The non-structural protein 1 is an important molecule of the viruses in flavivirus group including to Zika virus. Recently, the NS1 of Zika virus was discovered. There is still no complete information of the molecular...The non-structural protein 1 is an important molecule of the viruses in flavivirus group including to Zika virus. Recently, the NS1 of Zika virus was discovered. There is still no complete information of the molecular interaction of NS1 of Zika virus which can be the clue for explanation for its pathogenesis and further drug search. Here the authors report the cleft analysis of NS1 of Zika virus and the result can be useful for future development of good diagnostic tool and antiviral drug finding for management of Zika virus.展开更多
HPPD(4-hydroxyphenylpyruvate dioxygenase)inhibitor are widely used in agriculture due to their high efficacy and environmental friendliness.However,many important crops,such as rice,wheat,and soybean,are naturally sen...HPPD(4-hydroxyphenylpyruvate dioxygenase)inhibitor are widely used in agriculture due to their high efficacy and environmental friendliness.However,many important crops,such as rice,wheat,and soybean,are naturally sensitive to these herbicides.In this study,we employed a directed evolution strategy to enhance the metabolic capacity of OsHSL2,OsHSL4,OsHSL6,and SbHSL1 proteins toward HPPD inhibitors,providing a new technological approach as well as theoretical foundation for molecular breeding of herbicide-resistant crops.By combining AlphaFold 3 protein models with crystal structures,we systematically redesigned key residues to resemble the active residues found in HIS1.Catalytic activity assays demonstrated that specific mutations significantly improved the metabolic activity of HSLs proteins toward various HPPD inhibitors.Notably,the OsHSL2-M4 mutant exhibited enhanced metabolic activity for BBC-OH and methyl-benquitrione,while the OsHSL4-M5 mutant completely metabolized BBC-OH and topramezone.Additionally,the SbHSL1-M4 mutant showed significant improvement in the metabolism of BBC-OH and several other herbicides,providing strong evidence to support the use of structure-guided HSL mutations to enhance crop resistance to HPPD inhibitors.展开更多
α-Synuclein accumulation and transmission are vital to the pathogenesis of Parkinson's disease,although the mechanisms underlying misfoldedα-synuclein accumulation and propagation have not been conclusively dete...α-Synuclein accumulation and transmission are vital to the pathogenesis of Parkinson's disease,although the mechanisms underlying misfoldedα-synuclein accumulation and propagation have not been conclusively determined.The expression of low-density lipoprotein receptor–related protein 1,which is abundantly expressed in neurons and considered to be a multifunctional endocytic receptor,is elevated in the neurons of patients with Parkinson's disease.However,whether there is a direct link between low-density lipoprotein receptor–related protein 1 andα-synuclein aggregation and propagation in Parkinson's disease remains unclear.Here,we established animal models of Parkinson's disease by inoculating monkeys and mice withα-synuclein pre-formed fibrils and observed elevated low-density lipoprotein receptor–related protein 1 levels in the striatum and substantia nigra,accompanied by dopaminergic neuron loss and increasedα-synuclein levels.However,low-density lipoprotein receptor–related protein 1 knockdown efficiently rescued dopaminergic neurodegeneration and inhibited the increase inα-synuclein levels in the nigrostriatal system.In HEK293A cells overexpressingα-synuclein fragments,low-density lipoprotein receptor–related protein 1 levels were upregulated only when the N-terminus ofα-synuclein was present,whereas anα-synuclein fragment lacking the N-terminus did not lead to low-density lipoprotein receptor–related protein 1 upregulation.Furthermore,the N-terminus ofα-synuclein was found to be rich in lysine residues,and blocking lysine residues in PC12 cells treated withα-synuclein pre-formed fibrils effectively reduced the elevated low-density lipoprotein receptor–related protein 1 andα-synuclein levels.These findings indicate that low-density lipoprotein receptor–related protein 1 regulates pathological transmission ofα-synuclein from the striatum to the substantia nigra in the nigrostriatal system via lysine residues in theα-synuclein N-terminus.展开更多
Objective: To study protein-protein interaction between heterogeneous nuclear ribonucleoprotein H(hn RNP H) and Dengue virus(DENV) proteins. Methods: DENV proteins were screened against the host hn RNP H protein, in o...Objective: To study protein-protein interaction between heterogeneous nuclear ribonucleoprotein H(hn RNP H) and Dengue virus(DENV) proteins. Methods: DENV proteins were screened against the host hn RNP H protein, in order to identify the host-viral protein-protein interactions in DENV infected THP-1 cells by co-immunoprecipitation. The co-localization of the interacting proteins was further confirmed by immunofluorescence microscopy. Results: The host protein hn RNP H was found to interact with DENV nonstructural 1 protein and help the virus to multiply in the cell. Conclusions: The non-structural 1 glycoprotein is a key modulator of host immune response and is also involved in viral replication. Therefore, disruption of this key interaction between hn RNP H and DENV nonstructural 1 could be an important therapeutic strategy for management of DENV infection.展开更多
Hepatitis C virus (HCV) infection and associated liver diseases are still challenging and represent a significant health care burden around the world. Although, the treatment strategies have been improved by the devel...Hepatitis C virus (HCV) infection and associated liver diseases are still challenging and represent a significant health care burden around the world. Although, the treatment strategies have been improved by the development of novel direct-acting antivirals, but such therapeutic options are still expensive and beyond the financial range of the most infected individuals in developing or even in resource replete countries. It demands an urgent need to search novel and improved alternate treatment strategies to treat the infection. The present study was aimed to develop an in vitro stable cell culture system, persistently expressing HCV genotype 1a non-structural genes and to characterize the inhibitory effects of synthetic siRNAs (short interference RNA) directed against the most conserved regions of nonstructural genes in an in vitro cell culture model. The continuous expression of nonstructural genes for more than 30 days post transfection was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis in stable human hepatoma cell line (Huh-7). The gene expression studies revealed significantly reduced gene expression of HCV nonstructural genes (i.e., NS2, NS4A and NS5A) both at mRNA and protein levels when treated against genome specific synthetic siRNAs in stable cell lines (51%, 47% and 54% respectively, p < 0.05). Similarly, a vivid decrease in HCV viral titer was exhibited by synthetic siRNAs in an in vitro viral replicate cell culture model (58%, 48% and 50%, respectively, p < 0.05) determined by quantitative Real-Time PCR (qPCR). Our data indicate that siRNA mediated gene silencing may be considered a promising alternate treatment strategy against HCV in combination with other effective therapeutic regimens in future.展开更多
目的探究纤维蛋白原样蛋白1(fibrinogen-like protein 1,FGL1)和膜联蛋白A11(annexin A11,ANXA11)对肺癌手术患者效用评估价值及与预后的相关性。方法将本院2020年1月至2023年12月接诊的98例肺癌患者纳入肺癌组,另选取98例健康人作为对...目的探究纤维蛋白原样蛋白1(fibrinogen-like protein 1,FGL1)和膜联蛋白A11(annexin A11,ANXA11)对肺癌手术患者效用评估价值及与预后的相关性。方法将本院2020年1月至2023年12月接诊的98例肺癌患者纳入肺癌组,另选取98例健康人作为对照组。受试者入院后分别于空腹状态下采取6 mL肘静脉血,采用ELISA法检测血清ANXA11水平,采用实时荧光定量PCR技术检测ANXA11表达水平。根据术后1个月肿瘤标志物变化、影像学表现和免疫功能指标变化情况分为治疗有效组(n=68)和无效组(n=30)。患者出院后均行12~36个月的随访,根据患者预后情况分为预后良好组(n=63)和预后不良组(n=35)。对比对照组和肺癌组/有效组和无效组/预后FGL1、ANXA11表达差异;ROC分析FGL1、ANXA11单一及联合检测对肺癌手术患者临床效用评估价值;Spearman分析FGL1、ANXA11表达与预后的关系。结果肺癌组FGL1和ANXA11表达水平明显高于对照组(均P<0.05);无效组FGL1和ANXA11表达水平明显高于有效组(均P<0.05);ROC结果显示FGL1、ANXA11单独及联合检测对肺癌手术患者效用评估的曲线线下面积分别为0.928,并且联合检测具有较高的特异性(95.59%)以及敏感度(90.00%),诊断价值显著高于单独检测ROC曲线线下面积(均P<0.05);预后不良组FGL1和ANXA11表达水平明显高于预后良好组(均P<0.05);Spearman相关性结果显示FGL1、ANXA11与肺癌手术患者预后呈显著正相关(r=0.771、0.793,均P<0.05)。结论肺癌患者的FGL1与ANXA11表达水平高于健康人,治疗无效者高于有效者,预后不良者高于良好者,二者单独及联合检测对患者手术治疗效用评估均有价值,且其与预后相关性显著。展开更多
目的探讨慢性乙型肝炎病毒(hepatitis B virus,HBV)感染相关肝病患者血清壳多糖酶3样蛋白1(chitosan 3-like protein 1,CHI3L1)水平与γ-谷氨酰转肽酶/血小板比值(GPR)和天冬氨酸氨基转移酶/血小板比值(APRI)的相关性及诊断价值。方法...目的探讨慢性乙型肝炎病毒(hepatitis B virus,HBV)感染相关肝病患者血清壳多糖酶3样蛋白1(chitosan 3-like protein 1,CHI3L1)水平与γ-谷氨酰转肽酶/血小板比值(GPR)和天冬氨酸氨基转移酶/血小板比值(APRI)的相关性及诊断价值。方法将纳入的慢性HBV感染者221例,分为慢性乙型肝炎(CHB,n=49)、肝硬化(LC,n=132)和原发性肝癌(PHC,n=40)3组。分别检测血清CHI3L1、丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、γ-谷氨酰转肽酶(γ-GGT)和血小板(PLT),计算GPR及APRI。结果①PHC组年龄、CHI3L1、γ-GGT、GPR及APRI显著高于CHB组(P<0.05),ALT、PLT显著降低(P<0.05);PHC组AST、γ-GGT、GPR及APRI显著高于LC组(P<0.05)。②CHI3L1诊断CHB、LC和PHC的AUC分别为0.748(95%CI:0.640~0.856)、0.933(95%CI:0.896~0.971)和0.928(95%CI:0.860~0.997);GPR诊断CHB、LC和PHC的AUC分别为0.852(95%CI:0.767~0.938)、0.945(95%CI:0.912~0.978)和0.992(95%CI:0.980~1.000);APRI诊断CH组、LC和PHC的AUC分别为0.665(9%CI:0.543~0.786)、0.737(95%CI:0.661~0.813)和0.893(95%CI:0.811~0.974)。③Spearman相关性分析表明,3组患者CHI3L1与GPR、APRI均呈正相关(P<0.05)。结论CHB组、LC组和PHC组中的CHI3L1与GPR的相关性明显优于CHI3L1与APRI的相关性,可用GPR来替代CHI3L1预测肝脏的病变程度。展开更多
文摘The non-structural protein 1 is an important molecule of the viruses in flavivirus group including to Zika virus. Recently, the NS1 of Zika virus was discovered. There is still no complete information of the molecular interaction of NS1 of Zika virus which can be the clue for explanation for its pathogenesis and further drug search. Here the authors report the cleft analysis of NS1 of Zika virus and the result can be useful for future development of good diagnostic tool and antiviral drug finding for management of Zika virus.
基金supported by the National Key Research and Development Program of China(No.2024YFE0214300)Hubei Provincial Science and Technology Plan Project(2022BEC051)selfdetermined research funds of CCNU from the colleges'basic research and operation of MOE(No.CCNU24JCPT023).
文摘HPPD(4-hydroxyphenylpyruvate dioxygenase)inhibitor are widely used in agriculture due to their high efficacy and environmental friendliness.However,many important crops,such as rice,wheat,and soybean,are naturally sensitive to these herbicides.In this study,we employed a directed evolution strategy to enhance the metabolic capacity of OsHSL2,OsHSL4,OsHSL6,and SbHSL1 proteins toward HPPD inhibitors,providing a new technological approach as well as theoretical foundation for molecular breeding of herbicide-resistant crops.By combining AlphaFold 3 protein models with crystal structures,we systematically redesigned key residues to resemble the active residues found in HIS1.Catalytic activity assays demonstrated that specific mutations significantly improved the metabolic activity of HSLs proteins toward various HPPD inhibitors.Notably,the OsHSL2-M4 mutant exhibited enhanced metabolic activity for BBC-OH and methyl-benquitrione,while the OsHSL4-M5 mutant completely metabolized BBC-OH and topramezone.Additionally,the SbHSL1-M4 mutant showed significant improvement in the metabolism of BBC-OH and several other herbicides,providing strong evidence to support the use of structure-guided HSL mutations to enhance crop resistance to HPPD inhibitors.
基金supported by the Natural Science Foundation of Guangxi Zhuang Automomous Region,Nos.2019GXNSFDA245015(to MC),2022GXNSFBA035654(to HL)the National Natural Science Foundation of China,Nos.82360241(to MC),82304876(to HL)+1 种基金Scientific Research and Technology Development Project of Guilin City,Nos.20220139-3(to MC),20210218-5(to HL)Guangxi Medical and Health Key Discipline Construction Project(to QL)。
文摘α-Synuclein accumulation and transmission are vital to the pathogenesis of Parkinson's disease,although the mechanisms underlying misfoldedα-synuclein accumulation and propagation have not been conclusively determined.The expression of low-density lipoprotein receptor–related protein 1,which is abundantly expressed in neurons and considered to be a multifunctional endocytic receptor,is elevated in the neurons of patients with Parkinson's disease.However,whether there is a direct link between low-density lipoprotein receptor–related protein 1 andα-synuclein aggregation and propagation in Parkinson's disease remains unclear.Here,we established animal models of Parkinson's disease by inoculating monkeys and mice withα-synuclein pre-formed fibrils and observed elevated low-density lipoprotein receptor–related protein 1 levels in the striatum and substantia nigra,accompanied by dopaminergic neuron loss and increasedα-synuclein levels.However,low-density lipoprotein receptor–related protein 1 knockdown efficiently rescued dopaminergic neurodegeneration and inhibited the increase inα-synuclein levels in the nigrostriatal system.In HEK293A cells overexpressingα-synuclein fragments,low-density lipoprotein receptor–related protein 1 levels were upregulated only when the N-terminus ofα-synuclein was present,whereas anα-synuclein fragment lacking the N-terminus did not lead to low-density lipoprotein receptor–related protein 1 upregulation.Furthermore,the N-terminus ofα-synuclein was found to be rich in lysine residues,and blocking lysine residues in PC12 cells treated withα-synuclein pre-formed fibrils effectively reduced the elevated low-density lipoprotein receptor–related protein 1 andα-synuclein levels.These findings indicate that low-density lipoprotein receptor–related protein 1 regulates pathological transmission ofα-synuclein from the striatum to the substantia nigra in the nigrostriatal system via lysine residues in theα-synuclein N-terminus.
基金supported by the Defence Institute of Physiology and Allied SciencesDefence Research and Development Organization+1 种基金Ministry of DefenceIndia in the form of TASK-177
文摘Objective: To study protein-protein interaction between heterogeneous nuclear ribonucleoprotein H(hn RNP H) and Dengue virus(DENV) proteins. Methods: DENV proteins were screened against the host hn RNP H protein, in order to identify the host-viral protein-protein interactions in DENV infected THP-1 cells by co-immunoprecipitation. The co-localization of the interacting proteins was further confirmed by immunofluorescence microscopy. Results: The host protein hn RNP H was found to interact with DENV nonstructural 1 protein and help the virus to multiply in the cell. Conclusions: The non-structural 1 glycoprotein is a key modulator of host immune response and is also involved in viral replication. Therefore, disruption of this key interaction between hn RNP H and DENV nonstructural 1 could be an important therapeutic strategy for management of DENV infection.
文摘Hepatitis C virus (HCV) infection and associated liver diseases are still challenging and represent a significant health care burden around the world. Although, the treatment strategies have been improved by the development of novel direct-acting antivirals, but such therapeutic options are still expensive and beyond the financial range of the most infected individuals in developing or even in resource replete countries. It demands an urgent need to search novel and improved alternate treatment strategies to treat the infection. The present study was aimed to develop an in vitro stable cell culture system, persistently expressing HCV genotype 1a non-structural genes and to characterize the inhibitory effects of synthetic siRNAs (short interference RNA) directed against the most conserved regions of nonstructural genes in an in vitro cell culture model. The continuous expression of nonstructural genes for more than 30 days post transfection was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis in stable human hepatoma cell line (Huh-7). The gene expression studies revealed significantly reduced gene expression of HCV nonstructural genes (i.e., NS2, NS4A and NS5A) both at mRNA and protein levels when treated against genome specific synthetic siRNAs in stable cell lines (51%, 47% and 54% respectively, p < 0.05). Similarly, a vivid decrease in HCV viral titer was exhibited by synthetic siRNAs in an in vitro viral replicate cell culture model (58%, 48% and 50%, respectively, p < 0.05) determined by quantitative Real-Time PCR (qPCR). Our data indicate that siRNA mediated gene silencing may be considered a promising alternate treatment strategy against HCV in combination with other effective therapeutic regimens in future.
文摘目的探究纤维蛋白原样蛋白1(fibrinogen-like protein 1,FGL1)和膜联蛋白A11(annexin A11,ANXA11)对肺癌手术患者效用评估价值及与预后的相关性。方法将本院2020年1月至2023年12月接诊的98例肺癌患者纳入肺癌组,另选取98例健康人作为对照组。受试者入院后分别于空腹状态下采取6 mL肘静脉血,采用ELISA法检测血清ANXA11水平,采用实时荧光定量PCR技术检测ANXA11表达水平。根据术后1个月肿瘤标志物变化、影像学表现和免疫功能指标变化情况分为治疗有效组(n=68)和无效组(n=30)。患者出院后均行12~36个月的随访,根据患者预后情况分为预后良好组(n=63)和预后不良组(n=35)。对比对照组和肺癌组/有效组和无效组/预后FGL1、ANXA11表达差异;ROC分析FGL1、ANXA11单一及联合检测对肺癌手术患者临床效用评估价值;Spearman分析FGL1、ANXA11表达与预后的关系。结果肺癌组FGL1和ANXA11表达水平明显高于对照组(均P<0.05);无效组FGL1和ANXA11表达水平明显高于有效组(均P<0.05);ROC结果显示FGL1、ANXA11单独及联合检测对肺癌手术患者效用评估的曲线线下面积分别为0.928,并且联合检测具有较高的特异性(95.59%)以及敏感度(90.00%),诊断价值显著高于单独检测ROC曲线线下面积(均P<0.05);预后不良组FGL1和ANXA11表达水平明显高于预后良好组(均P<0.05);Spearman相关性结果显示FGL1、ANXA11与肺癌手术患者预后呈显著正相关(r=0.771、0.793,均P<0.05)。结论肺癌患者的FGL1与ANXA11表达水平高于健康人,治疗无效者高于有效者,预后不良者高于良好者,二者单独及联合检测对患者手术治疗效用评估均有价值,且其与预后相关性显著。
文摘目的探讨慢性乙型肝炎病毒(hepatitis B virus,HBV)感染相关肝病患者血清壳多糖酶3样蛋白1(chitosan 3-like protein 1,CHI3L1)水平与γ-谷氨酰转肽酶/血小板比值(GPR)和天冬氨酸氨基转移酶/血小板比值(APRI)的相关性及诊断价值。方法将纳入的慢性HBV感染者221例,分为慢性乙型肝炎(CHB,n=49)、肝硬化(LC,n=132)和原发性肝癌(PHC,n=40)3组。分别检测血清CHI3L1、丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、γ-谷氨酰转肽酶(γ-GGT)和血小板(PLT),计算GPR及APRI。结果①PHC组年龄、CHI3L1、γ-GGT、GPR及APRI显著高于CHB组(P<0.05),ALT、PLT显著降低(P<0.05);PHC组AST、γ-GGT、GPR及APRI显著高于LC组(P<0.05)。②CHI3L1诊断CHB、LC和PHC的AUC分别为0.748(95%CI:0.640~0.856)、0.933(95%CI:0.896~0.971)和0.928(95%CI:0.860~0.997);GPR诊断CHB、LC和PHC的AUC分别为0.852(95%CI:0.767~0.938)、0.945(95%CI:0.912~0.978)和0.992(95%CI:0.980~1.000);APRI诊断CH组、LC和PHC的AUC分别为0.665(9%CI:0.543~0.786)、0.737(95%CI:0.661~0.813)和0.893(95%CI:0.811~0.974)。③Spearman相关性分析表明,3组患者CHI3L1与GPR、APRI均呈正相关(P<0.05)。结论CHB组、LC组和PHC组中的CHI3L1与GPR的相关性明显优于CHI3L1与APRI的相关性,可用GPR来替代CHI3L1预测肝脏的病变程度。
