ObjectiveTo investigate the effects of forkhead box protein O4 （FOXO4） expression on gastric precancerous lesions （GPL）， and to clarify its mechanism in mediating the therapeutic effect of Celastrus orbiculatus extract （COE） on GPL by regulating aerobic glycolysis. MethodsReferring to the previously established combined modeling protocol in our research group， a rat model of gastric precancerous lesions （GPL） was constructed through the following procedures： rats were given free access to 170 μg/mL N-methyl-N′-nitro-N-nitrosoguanidine （MNNG） solution for ad libitum drinking， fed with a diet supplemented with 0.03% ranitidine hydrochloride， and treated with a cycling regimen of “2-day feeding followed by 1-day fasting”. Specifically， on the afternoon of each fasting day， the rats received intragastric administration of 2% sodium salicylate at a dose of 10 mL/kg body weight. FOXO4-silenced and overexpression model rats were constructed by tail vein injection of plasmids. The rats were randomly divided into control， model， COE， overexpression negative control （OE-NC）， FOXO4 overexpression （OE-FOXO4）， OE-FOXO4+COE， silencing negative control （shNC）， FOXO4 silencing （shFOXO4） and shFOXO4+COE groups. Gastric mucosal histopathological changes were observed in each group. Lactic acid content in gastric mucosal tissues was detected by colorimetry. The expression levels of FOXO4， HK2， PKM2， LDHA and GLUT1 were evaluated by immunohistochemistry （IHC）， and their mRNA levels were determined by RT-PCR.Results Compared with the control group， the COE， OE-FOXO4 and OE-FOXO4+COE groups exhibited significantly improved gastric mucosal lesions， reduced lactic acid levels， weakened expression of aerobic glycolysis-related proteins （PKM2， HK2， LDHA， GLUT1）， and enhanced FOXO4 expression. The OE-FOXO4+COE group showed the lowest lactic acid level and more pronounced changes in related protein expression compared with the COE and OE-FOXO4 groups. In contrast， the shFOXO4 and shFOXO4+COE groups displayed increased lactic acid levels， enhanced expression of aerobic glycolysis-related proteins， and reduced FOXO4 expression compared with the model group.ConclusionFOXO4 expression is involved in the inhibitory effect of COE on GPL， possibly by regulating the aerobic glycolysis process.

ObjectiveTo identify the direct target proteins for the anti-obesity effect of resveratrol （Res） via chemical proteomic analysis and systematically explore the molecular mechanisms by which Res inhibits lipid accumulation in adipocytes from mitochondrial function regulation， adipocyte browning， and inflammation inhibition， thus providing new targets and experimental evidence for the prevention and treatment of obesity and related metabolic diseases. MethodsThe cell counting kit-8 （CCK-8） assay was adopted to detect the safe and effective doses （7.5， 15， 30 μmol·L^-1） of Res for treating 3T3-L1 cells based on the cell survival rate. After being induced to differentiate into mature adipocytes， the cells were assigned into the blank ［treated with dimethyl sulfoxide （DMSO）］ group and the Res （7.5， 15， 30 μmol·L^-1） groups and treated for 48 h. Oil red O staining and total triglyceride （TG） content were assessed to measure lipid accumulation in adipocytes. Target protein identification of Res was conducted through IAA-P-based chemical proteomics combined with high-resolution mass spectrometry. Target proteins of Res were validated by cellular thermal shift assay （CETSA） combined with Western blot analysis. Furthermore， the expression of browning markers peroxisome proliferator-activated receptor γ coactivator-1α （PGC-1α） and mitochondrial uncoupling protein 1 （UCP1）， as well as the target proteins NADH-ubiquinone oxidoreductase core subunit S8 （NDUFS8） and high-mobility group protein B1 （HMGB1） were determined by Western blot. The ATP assay kit was used to measure the ATP concentration. Mitochondrial biogenesis was characterized with MitoTracker Green， on the basis of which the regulatory effect of Res on mitochondrial function was analyzed. The level of the inflammatory cytokine tumor necrosis factor-α （TNF-α） was measured by enzyme-linked immunosorbent assay （ELISA）， on the basis of which the anti-inflammatory effect of Res on adipocytes was evaluated. ResultsCompared with the control group， Res demonstrated safety and efficacy within the range of 7.5-30 μmol·L^-1， without significantly affecting cell viability. Compared with the control group， different concentrations of Res reduced lipid accumulation in 3T3-L1 adipocytes （P<0.01）， promoted the expression of NDUFS8 （P<0.01）， increased the ATP concentration and the protein levels of UCP1 and PGC-1α， elevated the level of TNF-α （P<0.05， P<0.01）. ConclusionNDUFS8 and HMGB1were identified as the target proteins of Res in mitigating obesity. Res promotes mitochondrial biogenesis to induce the browning of 3T3-L1 adipocytes via targeting NDUFS8 and it inhibits the TNF-α-mediated inflammatory signaling via targeting HMGB1， thus inhibiting lipid accumulation and ameliorating obesity.

Objective To explore the relationship between PANoptosis and hepatic ischemia-reperfusion injury (HIRI), and to screen the key genes of PANoptosis in HIRI. Methods PANoptosis-related differentially expressed genes (PDG) were obtained through the Gene Expression Omnibus database and GeneCards database. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Set Enrichment Analysis (GSEA) were used to explore the biological pathways related to PDG. A protein-protein interaction network was constructed. Key genes were selected, and their diagnostic value was assessed and validated in the HIRI mice. Immune cell infiltration analysis was performed based on the cell-type identification by estimating relative subsets of RNA transcripts. Results A total of 16 PDG were identified. GO analysis showed that PDG were closely related to cellular metabolism. KEGG analysis indicated that PDG were mainly enriched in cellular death pathways such as apoptosis and immune-related signaling pathways such as the tumor necrosis factor signaling pathway. GSEA results showed that key genes were mainly enriched in immune-related signaling pathways such as the mitogen-activated protein kinase (MAPK) signaling pathway. Two key genes, DFFB and TNFSF10, were identified with high accuracy in diagnosing HIRI, with areas under the curve of 0.964 and 1.000, respectively. Immune infiltration analysis showed that the control group had more infiltration of resting natural killer cells, M2 macrophages, etc., while the HIRI group had more infiltration of M0 macrophages, neutrophils, and naive B cells. Real-time quantitative polymerase chain reaction results showed that compared with the Sham group, the relative expression of DFFB messenger RNA in liver tissue of HIRI group mice increased, and the relative expression of TNFSF10 messenger RNA decreased. Cibersort analysis showed that the infiltration abundance of naive B cells was positively correlated with DFFB expression (r=0.70, P=0.035), and the infiltration abundance of M2 macrophages was positively correlated with TNFSF10 expression (r=0.68, P=0.045). Conclusions PANoptosis-related genes DFFB and TNFSF10 may be potential biomarkers and therapeutic targets for HIRI.

Objective : To observe the effect of Celastrus orbiculatusextract(COE) on gastric precancerous lesions(GPL) and to explore its role in the Notch-1 signaling pathway. Methods : GPL rat models were established using a composite model replication method, and the rats were randomly divided into a control group, a model group, and COE low, medium and high dose groups [COE at 12.5, 25, and 50 mg/(kg·d)]. After 4 weeks of intervention, gastric tissue was collected, and immunohistochemistry(IHC) was performed to detect the expression of mucins(MUC2, MUC5AC, and MUC6), Leucine-rich repeat-containing G-protein coupled receptor 5(Lgr5), Proliferating Cell Nuclear Antigen(Ki67), and Notch-1. Polymerase chain reaction(PCR) was used to determine the mRNA levels of the aforementioned mucins. Human gastric epithelial cells(GES-1) were induced with N-Methyl-N′-nitro-N-nitrosoguanidine(MNNG) to establish a GPL cell model. The cells were randomly divided into control, model, and COE low, medium, and high concentration groups(COE at 5, 10, and 20 μg/ml). After 24 hours of corresponding interventions, changes in cell morphology were observed under an inverted microscope. Western blot was used to measure the expression of Notch-1 and Lgr5, and immunofluorescence(IF) was employed to detect Notch-1 expression. Results : Compared to the control group, the expression of MUC2, Lgr5, Notch-1, and Ki67 in the gastric tissue of the model group rats significantly increased(P<0.000 1), while the expression of MUC5AC and MUC6 decreased(P<0.000 1). In comparison to the model group, the expressions of MUC2, Lgr5, Notch-1, and Ki67 were significantly reduced in the COE groups(P<0.01), while the expression of MUC5AC and MUC6 significantly increased(P<0.01). In the GES-1 model group, the cells exhibited irregular morphology, loose intercellular connections, and disorganized arrangement compared to the control group. In contrast, the cells in the COE groups displayed a more regular morphology and a more organized arrangement than those in the model group. Additionally, compared to the control group, the expression of Lgr5 and Notch-1 in the model group were significantly elevated(P<0.000 1), whereas after COE treatment, their expressions were markedly reduced(P<0.001). Conclusion COE can alleviate GPL, and its mechanism may be associated with the downregulation of the Notch-1 signaling pathway, which improves gastric mucosal mucin barrier function and inhibits the abnormal proliferation of gastric mucosal stem cells.

Objective:To evaluate the efficacy and safety of remedial pedicled rectus abdominis myocutaneous flap transplantation for breast reconstruction.Methods:This retrospective study included 23 female breast cancer patients who underwent autologous breast reconstruction with a remedial pedicled rectus abdominis myocutaneous flap at Hunan Cancer Hospital from June 2022 to June 2023. The patients' ages ranged from 24 to 56 years, with a mean age of (43.2±1.3) years. All patients had previously undergone mastectomy and tumor resection before receiving the remedial pedicled rectus abdominis myocutaneous flap reconstruction. Follow-up was conducted through outpatient visits and telephone interviews, with a median follow-up period of 10 (6-15) months. Observation indicators included survival rate of flaps, patient satisfaction rate and aesthetic outcomes. Adverse reactions including but not limited to flap contracture and deformation, during and following the surgical procedure were also recorded.Results:The remedial pedicled rectus abdominis myocutaneous flap technique was utilized for autologous breast reconstruction in 23 patients. Specifically, this approach was chosen due to two primary reasons: the perforator distance from the vascular pedicle being excessively long in 18 cases, and suboptimal characteristics of the main pedicle in 5 cases. Two types of flap configurations were employed. For 15 patients, a unilateral remedial pedicled rectus abdominis myocutaneous flap was used, with an average flap size of (21.5±0.8) cm in length and (11.2±1.3) cm in width. All flaps demonstrated adequate blood supply without the need for additional vascular anastomosis. For the remaining 8 patients, a combination of a unilateral remedial pedicled rectus abdominis myocutaneous flap and a contralateral free lower abdominal flap was utilized, resulting in an average flap size of (29.3±0.7) cm in length and (12.0±1.8) cm in width. In these cases, the inferior epigastric vessels of the free lower abdominal flap were successfully anastomosed to the internal thoracic vessels of the recipient site, ensuring all flaps survived postoperatively. The survival rate of flaps in 23 patients was 100% (23/23), and the patient satisfaction rate was 100% (23/23). Postoperative evaluation revealed that the reconstructed breasts exhibited satisfactory shape and texture, with no instances of flap contracture or deformation. The reconstructed breasts were largely symmetrical with the contralateral side, leaving only linear scars at the donor and recipient sites. Importantly, there was no significant impact on abdominal wall function, and no adverse reactions such as flap contracture or impaired abdominal wall function were observed in any of the patients.Conclusion:When the characteristics of the perforating vessels in the lower abdomen are suboptimal, the use of a remedial pedicled rectus abdominis myocutaneous flap technique for breast reconstruction can ensure satisfactory outcomes and significantly enhance surgical safety.

Objective To investigate the distribution and antimicrobial resistance profiles of clinically isolated Haemophilus influenzae and Moraxella catarrhalis in hospitals across China from 2015 to 2021,and provide evidence for rational use of antimicrobial agents.Methods Data of H.influenzae and M.catarrhalis strains isolated from 2015 to 2021 in CHINET program were collected for analysis,and antimicrobial susceptibility testing was performed by disc diffusion method or automated systems according to the uniform protocol of CHINET.The results were interpreted according to the CLSI breakpoints in 2022.Beta-lactamases was detected by using nitrocefin disk.Results From 2015 to 2021,a total of 43 642 strains of Haemophilus species were isolated,accounting for 2.91％of the total clinical isolates and 4.07％of Gram-negative bacteria in CHINET program.Among the 40 437 strains of H.influenzae,66.89％were isolated from children and 33.11％were isolated from adults.More than 90％of the H.influenzae strains were isolated from respiratory tract specimens.The prevalence of β-lactamase was 53.79％in H.influenzae strains.The H.influenzae strains isolated from children showed higher resistance rate than the strains isolated from adults.Overall,779 strains of H.influenzae did not produce β-lactamase but were resistant to ampicillin(BLNAR).Beta-lactamase-producing strains showed significantly higher resistance rates to these antimicrobial agents than the β-lactamase-nonproducing strains.Of the 16 191 M.catarrhalis strains,80.06％were isolated from children and 19.94％isolated from adults.M.catarrhalis strains were mostly susceptible to both amoxicillin-clavulanic acid and cefuroxime,evidenced by resistance rate lower than 2.0％.Conclusions The emergence of antibiotic-resistant H.influenzae due to β-lactamase production poses a challenge for clinical anti-infective treatment.Therefore,it is very important to implement antibiotic resistance surveillance for H.influenzae and guide rational antibiotic use.All local clinical microbiology laboratories should actively improve antibiotic susceptibility testing and strengthen antibiotic resistance surveillance for H.influenzae.

Objective To investigate the distribution and antimicrobial resistance profiles of clinically isolated Haemophilus influenzae and Moraxella catarrhalis in hospitals across China from 2015 to 2021,and provide evidence for rational use of antimicrobial agents.Methods Data of H.influenzae and M.catarrhalis strains isolated from 2015 to 2021 in CHINET program were collected for analysis,and antimicrobial susceptibility testing was performed by disc diffusion method or automated systems according to the uniform protocol of CHINET.The results were interpreted according to the CLSI breakpoints in 2022.Beta-lactamases was detected by using nitrocefin disk.Results From 2015 to 2021,a total of 43 642 strains of Haemophilus species were isolated,accounting for 2.91％of the total clinical isolates and 4.07％of Gram-negative bacteria in CHINET program.Among the 40 437 strains of H.influenzae,66.89％were isolated from children and 33.11％were isolated from adults.More than 90％of the H.influenzae strains were isolated from respiratory tract specimens.The prevalence of β-lactamase was 53.79％in H.influenzae strains.The H.influenzae strains isolated from children showed higher resistance rate than the strains isolated from adults.Overall,779 strains of H.influenzae did not produce β-lactamase but were resistant to ampicillin(BLNAR).Beta-lactamase-producing strains showed significantly higher resistance rates to these antimicrobial agents than the β-lactamase-nonproducing strains.Of the 16 191 M.catarrhalis strains,80.06％were isolated from children and 19.94％isolated from adults.M.catarrhalis strains were mostly susceptible to both amoxicillin-clavulanic acid and cefuroxime,evidenced by resistance rate lower than 2.0％.Conclusions The emergence of antibiotic-resistant H.influenzae due to β-lactamase production poses a challenge for clinical anti-infective treatment.Therefore,it is very important to implement antibiotic resistance surveillance for H.influenzae and guide rational antibiotic use.All local clinical microbiology laboratories should actively improve antibiotic susceptibility testing and strengthen antibiotic resistance surveillance for H.influenzae.

Objective:To evaluate the efficacy and safety of remedial pedicled rectus abdominis myocutaneous flap transplantation for breast reconstruction.Methods:This retrospective study included 23 female breast cancer patients who underwent autologous breast reconstruction with a remedial pedicled rectus abdominis myocutaneous flap at Hunan Cancer Hospital from June 2022 to June 2023. The patients' ages ranged from 24 to 56 years, with a mean age of (43.2±1.3) years. All patients had previously undergone mastectomy and tumor resection before receiving the remedial pedicled rectus abdominis myocutaneous flap reconstruction. Follow-up was conducted through outpatient visits and telephone interviews, with a median follow-up period of 10 (6-15) months. Observation indicators included survival rate of flaps, patient satisfaction rate and aesthetic outcomes. Adverse reactions including but not limited to flap contracture and deformation, during and following the surgical procedure were also recorded.Results:The remedial pedicled rectus abdominis myocutaneous flap technique was utilized for autologous breast reconstruction in 23 patients. Specifically, this approach was chosen due to two primary reasons: the perforator distance from the vascular pedicle being excessively long in 18 cases, and suboptimal characteristics of the main pedicle in 5 cases. Two types of flap configurations were employed. For 15 patients, a unilateral remedial pedicled rectus abdominis myocutaneous flap was used, with an average flap size of (21.5±0.8) cm in length and (11.2±1.3) cm in width. All flaps demonstrated adequate blood supply without the need for additional vascular anastomosis. For the remaining 8 patients, a combination of a unilateral remedial pedicled rectus abdominis myocutaneous flap and a contralateral free lower abdominal flap was utilized, resulting in an average flap size of (29.3±0.7) cm in length and (12.0±1.8) cm in width. In these cases, the inferior epigastric vessels of the free lower abdominal flap were successfully anastomosed to the internal thoracic vessels of the recipient site, ensuring all flaps survived postoperatively. The survival rate of flaps in 23 patients was 100% (23/23), and the patient satisfaction rate was 100% (23/23). Postoperative evaluation revealed that the reconstructed breasts exhibited satisfactory shape and texture, with no instances of flap contracture or deformation. The reconstructed breasts were largely symmetrical with the contralateral side, leaving only linear scars at the donor and recipient sites. Importantly, there was no significant impact on abdominal wall function, and no adverse reactions such as flap contracture or impaired abdominal wall function were observed in any of the patients.Conclusion:When the characteristics of the perforating vessels in the lower abdomen are suboptimal, the use of a remedial pedicled rectus abdominis myocutaneous flap technique for breast reconstruction can ensure satisfactory outcomes and significantly enhance surgical safety.

ObjectiveThis study aims to investigate the mechanism in which Celastrus orbiculatus extract （COE） affects the proliferation and differentiation of gastric organoids and the expression of Lgr5 and thus reverses the precancerous lesions of gastric cancer （PLGC） by regulating the leucine-rich repeat containing G protein-coupled receptor 5 （Lgr5）/Wingless （Wnt）/β-catenin signaling pathway based on a gastric organoid injury model. MethodGastric organoids were established based on stem cells of the mouse gastric gland. Gastric organoid injury models were constructed by treating gastric organoids with N-methyl-N'-nitro-N-nitrosoguanidine （MNNG， 0.02 mg·L^-1）. Gastric organoid injury models were randomly divided into normal group， model group （0.02 mg·L^-1 MNNG）， low， medium， and high dose （5， 10， 20 mg·L^-1） groups of COE， and Wnt inhibitor Dickkopf-related protein 1 （DKK1） （0.5 mg·L^-1） group， and they were treated with respective agents for 24 h. The number and volume of gastric organoids under different drug concentrations were observed under a microscope. The viability of the gastric organoid injury models was detected by Methyl thiazolyl tetrazolium （MTT） assay. The morphology and pathology of gastric organoids were observed using Hematoxylin and Eosin （HE） staining. The expression levels of Lgr5， Mucin2 （MUC2）， Mucin5AC （MUC5AC）， Mucin6 （MUC6）， Wnt， and β-catenin in gastric organoids under different drug concentrations were detected by Western blot （WB）. ResultCompared with the normal group， the number， volume， and activity of gastric organoids in the model group were decreased （P<0.01）， while the expressions of Lgr5， MUC2， Wnt， and β-catenin were significantly increased （P<0.01）. The expressions of MUC5AC and MUC6 were significantly decreased （P<0.01）. Compared with the model group， the number and volume of gastric organoids in the low， medium， and high dose groups of COE were all improved （P<0.01）， and the vitality of gastric organoids was significantly enhanced （P<0.01）. The effect was the most significant at a COE concentration of 20 mg·L^-1 （P<0.01）. The expressions of Lgr5 and MUC2 in the medium and high dose groups of COE were significantly reduced （P<0.01）， while the expression of MUC5AC and MUC6 were significantly increased in the low， medium， and high dose groups of COE （P<0.05， P<0.01）. Compared with the model group， Wnt inhibitors could promote the expression of MUC5AC and MUC6 in gastric organoids （P<0.05， P<0.01） and reduce the expression of MUC2， Wnt， and β-catenin. In addition， the combined use of COE at high concentrations and Wnt inhibitors could further promote this trend （P<0.01）. ConclusionCOE inhibits the Wnt/β-catenin pathway by inhibiting the expression of Lgr5， MUC2， Wnt， and β-catenin and promoting the expression of MUC5AC and MUC6， thus promoting the proliferation and differentiation of gastric organoids and reversing the PLGC process.