钙黏附素(E-cadherin)、Beclin-1、Ⅱ型/Ⅰ型微管相关蛋白1轻链3(LC3-Ⅱ/LC3-Ⅰ)、p38、c-Jun氨基末端激酶(JNK)蛋白表达量降低；沉默组lncRNA NEAT1表达量、MMP-9、MMP-2、ERK蛋白表达量降低，E-cadherin、Beclin-1、LC3-Ⅱ/LC3-Ⅰ、p38、JNK蛋白表达量增...     

过氧化物酶体增殖体活化受体γ、肝脏X受体α、视黄酸X受体α活化对THP-1单核巨噬细胞ABC1mRNA表达的影响及意义

目的探讨核受体过氧化物酶体增殖剂活化受体γ(peroxisome proliferator-activated receptorγ,PPARγ)、肝脏X受体α(Liver X receptor alpha,LXRα)、视黄酸X受体α(Retinoid X receptor alpha,RXRα)活化对THP-1单核巨噬细胞ATP结合盒转运蛋白(ATP binding cassette transporter 1,ABC1)mRNA表达的影响及意义。方法体外培养的THP-1单核细胞,经佛波酯(PMA)诱导成为巨噬细胞后,与核受体PPARγ、LXRα、RXRα的相应配体曲格列酮(TROG)、22(R)羟化胆固醇(22(R)-HC)、9-顺式视黄酸(9-CRA)作用24小时,提取细胞总RNA,用一步法逆转录聚合酶链反应检测核受体PPARγ、LXRα、RXRα活化剂对ABC1mRNA表达的影响。结果与对照组相比较,曲格列酮组、22(R)-HC组、9-CRA组、TROG+22(R)-HC组、TROG+组、22(R)-HC+9-CRA组ABC1 mRNA的表达均有不同程度增加,联合作用组(即核受体PPARγ、LXRα、RXRα两两活化)ABC1 mRNA增加更为明显。结论通过调控PPARγ、LXRα和RXRα等核受体的活化程度可达到调控ABC1表达的目的,这可能将为动脉粥样硬化的防治提供新的前景和机会。     

p38MAPK对大鼠肾缺血再灌注损伤早期MMP-9表达的调控

目的 探讨p38MAPK对大鼠肾缺血再灌注后MMP-9表达的调控情况及它们在肾缺血再灌注损伤中的作用.方法 48只健康雄性SD大鼠,分组建立大鼠肾缺血再灌注损伤模型,测定各组实验大鼠血清肌酐水平的变化,应用免疫组化二步法检测p38MAPK、MMP-9表达的变化.结果 与假手术组比较,缺血再灌注各组血清肌酐水平升高,缺血45min再灌注,24h达高峰(P＜0.01).MMP-9蛋白在假手术组呈少量散在表达或不表达,缺血45min再灌注6h有少量表达,再灌注12、24h及72h呈阳性表达,在24h达高峰(P＜0.01);p38MAPK蛋白在假手术组呈少量散在表达或不表达,缺血45 min,再灌注6、12h及24 h呈阳性表达,12h达高峰(P＜0.01).p38MAPK,MMP-9蛋白的表达与血清肌酐水平的变化呈显著正相关.结论 肾缺血再灌注可激活p38MAPK,活化的p38MAPK可以上调MMP-9蛋白的表达,可能促进了肾缺血再灌注损伤的发生和发展.     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

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…     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

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.     

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.     

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.     

The clinicopathological and immuohistochemical analysis of solid-pseudopapillary tumor of the pancreas： report of 9 cases

Objective：To investigate the clinical features, pathological characteristics and immunophenotype of solid-pseudopapillary tumor of the pancreas（SPTP）. Methods：Nine surgically treated cases of SPTP were retrospectively reviewed. Hematoxylin and Eosin（HE） staining and immunohistochemical staining were used to analyze all cases, and the general clinical data was collected. Results：Six patients were asymptomatic except for a palpable mass. Two patients complained of vague-epigastric pain. One patient appeared jaundice. The tumor was encapsulated and solid tissues alternately with cystic tissues. Histologically, the histological structure of solid portion was pseudopapillary with a fibrovascular core. Tumor cells were uniform and medium-sized which were arranged in sheets ets or nests or pseudopapillary patterns. Immunohistochemical studies demonstrated that SPTP proved positive in vimentin（9/9 cases）, AAT（9/9 cases）, NSE（9/9 cases）, ACT（7/9 cases）, CK20（2/9 cases）, CgA（1/9 cases）, S-100（3/gcases）, PR（4/gcases）, Syn（3/9 cases） and CD56（5/9cases）, negative in CEA and ER. Conclusion：SPTP is a tumor predominantly occurring in young women frequently without special symptoms. This tumor has various characteristical histological patterns with different immunophenotype.