The hormone defined serum free conditioned medium (SFCM) of human nasopharyngeal carcinoma epithelioid cell line (CNE1) was assayed by both the 3H-thymidine incorporation test and the soft agar test. It was found that...The hormone defined serum free conditioned medium (SFCM) of human nasopharyngeal carcinoma epithelioid cell line (CNE1) was assayed by both the 3H-thymidine incorporation test and the soft agar test. It was found that the SFCM stimulated the growth of long-term serum-free cultured CNE4 cells in ac-cordence with the fact that the growth rate of long-term serum-free cultured CNE1 cells was directly proportional to the plating density. Alternatively 5% SFCM inhibited the growth of short-term serum-free cultured CNE4 cells by 51% in which the indicator cell remained the responsiveness state of growing in the serum-supplemented medium to the effector of interest. Furthermore, SFCM resulted in the inhibition of anchorage-independent growth of CNE4 cells and A431 cells. Also in soft agar test. SFCM reduced the colony formation of NRK(?),9F cells in the presence of EGF or EGF plus TGF-β. These finding suggested that CNE4 secreted autocrine growth stimulating factor(s) and growth inhibiting factor(s) in the serum-free medium, the latter strongly reverse malignant phenotypes of CNE4 and A431 cells in serum-supplemented surrounding.展开更多
为提高航空运输集装托盘使用的安全性与经济性,给航空运输集装托盘结构健康监测技术研究提供一种损伤模拟方法,使用幅值和相位综合损伤因子、幅值无关损伤因子和相位无关损伤因子3种损伤因子计算方法,分别计算不同激励信号中心频率下,...为提高航空运输集装托盘使用的安全性与经济性,给航空运输集装托盘结构健康监测技术研究提供一种损伤模拟方法,使用幅值和相位综合损伤因子、幅值无关损伤因子和相位无关损伤因子3种损伤因子计算方法,分别计算不同激励信号中心频率下,粘贴吸波胶、粘贴钢球、结构状态微变和噪声,以及不同面积结构真实损伤的损伤因子。实验结果表明:可以使用粘贴钢球的方式模拟航空运输集装托盘发生损伤,Lamb波的激励信号中心频率宜选择在100~120 k Hz。展开更多
Objective:To investigate the effects of astragalosideⅣ(AS-Ⅳ)on podocyte injury of diabetic nephropathy(DN)and reveal its potential mechanism.Methods:In in vitro experiment,podocytes were divided into 4 groups,normal...Objective:To investigate the effects of astragalosideⅣ(AS-Ⅳ)on podocyte injury of diabetic nephropathy(DN)and reveal its potential mechanism.Methods:In in vitro experiment,podocytes were divided into 4 groups,normal,high glucose(HG),inositol-requiring enzyme 1(IRE-1)αactivator(HG+thapsigargin1μmol/L),and IRE-1αinhibitor(HG+STF-083010,20μmol/L)groups.Additionally,podocytes were divided into 4 groups,including normal,HG,AS-Ⅳ(HG+AS-Ⅳ20μmol/L),and IRE-1αinhibitor(HG+STF-083010,20μmol/L)groups,respectively.After 24 h treatment,the morphology of podocytes and endoplasmic reticulum(ER)was observed by electron microscopy.The expressions of glucose-regulated protein 78(GRP78)and IRE-1αwere detected by cellular immunofluorescence.In in vivo experiment,DN rat model was established via a consecutive 3-day intraperitoneal streptozotocin(STZ)injections.A total of 40 rats were assigned into the normal,DN,AS-Ⅳ[AS-Ⅳ40 mg/(kg·d)],and IRE-1αinhibitor[STF-083010,10 mg/(kg·d)]groups(n=10),respectively.The general condition,24-h urine volume,random blood glucose,urinary protein excretion rate(UAER),blood urea nitrogen(BUN),and serum creatinine(SCr)levels of rats were measured after 8 weeks of intervention.Pathological changes in the renal tissue were observed by hematoxylin and eosin(HE)staining.Quantitative reverse transcription-polymerase chain reaction(RT-PCR)and Western blot were used to detect the expressions of GRP78,IRE-1α,nuclear factor kappa Bp65(NF-κBp65),interleukin(IL)-1β,NLR family pyrin domain containing 3(NLRP3),caspase-1,gasdermin D-N(GSDMD-N),and nephrin at the mRNA and protein levels in vivo and in vitro,respectively.Results:Cytoplasmic vacuolation and ER swelling were observed in the HG and IRE-1αactivator groups.Podocyte morphology and ER expansion were improved in AS-Ⅳand IRE-1αinhibitor groups compared with HG group.Cellular immunofluorescence showed that compared with the normal group,the fluorescence intensity of GRP78 and IRE-1αin the HG and IRE-1αactivator groups were significantly increased whereas decreased in AS-Ⅳand IRE-1αinhibitor groups(P<0.05).Compared with the normal group,the mRNA and protein expressions of GRP78,IRE-1α,NF-κBp65,IL-1β,NLRP3,caspase-1 and GSDMD-N in the HG group was increased(P<0.05).Compared with HG group,the expression of above indices was decreased in the AS-Ⅳand IRE-1αinhibitor groups,and the expression in the IRE-1αactivator group was increased(P<0.05).The expression of nephrin was decreased in the HG group,and increased in AS-Ⅳand IRE-1αinhibitor groups(P<0.05).The in vivo experiment results revealed that compared to the normal group,the levelse of blood glucose,triglyceride,total cholesterol,BUN,SCr and urinary protein in the DN group were higher(P<0.05).Compared with DN group,the above indices in AS-Ⅳand IRE-1αinhibitor groups were decreased(P<0.05).HE staining revealed glomerular hypertrophy,mesangial widening and mesangial cell proliferation in the renal tissue of the DN group.Compared with the DN group,the above pathological changes in renal tissue of AS-Ⅳand IRE-1αinhibitor groups were alleviated.Quantitative RT-PCR and Western blot results of GRP78,IRE-1α,NF-κBp65,IL-1β,NLRP3,caspase-1 and GSDMD-N were consistent with immunofluorescence analysis.Conclusion:AS-Ⅳcould reduce ERS and inflammation,improve podocyte pyroptosis,thus exerting a podocyteprotective effect in DN,through regulating IRE-1α/NF-κB/NLRP3 signaling pathway.展开更多
文摘The hormone defined serum free conditioned medium (SFCM) of human nasopharyngeal carcinoma epithelioid cell line (CNE1) was assayed by both the 3H-thymidine incorporation test and the soft agar test. It was found that the SFCM stimulated the growth of long-term serum-free cultured CNE4 cells in ac-cordence with the fact that the growth rate of long-term serum-free cultured CNE1 cells was directly proportional to the plating density. Alternatively 5% SFCM inhibited the growth of short-term serum-free cultured CNE4 cells by 51% in which the indicator cell remained the responsiveness state of growing in the serum-supplemented medium to the effector of interest. Furthermore, SFCM resulted in the inhibition of anchorage-independent growth of CNE4 cells and A431 cells. Also in soft agar test. SFCM reduced the colony formation of NRK(?),9F cells in the presence of EGF or EGF plus TGF-β. These finding suggested that CNE4 secreted autocrine growth stimulating factor(s) and growth inhibiting factor(s) in the serum-free medium, the latter strongly reverse malignant phenotypes of CNE4 and A431 cells in serum-supplemented surrounding.
文摘为提高航空运输集装托盘使用的安全性与经济性,给航空运输集装托盘结构健康监测技术研究提供一种损伤模拟方法,使用幅值和相位综合损伤因子、幅值无关损伤因子和相位无关损伤因子3种损伤因子计算方法,分别计算不同激励信号中心频率下,粘贴吸波胶、粘贴钢球、结构状态微变和噪声,以及不同面积结构真实损伤的损伤因子。实验结果表明:可以使用粘贴钢球的方式模拟航空运输集装托盘发生损伤,Lamb波的激励信号中心频率宜选择在100~120 k Hz。
基金Supported by the International Cooperation Project of Department of Science and Technology of Shanxi Province(No.201903D421057)the Youth Fund Project of Department of Science and Technology of Shanxi Province(No.201901D211485)。
文摘Objective:To investigate the effects of astragalosideⅣ(AS-Ⅳ)on podocyte injury of diabetic nephropathy(DN)and reveal its potential mechanism.Methods:In in vitro experiment,podocytes were divided into 4 groups,normal,high glucose(HG),inositol-requiring enzyme 1(IRE-1)αactivator(HG+thapsigargin1μmol/L),and IRE-1αinhibitor(HG+STF-083010,20μmol/L)groups.Additionally,podocytes were divided into 4 groups,including normal,HG,AS-Ⅳ(HG+AS-Ⅳ20μmol/L),and IRE-1αinhibitor(HG+STF-083010,20μmol/L)groups,respectively.After 24 h treatment,the morphology of podocytes and endoplasmic reticulum(ER)was observed by electron microscopy.The expressions of glucose-regulated protein 78(GRP78)and IRE-1αwere detected by cellular immunofluorescence.In in vivo experiment,DN rat model was established via a consecutive 3-day intraperitoneal streptozotocin(STZ)injections.A total of 40 rats were assigned into the normal,DN,AS-Ⅳ[AS-Ⅳ40 mg/(kg·d)],and IRE-1αinhibitor[STF-083010,10 mg/(kg·d)]groups(n=10),respectively.The general condition,24-h urine volume,random blood glucose,urinary protein excretion rate(UAER),blood urea nitrogen(BUN),and serum creatinine(SCr)levels of rats were measured after 8 weeks of intervention.Pathological changes in the renal tissue were observed by hematoxylin and eosin(HE)staining.Quantitative reverse transcription-polymerase chain reaction(RT-PCR)and Western blot were used to detect the expressions of GRP78,IRE-1α,nuclear factor kappa Bp65(NF-κBp65),interleukin(IL)-1β,NLR family pyrin domain containing 3(NLRP3),caspase-1,gasdermin D-N(GSDMD-N),and nephrin at the mRNA and protein levels in vivo and in vitro,respectively.Results:Cytoplasmic vacuolation and ER swelling were observed in the HG and IRE-1αactivator groups.Podocyte morphology and ER expansion were improved in AS-Ⅳand IRE-1αinhibitor groups compared with HG group.Cellular immunofluorescence showed that compared with the normal group,the fluorescence intensity of GRP78 and IRE-1αin the HG and IRE-1αactivator groups were significantly increased whereas decreased in AS-Ⅳand IRE-1αinhibitor groups(P<0.05).Compared with the normal group,the mRNA and protein expressions of GRP78,IRE-1α,NF-κBp65,IL-1β,NLRP3,caspase-1 and GSDMD-N in the HG group was increased(P<0.05).Compared with HG group,the expression of above indices was decreased in the AS-Ⅳand IRE-1αinhibitor groups,and the expression in the IRE-1αactivator group was increased(P<0.05).The expression of nephrin was decreased in the HG group,and increased in AS-Ⅳand IRE-1αinhibitor groups(P<0.05).The in vivo experiment results revealed that compared to the normal group,the levelse of blood glucose,triglyceride,total cholesterol,BUN,SCr and urinary protein in the DN group were higher(P<0.05).Compared with DN group,the above indices in AS-Ⅳand IRE-1αinhibitor groups were decreased(P<0.05).HE staining revealed glomerular hypertrophy,mesangial widening and mesangial cell proliferation in the renal tissue of the DN group.Compared with the DN group,the above pathological changes in renal tissue of AS-Ⅳand IRE-1αinhibitor groups were alleviated.Quantitative RT-PCR and Western blot results of GRP78,IRE-1α,NF-κBp65,IL-1β,NLRP3,caspase-1 and GSDMD-N were consistent with immunofluorescence analysis.Conclusion:AS-Ⅳcould reduce ERS and inflammation,improve podocyte pyroptosis,thus exerting a podocyteprotective effect in DN,through regulating IRE-1α/NF-κB/NLRP3 signaling pathway.
