Objective:To analyze the genetic characteristics of varicella-zoster virus（VZV）in Anhui province from 2022 to 2023.Methods:Vesicle fluid and throat swab samples were collected from suspected varicella patients in Anhui province during 2022—2023. Fresh vesicle fluid samples were selected for VZV isolation，and real-time PCR was used for VZV nucleic acid detection. For positive samples，the region containing five single nucleotide polymorphism（SNP）sites in the open reading frame 22（ORF22）fragment was amplified，and PCR products were sequenced to identify viral genotypes. Reference sequences of VZV genotypes were downloaded from GenBank，sequence alignment and phylogenetic tree analysis were performed using Sequencher and MEGA5.0 software. Additionally，four SNP sites in ORF38 and ORF62 fragments were detected to distinguish vaccine strains from wild strains.Results:Among 96 samples from suspected varicella cases，55 of 61 vesicle fluid samples and 21 of 35 throat swab samples were positive for VZV nucleic acid. The virus isolation rate for vesicle fluid samples was 14.75%. Genetic sequencing was successful for 51 strains，all of which were wild strains belonging to the clade 2 genetic branch. Compared with the reference strain of clade 2，the nucleotide and amino acid homologies of the ORF22 fragment were 99.46%~100% and 98.39%~100%，respectively. One strain（2023VZVCZ45）exhibited an A→G mutation at site 37916.Conclusion:The prevalent VZV strains detected in Anhui province during 2022—2023 were all wild strains of clade 2，with no vaccine-associated cases identified.

OBJECTIVE: To investigate the effectiveness of delayed replantation of degloving skin preserved at 4℃ in treatment of limb degloving injuries. METHODS: Between October 2020 and October 2023, 12 patients with limb degloving injuries were admitted. All patients had severe associated injuries or poor wound conditions that prevented primary replantation. There were 7 males and 5 females; age ranged from 29 to 46 years, with an average of 39.2 years. The causes of injury included machine entanglement in 6 cases, traffic accidents in 5 cases, and sharp instrument cuts in 1 case. Time from injury to hospital admission was 0.5-3.0 hours, with an average of 1.3 hours. Injury sites included upper limbs in 7 cases and lower limbs in 5 cases. The range of degloving skin was from 5 cm×4 cm to 15 cm×8 cm, and all degloving skins were intact. The degloving skin was preserved at 4℃. After the patient's vital signs became stable and the wound conditions improved, it was trimmed into medium-thickness skin grafts for replantation. The degloving skin was preserved for 3 to 7 days. At 4 weeks after replantation, the viability of the degloving skin grafts was assessed, including color, elasticity, and sensation of pain. The Vancouver Scar Scale (VSS) was used to assess the scars of the skin grafts during follow-up. RESULTS: At 4 weeks after replantation, 8 cases of skin grafts completely survived and the color was similar with normal skin, with a survival rate of 66.67%. The elasticity of skin grafts (R0 value) ranged from 0.09 to 0.85, with an average of 0.55; moderate pain was reported in 4 cases, mild pain in 3 cases, and no pain in 5 cases. All patients were followed up 12 months. Over time, the VSS scores of all 12 patients gradually decreased, with a range of 4-11 at 12 months (mean, 6.8). CONCLUSION For limb degloving injuries that cannot be replanted immediately and do not have the conditions for deep low-temperature freezing preservation, the method of preserving the degloving skin at 4℃ for delayed replantation can be chosen.
Humans ; Male ; Adult ; Replantation/methods* ; Female ; Degloving Injuries/surgery* ; Middle Aged ; Skin Transplantation/methods* ; Treatment Outcome ; Extremities/injuries* ; Time Factors ; Skin/injuries* ; Tissue Preservation/methods*

Currently, the immunization program has entered a new era of high-quality development. Implementing the standardized management of vaccination units is a necessary means to achieve the high-quality development of immunization programs. Although significant achievements have been made in vaccination work in China, there are still many urgent problems to be solved in vaccination work under the new situation, which poses new challenges to standardized management of vaccination units. This paper analyzes the problems faced by the standardized management of vaccination units based on a review of the work content at various stages of management, and discusses strategies and suggestions for the standardized management of vaccination units.

Objective To explore the biosynthesis of Dipsacus asper Wall.ex Henry triterpenoid saponin,and the UGT gene in Dipsacus asper Wall.ex Henry has been analyzed by the identification of whole genome,genome and prokaryotic expression.Methods The laboratory self-tested sequenced protein sequence files of the Dipsacus asper Wall.ex Henry genome were used.To validate the conserved domains of the sequence of the Dipsacus asper Wall.ex Henry UGT gene,BLASTP and hmmsearch were utilized.Prot-Param,SOMPA,MAGA7.0,Tbtools and other tools were used to investigate the protein physicochemical properties,protein structure,and covariance analysis of the Dipsacus asper Wall.ex Henry UGT gene family,and using the joint analysis of transcriptomic data and metabolomics data,two glycosyltransferases that might be related to triterpene saponin biosynthesis were screened,and expression vectors were constructed for prokaryotic expression.Results 44 Dipsacus asper Wall.ex Henry UGT genes were identified from the Dipsacus asper Wall.ex Henry genome.The length of Dipsacus asper Wall.ex Henry UGT proteins ranged from 49 to 1083 amino acids,with an average molecular weight of 24.86 kDa and an isoelectric point of 4.31-8.59.Dipsacus asper Wall.ex Henry UGT gene family was distributed on eight chromosomes.The phylogenetic tree constructed from the sequences of Dipsacus asper Wall.ex Henry,Arabidopsis thaliana and identified UGTs showed that glycosyltransferase gene families in Dipsacus asper Wall.ex Henry were mainly in the UGT73,UGT81,UGT85,and UGT80 families.Cis-acting element analysis showed that light-responsive elements were the most prevalent elements in the promoter regions of UGT gene family members.Two glycosyltransferases potentially related to triterpene saponin biosynthesis were screened using combined transcriptomics and metabolomics analysis,and were successfully expressed in prokaryotic form.Conclusion In this study,two candidate genes related to the biosynthesis of Dipsacus asper Wall.ex Henry triterpenoid saponins were jointly screened for prokaryotic expression using multi-omics,and were subjected to prokaryotic expression for further validation of the function of the genes.

This study constructed a porcine reproductive and respiratory syndrome virus(PRRSV)NADC30-like strain GP3 recombinant adenovirus vector vaccine through in vitro homologous re-combination to explore its immunological efficacy evaluation at the mouse level.Using type 5 ade-novirus as a vector,a recombinant adenovirus expressing PRRSV GP3 protein was prepared and i-dentified in vitro by fluorescence observation,PCR,and Western blot analysis.Immunize mice with recombinant adenovirus and detect humoral and cellular immune responses induced by recombi-nant adenovirus using indirect ELISA and ELISpot methods.The recombinant adenovirus rAdGP3 was identified by enzyme digestion,PCR,fluorescence and Western blot,indicating that the recom-binant adenovirus rAdGP3 was successfully constructed and packaged.After immunizing mice,spe-cific antibodies and neutralizing antibodies were produced,indicating that the recombinant adenovi-rus could elicit strong humoral immunity.ELISpot and lymphocyte proliferation assays showed that the recombinant adenovirus vaccine could stimulate the secretion of IFN-γ-specific T lymphocytes and induce the proliferation of lymphocytes,indicating that the recombinant adenovi-rus could enhance the level of cellular immune response.In this study,rAdGP3 recombinant adeno-virus was successfully constructed and had good immunogenicity at the mouse level,which provid-ed a reference for the development of novel PRRSV vaccines.

Objective To explore the biosynthesis of Dipsacus asper Wall.ex Henry triterpenoid saponin,and the UGT gene in Dipsacus asper Wall.ex Henry has been analyzed by the identification of whole genome,genome and prokaryotic expression.Methods The laboratory self-tested sequenced protein sequence files of the Dipsacus asper Wall.ex Henry genome were used.To validate the conserved domains of the sequence of the Dipsacus asper Wall.ex Henry UGT gene,BLASTP and hmmsearch were utilized.Prot-Param,SOMPA,MAGA7.0,Tbtools and other tools were used to investigate the protein physicochemical properties,protein structure,and covariance analysis of the Dipsacus asper Wall.ex Henry UGT gene family,and using the joint analysis of transcriptomic data and metabolomics data,two glycosyltransferases that might be related to triterpene saponin biosynthesis were screened,and expression vectors were constructed for prokaryotic expression.Results 44 Dipsacus asper Wall.ex Henry UGT genes were identified from the Dipsacus asper Wall.ex Henry genome.The length of Dipsacus asper Wall.ex Henry UGT proteins ranged from 49 to 1083 amino acids,with an average molecular weight of 24.86 kDa and an isoelectric point of 4.31-8.59.Dipsacus asper Wall.ex Henry UGT gene family was distributed on eight chromosomes.The phylogenetic tree constructed from the sequences of Dipsacus asper Wall.ex Henry,Arabidopsis thaliana and identified UGTs showed that glycosyltransferase gene families in Dipsacus asper Wall.ex Henry were mainly in the UGT73,UGT81,UGT85,and UGT80 families.Cis-acting element analysis showed that light-responsive elements were the most prevalent elements in the promoter regions of UGT gene family members.Two glycosyltransferases potentially related to triterpene saponin biosynthesis were screened using combined transcriptomics and metabolomics analysis,and were successfully expressed in prokaryotic form.Conclusion In this study,two candidate genes related to the biosynthesis of Dipsacus asper Wall.ex Henry triterpenoid saponins were jointly screened for prokaryotic expression using multi-omics,and were subjected to prokaryotic expression for further validation of the function of the genes.

Currently, the immunization program has entered a new era of high-quality development. Implementing the standardized management of vaccination units is a necessary means to achieve the high-quality development of immunization programs. Although significant achievements have been made in vaccination work in China, there are still many urgent problems to be solved in vaccination work under the new situation, which poses new challenges to standardized management of vaccination units. This paper analyzes the problems faced by the standardized management of vaccination units based on a review of the work content at various stages of management, and discusses strategies and suggestions for the standardized management of vaccination units.

This study constructed a porcine reproductive and respiratory syndrome virus(PRRSV)NADC30-like strain GP3 recombinant adenovirus vector vaccine through in vitro homologous re-combination to explore its immunological efficacy evaluation at the mouse level.Using type 5 ade-novirus as a vector,a recombinant adenovirus expressing PRRSV GP3 protein was prepared and i-dentified in vitro by fluorescence observation,PCR,and Western blot analysis.Immunize mice with recombinant adenovirus and detect humoral and cellular immune responses induced by recombi-nant adenovirus using indirect ELISA and ELISpot methods.The recombinant adenovirus rAdGP3 was identified by enzyme digestion,PCR,fluorescence and Western blot,indicating that the recom-binant adenovirus rAdGP3 was successfully constructed and packaged.After immunizing mice,spe-cific antibodies and neutralizing antibodies were produced,indicating that the recombinant adenovi-rus could elicit strong humoral immunity.ELISpot and lymphocyte proliferation assays showed that the recombinant adenovirus vaccine could stimulate the secretion of IFN-γ-specific T lymphocytes and induce the proliferation of lymphocytes,indicating that the recombinant adenovi-rus could enhance the level of cellular immune response.In this study,rAdGP3 recombinant adeno-virus was successfully constructed and had good immunogenicity at the mouse level,which provid-ed a reference for the development of novel PRRSV vaccines.

Objective:To analyze the genetic characteristics of varicella-zoster virus（VZV）in Anhui province from 2022 to 2023.Methods:Vesicle fluid and throat swab samples were collected from suspected varicella patients in Anhui province during 2022—2023. Fresh vesicle fluid samples were selected for VZV isolation，and real-time PCR was used for VZV nucleic acid detection. For positive samples，the region containing five single nucleotide polymorphism（SNP）sites in the open reading frame 22（ORF22）fragment was amplified，and PCR products were sequenced to identify viral genotypes. Reference sequences of VZV genotypes were downloaded from GenBank，sequence alignment and phylogenetic tree analysis were performed using Sequencher and MEGA5.0 software. Additionally，four SNP sites in ORF38 and ORF62 fragments were detected to distinguish vaccine strains from wild strains.Results:Among 96 samples from suspected varicella cases，55 of 61 vesicle fluid samples and 21 of 35 throat swab samples were positive for VZV nucleic acid. The virus isolation rate for vesicle fluid samples was 14.75%. Genetic sequencing was successful for 51 strains，all of which were wild strains belonging to the clade 2 genetic branch. Compared with the reference strain of clade 2，the nucleotide and amino acid homologies of the ORF22 fragment were 99.46%~100% and 98.39%~100%，respectively. One strain（2023VZVCZ45）exhibited an A→G mutation at site 37916.Conclusion:The prevalent VZV strains detected in Anhui province during 2022—2023 were all wild strains of clade 2，with no vaccine-associated cases identified.

In order to explore the role of BspE protein in Brucella infection,yeast two-hybrid tech-nique was used to screen host cell proteins that interact with BspE protein.The constructed BspE recombinant plasmid pGBKT7-BspE was used as bait plasmid to hybridize with the RAW264.7-cD-NA library of mouse mononuclear macrophages by yeast two-hybridization technique.The positive clones were extracted by plasmid,sequenced and co-immunoprecipitation to determine the host cell proteins that could interact with BspE.The subcellular localization of BspE proteins was analyzed by confocal laser microscopy.The physical and chemical properties,protein structure and function of BspE interacting proteins were analyzed by bioinformatics.The siRNA for one of the BspE inter-acting proteins was synthesized,the expression of its gene was silenced in HEK293T cells,and the silenced cells was infected with Brucella M5-90 and the number of intracellular bacteria was coun-ted.The results showed that the decoy plasmid pGBKT7-BspE was successfully constructed,and the plasmid could express BspE protein in yeast.Eight positive clones were obtained from the host cell genome library by yeast two-hybridization.The positive clones were identified as RBM27 and PCBP1 by sequencing,backcross and co-immunoprecipitation.Bioinformatics was used to predict the cell location,protein structure and amino acid composition of RBM27 and PCBP1.After siRNA interference,the expression level of PCBP1 was significantly decreased and the amount of M5-90 in the cell was increased.Brucellosis secreted protein BspE interacts with host proteins RBM27 and PCBPl,and PCBP1 negatively regulates the proliferation of Brucellosis.