BABL/c小鼠感染大肠杆菌O127模型的建立及TGF-β1因子的荧光定量检测

目的 建立BABL/C小鼠感染大肠杆菌O127的模型，并建立TGF-β1的荧光定量PCR方法检测方法。方法 通过灌胃的方式建立肠炎性小鼠模型，并利用嵌合荧光定量的方法对其结肠TGF-β1定量分析实验。结果 成功建立了肠炎性小鼠模型，荧光定量PCR试验结果表明实验组TGF-β1表达量明显高于对照组。结论 利用经口灌胃的方法可以成功的建立肠炎性小鼠动物模型，TGF-β1定量方法可以作为模型建立的评价标准。     

P300介导VEGF下调致孕期酒精暴露模型子代小鼠宫内生长受限

目的 探讨P300介导血管内皮生长因子(VEGF)下调致孕期酒精暴露模型子代小鼠宫内生长受限的可能机制。方法 选取SPF级c57小鼠60只。按照雌雄2∶1合笼,次日清晨观察雌鼠阴栓,有阴栓者即为妊娠成功(妊娠成功30只),记妊娠0.5 d,即胎鼠胎龄E 0.5 d。此时开始给予酒精灌胃(5μL/g/d 50%乙醇)建立胎儿酒精综合征(FAS)模型设为实验组,另设对照组(予生理盐水灌胃),实验组及对照组每组各12只[6只雌鼠因灌胃失败未纳入实验]。检测出生体质量;利用化学比色法测定胎盘组织中组蛋白乙酰化酶活性;利用荧光测定法测定胎盘组织中组蛋白去乙酰化酶活性;定量PCR检测组蛋白乙酰化酶p300和PCAF表达,定量PCR和Western blot检测VEGF表达;ChIP-PCR技术检测p300与VEGF启动子结合水平。结果 与对照组相比,实验组小鼠出生体质量显著降低(P<0.01),且存在出生后生长发育迟缓;实验组小鼠胎盘中组蛋白乙酰化酶活性较对照组明显下降(P<0.05),而去乙酰化酶活性两组比较差异无统计学意义(P>0.05);实验组小鼠胎盘组织中p300、VEG...     

岩藻多糖对炎症性肠病的作用及其机制

目的：探讨岩藻多糖抑制炎症性肠病（inflammatory bowel disease,IBD）的作用及其机制。方法：将小鼠随机分为实验组和对照组（每组5只）,实验组给予岩藻多糖灌胃2周,对照组给予饮用水灌胃2周,随后两组小鼠均自由饮用溶有葡聚糖硫酸钠（dextran sulfate sodium salt,DSS）的水溶液,构建IBD小鼠模型。检测小鼠体重、脾脏重量、结肠长度及重量,HE染色观察结肠组织病理变化,观察大便性状并评分;运用实时荧光定量PCR（real?time fluorescence quantitative PCR,qPCR）检测炎症因子、趋化因子和紧密连接蛋白的mRNA水平变化。结果：岩藻多糖处理的实验组小鼠肠炎相关指标显著优于对照组,炎症程度减弱。结论：岩藻多糖能够预防和抑制IBD的发生,为预防和治疗IBD提供新方法和依据。     

骆驼奶对噁唑酮诱导的小鼠肠炎的作用

目的 探讨骆驼奶对噁唑酮诱导的小鼠肠炎模型中肠道黏膜免疫功能的调节作用.方法 将40只BALB/c雄性小鼠以3％噁唑酮乙醇溶液涂搽皮肤致敏后用1％噁唑酮乙醇溶液灌肠,建立小鼠肠炎模型,分为实验组和对照组,每组20只,每天分别用骆驼奶和双蒸水灌胃.记录小鼠体质量、疾病活动指数;采用流式细胞术、ELISA、实时荧光定量PC...     

IL-27在小鼠实验性肠炎模型中的表达

目的:通过检测IL-27在小鼠实验性肠炎模型中的表达,探讨其在肠炎发病机制中的作用。方法:建立三硝基苯磺酸(TNBS)诱导的急性小鼠实验性肠炎模型,采用实时荧光定量PCR(qRT-PCR)及Western blot分别检测IL-27基因mRNA和蛋白在炎症肠黏膜中的表达。结果:TNBS成功诱导急性小鼠实验性肠炎,与对照组相比,TNBS处理的小鼠炎症评分明显增高,炎症肠黏膜中IL-27mRNA和蛋白表达亦明显增高,差异具有统计学意义。结论:IL-27在TNBS诱导的小鼠炎症肠黏膜中表达明显增高,提示IL-27可能参与小鼠肠道炎症的形成过程。     

鼠李糖乳杆菌效应蛋白HM0539增强小鼠对大肠杆菌O157:H7肠道感染的抵抗力

目的 探究鼠李糖乳杆菌LGG的效应蛋白HM0539对大肠杆菌O157∶H7肠道感染的预防效果。方法 采用黏附、侵袭实验评价HM0539处理后,大肠杆菌O157∶H7对HT-29细胞的黏附侵袭能力。通过免疫荧光、免疫印迹等方法进一步检测在使用HM0539治疗过程中对HT-29细胞中黏蛋白（MUC2）、紧密连接蛋白（ZO-1）表达的影响;通过动物实验,观察小鼠的生存率及体质量变化,检测攻毒小鼠的肠道病理及肠道屏障功能的改变。结果 HM0539呈浓度依赖性地抑制大肠杆菌O157∶H7黏附和侵袭HT-29,且HM0539的预处理效果优于共同处理（P<0.05）;免疫荧光和免疫印迹等结果显示,HM0539不仅可以明显降低大肠杆菌O157∶H7感染后MUC2的表达水平（P<0.05）,还可显著抑制其对细胞间紧密连接蛋白的破坏（P<0.05）。动物实验结果证 明HM0539可抑制攻毒小鼠的体质量下降（P<0.05）和空肠的损伤;HM0539可抑制大肠杆菌O157∶H7诱导的空肠杯状细胞的破坏（P<0.05）;此外,HM0539可上调攻毒小鼠空肠中的MUC2、ZO-1蛋白的表达（P<0.05）。结论 HM0539可抑制大肠杆菌O157∶H7黏附和侵袭HT-29细胞,而且增强了小鼠对大肠杆菌O157∶H7感染的抵抗力,其机制可能是通过抑制大肠杆菌O157∶ H7破坏黏蛋白和细胞间紧密连接蛋白。     

实时荧光定量PCR法分析苦参对大鼠胃肠道大肠杆菌数量的影响

目的:应用实时荧光定量PCR技术分析口服苦参对大鼠胃、十二指肠、空肠、回肠、结肠部位以及粪便内大肠杆菌数量的影响,探索苦参对胃肠道中大肠杆菌菌群调节作用。方法:常规饲养大鼠150只,苦参水煎液分高、中、低剂量灌胃,另设空白对照组与抗生素组各10只。分别在第一、二、三周时采集大鼠胃、十二指肠、空肠、回肠、结肠各段(含内容物)以及粪便,提取细菌基因组DNA,采用实时荧光定量PCR法(绝对定量方法)检测各样本不同给药时间点大肠杆菌的数量。结果:(1)与空白对照组相比较:各给药组十二指肠部位在第一、二、三周时,大肠杆菌数量显著降低(P0.01);在不同剂量组不同给药时间点,胃部、空肠、回肠部位以及粪便中大肠杆菌数量均有不同程度降低(P0.05)。(2)与抗生素组比较:低、中、高剂量组给药二周后,十二指肠、空肠、回肠部位大肠杆菌数量明显偏低(P0.05),而结肠及粪便部位在给药一周后高剂量组大肠杆菌数量明显偏高(P0.01)。结论:苦参水煎液灌胃可造成大鼠十二指肠、空肠、回肠部位大肠杆菌菌群数量降低,其中对十二指肠部位大肠杆菌数量的影响随时间推移呈递减趋势,与抗生素的作用基本一致。     

不同配比与剂量黄芪当归对药对肺纤维化小鼠肺形态学及羟脯氨酸含量影响

目的:观察黄芪、当归不同剂量与配伍比例对实验性肺纤维化小鼠肺病理形态学及肺组织羟脯氨酸(HYP)含量的影响。方法:SPF级ICR雄性小鼠80只,体重18~22 g,随机分为正常对照组、模型组,以及6个不同配比与剂量的黄芪当归组成的中药治疗组(1.大剂量黄芪当归5∶1组、2.大剂量黄芪当归1∶1组、3.大剂量黄芪当归1∶5组、4.小剂量黄芪当归5∶1组、5.小剂量黄芪当归1∶1组、6.小剂量黄芪当归1∶5组),每组10只,除正常对照组外,其余各组采用气管内注射博莱霉素(5 mg/kg)复制小鼠肺纤维化模型。造模第2天起,正常对照组和模型组用生理盐水灌胃,6个中药治疗组给予不同配比与剂量的黄芪当归水煎液灌胃治疗,于28 d处死小鼠,观察肺组织病理变化及羟脯氨酸的含量。结果:HE染色显示模型组小鼠支气管周围及肺泡间隔有大量炎症细胞浸润,肺泡壁增厚,肺泡结构破坏甚至消失,肺泡间隔纤维化显著。武兆发简化Mallory氏胶原染色显示模型组小支气管壁周围有大量蓝色胶原纤维沉积。6个中药治疗组较模型组均有所改善,其中中药治疗第1、6组改善明显,肺泡结构已基本恢复。与模型组比较,中药第1、6组小鼠肺组织Hyp含明显降低,统计学差异显著(P<0.01)。结论:大剂量黄芪当归5∶1组在降低肺组织Hyp含量及改善肺泡结构方面效果显著。     

淫羊藿苷对前列腺癌原位移植瘤小鼠肿瘤生长及细胞周期的影响

目的 探讨淫羊藿苷对前列腺癌原位移植瘤SCID小鼠肿瘤生长、细胞周期、磷酸酶及张力蛋白同源物(phosphatase and tensin homolog,PTEN)基因表达的影响。方法 40只雄性SCID小鼠,根据随机数字表法随机均分为模型对照组、低剂量组、中剂量组、高剂量组,每组10只。采用SCID小鼠前列腺腺体背外侧包膜内注射人前列腺癌LNCaP细胞株悬液的方法构建前列腺癌原位移植瘤小鼠模型。造模2周后,模型对照组予以0.9%氯化钠注射液,低剂量组予以10mg/kg淫羊藿苷,中剂量组予以40mg/kg淫羊藿苷,高剂量组予以80mg/kg淫羊藿苷。灌胃处理1次/d,治疗时间为5周。在灌胃给药治疗前及治疗5周后,分别取各组小鼠 5只,对比各组肿瘤质量、肿瘤体积、肿瘤抑制率。采用流式细胞学方法检测LNCaP细胞周期,计算各周期比值。采用实时定量PCR检测前列腺肿瘤组织中PTEN mRNA相对含量。结果 治疗后,模型对照组肿瘤质量、肿瘤体积较治疗前明显增加(P<0.05);中剂量组及高剂量组肿瘤质量、肿瘤体积较治疗前明显降低(P相似文献   

牛初乳通过调节肠道菌群对小鼠溃疡性结肠炎的改善作用

为探讨牛初乳(BC)对小鼠溃疡性结肠炎的改善作用,采用葡聚糖硫酸钠(DSS)构建肠炎模型,监测小鼠体重、肠炎评分;应用HE染色、组织病理学评分观察牛初乳对小鼠肠炎影响;实时荧光定量PCR(QPCR)监测肠道菌群中常见微生物的含量。结果显示DSS组小鼠体重降低,肠炎评分升高,结肠组织炎性损伤明显,肠道内大肠杆菌含量增加,双歧杆菌、乳酸杆菌含量减少;与DSS组比较,BC+DSS组小鼠体重上升(P＜0.01),肠炎评分降低(P＜0.01),结肠组织炎性损伤显著缓解,三种微生物含量均恢复到正常对照组水平。这表明牛初乳可通过调节肠道菌群改善小鼠溃疡性结肠炎。     

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.     

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.     

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.