This article systematically explores the traditional Chinese medicine （TCM） pathogenesis evolution and metastatic mechanisms of advanced gastric cancer from the perspective of "phlegm-toxin flowing infusion". Based on this theory， it is proposed that advanced gastric cancer presents a pathogenesis evolution characterized by healthy qi depletion， phlegm-toxin flowing infusion， and collateral damage with blood stasis. Healthy qi depletion is the root of the disease， in which spleen and kidney insufficiency leads to weakened healthy qi and inability to resist pathogenic factors. Phlegm-toxin flowing infusion is the core pathogenesis， where phlegm-dampness and cancer toxin intertwine and migrate to other organs along with qi movement. Collateral damage with blood stasis constitutes the final pathological stage of metastasis， where phlegm-toxin obstruction damages the collaterals， and congealed qi and blood form accumulations. On this basis， the meridian pathways and metastasis mechanisms of advanced gastric cancer spreading to multiple sites such as the liver， peritoneum， lungs， bones， brain， ovary， and lymph nodes are further analyzed， providing references for clinical management and research of this disease.

To construct a recombinant fowl adenovirus 4(FAdV-4)expressing the Cap protein of goose astrovirus genotype 2(GoAstV-2),the expression cassette of Cap gene was inserted into the natural 1 966 bp deletion region of the FAdV-4 genome in the infectious clone p15A-cm-FAdV4-HNJZ.The resulted recombinant plasmid p15A-cm-FAdV4-HNJZ-Cap/GoAstV-2 was linearized with restriction enzyme and transfected into chicken hepatoma cell line(LMH)to rescue the recombinant FAdV-4 expressing the Cap protein of GoAstV-2,rF Ad V4-Cap/GoAstV-2.After 15 passages in LMH cells,the recombinant rFAdV4-Cap/GoAstV-2 was identified by PCR using primers flanking the insertion site of the Cap gene expression cassette and using viral genome DNA extracted from rFAdV4-Cap/GoAstV-2 infected LMH cells as template.LMH cells were in-fected with 15th passage rFAdV4-Cap/GoAstV-2 and indirect immunofluorescence was performed with a polyclonal antibody against Cap protein as the primary antibody.Western blot was carried out with lysates of rFAdV4-Cap/GoAstV-2 infected LMH cells.The in vitro replication dynamic of the 15th passage of the rFAdV4-Cap/GoAstV-2 was also investigated in LMH cells.The results demonstrated that the Cap gene of GoAstV-2 was presented in the genome of the recombinant vi-rus rF AdV4-Cap/Go Ast V-2,and could be expressed stably.The prepared recombinant virus in this study will lay a foundation for developing inactivated bivalent vaccine candidate against co-in-fection of FAdV-4 and GoAstV-2 in goose.

In order to understand the epidemic situation of European porcine reproductive and re-spiratory syndrome virus 1(PRRSV1)in Henan Province,molecular epidemiological investigation and virus identification were conducted on clinical specimens from some large-scale pig farms in Henan Province.RT-PCR detection and sequencing were carried out using specific primers for PRRSVI ORF5 gene.As a result,eight PRRSV1 positive samples were detected with six ORF5 gene sequences and one full gene sequence obtained.A strain of PRRSVI named as HENZMD-10 was successfully isolated using PAM cells.Based on the genetic variation analysis,the six ORF5 genes obtained were all PRRSVI.The isolated strains have relatively large ORF5 gene genetic vari-ation,belonging to different branches on the genetic evolution tree.Among them,HENJZ-11,HENJY-7,and HENJY-8 were relatively close in genetic evolution and belonged to the same branch.The genome length of HENZMD-10 isolate was 15 071 bp.Alignment analysis showed that HENZMD-10 strain shared an 89.1％of nucleotide sequence with the LV strain;62.1％and 61.5％with PRRSV2 ATCC-VR2332 strain and the American NADC30 strain,respectively.The nucleotide similarity of HENZMD-10 strain with the domestic JXA1 strain was 61.6％.The phylo-genetic analysis using complete genome sequence showed that HENZMD-10 strain was relatively close to the domestic isolated PRRSV1 NVDC-NM1-2011,LNEU12 and FJEU13 strains.The suc-cessful isolation of one PRRSV1 strain in Henan Province provides the basis for understanding the epidemic dynamics and prevention and control of PRRSV1 in China.

This study aims to screen the diagnostic biomarkers for fecal antigen of Echinococcus granulosus(E.granulosus)in dogs with high specificity and sensitivity.The sheep-derived EgPSC artificially infected dogs were collected,and the negative and positive fecal samples of dogs with E.granulosus were prepared by arecoline hydrobromide leakage method.Polyclonal antibody,negative fecal antigen-polyclonal antibody conjugates and positive fecal antigen-polyclonal antibody conju-gates were purified by ammonium sulfate precipitation and affinity chromatography,three groups of samples were detected by ELISA and Western blot,LC-MS/MS and bioinformatics analysis were performed on the three groups of samples.The positive fecal antigen-polyclonal antibody con-jugate was used as the treatment group,the polyclonal antibody and the negative fecal antigen-polyclonal antibody conjugates were used as the control groups to screen the unique peptides of the treatment group.ELISA and Western blot showed that only the positive fecal antigen-polyclonal antibody conjugates were positive.According to LC-MS/MS and bioinformatics analysis,11 unique peptides were screened out only in the treatment group.Among them,3 proteins were related to E.granulosus,namely dysferlin,integrator complex 9 and diagnostic antigen gp50,which were mem-brane-associated proteins,INT complex components and diagnostic antigens.This study has pre-liminarily screened out three candidate canine E.granulosus fecal antigen diagnostic markers,pro-viding a reference for further exploration of diagnostic standards for E.granulosus,screening of echinococcosis target genes,and vaccine development.

This study aims to screen the diagnostic biomarkers for fecal antigen of Echinococcus granulosus(E.granulosus)in dogs with high specificity and sensitivity.The sheep-derived EgPSC artificially infected dogs were collected,and the negative and positive fecal samples of dogs with E.granulosus were prepared by arecoline hydrobromide leakage method.Polyclonal antibody,negative fecal antigen-polyclonal antibody conjugates and positive fecal antigen-polyclonal antibody conju-gates were purified by ammonium sulfate precipitation and affinity chromatography,three groups of samples were detected by ELISA and Western blot,LC-MS/MS and bioinformatics analysis were performed on the three groups of samples.The positive fecal antigen-polyclonal antibody con-jugate was used as the treatment group,the polyclonal antibody and the negative fecal antigen-polyclonal antibody conjugates were used as the control groups to screen the unique peptides of the treatment group.ELISA and Western blot showed that only the positive fecal antigen-polyclonal antibody conjugates were positive.According to LC-MS/MS and bioinformatics analysis,11 unique peptides were screened out only in the treatment group.Among them,3 proteins were related to E.granulosus,namely dysferlin,integrator complex 9 and diagnostic antigen gp50,which were mem-brane-associated proteins,INT complex components and diagnostic antigens.This study has pre-liminarily screened out three candidate canine E.granulosus fecal antigen diagnostic markers,pro-viding a reference for further exploration of diagnostic standards for E.granulosus,screening of echinococcosis target genes,and vaccine development.

Objective To investigate the effects of three types of twin-block(TB)appliances on the stress and displacement of anterior teeth,periodontal ligaments,and alveolar bone.Methods A three-dimensional(3D)finite element model was constructed,including maxillofacial bones,articular discs,teeth,and periodontal ligaments.Three types of twin-block appliances were designed:classic twin-block(classic-TB),twin-block with acrylic capping(capping-TB),and clear twin-block aligner(CTBA).All appliances had an inclination angle of 70°,and a masticatory force of 200 N was applied to their inclined planes.The finite element method was used to analyze the stress distribution and displacement differences of anterior teeth.Results All three types of TB appliances induced lingual tilting of maxillary anterior teeth and labial tilting of mandibular anterior teeth.The CTBA group showed the greatest lingual displacement and stress of maxillary anterior teeth,with a maximum stress of 30.6 MPa,while the mandibular anterior teeth in this group exhibited the smallest labial displacement(approximately 0.02 mm)and stress.Additionally,the CTBA group had the lowest compressive stress in mandibular anterior teeth,periodontal ligaments,and alveolar bone,whereas the classic-TB group had the highest.Conclusions In the treatment of Angle Class Ⅱ malocclusion,classic-TB(with or without acrylic capping)causes labial inclination of mandibular anterior teeth.Compared with classic-TB,CTBA effectively reduces the compressive stress and displacement of mandibular anterior teeth,potentially minimizing adverse periodontal risks.However,attention should be paid to the lingual displacement of maxillary anterior teeth.

Objective To investigate the effects of three types of twin-block(TB)appliances on the stress and displacement of anterior teeth,periodontal ligaments,and alveolar bone.Methods A three-dimensional(3D)finite element model was constructed,including maxillofacial bones,articular discs,teeth,and periodontal ligaments.Three types of twin-block appliances were designed:classic twin-block(classic-TB),twin-block with acrylic capping(capping-TB),and clear twin-block aligner(CTBA).All appliances had an inclination angle of 70°,and a masticatory force of 200 N was applied to their inclined planes.The finite element method was used to analyze the stress distribution and displacement differences of anterior teeth.Results All three types of TB appliances induced lingual tilting of maxillary anterior teeth and labial tilting of mandibular anterior teeth.The CTBA group showed the greatest lingual displacement and stress of maxillary anterior teeth,with a maximum stress of 30.6 MPa,while the mandibular anterior teeth in this group exhibited the smallest labial displacement(approximately 0.02 mm)and stress.Additionally,the CTBA group had the lowest compressive stress in mandibular anterior teeth,periodontal ligaments,and alveolar bone,whereas the classic-TB group had the highest.Conclusions In the treatment of Angle Class Ⅱ malocclusion,classic-TB(with or without acrylic capping)causes labial inclination of mandibular anterior teeth.Compared with classic-TB,CTBA effectively reduces the compressive stress and displacement of mandibular anterior teeth,potentially minimizing adverse periodontal risks.However,attention should be paid to the lingual displacement of maxillary anterior teeth.

Objective: To explore the causal relationship between gut microbiota and upper urinary tract stones using Mendelian randomization (MR) analysis，and to investigate the potential mediating role of plasma metabolites. Methods: Data on gut microbiota，plasma metabolites，and upper urinary tract stones were obtained from publicly available Genome-wide Association Studies (GWAS).Bidirectional MR analysis was performed to examine the causal relationship between gut microbiota and upper urinary tract stones.Subsequently，a two-step MR approach was employed to determine whether gut microbiota contribute to upper urinary tract stones through plasma metabolites，and the mediating effects and mediator ratio were calculated.The inverse variance weighted (IVW) method was used as the primary analytical tool，supplemented by Bayesian weighted Mendelian randomization (BWMR)，MR-Egger，and weighted median (WM) analyses.Horizontal pleiotropy and heterogeneity tests were conducted to ensure the robustness of the findings. Results: Bidirectional MR analysis identified causal associations between 7 gut microbial taxa and 6 microbial metabolic pathways with upper urinary tract stones，while the development of upper urinary tract stones affected 13 gut microbial taxa and 5 metabolic pathways.Additionally，43 plasma metabolites (including 27 identified metabolites，8 unidentified metabolites，and 8 metabolite ratios) were causally associated with upper urinary tract stones.The two-step MR analysis identified 11 potential causal pathways.After metabolic pathways and unidentified metabolites were excluded，a causal link mediated by Bacteroides faecis between galactarate and upper urinary tract stones was confirmed，with a mediation proportion of 16.99% (95%CI：5.76%-33.95%，P=0.0371). Conclusion: This study establishes a causal relationship between parabacteroides and upper urinary tract stones，and elucidates the mediating role of galactarate，offering new insights into the pathogenesis and prevention strategies for upper urinary tract stones.

AIM: To investigate the predictive value of serum histone deacetylase 1(HDAC1)and endothelial nitric oxide synthase(eNOS)for infectious endophthalmitis after cataract surgery.METHODS: A total of 362 cataract patients(362 eyes)admitted to our hospital from January 2020 to January 2023 were selected as the research objects. According to the occurrence of postoperative infectious endophthalmitis, they were divided into infection group(15 cases, 15 eyes)and non-infection group(347 cases, 347 eyes). Enzyme linked immunosorbent assay(ELISA)was applied to detect the levels of serum HDAC1 and eNOS in all subjects, and the levels of serum HDAC1 and eNOS in both groups were compared; the influencing factors of infectious endophthalmitis were analyzed by multivariate Logistic regression; the receiver operative curve(ROC)was applied to analyze the predictive value of serum HDAC1 and eNOS levels for postoperative infectious endophthalmitis in cataract patients.RESULTS: The levels of serum HDAC1 and eNOS in the infected group were obviously higher than those in the uninfected group(all P<0.01). Surgical time, vitreous overflow, HDAC1, and eNOS were all risk factors for postoperative infectious endophthalmitis(all P<0.05). ROC results showed that the AUC of HDAC1 and eNOS in predicting postoperative infectious endophthalmitis in cataract patients was 0.878 and 0.877, respectively, with sensitivity of 88.7% and 87.7%, specificity of 70.4% and 7.8%, respectively, while the AUC of the two combination in predicting postoperative infectious endophthalmitis in cataract patients was 0.978, with a sensitivity of 86.7% and a specificity of 85.3%.CONCLUSION:The serum levels of HDAC1 and eNOS in patients with infectious endophthalmitis after cataract surgery are obviously increased, and the combined detection of serum HDAC1 and eNOS can improve the predictive efficacy of infectious endophthalmitis in cataract patients after surgery. Both can provide reference for clinical diagnosis and treatment.

AIM: To investigate the predictive value of serum histone deacetylase 1(HDAC1)and endothelial nitric oxide synthase(eNOS)for infectious endophthalmitis after cataract surgery.METHODS: A total of 362 cataract patients(362 eyes)admitted to our hospital from January 2020 to January 2023 were selected as the research objects. According to the occurrence of postoperative infectious endophthalmitis, they were divided into infection group(15 cases, 15 eyes)and non-infection group(347 cases, 347 eyes). Enzyme linked immunosorbent assay(ELISA)was applied to detect the levels of serum HDAC1 and eNOS in all subjects, and the levels of serum HDAC1 and eNOS in both groups were compared; the influencing factors of infectious endophthalmitis were analyzed by multivariate Logistic regression; the receiver operative curve(ROC)was applied to analyze the predictive value of serum HDAC1 and eNOS levels for postoperative infectious endophthalmitis in cataract patients.RESULTS: The levels of serum HDAC1 and eNOS in the infected group were obviously higher than those in the uninfected group(all P<0.01). Surgical time, vitreous overflow, HDAC1, and eNOS were all risk factors for postoperative infectious endophthalmitis(all P<0.05). ROC results showed that the AUC of HDAC1 and eNOS in predicting postoperative infectious endophthalmitis in cataract patients was 0.878 and 0.877, respectively, with sensitivity of 88.7% and 87.7%, specificity of 70.4% and 7.8%, respectively, while the AUC of the two combination in predicting postoperative infectious endophthalmitis in cataract patients was 0.978, with a sensitivity of 86.7% and a specificity of 85.3%.CONCLUSION:The serum levels of HDAC1 and eNOS in patients with infectious endophthalmitis after cataract surgery are obviously increased, and the combined detection of serum HDAC1 and eNOS can improve the predictive efficacy of infectious endophthalmitis in cataract patients after surgery. Both can provide reference for clinical diagnosis and treatment.