Antihyperglycemic Effects of <Emphasis Type="Italic">Platycodon grandiflorum</Emphasis> (Jacq.) A. DC. Extract on Streptozotocin-induced Diabetic Mice     

Zheng J  He J  Ji B  Li Y  Zhang X 《Plant foods for human nutrition (Dordrecht, Netherlands)》2007,62(1):7-11

The root of Platycodon grandiflorum (Jacq.) A. DC has been reported to have a wide range of health benefits in oriental food. This study examined the hypoglycemic effects of Platycodon grandiflorum (Jacq.) A. DC aqueous-ethanol extract (PGE) in streptozotocin (STZ) -induced diabetic ICR mice (STZ diabetic mice) for the first time. The effects of PGE on blood glucose, plasma insulin levels and body weight were investigated. A significant decrease in blood glucose levels was observed after single administration of PGE. Furthermore, Glibenclamide and PGE significantly suppressed the rise in blood glucose after 30 min in the acute glucose tolerance test. Treatment with glibenclamide and PGE resulted in a reduction in blood glucose levels from the 2nd week, and this reduction was maintained until the 4th week of treatment. The body weight changed slightly in glibenclamide and PGE treated mice in comparison with the STZ control group. Plasma insulin levels were increased with glibenclamide treatment in STZ diabetic mice, whereas such effect was not observed with PGE. These results indicated that PGE could induce hypoglycemic effects without stimulating insulin secretion.  相似文献   

非洲猪瘟病毒p30蛋白单克隆抗体制备及线性抗原表位定位     

魏天  王成宇  王凤杰  李忠鹏  张芳毓  张守峰  扈荣良  吕礼良  王永志 《中国农业科学》2022,55(15):3062-3070

【目的】制备非洲猪瘟病毒（African swine fever virus,ASFV）p30蛋白的单克隆抗体（monoclonal antibodies,MAbs）并初步分析其所识别的线性抗原表位,为ASFV及其抗体检测方法的建立及p30蛋白结构和功能的研究奠定基础。【方法】将原核表达并纯化的p30重组蛋白作为免疫原,免疫6—8周龄BALB/c雌鼠,每两周免疫1次,共免疫3次,首次免疫是抗原与等体积的弗氏完全佐剂乳化后免疫,第二次和第三次免疫与等体积的弗氏不完全佐剂乳化,3次免疫后1 w断尾采血,间接酶联免疫吸附试验（ELISA）检测血清抗体效价,选择血清效价最高的小鼠进行加强免疫,3 d后取小鼠脾淋巴细胞与SP2/0骨髓瘤细胞按照4﹕1的比例使用PEG进行常规细胞融合。利用重组p30蛋白作为包被抗原,间接ELISA筛选阳性杂交瘤细胞,有限稀释法进行克隆纯化,直至筛出能够稳定分泌抗体的MAbs。将ASFV接种于猪肺泡巨噬细胞,以筛选的MAbs为一抗、兔抗鼠HRP-IgG为二抗,进行间接免疫荧光试验（IFA）。将感染和未感染ASFV的细胞沉淀处理后进行 SDS-PAGE并转印至硝酸纤维素膜,分别以IFA鉴定为阳性的MAbs上清为一抗、兔抗鼠HRP-IgG为二抗,进行Western blotting分析,筛选获得p30 MAbs。根据已知序列设计引物扩增p30ab与p30bc两段截短基因,其中p30ab代表由第86—153位氨基酸残基的截短体,p30bc代表由第120—187位氨基酸残基的截短体,原核表达部分重叠的截短p30蛋白,最终获得重组蛋白GST-p30ab与重组蛋白GST-p30bc。分别以GST-p30ab和GST-p30bc融合蛋白为包被抗原,以5株MAbs为一抗,以兔抗鼠HRP-IgG为二抗, 通过间接ELISA方法初步定位p30蛋白的抗原表位。【结果】以纯化的重组蛋白为包被抗原,经间接ELISA试验筛选出25株可分泌抗重组 p30蛋白的杂交瘤细胞株。IFA结果显示,5株MAbs（8F4、1D3、1H2、6C3和8E11）与ASFV感染的猪肺泡巨噬细胞IFA 试验呈阳性;Western blotting结果显示,5株MAbs均能够与ASFV感染的细胞呈阳性反应,与未感染病毒的细胞呈阴性反应。试验构建的p30截短体重组蛋白GST-p30ab以可溶和包涵体两种形式表达,而GST-p30bc仅以包涵体形式表达,以两组截短体融合蛋白为包被抗原,通过间接ELISA检测出MAbs 8F4、1H2和6C3与两个重组蛋白均能有效结合,证明MAbs 8F4、1H2和6C3抗原识别区域为两组截短蛋白重叠区域,即第120—153位氨基酸;MAbs 8E11与1D3则只能与GST-p30ab蛋白结合, 证明MAbs 8E11与1D3抗原识别区域为两个重组蛋白的非重叠区域,即第86—119位氨基酸。【结论】本研究可溶性地表达了p30蛋白的第86—153位氨基酸截短体重组蛋白,制备了5株p30 MAbs,定位到2个p30蛋白抗原表位。结合ELISA和IFA,可建立十分可靠的ASFV及其抗体的检测手段。  相似文献   

Immunohistochemical Expression of Potential Therapeutic Targets in Canine Thyroid Carcinoma     

M. Campos  R. Ducatelle  H.S. Kooistra  G. Rutteman  L. Duchateau  I. Polis  S. Daminet 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2014,28(2):564-570

Background

Thyroid carcinoma is a common endocrine tumor in the dog. Local invasive growth frequently precludes surgical excision and, in up to 38% of dogs, the tumor has already metastasized by the time of diagnosis. Therefore, it is important to investigate new treatment modalities that may be useful for the large number of dogs with inoperable tumors or metastatic disease.

Hypothesis/Objectives

To investigate the immunohistochemical expression of potential therapeutic targets in canine thyroid tumors.

Animals

74 dogs with thyroid neoplasia.

Methods

Immunohistochemistry was performed for thyroglobulin, calcitonin, vascular endothelial growth factor (VEGF), p53, cycloxygenase‐2 (cox‐2), and P‐glycoprotein (P‐gp).

Results

Fifty‐four (73%) tumors were classified as follicular cell thyroid carcinomas (FTCs) and 20 (27%) as medullary thyroid carcinomas (MTCs). Eighty percent of FTCs and all MTCs had a high percentage (76–100%) of neoplastic cells immunopositive for VEGF. Thirteen percent of FTCs and 50% of MTCs expressed cox‐2. Seven percent of FTCs and 70% of MTCs expressed P‐gp. No tumor was immunopositive for p53 expression. Expression of VEGF (P = .034), cox‐2 (P = .013), and P‐gp (P < .001) was significantly higher in MTCs compared to FTCs.

Conclusions and Clinical Importance

VEGF is a potential therapeutic target in both FTC and MTC in dogs. Cox‐2 and P‐gp may be useful molecular targets in canine MTC.  相似文献   

牛瑟氏泰勒虫p23-IL-18融合基因的原核表达     

白杨  许应天 《中国兽药杂志》2014,48(7):6-9

为了研发牛瑟氏泰勒虫病基因工程亚单位疫苗,本实验将p23-IL-18融合基因克隆到pET-28a原核表达载体,构建pET-28a-p23-IL-18重组原核表达质粒,经BamHⅠ和EcoRⅠ双酶切鉴定后在37 ℃用IPTG诱导表达,对表达产物进行SDS-PAGE和Western blotting分析。结果表明,成功构建牛瑟氏泰勒虫pET-28a-p23-IL-18原核表达质粒,目的基因在大肠杆菌中成功获得表达,融合蛋白的分子量约为48ku,并被牛瑟氏泰勒虫阳性血清所识别,具有良好的反应原性。本研究结果为牛瑟氏泰勒虫病基因工程亚单位疫苗的制备奠定了基础。  相似文献   

Gonadotropins I and II do not stimulate thein vitro secretion of 17α,20β-dihydroxy-4-pregnen-3-one 20-sulphate by rainbow trout gonads during final sexual maturation     

J. J. Nagler  A. P. Scott  C. R. Tyler  J. P. Sumpter 《Fish physiology and biochemistry》1996,15(2):149-156

Thein vitro secretion of 17,20-dihydroxy-4-pregnen-3-one 20-sulphate (17,20-P-sulphate) and the free steroid 17,20-dihydroxy-4-pregnen-3-one (17,20-P), by rainbow trout (Oncorhynchus mykiss) gonads, in response to gonadotropin (GTH) I and GTH II, were studied during the final stages of sexual maturation. Substantial amounts of 17,20-P-sulphate were produced, by both mature ovaries and testes, indicating considerable 20-hydroxysteroid sulphotransferase (20-HST) activity within these tissues. In the post-ovulatory ovary the level of 17,20-P-sulphate (36.6 ng ml^–1) greatly exceeded that of 17,20-P (8.59 ng ml^–1). The amount of 17,20-P-sulphate produced in incubations of both mature ovary and testes was unaffected by either GTH I or GTH II treatment at physiological concentrations up to 100 ng ml^–1. Similarly, incubations of maturing ovary and testes, treated with GTH I or GTH II, in the presence of added 17,20-P at 100 ng ml^–1 of medium, produced levels of 17,20-P-sulphate that were similar to those of the controls. In incubations of mature ovarian follicles at the stages of germinal vesicle breakdown and preovulation, both GTHs significantly stimulated secretion of 17,20-P, although GTH II was always more potent than GTH I. GTH II significantly elevated the levels of 17,20-P in testicular incubations from mature males more than 4-fold relative to GTH I and controls, which did not differ from one another.In conclusion, 20-HST, the enzyme responsible for the sulphate conjugation of 17,20-P, was found to be active in the ovaries and testes of rainbow troutin vitro. However, the levels of this enzyme do not appear to be regulated by either GTH I or GTH II.  相似文献   

Hydrogen sulfide ameliorates high glucose-induced endothelial cell senescence by suppressing oxidative stress     

SONG Zhi-ming  WANG Min  LIU Yong  HAO Bao-shun  NIU Hai-ming  LIU Ding-hui  YU Shu-jie  ZHOU Bin  WU Lin  YU Xian-guan  LING Ye-sheng  PENG Pei  ZHU Jie-ming  CHEN Lin  QIAN Xiao-xian 《园艺学报》2014,30(8):1345-1350

AIM：To explore the effect of hydrogen sulfide on the senescence of human umbilical vein endothelial cells (HUVECs) induced by high glucose. METHODS：Senescence model was established by treating HUVECs with 33 mmol/L glucose for 48 h. The parameters were detected to demonstrate the effect of hydrogen sulfide on senescence and the mechanism involved was also investigated. RESULTS：In the cells treated with high glucose, the proliferation was attenuated with a higher number of senescence-associated β-galactosidase (SA-β-Gal) positive cells, and plasminogen activator inhibitor 1 (PAI-1) protein expression, malondialdehyde (MDA) production and NF-κB p65 activity were increased significantly, but the expression of superoxide dismutase 1 (SOD1) was decreased. However, the cell number and SOD1 expression were increased, and the number of SA-β-Gal positive cells, PAI-1 protein expression, MDA production and the activity of NF-κB p65 were decreased after sodium hydrosulfide (100 and 200 μmol/L) treatment.CONCLUSION：Exo-genous hydrogen sulfide prevents HUVECs against high glucose-induced senescence by suppressing oxidative stress and NF-κB p65 activity.  相似文献   

p53调控Nfkbiz基因表达的研究     

杨逸凡  杜立中  朱紫祥  李满  魏建超  史子学  邵东华  李蓓蓓  马志永 《中国动物传染病学报》2014,(3):61-68

核因子-κB（NF-κB）在炎症反应中起着关键的促进作用,并且受其他多种因素的调节。Nfkbiz基因编码的IκB？蛋白是IκB家族成员之一,在炎症反应中起着调节NF-κB的作用。本研究采用荧光实时定量PCR分析Nfkbiz基因的表达情况,发现在p53特定刺激源作用下Nfkbiz基因mRNA丰度显著降低。分析Nfkbiz基因序列,发现其基因中存在可能的p53反应元件,通过构建荧光素酶报告质粒及双荧光素酶报告试验和染色质免疫共沉淀试验,证明Nfkbiz基因中含有p53反应元件并且受p53蛋白调节。以上结果初步证明Nfkbiz是受p53蛋白负向调控的靶基因。  相似文献   

Gene Expression in Black Tiger Prawns Following Intramuscular Injection of β-gal Plasmid     

Z.H. Sulaiman  R.H. Ming Chan  P.M. Simanjuntak 《Aquaculture International》1999,7(5):333-340

Gene expression in black tiger prawns (Penaeus monodon) was studied following intra-muscular injection of CMVGal plasmid into the second abdominal segment. We used an in situ staining technique to detect -gal expression in one- and three-month-old injected prawns. We found that only one of the three-month-old prawns expressed the marker gene (2 days after injection), and the site of expression was confined to the sixth abdominal segment away from the injection site. We repeated the experiment on a new batch of three-month-old prawns, but using fluorometric determination technique. This time we found that -gal expression was detected (6/42) at the site of injection after 2, 7, and 14 days. In two other test samples, transgene expression was detected in the sixth abdominal segment only, further confirming the possibility of injected DNA dispersal. The results of the study also suggest that direct gene transfer is a feasible technique in black tiger prawns.  相似文献   

Effects of dietary lipids on body composition and liver function in juvenile red drum, Sciaenops ocellatus   总被引：2，自引：0，他引：2  

S.R. Craig  B.S. Washburn  D.M. Gatlin III 《Fish physiology and biochemistry》1999,21(3):249-255

This study was conducted to evaluate the effects of different concentrations of dietary lipids on body composition and liver function in juvenile red drum, Sciaenops ocellatus. Diets were formulated to contain 40% crude protein from solvent-extracted menhaden fish meal and 0, 7, 14 or 21% lipid from menhaden fish oil. The basal diet, without supplemental fish oil, contained lipid at 0.4% of dry weight. The diets were fed to groups of 25 juvenile red drum initially averaging 7.3 ± 0.18 g fish^–1 in a recirculating culture system for 8 weeks and weight gain was recorded. After an additional 8 weeks, 16 fish from each treatment were sacrificed and the following measurements were recorded: hepatosomatic index (HSI), intraperitoneal fat (IPF) ratio, and liver -tocopherol, malondialdehyde (MDA) formation, and cytochrome P-4501A activity (measured as 7-ethoxyresorufin O-deethylase (EROD) activity). The activity of alanine and aspartate aminotransferases and concentrations of -tocopherol also were measured in plasma.Weight gain was significantly (p<0.05) affected by dietary lipid concentration, with values ranging from 361% of initial weight for fish fed the basal diet to 527% of initial weight for fish fed the diet containing 7% lipid. The HSI and IPF ratio values also were significantly affected by lipid with the lowest values recorded for fish fed the basal diet and the highest values observed in fish fed the diet containing 21% lipid. Increasing dietary lipid significantly increased oxidative stress as reflected in reduced -tocopherol in liver and plasma and increased MDA formation in the liver, although no overt pathological signs were observed. These findings suggest that lipid concentrations between 7 and 14%, when the diet contains 60 IU vitamin E kg^–1, are likely to limit oxidative stress and result in normal physiological responses of red drum.  相似文献   

密植浒苔对冬季露天池塘池底水温、酸碱度和溶解氧的影响效应     

富裕  ;Fatou Diouf  ;陈汉春  ;袁向阳  ;任夙艺  ;王志铮 《浙江水产学院学报》2014,(5):389-393

借助环境数据实时采集装置,于2014年1月5日至1月22日连续开展了冬季露天池塘培植浒苔对池底水温、酸碱度和溶解氧的实时监测工作。结果表明：（1）密植浒苔可在不改变池底水温峰值期时辰数的基础上,提前1个时辰到达峰值,在不影响到达谷底时辰的同时,使低谷期缩短了1个时辰,并由此延长了水温由峰到谷的转换时长;（2）密植浒苔可保持池底p H值的基本稳定,并在不改变池底p H值时段变化节律的基础上,使日均p H值由9.3下调至8.6;（3）密植浒苔在对池底DO值的时序变化影响上与池底水温相仿,并使日均DO值由10.61mg/L下调至6.80mg/L。  相似文献