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1.
为了探究黄藤素的体外细胞毒性,采用不同浓度的黄藤素作用山羊子宫内膜上皮细胞(EECs)后,采用MTT法绘制细胞生长曲线;倒置显微镜下观察细胞生长状态;瑞氏-姬姆萨染色法观察细胞形态学变化并通过测定乳酸脱氢酶(LDH)含量检测细胞膜的完整性。结果显示,当黄藤素的浓度为10~227 μg/mL时,黄藤素对EECs有促增殖的作用;当浓度大于227 μg/mL时出现细胞增殖抑制现象,药物对细胞的半数增殖抑制率(IC50)为360 μg/mL;瑞氏-姬姆萨染色结果显示,100、200 μg/mL的黄藤素作用于EECs时均未对其细胞核、细胞浆的形态产生影响;LDH结果显示,药物浓度≥250 μg/mL时细胞LDH的释放量极显著高于正常细胞组(P<0.01)。综上所述,黄藤素作用于山羊EECs后呈现一定的毒性作用,且呈现显著的剂量依赖效应,作用于EECs的安全剂量范围为≤250 μg/mL。  相似文献   

2.
《中国兽医学报》2016,(9):1572-1576
通过研究不同浓度的咖啡酸对脂多糖(LPS)诱导的犬子宫内膜上皮细胞活率下降的影响,探讨咖啡酸的抗炎作用效果,为宠物临床治疗犬子宫内膜炎的新药研发提供依据。根据LPS剂量筛选试验确定LPS诱导炎症模型的浓度;通过咖啡酸药物毒性试验,根据抑制率IR≤10%,选择低、中、高3个咖啡酸剂量组;用噻唑蓝比色法(MTT)测定不同剂量组的咖啡酸对LPS诱导的细胞活率下降的影响。结果显示,50mg/L LPS作用于子宫内膜上皮细胞12h能够引起细胞活率的显著下降(P0.05);咖啡酸作用于子宫内膜上皮细胞12h后,与对照组相比,1μg/L~200mg/L组细胞活率均没有显著差异(P0.05),且IR10%,500 mg/L组有极显著性差异(P0.01),且IR40%;作用24h后,1μg/L~25mg/L组细胞活率没有显著差异(P0.05),IR10%,50~100mg/L组有显著差异(P0.05)且IR10%,200、500mg/L组有极显著差异(P0.01),IR50%;咖啡酸中剂量组(50mg/L)和咖啡酸高剂量组(100mg/L)对LPS引起的细胞的活率具有明显的提高作用。结果表明,咖啡酸对LPS诱导的子宫内膜细胞活力下降具有一定的保护作用,且在安全范围内,随着药物浓度升高,保护作用增强。  相似文献   

3.
本试验通过传代培养奶牛子宫内膜上皮细胞,以0.1μg/mL、1μg/mL、10μg/mL、50μg/mL、100μg/mL、500μg/mL、1000μg/mL 7个LPS浓度梯度刺激细胞,分别于刺激后12h、24h、36h 3个时间段做MTT细胞毒检测,同时进行细胞形态学观察,研究LPS对子宫内膜上皮细胞的毒性作用。结果发现:0.1μg/mL、1μg/mLLPS组细胞形态学无明显改变;10μg/mL、50μg/mL、100g/mL LPS组细胞形态学与对照组相比有明显改变,500μg/mL、1000μg/mL LPS组细胞发生裂解、死亡。MTT试验结果表明LPS对细胞的抑制作用呈浓度与时间依赖关系,随着LPS浓度的增加和刺激时间的延长,细胞抑制率明显增加,细胞活力明显降低。  相似文献   

4.
本试验旨在探究孕酮对奶牛子宫内膜上皮细胞MAPK信号通路的影响。选取与奶牛产后体内相似浓度的孕酮作用于奶牛子宫内膜上皮细胞,模拟产后子宫内分泌环境,检测MAPK通路关键蛋白表达的变化。结果发现,不同浓度孕酮均未对细胞活性产生影响,且20 ng/m L浓度范围内的孕酮对奶牛子宫内膜上皮细胞无毒性作用。用0、1、3、5 ng/m L孕酮处理30 min后,并未对子宫内膜上皮细胞ERK、JNK和P38的磷酸化产生显著作用,且不同浓度孕酮之间也没有显著差异。由此得出结论,外源性孕酮不能通过直接抑制奶牛子宫内膜上皮细胞MAPK的活化起抗炎效应。  相似文献   

5.
本试验旨在体外分离培养兔子宫内膜细胞(腺上皮细胞和基质细胞)和平滑肌细胞,并且探讨纯化子宫内膜上皮细胞和基质细胞的方法。用差速离心法和差速贴壁法获得纯化的子宫内膜上皮细胞和基质细胞,用滤网法分离得到子宫平滑肌细胞,分别在添加20%胎牛血清(FBS)、100μg/mL牛胰岛素、63.5 nmol/L雌二醇(E2)、7.14nmol/L孕酮(P4)的DMEM/F12(1∶1)培养液和37℃5%CO2的饱和湿度条件下培养;用免疫组化和免疫荧光法鉴定细胞并检测分离的细胞纯度。结果表明用该方法成功地分离培养了兔子宫内膜上皮细胞和基质细胞,且2种细胞纯度都达98%左右,基质细胞出现蜕膜化;子宫平滑肌细胞的纯度为97%左右,而且出现横纹状的生长。本研究表明子宫内膜细胞和平滑肌细胞能够在体外成功培养;在含20?S、100μg/mL牛胰岛素、63.5 nmol/L E2、7.14 nmol/L P4的DMEM/F12(1∶1)培养液和37℃5%CO2饱和湿度的培养条件下最有利于体外培养的子宫内膜细胞和平滑肌细胞的生长。  相似文献   

6.
为探究百里香酚治疗子宫内膜炎的抗炎活性,利用脂多糖(LPS)诱导山羊子宫内膜上皮细胞(gEECs)建立炎性模型,以百里香酚进行干预。采用NO试剂盒和酶联免疫吸附试验(ELISA)分别检测百里香酚对LPS诱导的NO和细胞因子分泌的影响;利用实时荧光定量PCR(RT-PCR)检测百里香酚对LPS诱导的IL-1β、TLR4和NF-κB基因转录的影响。结果显示:不同剂量的百里香酚(100、50、25μg·mL~(-1))能不同程度地抑制LPS诱导的炎性模型细胞TNF-α、IL-1β、IL-6、NO和PGE2的分泌,亦可降低炎性模型细胞IL-1β、TLR4和NF-κB的基因转录水平(P0.01)。结果表明,百里香酚可抑制LPS诱导gEECs炎性模型的细胞因子表达水平,具有显著的抗炎活性,其作用机制可能与抑制TLR4/NF-κB信号通路相关。  相似文献   

7.
为研究丹参水提物(SME)对子宫内膜上皮细胞炎症模型中基质金属蛋白酶-2(MMP-2)表达的影响,体外培养山羊子宫内膜上皮细胞(EEC),利用内毒素(脂多糖,LPS)诱导细胞增殖和产生TNF-α,建立EEC炎症模型。然后给予高、中、低不同剂量SME干预。应用MTT比色法检测细胞活力,ELISA试剂盒检测TNF-α含量,实时荧光定量PCR检测细胞MMP-2mRNA表达变化。结果表明,5μg/mL LPS作用EEC 12h时促增殖作用显著(P0.01);SME可显著促进EEC炎症模型中MMP-2的表达,以中、高浓度组的作用较为明显。说明SME对LPS引起的子宫内膜炎症反应有一定的保护作用。  相似文献   

8.
本研究旨在探究瘦素对绵羊子宫内膜上皮细胞中脂代谢过程和胚胎植入关键基因表达的影响。在发情期采集成年健康绵羊的子宫内膜上皮组织,利用组织块法培养、胰酶差时消化法纯化及角蛋白鉴定分离绵羊子宫内膜上皮细胞。经浓度筛选后,选取最适瘦素浓度250μg/mL处理绵羊子宫内膜上皮细胞,并进行脂滴检测,采用RT-qPCR法检测瘦素对胚胎植入关键基因的影响。结果显示:在250μg/mL瘦素处理下,子宫内膜上皮细胞增殖能力提升,出现脂滴聚集现象,脂代谢相关因子脂肪细胞脂肪酸结合蛋白基因表达升高(P<0.05),过氧化物酶增殖激活受体γ基因表达升高(P<0.05);胚胎植入相关因子白血病抑制因子基因表达升高(P<0.05),整合蛋白β-3(P<0.01)、白介素-6(P<0.05)基因表达降低。本研究结果表明,250μg/mL的瘦素处理绵羊子宫内膜上皮细胞后会显著增强其细胞增殖能力,对细胞的脂代谢有显著的上调作用,对绵羊胚胎植入关键基因有显著影响,提示瘦素在绵羊胚胎附植过程中通过影响脂代谢进程影响胚胎附植过程。  相似文献   

9.
本试验旨在研究体外培养中硒(Se)对凡纳滨对虾(Litopenaeus vannamei)肝胰腺细胞抗氧化能力的影响以及对黄曲霉毒素B1(AFB1)毒性的缓解作用。在体外培养肝胰腺细胞的基础上,设定含不同浓度(0、0.15、0.30、0.45、0.60、0.75μg/m L)Se的培养液,研究Se对肝胰腺细胞抗氧化能力的影响;另用浓度为1 600μg/L的AFB1侵染细胞,探究Se对AFB1毒性的缓解作用。结果表明:当Se浓度处于0~0.45μg/m L时,随着Se浓度的增加,丙二醛(MDA)含量呈现降低趋势,但各个浓度之间并没有显著差异(P0.05),过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)活力随着Se浓度的增加而增强,Se浓度为0.45μg/m L时GSH-Px活力显著高于其他浓度(除0.30μg/m L)时(P0.05);当Se浓度处于0.45~0.75μg/m L时,随着MDA含量升高的同时CAT、GSH-Px活力下降。当细胞受到浓度为1 600μg/L的AFB1侵染后,随着培养时间的增加,MDA含量表现一直升高的趋势,同时CAT、GSH-Px活力都有一定程度的下降;当细胞受到浓度为1 600μg/L的AFB1和浓度为0.45μg/m L的Se共同作用后,随着培养时间的增加,MDA含量有一定程度的改善,CAT、GSH-Px活力得到一定程度的恢复。由此可见,在体外培养条件下,浓度为0.45μg/m L的Se有利于提高凡纳滨对虾肝胰腺细胞的抗氧化能力,且在一定程度上能缓解AFB1对肝胰腺细胞的毒性作用。  相似文献   

10.
探讨黄芩苷体外抗马立克氏病病毒(MDV)作用,应用细胞病变观察法(CPE)观察MDV对鸡胚成纤维细胞(CEF)的影响,采用Reed-Muench法计算MDV细胞半数感染量(TCID50),以CEF为宿主细胞,应用四甲基偶氮唑蓝(MTT)比色法从黄芩苷对MDV的增殖抑制、感染阻断以及直接灭活3个给药途径探讨黄芩苷抗MDV作用。结果显示:MDV对CEF的TCID50为10-4.5/0.1 m L;3种给药途径下,黄芩苷浓度在312.5~78.13μg/m L范围都具有较好的抗MDV感染作用,并且感染阻断途径下黄芩苷浓度为312.5~156.25μg/m L时能够促进细胞的生长,直接灭活途径下黄芩苷浓度为312.5μg/m L时能够促进细胞的生长。其中感染阻断与直接灭活途径给药比增值抑制途径给药的抗MDV感染作用效果要好,直接杀灭途径给药时的病毒抑制率最高。  相似文献   

11.
试验选用96头平均体重14.82 kg左右的杜×长×大断奶仔猪,随机分成4组,每组3栏,每栏8头(公母各半)。对照组饲喂基础日粮,试验1、2、3组分别添加1%80目白术、0.2%白术多糖和1%微米白术。试验期30 d。结果表明:在生长性能方面,与对照组相比,1%微米白术添加组可显著提高日增重(P0.05)、降低饲料增重比和腹泻率,而且效果优于1%80目白术组和0.2%白术多糖组,在肠道形态和肠道微生态区系方面,与对照组相比,日粮添加1%80目白术、0.2%白术多糖、1%微米白术均可不同程度的提高十二指肠和空肠的绒毛高度,加深十二指肠和空肠的隐窝深度,并且增加肠道微生态区系的多样性,其中以1%微米白术添加组的效果最佳。  相似文献   

12.
Sissay, M.M., Uggla, A. and Waller, P.J., XXXX. Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia. Tropical Animal Health and Production, XXXX. A 2-year abattoir survey was carried out to determine the prevalence, abundance and seasonal incidence of gastro-intestinal (GI) nematodes and trematodes (flukes) of sheep and goats in the semi-arid zone of eastern Ethiopia. During May 2003 to April 2005, viscera including liver, lungs and GI tracts were collected from 655 sheep and 632 goats slaughtered at 4 abattoirs located in the towns of Haramaya, Harar, Dire Dawa and Jijiga in eastern Ethiopia. All animals were raised in the farming areas located within the community boundaries for each town. Collected materials were transported within 24 h to the parasitology laboratory of Haramaya University for immediate processing. Thirteen species belonging to 9 genera of GI nematodes (Haemonchus contortus, Trichostrongylus axei, T. colubriformis, T. vitrinus, Nematodirus filicollis, N. spathiger, Oesophagostomum columbianum, O. venulosum, Strongyloides papillosus, Bunostomum trigonocephalum, Trichuris ovis, Cooperia curticei and Chabertia ovina), and 4 species belonging to 3 genera of trematodes (Fasciola hepatica, F. gigantica, Paramphistomum {Calicohoron} microbothrium and Dicrocoelium dendriticum) were recorded in both sheep and goats. All animals in this investigation were infected with multiple species to varying degrees. The mean burdens of adult nematodes were generally moderate in both sheep and goats and showed patterns of seasonal abundance that corresponded with the bi-modal annual rainfall pattern, with highest burdens around the middle of the rainy season. In both sheep and goats there were significant differences in the mean worm burdens and abundance of the different nematode species between the four geographic locations, with worm burdens in the Haramaya and Harar areas greater than those observed in the Dire Dawa and Jijiga locations. Similar seasonal variations were also observed in the prevalence of flukes. But there were no significant differences in the prevalence of each fluke species between the four locations. Overall, the results showed that Haemonchus, Trichostrongylus, Nematodirus, Oesophagostomum, Fasciola and Paramphistomum species were the most abundant helminth parasites of sheep and goats in eastern Ethiopia.  相似文献   

13.
1-(2-Chloroethyl)3-cyclohexyl-1-nitrosourea (CCNU) is an alkylating agent in the nitrosourea subclass. A prospective evaluation of CCNU was done to determine the maximally tolerated dosage of CCNU in tumor-bearing cats. Response data were obtained when available. Twenty-five cats were treated with CCNU at a dosage of 50-60 mg/m3 body surface area. Complete hematologic data were available for 13 cats. Neutropenia was the acute dose-limiting toxicity. The median neutrophil count at the nadir was 1,000 cells/microL (mean, 2,433 cells/microL; range, 0-9,694 cells/microL). The time of neutrophil nadir was variable, occurring 7-28 days after treatment, and counts sometimes did not return to normal for up to 14 days after the nadir. Based on these findings, a 6-week dosing interval and weekly hematologic monitoring after the 1st treatment with CCNU are recommended. The nadir of the platelet count may occur 14-21 days after treatment. The median platelet count at the nadir was 43,500 cells/microL. No gastrointestinal, renal, or hepatic toxicities were observed after a single CCNU treatment, and additional studies to evaluate the potential for cumulative toxicity should be performed. Five cats with lymphoma and 1 cat with mast cell tumor had measurable responses to CCNU. Phase II studies to evaluate antitumor activity should be completed with a dosing regimen of 50-60 mg/m3 every 6 weeks.  相似文献   

14.
Continuous rate infusion (CRI) of furosemide in humans is considered superior to intermittent administration (IA). This study examined whether furosemide CRI, compared with IA, would increase diuretic efficacy with decreased fluid and electrolyte fluctuations and activation of the renin-angiotensin-aldosterone system (RAAS) in the horse. Five mares were used in a crossover-design study. During a 24-hour period, each horse received a total of 3 mg/kg furosemide by either CRI (0.12 mg/kg/h preceded by a loading dose of 0.12 mg/kg IV) or IA (1 mg/kg IV q8h). There was not a statistically significant difference in urine volume over 24 hours between methods; however, urine volume was significantly greater after CRI compared with IA during the first 8 hours ([median 25th percentile, 75th percentile]: 9.6 L [8.9, 14.4] for CRI versus 5.9 L [5.3, 6.0] for IA). CRI produced a more uniform urine flow, decreased fluctuations in plasma volume, and suppressed renal concentrating ability throughout the infusion period. Potassium, Ca, and Cl excretion was greater during CRI than IA (1,133 mmol [1.110, 1,229] versus 764 mmol [709, 904], 102.7 mmol [96.0, 117.2] versus 73.3 mmol [65.0, 73.5], and 1,776 mmol [1,657, 2.378] versus 1,596 mmol [1,457, 1,767], respectively). Elimination half-lives of furosemide were 1.35 and 0.47 hours for CRI and IA, respectively. The area under the excretion rate curve was 1,285.7 and 184.2 mL x mg/mL for CRI and IA, respectively. Furosemide CRI (0.12 mg/kg/h) for 8 hours, preceded by a loading dose (0.12 mg/kg), is recommended when profound diuresis is needed acutely in horses.  相似文献   

15.
Genetic variations in chromosome Y are enabling researchers to identify paternal lineages, which are informative for introgressions and migrations. In this study, the male‐specific region markers, sex‐determining region‐Y (SRY), amelogenin (AMELY) and zinc finger (ZFY) were analysed in seven Turkish native goat breeds, Angora, Kilis, Hair, Honaml?, Norduz, Gürcü and Abaza. A SNP in the ZFY gene defined a new haplotype Y2C. All domestic haplogroups originate from Capra aegagrus, while the finding of Y1A, Y1B, Y2A and Y2C in 32, 4, 126 and 2 Turkish domestic goats, respectively, appears to indicate a predomestic origin of the major haplotypes. The occurrence of four haplotypes in the Hair goat and, in contrast, a frequency of 96% of Y1A in the Kilis breed illustrate that Y‐chromosomal variants have a more breed‐dependent distribution than mitochondrial or autosomal DNA. This probably reflects male founder effects, but a role in adaptation cannot be excluded.  相似文献   

16.
The epidemiology of H. placei and of other gastrointestinal nematodes in yearling dairy cattle was examined on two farms in Kiambu District, central Kenya during each of 13 one-month periods from April 1993 to April 1994. On each farm, 32 newly weaned dairy calves were given a single dose of albendazole and then placed on experimental pastures. Twelve of the animals were designated for bi-monthly slaughter (n = 2) and analysis of worm population characteristics and 20 were designated for blood and faecal collection and for weighing. Two parasite-free tracer calves were grazed alongside the weaner calves each month throughout the study period and were also slaughtered for analysis of worm populations. Faecal egg counts, haematological and serum pepsinogen determinations, herbage larval counts, and animal live weight changes were recorded monthly. The study revealed that Haemonchus placei, Trichostrongylus axei, Cooperia spp. and Oesophagostomum radiatum were responsible for parasitic gastroenteritis and that H. placei was the predominant nematode present in the young cattle on both farms. Faecal egg counts from resident cattle and necropsy worm counts revealed that pasture larval levels were directly related to the amount of rainfall. The total worm burdens in the animals were highest during the rainy season (March–June and October–December) and lowest during the dry seasons (July–September and January–February). The very low recovery of immature larvae of H. placei from the tracer calves indicated that arrested development is not a feature of the life cycle of this parasite in central Kenya. The maintenance of the parasite population depended on continuous cycling of infection between the host and the pasture. The agroclimatic conditions of the study area were such that, in general, favourable weather conditions for the development and survival of the free-living stages of gastrointestinal nematodes existed all year round.  相似文献   

17.
The occurrence of ectoparasites in sheep flocks is frequently reported but seldom quantified. Sheep production used to be a predominantly family activity in the state of Sa~o Paulo (Brazil), but it began to become a commercial activity in the past decade. Thus, information about the ectoparasites existing in sheep flocks has become necessary. The present data were obtained by means of questionnaires sent to all sheep breeders belonging to the `Associaça~o Paulista de Criadores de Ovinos' (ASPACO; Sa~o Paulo State Association of Sheep Breeders). Response reliability was tested by means of random visits paid to 10.6% of the respondents. Most of the properties (89.5%) reported the presence of one or more ectoparasites. Screw-worm (Cochliomyia hominivorax) was the most frequent ectoparasite (72.5%), followed by bot fly larvae (Dermatobia hominis, 45.0%), ticks (Amblyomma cajennense) and Boophilus microplus, 31.3%) and finally lice (Damalinia ovis, 13.8%). Combined infestations also occurred, the most common one being screw-worm with bot fly larvae (36.0%) followed by bot fly larvae with ticks (13.9%), screw-worm with ticks (9.3%), bot fly larvae with lice (6.9%), and ticks with lice (5.0%). The most common triple combination was screw-worm, bot fly larvae and ticks (12.8%). Breeds raised for meat or wool were attacked by bot fly larvae and ticks more often than other breeds. Lice were only absent from animals of indigenous breeds. The relationships among these ectoparasites are discussed in terms of sheep breeds, flock size, seasonality and the ectoparasitic combinations on the host.  相似文献   

18.
The purpose of this study was to determine the frequency of different tumor types within a large cohort of cats with intracranial neoplasia and to attempt to correlate signalment, tumor size and location, and survival time for each tumor. Medical records of 160 cats with confirmed intracranial neoplasia evaluated between 1985 and 2001 were reviewed. Parameters evaluated included age, sex, breed, FeLV/FIV status, clinical signs, duration of signs, number of tumors, tumor location(s), imaging results, treatment, survival times, and histopathologic diagnosis. Most of the cats were older (11.3 +/- 3.8 years). Primary tumors accounted for 70.6% of cases. Metastasis and direct extension of secondary tumors accounted for only 5.6 and 3.8% of cases, respectively. Twelve cats (7.5%) had 2 or more discrete tumors of the same type, whereas 16 cats (10.0%) had 2 different types of intracranial tumors. The most common tumor types were meningioma (n = 93, 58.1%), lymphoma (n = 23, 14.4%), pituitary tumors (n = 14, 8.8%), and gliomas (n = 12, 7.5%). The most common neurological signs were altered consciousness (n = 42, 26.2%), circling (n = 36, 22.5%), and seizures (n = 36, 22.5%). Cats without specific neurological signs were common (n = 34, 21.2%). The tumor was considered an incidental finding in 30 (18.8%) cats. In addition to expected relationships (eg, meninges and meningioma, pituitary and pituitary tumors), we found that lesion location was predictive of tumor type with diffuse cerebral or brainstem involvement predictive of lymphoma and third ventricle involvement predictive of meningioma.  相似文献   

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20.
An experiment was conducted to study disappearance of C14 to C18 fatty acids, lag times and biohydrogenation (BH) rates of C18 fatty acids of ground soya bean and canola seeds in situ. Three ruminally fistulated Dallagh sheep were used to determine ruminal BH of unsaturated fatty acids (UFAs). Differences in the disappearance of fatty acids through the bags and lag times were observed between the oilseeds. We saw that the longer the incubation time of the oilseeds in the rumen, the lower the content of C18:2 and C18:3. Significantly higher lag times for both C18:2 and C18:3 were observed in ground canola compared to ground soya bean. BH rates of C18:2 and C18:3 fatty acids in soya bean were three times higher than those of canola. These results suggest that the fatty acid profile of fat source can affect the BH of UFAs by rumen micro‐organisms. So that UFAs of canola had higher ability to escape from ruminal BH. It seems that fatty acid profile of ruminant products is more affected by canola seed compared to soya bean seed.  相似文献   

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