pAPN is a zinc-dependent metalloprotease,mediating the fusion between virus and host cell,and playing a role as the receptor of coronavirus.To explore the effect of pAPN on PEDV rep-lication,the full-length pAPN gene was amplified from the porcine small intestinal by PCR,and was cloned into the lentiviral vector via the homologous site digested with BamH Ⅰ and Not Ⅰ to obtain the recombinant lentiviral vector PLVX-pAPN-mCMV-ZsGreen1-puro.The recombinant lentiviral vector and helper plasmids pLP1,pLP2,pLP-VSVG were co-transfected into 293T cells for lentiviral packaging.Vero cells were infected with the packaged lentivirus and the pAPN gene overexpressing cells were screened by puromycin.The stable expression of Vero-pAPN monoclonal cell line was screened by a limited dilution method,and the effect of the cell line on the replication of PEDV was determined by qPCR for N mRNA transcription level,Western blot for N protein level,and TCID50.The results showed that the packaged lentivirus could infect Vero cells,and the monoclonal cell line Vero-pAPN(2C5)could stably expressed pAPN.The Vero-pAPN cell line can promote the replication of PEDV,the N gene mRNA transcription level was significantly different at 12-48 h(P＜0.05),the N protein expression level increased,and the TCID50 was significantly different at 24 and 48 h(P＜0.05).In conclusion,the Vero-pAPN cell line was constructed in this study and it can significantly promote the replication of PEDV,which provides a candidate cell line for PEDV vaccine production and isolation.

Danggui Buxue decoction(DBD)is a classic prescription with immunomodulatory and hematopoietic effects.Previous studies have shown the DBD has potential to be used as an oral im-mune booster.However,its immunomodulatory effects and mechanism of action have not been thoroughly studied,especially the protective mechanism of immunomodulatory regulation in the state of immunosuppressive is still unclear.The aim of this study was to explore the protective mechanism of DBD in the immunosuppressive state using network pharmacology combined with animal experiments verification.The active components,core targets and signaling pathways of DBD in treating immunosuppression were obtained using network pharmacology tools.On this ba-sis,the active components of DBD were identified using HPLC-MS,and in vivo studies were con-ducted at the same time.The key active components of DBD obtained using network pharmacology included quercetin,kaempferol and formononetin.The core targets included TP53,RELA,TNF,AKT1,and IL-6.KEGG pathway enrichment analysis showed that phosphoinositide 3-kinase(PI3K)-protein kinase B(AKT)may play an important role in the treatment of immunosuppres-sive diseases using DBD.Molecular docking confirmed that each core target had good binding activ-ity with its corresponding compounds.Animal experiments showed that after DBD intervention,the mRNA gene and protein expression of RELA,TNF,and IL-6 in the serum was significantly down-regulated.The mRNA expression of PI3K and AKT in the ileum and PI3K protein expression were also downregulated.In conclusion,DBD exerts its role in treating immunosuppressive diseases by regulating the PI3K-AKT signaling pathway.

OBJECTIVE To study the constituents from the dried stems and leaves of Illicium dunnianum Tutcher.METHODS The compounds were isolated and purified by column chromatography of AB-8 macroporous resin,silica gel,HW-40C,ODS,Sephadex LH-20,and semi-preparative RP-HPLC.Their structures were elucidated by physicochemical properties,spectral analyses and ECD.RESULTS The 70%ethanol extract of Illicium dunnianum was subjected to AB-8 macroporous adsorption resin CC to yield 30%ethanol fraction.Five compounds were obtained and characterized as anisole glycol-7-O-β-L-funanarabifuranosyl-(1→6)-β-D-glucopyranoside(1),4-O-β-D-glucopyranosyloxy-benzaldehyde(2),benzyl-O-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside(3),β-D-glucopyranoside benzoate(4)and sachalinoside B(5),respectively.CONCLUSION Compound 1 is a new phenolic glycoside,2-5 are identified from Illicium dunnianum for the first time.

Objective:To investigate the dosimetric advantages of deep inspiration breath-hold (DIBH) technique in postoperative radiotherapy for left-sided breast cancer.Methods:A prospective study was conducted on patients requiring adjuvant radiotherapy after left-sided breast cancer surgery at Jiangsu Cancer Hospital from January 2018 to May 2023. CT simulation images were acquired under both free breathing (FB) and DIBH respiratory modes. Planning target volumes (PTV) and organs at risk (OAR) were delineated, and dosimetric parameters were compared between the two respiratory modes. Additionally, patients were grouped into subgroups [internal mammary lymph node irradiation (IMNI) vs. non-IMNI, breast-conserving surgery (BCS) followed by radiotherapy vs. modified radical mastectomy (MRM) followed by radiotherapy], and dosimetric differences among subgroups for both breathing modes were compared. The Velocity system was used to measure the minimum distances from the heart and left anterior descending coronary artery (LAD) to the PTV surface on CT images. These distances were defined as the heart-to-PTV and LAD-to-PTV distances. Pearson correlation analysis was performed to assess the relationships between heart D max and LAD D max, heart-to-PTV distance and heart D mean, and LAD-to-PTV distance and LAD D max under both respiratory modes. Results:A total of 132 patients were included. Compared to the FB, DIBH showed no significant difference in target dose distribution, but significantly reduced dose to OAR. Specifically, the heart D mean and D max decreased by 1.8 Gy and 8.1 Gy, respectively, and the LAD D max decreased by 7.9 Gy, and the affected lung V 5 Gy and V 20 Gy were reduced by 6.4% and 2.5%, respectively (all P<0.05). All subgroups benefited from DIBH, with greater decrease of dose to OAR in the IMNI subgroup (compared with the non-IMNI subgroup) and the subgroup of MRM followed by radiotherapy (compared with the BCS followed by radiotherapy group). Under both FB and DIBH modes, heart D max and LAD D max showed linear correlations ( r=0.62 and 0.84, respectively; both P<0.001), heart-to-PTV distance correlated with heart D mean ( r=-0.61 and -0.67, respectively; both P<0.001), and LAD-to-PTV distance correlated with LAD D max ( r=-0.58 and -0.63, respectively; both P<0.001). Conclusions:The DIBH technique can significantly reduce dose to the heart, LAD, and lungs in patients undergoing postoperative radiotherapy for left-sided breast cancer without compromising target dose. Patients receiving IMNI after left-sided breast cancer surgery benefit more from the DIBH technique.

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.

Neoadjuvant therapy for hepatocellular carcinoma is the frontier and hot topic in the current field of liver cancer research.The fundamental purpose is to reduce the risk of postoperative recurrence through standardized preoperative treatment methods.From the attempts of Transcatheter Arterial Chemoembolization monotherapy for neoadjuvant therapy for hepatocellular carcinoma to systematic treatment represented by"targeted combined with immunotherapy",the latter has become the most promising neoadjuvant strategy due to its high objective response rate and potential to induce pathological complete remission.However,the field still faces challenges such as lack of evidence of overall survival benefit in Phase Ⅲ randomized controlled trials,treatment-related adverse reactions that may lead to delay in surgery,optimal population screening,and timing of surgery.This article aims to briefly discuss the current research status of the application of neoadjuvant therapy in resectable hepatocellular carcinoma,explore relevant diagnosis and treatment concepts,and further understand neoadjuvant therapy.

Objective:To screen broadly neutralizing antibodies in human immunodeficiency virus-1（HIV-1）chronically infected individuals and characterize their molecular features and to provide new strategies for rational vaccine development and antibody-based therapeutics.Methods:A total of 34 treatment-na?ve individuals with chronic HIV-1 infection were enrolled. Plasma antibody binding levels were measured against two HIV-1 envelope proteins. Single antigen-specific memory B cells were sorted from high-binding samples，and antibody variable region genes were amplified by PCR for paired expression. The monoclonal antibodies were evaluated for neutralizing activity using pseudovirus assays，and their structural features were analyzed by integrating AlphaFold 3 prediction with Discovery Studio molecular docking.Results:Plasma samples showed strong binding to DU422-GP140 and BG505-GP140. Eight monoclonal antibodies were isolated from two donors. Among them，antibodies 0919-A4，0919-A9 and 0808-A2 could cross-react with GP140 from HIV-1 subtypes AE，BC and B. The monoclonal antibody 0919-A9 demonstrated potent neutralizing activity against SF162（Tier 1）and CH181（Tier 2）pseudoviruses，with somatic hypermutation rates of 13.27%（heavy chain）and 15.58%（light chain）. Structural modeling revealed its specific targeting of the V1V2 region on GP120.Conclusion:The isolated antibody 0919-A9 effectively neutralizes Tier 2 pseudoviruses. Its high somatic mutation frequency and V1V2-targeting property underlie its neutralizing activity，providing both a promising candidate and mechanistic insights for HIV vaccine development and antibody-based therapeutic strategies.

Objective:To evaluate the safety and effectiveness of fuzuloparib for the treatment of ovarian epithelial cancer patients in the real world setting.Methods:A retrospective analysis was conducted on the baseline data of 4 620 ovarian cancer patients who had received fuzuloparib monotherapy or combination therapy. Another 224 ovarian cancer patients who were willing to receive fuzuloparib monotherapy or combination therapy were prospectively enrolled, and their baseline characteristics, drug effectiveness, and safety data were analyzed.Results:(1) Among the 4 620 patients in the retrospective cohort, the median age of patients was 60 years; tumor types: 89.8% (4 149/4 620) had ovarian cancer. Among patients with clearly documented information, the vast majority had a histological type of serous carcinoma (82.9%, 3 770/4 546) and International Federation of Gynecology and Obstetrics (FIGO) staging of Ⅲ-Ⅳ (90.9%, 1 537/1 691). (2) Among the 224 patients in the prospective cohort, the median age of patients was 57 years; tumor types: 83.9% (188/224) had ovarian cancer. Among patients with clearly documented records, the predominant pathologic type was serous carcinoma (91.9%, 193/210), and FIGO stage was Ⅲ-Ⅳ in 79.9% (139/174). (3) Among the 224 prospective patients: 84 patients received first-line fluzoparib maintenance therapy, 92 patients received fluzoparib maintenance therapy after platinum-sensitive recurrence, 23 patients received direct fluzoparib treatment after platinum-sensitive recurrence, 19 patients received direct fluzoparib treatment after platinum-resistant recurrence. The median follow-up durations were 8.5, 8.7, 7.9, and 6.7 months, respectively. The median durations of fluzoparib treatment were 6.7, 4.8, 3.1, and 1.9 months, respectively. The median progression-free survival (PFS) times were not reached during follow-up, 12.6 months, not reached during follow-up, and 4.8 months, respectively. The 1-year PFS rates were 84.1%, 55.0%, 69.8%, and 45.5%, respectively. The remaining 6 patients received other fluzoparib regimens. (4) Among the 224 patients in the prospective dataset, 205 had safety data recorded. Of these, 127 patients (62.0%, 127/205) experienced treatment-related adverse events, with common events including anemia (24.4%, 50/205), thrombocytopenia (21.0%, 43/205), and leukopenia (19.5%, 40/205). Among the 205 patients, 43 (21.0%, 43/205) experienced grade 3 or higher treatment-related adverse events, with common events including anemia (8.3%, 17/205) and thrombocytopenia (8.3%, 17/205).Conclusions:The effectiveness of fuzuloparib in clinical application is generally consistent with other drugs in the same class, with good safety. This study provids new clinical evidence for the treatment of ovarian cancer with fuzuloparib.

Objective:To compare the efficacy of new nasopharyngeal airway and laryngeal mask airway for airway management in the patients undergoing non-intubated video-assisted thoracoscopic surgery (NIVATS).Methods:In this randomised, controlled, non-inferiority trial, 60 American Society of Anesthesiologists Physical Status classification I or Ⅱ patients of both sexes, aged 18-79 yr, scheduled for elective NIVATS from December 2021 to December 2023 at Jining No.1 People′s Hospital, were divided into 2 groups ( n=30 each) using a computer-generated random code in a 1∶1 ratio: new type nasopharyngeal airway group (group N) and laryngeal mask airway group (group L). After anesthesia induction, a new nasopharyngeal airway was inserted in group N, and a laryngeal mask airway was inserted in group L. Spontaneous ventilation was maintained during the NIVATS. Ultrasound-guided serratus anterior plane block was performed on the affected side before anesthesia induction. Anesthesia was maintained with propofol and remifentanil. The primary outcome measure was the rate of intraoperative airway intervention, the airway interventions included repositioning of the airway tools, manual assisted ventilation, jaw-thrust maneuver, and conversion to endotracheal intubation. The secondary outcome measures included the first-attempt success rate of airway device placement, time for establishing a patent airway, the minimum value of SpO 2, the maximum value of P ETCO 2, and incidence of complications such as postoperative sore throat. Results:The rate of intraoperative airway intervention was 27% in group L and 47% in group N ( χ2=2.58, P=0.108). The difference in the rate of intraoperative airway intervention between the two groups was 0.20 (95% confidence interval 0.15-0.25), with a 95% confidence interval upper limit higher than the non-inferiority boundary (10%), indicating that this non-inferiority hypothesis was not established. In comparison to group L, the rate of intraoperative jaw-thrust maneuver intervention was significantly increased, the time to establish a patent airway was shortened, and the incidence of postoperative sore throat was decreased in group N ( P<0.05). Conclusions:Compared with the laryngeal mask airway, the new nasopharyngeal airway can reduce the development of postoperative throat pain, however, it is less effective in maintaining a patent airway. It requires careful consideration of risks and benefits when used for NIVATS.

Aim To investigate the regulatory effects of coenzyme Q10(CoQ10)on blood lipid level and intesti-nal flora abundance in atherosclerotic(As)rats.Methods 24 rats were randomly divided into control group,As group and CoQ10 intervention group.Rats in the As group and CoQ10 intervention group were fed with high-fat chow for 2 weeks,combined with abdominal aortic balloon injury to replicate the As model.CoQ10 was administered by gavage start-ing on the next day of modeling,once daily for 4 weeks.Aortic Movat's staining and lipid levels were used to verify the effect of CoQ10 intervention in As,and the abundance of intestinal flora in intestinal contents was analyzed using met-agenomics.Results Compared with the control group,rat aortic tissues in the As group showed endothelial damage,structural disorganization of the internal elastic plate and inflammatory infiltration,serum triglyceride(TG),total cholester-ol(TC)and low density lipoprotein cholesterol(LDLC)levels were increased,and high density lipoprotein cholesterol(HDLC)levels were decreased.Compared with the As group,the structure of the endothelial cells of the aorta and the structure of the endothelial cells,the internal elastic plates and the smooth muscle cell morphology were relatively regular,serum TC,TG and LDLC levels were decreased,and HDLC levels were increased in the CoQ10 intervention group.Com-pared with the control group,the intestinal bacterial biodiversity in the As group was reduced.At the phylum level,the abundance of the Firmicutes and the Bacteroidetes were down-regulated,whereas that of Proteobacteria was up-regulated.At the genus level,the relative abundance of Lactobacillus,Bacteroides,Akkermansia,Limosilactobacillus,Parabacteroides and Ligilactobacillus was down-regulated,and the relative abundance of Muribaculum was up-regulated.Compared with the As group,CoQ10 intervention restored the biodiversity of the intestinal microbiota in As rats and increased the relative abundance of Firmicutes,Bacteroidetes,Lactobacillus,Bacteroides,Akkermansia,Limosilactobacillus,Parabacteroides and Ligilactobacillus,while reducing the relative abundance of Proteobacteria and Muribaculum(all P＜0.05).Conclusion CoQ10 can regulate blood lipid levels in As rats,upregulate the abundance of beneficial microbiota,downregulate the abun-dance of harmful microbiota,and modulate the diversity of the gut microbiota.