Background:The 2019 novel coronavirus has caused the outbreak of the acute respiratory disease in Wuhan,Hubei Province of China since December 2019.This study was performed to analyze the clinical characteristics of p...Background:The 2019 novel coronavirus has caused the outbreak of the acute respiratory disease in Wuhan,Hubei Province of China since December 2019.This study was performed to analyze the clinical characteristics of patients who succumbed to and who recovered from 2019 novel coronavirus disease(COVID-19).Methods:Clinical data were collected from two tertiary hospitals in Wuhan.A retrospective investigation was conducted to analyze the clinical characteristics of fatal cases of COVID-19(death group)and we compare them with recovered patients(recovered group).Continuous variables were analyzed using the Mann-WhitneyU test.Categorical variables were analyzed byχ2 test or Fisher exact test as appropriate.Results:Our study enrolled 109 COVID-19 patients who died during hospitalization and 116 recovered patients.The median age of the death group was older than the recovered group(69[62,74]vs.40[33,57]years,Z=9.738,P<0.001).More patients in the death group had underlying diseases(72.5%vs.41.4%,χ2=22.105,P<0.001).Patients in the death group had a significantly longer time of illness onset to hospitalization(10.0[6.5,12.0]vs.7.0[5.0,10.0]days,Z=3.216,P=0.001).On admission,the proportions of patients with symptoms of dyspnea(70.6%vs.19.0%,χ2=60.905,P<0.001)and expectoration(32.1%vs.12.1%,χ2=13.250,P<0.001)were significantly higher in the death group.The blood oxygen saturation was significantly lower in the death group(85[77,91]%vs.97[95,98]%,Z=10.625,P<0.001).The white blood cell(WBC)in death group was significantly higher on admission(7.23[4.87,11.17]vs.4.52[3.62,5.88]×109/L,Z=7.618,P<0.001).Patients in the death group exhibited significantly lower lymphocyte count(0.63[0.40,0.79]vs.1.00[0.72,1.27]×109/L,Z=8.037,P<0.001)and lymphocyte percentage(7.10[4.45,12.73]%vs.23.50[15.27,31.25]%,Z=10.315,P<0.001)on admission,and the lymphocyte percentage continued to decrease during hospitalization(7.10[4.45,12.73]%vs.2.91[1.79,6.13]%,Z=5.242,P<0.001).Alanine transaminase(22.00[15.00,34.00]vs.18.70[13.00,30.38]U/L,Z=2.592,P=0展开更多
【目的】建立在中低温度下直接酶解米糠制备短肽的优化工艺。【方法】采用二次回归正交旋转组合设计优化直接酶解米糠制备短肽的工艺条件,其中总糖含量测定采用蒽酮比色法,水解度(degree of hydrolysis,DH)采用pH-stat法,蛋白质回收率...【目的】建立在中低温度下直接酶解米糠制备短肽的优化工艺。【方法】采用二次回归正交旋转组合设计优化直接酶解米糠制备短肽的工艺条件,其中总糖含量测定采用蒽酮比色法,水解度(degree of hydrolysis,DH)采用pH-stat法,蛋白质回收率采用凯氏定氮法。【结果】确定直接酶解米糠制备短肽最佳工艺条件为:米糠先经糖酶(viscozyme)反应2h去除糖类杂质,然后用碱性蛋白酶(alcalase)和胰蛋白酶双酶水解,最适pH8.2,温度45℃,alcalase与胰蛋白酶酶活比59﹕41,总酶活5750U/g底物,水解时间3h。在此条件下,DH为23.04%,蛋白质回收率为84.33%,酸溶性多肽(TCA-SN)为68.40%,短肽分子量主要集中在1000D以下。【结论】在中低温和偏中性(pH8.2)条件下,采用碱性蛋白酶和胰蛋白酶双酶直接酶解米糠制备短肽,与先提取蛋白后酶解制备短肽的方法相比,具有操作步骤简单、蛋白质利用率高等特点,是一种制备米糠肽的新途径。展开更多
The mechanisms of age-associated memory impairment may be associated with glutamate receptor function and chromatin modification.To observe the effect of an enriched environment on the cognitive function of mice with ...The mechanisms of age-associated memory impairment may be associated with glutamate receptor function and chromatin modification.To observe the effect of an enriched environment on the cognitive function of mice with age-associated memory impairment,3-monthold C57BL/6 male mice("young"mice)were raised in a standard environment,while 24-month-old C57BL/6 male mice with memory impairment("age-associated memory impairment"mice)were raised in either a standard environment or an enriched environment.The enriched environment included a variety of stimuli involving movement and sensation.A water maze test was then used to measure cognitive function in the mice.Furthermore,quantitative real-time polymerase chain reaction and western blot assays were used to detect right hippocampal GluN2B mRNA as well as protein expression of GluN2B and CREB binding protein in all mice.In addition,chromatin immunoprecipitation was used to measure the extent of histone acetylation of the hippocampal GluN2B gene promoters.Compared with the young mice,the water maze performance of age-associated memory impairment mice in the standard environment was significantly decreased.In addition,there were significantly lower levels of total histone acetylation and expression of CREB binding protein in the hippocampus of age-associated memory impairment mice in the standard environment compared with the young mice.There were also significantly lower levels of histone acetylation,protein expression,and mRNA expression of GluN2B in the hippocampus of these mice.In contrast,in the age-associated memory impairment mice with the enriched environment intervention,the water maze performance and molecular biological indexes were significantly improved.These data confirm that an enriched environment can improve cognitive dysfunction in age-associated memory impairment mice,and suggest that the mechanisms may be related to the increased expression of CREB binding protein and the increased degree of total histone acetylation in the hippocampus of age-associated 展开更多
Ischemic stroke can cause blood-brain barrier(BBB)injury,which worsens brain damage induced by stroke.Abnormal expression of tight junction proteins in endothelial cells(ECs)can increase intracellular space and BBB le...Ischemic stroke can cause blood-brain barrier(BBB)injury,which worsens brain damage induced by stroke.Abnormal expression of tight junction proteins in endothelial cells(ECs)can increase intracellular space and BBB leakage.Selective inhibition of mitogen-activated protein kinase,the negative regulatory substrate of mitogen-activated protein kinase phosphatase(MKP)-1,improves tight junction protein function in ECs,and genetic deletion of MKP-1 aggravates ischemic brain injury.However,whether the latter affects BBB integrity,and the cell type-specific mechanism underlying this process,remain unclear.In this study,we established an adult male mouse model of ischemic stroke by occluding the middle cerebral artery for 60 minutes and overexpressed MKP-1 in ECs on the injured side via lentiviral transfection before stroke.We found that overexpression of MKP-1 in ECs reduced infarct volume,reduced the level of inflammatory factors interleukin-1β,interleukin-6,and chemokine C-C motif ligand-2,inhibited vascular injury,and promoted the recovery of sensorimotor and memory/cognitive function.Overexpression of MKP-1 in ECs also inhibited the activation of cerebral ischemia-induced extracellular signal-regulated kinase(ERK)1/2 and the downregulation of occludin expression.Finally,to investigate the mechanism by which MKP-1 exerted these functions in ECs,we established an ischemic stroke model in vitro by depriving the primary endothelial cell of oxygen and glucose,and pharmacologically inhibited the activity of MKP-1 and ERK1/2.Our findings suggest that MKP-1 inhibition aggravates oxygen and glucose deprivation-induced cell death,cell monolayer leakage,and downregulation of occludin expression,and that inhibiting ERK1/2 can reverse these effects.In addition,co-inhibition of MKP-1 and ERK1/2 exhibited similar effects to inhibition of ERK1/2.These findings suggest that overexpression of MKP-1 in ECs can prevent ischemia-induced occludin downregulation and cell death via deactivating ERK1/2,thereby protecting the integrity of BB展开更多
Recent studies in patients with spinal cord injuries(SCIs)have confirmed the diagnostic potential of biofluid-based biomarkers,as a topic of increasing interest in relation to SCI diagnosis and treatment.This paper re...Recent studies in patients with spinal cord injuries(SCIs)have confirmed the diagnostic potential of biofluid-based biomarkers,as a topic of increasing interest in relation to SCI diagnosis and treatment.This paper reviews the research progress and application prospects of recently identified SCI-related biomarkers.Many structural proteins,such as glial fibrillary acidic protein,S100-β,ubiquitin carboxy-terminal hydrolase-L1,neurofilament light,and tau protein were correlated with the diagnosis,American Spinal Injury Association Impairment Scale,and prognosis of SCI to different degrees.Inflammatory factors,including interleukin-6,interleukin-8,and tumor necrosis factorα,are also good biomarkers for the diagnosis of acute and chronic SCI,while non-coding RNAs(micro RNAs and long non-coding RNAs)also show diagnostic potential for SCI.Trace elements(Mg,Se,Cu,Zn)have been shown to be related to motor recovery and can predict motor function after SCI,while humoral markers can reflect the pathophysiological changes after SCI.These factors have the advantages of low cost,convenient sampling,and ease of dynamic tracking,but are also associated with disadvantages,including diverse influencing factors and complex level changes.Although various proteins have been verified as potential biomarkers for SCI,more convincing evidence from large clinical and prospective studies is thus required to identify the most valuable diagnostic and prognostic biomarkers for SCI.展开更多
本文首次利用高黏度壳聚糖(黏度>400 m Pa·s)回收鱼糜漂洗液中的蛋白质,探讨其影响因素,以获得优化的回收工艺条件。通过单因素试验研究了温度、p H、作用时间、壳聚糖浓度对蛋白质回收率的影响,并在此基础上做正交试验,得出优...本文首次利用高黏度壳聚糖(黏度>400 m Pa·s)回收鱼糜漂洗液中的蛋白质,探讨其影响因素,以获得优化的回收工艺条件。通过单因素试验研究了温度、p H、作用时间、壳聚糖浓度对蛋白质回收率的影响,并在此基础上做正交试验,得出优化的试验条件。采用双缩脲法测定蛋白质浓度,重铬酸钾法测定COD。试验结果表明:在优化的试验条件下:pH 6.5,温度20℃,作用时间90 min,壳聚糖质量浓度1.67 mg/mL,高黏度壳聚糖对鱼糜漂洗液中蛋白质回收率达43.68%,COD去除率为22.73%。展开更多
文摘Background:The 2019 novel coronavirus has caused the outbreak of the acute respiratory disease in Wuhan,Hubei Province of China since December 2019.This study was performed to analyze the clinical characteristics of patients who succumbed to and who recovered from 2019 novel coronavirus disease(COVID-19).Methods:Clinical data were collected from two tertiary hospitals in Wuhan.A retrospective investigation was conducted to analyze the clinical characteristics of fatal cases of COVID-19(death group)and we compare them with recovered patients(recovered group).Continuous variables were analyzed using the Mann-WhitneyU test.Categorical variables were analyzed byχ2 test or Fisher exact test as appropriate.Results:Our study enrolled 109 COVID-19 patients who died during hospitalization and 116 recovered patients.The median age of the death group was older than the recovered group(69[62,74]vs.40[33,57]years,Z=9.738,P<0.001).More patients in the death group had underlying diseases(72.5%vs.41.4%,χ2=22.105,P<0.001).Patients in the death group had a significantly longer time of illness onset to hospitalization(10.0[6.5,12.0]vs.7.0[5.0,10.0]days,Z=3.216,P=0.001).On admission,the proportions of patients with symptoms of dyspnea(70.6%vs.19.0%,χ2=60.905,P<0.001)and expectoration(32.1%vs.12.1%,χ2=13.250,P<0.001)were significantly higher in the death group.The blood oxygen saturation was significantly lower in the death group(85[77,91]%vs.97[95,98]%,Z=10.625,P<0.001).The white blood cell(WBC)in death group was significantly higher on admission(7.23[4.87,11.17]vs.4.52[3.62,5.88]×109/L,Z=7.618,P<0.001).Patients in the death group exhibited significantly lower lymphocyte count(0.63[0.40,0.79]vs.1.00[0.72,1.27]×109/L,Z=8.037,P<0.001)and lymphocyte percentage(7.10[4.45,12.73]%vs.23.50[15.27,31.25]%,Z=10.315,P<0.001)on admission,and the lymphocyte percentage continued to decrease during hospitalization(7.10[4.45,12.73]%vs.2.91[1.79,6.13]%,Z=5.242,P<0.001).Alanine transaminase(22.00[15.00,34.00]vs.18.70[13.00,30.38]U/L,Z=2.592,P=0
文摘【目的】建立在中低温度下直接酶解米糠制备短肽的优化工艺。【方法】采用二次回归正交旋转组合设计优化直接酶解米糠制备短肽的工艺条件,其中总糖含量测定采用蒽酮比色法,水解度(degree of hydrolysis,DH)采用pH-stat法,蛋白质回收率采用凯氏定氮法。【结果】确定直接酶解米糠制备短肽最佳工艺条件为:米糠先经糖酶(viscozyme)反应2h去除糖类杂质,然后用碱性蛋白酶(alcalase)和胰蛋白酶双酶水解,最适pH8.2,温度45℃,alcalase与胰蛋白酶酶活比59﹕41,总酶活5750U/g底物,水解时间3h。在此条件下,DH为23.04%,蛋白质回收率为84.33%,酸溶性多肽(TCA-SN)为68.40%,短肽分子量主要集中在1000D以下。【结论】在中低温和偏中性(pH8.2)条件下,采用碱性蛋白酶和胰蛋白酶双酶直接酶解米糠制备短肽,与先提取蛋白后酶解制备短肽的方法相比,具有操作步骤简单、蛋白质利用率高等特点,是一种制备米糠肽的新途径。
基金supported by the National Natural Science Foundation of China,Nos.81672242,81972141(both to YW)the China Postdoctoral Science Fund,No.2017M621675(to XW)+4 种基金the Natural Science Foundation of Jiangsu Province of China,No.BK20171280(to XW)the Six One Project of Scientific Research Project for High-Level Health Talents of Jiangsu Province of China,Nos.LGY2017028,LGY2018027(to XW)the Key Young Medical Talents Project of Jiangsu Province,No.QNRC2016339(to XW)the Yangzhou’s 13th Five-Year Plan for“Ke Jiao Qiang Wei”of China,No.ZDRC65(to XW)the Key Project of Shanghai Science and Technology on Biomedicine of China,No.17411953900(to YW)。
文摘The mechanisms of age-associated memory impairment may be associated with glutamate receptor function and chromatin modification.To observe the effect of an enriched environment on the cognitive function of mice with age-associated memory impairment,3-monthold C57BL/6 male mice("young"mice)were raised in a standard environment,while 24-month-old C57BL/6 male mice with memory impairment("age-associated memory impairment"mice)were raised in either a standard environment or an enriched environment.The enriched environment included a variety of stimuli involving movement and sensation.A water maze test was then used to measure cognitive function in the mice.Furthermore,quantitative real-time polymerase chain reaction and western blot assays were used to detect right hippocampal GluN2B mRNA as well as protein expression of GluN2B and CREB binding protein in all mice.In addition,chromatin immunoprecipitation was used to measure the extent of histone acetylation of the hippocampal GluN2B gene promoters.Compared with the young mice,the water maze performance of age-associated memory impairment mice in the standard environment was significantly decreased.In addition,there were significantly lower levels of total histone acetylation and expression of CREB binding protein in the hippocampus of age-associated memory impairment mice in the standard environment compared with the young mice.There were also significantly lower levels of histone acetylation,protein expression,and mRNA expression of GluN2B in the hippocampus of these mice.In contrast,in the age-associated memory impairment mice with the enriched environment intervention,the water maze performance and molecular biological indexes were significantly improved.These data confirm that an enriched environment can improve cognitive dysfunction in age-associated memory impairment mice,and suggest that the mechanisms may be related to the increased expression of CREB binding protein and the increased degree of total histone acetylation in the hippocampus of age-associated
基金supported by Research Start-up Funding of Shenzhen Traditional Chinese Medicine Hospital,No.2021-07(to FB)Sanming Project of Medicine in Shenzhen,No.SZZYSM 202111011(to XDQ and FB)+1 种基金Key Discipline Established by Zhejiang Province,Jiaxing City Jointly-Pain Medicine,No.2019-ss-ttyx(to LSX)Jiaxing Key Laboratory of Neurology and Pain Medicine,No.[2014]81(to LSX)。
文摘Ischemic stroke can cause blood-brain barrier(BBB)injury,which worsens brain damage induced by stroke.Abnormal expression of tight junction proteins in endothelial cells(ECs)can increase intracellular space and BBB leakage.Selective inhibition of mitogen-activated protein kinase,the negative regulatory substrate of mitogen-activated protein kinase phosphatase(MKP)-1,improves tight junction protein function in ECs,and genetic deletion of MKP-1 aggravates ischemic brain injury.However,whether the latter affects BBB integrity,and the cell type-specific mechanism underlying this process,remain unclear.In this study,we established an adult male mouse model of ischemic stroke by occluding the middle cerebral artery for 60 minutes and overexpressed MKP-1 in ECs on the injured side via lentiviral transfection before stroke.We found that overexpression of MKP-1 in ECs reduced infarct volume,reduced the level of inflammatory factors interleukin-1β,interleukin-6,and chemokine C-C motif ligand-2,inhibited vascular injury,and promoted the recovery of sensorimotor and memory/cognitive function.Overexpression of MKP-1 in ECs also inhibited the activation of cerebral ischemia-induced extracellular signal-regulated kinase(ERK)1/2 and the downregulation of occludin expression.Finally,to investigate the mechanism by which MKP-1 exerted these functions in ECs,we established an ischemic stroke model in vitro by depriving the primary endothelial cell of oxygen and glucose,and pharmacologically inhibited the activity of MKP-1 and ERK1/2.Our findings suggest that MKP-1 inhibition aggravates oxygen and glucose deprivation-induced cell death,cell monolayer leakage,and downregulation of occludin expression,and that inhibiting ERK1/2 can reverse these effects.In addition,co-inhibition of MKP-1 and ERK1/2 exhibited similar effects to inhibition of ERK1/2.These findings suggest that overexpression of MKP-1 in ECs can prevent ischemia-induced occludin downregulation and cell death via deactivating ERK1/2,thereby protecting the integrity of BB
基金financially supported by the National Key Research and Development Project of Stem Cell and Transformational Research,No.2019YFA0112100(to SQF)。
文摘Recent studies in patients with spinal cord injuries(SCIs)have confirmed the diagnostic potential of biofluid-based biomarkers,as a topic of increasing interest in relation to SCI diagnosis and treatment.This paper reviews the research progress and application prospects of recently identified SCI-related biomarkers.Many structural proteins,such as glial fibrillary acidic protein,S100-β,ubiquitin carboxy-terminal hydrolase-L1,neurofilament light,and tau protein were correlated with the diagnosis,American Spinal Injury Association Impairment Scale,and prognosis of SCI to different degrees.Inflammatory factors,including interleukin-6,interleukin-8,and tumor necrosis factorα,are also good biomarkers for the diagnosis of acute and chronic SCI,while non-coding RNAs(micro RNAs and long non-coding RNAs)also show diagnostic potential for SCI.Trace elements(Mg,Se,Cu,Zn)have been shown to be related to motor recovery and can predict motor function after SCI,while humoral markers can reflect the pathophysiological changes after SCI.These factors have the advantages of low cost,convenient sampling,and ease of dynamic tracking,but are also associated with disadvantages,including diverse influencing factors and complex level changes.Although various proteins have been verified as potential biomarkers for SCI,more convincing evidence from large clinical and prospective studies is thus required to identify the most valuable diagnostic and prognostic biomarkers for SCI.
