采用RACE-PCR方法,从鲢(Hypophthalmichthys molitrix)头肾SMARTcDNA中获得了679 bp IL-8 cDNA序列。结果显示:该序列包含169 bp的5′非编码区,213 bp 3′端非编码区和297 bp的开放阅读框;鲢IL-8的开放阅读框编码98个氨基酸残基,其中包...采用RACE-PCR方法,从鲢(Hypophthalmichthys molitrix)头肾SMARTcDNA中获得了679 bp IL-8 cDNA序列。结果显示:该序列包含169 bp的5′非编码区,213 bp 3′端非编码区和297 bp的开放阅读框;鲢IL-8的开放阅读框编码98个氨基酸残基,其中包含构成两对二硫键的4个保守半胱氨酸。RT-PCR结果显示鲢:IL-8 mRNA主要在胰、肝脏和头肾中表达。将鲢IL-8不含信号肽的成熟多肽克隆到原核融合表达载体pQE30上,转入大肠杆菌M15内进行表达,经SDS-PAGE电泳后显示,重组融合蛋白在分子量约11kD处有一明显表达带,与预期计算的分子量大小相一致。展开更多
肿瘤坏死因子（TNF）是一种炎症细胞因子，在非特异性免疫系统中发挥着重要作用。TNF由单核细胞或巨噬细胞产生，能直接造成肿瘤细胞的死亡，并参与机体炎症和免疫应答的调节。用RACE（rapid amplification of cDNA ends）-PCR方法，从...肿瘤坏死因子（TNF）是一种炎症细胞因子，在非特异性免疫系统中发挥着重要作用。TNF由单核细胞或巨噬细胞产生，能直接造成肿瘤细胞的死亡，并参与机体炎症和免疫应答的调节。用RACE（rapid amplification of cDNA ends）-PCR方法，从鲢总RNA反转录产物中获得了1254 bp TNF cDNA全序列。该序列包含394bp的5’端非编码区，398 bp的3’端非编码区和462bp的开放阅读框。鲢TNF的开放阅读框编码239个氨基酸，其中包含构成一对二硫键的2个保守半胱氨酸。同时利用RT-PCR技术，对该基因在鲢鱼体内不同组织之间的表达差异进行了分析研究，结果表明鲢TNF mRNA主要在脑、鳃和头肾中表达，肠和脾中有少量表达，而在肝中几乎没有表达。展开更多
Objective : Effects of the decoction of Coptis chinensis Franch. , Scutellaria baicalensis Georgi , P. chinense schneid and Lonicera japonica Thunb. on burn rats with wound infection were observed. Methods : Fifty-six...Objective : Effects of the decoction of Coptis chinensis Franch. , Scutellaria baicalensis Georgi , P. chinense schneid and Lonicera japonica Thunb. on burn rats with wound infection were observed. Methods : Fifty-six rats were randomly divided into three groups :sham burn , burn wound infected control(BWIC) and burn wound infected and treatment(BWIT). The plasma was harvested from the experimental rats. Plasma IL-6 activity was measured by using the IL-6-dependent cell line B9. The isolated mono-nuclear cells (MNCs) were induced by LPS and cultured for 24 h. TNF in the culture supernatant was detected by ELISA. Results: Plasma IL-6 levels of BWIC and BWIT were significantly elevated at postburn (PB) 24 and 48h , and IL-6 level of BWIC was significantly higher than that of BWIT at PB 48h (P<0. 01). The induced TNF level of BWIT was significantly higher than that of BWIC at PB 24 and 48 h , and that of sham burn at 24 h(P<0. 01). Conclusion : The results indicate that this decoction could inhibit MNCs releasing from pro-inflammatory cytokines which are believed as an endogenous pyrogen.展开更多
文摘采用RACE-PCR方法,从鲢(Hypophthalmichthys molitrix)头肾SMARTcDNA中获得了679 bp IL-8 cDNA序列。结果显示:该序列包含169 bp的5′非编码区,213 bp 3′端非编码区和297 bp的开放阅读框;鲢IL-8的开放阅读框编码98个氨基酸残基,其中包含构成两对二硫键的4个保守半胱氨酸。RT-PCR结果显示鲢:IL-8 mRNA主要在胰、肝脏和头肾中表达。将鲢IL-8不含信号肽的成熟多肽克隆到原核融合表达载体pQE30上,转入大肠杆菌M15内进行表达,经SDS-PAGE电泳后显示,重组融合蛋白在分子量约11kD处有一明显表达带,与预期计算的分子量大小相一致。
文摘肿瘤坏死因子（TNF）是一种炎症细胞因子，在非特异性免疫系统中发挥着重要作用。TNF由单核细胞或巨噬细胞产生，能直接造成肿瘤细胞的死亡，并参与机体炎症和免疫应答的调节。用RACE（rapid amplification of cDNA ends）-PCR方法，从鲢总RNA反转录产物中获得了1254 bp TNF cDNA全序列。该序列包含394bp的5’端非编码区，398 bp的3’端非编码区和462bp的开放阅读框。鲢TNF的开放阅读框编码239个氨基酸，其中包含构成一对二硫键的2个保守半胱氨酸。同时利用RT-PCR技术，对该基因在鲢鱼体内不同组织之间的表达差异进行了分析研究，结果表明鲢TNF mRNA主要在脑、鳃和头肾中表达，肠和脾中有少量表达，而在肝中几乎没有表达。
文摘Objective : Effects of the decoction of Coptis chinensis Franch. , Scutellaria baicalensis Georgi , P. chinense schneid and Lonicera japonica Thunb. on burn rats with wound infection were observed. Methods : Fifty-six rats were randomly divided into three groups :sham burn , burn wound infected control(BWIC) and burn wound infected and treatment(BWIT). The plasma was harvested from the experimental rats. Plasma IL-6 activity was measured by using the IL-6-dependent cell line B9. The isolated mono-nuclear cells (MNCs) were induced by LPS and cultured for 24 h. TNF in the culture supernatant was detected by ELISA. Results: Plasma IL-6 levels of BWIC and BWIT were significantly elevated at postburn (PB) 24 and 48h , and IL-6 level of BWIC was significantly higher than that of BWIT at PB 48h (P<0. 01). The induced TNF level of BWIT was significantly higher than that of BWIC at PB 24 and 48 h , and that of sham burn at 24 h(P<0. 01). Conclusion : The results indicate that this decoction could inhibit MNCs releasing from pro-inflammatory cytokines which are believed as an endogenous pyrogen.
