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环形泰勒虫喀什株感染牛淋巴细胞系的建立及其生物学特征的鉴定 被引量:2
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作者 马全英 李有全 +8 位作者 刘军龙 刘爱红 王锦明 赵帅阳 陈银银 徐建林 殷宏 关贵全 罗建勋 《中国兽医科学》 CAS CSCD 北大核心 2019年第2期144-151,共8页
建立1株环形泰勒虫(Theileria annulata)感染的牛淋巴细胞系,分析其生物学特征,为环形泰勒虫病疫苗研发提供新的细胞系。用环形泰勒虫喀什株感染的小亚璃眼蜱成蜱叮咬健康黄牛,当动物体温升高,体表淋巴结肿大时,用显微镜检查肩前和股前... 建立1株环形泰勒虫(Theileria annulata)感染的牛淋巴细胞系,分析其生物学特征,为环形泰勒虫病疫苗研发提供新的细胞系。用环形泰勒虫喀什株感染的小亚璃眼蜱成蜱叮咬健康黄牛,当动物体温升高,体表淋巴结肿大时,用显微镜检查肩前和股前淋巴结穿刺物涂片,出现裂殖体感染的淋巴细胞时,手术摘取淋巴结,无菌条件下进行淋巴细胞的分离培养;选择合适的培养条件对该细胞进行适应性传代培养,可稳定传代培养后,绘制细胞生长曲线,测定细胞倍增时间和细胞周期,测定细胞系环形泰勒虫18S rRNA基因序列,通过序列比对分析其亲缘关系。结果显示,建立了1株可传代培养的环形泰勒虫喀什株裂殖体感染的淋巴细胞系(TaKashi),接种5.0×10~5 mL^(-1)的细胞,经72 h培养周期,细胞浓度可达到(2.8~3.0)×10~6mL^(-1),细胞存活率维持在90%以上;细胞群体倍增时间为33.4 h。细胞周期分析显示,G1期占46.2%,G2期占10.4%,S1期占43.4%,TaKashi传50代后仍然保持着淋巴细胞的形态学特征。18S rRNA基因序列比对及抗原基因Tams1同源性分析显示,该虫株与我国不同省份流行的环形泰勒虫虫株的相似性在91.9%以上,并处于同一个分支。 展开更多
关键词 环形泰勒虫 细胞培养 牛淋巴细胞 细胞
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感染环形泰勒虫裂殖体的牛淋巴细胞酵母双杂交文库的构建 被引量:1
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作者 岑双庆 李有全 +9 位作者 刘志杰 陈泽 杨吉飞 关贵全 牛庆丽 任巧云 刘爱红 罗建勋 殷宏 岳城 《中国兽医科学》 CAS CSCD 北大核心 2014年第7期667-671,共5页
为研究环形泰勒虫裂殖体与牛淋巴细胞之间的相互作用,以感染了环形泰勒虫裂殖体的淋巴细胞系为材料,构建了感染环形泰勒虫裂殖体的牛淋巴细胞系的酵母双杂交cDNA文库。首先,提取纯化细胞系的mRNA,经反转录合成第一条链cDNA,通过LD-PCR... 为研究环形泰勒虫裂殖体与牛淋巴细胞之间的相互作用,以感染了环形泰勒虫裂殖体的淋巴细胞系为材料,构建了感染环形泰勒虫裂殖体的牛淋巴细胞系的酵母双杂交cDNA文库。首先,提取纯化细胞系的mRNA,经反转录合成第一条链cDNA,通过LD-PCR合成双链cDNA。柱层析剔除250bp以下的小片段;将双链cDNA与经SmaⅠ处理过的文库质粒pGADT7-Rec一起转入到酵母Y187感受态细胞内,双链cDNA与文库质粒会发生同源重组,经过抗性筛选即可得到酵母双杂交cDNA文库。文库构建结果显示,该文库库容量达2.4×107 CFU,插入片段的平均长度在1 000bp左右,达到试剂盒建库要求。 展开更多
关键词 环形泰勒虫 牛淋巴细胞 酵母双杂交 CDNA文库
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奶牛活性花环(E_a)形成条件的探讨
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作者 焦天嗣 秦永珍 +3 位作者 黎立 黄乐民 丁庆伟 潘瑞萍 《八一农学院学报》 1992年第2期28-31,共4页
用常规 E_α花环形成试验检测牛的花环率,平均数为4.10%.用AET 处理绵羊红细胞后花环率升高,平均数为11.34%。再用 PHA 处理牛淋巴细胞后花环率更高,平均数为52.59%。形成的花环多为球形,稳定、重复性好.
关键词 牛淋巴细胞 花环率 AEI:PHA
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Lymphocytes in patients with psoriasis promote proliferation of ker-atinocytes 被引量:1
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作者 邓安梅 仲人前 +2 位作者 陈孙孝 周晔 孔宪涛 《Journal of Medical Colleges of PLA(China)》 CAS 2002年第1期51-53,共3页
Objective: To analyze the effect of lymphocytes on proliferation of keratinocytes in patients with psoriasis. Methods: Lymphocytes in lesion and peripheral blood were isolated and amplified, then cultured together wit... Objective: To analyze the effect of lymphocytes on proliferation of keratinocytes in patients with psoriasis. Methods: Lymphocytes in lesion and peripheral blood were isolated and amplified, then cultured together with normal keratinocytes. By MTT method, the living cells were quantified in the mixed culture. Results: Compared with normal controls, lymphocytes from lesion and peripheral blood of psoriasis both promote the proliferation of keratinocytes (P<0. 01 and P<0. 05 respectively). The concentrations of IL-2 and IFN-7 in the mixture of lesion lymphocytes and keratinocytes were significantly higher than that of controls. Tripterygium glycosides inhibited this promotion. Conclusion: Lymphocytes in patients with psoriasis (mainly Th1 cell) play an important role in proliferation of keratinocytes. This psoriasis cell model is useful for studies on signal transduction in psoriasis. 展开更多
关键词 PSORIASIS LYMPHOCYTE KERATINOCYTE
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Effect of Metallothionein on Cell Cycle,Apoptosis Rate and Subsets Distribution of Lymphocytes in Peripheral Blood of Dairy Cattle under Heat Stress 被引量:1
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作者 Jiajie LUO Lili LI +7 位作者 Bin ZHANG LiZhuan WU Zijun LI Ying PENG JueXin FAN XinYi LAN JinShun ZHAN Jun FANG 《Agricultural Science & Technology》 CAS 2013年第10期1457-1461,共5页
[Objective] This study aimed to research the effect of metallothionein on cell cycle, apoptosis rate and subsets distribution of lymphocytes in peripheral blood of dairy cows under heat stress, so as to perfect the re... [Objective] This study aimed to research the effect of metallothionein on cell cycle, apoptosis rate and subsets distribution of lymphocytes in peripheral blood of dairy cows under heat stress, so as to perfect the regulative mechanism re- searches of MT to anti-heat stress. [Method] Twenty lactating Chinese Holstein cows were randomly divided into four groups (A, B, C and D), and injected with 0, 4.0, 8.0 and 12.0 mg Zn-metallothionein, respectively by intravenous route. Blood sam- ples were collected at 1", 16~, 31~, 46~ and 61~ day, and the dynamic changes of cell cycle, apoptosis rate and subsets distribution of lymphocytes were determined. [Result] The apoptosis rate of cells in group B and C was lower than those in group A by 26.63% (P〉0.05) and 24.84% (P〉0.05) respectively. The number of cells in the Gc/G1 phage in trial groups was increased and the number of cells in the S and GJM phages tended to decrease, but there were no significant differences (P〉 0.05). The number of CD3~ T cell in three trial groups was greater than those in group A by 7.02% (P〉0.05), 5.45% (P〉0.05) and 3.85% (P〉0.05) respectively, while the number of CD4~ T cell in trial groups was higher than those in control group by 31.04% (P〈0.05), 35.68% (P〈0.05) and 39.34% (P〈0.05) respectively. The number of CD8' T cell and the levels of CD4*/CD8~ in trial groups were increased observ- ably, but significant difference (P〈0.05) was observed in the levels of CD4~/CD8~ between groups A and C only. It demonstrated that exogenous Zn-metallothionein can decrease apoptosis rate, improve cell cycle and regulate subsets distribution of lymphocytes in dairy cattle in a dose-dependent manner. [Conclusion] This study will provide scientific basis for safe utilization of MT in dairy industry. 展开更多
关键词 Exogenous metallothionein Heat stress Immunity Dynamic change Chinese Holstein cows
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Effects of appendectomy and oral tolerance on dextran sulfate sodium colitis 被引量:1
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作者 Min Yue Zhe Shen Chao-Hui Yu Hua Ye Yue-Fang Ye You-Ming Li 《World Journal of Gastroenterology》 SCIE CAS CSCD 2011年第19期2437-2445,共9页
AIM: To evaluate the concomitant effects of appendec- tomy and oral tolerance on colitis.METHODS: Delayed-type hypersensitivity (DTH) was investigated at a 7-d interval after ovalbumin (OVA) ad- ministration and... AIM: To evaluate the concomitant effects of appendec- tomy and oral tolerance on colitis.METHODS: Delayed-type hypersensitivity (DTH) was investigated at a 7-d interval after ovalbumin (OVA) ad- ministration and immunization under normal and colitis conditions in appendectomized or sham-operated mice. Pathological scores for the colon were graded after in- gestion of colon-extracted protein (CEP) and induction of dextran sulfate sodium (DSS) colitis in appendecto- mized or sham-operated mice. Thereafter, Thl and Th2 in Peyer's patches and spleen lymphocytes were de- tected in CEP-treated and bovine serum albumin (BSA)-treated control mice.RESULTS: In appendectomized mice, DTH was not inhibited at day 7 after OVA administration and at the initial phase of DSS colitis, whereas it was inhibited at day 14 and day 21. However, in sham-operated mice, it was inhibited during the whole procedure and the onset of DSS colitis. The protective role of CEP against DSScolitis was present in sham-operated mice, with pre- dominant improvement of colonic pathological changes, while vanished in the appendectomized mice. A shift from Thl to Th2 in Peyer's patches resulted from a de- crease of Thl cells with the ingestion of CEP. Compared with BSA in the sham-operated group, no predominant changes were observed in the appendectomized mice.CONCLUSION: Appendectomy interferes with the pro- tective role of CEP in DSS colitis via a shift from Th2 to Thl during oral tolerance induction. 展开更多
关键词 APPENDICECTOMY Oral tolerance Dextrin sulfate sodium Murine colitis Th1-Th2 balance
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