AIM: The goal of this study was to characterize the AFP receptor, its possible signal transduction pathway and its proliferative functions in human hepatoma cell line Bel 7402. METHODS: Cell proliferation enhanced by ...AIM: The goal of this study was to characterize the AFP receptor, its possible signal transduction pathway and its proliferative functions in human hepatoma cell line Bel 7402. METHODS: Cell proliferation enhanced by AFP was detected by MTT assay, 3H-thymidine incorporation and S-stage percentage of cell cycle analysis. With radioactive labeled 125I-AFP for receptor binding assay; cAMP accumulation, protein kinase A activity were detected by radioactive immunosorbent assay and the change of intracellular free calcium (Ca2+i) was monitored by scanning fluorescence intensity under TCS-NT confocal microscope. The expression of oncogenes N- ras, p 53, and p21( ras ) in the cultured cells in vitro were detected by Northern blotting and Western blotting respectively. RESULTS: It was demonstrated that AFP enhanced the proliferation of human hepatoma Bel 7402 cell in a dose dependent fashion as shown in MTT assay, (3)H-thymidine incorporation and S-phase percentage up to 2-fold. Two subtypes of AFP receptors were identified in the cells with Kds of 1.3 x 10(-9)mol.L(-1) and 9.9 x10(-8)mol. (-1)L respectively. Pretreatment of cells with AFP resulted in a significant increase (625%) in cAMP accumulation. The activity of protein kinase A activity were increased up to 37.5, 122.6, 73.7 and 61.2% at treatment time point 2, 6, 12 and 24 hours. The level of intracellular calcium were elevated after the treatment of alpha-fetoprotein and achieved to 204% at 4 min. The results also showed that AFP(20mg.L(-1)) could upregulate the expression of N- ras oncogenes and p 53 and p21( ras ) in Bel 7402 cells. In the later case,the alteration were 81.1%(12h) and 97.3%(12h) respectively compared with control. CONCLUSION: These results demonstrate that AFP is a potential growth factor to promote the proliferation of human hepatoma Bel 7402 cells. Its growth-regulatory effects are mediated by its specific plasma membrane receptors coupled with its transmembrane signaling transduction through the pathway of cAMP-PKA and intracellular calcium展开更多
AIM: The existence and properties of alpha-fetoprotein (AFP) receptor on the surface of NIH 3T3 cells and the effects of AFP on cellular signal transduction pathway were investigated. METHODS: The effect of AFP on the...AIM: The existence and properties of alpha-fetoprotein (AFP) receptor on the surface of NIH 3T3 cells and the effects of AFP on cellular signal transduction pathway were investigated. METHODS: The effect of AFP on the proliferation of NIH 3T3 cells was measured by incorporation of 3H-TdR. Receptor-binding assay of 125I-AFP was performed to detect the properties of AFP receptor in NIH 3T3 cells. The influences of AFP on the [cAMP]i and the activities of protein kinase A (PKA) were determined. Western blot was used to detect the change of K-ras P21 protein expression. RESULTS: The proliferation of NIH 3T3 cells treated with 0-80 mg/L of AFP was significantly enhanced. The Scatchard analysis indicated that there were two classes of binding sites with KD of 2.722 x 10(-9)M (Bmax=12810 sites per cell) and 8.931 x 10(-8)M (Bmax=119700 sites per cell) respectively. In the presence of AFP (20 mg/L), the content of cAMP and activities of PKA were significantly elevated . The level of K-ras P21 protein was upregulated by AFP at the concentration of 20 mg/L. The monoclonal antibody against AFP could reverse the effects of AFP on the cAMP content, PKA activity and the expression of K-ras p21 gene. CONCLUSION: The effect of AFP on the cell proliferation was achieved by binding its receptor to trigger the signal transduction pathway of cAMP-PKA and alter the expression of K- ras p21 gene.展开更多
Objective： To observe the curative effect of dissolving phlegm-stasis on knee osteoarthritis. Methods： The 100 patients with knee osteoarthritis were randomly divided into two groups. The 50 patients in the treatmen...Objective： To observe the curative effect of dissolving phlegm-stasis on knee osteoarthritis. Methods： The 100 patients with knee osteoarthritis were randomly divided into two groups. The 50 patients in the treatment group orally took Chinese medicine while the other 50 patients in the control groups orally took Votalin and Vitamin C for one week as a course of treatment. At the end of 2-week treatment, analytic comparison was carried in evaluate the curative effect and the changes in total score of symptoms before and after treatment between the two groups. Results： The total score of symptoms after treatment was significantly lower than that before treatment in both groups （P〈0.01, P〈0.05）. The total score after treatment of patients with X-ray grade I and II in the treatment group was more obviously reduced as compared to the control group （P〈0.05）. The remarkably effective rate after treatment of patients with X-ray grade I and It in the treatment group was also higher than that in the control group （P〈0.05）. Conclusion： The treatment of knee osteoarthritis with dissolving phlegm-stasis can effectively improve the clinical symptoms.展开更多
Summary: In order to study the effects of electromagnetic fields (EMFs) on proliferation, differentiation and intercellular cyclic AMP (cAMP) in mouse bone marrow mesenchymal stem cells (MSCs) in vitro, the mouse bone...Summary: In order to study the effects of electromagnetic fields (EMFs) on proliferation, differentiation and intercellular cyclic AMP (cAMP) in mouse bone marrow mesenchymal stem cells (MSCs) in vitro, the mouse bone MSCs were isolated and cultured in vitro. The third passage MSCs were divided into 4 groups and stimulated with EMFs. The cellular proliferation (MTT), the cellular differentiation (alkaline phosphatase activity, ALP), and the intercellular cAMP level were investigated at different time points. The results showed that EMF (50Hz pulse burst 2 mT peak) inhibited the cellular proliferation (P<0.05), enhanced the cellular differentiation (P<0.05), and increased the intercellular cAMP level (P<0.01) in the early time of the stimulation (1-3 days), but the intercellular cAMP level did not increased further in the later days. We are led to conclude that the cAMP may be involved in the mediation of the growth inhibitory and differentiation-inducing signals of specific EMFs in vitro.展开更多
目的 了解医院感染中高产 Am p Cβ-内酰胺酶阴沟肠杆菌的流行情况。方法 通过药敏实验和等电点分析筛选出高产 Am p Cβ-内酰胺酶的阴沟肠杆菌 ,然后通过诱导实验分析了其 Am p Cβ-内酰胺酶表型 ,最后 ,通过PFGE分型技术考察了其克...目的 了解医院感染中高产 Am p Cβ-内酰胺酶阴沟肠杆菌的流行情况。方法 通过药敏实验和等电点分析筛选出高产 Am p Cβ-内酰胺酶的阴沟肠杆菌 ,然后通过诱导实验分析了其 Am p Cβ-内酰胺酶表型 ,最后 ,通过PFGE分型技术考察了其克隆构成情况。结果 在 35株阴沟肠杆菌中 ,2 8株 (80 % )高产 Am p Cβ-内酰胺酶 ,其中18株 (6 5 % )属完全去抑制型 ,10株 (35 % )属部分去抑制型 ;2 8株高产 Am p Cβ-内酰胺酶阴沟肠杆菌的染色体DNA指纹图呈 17种克隆的多克隆构成模式。结论 阴沟肠杆菌引起的医院感染中流行高产 Amp Cβ-内酰胺酶的菌株 ,完全去抑制型为其主要表型 ;其内源性感染方式使阴沟肠杆菌医院感染株呈多克隆构成模式。展开更多
基金National Natural Science Foundation of China,No.39760077
文摘AIM: The goal of this study was to characterize the AFP receptor, its possible signal transduction pathway and its proliferative functions in human hepatoma cell line Bel 7402. METHODS: Cell proliferation enhanced by AFP was detected by MTT assay, 3H-thymidine incorporation and S-stage percentage of cell cycle analysis. With radioactive labeled 125I-AFP for receptor binding assay; cAMP accumulation, protein kinase A activity were detected by radioactive immunosorbent assay and the change of intracellular free calcium (Ca2+i) was monitored by scanning fluorescence intensity under TCS-NT confocal microscope. The expression of oncogenes N- ras, p 53, and p21( ras ) in the cultured cells in vitro were detected by Northern blotting and Western blotting respectively. RESULTS: It was demonstrated that AFP enhanced the proliferation of human hepatoma Bel 7402 cell in a dose dependent fashion as shown in MTT assay, (3)H-thymidine incorporation and S-phase percentage up to 2-fold. Two subtypes of AFP receptors were identified in the cells with Kds of 1.3 x 10(-9)mol.L(-1) and 9.9 x10(-8)mol. (-1)L respectively. Pretreatment of cells with AFP resulted in a significant increase (625%) in cAMP accumulation. The activity of protein kinase A activity were increased up to 37.5, 122.6, 73.7 and 61.2% at treatment time point 2, 6, 12 and 24 hours. The level of intracellular calcium were elevated after the treatment of alpha-fetoprotein and achieved to 204% at 4 min. The results also showed that AFP(20mg.L(-1)) could upregulate the expression of N- ras oncogenes and p 53 and p21( ras ) in Bel 7402 cells. In the later case,the alteration were 81.1%(12h) and 97.3%(12h) respectively compared with control. CONCLUSION: These results demonstrate that AFP is a potential growth factor to promote the proliferation of human hepatoma Bel 7402 cells. Its growth-regulatory effects are mediated by its specific plasma membrane receptors coupled with its transmembrane signaling transduction through the pathway of cAMP-PKA and intracellular calcium
基金This work was supported by National NaturalScience Fundation of China(No.39760077).
文摘AIM: The existence and properties of alpha-fetoprotein (AFP) receptor on the surface of NIH 3T3 cells and the effects of AFP on cellular signal transduction pathway were investigated. METHODS: The effect of AFP on the proliferation of NIH 3T3 cells was measured by incorporation of 3H-TdR. Receptor-binding assay of 125I-AFP was performed to detect the properties of AFP receptor in NIH 3T3 cells. The influences of AFP on the [cAMP]i and the activities of protein kinase A (PKA) were determined. Western blot was used to detect the change of K-ras P21 protein expression. RESULTS: The proliferation of NIH 3T3 cells treated with 0-80 mg/L of AFP was significantly enhanced. The Scatchard analysis indicated that there were two classes of binding sites with KD of 2.722 x 10(-9)M (Bmax=12810 sites per cell) and 8.931 x 10(-8)M (Bmax=119700 sites per cell) respectively. In the presence of AFP (20 mg/L), the content of cAMP and activities of PKA were significantly elevated . The level of K-ras P21 protein was upregulated by AFP at the concentration of 20 mg/L. The monoclonal antibody against AFP could reverse the effects of AFP on the cAMP content, PKA activity and the expression of K-ras p21 gene. CONCLUSION: The effect of AFP on the cell proliferation was achieved by binding its receptor to trigger the signal transduction pathway of cAMP-PKA and alter the expression of K- ras p21 gene.
文摘Objective： To observe the curative effect of dissolving phlegm-stasis on knee osteoarthritis. Methods： The 100 patients with knee osteoarthritis were randomly divided into two groups. The 50 patients in the treatment group orally took Chinese medicine while the other 50 patients in the control groups orally took Votalin and Vitamin C for one week as a course of treatment. At the end of 2-week treatment, analytic comparison was carried in evaluate the curative effect and the changes in total score of symptoms before and after treatment between the two groups. Results： The total score of symptoms after treatment was significantly lower than that before treatment in both groups （P〈0.01, P〈0.05）. The total score after treatment of patients with X-ray grade I and II in the treatment group was more obviously reduced as compared to the control group （P〈0.05）. The remarkably effective rate after treatment of patients with X-ray grade I and It in the treatment group was also higher than that in the control group （P〈0.05）. Conclusion： The treatment of knee osteoarthritis with dissolving phlegm-stasis can effectively improve the clinical symptoms.
基金This project was supported by a grant from the National Natural Sciences Foundation of China (No. 50347025).
文摘Summary: In order to study the effects of electromagnetic fields (EMFs) on proliferation, differentiation and intercellular cyclic AMP (cAMP) in mouse bone marrow mesenchymal stem cells (MSCs) in vitro, the mouse bone MSCs were isolated and cultured in vitro. The third passage MSCs were divided into 4 groups and stimulated with EMFs. The cellular proliferation (MTT), the cellular differentiation (alkaline phosphatase activity, ALP), and the intercellular cAMP level were investigated at different time points. The results showed that EMF (50Hz pulse burst 2 mT peak) inhibited the cellular proliferation (P<0.05), enhanced the cellular differentiation (P<0.05), and increased the intercellular cAMP level (P<0.01) in the early time of the stimulation (1-3 days), but the intercellular cAMP level did not increased further in the later days. We are led to conclude that the cAMP may be involved in the mediation of the growth inhibitory and differentiation-inducing signals of specific EMFs in vitro.
文摘目的 了解医院感染中高产 Am p Cβ-内酰胺酶阴沟肠杆菌的流行情况。方法 通过药敏实验和等电点分析筛选出高产 Am p Cβ-内酰胺酶的阴沟肠杆菌 ,然后通过诱导实验分析了其 Am p Cβ-内酰胺酶表型 ,最后 ,通过PFGE分型技术考察了其克隆构成情况。结果 在 35株阴沟肠杆菌中 ,2 8株 (80 % )高产 Am p Cβ-内酰胺酶 ,其中18株 (6 5 % )属完全去抑制型 ,10株 (35 % )属部分去抑制型 ;2 8株高产 Am p Cβ-内酰胺酶阴沟肠杆菌的染色体DNA指纹图呈 17种克隆的多克隆构成模式。结论 阴沟肠杆菌引起的医院感染中流行高产 Amp Cβ-内酰胺酶的菌株 ,完全去抑制型为其主要表型 ;其内源性感染方式使阴沟肠杆菌医院感染株呈多克隆构成模式。
