The aim of this study was to investigate the effect of different concentrations of sporamin on the differentiation and proliferation of 3T3-LI preadipocytes, providing the theoretical basis for the development of food...The aim of this study was to investigate the effect of different concentrations of sporamin on the differentiation and proliferation of 3T3-LI preadipocytes, providing the theoretical basis for the development of food to treat obesity and diabetes, The isolation and purification of sporamin from sweet potato species 55-2 were performed by ammonium sulphate precipitation in combination with ion-exchange and gel filtration chromatography. With berberine as a positive control, different concentrations ofsporamin （0.000, 0.125, 0.025, 0.250, 0.500, and 1.000 mg·mL^-1 were used to treat 3T3-L1 preadipocytes. Intracellular fat accumulation and the degree of adipogenesis were quantified using Oil Red O staining and colorimetry, Preadipocytes differentiation was measured by 3（4,5-dimethylthiazolyl-2-yl）-2,5-diphenyltetrazolium bromide （MTT） spectrophotometric assay. Two sporamin proteins, which were separated into sporamin A （31 kD） and sporamin B （22 kD）, could be purified by ion-exchange and gel filtration chromatography. After being treated by different concentrations of sporamin, the differentiation of 3T3-L1 preadipocytes was significantly inhibited, compared with the positive control. When the sporamin solution concentration was 0.500 mg mL-1, the accumulation of lipid droplets within the cells was significantly decreased and the optical density （OD） value of the solution from destained Oil Red O reached to 0.35, which was the lowest value （P〈 0.05）. The proliferation of 3T3-L1 preadipocytes was significantly inhibited by treating at higher sporamin concentrations. In addition, the inhibitory effect was more obvious with the prolonged treatment time （P〈 0.05）. The differentiation and proliferation of 3T3-L1 preadipocytes could be inhibited significantly by the addition of higher concentration sporamin. It was, therefore, suggested that the sporamin was potentially effective for weight loss.展开更多
An important factor in the emergence and progre sion of osteosarcoma(OS)is the dysregulated expression of microRNAs(miRNAs).Transcription factor 7-like 1(TCF7LI),a member of the T cell factor/lymphoid enhancer factor(...An important factor in the emergence and progre sion of osteosarcoma(OS)is the dysregulated expression of microRNAs(miRNAs).Transcription factor 7-like 1(TCF7LI),a member of the T cell factor/lymphoid enhancer factor(TCF/LEF)transcription factor family,interacts with the Wnt signaling pathway regulator β-catenin and acts as a DNA-specific binding protein.This study sought to elucidate the impact of the interaction between miR 3293p and TCF7L1 on.the growth and apoptosis of OS and analyze the regulatory expression relationship between miRNA and mRNA in osteosarcoma cells using a variety of approaches.MiR329-3p was significantly downregulated,while TCF7L1 was considerably up-regulated in all examined OS cell lines.Additionally,a clinical comparison study was performed using the TCGA database.Subsequently,the regulatory relationship between miR-329-3p and TCF7L1 on the proliferation and apoptosis of OS cells was verified through in vitro and in vivo experiments.When miR 329-3p was transfected into the OS cell line,the expression of TCF7L1 decreased,the proliferation of OS cells was inhibited,the cytoskeleton disintegrated,and the nucleus condensed to fom apoptotic bodies.The expression of proteins that indicate apoptosis increased simultaneously.The cell cycle was arrested in the G0/G1 phase,and the G1/S transition was blocked.The introduction of miR 3293p also inhibited downstream Cyclin D1 of the Wnt pathway.Xenograf experiments indicated that the overexpression of miR-329-3p signi ficanly inhibited the growth of OS xenografts in nude mice,and the expression of TCF7L1 and C-Myc in tumor tssues decreased.MiR 329-3p was significantly reduced in OS cells and played a suppressive role in tumorigenesis and proliferation by targeting TCF7L1 both in vitro and in vivo.Osteosarcoma cell cycle arrest and pathway inhibition were observed upon the regulation of TCF7LI by miR 3293p.Summarizing these results,it can be inferred that miR.3293p exerts anticancer efects in osteosarcoma by inhibiting TCF7L1.展开更多
目的探讨(1-3)-β-D葡聚糖联合降钙素原(procalcitonin,PCT)、CD4^(+)T淋巴细胞多指标在艾滋病患者马尔尼菲篮状菌感染早期诊断临床研究。方法回顾性选取我院2020年1月—2022年6月住院的120例艾滋病患者为研究对象。依据实验室结果,将...目的探讨(1-3)-β-D葡聚糖联合降钙素原(procalcitonin,PCT)、CD4^(+)T淋巴细胞多指标在艾滋病患者马尔尼菲篮状菌感染早期诊断临床研究。方法回顾性选取我院2020年1月—2022年6月住院的120例艾滋病患者为研究对象。依据实验室结果,将其分为马尔尼菲篮状菌感染确诊组(血或组织液培育养出马尔尼菲篮状菌),简称A组(62例),及马尔尼菲篮状菌感染临床诊断组[根据临床症状、体征、血常规及(1-3)-β-D葡聚糖、PCT、CD4^(+)T淋巴细胞多指标诊断],简称B组(58例)。检测患者(1-3)-β-D葡聚糖、PCT、CD4^(+)T淋巴细胞的表达水平,采用受试者工作特征(receiver-operating characteristic,ROC)曲线下面积(area under the curve,AUC)评估上述指标联合检测对艾滋病患者感染马尔尼菲篮状菌的诊断效能。结果A组的(1-3)-β-D葡聚糖和PCT水平均高于B组,CD4^(+)T淋巴细胞个数低于B组(P<0.05);(1-3)-β-D葡聚糖、PCT、CD4^(+)T淋巴细胞联合检测的AUC为0.933,(1-3)-β-D葡聚糖单独检测的AUC是0.812,PCT单独检测的AUC为0.883,CD4^(+)T淋巴细胞单独检测的AUC是0.810,(1-3)-β-D葡聚糖、PCT和CD4^(+)T淋巴细胞联合检测的AUC皆优于三项单独检测,表明(1-3)-β-D葡聚糖、PCT和CD4^(+)T淋巴细胞联合检测的诊断价值皆优于单一指标诊断,且联合检测的特异度、约登指数分别为92.43%和0.580,均高于三项单独检测。结论(1-3)-β-D葡聚糖联合PCT和CD4^(+)T淋巴细胞多指标对艾滋病马尔尼菲篮状菌感染具有非常高的临床诊断价值,能够帮助医生分析出高危风险患者,及时制定治疗方案,同时也承担预后效果的判断依据,对治疗艾滋病马尔尼菲篮状菌感染具有非常重要的研究价值。展开更多
Objective： To investigate the effect of the calcium channel blocker verapamil on adipocyte differentiation and its mechanism of action. Methods： Preadipocytes from 3T3-L1 strain mouse embryos were cultured and diffe...Objective： To investigate the effect of the calcium channel blocker verapamil on adipocyte differentiation and its mechanism of action. Methods： Preadipocytes from 3T3-L1 strain mouse embryos were cultured and differentiated into matured adipocytes in vitro. Verapamil was added to the culture medium in the concentration of 30 μmol/L on Day 0. Cell differentiation was determined by Oil Red O staining and marker gene mRNA expression was evaluated and compared by RT-PCR. The fluo-3/AM probe and laser scanning confocal microscopy were used to measure intracetlular calcium concentrations. Results： （1）The differentiation rate of 3T3-L1 preadipocytes exposed to verapamil was lower than that of untreated cells. （2）Verapamil promoted the retention of pref-1 gene expression. Lipoprotein lipase expression in the verapamil group was significantly lower than that in the control group on Day 4, Day 6 and Day 8 （P 〈 0.05） and resistin expression was significantly lower than that in the control group on Day 6, Day 8 and Day 10 （P 〈 0.05）. Fatty acid synthase expression in the verapamil group was significantly lower than that in the control group from Day 2 （P 〈 0.05）. （3） Intracellular concentrations of calcium [Ca^2＋]i in the verapamil group were significantly decreased compared with those in the control group on Day 2, Day 4 and Day 6 （P 〈 0.05）, while there was no obvious difference between the two groups on Day 0 （P 〉 0.05）. Conclusion： In 3T3-L1 preadipocytes verapamil significantly reduced adipocyte differentiation, down-regulated the mRNA expression of three marker genes for adipocytes differentiation, and prolonged the mRNA expression of an inhibitor of differentiation. The inhibitory effect of verapamil on differentiation may involve its role as a blocker of calcium influx in adipocytes.展开更多
目的探讨血清细胞因子信号转导负调控因子-3(suppressor of cytokine signaling 3,SOCS-3)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-16(interleukin-16,IL-16)及T辅助细胞1/T辅助细胞2(Th1/Th2)与妊娠期高血压...目的探讨血清细胞因子信号转导负调控因子-3(suppressor of cytokine signaling 3,SOCS-3)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-16(interleukin-16,IL-16)及T辅助细胞1/T辅助细胞2(Th1/Th2)与妊娠期高血压疾病(HDCP)的关系。方法选取2014年1月-2016年6月本院妇产科收治的妊娠期高血压疾病患者182例,其中妊娠期高血压组51例,轻度子痫前期组69例,重度子痫前期组62例。另选择同期分娩健康孕妇65例作为对照组。采用ELISA法检测各组孕妇血清SOCS-3、TNF-α、IL-16及Th1/Th2水平,分析重度子痫前期患者不良妊娠结局与各指标的关系。采用Spearman相关分析各组HDCP患者血清SOCS-3水平与TNF-α、IL-16及Th1/Th2的相关性。结果与对照组比较,轻度子痫前期组和重度子痫前期组血清SOCS-3水平均明显降低(0.59±0.12 vs 0.14±0.03和0.08±0.01;P均<0.05)。各组TNF-α、IL-16及Th1/Th2水平均高于对照组,且重度子痫前期组较轻度子痫前期组升高更明显[TNF-α(ng/L):12.58±2.37 vs 34.75±6.42和61.53±9.26;IL-16(ng/L):108.47±35.24 vs 187.63±81.47和284.62±113.58;Th1/Th2:9.27±2.38 vs20.15±4.82和26.48±6.13;P均<0.05]。重度子痫前期患者中,SOCS-3低表达组胎儿宫内窘迫综合征、胎儿生长受限及新生儿窒息的发生率均显著高于高表达组(P均<0.05),TNF-α、IL-16、Th1/Th2高表达组早产及胎儿生长受限的发生率均显著高于低表达组(P均<0.05)。相关分析显示,妊娠期高血压患者血清SOCS-3水平与TNF-α、IL-16、Th1/Th2均呈负相关(r=-0.528,r=-0.506,r=-0.126;P均<0.05)。结论监测孕妇血清SOCS-3、TNF-α、IL-16及Th1/Th2对妊娠期高血压疾病尤其是重度子痫前期的早发现、早诊断、早治疗具有一定的临床指导意义。展开更多
基金supported by the National 863 Program of China (2006AA10Z332)the Ear Marked Fund for Modern Agro-Industry Technology Research System, China
文摘The aim of this study was to investigate the effect of different concentrations of sporamin on the differentiation and proliferation of 3T3-LI preadipocytes, providing the theoretical basis for the development of food to treat obesity and diabetes, The isolation and purification of sporamin from sweet potato species 55-2 were performed by ammonium sulphate precipitation in combination with ion-exchange and gel filtration chromatography. With berberine as a positive control, different concentrations ofsporamin （0.000, 0.125, 0.025, 0.250, 0.500, and 1.000 mg·mL^-1 were used to treat 3T3-L1 preadipocytes. Intracellular fat accumulation and the degree of adipogenesis were quantified using Oil Red O staining and colorimetry, Preadipocytes differentiation was measured by 3（4,5-dimethylthiazolyl-2-yl）-2,5-diphenyltetrazolium bromide （MTT） spectrophotometric assay. Two sporamin proteins, which were separated into sporamin A （31 kD） and sporamin B （22 kD）, could be purified by ion-exchange and gel filtration chromatography. After being treated by different concentrations of sporamin, the differentiation of 3T3-L1 preadipocytes was significantly inhibited, compared with the positive control. When the sporamin solution concentration was 0.500 mg mL-1, the accumulation of lipid droplets within the cells was significantly decreased and the optical density （OD） value of the solution from destained Oil Red O reached to 0.35, which was the lowest value （P〈 0.05）. The proliferation of 3T3-L1 preadipocytes was significantly inhibited by treating at higher sporamin concentrations. In addition, the inhibitory effect was more obvious with the prolonged treatment time （P〈 0.05）. The differentiation and proliferation of 3T3-L1 preadipocytes could be inhibited significantly by the addition of higher concentration sporamin. It was, therefore, suggested that the sporamin was potentially effective for weight loss.
基金The Fund of National Cancer Center Research and Development(26-A-4),The Grants-in-Aid for Scientific Research(Grant Nos.15K10451,16K10866 and 16K20063)from Japan Society for the Promotion of Science.
文摘An important factor in the emergence and progre sion of osteosarcoma(OS)is the dysregulated expression of microRNAs(miRNAs).Transcription factor 7-like 1(TCF7LI),a member of the T cell factor/lymphoid enhancer factor(TCF/LEF)transcription factor family,interacts with the Wnt signaling pathway regulator β-catenin and acts as a DNA-specific binding protein.This study sought to elucidate the impact of the interaction between miR 3293p and TCF7L1 on.the growth and apoptosis of OS and analyze the regulatory expression relationship between miRNA and mRNA in osteosarcoma cells using a variety of approaches.MiR329-3p was significantly downregulated,while TCF7L1 was considerably up-regulated in all examined OS cell lines.Additionally,a clinical comparison study was performed using the TCGA database.Subsequently,the regulatory relationship between miR-329-3p and TCF7L1 on the proliferation and apoptosis of OS cells was verified through in vitro and in vivo experiments.When miR 329-3p was transfected into the OS cell line,the expression of TCF7L1 decreased,the proliferation of OS cells was inhibited,the cytoskeleton disintegrated,and the nucleus condensed to fom apoptotic bodies.The expression of proteins that indicate apoptosis increased simultaneously.The cell cycle was arrested in the G0/G1 phase,and the G1/S transition was blocked.The introduction of miR 3293p also inhibited downstream Cyclin D1 of the Wnt pathway.Xenograf experiments indicated that the overexpression of miR-329-3p signi ficanly inhibited the growth of OS xenografts in nude mice,and the expression of TCF7L1 and C-Myc in tumor tssues decreased.MiR 329-3p was significantly reduced in OS cells and played a suppressive role in tumorigenesis and proliferation by targeting TCF7L1 both in vitro and in vivo.Osteosarcoma cell cycle arrest and pathway inhibition were observed upon the regulation of TCF7LI by miR 3293p.Summarizing these results,it can be inferred that miR.3293p exerts anticancer efects in osteosarcoma by inhibiting TCF7L1.
文摘目的探讨(1-3)-β-D葡聚糖联合降钙素原(procalcitonin,PCT)、CD4^(+)T淋巴细胞多指标在艾滋病患者马尔尼菲篮状菌感染早期诊断临床研究。方法回顾性选取我院2020年1月—2022年6月住院的120例艾滋病患者为研究对象。依据实验室结果,将其分为马尔尼菲篮状菌感染确诊组(血或组织液培育养出马尔尼菲篮状菌),简称A组(62例),及马尔尼菲篮状菌感染临床诊断组[根据临床症状、体征、血常规及(1-3)-β-D葡聚糖、PCT、CD4^(+)T淋巴细胞多指标诊断],简称B组(58例)。检测患者(1-3)-β-D葡聚糖、PCT、CD4^(+)T淋巴细胞的表达水平,采用受试者工作特征(receiver-operating characteristic,ROC)曲线下面积(area under the curve,AUC)评估上述指标联合检测对艾滋病患者感染马尔尼菲篮状菌的诊断效能。结果A组的(1-3)-β-D葡聚糖和PCT水平均高于B组,CD4^(+)T淋巴细胞个数低于B组(P<0.05);(1-3)-β-D葡聚糖、PCT、CD4^(+)T淋巴细胞联合检测的AUC为0.933,(1-3)-β-D葡聚糖单独检测的AUC是0.812,PCT单独检测的AUC为0.883,CD4^(+)T淋巴细胞单独检测的AUC是0.810,(1-3)-β-D葡聚糖、PCT和CD4^(+)T淋巴细胞联合检测的AUC皆优于三项单独检测,表明(1-3)-β-D葡聚糖、PCT和CD4^(+)T淋巴细胞联合检测的诊断价值皆优于单一指标诊断,且联合检测的特异度、约登指数分别为92.43%和0.580,均高于三项单独检测。结论(1-3)-β-D葡聚糖联合PCT和CD4^(+)T淋巴细胞多指标对艾滋病马尔尼菲篮状菌感染具有非常高的临床诊断价值,能够帮助医生分析出高危风险患者,及时制定治疗方案,同时也承担预后效果的判断依据,对治疗艾滋病马尔尼菲篮状菌感染具有非常重要的研究价值。
基金supported by grants from the National Natural Science Foundation of China (30371502)the Natural Science Foundation of Jiangsu Province (BK2001120)
文摘Objective： To investigate the effect of the calcium channel blocker verapamil on adipocyte differentiation and its mechanism of action. Methods： Preadipocytes from 3T3-L1 strain mouse embryos were cultured and differentiated into matured adipocytes in vitro. Verapamil was added to the culture medium in the concentration of 30 μmol/L on Day 0. Cell differentiation was determined by Oil Red O staining and marker gene mRNA expression was evaluated and compared by RT-PCR. The fluo-3/AM probe and laser scanning confocal microscopy were used to measure intracetlular calcium concentrations. Results： （1）The differentiation rate of 3T3-L1 preadipocytes exposed to verapamil was lower than that of untreated cells. （2）Verapamil promoted the retention of pref-1 gene expression. Lipoprotein lipase expression in the verapamil group was significantly lower than that in the control group on Day 4, Day 6 and Day 8 （P 〈 0.05） and resistin expression was significantly lower than that in the control group on Day 6, Day 8 and Day 10 （P 〈 0.05）. Fatty acid synthase expression in the verapamil group was significantly lower than that in the control group from Day 2 （P 〈 0.05）. （3） Intracellular concentrations of calcium [Ca^2＋]i in the verapamil group were significantly decreased compared with those in the control group on Day 2, Day 4 and Day 6 （P 〈 0.05）, while there was no obvious difference between the two groups on Day 0 （P 〉 0.05）. Conclusion： In 3T3-L1 preadipocytes verapamil significantly reduced adipocyte differentiation, down-regulated the mRNA expression of three marker genes for adipocytes differentiation, and prolonged the mRNA expression of an inhibitor of differentiation. The inhibitory effect of verapamil on differentiation may involve its role as a blocker of calcium influx in adipocytes.
文摘目的探讨血清细胞因子信号转导负调控因子-3(suppressor of cytokine signaling 3,SOCS-3)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-16(interleukin-16,IL-16)及T辅助细胞1/T辅助细胞2(Th1/Th2)与妊娠期高血压疾病(HDCP)的关系。方法选取2014年1月-2016年6月本院妇产科收治的妊娠期高血压疾病患者182例,其中妊娠期高血压组51例,轻度子痫前期组69例,重度子痫前期组62例。另选择同期分娩健康孕妇65例作为对照组。采用ELISA法检测各组孕妇血清SOCS-3、TNF-α、IL-16及Th1/Th2水平,分析重度子痫前期患者不良妊娠结局与各指标的关系。采用Spearman相关分析各组HDCP患者血清SOCS-3水平与TNF-α、IL-16及Th1/Th2的相关性。结果与对照组比较,轻度子痫前期组和重度子痫前期组血清SOCS-3水平均明显降低(0.59±0.12 vs 0.14±0.03和0.08±0.01;P均<0.05)。各组TNF-α、IL-16及Th1/Th2水平均高于对照组,且重度子痫前期组较轻度子痫前期组升高更明显[TNF-α(ng/L):12.58±2.37 vs 34.75±6.42和61.53±9.26;IL-16(ng/L):108.47±35.24 vs 187.63±81.47和284.62±113.58;Th1/Th2:9.27±2.38 vs20.15±4.82和26.48±6.13;P均<0.05]。重度子痫前期患者中,SOCS-3低表达组胎儿宫内窘迫综合征、胎儿生长受限及新生儿窒息的发生率均显著高于高表达组(P均<0.05),TNF-α、IL-16、Th1/Th2高表达组早产及胎儿生长受限的发生率均显著高于低表达组(P均<0.05)。相关分析显示,妊娠期高血压患者血清SOCS-3水平与TNF-α、IL-16、Th1/Th2均呈负相关(r=-0.528,r=-0.506,r=-0.126;P均<0.05)。结论监测孕妇血清SOCS-3、TNF-α、IL-16及Th1/Th2对妊娠期高血压疾病尤其是重度子痫前期的早发现、早诊断、早治疗具有一定的临床指导意义。
