Objective To explore the relationship between the inhibitory effect of quercetin on AC16 cardiomyocyte proliferation and the Wnt/β-catenin signaling pathway,the relationship between quercetin(quercetin,QCT)and the proliferation of AC16 cardiomyocytes through in vitro was investigated.Methods AC16 cells were stimulated with different concentrations of QCT.The effects of QCT on AC16 cell proliferation were detected by inverted microscope photography,trypan blue counting,and CCK-8 assay.Western blot was used to detect the effects of QCT on the expression of phosphorylated β-catenin(p-β-catenin),c-Myc,β-catenin,low density lipoprotein receptor-related protein 5(LRP5),and LRP6.The effect of quercetin on cell proliferation was detected after overexpressing β-catenin ΔN,LRP5,and LRP6 genes,and after silencing LRP5 and LRP6 genes by trypan blue counting.Results Compared with the control group,QCT could decrease the number of AC16 cells and inhibit the proliferation rate,which was concentration-dependent.At the protein expression level,10 and 20 μmol/L QCT led to an significant upregulated modification of p-β-catenin protein(P<0.05)and significant downregulation of c-Myc,β-catenin,LRP5,and LRP6 protein expression(P<0.05)in AC16 cells.Therefore,10 μmol/L QCT was chosen as the intervention concentration.Overexpression of β-catenin ΔN,LRP5,and LRP6 genes in AC16 cells significantly rescued the cell proliferation inhibition caused by 10 μmol/L QCT compared to the drug-only group(P<0.05).Conversely,silencing LRP5 and LRP6 genes led to inhibition of AC16 cell proliferation,and the combination with 10 μmol/L QCT did not exacerbate the inhibition(P>0.05).Conclusion The quercetin could inhibit the Wnt/β-catenin signaling pathway via significant downregulation of LRP5/6,thereby attenuate cell proliferation of AC16 cardiomyocytes.

In recent years,organs-on-chips(OOCs)technology has gradually matured and been recognized by regulatory agencies,playing a key role in the construction of pathophysiological models,pharmacodynamic screening,toxicological evaluation and other fields,providing guidance for the research and development of innovative drugs of Chinese materia medica and the clinical application of classic prescriptions.This article reviewed the application progress of OOCs technology in the research of Chinese materia medica from four dimensions:pharmacodynamic and toxicological evaluation,quality control,active component analysis and innovative drug research and development,in order to provide a reference for the precise application of this technology in the field of Chinese materia medica.

Objective:To explore the application value of transpubic access on female urogenital tract injury.Methods:A retrospective analysis was conducted on 15 female patients with urogenital tract injury caused by trauma admitted to our department from May 2020 to October 2024，all of whom were complicated urethral stricture or atresia，accompanied by urethrovaginal fistula or vaginorectal fistula. All patients underwent suprapubic vesicostomy before surgery，and 1 case underwent sigmoidostomy simultaneously. The mean age of the patients was（29.6 ± 3.2）years old，and the course of disease was 6-24 months. Preoperative urethrography and urethroscope showed the location of urethral stenosis，with proximal urethra stricture in 7 cases and distal urethra stricture in 8 cases. The average length of strictures was（2.8±0.2）cm. The urethral ultrasonography，magnetic resonance and CTU examination showed 8 patients were complicated with urethrovaginal fistula，and 1 patient was complicated with vaginorectal fistula. All patients underwent transpubic access and resection of symphysis pubis. According to the specific conditions of urethral stricture，7 of them underwent end-to-end urethral anastomosis，5 cases underwent bladder wall flap urethroplasty，3 cases underwent vulva flap urethroplasty，8 cases underwent urethral vaginal fistula repair，1 case underwent vagino-rectal fistula repair，and 7 cases underwent vaginoplasty during the operation.Results:All the 15 patients underwent successful operation without complication. After the catheter being removed 4 weeks after surgery，2 patients had urgent urinary incontinence and 3 patients had stress urinary incontinence. The bladder neck was reconstructed 3 months after surgery，the symptoms of urinary incontinence improved in 1 case，urinary incontinence remained in 2 cases，and pharmaceutical or physical therapy were continued. Two patients could not urinate normally after the catheter was removed and still carried the vesicostomy tube，waiting for further treatment. The other 8 patients had unobtrusive voiding after extubating，and were followed up for an average of（22.5±3.2）months. There was no recurrence of urinary fistula，and the average maximum urinary flow rate was（22.8±3.2）ml/s.Conclusions:The transpubic approach is a safe and effective way to treat female genital tract injury by different surgical methods according to specific conditions，especially for patients with severe trauma，poor local tissue conditions，complicated urethrovaginal fistula or vagino-rectal fistula.

BACKGROUND:The technique of ultrasound processing is widely used for molecular biological analysis.It is of great significance to study the DNA quality of tissue with different storage years under new ultrasonic treatment for further specimen quality control of molecular detection.OBJECTIVE:To explore the effects of different storage durations on DNA quality in specimens with ultrasound processing to investigate the optimal storage time for molecular tests.METHODS:Forty specimens of breast biopsy were collected and paraffin specimens were prepared by ultrasonography.These specimens were divided into four groups based on their storage periods:<1 year,1-3 years,>3-5 years,and>5 years,which contained 10 cases in each group.Paraffin specimens were sliced;each slice was 3 μm thick;10-15 slices were taken,and DNA was extracted.The mass concentration of DNA was examined by Nanophotometer N60 ultra-micro spectrophotometer and Qubit 4.0 fluorometer.The purity of the DNA was analyzed by the ratio of A260/A280.DNA fragment integrity was measured by capillary electrophoresis (Qsep 100) to evaluate the quality of the DNA fragments.RESULTS AND CONCLUSION:The mean values of A260/A280 in the four groups were between 1.8 and 2.0,meeting the requirements of tests,without significant differences.The mean values of DNA mass concentration (Qubit concentration) were 30.39,14.33,2.52,and 1.95 ng/μL,respectively.The mean values of the N/Q were 6.48,14.18,24.56,and 29.86.The mean values of DNA were:5.64,1.76,1.24,and 0.80.The percentage of large DNA fragments averaged 56.08％,17.72％,12.68％,and 7.90％.Moreover,the Ct values of the internal control detected by PCR were 15.32,17.09,18.39,and 21.24.The three other groups exhibited significantly lower DNA concentration,higher N/Q ratios,decreased DNA quality and percentage of large fragments,and increased values of Ct,compared with the group of within 1 year of storage (P<0.05).The experimental results suggested that for novel ultrasound processed biopsy specimens,we should prioritize samples stored within 1 year for molecular testing.Samples stored within 3 years can also meet the requirements of second-generation sequencing and other tests.Samples stored within 5 years can only be attempted to carry out PCR.Samples stored for more than 5 years were not recommended to carry out molecular tests.

Objective:To explore the application value of transpubic access on female urogenital tract injury.Methods:A retrospective analysis was conducted on 15 female patients with urogenital tract injury caused by trauma admitted to our department from May 2020 to October 2024，all of whom were complicated urethral stricture or atresia，accompanied by urethrovaginal fistula or vaginorectal fistula. All patients underwent suprapubic vesicostomy before surgery，and 1 case underwent sigmoidostomy simultaneously. The mean age of the patients was（29.6 ± 3.2）years old，and the course of disease was 6-24 months. Preoperative urethrography and urethroscope showed the location of urethral stenosis，with proximal urethra stricture in 7 cases and distal urethra stricture in 8 cases. The average length of strictures was（2.8±0.2）cm. The urethral ultrasonography，magnetic resonance and CTU examination showed 8 patients were complicated with urethrovaginal fistula，and 1 patient was complicated with vaginorectal fistula. All patients underwent transpubic access and resection of symphysis pubis. According to the specific conditions of urethral stricture，7 of them underwent end-to-end urethral anastomosis，5 cases underwent bladder wall flap urethroplasty，3 cases underwent vulva flap urethroplasty，8 cases underwent urethral vaginal fistula repair，1 case underwent vagino-rectal fistula repair，and 7 cases underwent vaginoplasty during the operation.Results:All the 15 patients underwent successful operation without complication. After the catheter being removed 4 weeks after surgery，2 patients had urgent urinary incontinence and 3 patients had stress urinary incontinence. The bladder neck was reconstructed 3 months after surgery，the symptoms of urinary incontinence improved in 1 case，urinary incontinence remained in 2 cases，and pharmaceutical or physical therapy were continued. Two patients could not urinate normally after the catheter was removed and still carried the vesicostomy tube，waiting for further treatment. The other 8 patients had unobtrusive voiding after extubating，and were followed up for an average of（22.5±3.2）months. There was no recurrence of urinary fistula，and the average maximum urinary flow rate was（22.8±3.2）ml/s.Conclusions:The transpubic approach is a safe and effective way to treat female genital tract injury by different surgical methods according to specific conditions，especially for patients with severe trauma，poor local tissue conditions，complicated urethrovaginal fistula or vagino-rectal fistula.

Liu Baoyi was a famous febrile disease expert in the late Qing Dynasty,who innovated the theory of latent febrile disease.Liu proposed that the deficiency of kidney essence and the deficiency of healthy qi were the prerequisites for the latent cold e-vil,and the exogenous cold evil was the inducement of the disease.He advocated the treatment of latent febrile disease based on the six meridians,with the two ideas of eliminating evil qi and strengthening healthy qi as the outline,and the general principle of treatment was to relieve heat,nourish yin and expel evil qi.He attached importance to the coordination of medicine and food,made good use of fresh medicine,attached importance to drug processing,emphasized the use of herb processed by herb,and adapted the medicine ac-cording to the syndromes.He preferred to use co-processed products for clinical use.The study of Liu's academic thoughts and medi-cation characteristics in treating latent febrile diseases is of great value in guiding the treatment and medication of various infectious and epidemic diseases with fever characteristics nowadays.

Objective:To evaluate the feasibility and preventive efficacy of a fascia- locking circular continuous suture ostomy technique in reducing parastomal hernia incidence.Methods:This technique was applied to patients undergoing permanent colostomy following radical rectal cancer resection. Surgical steps included: (1) A circular incision was made 1-2 cm medial to the intersection of the lateral margin of the rectus abdominis muscle and the line connecting the umbilicus to the left anterior superior iliac spine. Subcutaneous tissues were dissected vertically to expose the anterior rectus sheath, followed by blunt separation of the rectus abdominis after longitudinal incision of the sheath. The posterior rectus sheath and peritoneum were similarly incised. (2) Eight equidistant interrupted sutures (anchoring knots) were placed through the anterior rectus sheath, partial rectus abdominis, posterior rectus sheath, and peritoneum. (3) The terminal colon was exteriorized, and continuous sutures were applied to secure the anchoring knots and seromuscular layers of the bowel between knots, forming a circular locking mechanism by tying the terminal suture to the initial knot's tail. (3) The skin and seromuscular layers of the bowel margin were intermittently sutured (8-12 stitches) to achieve mucosal eversion.Results:From February to October 2023, 13 patients (11 males, 2 females; age: 67 ± 10 years; BMI: 23.8 ± 4.0 kg/m2) underwent this technique at the Second Affiliated Hospital of Army Medical University. Mean stoma creation time was 15.7 ± 3.0 minutes. During a follow-up of 14.6 ± 3.1 months, physical examinations and abdominal CT scans identified parastomal hernias in 2 male patients at 10 and 7 months postoperatively. Only one patient experienced a Clavien-Dindo grade ≥Ⅲ complication, which resolved with treatment. No stoma-related complications (e.g., infection, stenosis, or prolapse) occurred in any patient.Conclusion:The fascia-locking circular continuous suture ostomy technique is safe and feasible, demonstrating potential efficacy in preventing parastomal hernia following colostomy.

Background and Aims:N6-methyladenosine(m6A)epigenetic modification plays a crucial role in post-transcriptional gene expression regulation and various physiological and pathological processes,including tumorigenesis.The m6A reader IGF2BP2 significantly enhances mRNA stability and translation efficiency and is abnormally expressed in multiple cancers.However,the specific biological function of IGF2BP2 in pancreatic cancer remains unclear.Therefore,this study investigated the expression of the m6A reader IGF2BP2 in pancreatic cancer and its effects on pancreatic cancer cell functions.Methods:The expression levels of m6A-related writers,erasers,and readers were analyzed using The Cancer Genome Atlas(TCGA),the Genotype-Tissue Expression(GTEX)database,and the Gene Expression Omnibus(GEO).Kaplan-Meier survival analysis was conducted to assess the relationship between IGF2BP2 expression and the prognosis of pancreatic cancer patients.Immunohistochemistry was used to validate IGF2BP2 expression in clinical specimens of pancreatic cancer tissues and adjacent normal tissues.Functional experiments,including CCK-8 assay,flow cytometry for cell cycle analysis,colony formation assay,and Transwell migration assay,were performed to evaluate changes in cell proliferation,cell cycle distribution,colony formation ability,and migration capacity after IGF2BP2 knockdown in pancreatic cancer cells.Results:TCGA-GTEX and GEO database analyses showed that IGF2BP2 was highly expressed in pancreatic cancer tissues(both P<0.05)and that its high expression was associated with poor overall survival(both P<0.05).Immunohistochemical staining of clinical specimens confirmed that IGF2BP2 protein expression was higher in pancreatic cancer than in adjacent normal tissue.Functional experiments demonstrated that IGF2BP2 knockdown significantly reduced the proliferation ability of pancreatic cancer cells,arrested more cells in the G0-G1 phase,decreased colony formation,and impaired cell migration(all P<0.05).Conclusion:The m6A reader IGF2BP2 is highly expressed in pancreatic cancer tissues and is closely associated with poor prognosis in patients with this disease.Its mechanism of action may be related to the promotion of cancer cell growth and migration.

Objective To explore the relationship between the inhibitory effect of quercetin on AC16 cardiomyocyte proliferation and the Wnt/β-catenin signaling pathway,the relationship between quercetin(quercetin,QCT)and the proliferation of AC16 cardiomyocytes through in vitro was investigated.Methods AC16 cells were stimulated with different concentrations of QCT.The effects of QCT on AC16 cell proliferation were detected by inverted microscope photography,trypan blue counting,and CCK-8 assay.Western blot was used to detect the effects of QCT on the expression of phosphorylated β-catenin(p-β-catenin),c-Myc,β-catenin,low density lipoprotein receptor-related protein 5(LRP5),and LRP6.The effect of quercetin on cell proliferation was detected after overexpressing β-catenin ΔN,LRP5,and LRP6 genes,and after silencing LRP5 and LRP6 genes by trypan blue counting.Results Compared with the control group,QCT could decrease the number of AC16 cells and inhibit the proliferation rate,which was concentration-dependent.At the protein expression level,10 and 20 μmol/L QCT led to an significant upregulated modification of p-β-catenin protein(P<0.05)and significant downregulation of c-Myc,β-catenin,LRP5,and LRP6 protein expression(P<0.05)in AC16 cells.Therefore,10 μmol/L QCT was chosen as the intervention concentration.Overexpression of β-catenin ΔN,LRP5,and LRP6 genes in AC16 cells significantly rescued the cell proliferation inhibition caused by 10 μmol/L QCT compared to the drug-only group(P<0.05).Conversely,silencing LRP5 and LRP6 genes led to inhibition of AC16 cell proliferation,and the combination with 10 μmol/L QCT did not exacerbate the inhibition(P>0.05).Conclusion The quercetin could inhibit the Wnt/β-catenin signaling pathway via significant downregulation of LRP5/6,thereby attenuate cell proliferation of AC16 cardiomyocytes.

Objective To investigate the predictive value of the combined tumor burden score (TBS) and platelet-albumin-bilirubin (PALBI) score model for postoperative tumor recurrence in liver transplant recipients with hepatocellular carcinoma (HCC). Methods The general information of 158 recipients diagnosed with HCC and underwent liver transplantation at the 900th Hospital of the Joint Logistics Support Force of the Chinese People's Liberation Army from 2008 to 2021 was collected. Lasso regression analysis combined with multivariate Cox regression analysis were used to identify independent risk factors for postoperative tumor recurrence after liver transplantation with HCC. A nomogram prediction model was constructed based on variables selected by Lasso regression analysis, and the predictive performance of the model was verified by calibration curve and clinical decision curve. The optimal cut-off values for postoperative tumor recurrence in liver transplant recipients with HCC were determined by receiver operating characteristic (ROC) curve, and Kaplan-Meier analysis was used to compare survival differences among different groups. Results Among the 158 liver transplant recipients with HCC, 82 experienced tumor recurrence, with a recurrence rate of 51.9% and a median tumor-free survival time of 10 (4, 25) months. Results of Lasso regression analysis and multivariate Cox regression analysis showed that alpha-fetoprotein (AFP) ≥400 ng/mL, TBS and PALBI score were all independent risk factors for postoperative tumor recurrence in liver transplant recipients with HCC (all P<0.05). The combined high TBS-high PALBI score showed the highest predictive value (hazard ratio 6.909, 95% confidence interval 3.067-15.563, P<0.001). A nomogram prediction model was constructed based on six variables selected by Lasso regression analysis. Calibration curve showed good consistency between the model's predicted results and the ideal curve. Decision curve analysis indicated that the nomogram prediction model provided the highest clinical benefit for predicting 1-year tumor-free survival after liver transplantation with HCC. Time-dependent ROC curves at 1, 3 and 5 years after surgery showed that TBS-PALBI model had good predictive performance, with no significant difference in area under the curve (AUC) compared with TBS-PALBI-AFP model. The optimal cut-off values for predicting postoperative tumor recurrence were determined by ROC curve, with a PALBI score cut-off of −2.334 and a TBS cut-off of 5.305. Recipients were divided into a low TBS-low PALBI score group (n=47) and a low/high TBS-low/high PALBI score group (at least one score was high) (n=111). Kaplan-Meier survival analysis showed that the low TBS-low PALBI score group had a higher tumor-free survival rate than the low/high TBS-low/high PALBI score group, with a significant difference (P<0.05). Conclusions TBS-PALBI model provides a novel, simple and effective tool for assessing the prognosis of liver transplant recipients with HCC. The nomogram model constructed based on this has significant advantages in predictive performance and may serve as a reference for guiding individualized treatment plans and improving clinical outcomes.