Clip domain serine proteases （cSPs） and their homologs （SPHs） play an important role in various biological processes that are essential components of extracellular signaling cascades, especially in the innate immu...Clip domain serine proteases （cSPs） and their homologs （SPHs） play an important role in various biological processes that are essential components of extracellular signaling cascades, especially in the innate immune responses of invertebrates. Here, polymorphisms of PtcSP and PtSPH from the swimming crab Portunus tritubereulatus were investigated to explore their association with resistance/ susceptibility to Vibrio alginolyticus. Polymorphic loci were identified using Clustal X, and characterized with SPSS 16.0 software, and then the significance of genotype and allele frequencies between resistant and susceptible stocks was determined by a Zz test. A total of 109 and 77 single nucleotide polymorphisms （SNPs） were identified in the genomic fragments of PtcSP and PtSPH, respectively. Notably, nearly half of PtSPH polymorphisms were found in the non-coding exon 1. Fourteen SNPs investigated were significantly associated with susceptibility/resistance to I1. alginolyticus （P〈0.05）. Among them, eight SNPs were observed in introns, and one synonymous, four non-synonymous SNPs and one ins-del were found in coding exons. In addition, five simple sequence repeats （SSRs） were detected in intron 3 of PtcSP. Although there was no statistically significant difference of allele frequencies, the SSRs showed different polymorphic alleles on the basis of the repeat number between resistant and susceptible stocks. After fiarther validation, polymorphisms investigated here might be applied to select potential molecular markers ofP. trituberculatus with resistance to I1. alginolyticus.展开更多
Emiliania huxleyi is the most prominent modern coccolithophore,a group of marine unicellular eukaryotes that play a critical role in ocean biogeochemistry.Coccolithoviruses are large double stranded DNA viruses,which ...Emiliania huxleyi is the most prominent modern coccolithophore,a group of marine unicellular eukaryotes that play a critical role in ocean biogeochemistry.Coccolithoviruses are large double stranded DNA viruses,which is responsible for the demise of large oceanic blooms formed by E.huxleyi.E.huxleyi virus(EhVs)acquired a series of enzyme-coding genes predicted to be involved in the sphingolipid biosynthesis by horizontal gene transfer between virus-host.Currently,there is limited experimental validation identifying the functions of these genes in EhV.Genetic transformation of eukaryotic cells is a powerful tool to get an insight into gene functions of the studied organisms.Serine palmitoyltransferase(SPT)catalyzes the first committed step in de novo sphingolipid biosynthetic pathway.Here,a novel vector system for the transformation of E.huxleyi was designed.It contained fragments of promoter and terminator sequences of E.huxleyi endogenic fucoxanthin chlorophyll a/c-binding protein gene“fcp”and harbored EhV-99B1 spt gene.The resultant recombinant transformation vectors pEhux-I-spt and pEhux-II were co-transferred into E.huxleyi BOF92 by electroporation.Transformants were obtained upon glufosinate-ammonium selection,and confirmed by Southern hybridization,genome PCR,qRT-PCR and Western blot screening of spt gene,which indicated that spt gene was integrated into the nuclear genome and was expressed at the mRNA and protein levels.The expression of the viral spt gene led to differences in lipid compositions analyzed using thin-layer chromatography(TLC).The results present the genetic transformation system for E.huxleyi,providing additional genetic resource with potential for exploring basic biological questions such as the virus-host interactions.展开更多
目的探讨妊娠早期血清丝氨酸蛋白酶抑制剂B1(SerpinB1)、神经调节蛋白4(NRG4)水平对妊娠期糖尿病(GDM)的预测价值。方法选取2020年12月至2022年2月在本院产检并分娩的161例GDM产妇作为GDM组,选取同期产检并分娩的152例非GDM产妇作为对...目的探讨妊娠早期血清丝氨酸蛋白酶抑制剂B1(SerpinB1)、神经调节蛋白4(NRG4)水平对妊娠期糖尿病(GDM)的预测价值。方法选取2020年12月至2022年2月在本院产检并分娩的161例GDM产妇作为GDM组,选取同期产检并分娩的152例非GDM产妇作为对照组。比较两组妊娠早期的血清SerpinB1、NRG4水平,分析其与其他血清学指标的相关性及对GDM发生和不良妊娠结局的影响及预测价值。结果GDM组的0、60、120 min口服葡萄糖耐量试验(OGTT)血糖、糖化血红蛋白(HbA1c)、甘油三酯(TG)及SerpinB1水平高于对照组,NRG4水平低于对照组(P<0.05)。妊娠早期血清SerpinB1水平与0、60、120 min OGTT血糖、HbA1c、TG呈正相关,与高密度脂蛋白胆固醇(HDL-C)呈负相关(P<0.05);妊娠早期血清NRG4水平与0、60、120 min OGTT血糖、HbA1c、低密度脂蛋白胆固醇(LDL-C)呈负相关,与HDL-C呈正相关(P<0.05)。Logistic回归分析结果显示,妊娠早期SerpinB1水平升高是GDM发生的危险因素(OR>1,P<0.05),NRG4水平升高是GDM发生的保护因素(OR<1,P<0.05)。受试者工作特征(ROC)曲线显示,血清SerpinB1、NRG4对GDM具有一定的预测价值。不良妊娠结局组的SerpinB1水平高于妊娠结局正常组,NRG4水平低于妊娠结局正常组(P<0.05)。Logistic回归分析结果显示,妊娠早期血清SerpinB1水平升高是GDM不良妊娠结局的危险因素(OR>1,P<0.05),NRG4水平升高是GDM不良妊娠结局的保护因素(OR<1,P<0.05)。ROC曲线显示,血清SerpinB1、NRG4对GDM不良妊娠结局具有一定的预测价值。结论妊娠早期血清SerpinB1、NRG4水平与GDM的发生及不良妊娠结局相关,在GDM发生及不良妊娠结局的预测中均有一定的临床价值。展开更多
Serine proteinase,purified from the hepatopancreas of Pacific white shrimp(Litopenaeus vannamei), was used to hydrolyze acid solubilized collagen(ASC)isolated from Nile tilapia(Oreochromis sp.)skin to produce angioten...Serine proteinase,purified from the hepatopancreas of Pacific white shrimp(Litopenaeus vannamei), was used to hydrolyze acid solubilized collagen(ASC)isolated from Nile tilapia(Oreochromis sp.)skin to produce angiotensin I-converting enzyme(ACE)inhibitory peptides(ACEIPs).A series of column chromatography assays were used to separate the ACEIPs.A peptide,NPARTCR,was isolated as it exhibited high ACE inhibition potential.Further digestion of this peptide by a proline specific endopeptidase(PSEP),produced a pentapeptide ARTCR with ACE inhibitory activity(IC_(50))of 77.0 pmol/L.Both NPARTCR and ARTCR inhibited ACE in a non-competitive manner.An in vivo study in rats demonstrated that ARTCR has ACE inhibitory activity via lowering systolic blood pressure in spontaneously hypertensive rats(SHRs).These results suggest that processing by-products from shrimp and tilapia are ideal raw materials for the production of serine proteinase and collagen,respectively.Serine proteinase and collagen are both ideal raw materials that can be used to derive ACE inhibitory active peptides against hypertension.展开更多
基金Supported by the National Natural Science Foundation of China(Nos.41206147,31302187)the Scientific and Technological Innovation Project Financially Supported by Qingdao National Laboratory for Marine Science and Technology(No.2015ASKJ02)
文摘Clip domain serine proteases （cSPs） and their homologs （SPHs） play an important role in various biological processes that are essential components of extracellular signaling cascades, especially in the innate immune responses of invertebrates. Here, polymorphisms of PtcSP and PtSPH from the swimming crab Portunus tritubereulatus were investigated to explore their association with resistance/ susceptibility to Vibrio alginolyticus. Polymorphic loci were identified using Clustal X, and characterized with SPSS 16.0 software, and then the significance of genotype and allele frequencies between resistant and susceptible stocks was determined by a Zz test. A total of 109 and 77 single nucleotide polymorphisms （SNPs） were identified in the genomic fragments of PtcSP and PtSPH, respectively. Notably, nearly half of PtSPH polymorphisms were found in the non-coding exon 1. Fourteen SNPs investigated were significantly associated with susceptibility/resistance to I1. alginolyticus （P〈0.05）. Among them, eight SNPs were observed in introns, and one synonymous, four non-synonymous SNPs and one ins-del were found in coding exons. In addition, five simple sequence repeats （SSRs） were detected in intron 3 of PtcSP. Although there was no statistically significant difference of allele frequencies, the SSRs showed different polymorphic alleles on the basis of the repeat number between resistant and susceptible stocks. After fiarther validation, polymorphisms investigated here might be applied to select potential molecular markers ofP. trituberculatus with resistance to I1. alginolyticus.
基金Supported by the National Natural Science Foundation of China(Nos.41576166,21707042,31771972)the Fujian Province Natural Science Foundation of China(Nos.2019J01696,2017J01447,2017J01636)。
文摘Emiliania huxleyi is the most prominent modern coccolithophore,a group of marine unicellular eukaryotes that play a critical role in ocean biogeochemistry.Coccolithoviruses are large double stranded DNA viruses,which is responsible for the demise of large oceanic blooms formed by E.huxleyi.E.huxleyi virus(EhVs)acquired a series of enzyme-coding genes predicted to be involved in the sphingolipid biosynthesis by horizontal gene transfer between virus-host.Currently,there is limited experimental validation identifying the functions of these genes in EhV.Genetic transformation of eukaryotic cells is a powerful tool to get an insight into gene functions of the studied organisms.Serine palmitoyltransferase(SPT)catalyzes the first committed step in de novo sphingolipid biosynthetic pathway.Here,a novel vector system for the transformation of E.huxleyi was designed.It contained fragments of promoter and terminator sequences of E.huxleyi endogenic fucoxanthin chlorophyll a/c-binding protein gene“fcp”and harbored EhV-99B1 spt gene.The resultant recombinant transformation vectors pEhux-I-spt and pEhux-II were co-transferred into E.huxleyi BOF92 by electroporation.Transformants were obtained upon glufosinate-ammonium selection,and confirmed by Southern hybridization,genome PCR,qRT-PCR and Western blot screening of spt gene,which indicated that spt gene was integrated into the nuclear genome and was expressed at the mRNA and protein levels.The expression of the viral spt gene led to differences in lipid compositions analyzed using thin-layer chromatography(TLC).The results present the genetic transformation system for E.huxleyi,providing additional genetic resource with potential for exploring basic biological questions such as the virus-host interactions.
文摘目的探讨妊娠早期血清丝氨酸蛋白酶抑制剂B1(SerpinB1)、神经调节蛋白4(NRG4)水平对妊娠期糖尿病(GDM)的预测价值。方法选取2020年12月至2022年2月在本院产检并分娩的161例GDM产妇作为GDM组,选取同期产检并分娩的152例非GDM产妇作为对照组。比较两组妊娠早期的血清SerpinB1、NRG4水平,分析其与其他血清学指标的相关性及对GDM发生和不良妊娠结局的影响及预测价值。结果GDM组的0、60、120 min口服葡萄糖耐量试验(OGTT)血糖、糖化血红蛋白(HbA1c)、甘油三酯(TG)及SerpinB1水平高于对照组,NRG4水平低于对照组(P<0.05)。妊娠早期血清SerpinB1水平与0、60、120 min OGTT血糖、HbA1c、TG呈正相关,与高密度脂蛋白胆固醇(HDL-C)呈负相关(P<0.05);妊娠早期血清NRG4水平与0、60、120 min OGTT血糖、HbA1c、低密度脂蛋白胆固醇(LDL-C)呈负相关,与HDL-C呈正相关(P<0.05)。Logistic回归分析结果显示,妊娠早期SerpinB1水平升高是GDM发生的危险因素(OR>1,P<0.05),NRG4水平升高是GDM发生的保护因素(OR<1,P<0.05)。受试者工作特征(ROC)曲线显示,血清SerpinB1、NRG4对GDM具有一定的预测价值。不良妊娠结局组的SerpinB1水平高于妊娠结局正常组,NRG4水平低于妊娠结局正常组(P<0.05)。Logistic回归分析结果显示,妊娠早期血清SerpinB1水平升高是GDM不良妊娠结局的危险因素(OR>1,P<0.05),NRG4水平升高是GDM不良妊娠结局的保护因素(OR<1,P<0.05)。ROC曲线显示,血清SerpinB1、NRG4对GDM不良妊娠结局具有一定的预测价值。结论妊娠早期血清SerpinB1、NRG4水平与GDM的发生及不良妊娠结局相关,在GDM发生及不良妊娠结局的预测中均有一定的临床价值。
基金This work was sponsored by the National Key R&D Program of China(2018YFD0901004)the National Natural Scientific Foundations of China(31471640,31702372).
文摘Serine proteinase,purified from the hepatopancreas of Pacific white shrimp(Litopenaeus vannamei), was used to hydrolyze acid solubilized collagen(ASC)isolated from Nile tilapia(Oreochromis sp.)skin to produce angiotensin I-converting enzyme(ACE)inhibitory peptides(ACEIPs).A series of column chromatography assays were used to separate the ACEIPs.A peptide,NPARTCR,was isolated as it exhibited high ACE inhibition potential.Further digestion of this peptide by a proline specific endopeptidase(PSEP),produced a pentapeptide ARTCR with ACE inhibitory activity(IC_(50))of 77.0 pmol/L.Both NPARTCR and ARTCR inhibited ACE in a non-competitive manner.An in vivo study in rats demonstrated that ARTCR has ACE inhibitory activity via lowering systolic blood pressure in spontaneously hypertensive rats(SHRs).These results suggest that processing by-products from shrimp and tilapia are ideal raw materials for the production of serine proteinase and collagen,respectively.Serine proteinase and collagen are both ideal raw materials that can be used to derive ACE inhibitory active peptides against hypertension.
