Border-associated macrophages are located at the interface between the brain and the periphery, including the perivascular spaces, choroid plexus, and meninges. Until recently, the functions of border-associated macro...Border-associated macrophages are located at the interface between the brain and the periphery, including the perivascular spaces, choroid plexus, and meninges. Until recently, the functions of border-associated macrophages have been poorly understood and largely overlooked. However, a recent study reported that border-associated macrophages participate in stroke-induced inflammation, although many details and the underlying mechanisms remain unclear. In this study, we performed a comprehensive single-cell analysis of mouse border-associated macrophages using sequencing data obtained from the Gene Expression Omnibus(GEO) database(GSE174574 and GSE225948). Differentially expressed genes were identified, and enrichment analysis was performed to identify the transcription profile of border-associated macrophages. CellChat analysis was conducted to determine the cell communication network of border-associated macrophages. Transcription factors were predicted using the ‘pySCENIC' tool. We found that, in response to hypoxia, borderassociated macrophages underwent dynamic transcriptional changes and participated in the regulation of inflammatory-related pathways. Notably, the tumor necrosis factor pathway was activated by border-associated macrophages following ischemic stroke. The pySCENIC analysis indicated that the activity of signal transducer and activator of transcription 3(Stat3) was obviously upregulated in stroke, suggesting that Stat3 inhibition may be a promising strategy for treating border-associated macrophages-induced neuroinflammation. Finally, we constructed an animal model to investigate the effects of border-associated macrophages depletion following a stroke. Treatment with liposomes containing clodronate significantly reduced infarct volume in the animals and improved neurological scores compared with untreated animals. Taken together, our results demonstrate comprehensive changes in border-associated macrophages following a stroke, providing a theoretical basis for targeting border-associated macrophages-induced neuroinflammation in stroke treatment.展开更多
BACKGROUND The complexity of the immune microenvironment has an impact on the treatment of colorectal cancer(CRC),one of the most prevalent malignancies worldwide.In this study,multi-omics and single-cell sequencing t...BACKGROUND The complexity of the immune microenvironment has an impact on the treatment of colorectal cancer(CRC),one of the most prevalent malignancies worldwide.In this study,multi-omics and single-cell sequencing techniques were used to investigate the mechanism of action of circulating and infiltrating B cells in CRC.By revealing the heterogeneity and functional differences of B cells in cancer immunity,we aim to deepen our understanding of immune regulation and provide a scientific basis for the development of more effective cancer treatment strategies.AIM To explore the role of circulating and infiltrating B cell subsets in the immune microenvironment of CRC,explore the potential driving mechanism of B cell development,analyze the interaction between B cells and other immune cells in the immune microenvironment and the functions of communication molecules,and search for possible regulatory pathways to promote the anti-tumor effects of B cells.METHODS A total of 69 paracancer(normal),tumor and peripheral blood samples were collected from 23 patients with CRC from The Cancer Genome Atlas database(https://portal.gdc.cancer.gov/).After the immune cells were sorted by multicolor flow cytometry,the single cell transcriptome and B cell receptor group library were sequenced using the 10X Genomics platform,and the data were analyzed using bioinformatics tools such as Seurat.The differences in the number and function of B cell infiltration between tumor and normal tissue,the interaction between B cell subsets and T cells and myeloid cell subsets,and the transcription factor regulatory network of B cell subsets were explored and analyzed.RESULTS Compared with normal tissue,the infiltrating number of CD20+B cell subsets in tumor tissue increased significantly.Among them,germinal center B cells(GCB)played the most prominent role,with positive clone expansion and heavy chain mutation level increasing,and the trend of differentiation into memory B cells increased.However,the number of plasma cells in the tumor microenvironment decreased significantly,and the plasma cells secreting IgA antibodies decreased most obviously.In addition,compared with the immune microenvironment of normal tissues,GCB cells in tumor tissues became more closely connected with other immune cells such as T cells,and communication molecules that positively regulate immune function were significantly enriched.CONCLUSION The role of GCB in CRC tumor microenvironment is greatly enhanced,and its affinity to tumor antigen is enhanced by its significantly increased heavy chain mutation level.Meanwhile,GCB has enhanced its association with immune cells in the microenvironment,which plays a positive anti-tumor effect.展开更多
The application of microorganisms as probiotics is limited due to lack of safety evaluation.Here,a novel multi-stress-tolerant yeast Meyerozyma guilliermondii GXDK6 with aroma-producing properties was identified from ...The application of microorganisms as probiotics is limited due to lack of safety evaluation.Here,a novel multi-stress-tolerant yeast Meyerozyma guilliermondii GXDK6 with aroma-producing properties was identified from marine mangrove microorganisms.Its safety and probiotic properties were assessed in accordance with phenotype and whole-genome sequencing analysis.Results showed that the genes and phenotypic expression of related virulence,antibiotic resistance and retroelement were rarely found.Hyphal morphogenesis genes(SIT4,HOG1,SPA2,ERK1,ICL1,CST20,HSP104,TPS1,and RHO1)and phospholipase secretion gene(VPS4)were annotated.True hyphae and phospholipase were absent.Only one retroelement(Tad1-65_BG)was found.Major biogenic amines(BAs)encoding genes were absent,except for spermidine synthase(JA9_002594),spermine synthase(JA9_004690),and tyrosine decarboxylase(inx).The production of single BAs and total BAs was far below the food-defined thresholds.GXDK6 had no resistance to common antifungal drugs.Virulence enzymes,such as gelatinase,DNase,hemolytic,lecithinase,and thrombin were absent.Acute toxicity test with mice demonstrated that GXDK6 is safe.GXDK6 has a good reproduction ability in the simulation gastrointestinal tract.GXDK6 also has a strong antioxidant ability,β-glucosidase,and inulinase activity.To sum up,GXDK6 is considered as a safe probiotic for human consumption and food fermentation.展开更多
Objective: to explore the value of 16S rRNA sequencing in analyzing the microbial community of diabetic foot skin. Methods: 26 cases of diabetic foot patients admitted to our hospital (March 2019-April 2021) were sele...Objective: to explore the value of 16S rRNA sequencing in analyzing the microbial community of diabetic foot skin. Methods: 26 cases of diabetic foot patients admitted to our hospital (March 2019-April 2021) were selected by random sample sampling method, and their foot skin samples were collected. 16S rRNA sequencing technology was used to identify microbial species and analyze the flora characteristics and the correlation between skin micro-ecological groups and the occurrence of diabetic foot disease was analyzed. Results: the course of diabetes in 17 patients was long and the level of glycosylated hemoglobin was high. The difference between the two methods was significant (P>0.05). Wagner level of diabetic foot patients was positively correlated with the risk of skin microecology imbalance (r=8.256, P<0.001). Conclusion: compared with the culture results, 16S rRNA sequencing analysis of pathogenic bacteria has higher accuracy, simple operation and significant application value, which can be further popularized.展开更多
[Objectives]This study was conducted to detect and analyze tomato leaf curl New Delhi virus(ToLCNDV).[Methods]Through PCR detection,sequence analysis,and pathogenicity verification,tomato leaf curl New Delhi virus(ToL...[Objectives]This study was conducted to detect and analyze tomato leaf curl New Delhi virus(ToLCNDV).[Methods]Through PCR detection,sequence analysis,and pathogenicity verification,tomato leaf curl New Delhi virus(ToLCNDV)was identified in zucchini exhibiting systemic disease symptoms during a 2024 outbreak in Qingzhou City,Shandong Province,and was designated as ToLCNDV-SD.[Results]Specific primer amplification showed that all eight diseased samples produced bands of 504 bp(DNA-A)and 892 bp(DNA-B).Sequencing analysis revealed that ToLCNDV-SD DNA-A shared 96.10%homology with an Indonesian melon isolate(LC421834.1),while DNA-B showed 88.31%homology with a Malaysian bitter gourd isolate(MW248678.1).Phylogenetic analysis indicated its closest relationship with Southeast Asian cucurbit-infecting isolates.Friction transmission tests confirmed that the virus could spread mechanically,inducing typical symptoms 14 d after inoculation with positive PCR detection.[Conclusions]This study provides important insights for understanding the epidemic mechanisms and control strategies of ToLCNDV in China.展开更多
[Objective] The study aimed to investigate the genetic variation characters of entire sequences between two H9N2 subtype avian influenza virus strains and other reference strains.[Method] The entire sequences of 8 gen...[Objective] The study aimed to investigate the genetic variation characters of entire sequences between two H9N2 subtype avian influenza virus strains and other reference strains.[Method] The entire sequences of 8 genes were obtained by using RT-PCR,and these sequences were analyzed with that of six H9N2 subtype avian influenza isolates in homology comparison and genetic evolution relation.[Result] The results showed that the nucleotide sequence of entire gene of the strain shared 91.1%-95.4% homology with other seven reference strains,and PG08 shared the highest homology 91.3% with C/BJ/1/94;ZD06 shared the highest homology 92.3% with D/HK/Y280/97.HA cleavage sites of two H9N2 subtype avian influenza virus isolated strains were PARSSR/GLF,typical of mildly pathogenic avian influenza virus.[Conclusion] Phylogenetic tree for entire gene of eight strains showed that the genetic relationship was the closest between ZD06 and C/Pak/2/99 strains,which belonged to the Eurasian lineage;PG08 shared the highest homology 91.3% with ZD06,it may be the product of gene rearrangements of other sub-lines.展开更多
Oviductus Ranae is the dried oviduct of female Rana tem-poraria chensinensis （David）, distributed mainly in North- eastern China. Oviductus Ranae is one of the best-known and highly valued oriental foods and medicin...Oviductus Ranae is the dried oviduct of female Rana tem-poraria chensinensis （David）, distributed mainly in North- eastern China. Oviductus Ranae is one of the best-known and highly valued oriental foods and medicines. Traditional Chinese medicine holds that Oviductus Ranae can nourish yin, moisten lung and replenish the kidney essence. Meanwhile, activities of Oviductus Ranae such as anti-aging, anti-lipemic, anti-oxidation and anti-fatigue have also been demonstrated by modern phar-macological studies. Previous studies have shown that Oviductus Ranae is mainly composed of proteins, which are up to 50% or more.展开更多
Seed coat color affects the appearance and commodity quality of mung beans(Vigna radiata L.).The substances that affect mung bean seed coat color are mainly flavonoids,which have important medicinal value.Mapping the ...Seed coat color affects the appearance and commodity quality of mung beans(Vigna radiata L.).The substances that affect mung bean seed coat color are mainly flavonoids,which have important medicinal value.Mapping the seed coat color gene in mung beans would facilitate the development of new varieties and improve their value.In this study,an F2 mapping population consisting of 546 plants was constructed using Jilv9(black seed coat)and BIS9805(green seed coat).Using bulk segregated analysis(BSA)sequencing and kompetitive allele-specific PCR(KASP)markers,the candidate region related to seed coat color was finally narrowed to 0.66 Mb on chromosome(Chr.)4 and included eight candidate genes.Combined transcriptome and metabolome analyses showed that three of the eight candidate genes(LOC106758748,LOC106758747,and LOC106759075)were differentially expressed,which may have caused the differences in flavonoid metabolite content between Jilv9 and BIS9805.These findings can provide a research basis for cloning the genes related to seed coat color and accelerate molecular markerassisted selection breeding in mung beans.展开更多
METHODS:In the present study,female SpragueDawley rats were randomly divided into four groups with 6 of each,including normal control group(NC),jumpinginduced muscle injury model group(JI),JI with electroacupuncture s...METHODS:In the present study,female SpragueDawley rats were randomly divided into four groups with 6 of each,including normal control group(NC),jumpinginduced muscle injury model group(JI),JI with electroacupuncture stimulation treatment group(EA),and JI with non-electroacupuncture stimulation group(NEA).Transmission electron microscopy,transcriptome sequencing and analysis,prediction of protein interaction networks,real-time polymerase chain reaction verification,and Western blotting were performed on the gastrocnemius muscle of ipsilateral lower limbs.RESULTS:The structural repair of injured gastrocnemius myofibers following jumping training in EA rats was better than that of NEA rats.A total of 136 genes were differentially expressed in EA rats relative to JI rats,with 55 genes upregulated and 81 genes downregulated.According to results of transcriptome analysis,and prediction of protein mutual interaction by the online STRING database,Heat shock protein beta-7(Hspb7)and myozenin2(Myoz2)genes were targeted.Expressions of Hspb7 and Myoz2 mRNAs were increased in EA rats relative to JI rats(P<0.05).The expression of Hspb7 protein was upregulated in EA rats relative to that in NC,JI,and NEA rats(P<0.01,<0.05,and<0.05,respectively).The expression of Myoz2 protein was upregulated in EA rats relative to that in NC and JI rats(both P<0.01,respectively).CONCLUSIONS:The present results suggest that electroacupuncture stimulation at Zusanli(ST36)could improve muscle healing following jumping-induced muscle injury,owing to the upregulation of Hspb7 and Myoz2 proteins.展开更多
Three isolates of H9N2 Avian Influenza viruses (AIV) were isolated from chickens in Guangxi province. Eight pairs of specific primers were designed and synthesized according to the sequences of H9N2 at GenBank. phylog...Three isolates of H9N2 Avian Influenza viruses (AIV) were isolated from chickens in Guangxi province. Eight pairs of specific primers were designed and synthesized according to the sequences of H9N2 at GenBank. phylogenetic analysis showed a high degree of homology between the Guangxi isolates and isolates from Guangdong and Jiangsu provinces, suggesting that the Guangxi isolates originated from the same source. However, the eight genes of the three isolates from Guangxi were not in the same sublineages in their respective phylogenetic trees, which suggests that they were products of natural reassortment between H9N2 avian influenza viruses from different sublineages. The 9 nucleotides ACAGAGATA which encode amino acids T, G, I were absent between nucleotide 205 and 214 in the open reading frame of the NA gene in the Guangxi isolates. AIV strains that infect human have, in their HA proteins, leucine at position 226. The analysis of deduced amino acid sequence of HA proteins showed that position 226 of these isolates contained glycine instead of leucine, suggesting that these three isolates differ from H9N2 AIV strains isolated from human infections.展开更多
Using double-stranded RNA(dsRNA)technology and sequence-independent amplification(SIA),the molecular identification on infected Rehmannia glutinosa in the field with mosaic symptoms was performed and the whole-genome ...Using double-stranded RNA(dsRNA)technology and sequence-independent amplification(SIA),the molecular identification on infected Rehmannia glutinosa in the field with mosaic symptoms was performed and the whole-genome of the Rehmannia mosaic virus(ReMV)Shanxi isolate(ReMV-SX)was sequenced.Sequencing analysis showed that the virus that infected Rehmannia glutinosa was Rehmannia mosaic virus(ReMV).The full-length of the obtained ReMV-SX sequence(GenBank accession no.JX575184)was 6395 nt,containing four open reading frames(ORFs).The sequence homology analysis of the complete nucleotide sequence showed that ReMV-SX was 93.8%-97.0%homologous to ReMV in Tobamovirus subgroup Ⅰ,while only 49.8%-58.9%homologous to the isolates in subgroups Ⅱ and Ⅲ of the same genus.Phylogenetic analysis showed that ReMV-SX and ReMV-Henan formed a separate branch and had the closest genetic relationship.The results laid the foundation for ongoing researches in the taxonomic status and evolution of ReMV and for further investigating the pathogenic mechanism of ReMV infecting Rehmannia glutinosa.展开更多
The metagenomic DNA of disease tissue samples from four kinds of major edible fungus was extracted by CTAB method combined with DNA gel recovery kit. The genomie DNA was amplified by polymerase chain reaction using th...The metagenomic DNA of disease tissue samples from four kinds of major edible fungus was extracted by CTAB method combined with DNA gel recovery kit. The genomie DNA was amplified by polymerase chain reaction using the universal primers of 16S rDNA and 18S rDNA, and then mone, elonal sequenced after ligated and transformed, rDNA sequences of 20 positive clones were selected randomly from each pair of primers for sequence alignment. The results showed that there were two bacterial diseases and two fungul diseases in the samples, respectively.展开更多
OBJECTIVE: To investigate the mechanism of Qixuan Yijianing(芪玄抑甲宁,QYN) in minimizing cardiac injury in Graves′ disease(GD) mice using micro RNA(mi RNA) sequencing analysis. METHODS: Female BALB/c mice were rando...OBJECTIVE: To investigate the mechanism of Qixuan Yijianing(芪玄抑甲宁,QYN) in minimizing cardiac injury in Graves′ disease(GD) mice using micro RNA(mi RNA) sequencing analysis. METHODS: Female BALB/c mice were randomly divided into the modeling and control groups(CG). The modeling group was established with Ad-TSHR289. Following 10 weeks of successful modeling, the mice were randomly assigned to four groups: model(MG), methimazole(MMI), QYN low-dose(LD), and high-dose(HD). After four weeks of treatment, the heart rate, heart volume, and heart index were measured, and the levels of aspartate aminotransferase(AST), lactate dehydrogenase(LDH), α-hydroxybutyrate dehydrogenase( α-HBD), creatine kinase(CK), and creatine kinase MB isoenzyme(CK-MB) in the serum were detected using a biochemical analyzer. Hematoxylin-eosin and Masson staining were used to determine histological changes in cardiac tissue. The heart tissues in the CG, MG, and HD groups were selected, and mi RNA sequencing was used to identify differentially expressed mi RNAs. A bioinformatics database was used to predict the target genes of differential mi RNAs, and Gene Ontology(GO), and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis were conducted on the predicted target genes. RESULTS: As compared to the CG group, the MG group's heart rate, heart volume, heart index, AST, CK, CK-MB, LDH, α-HBD, myocardial fiber thickness, and collagen fiber significantly increased, all P < 0.01, while following QYN, these indicators improved in the HD group, all P < 0.01 or P < 0.05. Compared to the CG group, the MG group identified 151 differentially expressed mi RNAs, with 42 mi RNAs downregulated and 109 mi RNAs upregulated;compared to the MG group, the HD group identified 70 differentially expressed mi RNAs, 40 were downregulated, and 30 were upregulated. The GO functions of differential mi RNA target genes are mostly enriched in cardiac development regulation, cardiac contraction control, heart rate regulation, and so on. The most enriched KEGG pathways include the mitogenactivated protein kinase, Erb B, Hippo, forkhead box protein O, and Wnt signaling pathways. CONCLUSION: QYN may protect the cardiac structure and function and minimize cardiac damage caused by GD by regulating relevant target genes and signaling pathways through mi RNAs which include mi R-206-3p, mi R-122-5p, and mi R-200a-3p.展开更多
[Objective] The research aimed to carry out the cloning, identification and sequence analysis of full-length cDNA of carp interferon γ-2β （IFNγ-2β）. [Method] The cDNA library of peripheral blood leucocytes which...[Objective] The research aimed to carry out the cloning, identification and sequence analysis of full-length cDNA of carp interferon γ-2β （IFNγ-2β）. [Method] The cDNA library of peripheral blood leucocytes which were separated from carp and stimulated with mitogen was screened by a probe labeled with DIG. The IFNγ- 2β EST sequence was picked out from the constructed cDNA library of peripheral blood leucocyte, and the full length of carp interferon γ-2β was cloned. In addition, the sequence analysis was carried out. [Result] Three positive clones were obtained. Sequence analysis indicated that the sequence had a 119 bp 5’-UTR and a 218 bp 3’-UTR, and the open reading frame （ORF）of this gene was 537 bp which putatively coded 178 amino acids and there were several instable motifs for mRNA （ATTTA） in the 3’-untranslated region. Its homology with IFN from GenBank was up to 97% . Analysis on protein sequence and structure showed that the predicted protein sequence was identified as an IFN family signature. [Conclusion] The research laid the foundation for further studying the expression manner, function characteristic and regulation mechanism of IFNγ-2β in vivo and the action mechanism in the inflammatory reaction, emergency reaction and immune response.展开更多
Platelets act as a crucial indicator for monitoring hypercoagulability and thrombosis and a key target for pharmacological intervention.Genotype-phenotype association studies have confirmed that platelet traits are qu...Platelets act as a crucial indicator for monitoring hypercoagulability and thrombosis and a key target for pharmacological intervention.Genotype-phenotype association studies have confirmed that platelet traits are quantitatively regulated by multiple genes.However,there is currently a lack of genetic studies on the heterogeneity of platelet traits in β-thalassemia under a hypercoagulable state.Here,we studied the phenotypic heterogeneity of platelet count(PLT)and mean platelet volume(MPV)in a cohort of 1020β-thalassemia patients.We further performed a functionally informed whole-genome sequencing(WGS)association analysis of common variants and rare variants for PLT and MPV in 916 patients through integrative analysis of WGS data and functional annotation data.Extreme phenotypic heterogeneity of platelet traits was observed in β-thalassemia patients.Additionally,the common variant-based gene-level analysis identified RNF144B as a novel gene associated with MPV.The rare variant analysis identified several novel associations in both coding and noncoding regions,including missense rare variants of PPP2R5C associated with PLT and missense rare variants of TSSK1B associated with MPV.In conclusion,this comprehensive and systematic whole-genome scan of platelet traits in the β-thalassemia cohort reveals the specific genetic regulation of platelet traits in the context of β-thalassemia,providing potential targets for intervention.展开更多
BACKGROUND The NOD-like receptor thermal protein domain associated protein 3(NLRP3)inflammasome may play an important role in diabetic kidney disease(DKD).However,the exact link remains unclear.AIM To investigate the ...BACKGROUND The NOD-like receptor thermal protein domain associated protein 3(NLRP3)inflammasome may play an important role in diabetic kidney disease(DKD).However,the exact link remains unclear.AIM To investigate the role of the NLRP3 inflammasome in DKD.METHODS Using datasets from the Gene Expression Omnibus database,30 NLRP3 inflammasome-related genes were identified.Differentially expressed genes were selected using differential expression analysis,whereas intersecting genes were selected based on overlapping differentially expressed genes and NLRP3 inflammasome-related genes.Subsequently,three machine learning algorithms were used to screen genes,and biomarkers were identified by overlapping the genes from the three algorithms.Potential biomarkers were validated by western blotting in a db/db mouse model of diabetes.RESULTS Two biomarkers,sirtuin 2(SIRT2)and caspase 1(CASP1),involved in the Leishmania infection pathway were identified.Both biomarkers were expressed in endothelial cells.Pseudo-temporal analysis based on endothelial cells showed that DKD mostly occurs during the mid-differentiation stage.Western blotting results showed that CASP1 expression was higher in the DKD group than in the control group(P<0.05),and SIRT2 content decreased(P<0.05).CONCLUSION SIRT2 and CASP1 provide a potential theoretical basis for DKD treatment.展开更多
[ Objective ] The aim of this study is to clone and analyze the actin gene from Rehmannia glutinosa. [ Method ] Degenerate primers were designed according to the conserved regions of actin sequences of Rehmannia gluti...[ Objective ] The aim of this study is to clone and analyze the actin gene from Rehmannia glutinosa. [ Method ] Degenerate primers were designed according to the conserved regions of actin sequences of Rehmannia glutinosa and its similar species, RT-PCR was next conducted to amplify the actin gene from Rehmannia glutinosa. [ Result] The amplified fragment is 724 bp and correspondingly 240 amino acids. The BLAST results indicate that the homology between the amplified fragment and other higher plants for aetin gene sequences and amino acid are more than 80% and 90%, respectively, suggesting that the amplified fragment is the actin gene of Rehmannia glutinosa. [ Conclusion] Phylogenetic analysis shows that the actin gene of Rehmannia glutinosa has an intimate genetic relationship with actin7 gene of Nicotiana tabacum.展开更多
[ Objective] The aim of this study was to provide a theoretical basis for exploring the major genes affecting intramuscular fat （IMF） deposition. [Method] Taking 383 pigs from five breeds including Mashen Pig, Large...[ Objective] The aim of this study was to provide a theoretical basis for exploring the major genes affecting intramuscular fat （IMF） deposition. [Method] Taking 383 pigs from five breeds including Mashen Pig, Large White Pig, Landrace, Duroc and Shanxi White Pig as the experimental animals, polymorphisms of partial fragments in the third intron of porcine H-FABP gene were detected by PCR-SSCP method, and then the polymorphic fragments were sequenced. [ Result] Two alleles, designated as A and B, were found at the locus 346 in the third intron of porcine H-FABP gene, and the mutation was caused by a A→G substitution. [ Conclusion] A polymorphic locus was discovered in the third intron of porcine H-FABP gene in this experiment, laying a foundation for the further study on the relationship between H-FABP gene and IMF content.展开更多
[Objective] cDNA from squalene synthase was cloned and sequenced.[Method] A pair of specific primers was designed according to the cDNA gene sequence of squalene synthase published in GenBank.Total RNA was extracted f...[Objective] cDNA from squalene synthase was cloned and sequenced.[Method] A pair of specific primers was designed according to the cDNA gene sequence of squalene synthase published in GenBank.Total RNA was extracted from the cell of Artemisia apiacea.The genes of squalene synthase were amplified by using RT-PCR.It was connected with pMD19-T vector and the cloned fragment sequences were analyzed.[Result] SS gene with the whole length of 1 257 bp was amplified and the fragment encoded 418 amino acids.The homo...展开更多
[ Objective] The aim was to analyze the reason and epidemic trend of PRRSV, and provide theoretical basis for preventing and controlling PRRS. [Methed]According to the sequence of ATCC VR-2332 strain porcine reproduct...[ Objective] The aim was to analyze the reason and epidemic trend of PRRSV, and provide theoretical basis for preventing and controlling PRRS. [Methed]According to the sequence of ATCC VR-2332 strain porcine reproductive and respiratory syndrome virus published by the GenBank, the primers were designed and synthesized. ORF5 gene sequences of seven prevalence strains were amplified by RT-PCR. The sequences of ORF5 genes were analyzed by DNAStar and compared with those of ATCC VR-2332, CH-1 a, B J-4, LV-M96262 and MLV vaccine strains, phylogenetic tree among isolates was analyzed. [Result] Analysis of nucleotide sequence showed that the homology was 88.1% - 98.8%, 89.9% -95.2%, 85.6% -98.7% between ORF5 genes of seven prevalence strains and VR-2332, CH-1a, BJ-4, and the homology was 54.7% -56.9% between ORF5 genes and LV. Analysis of amino acid sequence showed that the homology was 88.1% -96.8%, 88.1% - 94.5%, 86.1% -96.5% between ORF5 genes of seven prevalence strains and VR-2332, CH-1a, bBJ-4, the homology was 54.7% -56.2% between the ORF5 genes and LV.[ Conclusion] The variation of prevalence strains was great in the ORF5 gene region, the homology of ORF5 gene sequence was higher and genetic relationship was nearer during prevalence strains in the same region, or was far in different regions.展开更多
基金supported by Qingdao Key Medical and Health Discipline ProjectThe Intramural Research Program of the Affiliated Hospital of Qingdao University,No. 4910Qingdao West Coast New Area Science and Technology Project,No. 2020-55 (all to SW)。
文摘Border-associated macrophages are located at the interface between the brain and the periphery, including the perivascular spaces, choroid plexus, and meninges. Until recently, the functions of border-associated macrophages have been poorly understood and largely overlooked. However, a recent study reported that border-associated macrophages participate in stroke-induced inflammation, although many details and the underlying mechanisms remain unclear. In this study, we performed a comprehensive single-cell analysis of mouse border-associated macrophages using sequencing data obtained from the Gene Expression Omnibus(GEO) database(GSE174574 and GSE225948). Differentially expressed genes were identified, and enrichment analysis was performed to identify the transcription profile of border-associated macrophages. CellChat analysis was conducted to determine the cell communication network of border-associated macrophages. Transcription factors were predicted using the ‘pySCENIC' tool. We found that, in response to hypoxia, borderassociated macrophages underwent dynamic transcriptional changes and participated in the regulation of inflammatory-related pathways. Notably, the tumor necrosis factor pathway was activated by border-associated macrophages following ischemic stroke. The pySCENIC analysis indicated that the activity of signal transducer and activator of transcription 3(Stat3) was obviously upregulated in stroke, suggesting that Stat3 inhibition may be a promising strategy for treating border-associated macrophages-induced neuroinflammation. Finally, we constructed an animal model to investigate the effects of border-associated macrophages depletion following a stroke. Treatment with liposomes containing clodronate significantly reduced infarct volume in the animals and improved neurological scores compared with untreated animals. Taken together, our results demonstrate comprehensive changes in border-associated macrophages following a stroke, providing a theoretical basis for targeting border-associated macrophages-induced neuroinflammation in stroke treatment.
文摘BACKGROUND The complexity of the immune microenvironment has an impact on the treatment of colorectal cancer(CRC),one of the most prevalent malignancies worldwide.In this study,multi-omics and single-cell sequencing techniques were used to investigate the mechanism of action of circulating and infiltrating B cells in CRC.By revealing the heterogeneity and functional differences of B cells in cancer immunity,we aim to deepen our understanding of immune regulation and provide a scientific basis for the development of more effective cancer treatment strategies.AIM To explore the role of circulating and infiltrating B cell subsets in the immune microenvironment of CRC,explore the potential driving mechanism of B cell development,analyze the interaction between B cells and other immune cells in the immune microenvironment and the functions of communication molecules,and search for possible regulatory pathways to promote the anti-tumor effects of B cells.METHODS A total of 69 paracancer(normal),tumor and peripheral blood samples were collected from 23 patients with CRC from The Cancer Genome Atlas database(https://portal.gdc.cancer.gov/).After the immune cells were sorted by multicolor flow cytometry,the single cell transcriptome and B cell receptor group library were sequenced using the 10X Genomics platform,and the data were analyzed using bioinformatics tools such as Seurat.The differences in the number and function of B cell infiltration between tumor and normal tissue,the interaction between B cell subsets and T cells and myeloid cell subsets,and the transcription factor regulatory network of B cell subsets were explored and analyzed.RESULTS Compared with normal tissue,the infiltrating number of CD20+B cell subsets in tumor tissue increased significantly.Among them,germinal center B cells(GCB)played the most prominent role,with positive clone expansion and heavy chain mutation level increasing,and the trend of differentiation into memory B cells increased.However,the number of plasma cells in the tumor microenvironment decreased significantly,and the plasma cells secreting IgA antibodies decreased most obviously.In addition,compared with the immune microenvironment of normal tissues,GCB cells in tumor tissues became more closely connected with other immune cells such as T cells,and communication molecules that positively regulate immune function were significantly enriched.CONCLUSION The role of GCB in CRC tumor microenvironment is greatly enhanced,and its affinity to tumor antigen is enhanced by its significantly increased heavy chain mutation level.Meanwhile,GCB has enhanced its association with immune cells in the microenvironment,which plays a positive anti-tumor effect.
基金This research was supported by the Funding Project of Chinese Central Government Guiding to the Guangxi Local Science and Technology Development(GUIKEZY21195021)the Natural Science Fund for Distinguished Young Scholars of Guangxi Zhuang Autonomous Region of China(2019GXNSFFA245011)+3 种基金the Funding Project of Chinese Central Government Guiding to the Nanning Local Science and Technology Development(20231012)the Funding Projects of Guangxi Key Research and Development Plan(GUIKE AB23075173)the Funding Project of Technological Development from Angel Yeast(Chongzuo)Co.,Ltd.(JS1006020230722019)the Innovation Project of Guangxi Graduate Education(YCBZ2021012).
文摘The application of microorganisms as probiotics is limited due to lack of safety evaluation.Here,a novel multi-stress-tolerant yeast Meyerozyma guilliermondii GXDK6 with aroma-producing properties was identified from marine mangrove microorganisms.Its safety and probiotic properties were assessed in accordance with phenotype and whole-genome sequencing analysis.Results showed that the genes and phenotypic expression of related virulence,antibiotic resistance and retroelement were rarely found.Hyphal morphogenesis genes(SIT4,HOG1,SPA2,ERK1,ICL1,CST20,HSP104,TPS1,and RHO1)and phospholipase secretion gene(VPS4)were annotated.True hyphae and phospholipase were absent.Only one retroelement(Tad1-65_BG)was found.Major biogenic amines(BAs)encoding genes were absent,except for spermidine synthase(JA9_002594),spermine synthase(JA9_004690),and tyrosine decarboxylase(inx).The production of single BAs and total BAs was far below the food-defined thresholds.GXDK6 had no resistance to common antifungal drugs.Virulence enzymes,such as gelatinase,DNase,hemolytic,lecithinase,and thrombin were absent.Acute toxicity test with mice demonstrated that GXDK6 is safe.GXDK6 has a good reproduction ability in the simulation gastrointestinal tract.GXDK6 also has a strong antioxidant ability,β-glucosidase,and inulinase activity.To sum up,GXDK6 is considered as a safe probiotic for human consumption and food fermentation.
文摘Objective: to explore the value of 16S rRNA sequencing in analyzing the microbial community of diabetic foot skin. Methods: 26 cases of diabetic foot patients admitted to our hospital (March 2019-April 2021) were selected by random sample sampling method, and their foot skin samples were collected. 16S rRNA sequencing technology was used to identify microbial species and analyze the flora characteristics and the correlation between skin micro-ecological groups and the occurrence of diabetic foot disease was analyzed. Results: the course of diabetes in 17 patients was long and the level of glycosylated hemoglobin was high. The difference between the two methods was significant (P>0.05). Wagner level of diabetic foot patients was positively correlated with the risk of skin microecology imbalance (r=8.256, P<0.001). Conclusion: compared with the culture results, 16S rRNA sequencing analysis of pathogenic bacteria has higher accuracy, simple operation and significant application value, which can be further popularized.
基金Supported by Taishan Industry Leading Talent Program in Shandong Province(tscx202306156)Weifang Science and Technology Development Program(2024GX073).
文摘[Objectives]This study was conducted to detect and analyze tomato leaf curl New Delhi virus(ToLCNDV).[Methods]Through PCR detection,sequence analysis,and pathogenicity verification,tomato leaf curl New Delhi virus(ToLCNDV)was identified in zucchini exhibiting systemic disease symptoms during a 2024 outbreak in Qingzhou City,Shandong Province,and was designated as ToLCNDV-SD.[Results]Specific primer amplification showed that all eight diseased samples produced bands of 504 bp(DNA-A)and 892 bp(DNA-B).Sequencing analysis revealed that ToLCNDV-SD DNA-A shared 96.10%homology with an Indonesian melon isolate(LC421834.1),while DNA-B showed 88.31%homology with a Malaysian bitter gourd isolate(MW248678.1).Phylogenetic analysis indicated its closest relationship with Southeast Asian cucurbit-infecting isolates.Friction transmission tests confirmed that the virus could spread mechanically,inducing typical symptoms 14 d after inoculation with positive PCR detection.[Conclusions]This study provides important insights for understanding the epidemic mechanisms and control strategies of ToLCNDV in China.
基金Supported by a Sub-project of 973 Program of China(2005CB523001)~~
文摘[Objective] The study aimed to investigate the genetic variation characters of entire sequences between two H9N2 subtype avian influenza virus strains and other reference strains.[Method] The entire sequences of 8 genes were obtained by using RT-PCR,and these sequences were analyzed with that of six H9N2 subtype avian influenza isolates in homology comparison and genetic evolution relation.[Result] The results showed that the nucleotide sequence of entire gene of the strain shared 91.1%-95.4% homology with other seven reference strains,and PG08 shared the highest homology 91.3% with C/BJ/1/94;ZD06 shared the highest homology 92.3% with D/HK/Y280/97.HA cleavage sites of two H9N2 subtype avian influenza virus isolated strains were PARSSR/GLF,typical of mildly pathogenic avian influenza virus.[Conclusion] Phylogenetic tree for entire gene of eight strains showed that the genetic relationship was the closest between ZD06 and C/Pak/2/99 strains,which belonged to the Eurasian lineage;PG08 shared the highest homology 91.3% with ZD06,it may be the product of gene rearrangements of other sub-lines.
基金supported by the National Key Technology Research and Development Program of the Ministry of Science and Technology of China (No. 2011BAI03B00)the National Science and Technology Major Project of the Ministry of Science and Technology of China (No. 2011ZX09401-305)
文摘Oviductus Ranae is the dried oviduct of female Rana tem-poraria chensinensis （David）, distributed mainly in North- eastern China. Oviductus Ranae is one of the best-known and highly valued oriental foods and medicines. Traditional Chinese medicine holds that Oviductus Ranae can nourish yin, moisten lung and replenish the kidney essence. Meanwhile, activities of Oviductus Ranae such as anti-aging, anti-lipemic, anti-oxidation and anti-fatigue have also been demonstrated by modern phar-macological studies. Previous studies have shown that Oviductus Ranae is mainly composed of proteins, which are up to 50% or more.
基金supported by the National Natural Science Foundation of China(32301928)the Basic Research Program of Shanxi Province,China(20210302124504)+3 种基金the China Agriculture Research System of MOF and MARA-Food Legumes(CARS08-G10)the National Laboratory Project of Coarse Grain Germplasm Resources Innovation and Molecular Breeding,China(K462202040-01)the Ph D of Shanxi Agricultural University Scientific Research Start-up Project,China(2021BQ43)the Scientific Research Project of Shanxi Agricultural University,China(YZGC098)。
文摘Seed coat color affects the appearance and commodity quality of mung beans(Vigna radiata L.).The substances that affect mung bean seed coat color are mainly flavonoids,which have important medicinal value.Mapping the seed coat color gene in mung beans would facilitate the development of new varieties and improve their value.In this study,an F2 mapping population consisting of 546 plants was constructed using Jilv9(black seed coat)and BIS9805(green seed coat).Using bulk segregated analysis(BSA)sequencing and kompetitive allele-specific PCR(KASP)markers,the candidate region related to seed coat color was finally narrowed to 0.66 Mb on chromosome(Chr.)4 and included eight candidate genes.Combined transcriptome and metabolome analyses showed that three of the eight candidate genes(LOC106758748,LOC106758747,and LOC106759075)were differentially expressed,which may have caused the differences in flavonoid metabolite content between Jilv9 and BIS9805.These findings can provide a research basis for cloning the genes related to seed coat color and accelerate molecular markerassisted selection breeding in mung beans.
基金Supported by Science and Technology Department of Henan Province Effect Mechanisms of Acupuncture to Promote the Repair of Injured Skeletal Muscle(No.212102310260)Study on the Relationship between Hspb7 Protein and JAK/STAT Pathway and Electroacupuncture Intervention in Muscle Injury based on mRNA-Seq Analysis(No.222102320072)Science and Technology Bureau o Kaifeng:the Effect of Platelet Plasma Injection on Patellar Tendon Fibrosis(No.2003006)。
文摘METHODS:In the present study,female SpragueDawley rats were randomly divided into four groups with 6 of each,including normal control group(NC),jumpinginduced muscle injury model group(JI),JI with electroacupuncture stimulation treatment group(EA),and JI with non-electroacupuncture stimulation group(NEA).Transmission electron microscopy,transcriptome sequencing and analysis,prediction of protein interaction networks,real-time polymerase chain reaction verification,and Western blotting were performed on the gastrocnemius muscle of ipsilateral lower limbs.RESULTS:The structural repair of injured gastrocnemius myofibers following jumping training in EA rats was better than that of NEA rats.A total of 136 genes were differentially expressed in EA rats relative to JI rats,with 55 genes upregulated and 81 genes downregulated.According to results of transcriptome analysis,and prediction of protein mutual interaction by the online STRING database,Heat shock protein beta-7(Hspb7)and myozenin2(Myoz2)genes were targeted.Expressions of Hspb7 and Myoz2 mRNAs were increased in EA rats relative to JI rats(P<0.05).The expression of Hspb7 protein was upregulated in EA rats relative to that in NC,JI,and NEA rats(P<0.01,<0.05,and<0.05,respectively).The expression of Myoz2 protein was upregulated in EA rats relative to that in NC and JI rats(both P<0.01,respectively).CONCLUSIONS:The present results suggest that electroacupuncture stimulation at Zusanli(ST36)could improve muscle healing following jumping-induced muscle injury,owing to the upregulation of Hspb7 and Myoz2 proteins.
基金supported by National Bai Qian Wan Talents Engineering Foudation (Grant No. 9452006-03 )Guangxi Science Technology Bureau (GKG- 0719004-3A)Guangxi Husbandry and Fisheries Bureau.
文摘Three isolates of H9N2 Avian Influenza viruses (AIV) were isolated from chickens in Guangxi province. Eight pairs of specific primers were designed and synthesized according to the sequences of H9N2 at GenBank. phylogenetic analysis showed a high degree of homology between the Guangxi isolates and isolates from Guangdong and Jiangsu provinces, suggesting that the Guangxi isolates originated from the same source. However, the eight genes of the three isolates from Guangxi were not in the same sublineages in their respective phylogenetic trees, which suggests that they were products of natural reassortment between H9N2 avian influenza viruses from different sublineages. The 9 nucleotides ACAGAGATA which encode amino acids T, G, I were absent between nucleotide 205 and 214 in the open reading frame of the NA gene in the Guangxi isolates. AIV strains that infect human have, in their HA proteins, leucine at position 226. The analysis of deduced amino acid sequence of HA proteins showed that position 226 of these isolates contained glycine instead of leucine, suggesting that these three isolates differ from H9N2 AIV strains isolated from human infections.
基金Supported by the National Natural Science Foundation of China(31772130)China Agriculture Research System(CARS-21)。
文摘Using double-stranded RNA(dsRNA)technology and sequence-independent amplification(SIA),the molecular identification on infected Rehmannia glutinosa in the field with mosaic symptoms was performed and the whole-genome of the Rehmannia mosaic virus(ReMV)Shanxi isolate(ReMV-SX)was sequenced.Sequencing analysis showed that the virus that infected Rehmannia glutinosa was Rehmannia mosaic virus(ReMV).The full-length of the obtained ReMV-SX sequence(GenBank accession no.JX575184)was 6395 nt,containing four open reading frames(ORFs).The sequence homology analysis of the complete nucleotide sequence showed that ReMV-SX was 93.8%-97.0%homologous to ReMV in Tobamovirus subgroup Ⅰ,while only 49.8%-58.9%homologous to the isolates in subgroups Ⅱ and Ⅲ of the same genus.Phylogenetic analysis showed that ReMV-SX and ReMV-Henan formed a separate branch and had the closest genetic relationship.The results laid the foundation for ongoing researches in the taxonomic status and evolution of ReMV and for further investigating the pathogenic mechanism of ReMV infecting Rehmannia glutinosa.
基金Supported by Project for State Edible Fungus Industrial Technology System Construction(CARS-24)Agricultural Improved Variety Project of Shandong Province(2012LZ006-04)Science and Technology Development Program of Shandong Province
文摘The metagenomic DNA of disease tissue samples from four kinds of major edible fungus was extracted by CTAB method combined with DNA gel recovery kit. The genomie DNA was amplified by polymerase chain reaction using the universal primers of 16S rDNA and 18S rDNA, and then mone, elonal sequenced after ligated and transformed, rDNA sequences of 20 positive clones were selected randomly from each pair of primers for sequence alignment. The results showed that there were two bacterial diseases and two fungul diseases in the samples, respectively.
基金Research Grant from the National Natural Science Foundation of China:Mechanism Study on the Treatment of Graves'Disease with Yiqi Yangyin Qinggan Sanjie Method based on T helper cell 17 and Related Factors (No. 81302898)Mudanjiang Medical University Research Initiation Fund Project Funding:Mechanism Study on the Treatment of Graves'Disease with Yiqi Yangyin Qinggan Sanjie Method Based on Thyroid Angiogenesis (No.2023-MYBSKY-008)Funding for the Mudanjiang City Applied Technology Research and Development Program Project:Metabolomics study of extract of Herb of Common Leibnitzia based on Ultra Performance Liquid Chromatography-Time of Flight Mass Spectrometry Technology on Collagen-induced Arthritis Rats (No. HT2020NS093)。
文摘OBJECTIVE: To investigate the mechanism of Qixuan Yijianing(芪玄抑甲宁,QYN) in minimizing cardiac injury in Graves′ disease(GD) mice using micro RNA(mi RNA) sequencing analysis. METHODS: Female BALB/c mice were randomly divided into the modeling and control groups(CG). The modeling group was established with Ad-TSHR289. Following 10 weeks of successful modeling, the mice were randomly assigned to four groups: model(MG), methimazole(MMI), QYN low-dose(LD), and high-dose(HD). After four weeks of treatment, the heart rate, heart volume, and heart index were measured, and the levels of aspartate aminotransferase(AST), lactate dehydrogenase(LDH), α-hydroxybutyrate dehydrogenase( α-HBD), creatine kinase(CK), and creatine kinase MB isoenzyme(CK-MB) in the serum were detected using a biochemical analyzer. Hematoxylin-eosin and Masson staining were used to determine histological changes in cardiac tissue. The heart tissues in the CG, MG, and HD groups were selected, and mi RNA sequencing was used to identify differentially expressed mi RNAs. A bioinformatics database was used to predict the target genes of differential mi RNAs, and Gene Ontology(GO), and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis were conducted on the predicted target genes. RESULTS: As compared to the CG group, the MG group's heart rate, heart volume, heart index, AST, CK, CK-MB, LDH, α-HBD, myocardial fiber thickness, and collagen fiber significantly increased, all P < 0.01, while following QYN, these indicators improved in the HD group, all P < 0.01 or P < 0.05. Compared to the CG group, the MG group identified 151 differentially expressed mi RNAs, with 42 mi RNAs downregulated and 109 mi RNAs upregulated;compared to the MG group, the HD group identified 70 differentially expressed mi RNAs, 40 were downregulated, and 30 were upregulated. The GO functions of differential mi RNA target genes are mostly enriched in cardiac development regulation, cardiac contraction control, heart rate regulation, and so on. The most enriched KEGG pathways include the mitogenactivated protein kinase, Erb B, Hippo, forkhead box protein O, and Wnt signaling pathways. CONCLUSION: QYN may protect the cardiac structure and function and minimize cardiac damage caused by GD by regulating relevant target genes and signaling pathways through mi RNAs which include mi R-206-3p, mi R-122-5p, and mi R-200a-3p.
基金Supported by the National Natural Science Foundation of China (30972277)the Fundamental Research Fund of Jilin University (200903250)~~
文摘[Objective] The research aimed to carry out the cloning, identification and sequence analysis of full-length cDNA of carp interferon γ-2β （IFNγ-2β）. [Method] The cDNA library of peripheral blood leucocytes which were separated from carp and stimulated with mitogen was screened by a probe labeled with DIG. The IFNγ- 2β EST sequence was picked out from the constructed cDNA library of peripheral blood leucocyte, and the full length of carp interferon γ-2β was cloned. In addition, the sequence analysis was carried out. [Result] Three positive clones were obtained. Sequence analysis indicated that the sequence had a 119 bp 5’-UTR and a 218 bp 3’-UTR, and the open reading frame （ORF）of this gene was 537 bp which putatively coded 178 amino acids and there were several instable motifs for mRNA （ATTTA） in the 3’-untranslated region. Its homology with IFN from GenBank was up to 97% . Analysis on protein sequence and structure showed that the predicted protein sequence was identified as an IFN family signature. [Conclusion] The research laid the foundation for further studying the expression manner, function characteristic and regulation mechanism of IFNγ-2β in vivo and the action mechanism in the inflammatory reaction, emergency reaction and immune response.
基金supported by research funding from the National Natural Science Foundation of China(Grant No.U20A20353)the Guangdong Basic and Applied Basic Research Foundation(Grant No.2023A1515110850)+1 种基金the China Postdoctoral Science Foundation(Grant No.2023M741585)the Medical Scientific Research Foundation of Guangdong Province(Grant No.A2022443),China.
文摘Platelets act as a crucial indicator for monitoring hypercoagulability and thrombosis and a key target for pharmacological intervention.Genotype-phenotype association studies have confirmed that platelet traits are quantitatively regulated by multiple genes.However,there is currently a lack of genetic studies on the heterogeneity of platelet traits in β-thalassemia under a hypercoagulable state.Here,we studied the phenotypic heterogeneity of platelet count(PLT)and mean platelet volume(MPV)in a cohort of 1020β-thalassemia patients.We further performed a functionally informed whole-genome sequencing(WGS)association analysis of common variants and rare variants for PLT and MPV in 916 patients through integrative analysis of WGS data and functional annotation data.Extreme phenotypic heterogeneity of platelet traits was observed in β-thalassemia patients.Additionally,the common variant-based gene-level analysis identified RNF144B as a novel gene associated with MPV.The rare variant analysis identified several novel associations in both coding and noncoding regions,including missense rare variants of PPP2R5C associated with PLT and missense rare variants of TSSK1B associated with MPV.In conclusion,this comprehensive and systematic whole-genome scan of platelet traits in the β-thalassemia cohort reveals the specific genetic regulation of platelet traits in the context of β-thalassemia,providing potential targets for intervention.
基金Joint Funds for the Innovation of Science and Technology,Fujian Province,No.2021Y9106Fujian Provincial Health Technology Project,No.2021GGA033the Natural Science Foundation of Fujian Province,No.2024J011234.
文摘BACKGROUND The NOD-like receptor thermal protein domain associated protein 3(NLRP3)inflammasome may play an important role in diabetic kidney disease(DKD).However,the exact link remains unclear.AIM To investigate the role of the NLRP3 inflammasome in DKD.METHODS Using datasets from the Gene Expression Omnibus database,30 NLRP3 inflammasome-related genes were identified.Differentially expressed genes were selected using differential expression analysis,whereas intersecting genes were selected based on overlapping differentially expressed genes and NLRP3 inflammasome-related genes.Subsequently,three machine learning algorithms were used to screen genes,and biomarkers were identified by overlapping the genes from the three algorithms.Potential biomarkers were validated by western blotting in a db/db mouse model of diabetes.RESULTS Two biomarkers,sirtuin 2(SIRT2)and caspase 1(CASP1),involved in the Leishmania infection pathway were identified.Both biomarkers were expressed in endothelial cells.Pseudo-temporal analysis based on endothelial cells showed that DKD mostly occurs during the mid-differentiation stage.Western blotting results showed that CASP1 expression was higher in the DKD group than in the control group(P<0.05),and SIRT2 content decreased(P<0.05).CONCLUSION SIRT2 and CASP1 provide a potential theoretical basis for DKD treatment.
基金National Natural Science Foundation of China (No 30472155)Beijing Natural Science Foundation (No 5062035)~~
文摘[ Objective ] The aim of this study is to clone and analyze the actin gene from Rehmannia glutinosa. [ Method ] Degenerate primers were designed according to the conserved regions of actin sequences of Rehmannia glutinosa and its similar species, RT-PCR was next conducted to amplify the actin gene from Rehmannia glutinosa. [ Result] The amplified fragment is 724 bp and correspondingly 240 amino acids. The BLAST results indicate that the homology between the amplified fragment and other higher plants for aetin gene sequences and amino acid are more than 80% and 90%, respectively, suggesting that the amplified fragment is the actin gene of Rehmannia glutinosa. [ Conclusion] Phylogenetic analysis shows that the actin gene of Rehmannia glutinosa has an intimate genetic relationship with actin7 gene of Nicotiana tabacum.
基金Supported by Doctoral Start-up Fund for Scientific Research in North China Coal Medical University (07101168)~~
文摘[ Objective] The aim of this study was to provide a theoretical basis for exploring the major genes affecting intramuscular fat （IMF） deposition. [Method] Taking 383 pigs from five breeds including Mashen Pig, Large White Pig, Landrace, Duroc and Shanxi White Pig as the experimental animals, polymorphisms of partial fragments in the third intron of porcine H-FABP gene were detected by PCR-SSCP method, and then the polymorphic fragments were sequenced. [ Result] Two alleles, designated as A and B, were found at the locus 346 in the third intron of porcine H-FABP gene, and the mutation was caused by a A→G substitution. [ Conclusion] A polymorphic locus was discovered in the third intron of porcine H-FABP gene in this experiment, laying a foundation for the further study on the relationship between H-FABP gene and IMF content.
文摘[Objective] cDNA from squalene synthase was cloned and sequenced.[Method] A pair of specific primers was designed according to the cDNA gene sequence of squalene synthase published in GenBank.Total RNA was extracted from the cell of Artemisia apiacea.The genes of squalene synthase were amplified by using RT-PCR.It was connected with pMD19-T vector and the cloned fragment sequences were analyzed.[Result] SS gene with the whole length of 1 257 bp was amplified and the fragment encoded 418 amino acids.The homo...
文摘[ Objective] The aim was to analyze the reason and epidemic trend of PRRSV, and provide theoretical basis for preventing and controlling PRRS. [Methed]According to the sequence of ATCC VR-2332 strain porcine reproductive and respiratory syndrome virus published by the GenBank, the primers were designed and synthesized. ORF5 gene sequences of seven prevalence strains were amplified by RT-PCR. The sequences of ORF5 genes were analyzed by DNAStar and compared with those of ATCC VR-2332, CH-1 a, B J-4, LV-M96262 and MLV vaccine strains, phylogenetic tree among isolates was analyzed. [Result] Analysis of nucleotide sequence showed that the homology was 88.1% - 98.8%, 89.9% -95.2%, 85.6% -98.7% between ORF5 genes of seven prevalence strains and VR-2332, CH-1a, BJ-4, and the homology was 54.7% -56.9% between ORF5 genes and LV. Analysis of amino acid sequence showed that the homology was 88.1% -96.8%, 88.1% - 94.5%, 86.1% -96.5% between ORF5 genes of seven prevalence strains and VR-2332, CH-1a, bBJ-4, the homology was 54.7% -56.2% between the ORF5 genes and LV.[ Conclusion] The variation of prevalence strains was great in the ORF5 gene region, the homology of ORF5 gene sequence was higher and genetic relationship was nearer during prevalence strains in the same region, or was far in different regions.
