Islets from RIP-PDE3B mice, exhibiting β-cell specific overexpression of the cAMP/cGMP-degrading enzyme phosphodiesterase 3B (PDE3B) and dysregulated insulin secretion, were subjected to microarray analysis. We show ...Islets from RIP-PDE3B mice, exhibiting β-cell specific overexpression of the cAMP/cGMP-degrading enzyme phosphodiesterase 3B (PDE3B) and dysregulated insulin secretion, were subjected to microarray analysis. We show that osteopontin (OPN) mRNA is increased in a dose-dependent manner in islets from RIP-PDE3B mice, as compared to wild-type islets. In addition, in silico analysis shows that PDE3B and OPN are interacting. Furthermore, OPN interacts with protein kinase CK2 ina distinct submodule of the protein-protein interaction network. We studied PDE3B and OPN proteins and, in some cases, also PDE1B and PDE4C, under conditions of relevance for insulin secretion. In the presence of forskolin, PDE inhibitors, insulin, or a protein kinase CK2 inhibitor, similar alterations in protein levels of PDE3B and OPN are shown. In summary, results from using a number of strategies demonstrate a connection between PDE3B and OPNas well as a role for protein kinase CK2 inpancreatic β-cells.展开更多
Aneurysmal subarachnoid hemorrhage remains devastating,and the most important determinant of poor outcome is early brain injury（EBI）.In clinical settings,as a surrogate marker of EBI,loss of consciousness at ictus,p...Aneurysmal subarachnoid hemorrhage remains devastating,and the most important determinant of poor outcome is early brain injury（EBI）.In clinical settings,as a surrogate marker of EBI,loss of consciousness at ictus,poor initial clinical grades,and some radiographic findings are used,but these markers are somewhat subjective.Thus,it is imperative to find biomarkers of EBI that have beneficial prognostic and therapeutic implications.In our opinion,an ideal biomarker is a molecule that is implicated in the pathogenesis of both EBI and subsequently developing delayed cerebral ischemia（DCI）,being a therapeutic target,and can be measured easily in the peripheral blood in an acute stage.A good candidate of such a biomarker is a matricellular protein,which is a secreted,inducible and multifunctional extracellular matrix protein.There are many kinds of matricellular proteins reported,but only tenascin-C,osteopontin,galectin-3 and periostin are reported relevant to EBI and DCI.Reliable biomarkers of EBI may stratify aneurysmal subarachnoid hemorrhage patients into categories of risk to develop DCI,and allow objective monitoring of the response to treatment for EBI and earlier diagnosis of DCI.This review emphasizes that further investigation of matricellular proteins as an avenue for biomarker discovery is warranted.展开更多
Objective:To analyze mRNA and protein expression of osteopontin(OPN)in human endome-trium throughout menstrual cycle.Methods:Immunohistochemieal method was used to determine the level and the location ofOPN in normal ...Objective:To analyze mRNA and protein expression of osteopontin(OPN)in human endome-trium throughout menstrual cycle.Methods:Immunohistochemieal method was used to determine the level and the location ofOPN in normal cycling endometrium of 50 women.Western Blot analysis was also used to detectthe existence of OPN at proliferative and secretory phases in six samples.The levels of OPNmRNA in 18 samples were measured by RT-PCR.Results:The OPN mainly expressed in human endometrial glandular epithelium but not instromal cells.The expression was highest at mid and late secretory phases and menstruous phaseas well.No expression was found in early and mid proliferative endometria.Two subtypes ofOPN proteins(40 kDa-75 kDa)were detected by Western blot analysis in homogenized endome-tria at secretory phase.The level of OPN mRNA in secretory phase endometrium was significantlyhigher than that in proliferative phase.Conclusions:OPN was only found in endometrial glandular epithelial cells,and the OPN andits mRNA showed a pronounced cycle-dependent expression in human endometrium,higher ex-pression at mid and late secretory phases and menstruous phase.It's probably involved in embry-onic implantation and endometrial shedding.展开更多
目的:研究在纳米细菌诱导实验性大鼠肾结石形成过程中,大鼠肠道草酸降解菌的改变及其与肾结石形成的关系。方法:30只4周龄Wistar雄性大鼠适应性喂养1周后,按照随机数表法分为三组,对照组(NC组):1.2 ml 0.9%氯化钠溶液尾静脉注射+2 ml 0...目的:研究在纳米细菌诱导实验性大鼠肾结石形成过程中,大鼠肠道草酸降解菌的改变及其与肾结石形成的关系。方法:30只4周龄Wistar雄性大鼠适应性喂养1周后,按照随机数表法分为三组,对照组(NC组):1.2 ml 0.9%氯化钠溶液尾静脉注射+2 ml 0.9%氯化钠溶液灌胃;干扰组(NBT组):1.2 ml纳米细菌悬液尾静脉注射+2 ml四环素溶液灌胃;纳米细菌组(NB组):1.2 ml纳米细菌悬液尾静脉注射+2 ml 0.9%氯化钠溶液灌胃。标准饲养8周后处死,取双侧肾脏组织行HE、Pizzolatto’s染色,称取1 g回盲部新鲜粪便并提取DNA,使用qPCR法检测产甲酸草酸杆菌(Oxc)、乳杆菌属细菌(Lac)、双歧杆菌属细菌(Bif)和肠杆菌科细菌(Ent)四种主要肠道草酸降解细菌的基因表达量,留取腹主动脉血液及肾脏组织,流式细胞仪检测淋巴细胞分化情况,WB检测骨桥蛋白(OPN)及Tamm-Horsfall蛋白(THP)两种炎性蛋白在肾脏组织中的表达情况。结果:NC组肾小管管腔内未见明显晶体及炎症细胞浸润,NBT组部分肾小管扩张,肾小管管腔内可见不规则晶体及炎症细胞浸润,NB组肾小管明显扩张,肾小管管腔内可见大量不规则晶体,部分连接成片,可见大量炎症细胞浸润;三组大鼠粪便中均检测到四种肠道草酸降解菌,且各肠道草酸降解菌在三组大鼠盲肠中的占比相似,均为Oxc最高,约占半数,Lac、Ent次之,Bif最少,仅占约1%,三组大鼠粪便中各肠道草酸降解菌占比无统计学差异(均P>0.05),但三组粪便中肠道草酸降解菌的总量有统计学差异,NC组最高、NB组次之、NBT组最低,NC组与其余两组间的差异均有统计学意义(均P<0.01);流式细胞筛选结果提示三组大鼠血液淋巴细胞出现不同程度分化,Pearson相关性分析结果提示OPN与THP联系紧密,增长趋势具有一致性并且THP蛋白与CD4^(+)、CD8^(+)T淋巴细胞也具有强相关性。结论:纳米细菌诱导肾结石形成过程中肠道草酸降解菌总量降低,但各肠道草酸降解菌所占比例无明显改变。展开更多
文摘Islets from RIP-PDE3B mice, exhibiting β-cell specific overexpression of the cAMP/cGMP-degrading enzyme phosphodiesterase 3B (PDE3B) and dysregulated insulin secretion, were subjected to microarray analysis. We show that osteopontin (OPN) mRNA is increased in a dose-dependent manner in islets from RIP-PDE3B mice, as compared to wild-type islets. In addition, in silico analysis shows that PDE3B and OPN are interacting. Furthermore, OPN interacts with protein kinase CK2 ina distinct submodule of the protein-protein interaction network. We studied PDE3B and OPN proteins and, in some cases, also PDE1B and PDE4C, under conditions of relevance for insulin secretion. In the presence of forskolin, PDE inhibitors, insulin, or a protein kinase CK2 inhibitor, similar alterations in protein levels of PDE3B and OPN are shown. In summary, results from using a number of strategies demonstrate a connection between PDE3B and OPNas well as a role for protein kinase CK2 inpancreatic β-cells.
基金supported by a Grant-in-Aid for Scientific Research from Novartis Pharmaceuticals to HS
文摘Aneurysmal subarachnoid hemorrhage remains devastating,and the most important determinant of poor outcome is early brain injury（EBI）.In clinical settings,as a surrogate marker of EBI,loss of consciousness at ictus,poor initial clinical grades,and some radiographic findings are used,but these markers are somewhat subjective.Thus,it is imperative to find biomarkers of EBI that have beneficial prognostic and therapeutic implications.In our opinion,an ideal biomarker is a molecule that is implicated in the pathogenesis of both EBI and subsequently developing delayed cerebral ischemia（DCI）,being a therapeutic target,and can be measured easily in the peripheral blood in an acute stage.A good candidate of such a biomarker is a matricellular protein,which is a secreted,inducible and multifunctional extracellular matrix protein.There are many kinds of matricellular proteins reported,but only tenascin-C,osteopontin,galectin-3 and periostin are reported relevant to EBI and DCI.Reliable biomarkers of EBI may stratify aneurysmal subarachnoid hemorrhage patients into categories of risk to develop DCI,and allow objective monitoring of the response to treatment for EBI and earlier diagnosis of DCI.This review emphasizes that further investigation of matricellular proteins as an avenue for biomarker discovery is warranted.
文摘Objective:To analyze mRNA and protein expression of osteopontin(OPN)in human endome-trium throughout menstrual cycle.Methods:Immunohistochemieal method was used to determine the level and the location ofOPN in normal cycling endometrium of 50 women.Western Blot analysis was also used to detectthe existence of OPN at proliferative and secretory phases in six samples.The levels of OPNmRNA in 18 samples were measured by RT-PCR.Results:The OPN mainly expressed in human endometrial glandular epithelium but not instromal cells.The expression was highest at mid and late secretory phases and menstruous phaseas well.No expression was found in early and mid proliferative endometria.Two subtypes ofOPN proteins(40 kDa-75 kDa)were detected by Western blot analysis in homogenized endome-tria at secretory phase.The level of OPN mRNA in secretory phase endometrium was significantlyhigher than that in proliferative phase.Conclusions:OPN was only found in endometrial glandular epithelial cells,and the OPN andits mRNA showed a pronounced cycle-dependent expression in human endometrium,higher ex-pression at mid and late secretory phases and menstruous phase.It's probably involved in embry-onic implantation and endometrial shedding.
文摘目的:研究在纳米细菌诱导实验性大鼠肾结石形成过程中,大鼠肠道草酸降解菌的改变及其与肾结石形成的关系。方法:30只4周龄Wistar雄性大鼠适应性喂养1周后,按照随机数表法分为三组,对照组(NC组):1.2 ml 0.9%氯化钠溶液尾静脉注射+2 ml 0.9%氯化钠溶液灌胃;干扰组(NBT组):1.2 ml纳米细菌悬液尾静脉注射+2 ml四环素溶液灌胃;纳米细菌组(NB组):1.2 ml纳米细菌悬液尾静脉注射+2 ml 0.9%氯化钠溶液灌胃。标准饲养8周后处死,取双侧肾脏组织行HE、Pizzolatto’s染色,称取1 g回盲部新鲜粪便并提取DNA,使用qPCR法检测产甲酸草酸杆菌(Oxc)、乳杆菌属细菌(Lac)、双歧杆菌属细菌(Bif)和肠杆菌科细菌(Ent)四种主要肠道草酸降解细菌的基因表达量,留取腹主动脉血液及肾脏组织,流式细胞仪检测淋巴细胞分化情况,WB检测骨桥蛋白(OPN)及Tamm-Horsfall蛋白(THP)两种炎性蛋白在肾脏组织中的表达情况。结果:NC组肾小管管腔内未见明显晶体及炎症细胞浸润,NBT组部分肾小管扩张,肾小管管腔内可见不规则晶体及炎症细胞浸润,NB组肾小管明显扩张,肾小管管腔内可见大量不规则晶体,部分连接成片,可见大量炎症细胞浸润;三组大鼠粪便中均检测到四种肠道草酸降解菌,且各肠道草酸降解菌在三组大鼠盲肠中的占比相似,均为Oxc最高,约占半数,Lac、Ent次之,Bif最少,仅占约1%,三组大鼠粪便中各肠道草酸降解菌占比无统计学差异(均P>0.05),但三组粪便中肠道草酸降解菌的总量有统计学差异,NC组最高、NB组次之、NBT组最低,NC组与其余两组间的差异均有统计学意义(均P<0.01);流式细胞筛选结果提示三组大鼠血液淋巴细胞出现不同程度分化,Pearson相关性分析结果提示OPN与THP联系紧密,增长趋势具有一致性并且THP蛋白与CD4^(+)、CD8^(+)T淋巴细胞也具有强相关性。结论:纳米细菌诱导肾结石形成过程中肠道草酸降解菌总量降低,但各肠道草酸降解菌所占比例无明显改变。
