为建立一种能快速对羊泰勒虫(ovine and caprine theileria)和嗜吞噬细胞无浆体(A.phagocytoph-ilum)同时进行检测的双重PCR方法。根据GenBank已报道的羊泰勒虫MPSP基因和羊嗜吞噬细胞无浆体16S rRNA基因设计合成了2对特异性引物,通过...为建立一种能快速对羊泰勒虫(ovine and caprine theileria)和嗜吞噬细胞无浆体(A.phagocytoph-ilum)同时进行检测的双重PCR方法。根据GenBank已报道的羊泰勒虫MPSP基因和羊嗜吞噬细胞无浆体16S rRNA基因设计合成了2对特异性引物,通过条件优化,建立了双重PCR检测方法。双重PCR可特异扩增出羊泰勒虫和嗜吞噬细胞无浆体目的条带,片段大小分别为875bp和394bp。该方法具有较好的特异性,对羊泰勒虫和嗜吞噬细胞无浆体的最低检出浓度为16fg/μL和1fg/μL。对采集的60份血液样本进行双重PCR检测,羊泰勒虫阳性率为46.67%(28/60),嗜吞噬细胞无浆体阳性率为13.33%(8/60),混合感染率为10%(6/60)。结果表明,双重PCR方法可用于羊泰勒虫和嗜吞噬细胞无浆体的快速诊断和流行病学调查。展开更多
OBJECTIVE: To investigate the hormone-like activities of Kuntai capsule(KTC) in the uteri of ovariectomized rats and immature rabbits.METHODS: Following bilateral ovariectomy, rats were randomly divided into six group...OBJECTIVE: To investigate the hormone-like activities of Kuntai capsule(KTC) in the uteri of ovariectomized rats and immature rabbits.METHODS: Following bilateral ovariectomy, rats were randomly divided into six groups including sham group, control group, estradiol valerate group, KTC 0.24, 0.6, and 1.5 g/kg groups. The rats were treated with 0.5% CMC-Na, estradiol valerate and KTC(0.24, 0.6, and 1.5 g/kg), respectively for 28 consecutive days. Then the estrous cycle, uterine changes and pathological changes were examined.Serum levels of estradiol(E2), and progesterone(P4)were measured by enzyme-linked immunosorbent assay(ELISA). Protein levels of estradiol receptor(ER), progesterone receptor(PR), vascular endothelial growth factor(VEGF), proliferating cell nuclear antigen(PCNA) and nuclear-associated antigen-67(Ki-67) in uterine tissues were detected by western blot. Immature rabbits were estrogen-primed prior to intragastric administration with KTC for 6 d consecutively. Then, the uteri underwent hematoxylin-eosin staining to observe endometrial transformation.RESULTS: Compared with the control group, 0.6and 1.5 g/kg KTC markedly decreased the uterine organ coefficient and endometrial thickness(P <0.05). The serum level of P4 was increased in the KTC 0.6 g/kg group(P < 0.05). There were no significant variations in the serum level of E2 in the KTC groups compared with the control group. ERβ, but not ERα, was markedly upregulated after KTC administration(P < 0.05). Furthermore, 1.5 g/kg KTC significantly decreased the protein level of PRA(P <0.05) and 0.6 g/kg KTC increased the protein level of PRB in the uteri(P < 0.05). VEGF was highly expressed after treatment with 0.24 and 0.6 g/kg KTC,and Ki-67 was markedly reduced in ovariectomized rats treated with 1.5 g/kg KTC. No difference was found in the expression of PCNA. KTC 0.24 and0.6 g/kg promoted endometrial transformation in immature rabbit uteri.CONCLUSION: KTC does not demonstrate obvious estrogen-like effect on uteri after ovariectomy, but it does exhibit weak progestogen-like effect, by which mechanism of action is yet to be further investigated.展开更多
文摘为建立一种能快速对羊泰勒虫(ovine and caprine theileria)和嗜吞噬细胞无浆体(A.phagocytoph-ilum)同时进行检测的双重PCR方法。根据GenBank已报道的羊泰勒虫MPSP基因和羊嗜吞噬细胞无浆体16S rRNA基因设计合成了2对特异性引物,通过条件优化,建立了双重PCR检测方法。双重PCR可特异扩增出羊泰勒虫和嗜吞噬细胞无浆体目的条带,片段大小分别为875bp和394bp。该方法具有较好的特异性,对羊泰勒虫和嗜吞噬细胞无浆体的最低检出浓度为16fg/μL和1fg/μL。对采集的60份血液样本进行双重PCR检测,羊泰勒虫阳性率为46.67%(28/60),嗜吞噬细胞无浆体阳性率为13.33%(8/60),混合感染率为10%(6/60)。结果表明,双重PCR方法可用于羊泰勒虫和嗜吞噬细胞无浆体的快速诊断和流行病学调查。
文摘OBJECTIVE: To investigate the hormone-like activities of Kuntai capsule(KTC) in the uteri of ovariectomized rats and immature rabbits.METHODS: Following bilateral ovariectomy, rats were randomly divided into six groups including sham group, control group, estradiol valerate group, KTC 0.24, 0.6, and 1.5 g/kg groups. The rats were treated with 0.5% CMC-Na, estradiol valerate and KTC(0.24, 0.6, and 1.5 g/kg), respectively for 28 consecutive days. Then the estrous cycle, uterine changes and pathological changes were examined.Serum levels of estradiol(E2), and progesterone(P4)were measured by enzyme-linked immunosorbent assay(ELISA). Protein levels of estradiol receptor(ER), progesterone receptor(PR), vascular endothelial growth factor(VEGF), proliferating cell nuclear antigen(PCNA) and nuclear-associated antigen-67(Ki-67) in uterine tissues were detected by western blot. Immature rabbits were estrogen-primed prior to intragastric administration with KTC for 6 d consecutively. Then, the uteri underwent hematoxylin-eosin staining to observe endometrial transformation.RESULTS: Compared with the control group, 0.6and 1.5 g/kg KTC markedly decreased the uterine organ coefficient and endometrial thickness(P <0.05). The serum level of P4 was increased in the KTC 0.6 g/kg group(P < 0.05). There were no significant variations in the serum level of E2 in the KTC groups compared with the control group. ERβ, but not ERα, was markedly upregulated after KTC administration(P < 0.05). Furthermore, 1.5 g/kg KTC significantly decreased the protein level of PRA(P <0.05) and 0.6 g/kg KTC increased the protein level of PRB in the uteri(P < 0.05). VEGF was highly expressed after treatment with 0.24 and 0.6 g/kg KTC,and Ki-67 was markedly reduced in ovariectomized rats treated with 1.5 g/kg KTC. No difference was found in the expression of PCNA. KTC 0.24 and0.6 g/kg promoted endometrial transformation in immature rabbit uteri.CONCLUSION: KTC does not demonstrate obvious estrogen-like effect on uteri after ovariectomy, but it does exhibit weak progestogen-like effect, by which mechanism of action is yet to be further investigated.