降钙素基因相关肽对大鼠肝脏缺血-再灌注损伤的保护作用

本实验观察到,大鼠肝脏缺血-再灌注引起血清谷丙转氨酶(GPT)和谷草转氨酶(GOT)活性明显升高,若预先给予降钙素基因相关肽(CGRP)则使GPT和GOT的漏出明显减轻。但在离体培养的大鼠肝细胞,CGRP对自由基生成系统黄嘌呤-黄嘌呤氧化酶引起的细胞损伤未见有直接的保护作用。     

右旋儿茶素对四氯化碳或半乳糖胺引起原代培养大鼠肝细胞毒性的作用

利用原代培养大鼠肝细胞的方法,研究右旋儿茶素(d-CTC)对四氯化碳(CCl_4)或半乳糖胺(d-GalN)引起肝细胞毒性的作用,并观察该药对肝酶的直接作用。结果表明:d-CTC 0.6～5.0mg/ml可使含10mmol/L CCl_4或5mmol/Ld-GalN肝细胞培养液中的GOT.GPT及LDH活性显著降低,并呈量效关系。d-CTC 2.5～5.0mg/ml可直接抑制LDH及GPT活性,但对GOT无明显影响。其抑酶作用与抗肝毒而降低酶活性比较,有非常显著的差别,提示d-CTC具有较强的抗肝毒作用。     

氧化苦参碱对四氯化碳损伤大鼠肝细胞的保护作用

目的 :研究氧化苦参碱对肝细胞的保护作用。方法 :采用原代肝细胞培养的方法 ,选用CCl430mmol/L制备肝损伤模型 ,同时加以不同浓度的氧化苦参碱以保护肝细胞 ,培养 12h后以MTT法测细胞存活率 ,并测定培养基中AST、ALT、LDH。结果 :氧化苦参碱对肝细胞的保护呈剂量依赖性 ,氧化苦参碱组细胞存活率明显高于CCl4损伤组 (98.3% /3.2 % ,P <0 .0 5 ) ;其AST、ALT、LDH较CCl4损伤组明显降低 (依次为 6 .13± 0 .4 4 /896 .87±13.2 3、2 1.5± 0 .76 /15 0 2 .13± 35 .80、19.5± 0 .6 8/2 87.38± 5 .79,均P <0 .0 5 )。结论 :氧化苦参碱对CCl4损伤的肝细胞具较好的保护作用 ,存在最佳剂量性关系。     

几种胃肠肽对四氯化碳所致大鼠急性肝损伤的保护作用及其机制

目的　研究表皮生长因子 (EGF)、神经降压素 (NT)、降钙素基因相关肽 (CGRP)对四氯化碳 (CCl4)急性损伤的在体大鼠肝脏和离体培养肝细胞的保护作用 ,并探讨其机制。　方法　第一步建立大鼠 CCl4损伤模型 ,设立对照组、损伤组及各胃肠肽保护组 ,于注射 CCl4前 30 min、前 10 min,后 10 min注射各胃肠肽 ,2 4h后测定血清酶学水平、肝匀浆 SOD活性及 MDA含量 ,观察肝脏形态学变化 ;第二步建立大鼠离体培养肝细胞损伤模型 ,设对照组、损伤组及不同浓度胃肠肽预处理组 (胃肠肽提前 1h加入 ) ,2 4h后测上清液酶学水平、肝细胞内 SOD活性和 MDA含量 ,以台盼蓝染色试验计算肝细胞存活率 ,观察形态学变化 ;第三步以 Fura- 2 / AM和 DPH荧光探针测定肝细胞内游离钙和膜流动性 ,观察对肝细胞有直接保护作用的胃肠肽对这两个指标的影响。　结果　 (1) EGF、NT、CGRP明显降低 CCl4损伤后大鼠血清酶学水平、肝匀浆 MDA含量 ,使 SOD活性回升 (P<0 .0 1) ,改善肝脏病理学变化。(2 ) EGF、NT明显降低原代培养肝细胞 CCl4损伤后上清液酶学水平、MDA含量 ,提高 SOD活性和细胞存活率 (P<0 .0 1) ,改善形态学变化。CGRP对上述指标和形态学改变无明显影响。(3) EGF、NT明显对抗肝细胞 CCl4损伤后胞内 [Ca2 + ]i的增高和膜流动     

病毒性肝炎肝细胞损伤程度对血清转氨酶活性与比值的影响

本文对组织学和血清学检查确诊为病毒性肝炎的98例患者分析了肝细胞损伤程度对血清转氨酶活性及GOT/GPT比值的影响。结果表明①转氨酶活性与肝细胞损伤程度基本一致,但并不完全平行。②GOT/GPT比值与肝损伤程度关系密切。提示:仅靠转氨酶高低诊断肝炎有一定的局限性,观察GOT/GPT比值对判断肝损伤程度有重要意义。     

微囊化异种肝细胞移植对药物性肝衰的治疗作用及免疫隔离机制的初步研究

目的 :探讨微囊化异种肝细胞脾内移植对药物性肝衰大鼠的治疗作用 ;测定受体存活率、肝功能的变化及CD4 ,CD8的变化。方法 :海藻酸钠体外包裹经胶原酶技术制备的异种 (豚鼠 )肝细胞为供体 ,SD大鼠为受体 ,D -氨基半乳糖 (19.5ml/kg)腹腔内一次性注射 ,制作肝衰模型。 4 8h后将微囊化的游离豚鼠肝细胞 (1.5× 10 7)移植于大鼠脾脏内。以豚鼠裸肝细胞 (1.5× 10 7)移植及生理盐水1.2ml脾脏注射为对照。移植后 14d观察存活率、肝功能及肝脏病理情况 ,免疫组化ABC法测定CD4 ,CD8的变化。结果 :微囊化肝细胞移植组存活率 80 % ,明显高于生理盐水组 (2 5 % )和裸肝细胞移植组(70 % )。 (P <0 .0 1　V　P <0 .0 5 )。微囊肝细胞移植组 14d总胆红素 (7.99± 0 .5 8mg/L)和ALT(5 5±6 .7u) ,明显低于生理盐水组 (9.6 3± 0 .83mg/LV 91± 8.0u)和裸肝细胞组 (8.9± 0 .6 6mg/LV 74± 7.1u)(P <0 .0 5VP <0 .0 1)。移植后 12h ,72h ,14d分别测定受体脾脏CD4 ,CD8淋巴细胞 ,微囊化肝细胞组72h ,14d呈阳性。裸肝细胞组均呈阳性。结论 :大鼠药物性肝衰微囊化肝细胞脾内移植后维持存活率14d ,细胞免疫参与排斥反应 ,肝细胞微囊化处理可延迟细胞免疫的发生时间。     

重症急性胰腺炎急性肾损伤机制探讨

目的：通过体内体外观察大鼠和大鼠正常肾细胞(NRK)在重症急性胰腺炎（severe acute pancreatitis, SAP）状态下凋亡和分泌肿瘤坏死因子α(TNF-α)的情况,探讨SAP时急性肾功能损伤的机制。方法: SD大鼠54只随机分成SAP组（n=30）和假手术组(n=24),SAP组以5%牛磺酸胆酸钠溶液胆管逆行注射诱导模型,术后6、12、24h采集血清、腹水。ELISA法检测血清、腹水TNF-α水平,TUNEL法检测肾组织凋亡率,RT-PCR检测肾组织TNF-αmRNA表达。将24h采集的血清和腹水处理后,体外培养NRK细胞,MTT法检测细胞活性,检测培养液TNF-α水平,流式细胞术测定NRK细胞凋亡率,检测NRK细胞TNF-αmRNA表达。 结果: SAP组6、12、24h血清和腹水TNF-α水平呈进行性升高,各时段比较差异有统计学意义(P<0.05),较假手术组在各时段明显升高(P<0.01)。SAP组肾组织TNF-α　mRNA表达较假手术组明显上调。SAP组凋亡指数在6、12、24h时分别为（18.6±5.6）%、（18.1±4.7）%、（25.3±5.1）%,假手术组分别为（19.4±3.1）%、（13.5±2.8）%、（14.6±5.5）%, 24?h时SAP组凋亡指数明显高于假手术组（P<0.05）。SAP组24h肾组织TNF-α mRNA的表达较假手术组明显上调。经SAP组大鼠血清和腹水处理后,NRK细胞活性低于假手术组。经SAP组大鼠血清和腹水处理24h后,NRK细胞培养液中TNF-α水平较假手术组明显升高（P<0.01）,NRK细胞TNF-αmRNA表达上调,显著高于假手术组,NRK细胞凋亡率均较假手术组明显升高（P<0.05）。结论: SAP时肾损害明显,其损伤可能与肾细胞过度凋亡有关,而肾细胞过度凋亡与血清和腹水TNF-α升高及肾细胞过多分泌TNF-α有关,SAP腹水对肾的直接损伤可能比血清明显。     

大鼠肝脏缺血─再灌注损伤时肝细胞线粒体膜电位的变化

目的:探讨肝脏缺血—再灌注损伤的机制。方法:将健康Wistar大鼠30只,随机分成二组:假手术组(Controlgroup),缺血—再灌注组(I-R group)。假手术组结扎肝镰状韧带,钝性游离左、中叶肝蒂,不作结扎。缺血—再灌注组,用无损伤动脉夹夹闭左、中叶肝蒂,缺血1h,再灌注1h,制成部分(70%)缺血—再灌注模型。用荧光染料罗丹明(Rh123)和碘化丙啶(PI)标记肝细胞,流式细胞仪(FCM)测定线粒体膜电位(ΔΨm),表示为Rh123+PI-细胞和Rh123-PI-细胞所占的百分数。结果:缺血—再灌注后,假手术组大鼠Rh123+PI-细胞占(94.2±2.8)%,Rh123-PI-细胞占(3.9±0.9)%。缺血—再灌注组肝细胞ΔΨm降低,Rh123-PI-细胞(14.1±2.1)%,与假手术组相比明显增多(P<0.01),Rh123+PI-细胞占(83.3±3.9)%,与假手术组相比明显减少(P<0.01)。结论:线粒体膜电位的变化介导了肝脏缺血—再灌注损伤的机制。     

大鼠肝脏缺血-再灌注损伤时肝细胞线粒体膜电位的变化

目的探讨肝脏缺血-再灌注损伤的机制.方法将健康Wistar大鼠30只,随机分成二组假手术组(Control group),缺血-再灌注组(I-R group).假手术组结扎肝镰状韧带,钝性游离左、中叶肝蒂,不作结扎.缺血-再灌注组,用无损伤动脉夹夹闭左、中叶肝蒂,缺血1h,再灌注1h,制成部分(70%)缺血-再灌注模型.用荧光染料罗丹明(Rh123)和碘化丙啶(PI)标记肝细胞,流式细胞仪(FCM)测定线粒体膜电位(ΔΨm),表示为Rh123+PI-细胞和Rh123-PI-细胞所占的百分数.结果 缺血-再灌注后,假手术组大鼠Rh123+PI-细胞占(94.2±2.8)%,Rh123-PI-细胞占(3.9±0.9)%.缺血-再灌注组肝细胞ΔΨm降低,Rh123-PI-细胞(14.1±2.1)%,与假手术组相比明显增多(P<0.01),Rh123+PI-细胞占(83.3±3.9)%,与假手术组相比明显减少(P<0.01).结论线粒体膜电位的变化介导了肝脏缺血-再灌注损伤的机制.     

大豆磷脂对四氯化碳致肝细胞损伤的保护作用

目的 观察大豆磷脂脂质体(SPL)的累积对四氯化碳导致肝细胞损伤的保护作用.方法 新生的小鼠肝细胞原代培养24 h后分为正常对照组、损伤组和SPL保护组.损伤组采用CC14作用于培养的小鼠肝细胞3 h:SPL保护组在加入损伤因素的同时分别加入不同浓度(0.2、0.4、0.8和1.6 mmol/mL)的SPL继续培养3 h.观察细胞生长情况.采用MTT法测定肝细胞的存活率;测定肝细胞培养液中乳酸脱氢酶(LDH)和谷丙转氨酶(ALT)的活力,以及肝细胞丙二醛(MDA)的含量和超氧化物歧化酶(SOD)的活力.结果 与正常对照组比较,损伤组肝细胞存活率明显降低(P＜0.01),培养液中LDH、ALT释放量明显增加(P＜0.01),肝细胞中SOD活力显著降低(P＜0.01),MDA含量显著升高(P＜0.01);SPL保护各组与损伤组相比,细胞存活率均明显升高(P＜0.01),培养液中LDH、ALT释放明显减少(P＜0.01),肝细胞中MDA含量显著降低(P＜0.01),SOD活力显著升高(P＜0.01),并均呈一定的剂量依赖关系.结论 大豆磷脂脂质体对四氯化碳致肝细胞损伤具有明显的保护作用.     

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.