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
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
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
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
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
BACKGROUND Ulcerative colitis(UC)is a chronic,nonspecific intestinal inflammatory disease with undefined pathogenesis.Non-SMC condensin I complex subunit D2(NCAPD2)and non-SMC condensin II complex subunit D3(NCAPD3)pl...BACKGROUND Ulcerative colitis(UC)is a chronic,nonspecific intestinal inflammatory disease with undefined pathogenesis.Non-SMC condensin I complex subunit D2(NCAPD2)and non-SMC condensin II complex subunit D3(NCAPD3)play pivotal roles in chromosome assembly and segregation during both mitosis and meiosis.To date,there has been no relevant report about the functional role of NCAPD2 and NCAPD3 in UC.AIM To determine the level of NCAPD2/3 in intestinal mucosa and explore the mechanisms of NCAPD2/3 in UC.METHODS Levels of NCAPD2/3 in intestinal tissue were detected in 30 UC patients and 30 healthy individuals with in situ hybridization(ISH).In vitro,NCM60 cells were divided into the NC group,model group,si-NCAPD2 group,si-NCAPD3 group and si-NCAPD2+si-NCAPD3 group.Inflammatory cytokines were measured by ELISA,IKK and NF-κB were evaluated by western blot,and IKK nucleation and NF-κB volume were analyzed by immunofluorescence assay.RESULTS Compared with expression in healthy individuals,NCAPD2 and NCAPD3 expression in intestinal tissue was significantly upregulated(P<0.001)in UC patients.Compared with levels in the model group,IL-1β,IL-6 and TNF-αin the si-NCAPD2,si-NCAPD3 and si-NCAPD2+si-NCAPD3 groups were significantly downregulated(P<0.01).IKK and NF-κB protein expression in the si-NCAPD2,si-NCAPD3 and si-NCAPD2+si-NCAPD3 groups was significantly decreased(P<0.01).Moreover,IKK nucleation and NF-κB volume were suppressed upon si-NCAPD2,si-NCAPD3 and si-NCAPD2+si-NCAPD3 transfection.CONCLUSION NCAPD2/3 is highly expressed in the intestinal mucosa of patients with active UC.Overexpression of NCAPD2/3 promotes the release of pro-inflammatory cytokines by modulating the IKK/NF-κB signaling pathway.展开更多
[Objective] The research aimed to study the effects of microwave on the chick embryo development and the cognitive function of chickling. [Method] The microwave which was transmitted by the permatron and was 2 450 MHz...[Objective] The research aimed to study the effects of microwave on the chick embryo development and the cognitive function of chickling. [Method] The microwave which was transmitted by the permatron and was 2 450 MHz was used to simulate the microwave radiation source to radiate the hatching eggs until the chickling was hatched out. The disposable passive avoidance learning and RT-PCR were respectively used to detect the influences of microwave on the cognitive function of chickling and the expression amounts of NMDA receptor NR1 and NR2 subunits. [Result] After the microwave radiation,the avoidance rate of exposed group was significantly lower than that in the control group. Especially the avoidance rate of highest radiation intensity group was extremely significantly lower than that in the control group. Meanwhile,the body weights of two groups of chickling in the exposed group increased,and the hatching time in one group increased. Via RT-PCR analysis,the expression amount of NR2 subunit increased on the 10th day and the 15th day. The expression amount of NR1 subunit only decreased on the 15th day. [Conclusion] The microwave had the certain influence on the individual development. By changing the structure composition and function of NMDA receptor in the endbrain,the microwave made the self-regulation ability of chickling decline,which had the certain damage on the cognitive function.展开更多
Serine/threonine phosphatase calcineurin(CN)is a unique but confounding calcium/calmodulin-mediated enzyme,which is composed of a catalytic subunit A(CNA)and a regulatory subunit B(CNB).We cloned six transcripts for C...Serine/threonine phosphatase calcineurin(CN)is a unique but confounding calcium/calmodulin-mediated enzyme,which is composed of a catalytic subunit A(CNA)and a regulatory subunit B(CNB).We cloned six transcripts for CNA named from NlCNA-X1 to NlCNA-X6,one CNB named NlCNB1 and one CNB homologous gene NlCNBH1 from Nilaparvata lugens.All of them are constitutively transcripted in various tissues and developmental stages.The primary structure of the six isoforms showed obvious differences in the length and composition of the amino acid sequence between the two binding domains of Ca^(2+)/calmodulin(CaM)and CNB.Ca^(2+)-binding EF-hand motifs were found in NlCNB1 and NlCNBH1.The specific gene silencing of NlCNA,NlCNB1 and NlCNBH1 respectively by RNAi resulted in drastical reduction in survival rate,female weight,eclosion rate and fecundity of N.lugens.These results showed that NlCNA,NlCNB1 and NlCNBH1 were required for N.lugens growth and reproduction.The negative effects of NlCNB1 silence on nymph mortality(97%),molting malformation(90%)and female sterile(50%)were more serious than those of NlCNA or NlCNBH1.qRT-PCR and enzyme-linked immunosorbent assay(ELISA)analyses indicated that the nymphs with silenced NlCNA,NlCNB1 or NlCNBH1 showed impaired hormone and energy metabolism.In nymphs,the contents of 20-hydroxyecdysone(20E)after NlCNB1 RNAi and phenoloxidase after NlCNA RNAi were particularly decreased.These results suggested that NlCNA is involved in immunity of N.lugens by regulation of phenoloxidase,while NlCNB1 may control the growth and development of N.lugens by 20E signaling pathway in addition to interact with CNA.Injection of 70 ng/μL dsNlCNB1 resulted in 77.0%down regulation of NlCNB1,and the nymph mortality was up to 57.9%at 10 d after injection.Therefore,NlCNB1 could be a potential candidate target used for strategy design in control of N.lugens.Our results revealed the importance of CN in the regulation of the growth and development of N.lugens,which provided a basis for further study of the molecular mechanism of CN.展开更多
[Objective] This study aimed to clone and analyze the cDNA of ATPase β subunit gene from Eleutherococcus senticosus.[Method] A pair of homologous primers was designed according to the chloroplast ATPase β subunit ge...[Objective] This study aimed to clone and analyze the cDNA of ATPase β subunit gene from Eleutherococcus senticosus.[Method] A pair of homologous primers was designed according to the chloroplast ATPase β subunit gene sequences of the known species;then the gene cDNA of E.senticosus were amplified by RT-PCR and compared with that of the known species;its structure was predicted finally.[Result] 1 099 bp of ATPase beta subunit cDNA of E.senticosus which encodes 366 amino acids was amplified by RT-PCR.Sequence comparison and structure prediction showed that amino acids encoded by the ATPase beta subunit gene of E.senticosus shared the highest homology,up to 96.41% with that of Oryza sativa.In the secondary structure,the protein contained 171 alpha helixes accounting for 46.72%,53 extended strands accounting for 14.48%,27 beta sheets accounting for 7.38% and 115 random coils which took up 31.42%.The amino acids 262-271 were the symbolic site of ATPase β subunit.The whole peptide chain had no obvious hydrophobic region and was primarily confirmed as a hydrophilic protein.[Conclusion] The cNDA of ATPase β subunit gene cloned from E.senticosus in this study will provide reference for learning the effect of energy metabolism on secondary metabolism,structure and function of ATPase in E.senticosus.展开更多
In order to evaluate the immune effect of the protein expressed by the universal vector pET-mLTA-CTLA-4 plus IBDV subunit, the fusion protein mLTA-CTLA-4 was expressed and purified. Protein toxicity tests were carried...In order to evaluate the immune effect of the protein expressed by the universal vector pET-mLTA-CTLA-4 plus IBDV subunit, the fusion protein mLTA-CTLA-4 was expressed and purified. Protein toxicity tests were carried out on rabbits.The VP2 gene of infectious bursal virus was amplified by RT-PCR, and lately used for pET-VP2 construction. Ten-day-old free healthy chickens were chosen for a grouped test, including the mLTA-CTLA-4（at different doses） plus VP2 groups, IBDV living vaccine group and control group. Serum and mucosal samples were collected regularly and the neutralization titers of IgG and IgA were assayed, while an animal protection test was conducted to determine the protection rate. The results showed that the protein m LTA-CTLA-4 was non-toxic and its protection rate was100%. IgG or IgA levels in the IBDV vaccine group were slightly higher than those in recombinant protein groups. These results indicated that the recombinant protein mLTA-CTLA-4 could be applied with IBDV subunit vaccine to protect chickens from infection.展开更多
The high-molecular-weight (HMW) glutenin subunits and their coding genes from Aegilops umbellulata Zhuk. (UU, 2n = 2x = 14) were characterized using SDS-PAGE analysis and molecular approaches. SDS-PAGE analysis showed...The high-molecular-weight (HMW) glutenin subunits and their coding genes from Aegilops umbellulata Zhuk. (UU, 2n = 2x = 14) were characterized using SDS-PAGE analysis and molecular approaches. SDS-PAGE analysis showed that the 1Ux subunits from four different accessions possessed electrophoretic mobilities close to, or slower than, that displayed by the 1Dx2.2 subunit of common wheat. The electrophoretic mobilities of the 1Uy subunits were generally similar to those shown by the 1Dy subunits of common wheat. The complete open reading frames of the 1Ux and 1Uy genes were amplified by PCR and subsequently cloned and sequenced. Amino acid sequence comparisons suggested that the primary structure of the 1Ux and 1Uy subunits were identical to that of published HMW glutenin subunits from related species, Phylogenetic analysis indicated that the HMW glutenin subunits of Ae. umbellulata were most closely related to those encoded by the D genome of Triticeae.展开更多
Epilepsy is one of the most common neurological diseases. Of all cases, 70%–80% are considered to be due to genetic factors. In recent years, a large number of genes have been identified as being involved in epilepsy...Epilepsy is one of the most common neurological diseases. Of all cases, 70%–80% are considered to be due to genetic factors. In recent years, a large number of genes have been identified as being involved in epilepsy.Among them, N-methyl-D-aspartate receptor（NMDAR）subunit-encoding genes represent a large proportion, suggesting an important role for NMDARs in epilepsy. In this review, we summarize and analyze the genotypes, functional alterations, and clinical aspects of NMDAR subunit mutations/variants identified from patients with epilepsy.These data will help to throw light upon the pathogenicity of these NMDAR mutations and advance our understanding of the subtle and complicated role of NMDARs in epilepsy. It will also offer new insights into precision therapy for this disorder.展开更多
Autophagy has been suggested to participate in the pathology of hypoxic-ischemic brain damage（HIBD）.However,its regulatory role in HIBD remains unclear and was thus examined here using a rat model.To induce HIBD,the...Autophagy has been suggested to participate in the pathology of hypoxic-ischemic brain damage（HIBD）.However,its regulatory role in HIBD remains unclear and was thus examined here using a rat model.To induce HIBD,the left common carotid artery was ligated in neonatal rats,and the rats were subjected to hypoxia for 2 hours.Some of these rats were intraperitoneally pretreated with the autophagy inhibitor 3-methyladenine（10 m M in 10 μL） or the autophagy stimulator rapamycin（1 g/kg） 1 hour before artery ligation.Our findings demonstrated that hypoxia-ischemia-induced hippocampal injury in neonatal rats was accompanied by increased expression levels of the autophagy-related proteins light chain 3 and Beclin-1 as well as of the AMPA receptor subunit GluR 1,but by reduced expression of GluR 2.Pretreatment with the autophagy inhibitor 3-methyladenine blocked hypoxia-ischemia-induced hippocampal injury,whereas pretreatment with the autophagy stimulator rapamycin significantly augmented hippocampal injury.Additionally,3-methyladenine pretreatment blocked the hypoxia-ischemia-induced upregulation of Glu R1 and downregulation of GluR2 in the hippocampus.By contrast,rapamycin further elevated hippocampal Glu R1 levels and exacerbated decreased GluR2 expression levels in neonates with HIBD.Our results indicate that autophagy inhibition favors the prevention of HIBD in neonatal rats,at least in part,through normalizing Glu R1 and GluR2 expression.展开更多
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 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.
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.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.
基金Supported by National Natural Science Foundation of China,No.81673973Natural Science Foundation of Jiangsu Province,China,No.BK20161577the Developing Program for Highlevel Academic Talent from Jiangsu Hospital of Chinese Medicine,No.y2018rc16
文摘BACKGROUND Ulcerative colitis(UC)is a chronic,nonspecific intestinal inflammatory disease with undefined pathogenesis.Non-SMC condensin I complex subunit D2(NCAPD2)and non-SMC condensin II complex subunit D3(NCAPD3)play pivotal roles in chromosome assembly and segregation during both mitosis and meiosis.To date,there has been no relevant report about the functional role of NCAPD2 and NCAPD3 in UC.AIM To determine the level of NCAPD2/3 in intestinal mucosa and explore the mechanisms of NCAPD2/3 in UC.METHODS Levels of NCAPD2/3 in intestinal tissue were detected in 30 UC patients and 30 healthy individuals with in situ hybridization(ISH).In vitro,NCM60 cells were divided into the NC group,model group,si-NCAPD2 group,si-NCAPD3 group and si-NCAPD2+si-NCAPD3 group.Inflammatory cytokines were measured by ELISA,IKK and NF-κB were evaluated by western blot,and IKK nucleation and NF-κB volume were analyzed by immunofluorescence assay.RESULTS Compared with expression in healthy individuals,NCAPD2 and NCAPD3 expression in intestinal tissue was significantly upregulated(P<0.001)in UC patients.Compared with levels in the model group,IL-1β,IL-6 and TNF-αin the si-NCAPD2,si-NCAPD3 and si-NCAPD2+si-NCAPD3 groups were significantly downregulated(P<0.01).IKK and NF-κB protein expression in the si-NCAPD2,si-NCAPD3 and si-NCAPD2+si-NCAPD3 groups was significantly decreased(P<0.01).Moreover,IKK nucleation and NF-κB volume were suppressed upon si-NCAPD2,si-NCAPD3 and si-NCAPD2+si-NCAPD3 transfection.CONCLUSION NCAPD2/3 is highly expressed in the intestinal mucosa of patients with active UC.Overexpression of NCAPD2/3 promotes the release of pro-inflammatory cytokines by modulating the IKK/NF-κB signaling pathway.
基金Supported by Suzhou City Science and Technology Bureau Item(YJS0904)~~
文摘[Objective] The research aimed to study the effects of microwave on the chick embryo development and the cognitive function of chickling. [Method] The microwave which was transmitted by the permatron and was 2 450 MHz was used to simulate the microwave radiation source to radiate the hatching eggs until the chickling was hatched out. The disposable passive avoidance learning and RT-PCR were respectively used to detect the influences of microwave on the cognitive function of chickling and the expression amounts of NMDA receptor NR1 and NR2 subunits. [Result] After the microwave radiation,the avoidance rate of exposed group was significantly lower than that in the control group. Especially the avoidance rate of highest radiation intensity group was extremely significantly lower than that in the control group. Meanwhile,the body weights of two groups of chickling in the exposed group increased,and the hatching time in one group increased. Via RT-PCR analysis,the expression amount of NR2 subunit increased on the 10th day and the 15th day. The expression amount of NR1 subunit only decreased on the 15th day. [Conclusion] The microwave had the certain influence on the individual development. By changing the structure composition and function of NMDA receptor in the endbrain,the microwave made the self-regulation ability of chickling decline,which had the certain damage on the cognitive function.
基金This study was supported by the China Agriculture Research System(Grant No.CARS-01-38)Rice Pest Management Research Group of the Agricultural Science and Technology Innovation Program of China Academy of Agricultural Science(Grant No.CAAS-ASTIP-2016-CNRRI)+1 种基金Central Public-Interest Scientific Institution Basal Research Fund of China(Grant No.CPSIBRF-CNRRI-202122)Open Project Program of State Key Laboratory of Rice Biology,China(Grant No.20210302).
文摘Serine/threonine phosphatase calcineurin(CN)is a unique but confounding calcium/calmodulin-mediated enzyme,which is composed of a catalytic subunit A(CNA)and a regulatory subunit B(CNB).We cloned six transcripts for CNA named from NlCNA-X1 to NlCNA-X6,one CNB named NlCNB1 and one CNB homologous gene NlCNBH1 from Nilaparvata lugens.All of them are constitutively transcripted in various tissues and developmental stages.The primary structure of the six isoforms showed obvious differences in the length and composition of the amino acid sequence between the two binding domains of Ca^(2+)/calmodulin(CaM)and CNB.Ca^(2+)-binding EF-hand motifs were found in NlCNB1 and NlCNBH1.The specific gene silencing of NlCNA,NlCNB1 and NlCNBH1 respectively by RNAi resulted in drastical reduction in survival rate,female weight,eclosion rate and fecundity of N.lugens.These results showed that NlCNA,NlCNB1 and NlCNBH1 were required for N.lugens growth and reproduction.The negative effects of NlCNB1 silence on nymph mortality(97%),molting malformation(90%)and female sterile(50%)were more serious than those of NlCNA or NlCNBH1.qRT-PCR and enzyme-linked immunosorbent assay(ELISA)analyses indicated that the nymphs with silenced NlCNA,NlCNB1 or NlCNBH1 showed impaired hormone and energy metabolism.In nymphs,the contents of 20-hydroxyecdysone(20E)after NlCNB1 RNAi and phenoloxidase after NlCNA RNAi were particularly decreased.These results suggested that NlCNA is involved in immunity of N.lugens by regulation of phenoloxidase,while NlCNB1 may control the growth and development of N.lugens by 20E signaling pathway in addition to interact with CNA.Injection of 70 ng/μL dsNlCNB1 resulted in 77.0%down regulation of NlCNB1,and the nymph mortality was up to 57.9%at 10 d after injection.Therefore,NlCNB1 could be a potential candidate target used for strategy design in control of N.lugens.Our results revealed the importance of CN in the regulation of the growth and development of N.lugens,which provided a basis for further study of the molecular mechanism of CN.
基金Supported by the National Natural Science Foundation of China(30701086)the Natural Science Foundation of Hebei Province(C2009001252)~~
文摘[Objective] This study aimed to clone and analyze the cDNA of ATPase β subunit gene from Eleutherococcus senticosus.[Method] A pair of homologous primers was designed according to the chloroplast ATPase β subunit gene sequences of the known species;then the gene cDNA of E.senticosus were amplified by RT-PCR and compared with that of the known species;its structure was predicted finally.[Result] 1 099 bp of ATPase beta subunit cDNA of E.senticosus which encodes 366 amino acids was amplified by RT-PCR.Sequence comparison and structure prediction showed that amino acids encoded by the ATPase beta subunit gene of E.senticosus shared the highest homology,up to 96.41% with that of Oryza sativa.In the secondary structure,the protein contained 171 alpha helixes accounting for 46.72%,53 extended strands accounting for 14.48%,27 beta sheets accounting for 7.38% and 115 random coils which took up 31.42%.The amino acids 262-271 were the symbolic site of ATPase β subunit.The whole peptide chain had no obvious hydrophobic region and was primarily confirmed as a hydrophilic protein.[Conclusion] The cNDA of ATPase β subunit gene cloned from E.senticosus in this study will provide reference for learning the effect of energy metabolism on secondary metabolism,structure and function of ATPase in E.senticosus.
基金Supported by Jiangsu Provincial Postdoctoral Science Foundation(1401077B)Open Project of Jiangsu Provincial Key Laboratory of Veterinary Bio-pharmaceutical High-tech Research(JSKLKF1403)+3 种基金the Fenghuang Talent Engineering Project of Jiangsu Agrianimal Husbandry Vocational CollegeKey Project of Jiangsu Agri-animal Husbandry Vocational College(NSFZD1405)the Horizontal Cooperation Project of Yangzhou Chaotiange Agri-animal Husbandry Science and Technology Co.,Ltd.(00010114012,NSFPT201510)the Special Fund for Jiangsu Huaneng Medical Investment Co.,Ltd.(NSFPT201512)~~
文摘In order to evaluate the immune effect of the protein expressed by the universal vector pET-mLTA-CTLA-4 plus IBDV subunit, the fusion protein mLTA-CTLA-4 was expressed and purified. Protein toxicity tests were carried out on rabbits.The VP2 gene of infectious bursal virus was amplified by RT-PCR, and lately used for pET-VP2 construction. Ten-day-old free healthy chickens were chosen for a grouped test, including the mLTA-CTLA-4（at different doses） plus VP2 groups, IBDV living vaccine group and control group. Serum and mucosal samples were collected regularly and the neutralization titers of IgG and IgA were assayed, while an animal protection test was conducted to determine the protection rate. The results showed that the protein m LTA-CTLA-4 was non-toxic and its protection rate was100%. IgG or IgA levels in the IBDV vaccine group were slightly higher than those in recombinant protein groups. These results indicated that the recombinant protein mLTA-CTLA-4 could be applied with IBDV subunit vaccine to protect chickens from infection.
文摘The high-molecular-weight (HMW) glutenin subunits and their coding genes from Aegilops umbellulata Zhuk. (UU, 2n = 2x = 14) were characterized using SDS-PAGE analysis and molecular approaches. SDS-PAGE analysis showed that the 1Ux subunits from four different accessions possessed electrophoretic mobilities close to, or slower than, that displayed by the 1Dx2.2 subunit of common wheat. The electrophoretic mobilities of the 1Uy subunits were generally similar to those shown by the 1Dy subunits of common wheat. The complete open reading frames of the 1Ux and 1Uy genes were amplified by PCR and subsequently cloned and sequenced. Amino acid sequence comparisons suggested that the primary structure of the 1Ux and 1Uy subunits were identical to that of published HMW glutenin subunits from related species, Phylogenetic analysis indicated that the HMW glutenin subunits of Ae. umbellulata were most closely related to those encoded by the D genome of Triticeae.
基金supported by the grant of the National Basic Research Program of China (2014CB910300)
文摘Epilepsy is one of the most common neurological diseases. Of all cases, 70%–80% are considered to be due to genetic factors. In recent years, a large number of genes have been identified as being involved in epilepsy.Among them, N-methyl-D-aspartate receptor（NMDAR）subunit-encoding genes represent a large proportion, suggesting an important role for NMDARs in epilepsy. In this review, we summarize and analyze the genotypes, functional alterations, and clinical aspects of NMDAR subunit mutations/variants identified from patients with epilepsy.These data will help to throw light upon the pathogenicity of these NMDAR mutations and advance our understanding of the subtle and complicated role of NMDARs in epilepsy. It will also offer new insights into precision therapy for this disorder.
基金supported by the National Natural Science Foundation of China,No.81471488,81271378,81502157,and 81501291the Key Medical Subjects of Jiangsu Province of China,No.XK201120+3 种基金the Jiangsu Province Key Research and Development of Special Funds in China,No.BE2015644the Science and Technology Project of Suzhou City of China,No.SYSD2013105,SYS201446,SYS201441the Public Health Technology Project of Suzhou City of China,No.SS201536the Department of Pediatrics Clinical Center of Suzhou City of China,No.Szzx201504
文摘Autophagy has been suggested to participate in the pathology of hypoxic-ischemic brain damage（HIBD）.However,its regulatory role in HIBD remains unclear and was thus examined here using a rat model.To induce HIBD,the left common carotid artery was ligated in neonatal rats,and the rats were subjected to hypoxia for 2 hours.Some of these rats were intraperitoneally pretreated with the autophagy inhibitor 3-methyladenine（10 m M in 10 μL） or the autophagy stimulator rapamycin（1 g/kg） 1 hour before artery ligation.Our findings demonstrated that hypoxia-ischemia-induced hippocampal injury in neonatal rats was accompanied by increased expression levels of the autophagy-related proteins light chain 3 and Beclin-1 as well as of the AMPA receptor subunit GluR 1,but by reduced expression of GluR 2.Pretreatment with the autophagy inhibitor 3-methyladenine blocked hypoxia-ischemia-induced hippocampal injury,whereas pretreatment with the autophagy stimulator rapamycin significantly augmented hippocampal injury.Additionally,3-methyladenine pretreatment blocked the hypoxia-ischemia-induced upregulation of Glu R1 and downregulation of GluR2 in the hippocampus.By contrast,rapamycin further elevated hippocampal Glu R1 levels and exacerbated decreased GluR2 expression levels in neonates with HIBD.Our results indicate that autophagy inhibition favors the prevention of HIBD in neonatal rats,at least in part,through normalizing Glu R1 and GluR2 expression.
