Dear Editor,The Cay2.1 channel,also known as the P/Q-type Ca^(2+) channel,is a particular type of voltage-gated Ca^(2+) channel primarily expressed on the presynaptic membrane in the brain[1].It serves as an essential...Dear Editor,The Cay2.1 channel,also known as the P/Q-type Ca^(2+) channel,is a particular type of voltage-gated Ca^(2+) channel primarily expressed on the presynaptic membrane in the brain[1].It serves as an essential part of the precisely orchestrated neurotransmitter release machinery.展开更多
Nonhuman primates are increasingly being used as animal models in neuroscience research.However,efficient neuronal tracing techniques for labeling motor neurons and primary sensory afferents in the monkey spinal cord ...Nonhuman primates are increasingly being used as animal models in neuroscience research.However,efficient neuronal tracing techniques for labeling motor neurons and primary sensory afferents in the monkey spinal cord are lacking.Here,by injecting the cholera toxin B subunit into the sciatic nerve of a rhesus monkey,we successfully labeled the motor neurons and primary sensory afferents in the lumbar and sacralspinal cord.Labeled alpha motor neurons were located in lamina IX of the L6–S1 segments,which innervate both flexors and extensors.The labeled primary sensory afferents were mainly myelinated Aβfibers that terminated mostly in laminae I and II of the L4–L7 segments.Together with the labeled proprioceptive afferents,the primary sensory afferents formed excitatory synapses with multiple types of spinal neurons.In summary,our methods successfully traced neuronal connections in the monkey spinal cord and can be used in spinal cord studies when nonhuman primates are used.展开更多
Premenstrual dysphoric disorder(PMDD),a subtype of premenstrual syndrome(PMS),involves physical and emotional symptoms that impact patients'daily lives and productivity.A reliable,side-effect-free clinical interve...Premenstrual dysphoric disorder(PMDD),a subtype of premenstrual syndrome(PMS),involves physical and emotional symptoms that impact patients'daily lives and productivity.A reliable,side-effect-free clinical intervention is needed.Shuyu capsule is an effective traditional Chinese medicine preparation for PMDD used in the clinics,but its therapeutic mechanism remains unclear.Previous research has suggested that theγ-aminobutyric acidergic(GABAergic)system in the periaqueductal gray(PAG)may play a role in treating PMDD with traditional Chinese medicine,but there is a lack of functional verification.This study aims to reveal the potential mechanism of the Shuyu capsule in treating PMDD.The study employed an experimental design using female C57BL/6J and Vgat-Cre mice to assess the effects of Shuyu capsules on PMDD,with a focus on the GABAergic system in the dorsal PAG(dPAG).Assessments were conducted using the forced swimming test(FST)to gauge depression-like behaviors and western blot(WB)and immunofluorescence(IF)to measure the numbers of active GABAergic neurons and theγ-aminobutyric acid type A receptor(GABA,R)δsubunit(GABRD)expression.Chemogenetic techniques and adeno-associated virus were specifically used to activate GABAergic neurons and knock down the expression of subunits,respectively,providing insights into the neurobiological mechanisms underpinning the therapeutic effects of Shuyu capsules in treating PMDD.After being stressed by FST,the immobility duration of PMDD mice in the late dioestrus(LD)phase decreased after the Shuyu capsule intervention,implying that it can improve the estrous cycle-dependent depression-like phenotype in PMDD mice.Additionally,the application of Shuyu capsule can downregulate the expression of GABRD and reverse the downtrend of activated GABAergic neurons in the dPAG of PMDD model mice.We also found that single-target manipulation was enough to improve the depressionlike behavior of PMDD model mice.Transgenic mice with GABRD knockout were established,and their behaviors were tested,revealing changes in their exploratory behaviors,indicating that the GABRD may be closely related to anxiety disorders.Shuyu capsule plays an anti-PMDD role by activating GABAergic neurons and downregulating the expression of GABRD in the dPAG.This provides a theoretical basis for the clinical treatment of PMDD with traditional Chinese medicine and promotes the development of drugs for treating PMDD.展开更多
BACKGROUND In recent years,many studies have shown that proteasome 26S subunit non-ATPase 6(PSMD6)plays an important role in the occurrence and development of malignant tumours.Unfortunately,there are no reports on th...BACKGROUND In recent years,many studies have shown that proteasome 26S subunit non-ATPase 6(PSMD6)plays an important role in the occurrence and development of malignant tumours.Unfortunately,there are no reports on the evaluation of the potential role of PSMD6 in hepatocellular carcinoma(HCC).AIM To comprehensively evaluate the overexpression pattern and clinical significance of PSMD6 in HCC tissues.METHODS This study integrated PSMD6 mRNA expression profiles from 4672 HCC and 3667 non-HCC tissues,along with immunohistochemical scores from 383 HCC and adjacent tissues,to assess PSMD6 overexpression in HCC.Clustered regularly interspaced short palindromic repeats knockout technology evaluated PSMD6’s essential role in HCC cell growth.Functional enrichment analysis explored the molecular mechanism of PSMD6 abnormalities in HCC.Drug sensitivity analysis and molecular docking analysed the effect of abnormal expression of PSMD6 on the drug sensitivity of HCC cells.RESULTS The results of 41 external and two internal datasets showed that PSMD6 mRNA(SMD=0.26,95%CI:0.09-0.42,P<0.05)and protein(SMD=2.85,95%CI:1.19-4.50,P<0.05)were significantly overexpressed in HCC tissues.The integrated analysis results showed that PSMD6 had a significant overexpression pattern in HCC tissues(SMD=0.40,95%CI:0.15-0.66,P<0.05).PSMD6 knockout inhibited HCC cell growth(chronos scores<-1).Functional enrichment implicated ribosome biogenesis and RNA splicing.Significant enrichment of signalling pathways such as RNA degradation,ribosomes,and chemical carcinogenesis—reactive oxygen species.Drug sensitivity analysis and a molecular docking model showed that high expression of PSMD6 was associated with the tolerance of HCC cells to drugs such as ML323,sepantronium bromide,and GDC0810.Overexpressed PSMD6 effectively distinguished HCC tissues(AUC=0.75,95%CI:0.71-0.79).CONCLUSION This study was the first to discover that PSMD6 was overexpressed in HCC tissues.PSMD6 is essential for the growth of HCC cells and may be involved in ribosome biogenesis and RNA splicing.展开更多
Improving protein quality and grain yield traits coordinately is an important goal for crop breeding.To date,many protein-quality or grain-yield regulation genes have been identified.However,the genetic strategies int...Improving protein quality and grain yield traits coordinately is an important goal for crop breeding.To date,many protein-quality or grain-yield regulation genes have been identified.However,the genetic strategies integrating these genes in good-protein-quality and high-yield crop breeding practice are far from established.Here,we characterized the functions of the MADS domain-containing protein Zm MADS8 and Zea mays G protein gamma subunit 1(Zm GG1)in regulating protein quality and grain yield of maize.Zm MADS8 positively regulates zein protein accumulation and negatively regulates nonzein protein and lysine levels in kernels by interacting with Zm MADS47 to promote the transcriptional activation of Opaque2.Additionally,Zm MADS8 regulates starch content of kernels by targeting genes involved in starch biosynthesis.Zm GG1,a putative interactor of Zm MADS8,negatively regulates kernel number with a trade-off effect on kernel starch accumulation.The mads8;zmgg1 double mutant improved protein quality by attenuating zein biosynthesis and increasing essential lysine level,and increased grain yield by increasing kernel number,compensating for decreased starch biosynthesis.Our findings revealed the biological function of Zm MADS8 and Zm GG1 in regulating protein quality and yield related traits and suggested a genetic strategy by direct editing of Zm MADS8 and Zm GG1 to improve grain yield and protein quality simultaneously.展开更多
BACKGROUND As a member of the chaperonin-containing tailless complex polypeptide 1(TCP1)complex,which plays a pivotal role in ensuring the accurate folding of numerous proteins,chaperonin-containing TCP1 subunit 6A(CC...BACKGROUND As a member of the chaperonin-containing tailless complex polypeptide 1(TCP1)complex,which plays a pivotal role in ensuring the accurate folding of numerous proteins,chaperonin-containing TCP1 subunit 6A(CCT6A)participates in various physiological and pathological processes.However,its effects on cell death and cancer therapy and the underlying mechanisms need further exploration in colorectal cancer(CRC)cells.AIM To explore the effects of CCT6A on cell death and cancer therapy and the underlying mechanisms in CRC.METHODS Cell proliferation was evaluated using the MTS assay,EdU staining,and colony growth assays.The expression of CCT6A was monitored by immunoblotting and quantitative PCR.CCT6A was knocked out by CRISPR-Cas9,and overexpressed by transfecting plasmids.Autophagy was examined by immunoblotting and the mCherry-GFP-LC3 assay.To monitor apoptosis and necroptosis,immunoblotting,co-immunoprecipitation,and flow cytometry were employed.RESULTS Cisplatin(DDP)exerted cytotoxic effects on CRC cells while simultaneously downregulating the expression of CCT6A.Depletion of CCT6A amplified the cytotoxic effects of DDP,whereas overexpression of CCT6A attenuated these adverse effects.CCT6A suppressed autophagy,apoptosis,and necroptosis under both basal and DDP-treated conditions.Autophagy inhibitors significantly enhanced the cytotoxic effects of DDP,whereas a necroptosis inhibitor partially reversed the cell viability loss induced by DDP.Furthermore,inhibiting autophagy enhanced both apoptosis and necroptosis induced by DDP.CONCLUSION CCT6A negatively modulates autophagy,apoptosis,and necroptosis,and CCT6A confers resistance to DDP therapy in CRC,suggesting its potential as a therapeutic target.展开更多
BACKGROUND Hepatocellular carcinoma(HCC)is a leading cause of cancer-related mortality worldwide,with hepatitis B virus(HBV)infection serving as a significant etiological factor in endemic regions.Alpha-fetoprotein(AF...BACKGROUND Hepatocellular carcinoma(HCC)is a leading cause of cancer-related mortality worldwide,with hepatitis B virus(HBV)infection serving as a significant etiological factor in endemic regions.Alpha-fetoprotein(AFP),the most commonly used biomarker,has limited sensitivity,particularly in AFP-negative HCC.Recent studies have identified origin recognition complex subunit 1(ORC1)and extra spindle pole bodies-like 1(ESPL1)as promising serum biomarkers,both linked to HBV DNA integration,a mechanism known to drive hepatocarcinogenesis.AIM To assess serum ORC1’s diagnostic value for HBV-HCC and its link to S gene integration.METHODS In this case-control study,479 HBV-infected patients were enrolled,including 20 hepatitis B,154 with HBV-related cirrhosis,and 96 with HBV-HCC.The control group comprised 73 individuals:29 with non-HBV-HCC and 44 healthy participants.Serum ORC1 and ESPL1 were measured by enzyme-linked immunosorbent assay.HBV integration sites were identified via whole-genome sequencing.Diagnostic performance was assessed using receiver operating characteristic analysis,including in AFP-negative patients.RESULTS HBV integration near the ORC1 locus(chromosome 1p32.3)was detected in 71.4%of HBV-HCC tissues.Serum ORC1 levels were significantly higher in HBV-infected patients than in non-HBV-infected controls(980.11 ng/L vs 746.82 ng/L,P<0.05)and in HBV-HCC compared with non-HBV-HCC(1077.07 ng/L vs 749.54 ng/L,P<0.05).Serum ORC1 and ESPL1 were elevated in HBV-HCC regardless of AFP status,and detected 64.8%and 73.2%of AFP-negative cases,respectively.The combined panel of ORC1[Area under receiver operating characteristic curve(AUC)=0.587],ESPL1(AUC=0.776),and AFP(AUC=0.844)achieved an AUC of 0.887,significantly higher than any single marker(P<0.05),with a sensitivity of 84.44%,specificity of 84.19%,and a negative predictive value of 94.91%.CONCLUSION Serum ORC1,driven by HBV integration,is a promising biomarker especially for AFP-negative HBV-HCC.Its combination with ESPL1 and AFP significantly improves early detection.展开更多
BACKGROUND Laryngeal squamous cell carcinoma(LSCC)is a prevalent head and neck malignancy with suboptimal survival rates due to late detection and therapeutic resistance.AIM To investigate chaperonin-containing TCP1 s...BACKGROUND Laryngeal squamous cell carcinoma(LSCC)is a prevalent head and neck malignancy with suboptimal survival rates due to late detection and therapeutic resistance.AIM To investigate chaperonin-containing TCP1 subunit 3(CCT3)expression and its clinical implications,and its effects on LSCC cell growth.METHODS Systematic data on CCT3 mRNA expression were collected from biomedical databases,and integrated further based on the standardized mean difference and the summary receiver operating characteristic curve.Single-cell RNA-seq data were mined to validate the expression level of CCT3 mRNA.In-house immunohistochemistry was performed to explore the CCT3 protein levels of clinical LSCC samples and their relationship with clinical parameters.The growth function of LSCC cell was analyzed using CRISPR knockout screening.CCT3-related signaling pathway analyses were conducted using gene set enrichment analysis.Protein-protein interaction network construction was performed to identify hub genes.RESULTS CCT3 mRNA was significantly overexpressed in 269 LSCC tissues cases across multiple independent datasets(standardized mean difference=32,area under the curve=0.93);At the translational level,the in-house immunohistochemical analysis further demonstrated the consistent upregulation of CCT3 protein in 88 cases of LSCC samples(58 non-LSCC samples vs 30 LSCC samples,P=1.4e^(-14)).Analysis of clinical parameters showed no significant differences among subgroup.Functional characterization with clustered regularly interspaced short palindromic repeats--mediated gene knockout revealed that depletion of CCT3 potently suppressed LSCC cell viability in vitro.Gene set enrichment analysis indicated that CCT3 was markedly associated with several key oncogenic pathways,including extracellular matrix receptor interaction and cell cycle regulation pathways.CONCLUSION CCT3 upregulation in LSCC may influence cellular growth by regulating related pathways,indicating its potential as a biomarker and therapeutic target for LSCC.展开更多
A deficiency ofγδT cells has been described in Crohn's disease(CD).AIM To analyze the gene expression of interleukin 7(IL-7)and its receptors in the tissues of patients with CD.METHODS We studied the peripheral ...A deficiency ofγδT cells has been described in Crohn's disease(CD).AIM To analyze the gene expression of interleukin 7(IL-7)and its receptors in the tissues of patients with CD.METHODS We studied the peripheral blood of 80 patients with CD,comparing them with a group of 80 healthy subjects.The number and apoptosis ofαβandγδT cells in peripheral blood and the proportion ofαβandγδT cells in the intestinal tissues of patients with CD(n=25)were studied.The gene and protein expression of IL-7,IL-2 receptor subunitγ[cluster of differentiation 132(CD132)],receptorα(CD127),and caspase-3 in tissues was analyzed by quantitative PCR.Serum IL-7 levels were also analyzed.RESULTS In patients with CD,a decreased number ofγδT cells and an increase in the apoptosis of CD56+αβandγδT cells in peripheral blood was observed(P<0.0001 and P<0.01)respectively,and there was an inverse correlation among T subsets and their apoptosis.In addition,IL-7 gene expression and IL-7 protein in the tissues of these patients were increased.The titers of caspase-3 in tissues were low vs control group(P>0.01).The percentage of CD8+γδT cells decreased in tissues(P<0.01),and was directly related to IL-7 levels in peripheral blood.The expression of IL-2 receptor subunitγ(CD132)was greatly decreased in the tissues of patients with CD(P<0.05).CONCLUSION There may be a cause-effect relationship between the lower gene expression of the IL-2 receptor subunitγ(CD132)in tissues of patients with CD andγδT cells immunodeficiency.展开更多
Mosquito-borne flaviviruses,such as Zika virus(ZIKV)and dengue virus(DENV),cause diverse severe clinical manifestations including fever,rash,hepatitis,arthralgia,and congenital anomalies.Here,we identified a host fact...Mosquito-borne flaviviruses,such as Zika virus(ZIKV)and dengue virus(DENV),cause diverse severe clinical manifestations including fever,rash,hepatitis,arthralgia,and congenital anomalies.Here,we identified a host factor,the adaptor protein complex 1 gamma 1 subunit(AP1G1),which plays an important role in both ZIKV and dengue virus 2(DENV2)infections.We explored the role of AP1G1 in ZIKV and DENV2 infections using CRISPR/Cas9 gene editing technology and RNA interference(RNAi)techniques.Knockout or silencing of AP1G1 decreases the replication of ZIKV and DENV2 in multiple human cell lines.Intriguingly,depletion of AP1G1 results in a significant reduction in ZIKV at an early stage,but decreases DENV2 replication levels during the late stage,suggesting that AP1G1 plays distinct roles in the infection by ZIKV and DENV2.Furthermore,we determined that AP1G1 mediates ZIKV-endosomal membrane fusion through inhibitor experiments and fluorescence labeling assays.Mechanistically,we found that AP1G1 exerts its pro-viral effect through binding to the ZIKV envelope glycoprotein(E protein).This interaction promotes the fusion of viral and endosomal membranes,during which the ZIKV genomic RNAs are released from the endosome into the cytoplasm,a process that facilitates viral replication.However,for DENV2 infection,AP1G1 primarily affects its viral RNA replication stage,rather than the fusion of virus-endosomal membrane.Taken together,our work demonstrates that AP1G1 plays a pro-viral role in both ZIKV and DENV2 infections via distinct mechanisms,highlighting its potential as a therapeutic target for antiviral strategies.展开更多
Crohn’s disease(CD)is a chronic inflammatory disorder characterized by dysregulated immune responses and significant disruption of intestinal immunity.A recent case-control study by Andreu-Ballester et al revealed de...Crohn’s disease(CD)is a chronic inflammatory disorder characterized by dysregulated immune responses and significant disruption of intestinal immunity.A recent case-control study by Andreu-Ballester et al revealed decreased expression of interleukin(IL)-2 receptor subunitγ(CD132)in CD tissues,a finding that has profound implications for understanding immune dysregulation in CD.CD132,an essential component of the IL-7/IL-2 signaling axis,is critical forγδT cell survival and function,which are pivotal for maintaining gut integrity and modulating inflammation.Here,we propose that reduced CD132 expression represents a key mechanism underlyingγδT cell deficiencies in CD,contributing to impaired immune surveillance and exacerbated inflammation.This hypothesis integrates emerging evidence from cytokine signaling and immunopathology in CD,offering new insights into its pathogenesis.These findings highlight the therapeutic potential of targeting the IL-7/IL-2 axis to restore immune homeostasis in CD,presenting a novel avenue for future research and intervention.展开更多
Nicotinic acetylcholine receptors (nAChRs) play a significant role in excitatory synaptic transmission in insects and are the target for chloronicotinyl and nereistoxin insecticides.In recent years,Chilo suppressalis,...Nicotinic acetylcholine receptors (nAChRs) play a significant role in excitatory synaptic transmission in insects and are the target for chloronicotinyl and nereistoxin insecticides.In recent years,Chilo suppressalis,an economically important pest of rice,developed high resistance against monosultap,a nereistoxin insecticide acting on nAChR.In order to reveal the hypothesized target insensitive mechanism,studies on the molecular property of nAChR from Chilo suppressalis are required.In this study,the full length cDNA of nAChR α subunit from this pest was cloned by RT-PCR.Sequence analysis shows that it is a novel nAChR α subunit,which was named as Cs α 1(Genbank accession No.AF418987).It contains 1?997?bp nucleotides and involves an open reading frame (ORF) encoding a mature protein of 509 amino acids excluding a signal peptide of 24 amino acids.The deduced amino acid sequence was 52%-94% identical to the reported insect nAChR genes.展开更多
A vacuolar ATPase (V-ATPase.) B subunit gene has been cloned and characterized front a phosphorus starvation induced rice root subtractive cDNA library by suppression subtractive hybridization (SSH) method and RT-PCR ...A vacuolar ATPase (V-ATPase.) B subunit gene has been cloned and characterized front a phosphorus starvation induced rice root subtractive cDNA library by suppression subtractive hybridization (SSH) method and RT-PCR amplification. This gene encodes a polypeptide of 487 amino acid residues, containing a conservative ATP binding site and with a molecular weight of 54.06 kD and an isoelectric point of 4.99, southern analysis of the. genomic DNA indicates that V-ATPase B subunit is encoded by a single gene in rice genome. The amino acid homologies of V-ATPase B subunits among different organisms range from 76% to 97% and reveals that the evolution of V-ATPase B subunit is accompanied with the biological evolution. Expression pattern analysis indicated that the maximal expression of V-ATPase B subunit gene occurred at an early stage (6 - 12 h) after phosphorus starvation in roots, and lately stage (24 - 48 It) in leaves. Under phosphorus deficiency, the up-regulated expression of V-ATPase gene was presumed to strengthen the proton transport and provide the required energy to maintain an electrochemical gradient across the tonoplast to facilitate Phosphorus transport.展开更多
[Objective]The α subunit gene of phycobiliprotein from Spirulina maxima was studied in order to provide a basis for the subsequent study of phycobiliprotein.[Method] Amino acids composition,signal peptides,hydrophobi...[Objective]The α subunit gene of phycobiliprotein from Spirulina maxima was studied in order to provide a basis for the subsequent study of phycobiliprotein.[Method] Amino acids composition,signal peptides,hydrophobicity/hydrophilicity and trails-membrane topological structure of α subunit gene of phycobiliprotein from Spirulina maxima which registered in GenBank（GenBank AF441177） were analyzed and predicted by the tools of bioinformatic analysis.Meanwhile,phylogenetic tree was constructed based on α subunit gene of phycobiliprotein from Spirulina maxima,and its molecular evolution was also analyzed.[Result]The phycobiliprotein was rich in amino acids,which not only contained 18 kinds of essential amino acids,but also contained some non-essential amino acids like glycine,aspartic acid,etc.;Analysis on signal peptides and trails-membrane topological structure showed that the phycobiliprotein belonged to intracellular protein;Analysis on hydrophobicity/hydrophilicity showed that the phycobiliprotein belonged to hydrophilic protein;Phylogenetic analysis showed that the phycobiliprotein had a high homology with Arthrospira,which reached 99%-100%.[Conclusion]The study provided a certain reference for studying the relationship and interaction between α subunit and β subunit.展开更多
基金supported by the National Natural Science Foundation of China(32100773 and U20A6005)the National Science and Technology Innovation 2030-Major Project of China(2021ZD0202500)+4 种基金Shenzhen Medical Research Fund(B2402024)China Postdoctoral Science Foundation(2021M693296)Shenzhen Science and Technology Program(JCYJ20230807093815032)Guangdong High-level Hospital Construction Fund(ynkt2021-zz33 and LCYJ2022093)the Natural Science Foundation of Guangdong Province,China(2022A1515010297).
文摘Dear Editor,The Cay2.1 channel,also known as the P/Q-type Ca^(2+) channel,is a particular type of voltage-gated Ca^(2+) channel primarily expressed on the presynaptic membrane in the brain[1].It serves as an essential part of the precisely orchestrated neurotransmitter release machinery.
基金supported by a grant from Ministry of Science and Technology China,No.2022ZD0204704(to WW)the National Natural Science Foundation of China,No.82301572(to XZ)the China Postdoctoral Science Foundation,No.2023M731202(to XZ)。
文摘Nonhuman primates are increasingly being used as animal models in neuroscience research.However,efficient neuronal tracing techniques for labeling motor neurons and primary sensory afferents in the monkey spinal cord are lacking.Here,by injecting the cholera toxin B subunit into the sciatic nerve of a rhesus monkey,we successfully labeled the motor neurons and primary sensory afferents in the lumbar and sacralspinal cord.Labeled alpha motor neurons were located in lamina IX of the L6–S1 segments,which innervate both flexors and extensors.The labeled primary sensory afferents were mainly myelinated Aβfibers that terminated mostly in laminae I and II of the L4–L7 segments.Together with the labeled proprioceptive afferents,the primary sensory afferents formed excitatory synapses with multiple types of spinal neurons.In summary,our methods successfully traced neuronal connections in the monkey spinal cord and can be used in spinal cord studies when nonhuman primates are used.
基金supported by the National Natural Science Foundation of China(No.81974553)the High Level Key Disciplines of Traditional Chinese Medicine:Basic Theory of Traditional Chinese Medicine,National Administration of Traditional Chinese Medicine(No.zyyzdxk-2023118)+3 种基金the Special Funding for the Taishan Scholars Project(Nos.tsqn202211137,tsqn201909186,and tsqn202211355)the Natural Science Foundation of Shandong Province(Nos.ZR2020ZD17 and ZR2021LZY018)the City-School Integration Development Strategy Project of Jinan City(No.JNSX2023056)the Chinese Medicine and Brain Science Youth Scientific Research Innovation Team,Shandong University of Traditional Chinese Medicine(No.22202101),China.
文摘Premenstrual dysphoric disorder(PMDD),a subtype of premenstrual syndrome(PMS),involves physical and emotional symptoms that impact patients'daily lives and productivity.A reliable,side-effect-free clinical intervention is needed.Shuyu capsule is an effective traditional Chinese medicine preparation for PMDD used in the clinics,but its therapeutic mechanism remains unclear.Previous research has suggested that theγ-aminobutyric acidergic(GABAergic)system in the periaqueductal gray(PAG)may play a role in treating PMDD with traditional Chinese medicine,but there is a lack of functional verification.This study aims to reveal the potential mechanism of the Shuyu capsule in treating PMDD.The study employed an experimental design using female C57BL/6J and Vgat-Cre mice to assess the effects of Shuyu capsules on PMDD,with a focus on the GABAergic system in the dorsal PAG(dPAG).Assessments were conducted using the forced swimming test(FST)to gauge depression-like behaviors and western blot(WB)and immunofluorescence(IF)to measure the numbers of active GABAergic neurons and theγ-aminobutyric acid type A receptor(GABA,R)δsubunit(GABRD)expression.Chemogenetic techniques and adeno-associated virus were specifically used to activate GABAergic neurons and knock down the expression of subunits,respectively,providing insights into the neurobiological mechanisms underpinning the therapeutic effects of Shuyu capsules in treating PMDD.After being stressed by FST,the immobility duration of PMDD mice in the late dioestrus(LD)phase decreased after the Shuyu capsule intervention,implying that it can improve the estrous cycle-dependent depression-like phenotype in PMDD mice.Additionally,the application of Shuyu capsule can downregulate the expression of GABRD and reverse the downtrend of activated GABAergic neurons in the dPAG of PMDD model mice.We also found that single-target manipulation was enough to improve the depressionlike behavior of PMDD model mice.Transgenic mice with GABRD knockout were established,and their behaviors were tested,revealing changes in their exploratory behaviors,indicating that the GABRD may be closely related to anxiety disorders.Shuyu capsule plays an anti-PMDD role by activating GABAergic neurons and downregulating the expression of GABRD in the dPAG.This provides a theoretical basis for the clinical treatment of PMDD with traditional Chinese medicine and promotes the development of drugs for treating PMDD.
基金Supported by National Natural Science Foundation of China,No.82160762Guangxi Zhuang Autonomous Region Administration of Traditional Chinese Medicine Scientific Research Project,No.GXZYA20230267+2 种基金China Undergraduate Innovation and Entrepreneurship Training Program,No.S202410598060XChina Undergraduate Innovation and Entrepreneurship Training Program,No.X202410598360Future Academic Star of Guangxi Medical University,No.WLXSZX24074.
文摘BACKGROUND In recent years,many studies have shown that proteasome 26S subunit non-ATPase 6(PSMD6)plays an important role in the occurrence and development of malignant tumours.Unfortunately,there are no reports on the evaluation of the potential role of PSMD6 in hepatocellular carcinoma(HCC).AIM To comprehensively evaluate the overexpression pattern and clinical significance of PSMD6 in HCC tissues.METHODS This study integrated PSMD6 mRNA expression profiles from 4672 HCC and 3667 non-HCC tissues,along with immunohistochemical scores from 383 HCC and adjacent tissues,to assess PSMD6 overexpression in HCC.Clustered regularly interspaced short palindromic repeats knockout technology evaluated PSMD6’s essential role in HCC cell growth.Functional enrichment analysis explored the molecular mechanism of PSMD6 abnormalities in HCC.Drug sensitivity analysis and molecular docking analysed the effect of abnormal expression of PSMD6 on the drug sensitivity of HCC cells.RESULTS The results of 41 external and two internal datasets showed that PSMD6 mRNA(SMD=0.26,95%CI:0.09-0.42,P<0.05)and protein(SMD=2.85,95%CI:1.19-4.50,P<0.05)were significantly overexpressed in HCC tissues.The integrated analysis results showed that PSMD6 had a significant overexpression pattern in HCC tissues(SMD=0.40,95%CI:0.15-0.66,P<0.05).PSMD6 knockout inhibited HCC cell growth(chronos scores<-1).Functional enrichment implicated ribosome biogenesis and RNA splicing.Significant enrichment of signalling pathways such as RNA degradation,ribosomes,and chemical carcinogenesis—reactive oxygen species.Drug sensitivity analysis and a molecular docking model showed that high expression of PSMD6 was associated with the tolerance of HCC cells to drugs such as ML323,sepantronium bromide,and GDC0810.Overexpressed PSMD6 effectively distinguished HCC tissues(AUC=0.75,95%CI:0.71-0.79).CONCLUSION This study was the first to discover that PSMD6 was overexpressed in HCC tissues.PSMD6 is essential for the growth of HCC cells and may be involved in ribosome biogenesis and RNA splicing.
基金supported by the Biological Breeding-National Science and Technology Major Project(2023ZD0406804,2023ZD0402701)Major Project of Hubei Hongshan Laboratory(2022hszd019)First-Class Discipline Construction Funds of the College of Plant Science and Technology at Huazhong Agricultural University(2022ZK PY002)。
文摘Improving protein quality and grain yield traits coordinately is an important goal for crop breeding.To date,many protein-quality or grain-yield regulation genes have been identified.However,the genetic strategies integrating these genes in good-protein-quality and high-yield crop breeding practice are far from established.Here,we characterized the functions of the MADS domain-containing protein Zm MADS8 and Zea mays G protein gamma subunit 1(Zm GG1)in regulating protein quality and grain yield of maize.Zm MADS8 positively regulates zein protein accumulation and negatively regulates nonzein protein and lysine levels in kernels by interacting with Zm MADS47 to promote the transcriptional activation of Opaque2.Additionally,Zm MADS8 regulates starch content of kernels by targeting genes involved in starch biosynthesis.Zm GG1,a putative interactor of Zm MADS8,negatively regulates kernel number with a trade-off effect on kernel starch accumulation.The mads8;zmgg1 double mutant improved protein quality by attenuating zein biosynthesis and increasing essential lysine level,and increased grain yield by increasing kernel number,compensating for decreased starch biosynthesis.Our findings revealed the biological function of Zm MADS8 and Zm GG1 in regulating protein quality and yield related traits and suggested a genetic strategy by direct editing of Zm MADS8 and Zm GG1 to improve grain yield and protein quality simultaneously.
基金Supported by Shandong Provincial Natural Science Foundation,No.ZR2023MH329Project of Shandong Province Higher Educational Youth Innovation Science and Technology Program,No.2023KJ263and Natural Science Foundation of Gansu Province,China,No.22JR5RA953.
文摘BACKGROUND As a member of the chaperonin-containing tailless complex polypeptide 1(TCP1)complex,which plays a pivotal role in ensuring the accurate folding of numerous proteins,chaperonin-containing TCP1 subunit 6A(CCT6A)participates in various physiological and pathological processes.However,its effects on cell death and cancer therapy and the underlying mechanisms need further exploration in colorectal cancer(CRC)cells.AIM To explore the effects of CCT6A on cell death and cancer therapy and the underlying mechanisms in CRC.METHODS Cell proliferation was evaluated using the MTS assay,EdU staining,and colony growth assays.The expression of CCT6A was monitored by immunoblotting and quantitative PCR.CCT6A was knocked out by CRISPR-Cas9,and overexpressed by transfecting plasmids.Autophagy was examined by immunoblotting and the mCherry-GFP-LC3 assay.To monitor apoptosis and necroptosis,immunoblotting,co-immunoprecipitation,and flow cytometry were employed.RESULTS Cisplatin(DDP)exerted cytotoxic effects on CRC cells while simultaneously downregulating the expression of CCT6A.Depletion of CCT6A amplified the cytotoxic effects of DDP,whereas overexpression of CCT6A attenuated these adverse effects.CCT6A suppressed autophagy,apoptosis,and necroptosis under both basal and DDP-treated conditions.Autophagy inhibitors significantly enhanced the cytotoxic effects of DDP,whereas a necroptosis inhibitor partially reversed the cell viability loss induced by DDP.Furthermore,inhibiting autophagy enhanced both apoptosis and necroptosis induced by DDP.CONCLUSION CCT6A negatively modulates autophagy,apoptosis,and necroptosis,and CCT6A confers resistance to DDP therapy in CRC,suggesting its potential as a therapeutic target.
文摘BACKGROUND Hepatocellular carcinoma(HCC)is a leading cause of cancer-related mortality worldwide,with hepatitis B virus(HBV)infection serving as a significant etiological factor in endemic regions.Alpha-fetoprotein(AFP),the most commonly used biomarker,has limited sensitivity,particularly in AFP-negative HCC.Recent studies have identified origin recognition complex subunit 1(ORC1)and extra spindle pole bodies-like 1(ESPL1)as promising serum biomarkers,both linked to HBV DNA integration,a mechanism known to drive hepatocarcinogenesis.AIM To assess serum ORC1’s diagnostic value for HBV-HCC and its link to S gene integration.METHODS In this case-control study,479 HBV-infected patients were enrolled,including 20 hepatitis B,154 with HBV-related cirrhosis,and 96 with HBV-HCC.The control group comprised 73 individuals:29 with non-HBV-HCC and 44 healthy participants.Serum ORC1 and ESPL1 were measured by enzyme-linked immunosorbent assay.HBV integration sites were identified via whole-genome sequencing.Diagnostic performance was assessed using receiver operating characteristic analysis,including in AFP-negative patients.RESULTS HBV integration near the ORC1 locus(chromosome 1p32.3)was detected in 71.4%of HBV-HCC tissues.Serum ORC1 levels were significantly higher in HBV-infected patients than in non-HBV-infected controls(980.11 ng/L vs 746.82 ng/L,P<0.05)and in HBV-HCC compared with non-HBV-HCC(1077.07 ng/L vs 749.54 ng/L,P<0.05).Serum ORC1 and ESPL1 were elevated in HBV-HCC regardless of AFP status,and detected 64.8%and 73.2%of AFP-negative cases,respectively.The combined panel of ORC1[Area under receiver operating characteristic curve(AUC)=0.587],ESPL1(AUC=0.776),and AFP(AUC=0.844)achieved an AUC of 0.887,significantly higher than any single marker(P<0.05),with a sensitivity of 84.44%,specificity of 84.19%,and a negative predictive value of 94.91%.CONCLUSION Serum ORC1,driven by HBV integration,is a promising biomarker especially for AFP-negative HBV-HCC.Its combination with ESPL1 and AFP significantly improves early detection.
基金Supported by the National Natural Science Foundation of China,No.82160213 and No.U22A2022the Guangxi Natural Science Foundation,No.2023GXNSFAA026029,No.2024GXNSFBA010059,and No.2024GXNSFAA010079。
文摘BACKGROUND Laryngeal squamous cell carcinoma(LSCC)is a prevalent head and neck malignancy with suboptimal survival rates due to late detection and therapeutic resistance.AIM To investigate chaperonin-containing TCP1 subunit 3(CCT3)expression and its clinical implications,and its effects on LSCC cell growth.METHODS Systematic data on CCT3 mRNA expression were collected from biomedical databases,and integrated further based on the standardized mean difference and the summary receiver operating characteristic curve.Single-cell RNA-seq data were mined to validate the expression level of CCT3 mRNA.In-house immunohistochemistry was performed to explore the CCT3 protein levels of clinical LSCC samples and their relationship with clinical parameters.The growth function of LSCC cell was analyzed using CRISPR knockout screening.CCT3-related signaling pathway analyses were conducted using gene set enrichment analysis.Protein-protein interaction network construction was performed to identify hub genes.RESULTS CCT3 mRNA was significantly overexpressed in 269 LSCC tissues cases across multiple independent datasets(standardized mean difference=32,area under the curve=0.93);At the translational level,the in-house immunohistochemical analysis further demonstrated the consistent upregulation of CCT3 protein in 88 cases of LSCC samples(58 non-LSCC samples vs 30 LSCC samples,P=1.4e^(-14)).Analysis of clinical parameters showed no significant differences among subgroup.Functional characterization with clustered regularly interspaced short palindromic repeats--mediated gene knockout revealed that depletion of CCT3 potently suppressed LSCC cell viability in vitro.Gene set enrichment analysis indicated that CCT3 was markedly associated with several key oncogenic pathways,including extracellular matrix receptor interaction and cell cycle regulation pathways.CONCLUSION CCT3 upregulation in LSCC may influence cellular growth by regulating related pathways,indicating its potential as a biomarker and therapeutic target for LSCC.
文摘A deficiency ofγδT cells has been described in Crohn's disease(CD).AIM To analyze the gene expression of interleukin 7(IL-7)and its receptors in the tissues of patients with CD.METHODS We studied the peripheral blood of 80 patients with CD,comparing them with a group of 80 healthy subjects.The number and apoptosis ofαβandγδT cells in peripheral blood and the proportion ofαβandγδT cells in the intestinal tissues of patients with CD(n=25)were studied.The gene and protein expression of IL-7,IL-2 receptor subunitγ[cluster of differentiation 132(CD132)],receptorα(CD127),and caspase-3 in tissues was analyzed by quantitative PCR.Serum IL-7 levels were also analyzed.RESULTS In patients with CD,a decreased number ofγδT cells and an increase in the apoptosis of CD56+αβandγδT cells in peripheral blood was observed(P<0.0001 and P<0.01)respectively,and there was an inverse correlation among T subsets and their apoptosis.In addition,IL-7 gene expression and IL-7 protein in the tissues of these patients were increased.The titers of caspase-3 in tissues were low vs control group(P>0.01).The percentage of CD8+γδT cells decreased in tissues(P<0.01),and was directly related to IL-7 levels in peripheral blood.The expression of IL-2 receptor subunitγ(CD132)was greatly decreased in the tissues of patients with CD(P<0.05).CONCLUSION There may be a cause-effect relationship between the lower gene expression of the IL-2 receptor subunitγ(CD132)in tissues of patients with CD andγδT cells immunodeficiency.
基金supported by the National Natural Science Foundation of China(No.82471389,No.32470986,No.82271385)Natural Science Foundation of Guangdong Province(No.2024A1515010471).
文摘Mosquito-borne flaviviruses,such as Zika virus(ZIKV)and dengue virus(DENV),cause diverse severe clinical manifestations including fever,rash,hepatitis,arthralgia,and congenital anomalies.Here,we identified a host factor,the adaptor protein complex 1 gamma 1 subunit(AP1G1),which plays an important role in both ZIKV and dengue virus 2(DENV2)infections.We explored the role of AP1G1 in ZIKV and DENV2 infections using CRISPR/Cas9 gene editing technology and RNA interference(RNAi)techniques.Knockout or silencing of AP1G1 decreases the replication of ZIKV and DENV2 in multiple human cell lines.Intriguingly,depletion of AP1G1 results in a significant reduction in ZIKV at an early stage,but decreases DENV2 replication levels during the late stage,suggesting that AP1G1 plays distinct roles in the infection by ZIKV and DENV2.Furthermore,we determined that AP1G1 mediates ZIKV-endosomal membrane fusion through inhibitor experiments and fluorescence labeling assays.Mechanistically,we found that AP1G1 exerts its pro-viral effect through binding to the ZIKV envelope glycoprotein(E protein).This interaction promotes the fusion of viral and endosomal membranes,during which the ZIKV genomic RNAs are released from the endosome into the cytoplasm,a process that facilitates viral replication.However,for DENV2 infection,AP1G1 primarily affects its viral RNA replication stage,rather than the fusion of virus-endosomal membrane.Taken together,our work demonstrates that AP1G1 plays a pro-viral role in both ZIKV and DENV2 infections via distinct mechanisms,highlighting its potential as a therapeutic target for antiviral strategies.
文摘Crohn’s disease(CD)is a chronic inflammatory disorder characterized by dysregulated immune responses and significant disruption of intestinal immunity.A recent case-control study by Andreu-Ballester et al revealed decreased expression of interleukin(IL)-2 receptor subunitγ(CD132)in CD tissues,a finding that has profound implications for understanding immune dysregulation in CD.CD132,an essential component of the IL-7/IL-2 signaling axis,is critical forγδT cell survival and function,which are pivotal for maintaining gut integrity and modulating inflammation.Here,we propose that reduced CD132 expression represents a key mechanism underlyingγδT cell deficiencies in CD,contributing to impaired immune surveillance and exacerbated inflammation.This hypothesis integrates emerging evidence from cytokine signaling and immunopathology in CD,offering new insights into its pathogenesis.These findings highlight the therapeutic potential of targeting the IL-7/IL-2 axis to restore immune homeostasis in CD,presenting a novel avenue for future research and intervention.
文摘Nicotinic acetylcholine receptors (nAChRs) play a significant role in excitatory synaptic transmission in insects and are the target for chloronicotinyl and nereistoxin insecticides.In recent years,Chilo suppressalis,an economically important pest of rice,developed high resistance against monosultap,a nereistoxin insecticide acting on nAChR.In order to reveal the hypothesized target insensitive mechanism,studies on the molecular property of nAChR from Chilo suppressalis are required.In this study,the full length cDNA of nAChR α subunit from this pest was cloned by RT-PCR.Sequence analysis shows that it is a novel nAChR α subunit,which was named as Cs α 1(Genbank accession No.AF418987).It contains 1?997?bp nucleotides and involves an open reading frame (ORF) encoding a mature protein of 509 amino acids excluding a signal peptide of 24 amino acids.The deduced amino acid sequence was 52%-94% identical to the reported insect nAChR genes.
文摘A vacuolar ATPase (V-ATPase.) B subunit gene has been cloned and characterized front a phosphorus starvation induced rice root subtractive cDNA library by suppression subtractive hybridization (SSH) method and RT-PCR amplification. This gene encodes a polypeptide of 487 amino acid residues, containing a conservative ATP binding site and with a molecular weight of 54.06 kD and an isoelectric point of 4.99, southern analysis of the. genomic DNA indicates that V-ATPase B subunit is encoded by a single gene in rice genome. The amino acid homologies of V-ATPase B subunits among different organisms range from 76% to 97% and reveals that the evolution of V-ATPase B subunit is accompanied with the biological evolution. Expression pattern analysis indicated that the maximal expression of V-ATPase B subunit gene occurred at an early stage (6 - 12 h) after phosphorus starvation in roots, and lately stage (24 - 48 It) in leaves. Under phosphorus deficiency, the up-regulated expression of V-ATPase gene was presumed to strengthen the proton transport and provide the required energy to maintain an electrochemical gradient across the tonoplast to facilitate Phosphorus transport.
基金Supported by the Special Project for Public-welfare Industry of Chinese Forestry (200704025) The Key Project of Chinese Ministry of Education (102023)~~
文摘[Objective]The α subunit gene of phycobiliprotein from Spirulina maxima was studied in order to provide a basis for the subsequent study of phycobiliprotein.[Method] Amino acids composition,signal peptides,hydrophobicity/hydrophilicity and trails-membrane topological structure of α subunit gene of phycobiliprotein from Spirulina maxima which registered in GenBank（GenBank AF441177） were analyzed and predicted by the tools of bioinformatic analysis.Meanwhile,phylogenetic tree was constructed based on α subunit gene of phycobiliprotein from Spirulina maxima,and its molecular evolution was also analyzed.[Result]The phycobiliprotein was rich in amino acids,which not only contained 18 kinds of essential amino acids,but also contained some non-essential amino acids like glycine,aspartic acid,etc.;Analysis on signal peptides and trails-membrane topological structure showed that the phycobiliprotein belonged to intracellular protein;Analysis on hydrophobicity/hydrophilicity showed that the phycobiliprotein belonged to hydrophilic protein;Phylogenetic analysis showed that the phycobiliprotein had a high homology with Arthrospira,which reached 99%-100%.[Conclusion]The study provided a certain reference for studying the relationship and interaction between α subunit and β subunit.
