黄芩素对D-氨基半乳糖联合脂多糖所诱导的小鼠肝损伤的保护作用

[目的]建立D-氨基半乳糖联合脂多糖(D-GalN/LPS)所诱导的小鼠肝损伤模型,探讨黄芩素对肝损伤的保护作用.[方法]腹腔注射给予小鼠D-GalN/LPS,分别于给药1,2,4,6h后采血并剖取肝脏,测定血清ALT,AST,TNF-α值及肝组织GSH水平.另取小鼠适量制作肝损伤模型,分别灌胃给予黄芩素50,100,150mg/kg,1h后测定血清ALT,AST,TNF-α值.[结果]给予D-GalN/LPS6h时小鼠血清ALT,AST水平上升最显著,2h时血清TNF-α水平上升最显著,4h时开始下降,6h时趋于正常,小鼠肝匀浆中GSH水平在6h时损耗最显著.各剂量黄芩素均可降低D-GalN/LPS诱导的肝损伤小鼠的血清ALT,AST,TNF-α水平.[结论]黄芩素对D-GalN/LPS所诱导的小鼠急性肝损伤具有保护作用.     

LPS/D-GalN诱发NF-κB转基因小鼠急性致死性肝损伤模型的建立

目的 以NF-κB转基因Balb/C小鼠为实验动物,建立一个LPS/D-GalN诱发的急性致死性肝损伤模型,既一个极端的重症肝损伤模型。方法 采取腹腔注射高剂量的LPS/D-GalN建立急性致死性肝损伤小鼠模型,观察模型小鼠的促炎症细胞因子水平和NF-κB的活性改变,以及肝脏功能和病理改变情况。结果 模型组小鼠生存时间为8-10h,模型建立后小鼠血清TNF-?、IL-6和MCP-1水平显著升高,在2-4小时达到高峰;肝脏外观出现瘀血和出血,肝脏小叶被严重破坏,肝细胞严重坏死和出血;血清ALT/AST水平在模型诱发后持续迅速上升;整体成像显示NF-κB的活性在4-6h达到高峰。正常对照组小鼠以上指标无显著变化。结论 成功建立LPS/D-GalN诱发的NF-κB转基因小鼠的急性致死性肝损伤模型。     

新型造影剂在小鼠肝脏肿瘤Micro-CT活体成像中的应用

目的 利用新型纳米颗粒造影剂结合Micro-CT成像技术,建立小鼠肝脏成像方法,并用于肝脏肿瘤的活体成像.方法 6只6～8周龄雄性C57BL/6J小鼠随机分成A组和B组,分别尾静脉注射纳米颗粒造影剂ExiTron nano 12000 50 μL和100 μL;在注射前、注射后3 min、24 h、7 d、14 d、28 d和56 d对所有小鼠肝脏进行Micro-CT活体扫描;分别在小鼠肝左叶和肝右叶内选取感兴趣区(ROI)进行灰度值分析,比较不同时间点肝组织对比度的变化.确定合适的造影剂剂量,尾静脉注射至3只雄性16月龄HBV转基因肝癌模型小鼠(C组),同上进行Micro-CT活体扫描,并于第56天全部安乐死后取肝脏观察病理学改变.结果 A组和B组小鼠在注射不同浓度造影剂后,冠状位重建图像及肝脏感兴趣区的平均灰度值结果显示:肝脏实质造影后均比注射前明显增强,24 h达到峰值,注射后56 d内,小鼠肝脏感兴趣区的平均灰度值与注射前相比仍维持在较高的水平,B组显著高于A组(P<0.01),确定后续实验采用B组造影剂剂量(100 μL).C组注射100 μL造影剂后,各时间点均能比较清楚地看到肝脏癌性结节存在,病理学观察发现肝脏出现非典型增生,肿瘤细胞核大,染色质加深和肝细胞坏死.结论 利用纳米颗粒造影剂结合Micro-CT成像技术,成功建立了小鼠肝脏活体成像方法,并可应用于肝脏肿瘤的活体成像研究.     

外源性低浓度一氧化碳对小鼠急性肝损伤的保护作用研究

目的 研究外源性低浓度一氧化碳(CO)对D-氨基半乳糖(GAlN)和脂多糖(LPS)联合诱导小鼠急性肝损伤的保护作用及机制.方法 将野生型C57BL/6小鼠分为正常对照组、CO对照组、肝损伤组和肝损伤+CO组(GalN/LPS+CO组),每组8只.肝损伤组小鼠经腹腔注射GalN(550 mg/kg)和LPS(15 mg/kg)造模.CO对照组:经腹腔注射纯品CO气体(15 ml/kg),6h后按8ml/kg剂量再注射一次,期间动态检测碳氧血红蛋白(HbCO)的水平变化.GalN/LPS+CO组:按同样方法注射CO气体,12h后再给予GalN/LPS染毒.染毒12h后检测各组小鼠的肝损伤指标、病理学指标、细胞凋亡和炎症因子(IL-1b、IL-6、IL-10和TNF-a)的水平变化.结果 小鼠给予GalN/LPS后发生严重急性肝损伤;与正常对照组或CO对照组相比,染毒组小鼠血浆ALT、AST和T-BIL水平均明显升高,大量肝细胞变性、坏死.小鼠腹腔注射低浓度CO使血HbCO的水平维持在8％～12％达12h以上,显著降低GalN/LPS染毒导致的ALT、AST和TBIL水平升高,同时明显减轻肝细胞凋亡和抑制炎症因子水平.结论 外源性低浓度CO可以减轻GalN/LPS诱导的小鼠急性肝损伤,其机制可能与降低细胞凋亡和细胞因子的水平有关.     

地西泮和莫达非尼对小鼠试验性急性肝衰竭的影响

目的：研究地西泮、莫达非尼对小鼠试验性急性肝衰竭的影响。方法：雄性昆明小鼠240只,给予D-半乳糖胺(D-GalN)和脂多糖（LPS）同时腹腔注射,制备小鼠急性肝衰竭模型;并于造模前2h,治疗组分别给予相应药物灌胃,对照组给予蒸馏水灌胃。比较各组小鼠的24h存活率,血清转氨酶水平,肝脏组织病理学改变,血清TNF-α,IL-1的水平,肝脏组织中SOD,MDA ,GR,１GSH,NO,NOS的水平。结果：地西泮可以提高小鼠存活率,减轻肝脏病变程度,降低血清转氨酶活性及TNF-α,IL-1的浓度,增加肝脏组织中SOD,GR的活性,降低NOS的活性及MDA,NO的浓度。莫达非尼可加重肝脏病变程度,升高血清转氨酶的活性及 TNF-α,IL-1的浓度,降低肝脏组织中SOD,GR的活性及升高MDA的浓度。结论 ：地西泮可减轻D-GalN/LPS所致的急性肝衰竭,提高小鼠的存活率,莫达非尼可加重小鼠的肝损伤。     

N-乙酰半胱氨酸对GalN/LPS引起小鼠急性肝损伤的保护作用

目的研究N-乙酰半胱氨酸(NAC)对D-氨基半乳糖(D-GalN)与细菌脂多糖(LPS)引起小鼠急性肝损伤的保护作用。方法建立GalN/LPS引起的小鼠急性肝损伤模型。在GalN与LPS共处理前30min经腹腔注射给予NAC,于GalN/LPS处理1·5h后剖杀部分动物取血,测定血清肿瘤坏死因子α(TNF-α)含量;GalN/LPS处理8h后剖杀剩余动物,取血和肝脏,测定血清丙氨酸氨基转移酶(ALT)活力和一氧化氮(NO)水平,检测肝脏组织Caspase-3活性与还原性谷胱甘肽(GSH)含量,采用DNA断裂分析方法检测肝脏凋亡,并对部分肝脏进行病理组织学检查。结果与GalN/LPS处理组比较,NAC预处理组小鼠血清ALT活力、肝脏Caspase-3活性和GSH损耗均显著下降,肝脏组织病理学改变明显改善,DNA断裂分析未见明显凋亡梯度,而NAC对小鼠血清TNF-α含量和NO生成无显著影响。结论NAC预处理对GalN/LPS引起的小鼠急性肝损伤有保护作用,主要通过其抗凋亡和抗氧化作用。     

灵芝酸A对D-氨基半乳糖/脂多糖诱导小鼠肝损伤的保护作用

目的?研究灵芝酸A对D-氨基半乳糖(D-GaIN)/脂多糖(LPS)诱导小鼠肝损伤的保护作用及其相关机制。方法?60只C57BL/6小鼠随机分为4组:正常组、模型组、灵芝酸A低剂量组（20?mg/kg）、灵芝酸A高剂量组（40?mg/kg）。除正常组和模型组给予等体积蒸馏水外,其余各组每天灌胃给予相应剂量药物,共7?d。末次给药4?h后,腹腔注射D-GalN(700?mg/kg)与LPS(1?mg/kg),正常组腹腔注射等体积溶媒。24?h后取血、肝脏。ELISA法检测血清谷草转氨酶（AST）、谷丙转氨酶（ALT）活性以及白介素（IL-6、IL-1β）和肿瘤坏死因子-α（TNF-α）含量,各组小鼠肝脏做HE染色,并采用蛋白免疫印迹法（Western blot）检测肝脏组织中NLRP3/NF-κB蛋白表达。结果?灵芝酸A（20、40?mg/kg）能显著降低D-GaIN/LPS诱导的肝损伤小鼠血清ALT、AST活性及IL-6、IL-1β和TNF-α含量;能显著改善小鼠肝组织的病理学改变;显著降低肝脏NLRP3/NF-κB蛋白表达。结论?灵芝酸A对D-GaIN/LPS诱导的小鼠肝损伤具有保护作用,这种效应可能与调控NLRP3/NF-κB信号通路有关。      

急性肝损伤小鼠肝组织中的自噬现象

目的探讨在脂多糖(lipopolysaccharide,LPS)和D-氨基半乳糖(D-galactosamine,D-GalN)诱导的小鼠急性肝损伤模型中,肝脏组织的自噬情况。方法将60只雄性Balb/C小鼠随机分为5组(每组均包括实验亚组和对照亚组,各6只)。实验组小鼠腹腔注射LPS(100μg/kg)和D-GalN(350mg/kg)溶液,对照组小鼠只注射无菌等量生理盐水,分别于注射后2、4、8、12、24h处死小鼠,取其血液及肝脏。另取3只小鼠饥饿处理24h作为自噬阳性对照组。所有小鼠于处死前24h经高压尾静脉注射法注射溶于2ml生理盐水的GFP-LC3质粒100μg。生化法检测血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)水平;透射电镜、荧光显微镜下观察小鼠肝脏的自噬情况。结果各实验组血清ALT、AST水平均明显高于相应对照组(P<0.01)。透射电镜显示,实验组和自噬阳性对照组肝组织中可见大量肝细胞发生自噬。荧光显微镜下可见GFP-LC3绿色荧光蛋白。8、12、24h实验组及自噬阳性对照组每高倍镜视野下平均GFP-LC3荧光点数目明显高于对照组(P<0.01)。结论 LPS和D-GalN可诱导小鼠发生急性肝损伤,而自噬是这一过程中的重要形态学特征。     

脂多糖预处理减轻对乙酰氨基酚所致急性肝损伤

目的：建立对乙酰氨基酚( APAP)引起小鼠急性肝损伤动物模型,探讨低剂量脂多糖( LPS)预处理对APAP引起急性肝损伤的作用。方法①选取ICR雄性小鼠20只,随机分成APAP组和LPS+APAP组。禁食后,APAP组经腹腔注射APAP(300 mg/kg);LPS+APAP组经腹腔注射先给予LPS(50μg/kg),3 h后再给予APAP(300 mg/kg),密切观察小鼠存活状况,72 h后取材,测生化指标,行肝组织 HE 染色。②ICR雄性小鼠随机分成APAP 0、0．5、6、12、24、72 h组和LPS+APAP 0、0．5、6、12、24、72 h组。禁食后,APAP组腹腔注射APAP(300 mg/kg);LPS+APAP组腹腔注射LPS(50μg/kg),3 h后再经腹腔注射APAP(300 mg/kg),分别在每组对应时点取材。分离血清测丙氨酸氨基转移酶( ALT),用Beutler改良法测肝脏谷胱甘肽( GSH )含量。结果①APAP组给药4 h左右小鼠开始出现死亡情况,72 h存活率为50%;LPS+APAP组72 h内无死亡。②低剂量LPS预处理能降低血清ALT(P<0．05),减少小鼠肝脏坏死情况(P<0．01)。③ APAP组与LPS+APAP组肝脏GSH含量差异无统计学意义。结论低剂量LPS预处理能显著减轻APAP引起的急性肝损伤,这种作用不是通过减弱N-乙酰苯醌亚胺( NAPQI)耗竭GSH而产生的,可能与其激活了机体的免疫应答,抑制了GSH耗竭的下游机制有关。     

LPS信号转导分子TLR4表达与小鼠肝损伤的关系

目的 :探讨脂多糖 (lipopolysaccharide ,LPS)信号转导分子TLR4(Toll likereceptor 4 ,TLR4)与肝脏损伤的关系。方法 :小鼠腹腔注射LPS后不同时间点取肝脏组织 ,HE染色观察病理改变 ,RT PCR法检测肝组织TLR4mRNA的表达。结果 :腹腔注射LPS后 2 4h小鼠肝脏出现明显炎性浸润、坏死 ,肝组织TLR4mRNA的表达在注射LPS后 6 ,1 2h明显抑制 ,2 4h则基本恢复。结论 :LPS致肝损伤呈时间依从性 ;LPS信号转导分子TLR4在介导LPS肝损伤中起重要作用     

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.