[目的]探讨血清磷脂酶A2组ⅡA (phospholipase A2 groupⅡA,PLA2G2A)和凝血因子Ⅸ(coagulation factorⅨ,F9)对耐多药肺结核病(multidrug-resistant tuberculosis,MDR-TB)气阴两虚(deficiency of Qi and Yin,DQY)证的潜在诊断价值。[方...[目的]探讨血清磷脂酶A2组ⅡA (phospholipase A2 groupⅡA,PLA2G2A)和凝血因子Ⅸ(coagulation factorⅨ,F9)对耐多药肺结核病(multidrug-resistant tuberculosis,MDR-TB)气阴两虚(deficiency of Qi and Yin,DQY)证的潜在诊断价值。[方法]选择MDR-TB肺阴虚(pulmonary Yin deficiency,PYD)证、MDR-TB阴虚火旺(hyperactivity of fire due to Yin deficiency,HFYD)证、MDR-TB DQY证患者和健康对照(healthy control,HC)者各15例,以ELISA检测每组PLA2G2A和F9的蛋白浓度,并利用受试者工作特征(receiver operating characteristic, ROC)曲线分析PLA2G2A和F9作为MDR-TB DQY证潜在生物学标志物的临床价值。[结果]MDR-TB证候分布以DQY证患者数最多,其次是HFYD证,再者是DQY证,而阴阳两虚(deficiency of Yin and Yang, DYY)证患者数最少。DQY证组PLA2G2A的蛋白浓度均明显高于PYD证组、HFYD证组和HC组,差异有统计学意义(P<0.05);DQY证组F9的蛋白浓度明显低于HFYD证组,差异有统计学意义(P<0.05)。ROC曲线结果显示,血清PLA2G2A有鉴别诊断MDR-TB DQY证的作用。[结论]血清PLA2G2A和F9在诊断MDR-TB DQY证方面具备一定临床价值,血清PLA2G2A可作为MDR-TB DQY证的潜在生物学标志物。展开更多
Human FⅨ expression vector pCMVⅨ was packaged by EffecteneTM reagent and injected into mice seminiferous tubules with glass pipettes. The expressional frame of pCMVⅨ was examined by PCR and Southern blotting among ...Human FⅨ expression vector pCMVⅨ was packaged by EffecteneTM reagent and injected into mice seminiferous tubules with glass pipettes. The expressional frame of pCMVⅨ was examined by PCR and Southern blotting among 41 progenies. There were 2 (4%) mice being integrated with hFⅨ gene into chromosomes. 4.6 ng/mL of hFⅨ protein was expressed in plasma of one mouse, which was tested by ELISA. We demonstrated that building of transgenic animals by spermatogonial stem cells is an efficient method. Meanwhile, it has also been proved to be an alternative choice for mammary gland bioreactor.展开更多
The recombinant adeno-associated viral vector with human coagulation Factor Ⅸ minigene which wasregulated by CMV promoter was constructed. Largequantity of recombinant adeno-associated viral particles (rAAV/ hFⅨ) wa...The recombinant adeno-associated viral vector with human coagulation Factor Ⅸ minigene which wasregulated by CMV promoter was constructed. Largequantity of recombinant adeno-associated viral particles (rAAV/ hFⅨ) was prepared by the HSV/AAV hybrid helper virus method. Southern dot blot assay and QC-PCR indicated that the titer of the virus was 3.6×1012 v.g./mL. It demonstrated that this method can effectively overcome the hurdles of mass production of AAV vector. Followed by anintramuscular injection of viral vectors (7.5×1011 v.g./mouse) in the quadriceps femoris, an elevation of human Factor Ⅸexpression in the plasma of hemophilia B mice was detected (387 ng/mL) and persisted more than 12 weeks. The level of anti-virus antibody in plasma aligned with the Factor Ⅸexpression curve. The QC-PCR method is easier and moreaccurate than traditional dot hybridization fordetermination of the titer of recombinant adeno-associated virus. Moreover, there are no HSV particles existing inproduced AAV assayed by RT-PCR. AAV is the only virus that has been amplified from AAV-injected muscle by PCR.展开更多
文摘[目的]探讨血清磷脂酶A2组ⅡA (phospholipase A2 groupⅡA,PLA2G2A)和凝血因子Ⅸ(coagulation factorⅨ,F9)对耐多药肺结核病(multidrug-resistant tuberculosis,MDR-TB)气阴两虚(deficiency of Qi and Yin,DQY)证的潜在诊断价值。[方法]选择MDR-TB肺阴虚(pulmonary Yin deficiency,PYD)证、MDR-TB阴虚火旺(hyperactivity of fire due to Yin deficiency,HFYD)证、MDR-TB DQY证患者和健康对照(healthy control,HC)者各15例,以ELISA检测每组PLA2G2A和F9的蛋白浓度,并利用受试者工作特征(receiver operating characteristic, ROC)曲线分析PLA2G2A和F9作为MDR-TB DQY证潜在生物学标志物的临床价值。[结果]MDR-TB证候分布以DQY证患者数最多,其次是HFYD证,再者是DQY证,而阴阳两虚(deficiency of Yin and Yang, DYY)证患者数最少。DQY证组PLA2G2A的蛋白浓度均明显高于PYD证组、HFYD证组和HC组,差异有统计学意义(P<0.05);DQY证组F9的蛋白浓度明显低于HFYD证组,差异有统计学意义(P<0.05)。ROC曲线结果显示,血清PLA2G2A有鉴别诊断MDR-TB DQY证的作用。[结论]血清PLA2G2A和F9在诊断MDR-TB DQY证方面具备一定临床价值,血清PLA2G2A可作为MDR-TB DQY证的潜在生物学标志物。
文摘Human FⅨ expression vector pCMVⅨ was packaged by EffecteneTM reagent and injected into mice seminiferous tubules with glass pipettes. The expressional frame of pCMVⅨ was examined by PCR and Southern blotting among 41 progenies. There were 2 (4%) mice being integrated with hFⅨ gene into chromosomes. 4.6 ng/mL of hFⅨ protein was expressed in plasma of one mouse, which was tested by ELISA. We demonstrated that building of transgenic animals by spermatogonial stem cells is an efficient method. Meanwhile, it has also been proved to be an alternative choice for mammary gland bioreactor.
基金supported by the National Natural Science Foundation of China(Grant No.30100102).
文摘The recombinant adeno-associated viral vector with human coagulation Factor Ⅸ minigene which wasregulated by CMV promoter was constructed. Largequantity of recombinant adeno-associated viral particles (rAAV/ hFⅨ) was prepared by the HSV/AAV hybrid helper virus method. Southern dot blot assay and QC-PCR indicated that the titer of the virus was 3.6×1012 v.g./mL. It demonstrated that this method can effectively overcome the hurdles of mass production of AAV vector. Followed by anintramuscular injection of viral vectors (7.5×1011 v.g./mouse) in the quadriceps femoris, an elevation of human Factor Ⅸexpression in the plasma of hemophilia B mice was detected (387 ng/mL) and persisted more than 12 weeks. The level of anti-virus antibody in plasma aligned with the Factor Ⅸexpression curve. The QC-PCR method is easier and moreaccurate than traditional dot hybridization fordetermination of the titer of recombinant adeno-associated virus. Moreover, there are no HSV particles existing inproduced AAV assayed by RT-PCR. AAV is the only virus that has been amplified from AAV-injected muscle by PCR.
