核因子-κB抑制剂对大鼠胰十二指肠移植后缺血-再灌注损伤的保护作用

目的：探讨大鼠胰十二指肠移植后核因子（NF）-κB活化对炎性介质表达和中性粒细胞浸润集聚的影响。方法：建立大鼠腔静脉内分泌引流、肠道外分泌引流的动物模型,脯氨酸二硫代氨基甲酸酯（ProDTC）组供体胰腺保存在ProDTC的UW液中,受体鼠于移植前30 min腹腔注射ProDTC（15 mg/kg）;对照组供体胰腺仅保存在UW液中,受体鼠腹腔注射等量生理盐水,分别于再灌注后第0、1、3、6、12、24 h等时点,处死大鼠,取血后切取移植胰腺。采用Western-blot法检测不同组胰腺组织中NF-κBp65蛋白的表达,逆转录-聚合酶链反应（RT-PCR）法检测肿瘤坏死因子-α（TNF-α）、巨噬细胞炎性蛋白-2（MIP-2）、胞间黏附分子-1（ICAM-1）mRNA表达,检测胰腺组织髓过氧化物酶（MPO）的活性,酶联免疫吸附试验（ELISA）检测血清TNF-α和MIP-2的含量。结果：与对照组相比,ProDTC组NF-κB p65蛋白和TNF-α、MIP-2、ICAM-1 mRNA表达明显下降,MPO活性明显减低（P均〈0.01）。结论：ProDTC抑制NF-κB活化,下调TNF-α、MIP-2、ICAM-1的表达,从而减轻中性粒细胞浸润及胰十二指肠移植后的缺血再灌注损伤。     

阻塞性黄疸大鼠肝脏NF-κB对炎性介质表达的调控及其在肝损伤中的作用

目的探讨阻塞性黄疸(obstructive jaund ice,OJ)时核因子κB(nuc lear factor kappa B,NF-κB)对炎性介质的调控及其在肝损伤中的作用。方法应用大鼠OJ模型,观察OJ 4、7、14 d及腹腔注射NF-κB抑制剂脯氨酸二硫化氨基甲酸酯(pyrrolid ined ith iocarbam ate,PDTC)后肝组织NF-κB P65蛋白表达、肿瘤坏死因子-α(TNF-α)mRNA、白介素-6(IL-6)mRNA、细胞间黏附分子-1(ICAM-1)mRNA表达以及血清总胆红素(total b ilirub in,TB)、谷丙转氨酶(ala-n ine transm inase,ALT)水平。结果OJ 4、7、14 d大鼠出现明显的肝损伤,表现为TB、ALT的明显增高,肝组织NF-κB P65表达明显增加,TNF-α、IL-6mRNA、ICAM-1mRNA表达也增强。注射PDTC的OJ大鼠肝NF-κB P65、ICAM-1mRNA表达较OJ各时相点显著下降,血清ALT和肝组织TNF-αmRNA较OJ在4、7 d明显下降。结论OJ时NF-κB的活化上调了炎性介质的表达,从而造成肝损伤,通过PDTC抑制NF-κB活化可减轻病程早期的肝脏损伤。     

白鹤冲剂对TNF-α刺激的血管内皮细胞与嗜中性粒细胞黏附的影响

目的:观察白鹤冲剂对人脐静脉内皮细胞(HUVEC)与人外周血嗜中性粒细胞黏附以及细胞间黏附分子-1(ICAM-1)和CD18表达及核转录因子NF-κB(NF-κB)的影响,探讨白鹤冲剂治疗血管炎的作用机制。方法:将细胞分为正常对照组、TNF-α组、TNF-α+白鹤冲剂大剂量组、TNF-α+白鹤冲剂小剂量组和TNF-α+雷公藤内酯醇组,共五组。分别进行细胞黏附实验;以细胞ELISA法观察HVVEC的ICAM-1表达;以免疫细胞化学间接法检测CD18表达;间接免疫荧光法观察白鹤冲剂对HUVEC中NF-κB活化。结果:白鹤冲剂能抑制肿瘤坏死因子-α(TNF-α)刺激的HUVEC中ICAM-1表达和人外周血嗜中性粒细胞CD18表达(P<0.05),同时呈剂量依赖性抑制人外周血嗜中性粒细胞与HUVEC的黏附。结论:通过影响HUVEC中NF-κB活化从而抑制ICAM-1表达和嗜中性粒细胞CD18的表达,最终抑制嗜中性粒细胞与HUVEC的黏附可能是白鹤冲剂治疗血管炎的作用机制。     

双参通冠方对急性心肌缺血再灌注NF-κB信号途径的影响

目的探讨双参通冠方(SSTG)对急性心肌缺血再灌注损伤心肌核因子-κB(nuclear factor-κappaB, NF-κB)信号途径(NF-κB-TNF-α-ICAM-1)的影响。方法冠状动脉前降支结扎放松法复制大鼠心肌缺血再灌注损伤模型,免疫组织化学法检测心肌组织NF-κBp65表达,酶联免疫法测定血清肿瘤坏死因子(TNF-α)、细胞间粘附分子-1(ICAM-1)含量。结果缺血再灌注模型组NF-κBp65表达较正常组明显增强,血清中TNF-α及ICAM-1含量显著升高(P<0.05),SSTG处理后NF-κBp65表达抑制,血清中TNF-α、ICAM-1水平下调(P<0.05)。结论缺血再灌注刺激可激活NF-κB信号途径,SSTG可能通过抑制NF-κB的活化,阻断NF-κB信号途径而起到心肌保护作用。     

核转录因子κB在TNF-α诱导人气道上皮细胞hBD-2表达中的作用

目的观察肿瘤坏死因子-α(TNF-α)诱导人气道上皮细胞人β防御素-2(hBD-2)的表达及核转录因子κB(NF-κB)在hBD-2表达中的作用.方法用TNF-α和或NF-κB抑制剂PDTC处理体外培养人气道原代上皮细胞,RT-PCR法检测hBD-2 mRNA的表达,Western blot检测细胞质IκB-α蛋白活性变化,凝胶迁移试验(EMSA)检测不同时相NF-κB活性的变化.结果TNF-α刺激0.5 h后可见人气道上皮细胞hBD-2 mRNA表达,并呈时间依赖性;PDTC可抑制hBD-2mRNA表达;NF-κB在TNF-α刺激1 h后明显活化,抗体超迁移率实验结果显示p65-p50异型二聚体NF-κB参与了NF-κB的活化.结论一定剂量的TNF-α可诱导人气道上皮细胞hBD-2 mRNA表达,NF-κB在TNF-α诱导气道上皮细胞hBD-2mRNA起重要的调控作用.     

还原性谷胱甘肽对急性坏死性胰腺炎肺损伤的影响

目的:观察急性坏死性胰腺炎(ANP)大鼠肺组织核因子-κB(NF-κB)mRNA,细胞因子(TNF-α,IL-1β,IL-18),髓过氧化物酶(MPO)的变化,探讨还原性谷胱甘肽(GSH)治疗肺损伤的机制. 方法:72只Wistar大鼠随机分成3组: ANP组, GSH组,SO组(假手术组). 于术后6,12,18 h取肺组织供实验用. 免疫组化测定肺中IL-1β,IL-18,TNF-α的表达,比色法测定MPO的活性,RT-PCR检测NF-κB p65 mRNA的表达,胰、肺行组织学观察. 结果:ANP组肺组织NF-κB mRNA,IL-18, IL-1β,TNF-α,MPO的水平升高,NF-κB,IL-1β,TNF-α在12 h达高峰,与SO组相比各时间点均有统计学差异(P＜0.05). GSH治疗后IL-1β,IL-18,TNF-α在各时间点均降低,NF-κB mRNA,MPO在12 h降低(P＜0.05). 结论:ANP时NF-κB,IL-18,IL-1β,TNF-α及MPO均参与了肺损伤,GSH能够抑制肺NF-κB活化,促炎因子的表达及中性粒细胞的聚集,对肺损伤有保护作用.     

核因子-κB活化与重症急性胰腺炎微循环障碍的关系

目的:探讨核因子-κB(NF-κB)活化在大白鼠重症急性胰腺炎发病中的作用与胰腺微循环障碍的关系。方法:制作大白鼠重症急性胰腺炎模型,随机分为正常组和重症急性胰腺炎组,应用免疫组化方法观察制模后1、4、12、24h胰腺组织NF-κBP65、TNF-α、IκB-α蛋白表达的动态变化;激光多普勒检测大鼠胰腺血流变化。结果:重症急性胰腺炎组胰腺组织NF-κBP65、TNF-α蛋白表达升高,IκB-α表达降低。对照组胰腺血流量无明显变化,重症急性胰腺炎组血流量明显降低。结论:NF-κB活化是重症急性胰腺炎的早期细胞内重要事件,NF-κB的活化加重了重症急性胰腺炎时胰腺的微循环障碍。     

连环蛋白p120在炎性肺损伤中保护作用的初步研究

目的探讨连环蛋白p120在炎性细胞因子肿瘤坏死因子-α(TNF-α)介导的肺微血管内皮细胞激活中的作用及其机制。方法使用小干扰RNA(siRNA)下调p120在内皮细胞中的表达,考察TNF-α刺激所引起的转录因子NF-κB激活、细胞间黏附因子ICAM-1表达水平,以及中性粒细胞跨内皮迁移能力;使用NF-κB特异性抑制剂明确p120对炎症性肺损伤的调节机制。结果 TNF-α能够降低内皮细胞p120的表达水平;p120下调加重了TNF-α介导的NF-κB激活、ICAM-1表达水平增高以及中性粒细胞跨内皮迁移,而NF-κB抑制剂则阻断了该效应。结论 p120通过抑制NF-κB活化,在TNF-α介导的肺微血管内皮细胞激活及随后的中性粒细胞跨内皮迁移中起重要的保护作用。     

黄芪提取物对大鼠全脑缺血再灌注损伤后NFκBp65、ICAM-1和TNF-α表达的影响

目的探讨黄芪提取物(EA)对大鼠全脑缺血再灌注损伤的保护机制。方法用四动脉阻断法,制备大鼠全脑缺血再灌注模型;采用Western blot检测大鼠全脑缺血再灌注6h后脑皮层组织NFκBp65和IκBα的表达;采用免疫组化(SP法)检测大鼠全脑缺血再灌注24h后脑皮层组织ICAM-1、TNF-α的表达。结果与假手术组相比,模型组大鼠全脑缺血再灌注后脑皮层组织核蛋白NFκBp65(107.1±24.18vs313.09±23.35,P<0.05)表达明显增多,而胞浆蛋白NFκBp65(221.72±19.44vs125.82±25.07,P<0.05)、IκBα(382.98±55.21vs184.37±20.40,P<0.05)表达明显减少;EA可升高大鼠全脑缺血再灌注后脑皮层组织胞浆蛋白NFκBp65、IκBα表达、降低核蛋白NFκBp65表达;同时可降低脑皮层组织ICAM-1、TNF-α的表达。结论EA的脑保护机制可能与抑制大鼠全脑缺血再灌注后NFκBp65活化及降低脑皮质中ICAM-1、TNF-α的表达有关。     

电刺激小脑顶核对局灶脑缺血再灌注大鼠脑组织核因子-κB/p65、TNF-α蛋白和基因表达的影响

目的观察电刺激小脑顶核干预对局灶性缺血再灌注大鼠脑组织核因子-κB/p65(NF-κB/p65)、肿瘤坏死因子-α(TNF-α)蛋白和基因表达的影响及其对中枢神经元保护的作用机制。方法 2014年9月—2015年9月在重庆医科大学神经病学实验室进行。Wistar大鼠60只随机分为假手术组(NC组)、缺血再灌注组(I/R组)、电刺激小脑顶核组(FNS组),每组20只,采用线栓法制作脑缺血再灌注动物模型,FNS组予以电刺激小脑顶核干预,造模或干预成功后处死大鼠取脑组织,分别采用免疫组织化学法和RT-PCR检测各组NF-κB/p65、TNF-α的表达。结果与NC组比较,I/R组NF-κB/p65、TNF-α蛋白表达增加(q=0.032、0.020,P<0.05);与I/R组比较,FNS组NF-κB/p65、TNF-α的表达明显降低(q=0.182、0.013,P<0.05)。与NC组比较,L/R组NF-κB/p65mRNA和TNF-αmRNA表达量明显增高(P<0.05);与I/R组比较,FNS组NF-κB/p65 mRNA和TNF-αmRNA表达量明显减少(P均<0.05)。结论电刺激小脑顶核抑制NF-κB/p65的活化和TNF-α的转录,NF-κB/p65的活化抑制和表达减少及所致的TNF-α表达下调可能是其发挥中枢神经源性保护作用的机制之一。     

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.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

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.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.