The mitogen-activated protein kinase kinase kinase kinases(MAP4Ks)signaling pathway plays a pivotal role in axonal regrowth and neuronal degeneration following insults.Whether targeting this pathway is beneficial to b...The mitogen-activated protein kinase kinase kinase kinases(MAP4Ks)signaling pathway plays a pivotal role in axonal regrowth and neuronal degeneration following insults.Whether targeting this pathway is beneficial to brain injury remains unclear.In this study,we showed that adeno-associated virus-delivery of the Citron homology domain of MAP4Ks effectively reduces traumatic brain injury-induced reactive gliosis,tauopathy,lesion size,and behavioral deficits.Pharmacological inhibition of MAP4Ks replicated the ameliorative effects observed with expression of the Citron homology domain.Mechanistically,the Citron homology domain acted as a dominant-negative mutant,impeding MAP4K-mediated phosphorylation of the dishevelled proteins and thereby controlling the Wnt/β-catenin pathway.These findings implicate a therapeutic potential of targeting MAP4Ks to alleviate the detrimental effects of traumatic brain injury.展开更多
The history and geographical origin of citron （Citrus medica L.） is still unknown but most of authors agree that it originated in Southeast Asia. There are different opinions about the exact period and the steps by ...The history and geographical origin of citron （Citrus medica L.） is still unknown but most of authors agree that it originated in Southeast Asia. There are different opinions about the exact period and the steps by which it was brought from its country of origin to the Mediterranean basin where it is cultivated in Corsica, Crete and in southern Italy （Calabria region）. In particulary, Citrus medica ＇Diamante＇ grows, almost exclusively, in a restricted area of the Tyrrhenian cost of the region Calabria, called ＂Riviera dei Cedri＂, a relevant local economic district. This citron has been and is, still being used by Jews for worship during the Feast of Tabernacles, a very important religious ritual, where it plays a prominent part; in fact, for this holiday of Sukkot, the use of the authentic citron that grows in Calabria region is recommended, because it is not grafted. In this paper, the fascinating history of C. medica and its spread from the centers of origin to the Mediterranean is reported with the aim to investigate the religious and cultural significance of the C. medica ＇Diamante＇, an exclusive and historical heritage of the Calabria region.展开更多
To study the effect of Green plum citron double strain fermentation filtrate(DSF)on the proliferation rate of HaCaT keratinocytes by CCK8 assay.The effect of DSF on the mRNA transcription levels of filagolin(FLG),lori...To study the effect of Green plum citron double strain fermentation filtrate(DSF)on the proliferation rate of HaCaT keratinocytes by CCK8 assay.The effect of DSF on the mRNA transcription levels of filagolin(FLG),loricrin(LOR),and occludin 1(CLD1)in HaCaT cells in sodium dodecyl sulfate(SLS)model was investigated by qPCR.The effect of DSF on the content of TNF-αin the supernatant of HaCaT cells in the model was investigated by Elisa.The results showed that DSF had no toxicity to HaCaT cells,but it could increase the mRNA transcription levels of FLG,LOR and CLD1 in HaCaT cells in the model,and reduce the content of TNF-αin the supernatant of HaCaT cells in the model.DSF had a certain effect on skin care of strengthening skin barrier.展开更多
Obesity has emerged as a growing global health concern,particularly affecting the quality of life and long-term health risks of obese individuals.Finger Citron Fruit(FCF),a functional citrus fruit,has demonstrated pot...Obesity has emerged as a growing global health concern,particularly affecting the quality of life and long-term health risks of obese individuals.Finger Citron Fruit(FCF),a functional citrus fruit,has demonstrated potential in metabolic regulation;however,its efficacy and mechanisms in mitigating postmenopausal obesity remain inadequately explored.This study systematically investigated the anti-obesity effects of FCF using an ovariectomized(OVX)mouse model and an in vitro hepatocellular steatosis model induced by oleic acid and palmitic acid(OA&PA).The results showed that FCF significantly reduced adiposity and improved metabolic rate in OVX mice.Histological and molecular analyses revealed that FCF supplementation suppressed hypertrophy and hyperplasia in both white and brown adipose tissues,downregulated hepatic lipogenic genes(Srebp-1c,Fas,Acc,Cd36),and upregulated key regulators of lipid oxidation(Ppara,Pparg).Gut microbiota sequencing indicated that FCF enhanced microbial diversity and selectively promoted the enrichment of beneficial bacteria such as such as Bacteroides,Parasutterella,Oscillibacter,and Acetatifactor,which correlated with improvements in metabolic parameters.These findings demonstrate that FCF alleviates postmenopausal obesity through dual modulation of lipid metabolic pathways and gut microbiota ecosystem.This study provides mechanistic evidence and preclinical data supporting the potential of FCF as a dietary intervention for metabolic management in postmenopausal obese populations.展开更多
Root rot is a prevalent soil-borne fungal disease in citrus.Citron C-05(Citrus medica)stands out as a germplasm within Citrus spp.due to its complete resistance to citrus canker and favorable characteristics such as s...Root rot is a prevalent soil-borne fungal disease in citrus.Citron C-05(Citrus medica)stands out as a germplasm within Citrus spp.due to its complete resistance to citrus canker and favorable characteristics such as single embryo and easy rooting.However,Citron C-05 was found to be highly susceptible to root rot during cultivation,with the specific pathogens previously unknown.In this study,four candidate fungal species were isolated from Citron C-05 roots.Sequence analysis of ITS,EF-1a,RPB1,and RPB2 identified two Fusarium solani strains,Rr-2 and Rr-4,as the candidates causing root rot in Citron C-05.Resistance tests showed these two pathogens increased root damage rate from 10.30%to 35.69%in Citron C-05,sour orange(Citrus aurantium),sweet orange(Citrus sinensis)and pummelo(Citrus grandis).F.solani exhibited the weak pathogenicity towards trifoliate orange(Poncirus trifoliata).DAB staining revealed none of reddish-brown precipitation in the four susceptible citrus germplasm after infection with F.solani,while trifoliate orange exhibited significant H2O2 accumulation.Trypan blue staining indicated increased cell death in the four susceptible citrus germplasm following infection with these two pathogens but not in trifoliate orange.These findings provide a comprehensive understanding of citrus root rot and support future research on the mechanisms of root rot resistance in citrus.展开更多
Hepatocyte growth factor-regulated tyrosine kinase substrate(Hrs)is a key component of the endosomal sorting complexes required for transport and has been demonstrated to play a regulatory role in endocytosis/exocytos...Hepatocyte growth factor-regulated tyrosine kinase substrate(Hrs)is a key component of the endosomal sorting complexes required for transport and has been demonstrated to play a regulatory role in endocytosis/exocytosis and the accumulation of internal vesicles in multivesicular bodies.Citron kinase is a Ser/The kinase that we previously reported to enhance human immunodeficiency virus type 1(HIV-1)virion production.However,the relationship between Hrs and citron kinase in HIV-1 production remains elusive.Here,we report that Hrs interacts with citron kinase via its FYVE domain.Overexpression of Hrs or the FYVE domain resulted in a significant decrease in HIV-1 virion production.Depletion of Hrs by RNA interference in HEK293T cells increased HIV-1 virion production and enhanced the activity of citron kinase.These data suggest that Hrs inhibits HIV-1 production by inhibiting citron kinase-mediated exocytosis.展开更多
The inhibitory effect of large tumor suppressor kinase(LATS1/2)on the activity of the oncoprotein yes-associated protein(YAP)is crucial to maintain tissue homeostasis.Proteomic studies have identified several new regu...The inhibitory effect of large tumor suppressor kinase(LATS1/2)on the activity of the oncoprotein yes-associated protein(YAP)is crucial to maintain tissue homeostasis.Proteomic studies have identified several new regulators of this process.Recently,citron kinase(CIT)was listed as a potential binding candidate of Hippo-related components,suggesting a new connection between CIT and the Hippo pathway.Aside from CITs role in cytokinesis,the molecular crosstalk between CIT and the Hippo pathway is largely unknown.Here,we demonstrate a role for CIT as a scaffold protein Unking LATS2 and YAP.More importantly,CIT interacts with LATS2 to directly suppress LATS2 phosphorylation at the hydrophobic motif—targeted by MST1,leading to LATS2 inactivation and YAP activation.By studying their genetic interactions,we found that Sticky,the CIT homolog in Drosophila melanogaster,functions with Warts to control Drosophila eye development.Together,our study confirms citron kinase as a novel regulator of the Hippo pathway.展开更多
Citrus pectin has been proved for its ability to bind heavy metals from industrial wastewater.However,few studies have investigated the detoxification effect of citrus pectin on cadmium-induced damage through in vivo ...Citrus pectin has been proved for its ability to bind heavy metals from industrial wastewater.However,few studies have investigated the detoxification effect of citrus pectin on cadmium-induced damage through in vivo experiments.In this study,pectin was extracted from fingered citron peel and subjected to pH-temperature modification,and successful modification of pectin’s molecular structure was confirmed by a decrease in the degree of esterification and the increase in the content of free carboxylate groups in the structure.In vitro studies suggested that the modified fingered citron peel pectin(mFCPP)showed higher cadmium removal ability compared to unmodified fingered citron peel pectin(FCPP).A 20-week study in cadmium-exposed mice suggested significant improvement in liver and kidney function with co-treatment of pectin.Co-treatment with FCPP or mFCPP ameliorated the cadmium-induced cell death and oxidative damage in the liver and kidney.Moreover,the study demonstrated that both FCPP and mFCPP were impermeable through the intestinal wall,suggesting their binding with the cadmium ions in the gastrointestinal tract,thus preventing cadmium ions absorption and entry into the systemic circulation.展开更多
In this study,the structure-activity relationship of finger citron polysaccharides(FCPs)and their phosphorylated derivatives(P-FCPs)was investigated.Three FCPs(FCP20,FCP40 and FCP60)were successively fractionated by e...In this study,the structure-activity relationship of finger citron polysaccharides(FCPs)and their phosphorylated derivatives(P-FCPs)was investigated.Three FCPs(FCP20,FCP40 and FCP60)were successively fractionated by ethanol,and were phosphorylated modification to obtain P-FCPs(P-FCP20,P-FCP40 and P-FCP60).FCPs and P-FCPs are acidic heteropolysaccharides composed of mannose,rhamnose,glucose,galactose,arabinose and galacturonic acid,and exhibited different content of neutral sugar,uronic acid,proteins,polyphenols and sulfates.FTIR and NMR analyses determined the structures of FCPs and P-FCPs,indicating that the phosphorylation modification was successful.P-FCPs possessed the smaller molecular weight distribution and particle size,with significant changes in microstructure than FCPs.Additionally,P-FCPs showed significantly increased antioxidant and α-amylase inhibitory activities compared to FCPs.The inhibitory mechanisms of FCP40 and P-FCP40 on α-amylase was a mixed-type manner and dynamic fluorescence quenching.The results demonstrate that phosphorylation modification can alter the structural features and bioactivities of FCPs,providing an approach for the further development of finger citron in the field of medicinal and functional foods.展开更多
Citrus canker,caused by Xanthomonas citri subsp.citri(Xcc),is a globally quarantine disease infecting nearly all Citrus cultivars.Citron C-05 has been identified with complete and active resistance to Xcc.However,the ...Citrus canker,caused by Xanthomonas citri subsp.citri(Xcc),is a globally quarantine disease infecting nearly all Citrus cultivars.Citron C-05 has been identified with complete and active resistance to Xcc.However,the mechanism underlying Citron C-05's resistance to Xcc remains elusive.We identified a gene cluster on chromosome 8 of the citrus genome comprising five pathogenesis-related 4-like genes.PR4A was upregulated in Citron C-05 leaves under Xcc infection,exhibiting the highest expression among these PR4-like genes.In addition,PR4A expression was higher in leaves of disease-resistant genotypes than susceptible genotypes under Xcc invasion.Bimolecular fluorescence complementation(BiFC)and Split-Luc assays indicated that CmWRKY75,a positive regulator of PR4A,interacted with pthA4 and upregulated expression of PR4A in Citron C-05 leaves.Regulatory function for the expression of CmPR4A was localized to a 516-nucleotide region upstream of the translational start site,which was designated Pro CmPR4A-P516.Transient overexpression of CmPR4A improved resistance to Xcc in sweet orange,and three transgenic lines of OE-CmPR4A exhibited resistance to Pseudomonas syringae pv.tomato DC3000(Pst DC3000)in Arabidopsis.Furthermore,CmSMU2 was identified through yeast two-hybrid library using CmPR4A as bait,Bi FC and Split-Luc assays further verified their interaction.Transient overexpression of CmSMU2 in sweet orange increased resistance to Xcc.Co-expression of CmSMU2 and CmPR4A enhanced accumulation of reactive oxygen species compared to CmSMU2 or CmPR4A,indicating that they may synergistically enhance resistance to Xcc in citrus.These findings lay the groundwork for a theoretical analysis of the mechanism underlying the resistance of Citron C-05 against citrus canker.展开更多
基金supported by the TARCC,Welch Foundation Award(I-1724)the Decherd Foundationthe Pape Adams Foundation,NIH grants NS092616,NS127375,NS117065,NS111776。
文摘The mitogen-activated protein kinase kinase kinase kinases(MAP4Ks)signaling pathway plays a pivotal role in axonal regrowth and neuronal degeneration following insults.Whether targeting this pathway is beneficial to brain injury remains unclear.In this study,we showed that adeno-associated virus-delivery of the Citron homology domain of MAP4Ks effectively reduces traumatic brain injury-induced reactive gliosis,tauopathy,lesion size,and behavioral deficits.Pharmacological inhibition of MAP4Ks replicated the ameliorative effects observed with expression of the Citron homology domain.Mechanistically,the Citron homology domain acted as a dominant-negative mutant,impeding MAP4K-mediated phosphorylation of the dishevelled proteins and thereby controlling the Wnt/β-catenin pathway.These findings implicate a therapeutic potential of targeting MAP4Ks to alleviate the detrimental effects of traumatic brain injury.
文摘The history and geographical origin of citron （Citrus medica L.） is still unknown but most of authors agree that it originated in Southeast Asia. There are different opinions about the exact period and the steps by which it was brought from its country of origin to the Mediterranean basin where it is cultivated in Corsica, Crete and in southern Italy （Calabria region）. In particulary, Citrus medica ＇Diamante＇ grows, almost exclusively, in a restricted area of the Tyrrhenian cost of the region Calabria, called ＂Riviera dei Cedri＂, a relevant local economic district. This citron has been and is, still being used by Jews for worship during the Feast of Tabernacles, a very important religious ritual, where it plays a prominent part; in fact, for this holiday of Sukkot, the use of the authentic citron that grows in Calabria region is recommended, because it is not grafted. In this paper, the fascinating history of C. medica and its spread from the centers of origin to the Mediterranean is reported with the aim to investigate the religious and cultural significance of the C. medica ＇Diamante＇, an exclusive and historical heritage of the Calabria region.
文摘To study the effect of Green plum citron double strain fermentation filtrate(DSF)on the proliferation rate of HaCaT keratinocytes by CCK8 assay.The effect of DSF on the mRNA transcription levels of filagolin(FLG),loricrin(LOR),and occludin 1(CLD1)in HaCaT cells in sodium dodecyl sulfate(SLS)model was investigated by qPCR.The effect of DSF on the content of TNF-αin the supernatant of HaCaT cells in the model was investigated by Elisa.The results showed that DSF had no toxicity to HaCaT cells,but it could increase the mRNA transcription levels of FLG,LOR and CLD1 in HaCaT cells in the model,and reduce the content of TNF-αin the supernatant of HaCaT cells in the model.DSF had a certain effect on skin care of strengthening skin barrier.
基金supported by grants from the project of Hainan Medical University(RZ2300007534)the Hainan Provincial Natural-Science Foundation of China(824RC522)the National Natural Science Foundation of China(82360882,82304826).
文摘Obesity has emerged as a growing global health concern,particularly affecting the quality of life and long-term health risks of obese individuals.Finger Citron Fruit(FCF),a functional citrus fruit,has demonstrated potential in metabolic regulation;however,its efficacy and mechanisms in mitigating postmenopausal obesity remain inadequately explored.This study systematically investigated the anti-obesity effects of FCF using an ovariectomized(OVX)mouse model and an in vitro hepatocellular steatosis model induced by oleic acid and palmitic acid(OA&PA).The results showed that FCF significantly reduced adiposity and improved metabolic rate in OVX mice.Histological and molecular analyses revealed that FCF supplementation suppressed hypertrophy and hyperplasia in both white and brown adipose tissues,downregulated hepatic lipogenic genes(Srebp-1c,Fas,Acc,Cd36),and upregulated key regulators of lipid oxidation(Ppara,Pparg).Gut microbiota sequencing indicated that FCF enhanced microbial diversity and selectively promoted the enrichment of beneficial bacteria such as such as Bacteroides,Parasutterella,Oscillibacter,and Acetatifactor,which correlated with improvements in metabolic parameters.These findings demonstrate that FCF alleviates postmenopausal obesity through dual modulation of lipid metabolic pathways and gut microbiota ecosystem.This study provides mechanistic evidence and preclinical data supporting the potential of FCF as a dietary intervention for metabolic management in postmenopausal obese populations.
基金supported by Joint Funds of the National Natural Science Foundation of China(Grant No.U21A20228).
文摘Root rot is a prevalent soil-borne fungal disease in citrus.Citron C-05(Citrus medica)stands out as a germplasm within Citrus spp.due to its complete resistance to citrus canker and favorable characteristics such as single embryo and easy rooting.However,Citron C-05 was found to be highly susceptible to root rot during cultivation,with the specific pathogens previously unknown.In this study,four candidate fungal species were isolated from Citron C-05 roots.Sequence analysis of ITS,EF-1a,RPB1,and RPB2 identified two Fusarium solani strains,Rr-2 and Rr-4,as the candidates causing root rot in Citron C-05.Resistance tests showed these two pathogens increased root damage rate from 10.30%to 35.69%in Citron C-05,sour orange(Citrus aurantium),sweet orange(Citrus sinensis)and pummelo(Citrus grandis).F.solani exhibited the weak pathogenicity towards trifoliate orange(Poncirus trifoliata).DAB staining revealed none of reddish-brown precipitation in the four susceptible citrus germplasm after infection with F.solani,while trifoliate orange exhibited significant H2O2 accumulation.Trypan blue staining indicated increased cell death in the four susceptible citrus germplasm following infection with these two pathogens but not in trifoliate orange.These findings provide a comprehensive understanding of citrus root rot and support future research on the mechanisms of root rot resistance in citrus.
基金in part supported by grants to Guangxia Gao from the National Natural Science Foundation of China(Grant No.81030030)the Ministry of Science and Technology of China(No.2009zx09501-029).
文摘Hepatocyte growth factor-regulated tyrosine kinase substrate(Hrs)is a key component of the endosomal sorting complexes required for transport and has been demonstrated to play a regulatory role in endocytosis/exocytosis and the accumulation of internal vesicles in multivesicular bodies.Citron kinase is a Ser/The kinase that we previously reported to enhance human immunodeficiency virus type 1(HIV-1)virion production.However,the relationship between Hrs and citron kinase in HIV-1 production remains elusive.Here,we report that Hrs interacts with citron kinase via its FYVE domain.Overexpression of Hrs or the FYVE domain resulted in a significant decrease in HIV-1 virion production.Depletion of Hrs by RNA interference in HEK293T cells increased HIV-1 virion production and enhanced the activity of citron kinase.These data suggest that Hrs inhibits HIV-1 production by inhibiting citron kinase-mediated exocytosis.
文摘The inhibitory effect of large tumor suppressor kinase(LATS1/2)on the activity of the oncoprotein yes-associated protein(YAP)is crucial to maintain tissue homeostasis.Proteomic studies have identified several new regulators of this process.Recently,citron kinase(CIT)was listed as a potential binding candidate of Hippo-related components,suggesting a new connection between CIT and the Hippo pathway.Aside from CITs role in cytokinesis,the molecular crosstalk between CIT and the Hippo pathway is largely unknown.Here,we demonstrate a role for CIT as a scaffold protein Unking LATS2 and YAP.More importantly,CIT interacts with LATS2 to directly suppress LATS2 phosphorylation at the hydrophobic motif—targeted by MST1,leading to LATS2 inactivation and YAP activation.By studying their genetic interactions,we found that Sticky,the CIT homolog in Drosophila melanogaster,functions with Warts to control Drosophila eye development.Together,our study confirms citron kinase as a novel regulator of the Hippo pathway.
基金supported by the Natural Science Foundation of Zhejiang Province(No.Y23C200019)the Special Fund for Basic Operating Expenses of Colleges and Universities Affiliated to Zhejiang Province(No.3090JYN9921001G-017).
文摘Citrus pectin has been proved for its ability to bind heavy metals from industrial wastewater.However,few studies have investigated the detoxification effect of citrus pectin on cadmium-induced damage through in vivo experiments.In this study,pectin was extracted from fingered citron peel and subjected to pH-temperature modification,and successful modification of pectin’s molecular structure was confirmed by a decrease in the degree of esterification and the increase in the content of free carboxylate groups in the structure.In vitro studies suggested that the modified fingered citron peel pectin(mFCPP)showed higher cadmium removal ability compared to unmodified fingered citron peel pectin(FCPP).A 20-week study in cadmium-exposed mice suggested significant improvement in liver and kidney function with co-treatment of pectin.Co-treatment with FCPP or mFCPP ameliorated the cadmium-induced cell death and oxidative damage in the liver and kidney.Moreover,the study demonstrated that both FCPP and mFCPP were impermeable through the intestinal wall,suggesting their binding with the cadmium ions in the gastrointestinal tract,thus preventing cadmium ions absorption and entry into the systemic circulation.
基金financial support from the Guangdong Provincial Medical Research Foundation(A2024095)Scientific Research Project of Guangdong Provincial Bureau of Traditional Chinese Medicine(20231216).
文摘In this study,the structure-activity relationship of finger citron polysaccharides(FCPs)and their phosphorylated derivatives(P-FCPs)was investigated.Three FCPs(FCP20,FCP40 and FCP60)were successively fractionated by ethanol,and were phosphorylated modification to obtain P-FCPs(P-FCP20,P-FCP40 and P-FCP60).FCPs and P-FCPs are acidic heteropolysaccharides composed of mannose,rhamnose,glucose,galactose,arabinose and galacturonic acid,and exhibited different content of neutral sugar,uronic acid,proteins,polyphenols and sulfates.FTIR and NMR analyses determined the structures of FCPs and P-FCPs,indicating that the phosphorylation modification was successful.P-FCPs possessed the smaller molecular weight distribution and particle size,with significant changes in microstructure than FCPs.Additionally,P-FCPs showed significantly increased antioxidant and α-amylase inhibitory activities compared to FCPs.The inhibitory mechanisms of FCP40 and P-FCP40 on α-amylase was a mixed-type manner and dynamic fluorescence quenching.The results demonstrate that phosphorylation modification can alter the structural features and bioactivities of FCPs,providing an approach for the further development of finger citron in the field of medicinal and functional foods.
基金supported by the National Natural Science Foundation of China(32402500 and 31741110)the Scientific Research Fund of Hunan Provincial Education Department,China(21B0220)。
文摘Citrus canker,caused by Xanthomonas citri subsp.citri(Xcc),is a globally quarantine disease infecting nearly all Citrus cultivars.Citron C-05 has been identified with complete and active resistance to Xcc.However,the mechanism underlying Citron C-05's resistance to Xcc remains elusive.We identified a gene cluster on chromosome 8 of the citrus genome comprising five pathogenesis-related 4-like genes.PR4A was upregulated in Citron C-05 leaves under Xcc infection,exhibiting the highest expression among these PR4-like genes.In addition,PR4A expression was higher in leaves of disease-resistant genotypes than susceptible genotypes under Xcc invasion.Bimolecular fluorescence complementation(BiFC)and Split-Luc assays indicated that CmWRKY75,a positive regulator of PR4A,interacted with pthA4 and upregulated expression of PR4A in Citron C-05 leaves.Regulatory function for the expression of CmPR4A was localized to a 516-nucleotide region upstream of the translational start site,which was designated Pro CmPR4A-P516.Transient overexpression of CmPR4A improved resistance to Xcc in sweet orange,and three transgenic lines of OE-CmPR4A exhibited resistance to Pseudomonas syringae pv.tomato DC3000(Pst DC3000)in Arabidopsis.Furthermore,CmSMU2 was identified through yeast two-hybrid library using CmPR4A as bait,Bi FC and Split-Luc assays further verified their interaction.Transient overexpression of CmSMU2 in sweet orange increased resistance to Xcc.Co-expression of CmSMU2 and CmPR4A enhanced accumulation of reactive oxygen species compared to CmSMU2 or CmPR4A,indicating that they may synergistically enhance resistance to Xcc in citrus.These findings lay the groundwork for a theoretical analysis of the mechanism underlying the resistance of Citron C-05 against citrus canker.
