Swine enteric coronaviruses(SeCoV),such as porcine epidemic diarrhea virus(PEDV),transmissible gastroenteritis virus(TGEV),and porcine delta coronavirus(PDCoV),cause severe diarrhea in piglets,resulting in substantial losses in pig farming.In this study we establish a triple fluorescence reverse transcription-quantitative PCR(RT-qPCR)method for the simultaneous de-tection of PEDV,TGEV,and PDCoV.The specific primers and probes for each target virus were designed based on conserved sequences from the PEDV M gene,the TGEV ORF 1b gene,and the PDCoV ORF 1b gene respectively.Following the optimization of parameters and conditions,a triple RT-qPCR method was successfully established to simultaneously detect PEDV,TGEV,and PD-CoV.The developed assay exhibits strong specificity for these three pathogens without any cross-reaction with other common porcine viruses like CSFV,PCV2,PoRVA,PRV,and PRRSV.The de-tection limit of linear templates for pTOPO-PEDV 128,pTOPO-TGEV 116,and pTOPO-PDCoV 125 recombinant plasmids were 16.835,17.610 and 17.020 copies/μL,respectively.The intra group and inter group coefficients of variation were less than 5％,with no significant differences observed(P＞0.05).Moreover,the detection consistency rate of the developed RT-qPCR was compared with standard method and showed 100％agreement.Out of 35 small intestine tissue samples,17 tested positive for PEDV,resulting in a positive rate of 48.57％(17/35).The tests for TGEV and PDCoV yielded negative results,and no mixed infections were detected.Based on the above results,the tri-ple RT-qPCR method established is specific,sensitive,stable,and rapid,and can be used for clinical detection and differential diagnosis of PEDV,TGEV,and PDCoV simultaneously,providing a method for the detection and epidemiological investigation of porcine diarrhea coronaviruses.

Objective:To investigate and analyze the surgical method and therapeutic effect of removing benign masses between the nasal radix and eyebrow via a V-shaped cosmetic incision approach through the columella with the assistance of nasal endoscopy.Methods:The data of patients who underwent surgical treatment from May 2020 to August 2023 at the Department of Otorhinolaryngology, Head and Neck Surgery, Linyi People’s Hospital were retrospectively analyzed, because of a mass involving the radix of the nose and the interbrow area. Preoperative CT examination of the nose was performed to clarify the nature, size, and level of the mass, and a V-shaped incision was made in the middle and lower third of the nasal columella, with subcutaneous tunnels along the dorsal median line of the nose, and the mass was excised with the aid of nasal endoscopy. Postoperative follow-up was performed for recurrence of swelling, diplopia, ocular dyskinesia, frontal-facial numbness, and recovery of the operative area; patients were evaluated by visual analogue scoring (VAS) to assess satisfaction with the appearance of the incorporeal scar (0~10 points, higher scores indicate higher satisfaction).Results:A total of 4 patients were included, with an average age of 18.5 years (15-21 years). All 4 patients had complete resection of the tumor through the columellar V-shaped incision approach, and the surgical incisions were all healed by primary intention. Postoperative pathology suggested epidermoid cysts in 1 case, lipoma in 1 case, sebaceous cysts in 1 case, and osteomas in 1 case. Postoperative follow-up ranged from 6 to 24 months, with an average follow-up of 12 months, with no recurrence of the mass, no depression at the radix of the nose and between the eyebrows, and no noticeable scar at the columella. The patients were satisfied with the appearance of the incision scar with a VAS score of 10 points.Conclusion:Nasal endoscopic transnasal columella approach to excision of benign tumors at the root of the nose and between the eyebrows has the advantages of small skin incision and hidden scarring, and has good treatment effect.

Animal experiments are an essential component of life sciences and medical research. However, the external validity and reliability of individual animal studies are frequently challenged by inherent limitations such as small sample sizes, high design heterogeneity, and poor reproducibility, which impede the effective translation of research findings into clinical practice. Systematic reviews and meta-analysis represent a key methodology for integrating existing evidence and enhancing the robustness of conclusions. Currently, however, the application of systematic reviews and meta-analysis in the field of animal experiments lacks standardized guidelines for their conduct and reporting, resulting in inconsistent quality and, to some extent, diminishing their evidence value. To address this issue, this paper aims to systematically delineate the reporting process for systematic reviews and meta-analysis of animal experiments and to propose a set of standardized recommendations that are both scientific and practical. The article's scope encompasses the entire process, from the preliminary preparatory phase [including formulating the population， intervention， comparison and outcome （PICO） question, assessing feasibility, and protocol pre-registration] to the key writing points for each section of the main report. In the core methods section, the paper elaborates on how to implement literature searches, establish eligibility criteria, perform data extraction, and assess the risk of bias, based on the Preferred Reporting Items for Systematic Reviews and Meta-analysis （PRISMA) statement, in conjunction with relevant guidelines and tools such as Animal Research： Reporting of in Vivo Experiments （ARRIVE) and a risk of bias assessment tool developed by the Systematic Review Centre for Laboratory Animal Experimentation (SYRCLE). For the presentation of results, strategies are proposed for clear and transparent display using flow diagrams and tables of characteristics. The discussion section places particular emphasis on how to scientifically interpret pooled effects, thoroughly analyze sources of heterogeneity, evaluate the impact of publication bias, and cautiously discuss the validity and limitations of extrapolating findings from animal studies to clinical settings. Furthermore, this paper recommends adopting the Grading of Recommendations Assessment, Development and Evaluation (GRADE) methodology to comprehensively grade the quality of evidence. Through a modular analysis of the entire reporting process, this paper aims to provide researchers in the field with a clear and practical guide, thereby promoting the standardized development of systematic reviews and meta-analysis of animal experiments and enhancing their application value in scientific decision-making and translational medicine.

Objective To investigate the expression of α7 nicotinic acetylcholine receptor (α7nAChR) in thymic T follicular helper cells (TFH) and its significance in patients with myasthenia gravis (MG). Methods Fifteen MG patients who underwent surgical treatment at the Myasthenia Gravis Comprehensive Diagnosis and Treatment Center of Henan Provincial People’s Hospital from June 2022 to June 2023 were selected as a MG group, including 7 males and 8 females, aged 12-30 years. Twelve patients who underwent partial thymectomy to optimize surgical field exposure during cardiac surgery at Fuwai Central China Cardiovascular Hospital from June 2022 to June 2023 were selected as a control group, including 5 males and 7 females aged 20-35 years. Thymus single cell suspension was obtained by grinding the thymus tissue, and flow cytometry was used to detect the expression of α7nAChR in TFH cells. The thymus cell suspension was purified using density gradient centrifugation, followed by immunomagnetic bead separation to obtain CD4+T cells. CXCR5 antibody and coupled magnetic beads were added to isolate TFH cells. Real-time fluorescent polymerase chain reaction and Western blotting were performed to further investigate the expression of α7nAChR in TFH cells. Results Compared with the control group, the proportion of thymic TFH cells in the MG group was significantly increased (P<0.05), along with significantly decreased mRNA and protein expression levels of α7nAChR within these cells (P<0.01). Conclusion The findings suggest that there is a reduced expression of α7nAChR within thymic TFH cells in MG patients, leading to weakened immunosuppressive function which may indirectly contribute to disease onset and progression.

To investigate the role of the DPP4-nestin axis in liver fibrosis induced by alveolar cyst infection,a murine model was established using C57BL/6 mice via hepatic portal vein injection.Liver histopathological changes were assessed using HE staining,while immunohistochemistry and immunofluorescence were employed to evaluate the expression levels of nestin and DPP4 in infected mouse livers.In vitro,J S1 cell line was stimulated with recombinant DPP4 protein to es-tablish a cellular model,and qPCR,Western blot,and shRNA lentivirus interference techniques were utilized to examine the involvement of the DPP4-nestin axis in hepatic stellate cell activation.The findings demonstrated that compared to the Sham group,liver tissue structure disruption and collagen deposition were evident along with significantly increased expressions of nestin and DPP4(P＜0.050 0),which colocalized with nesin and α-SMA.Furthermore,stimulation with recombi-nant DPP4 protein significantly enhanced JS1 cell activation(P＜0.050 0)as well as upregulated nestin expression(P＜0.050 0)when compared to control group cells.Notably,shRNA lentivirus-mediated inhibition of nestin expression effectively suppressed the activating effects exerted by re-combinant DPP4 protein on JS1 cells(P＜0.050 0).Collectively,these results highlight the crucial regulatory role played by the DPP4-nestin axis in hepatic stellate cell activation triggered by alveo-lar infection;thus,targeting this axis may represent a novel therapeutic strategy for treating alveo-lar infection-induced liver fibrosis.

Chronic pancreatitis is a chronic inflammatory disease characterized by progre-ssive fibrosis of pancreatic tissue. Its pathological features primarily include parenchymal fibrosis, intraductal stone formation or calcification deposits, as well as segmental stenosis and dilation of the pancreatic duct. Prolonged chronic inflammatory stimulation not only leads to progressive pancreatic dysfunction but may also trigger the formation of pancreatic pseudocysts and even malignant transformation. In the comprehensive treatment of chronic pancreatitis, the core clinical goals are the removal of pancreatic duct stones, restoration of unobstructed pancreatic duct drainage, and preservation of residual pancreatic function. Traditional treatment strategies have been based on the principle of progressive intervention and early surgical management. In recent years, with advancements in extracorporeal shock wave lithotripsy, the application of new techniques such as endoscopic retrograde cholangiopancreatography combined with laser lithotripsy under direct cholan-gioscopic visualization, and improvements in pancreas-preserving surgical approaches, the debate over the superiority of progressive intervention versus early surgical treatment has intensified. Against this backdrop, the treatment mode of Xi′an Jiaotong University Pancreatic Disease Center (hereinafter referred to as "Western Pancreas") has emerged, emphasizing a personalized, multimodal treatment strategy based on different types of pancreatic duct stones. The treatment mode of "Western Pancreas" integrates lithotripsy, endoscopic treatment, and surgical interventions to optimize patient outcomes. By conducting a comprehensive analysis of domestic and international pancreatic duct stone classi-fication systems and drawing from our team′s clinical experience in managing over a thousand cases of chronic pancreatitis, the authors have further refined and proposed a classification system for pancreatic duct stones under the treatment mode of "Western Pancreas". This refinement aims to enhance the overall diagnostic and therapeutic standards for chronic pancreatitis.

Objective To investigate the effect of butorphanol on lipopolysaccharide-induced chondrocyte injury by regulating the stromal cell-derived factor-1α(SDF-1α)/C-X-C chemokine receptor 4(CXCR4)pathway.Methods Human C28/12 chondrocytes were cultured in vitro and assigned to the following groups:control(normal culture),model(100 μmol/L lipopolysaccharide),model+low-dose butorphanol(100 μmol/L lipopolysaccharide+1μmol/L butorphanol),model+medium-dose butorphanol(100 μmol/L lipopolysaccharide+2 μmol/L butorphanol),model+high-dose butorphanol(100 μmol/L lipopolysaccharide+4 μmol/L butorphanol),and model+high-dose butorphanol+NUCC-390(100 μmol/L lipopolysaccharide+4 μmol/L butorphanol+500 nmol/L CXCR4 agonist NUCC-390).Cell viability,interleukin(IL)-6 and tumor necrosis factor-α(TNF-α)levels,apoptosis,and SDF-1α/CXCR4 pathway-related proteins were evaluated by MTT assay,enzyme-linked immunosorbent assay,flow cytometry,and Western blot,respectively.Results Chondrocyte survival rate and Bcl-2 protein expression were decreased while TNF-α,IL-6,apoptosis rate,Bax,Cleaved caspase-3,SDF-1α,and CXCR4 proteins were increased in the model group compared with the control group(P＜0.05).The above indicators were improved in the model+low-,medium-,and high-dose butorphanol groups compared with the model group,while the result for the model+high-dose butorphanol+NUCC-390 group were opposite to those of the model+high-dose butorphanol group.Conclusions Butorphanol may improve lipopolysaccharide-induced chondrocyte injury induced by inhibiting the SDF-1α/CXCR4 signaling pathway.

Swine enteric coronaviruses(SeCoV),such as porcine epidemic diarrhea virus(PEDV),transmissible gastroenteritis virus(TGEV),and porcine delta coronavirus(PDCoV),cause severe diarrhea in piglets,resulting in substantial losses in pig farming.In this study we establish a triple fluorescence reverse transcription-quantitative PCR(RT-qPCR)method for the simultaneous de-tection of PEDV,TGEV,and PDCoV.The specific primers and probes for each target virus were designed based on conserved sequences from the PEDV M gene,the TGEV ORF 1b gene,and the PDCoV ORF 1b gene respectively.Following the optimization of parameters and conditions,a triple RT-qPCR method was successfully established to simultaneously detect PEDV,TGEV,and PD-CoV.The developed assay exhibits strong specificity for these three pathogens without any cross-reaction with other common porcine viruses like CSFV,PCV2,PoRVA,PRV,and PRRSV.The de-tection limit of linear templates for pTOPO-PEDV 128,pTOPO-TGEV 116,and pTOPO-PDCoV 125 recombinant plasmids were 16.835,17.610 and 17.020 copies/μL,respectively.The intra group and inter group coefficients of variation were less than 5％,with no significant differences observed(P＞0.05).Moreover,the detection consistency rate of the developed RT-qPCR was compared with standard method and showed 100％agreement.Out of 35 small intestine tissue samples,17 tested positive for PEDV,resulting in a positive rate of 48.57％(17/35).The tests for TGEV and PDCoV yielded negative results,and no mixed infections were detected.Based on the above results,the tri-ple RT-qPCR method established is specific,sensitive,stable,and rapid,and can be used for clinical detection and differential diagnosis of PEDV,TGEV,and PDCoV simultaneously,providing a method for the detection and epidemiological investigation of porcine diarrhea coronaviruses.

Chronic pancreatitis is a chronic inflammatory disease characterized by progre-ssive fibrosis of pancreatic tissue. Its pathological features primarily include parenchymal fibrosis, intraductal stone formation or calcification deposits, as well as segmental stenosis and dilation of the pancreatic duct. Prolonged chronic inflammatory stimulation not only leads to progressive pancreatic dysfunction but may also trigger the formation of pancreatic pseudocysts and even malignant transformation. In the comprehensive treatment of chronic pancreatitis, the core clinical goals are the removal of pancreatic duct stones, restoration of unobstructed pancreatic duct drainage, and preservation of residual pancreatic function. Traditional treatment strategies have been based on the principle of progressive intervention and early surgical management. In recent years, with advancements in extracorporeal shock wave lithotripsy, the application of new techniques such as endoscopic retrograde cholangiopancreatography combined with laser lithotripsy under direct cholan-gioscopic visualization, and improvements in pancreas-preserving surgical approaches, the debate over the superiority of progressive intervention versus early surgical treatment has intensified. Against this backdrop, the treatment mode of Xi′an Jiaotong University Pancreatic Disease Center (hereinafter referred to as "Western Pancreas") has emerged, emphasizing a personalized, multimodal treatment strategy based on different types of pancreatic duct stones. The treatment mode of "Western Pancreas" integrates lithotripsy, endoscopic treatment, and surgical interventions to optimize patient outcomes. By conducting a comprehensive analysis of domestic and international pancreatic duct stone classi-fication systems and drawing from our team′s clinical experience in managing over a thousand cases of chronic pancreatitis, the authors have further refined and proposed a classification system for pancreatic duct stones under the treatment mode of "Western Pancreas". This refinement aims to enhance the overall diagnostic and therapeutic standards for chronic pancreatitis.

Objective To investigate the effect of butorphanol on lipopolysaccharide-induced chondrocyte injury by regulating the stromal cell-derived factor-1α(SDF-1α)/C-X-C chemokine receptor 4(CXCR4)pathway.Methods Human C28/12 chondrocytes were cultured in vitro and assigned to the following groups:control(normal culture),model(100 μmol/L lipopolysaccharide),model+low-dose butorphanol(100 μmol/L lipopolysaccharide+1μmol/L butorphanol),model+medium-dose butorphanol(100 μmol/L lipopolysaccharide+2 μmol/L butorphanol),model+high-dose butorphanol(100 μmol/L lipopolysaccharide+4 μmol/L butorphanol),and model+high-dose butorphanol+NUCC-390(100 μmol/L lipopolysaccharide+4 μmol/L butorphanol+500 nmol/L CXCR4 agonist NUCC-390).Cell viability,interleukin(IL)-6 and tumor necrosis factor-α(TNF-α)levels,apoptosis,and SDF-1α/CXCR4 pathway-related proteins were evaluated by MTT assay,enzyme-linked immunosorbent assay,flow cytometry,and Western blot,respectively.Results Chondrocyte survival rate and Bcl-2 protein expression were decreased while TNF-α,IL-6,apoptosis rate,Bax,Cleaved caspase-3,SDF-1α,and CXCR4 proteins were increased in the model group compared with the control group(P＜0.05).The above indicators were improved in the model+low-,medium-,and high-dose butorphanol groups compared with the model group,while the result for the model+high-dose butorphanol+NUCC-390 group were opposite to those of the model+high-dose butorphanol group.Conclusions Butorphanol may improve lipopolysaccharide-induced chondrocyte injury induced by inhibiting the SDF-1α/CXCR4 signaling pathway.