This study aims to evaluate the preventive effect of the exogenous protein chIFN-γ-chCD154 against Salmonella typhimurium(S.typhimurium)infection in White Leghorn chick-ens,and the potential mechanism.In this study,Escherichia coli was used to express the proteins chIFN-γ,chCD154 and chIFN-γ-chCD154.Before S.typhimurium infection,the White Leghorn chickens were pre-immunized via drinking water for three consecutive days,and infected with S.typhimurium by gavage.The results from Western blot,quantitative real-time PCR(qRT-PCR)analysis and histopathology analysis showed that compared to chIFN-γ and chCD154,chIFN-γ-chCD154 pre-treatment could synergistically increase the survival rate of infected chickens,reduce the bacterial load in the liver and cecum,and attenuate the pathological damage of liver and cecum.Moreover,chIFN-γ-chCD154 significantly decreased the mRNA expression of Toll-like receptor-4(TLR-4)and myeloid differentiation primary response gene 88(MyD88)in the cecum,and then inhibited the mRNA expression of NF-κB and pro-inflammatory cytokines(TNF-α and IL-6),and maintained the integrity of the intestinal tight junction proteins(zo-1,claudin-1,occlu-din).Compared with single protein pretreatment,chIFN-γ-chCD154 pretreatment significantly up-regulated the mRNA expression of the genes related to the vitamin D(VD)pathway(cyp27b1,VD receptor VDR,antimicrobial peptide AvBD7 and cathelicidin-b1)in S.typhimurium-infected peripheral blood mononuclear cells(PBMCs).The results of colony counting showed that the num-ber of S.typhimurium in the chIFN-γ-chCD154 group were the lowest.Also,chIFN-γ-chCD154 could up-regulate the relative abundance of beneficial bacteria such as Blautia,Ruminococcus,En-terococcus and Faecalibacterium,while down-regulate the relative abundance of harmful bacteria such as Staphylococcaceae in the cecum and improve the intestinal dysbiosis.In conclusion,chIFN-γ-chCD154 could activate the VD-antimicrobial peptide pathway and inhibit the TLR-4/MyD88/NF-κB inflammatory signaling pathway in S.typhimurium-infected chickens,which significantly improve the intestinal barrier function,reduce the inflammatory damage of liver and cecum,im-prove the structure of cecum microbial,promote the health of intestinal tract,and provide theoreti-cal basis for the development of chIFN-γ-chCD154 as a safe and effective alternative antibiotic.

To investigate the distribution and biological characteristics of Trueperella pyogenes in caprine mastitis,67 milk samples from were collected from goats with mastitis in 9 dairy goat farms in the Guanzhong area of Shaanxi Province.A total of 21 strains of T.pyogenes were isolated through bacterial separation and culture.PCR amplification was conducted for 8 virulence genes of the 21 isolated strains,and the susceptibility of the strains to 10 antimicrobial drugs was tested u-sing the micro broth dilution method.Nine selected strains were subjected to genomic sequencing using Illumina Nova Seq 6000 next-generation sequencing technology,and their pathogenicity was verified by inoculating mice through the teat canal.The results of virulence gene detection showed that the positive rates of plo,cbpA,nanH,nanP,fimA,fimC-,and fimE genes among the 21 iso-lated strains were 100.0％,38.1％,52.6％,31.6％,61.9％,57.1％,and 52.4％,respectively.In con-trast,the fimG gene was not detected,which was consistent with the results of genomic compari-son by next-generation sequencing.The drug susceptibility test results showed that the susceptibil-ity rates of the isolated strains to clindamycin(CLI),ceftiofur(CEF),and vancomycin(VAN)were 100.0％,while the resistance rates to penicillin(PEN),erythromycin(ERY),sulfamethox-azole-trimethoprim(SXT),florfenicol(FFC),tetracycline(TE),gentamicin(GEN),and enroflox-acin(ENR)were 76.2％,52.1％,52.4％,52.4％,47.6％,42.9％,and 33.3％,respectively.The pro-portion of strains resistant to at least three antimicrobial drugs was 71.4％(15/21).ResFinder a-nalysis of the resistant genes identified 8 types of resistance genes in the genomes of the 9 strains,including ant(2")-Ia and ant(3")-Ia mediating aminoglycoside resistance,cmlA1 and cmx media-ting chloramphenicol resistance,erm(X)and lnu(A)mediating macrolide resistance,sul1 media-ting sulfonamide resistance,and tet(W)mediating tetracycline resistance.Except for tet(W),cm-lA,and erm(X),the other resistance genes were detected for the first time in T.pyogenes in Chi-na.In animal pathogenicity tests,mice inoculated with the isolated strains showed signs of mastitis such as redness,congestion,and inflammatory cell infiltration in the glandular tissue within 24 h,indicating that the isolated strains have pathogenicity to mouse mammary tissue.This study pro-vides preliminary insights into the biological characteristics and pathogenicity of T.pyogenes from caprine mastitis,enriching the data on its drug resistance and genomics,and offering reference and guidance for the prevention and treatment of T.pyogenes infections in large-scale dairy goat farm-ing.

This study aims to evaluate the preventive effect of the exogenous protein chIFN-γ-chCD154 against Salmonella typhimurium(S.typhimurium)infection in White Leghorn chick-ens,and the potential mechanism.In this study,Escherichia coli was used to express the proteins chIFN-γ,chCD154 and chIFN-γ-chCD154.Before S.typhimurium infection,the White Leghorn chickens were pre-immunized via drinking water for three consecutive days,and infected with S.typhimurium by gavage.The results from Western blot,quantitative real-time PCR(qRT-PCR)analysis and histopathology analysis showed that compared to chIFN-γ and chCD154,chIFN-γ-chCD154 pre-treatment could synergistically increase the survival rate of infected chickens,reduce the bacterial load in the liver and cecum,and attenuate the pathological damage of liver and cecum.Moreover,chIFN-γ-chCD154 significantly decreased the mRNA expression of Toll-like receptor-4(TLR-4)and myeloid differentiation primary response gene 88(MyD88)in the cecum,and then inhibited the mRNA expression of NF-κB and pro-inflammatory cytokines(TNF-α and IL-6),and maintained the integrity of the intestinal tight junction proteins(zo-1,claudin-1,occlu-din).Compared with single protein pretreatment,chIFN-γ-chCD154 pretreatment significantly up-regulated the mRNA expression of the genes related to the vitamin D(VD)pathway(cyp27b1,VD receptor VDR,antimicrobial peptide AvBD7 and cathelicidin-b1)in S.typhimurium-infected peripheral blood mononuclear cells(PBMCs).The results of colony counting showed that the num-ber of S.typhimurium in the chIFN-γ-chCD154 group were the lowest.Also,chIFN-γ-chCD154 could up-regulate the relative abundance of beneficial bacteria such as Blautia,Ruminococcus,En-terococcus and Faecalibacterium,while down-regulate the relative abundance of harmful bacteria such as Staphylococcaceae in the cecum and improve the intestinal dysbiosis.In conclusion,chIFN-γ-chCD154 could activate the VD-antimicrobial peptide pathway and inhibit the TLR-4/MyD88/NF-κB inflammatory signaling pathway in S.typhimurium-infected chickens,which significantly improve the intestinal barrier function,reduce the inflammatory damage of liver and cecum,im-prove the structure of cecum microbial,promote the health of intestinal tract,and provide theoreti-cal basis for the development of chIFN-γ-chCD154 as a safe and effective alternative antibiotic.

To investigate the distribution and biological characteristics of Trueperella pyogenes in caprine mastitis,67 milk samples from were collected from goats with mastitis in 9 dairy goat farms in the Guanzhong area of Shaanxi Province.A total of 21 strains of T.pyogenes were isolated through bacterial separation and culture.PCR amplification was conducted for 8 virulence genes of the 21 isolated strains,and the susceptibility of the strains to 10 antimicrobial drugs was tested u-sing the micro broth dilution method.Nine selected strains were subjected to genomic sequencing using Illumina Nova Seq 6000 next-generation sequencing technology,and their pathogenicity was verified by inoculating mice through the teat canal.The results of virulence gene detection showed that the positive rates of plo,cbpA,nanH,nanP,fimA,fimC-,and fimE genes among the 21 iso-lated strains were 100.0％,38.1％,52.6％,31.6％,61.9％,57.1％,and 52.4％,respectively.In con-trast,the fimG gene was not detected,which was consistent with the results of genomic compari-son by next-generation sequencing.The drug susceptibility test results showed that the susceptibil-ity rates of the isolated strains to clindamycin(CLI),ceftiofur(CEF),and vancomycin(VAN)were 100.0％,while the resistance rates to penicillin(PEN),erythromycin(ERY),sulfamethox-azole-trimethoprim(SXT),florfenicol(FFC),tetracycline(TE),gentamicin(GEN),and enroflox-acin(ENR)were 76.2％,52.1％,52.4％,52.4％,47.6％,42.9％,and 33.3％,respectively.The pro-portion of strains resistant to at least three antimicrobial drugs was 71.4％(15/21).ResFinder a-nalysis of the resistant genes identified 8 types of resistance genes in the genomes of the 9 strains,including ant(2")-Ia and ant(3")-Ia mediating aminoglycoside resistance,cmlA1 and cmx media-ting chloramphenicol resistance,erm(X)and lnu(A)mediating macrolide resistance,sul1 media-ting sulfonamide resistance,and tet(W)mediating tetracycline resistance.Except for tet(W),cm-lA,and erm(X),the other resistance genes were detected for the first time in T.pyogenes in Chi-na.In animal pathogenicity tests,mice inoculated with the isolated strains showed signs of mastitis such as redness,congestion,and inflammatory cell infiltration in the glandular tissue within 24 h,indicating that the isolated strains have pathogenicity to mouse mammary tissue.This study pro-vides preliminary insights into the biological characteristics and pathogenicity of T.pyogenes from caprine mastitis,enriching the data on its drug resistance and genomics,and offering reference and guidance for the prevention and treatment of T.pyogenes infections in large-scale dairy goat farm-ing.

Objective To observe the clinical effectiveness of the experience of famous traditional Chinese medicine doctors in the treatment of residual sleep disturbances in patients remitted from major depressive disorder.Methods The 108 patients remitted from major depressive disorder with residual sleep disturbances who were treated in the department of Psychosomatic Medicine,Beijing Hospital of Traditional Chinese Medicine Affiliated to Capital Medical University from June 2021 to December 2023,were randomly divided into acupuncture group(acupuncture treatments consisted of 24 sessions duration over 8 weeks)and medication group(patients take estazolam 1 mg daily for 8 weeks),54 cases in each.Assessments were carried out at baseline,4,8,12 and 20 weeks,including the Pittsburgh Sleep Quality Index(PSQI),Insomnia Severity Index(ISI),Hamilton Depression Scale(HAMD),Hamilton Anxiety Scale(HAMA),and SF-36.Results The difference in PSQI,ISI,HAMD,HAMA,and mental health scores(P<0.05)in the acupuncture group was greater than in the medication group.The total effective rate of the acupuncture and medication groups was 72.22%and 14.81%,respectively,and the difference was statistically significant(P<0.05).Conclusion Acupuncture can obviously improve sleep quality,alleviate symptoms of depression and anxiety,and improve quality of life in patients with major depressive disorder.

Objective To evaluate the clinical efficacy and safety of acupuncture in the treatment of grand anxiety disorder by applying Meta-analysis.Methods Randomized controlled trials on acupuncture treatment of generalized anxiety disorder in 7 Chinese and English databases from the establishment of the database to May 1,2022 were searched for screening,inclusion and literature risk assessment.Meta-analysis and subgroup analysis of different outcome indicators and/or intervention measures were performed using Stata 16 and Review Manager 5.4 software.Results Finally,20 randomized controlled trials were included,including 1661 cases and 4 intervention measures(acupuncture+electroacupuncture,acupuncture,electroacupuncture and western medicine).The results of meta-analysis indicated that there were statistically significant differences in HAMA and SAS scores between acupuncture group and western medicine group(P<0.05).The results of subgroup analysis showed that there was statistically significant difference in Hamilton anxiety scale(HAMA)score between acupuncture group and alprazolam.There was a statistically significant difference in self-rating anxiety scale(SAS)score between acupuncture group and simple paroxetine,buspirone,chlorhexidine/oryzanol/propranolol.In the evaluation of side effect scale,the difference between acupuncture(including electroacupuncture)group and western medicine group was statistically significant(P<0.01).Conclusion Acupuncture(including electroacupuncture)can effectively improve anxiety and symptoms with efficacy no less than that of Western medicine,and has the advantages of fewer adverse effects and higher safety.However,due to the quantity and quality of the included literature,multi-center,large sample,and high-quality randomized controlled studies are still needed to further corroborate the above conclusions.

Objective To observe the clinical effectiveness of the experience of famous traditional Chinese medicine doctors in the treatment of residual sleep disturbances in patients remitted from major depressive disorder.Methods The 108 patients remitted from major depressive disorder with residual sleep disturbances who were treated in the department of Psychosomatic Medicine,Beijing Hospital of Traditional Chinese Medicine Affiliated to Capital Medical University from June 2021 to December 2023,were randomly divided into acupuncture group(acupuncture treatments consisted of 24 sessions duration over 8 weeks)and medication group(patients take estazolam 1 mg daily for 8 weeks),54 cases in each.Assessments were carried out at baseline,4,8,12 and 20 weeks,including the Pittsburgh Sleep Quality Index(PSQI),Insomnia Severity Index(ISI),Hamilton Depression Scale(HAMD),Hamilton Anxiety Scale(HAMA),and SF-36.Results The difference in PSQI,ISI,HAMD,HAMA,and mental health scores(P<0.05)in the acupuncture group was greater than in the medication group.The total effective rate of the acupuncture and medication groups was 72.22%and 14.81%,respectively,and the difference was statistically significant(P<0.05).Conclusion Acupuncture can obviously improve sleep quality,alleviate symptoms of depression and anxiety,and improve quality of life in patients with major depressive disorder.

INTRODUCTIONThe current metastatic category (M) of nasopharyngeal carcinoma (NPC) is a "catch-all" classification, covering a heterogeneous group of tumors ranging from potentially curable to incurable. The aim of this study was to design an M categorization system that could be applied in planning the treatment of NPC with synchronous metastasis.

METHODSA total of 505 NPC patients diagnosed with synchronous metastasis at Sun Yat-sen University Cancer Center between 2000 and 2009 were involved. The associations of clinical variables, metastatic features, and a proposed M categorization system with overall survival (OS) were determined by using Cox regression model.

RESULTSMultivariate analysis showed that Union for International Cancer Control (UICC) N category (N1-3/N0), number of metastatic lesions (multiple/single), liver involvement (yes/no), radiotherapy to primary tumor (yes/no), and cycles of chemotherapy (>4/≤4) were independent prognostic factors for OS. We defined the following subcategories based on liver involvement and the number of metastatic lesions: M1a, single lesion confined to an isolated organ or location except the liver; M1b, single lesion in the liver and/or multiple lesions in any organs or locations except the liver; and M1c, multiple lesions in the liver. Of the 505 cases, 74 (14.7%) were classified as M1a, 296 (58.6%) as M1b, 134 (26.5%) as M1c, and 1 was not specified. The three M1 subcategories showed significant difference in OS [M1b vs. M1a, hazard ratio (HR) = 1.69, 95% confidence interval (CI) = 1.16-2.48, P = 0.007; M1c vs. M1a, HR = 2.64, 95% CI = 1.75-3.98, P < 0.001].

CONCLUSIONSWe developed an M categorization system based on the independent factors related to the prognosis of patients with metastatic NPC. This system may be helpful to further optimize individualized care for NPC patients.