Effect of p19ARF on growth and apoptosis of leukemia cells

AIM:To study the effect of p19ARF on the biological behavior of human leukemia cells. METHODS:p19ARF was cloned in eukaryotic expression vector pcDNA3.1 and transferred into INK4a/ARF locus-deficient leukemia cells HEL and K562. The changes in biological characteristics of the two p19ARF-transfected cells were observed.RESULTS:The growth of the p19ARF-transfected HEL cells was significantly inhibited compared with the vector-transfected cells; Cell cycle analysis showed that the expression of foreign p19ARF gene resulted in G1 phase cell cycle arrest and apoptosis cell death in some of HEL cells. However, p19ARF had no marked effects on the growth of K562 cells with p53 gene mutation and did not induce apoptosis in K562 cells.CONCLUSION:p19ARF suppressed the growth of leukemia cells by p53-dependent pathway.     

Expression of Survivin and its relationship to prognosis in human hepatocellular carcinoma

AIM: To study the expression of Survivin and its relationship to prognosis in human hepatocellular carcinoma(HCC).METHODS: The expression of Survivin protein in 83 cases of HCC and their neighboring noncancerous tissues was detected by immunohistochemistry.The expression of Survivin mRNA in 11 cases of HCC and their neighboring noncancerous tissues was detected by semi-quantitative RT-PCR.RESULTS: Survivin protein expression was detected in 53 of 83 (63.9%) HCC tissues and 21 of 53 (39.6%) corresponding noncancerous tissues.22 of 53 Survivin-positive HCCs (41.5%) showed punctate nuclear staining in HCC cells,24 of 53 Survivin-positive HCCs (45.3%) showed cytoplasmic staining in HCC cells and the remaining 7 showed both nuclear and cytoplasmic staining.The positive rates of nuclear Survivin expression in cases of envelope invasion or tumor metastasis were significantly higher than those in cases without envelope invasion or tumor metastasis (P<0.05).The patients with positive nuclear Survivin expression had the higher chance of poor prognosis than those with negative nuclear Survivin expression (P<0.01).Survivin mRNA was detected in all the 11 HCC and 5 of 11 (45.5%) neighboring noncancerous tissues.CONCLUSION: There is a high expression of Survivin in HCC and positive nuclear Survivin may play an important role in malignant biological behavior and prognosis of HCC.     

Research on Antimicrobial Resistance Characteristics of Enterococcus spp. from Companion Animals in Beijing

In order to study the tolerance of Enterococcus spp.from companion animals to common antibiotics in Beijing,320 samples were collected from dogs and cats in a pet hospital in Beijing from May 2015 to January 2016.The strains were isolated by selective culture medium and identified by VITEK-2 Compact 60.The sensitivity test of 11 kinds of common antibiotics in 8 categories was carried out for the isolated and identified strains.At the same time,the high-level gentamicin and high-level streptomycin resistance test were carried out to analyze the drug resistance and multiple drug resistance of Enterococcus.A total of 318 strains of non repetitive enterococci with different colony morphology and biochemical characteristics were obtained.The isolation rates of Enterococcus faecalis (49.06%) and Enterococcus faecium (29.87%) were higher.The resistance rates of Enterococcus to high-level gentamicin and high-level streptomycin were 39.94% and 43.40%,respectively.The resistance rates to tetracycline (78.62%),erythromycin (67.30%) and quinoproptin dapfoptin (43.71%) were very high.All of them were sensitive to tigecycline,and the non-resistant rate to vancomycin and nitrofurantoin was also high (98.74%).Enterococcus tested showed different degrees of resistance to class 1 to class 7 antibiotics,and the multidrug resistance was common,and the multidrug resistance rate was as high as 57.23%.The number of resistant antibiotics was the highest (n=65).A total of 44 different drug resistance profiles were obtained.Among them,erythromycin-quinoptin-daptoptin-tetracycline-high level aminoglycoside resistant spectrum accounted for the highest proportion (45/318,14.15%).This study confirmed that Enterococcus spp.isolated from companion animals in Beijing was highly resistant to common antibiotics,and multidrug resistance was prevalent.Therefore,it was necessary to strengthen the monitoring of drug resistance.     

伪狂犬病病毒变异株gC抗体间接ELISA检测方法的建立及初步应用

本研究旨在建立变异伪狂犬病病毒FJ-2012株gC蛋白抗体间接ELISA检测方法,掌握不同猪场猪群的gC抗体水平情况。将FJ-2012株gC基因连接至pCzn1载体,转染至Arctic express(DE3)感受态细胞,IPTG诱导表达产物经SDS-PAGE和Western blot验证,通过矩阵法试验、临界值的确定、特异性试验、重复性和敏感性试验,建立PRV-gC抗体间接ELISA检测方法,并对源于不同猪场的280份血清进行PRV-gC抗体检测。结果表明,成功表达获得FJ-2012株pCzn1-gC重组蛋白;建立的间接ELISA方法的最佳抗原包被浓度为10 μg·mL^-1和最佳样品血清稀释比例为1∶50,阴性和阳性判定的OD_{650 nm}临界值为0.406~0.438,介于之间的判定为可疑;临床检测结果显示,120份盲样猪血清PRV-gC抗体阳性率为91.67%、PRV-gB抗体阳性率为95.00%,两种抗体检测结果的整体符合率为95.83%;PR不稳定的猪场血样PRV-gC抗体阳性率为96.25%(77/80),而PRV已净化的种猪场血样PRV-gC抗体阳性率为78.75%(63/80)。因此,建立的PRV变异株gC抗体间接ELISA检测方法为临床猪血清抗体检测提供了特异、灵敏和稳定的技术,也为开展变异PRV血清学调查奠定基础。     

高职“五年一贯制”学生逆反心理的探析

"五年一贯制"的学生的逆反心理的产生的原因很多,其校正过程与其班主任的教育引导有很大的关系,所以班主任应因势利导,作好学生逆反心理的转化工作。     

沙棘果油的毒性实验研究

沙棘果油以15ml/kg,30ml/kg及60ml/kg分组给小白鼠ig;10ml/kg,20ml/kg及40 ml/kg分组给大白鼠ig.观察7 d,皆未测出LD50.给小白鼠SC10ml/kg,20 ml/kg;给兔背部完整或破损皮肤涂2.5 ml/kg及10ml/kg,观察7 d,未见明显中毒症状.按2ml/kg,5 ml/kgg及10ml/kg每天分组给大白鼠ig,连续90 d.给油组鼠的体重,血液学检查,血液生化学检查等,与对照组比较皆无显著性差异(P＞0.05).给油组鼠的心、肝、脾、肺、肾、肾上腺、胸腺、睾丸、子宫卵巢等器官肉眼观察、及病理切片组织形态光镜检查,均未发现明显损害性病理改变,与对照组比较,未发现明显差异.对豚鼠皮肤,兔眼睛及阴道局部应用,未见明显刺激反应.豚鼠皮肤过敏试验,未见过敏反应.     

炼铁乡桉树林分改造参试树种生长调查分析

根据云南省炼铁乡的自然环境条件，选择加州李、中林美荷杨、圣诞树、桤木、慈竹、香竹、梅树等7个树种进行造林试验。通过对树种主要指标的测试和数学分析，初步选择出该地区桉树林分改造的参试树种继续观测和试验。     

应用双向电泳分析2株不同毒力的副猪嗜血杆菌蛋白质表达的差异

副猪嗜血杆菌是猪常见细菌性疾病的病原之一,临床表现为多发性纤维素性浆膜炎、关节炎和脑膜炎。运用蛋白质组学技术对不同毒力的副猪嗜血杆菌(Haemophilus parasuis)进行比较研究,明确差异蛋白质信息,对揭示其致病机制具有重要意义。本试验通过应用建立的双向电泳方法对选取同为4型的2株不同H.parasuis临床分离株(菌株A和菌株B)进行蛋白质组学研究,二者相互比较,共鉴定出10个差异表达蛋白质,其中菌株A有3个蛋白质表达量为增加,分别是谷氨酰tRNA合成酶、半胱氨酸合成酶和假设蛋白;菌株B有7个蛋白质表达量为增加,包括Yae T蛋白(omp85)、CTP合酶、天冬氨酰tRNA合成酶(有3个)、尿苷激酶、SecB蛋白。差异蛋白质的功能主要集中在蛋白质合成、氨基酸合成和嘧啶合成。研究结果进一步丰富了H.parasuis的毒力因子和致病机制的相关信息,也为今后开展H.parasuis免疫蛋白质组学研究奠定了基础。     

Diagnostic Report of Clostridium chauvoei Infection of Cattle in Yanbian of Jinlin Province

The study was aimed to rapidly diagnose two ill and dead cattle in three bay town of Yanji city in Jilin province.We collected liver,spleen,heart,muscle and other tissues to diagnose Clostridium chauvoei infection by epidemiological investigation,clinical autopsy,microscopic obsevation,bacterial culture observation,biochemical experiment,molecular biology diagnosis and animal experiment.The muscles of the ill and dead cattle contacted with crepitus,and the section had a lot of blood and bubble flowing out,microscopic obsevation found bacillus with obtuse at both ends and spores,the bottom of the tube had loose and white precipitate in the broth,biochemical experiments showed that the isolate had acid-production and gas-production reactions which was unique to anaerobic bacteria causing Clostridium chauvoei infection,and 501 bp fragment was amplified by PCR.The results showed that two ill and dead cattle were casued by Clostridium chauvoei infection in Yanbian area,and proved the existence of Clostridium chauvoei.The test separated Clostridium chauvoei Yanbian strain for the frist time,which provided important reference basis for the prevention and treatment of Clostridium chauvoei infection,and laid the important foundation for the further study of drug and immune preventions of this disease.     

Shenmai injection improves insulin resistance in 3T3-L1 cells

AIM: To discuss the effect of Shenmai injection on insulin resistance (IR) in 3T3-L1 cells and its mechanisms. METHODS: 3T3-L1 preadipocytes were induced by chemical reagents to differentiate into fully differentiated adipocytes. Oil red O staining was used to detect the differentiation level of the adipocytes. The insulin-resistant 3T3-L1 cell model was demonstrated using insulin, which was confirmed by glucose concentration in cell supernatant. The IR cell model was given 10 μmol/L rosiglitazone, 25 and 50 g/L Shenmai injection and normal saline for comparison. MTT assay was used to assess the cell activity of 3T3-L1 cells which was treated with drugs for 8, 16, 24 and 36 h. Glucose oxidase method was used to detect the glucose concentration in the cell supernatant at 8, 16 and 24 h. The protein levels of glucose transporter-4 (GLUT4), phosphatidylinositol 3-kinase (PI3K), AKT and p-AKT were determined by Western blot. RESULTS: 3T3-L1 adipocytes were successfully induced as shown by the positive oil red O staining. The IR cell model was demonstrated, and glucose concentration in the cell supernatant after treatment with Shenmai injection showed that Shenmai injection reduced the IR in 3T3-L1 cell model. The protein levels of GLUT4, PI3K and p-AKT increased compared to control group. CONCLUSION: Shenmai injection reduces the IR in 3T3-L1 cell model, which functions by increasing the protein levels of GLUT4, PI3K and p-AKT.