ObjectiveThe exacerbating trend of global population aging poses profound socioeconomic and public health challenges, making the comprehensive elucidation of biological aging mechanisms and the discovery of effective anti-aging interventions an urgent priority in the life sciences. Based on our previous serum metabolomics findings that dimethylglycine, an intermediate metabolite of amino acid metabolism naturally present in the human body, was significantly enriched in the serum of longevity families, this study aimed to systematically investigate the anti-aging effects of dimethylglycine both in living organisms and in controlled laboratory environments, and to preliminarily elucidate its underlying molecular mechanisms. While existing literature indicates that dimethylglycine possesses antioxidant and immunomodulatory properties, its direct anti-aging efficacy and the specific molecular pathways through which it operates remain largely unexplored. MethodsTo comprehensively evaluate the anti-aging properties of dimethylglycine, we utilized replicative senescent human embryonic lung fibroblasts, specifically the WI-38 cell line, as an experimental model in a controlled laboratory environment. Cell viability and safety were thoroughly assessed using Cell Counting Kit-8 and lactate dehydrogenase release assays across various concentrations of dimethylglycine. The impact of dimethylglycine on cellular senescence phenotypes, oxidative stress, and proliferative capacity was evaluated via senescence-associated beta-galactosidase staining, reactive oxygen species fluorescence detection, and 5-ethynyl-2'-deoxyuridine incorporation assays. Furthermore, the molecular alterations of senescence-associated secretory phenotype factors and core senescence signaling pathways were quantified using quantitative reverse transcription polymerase chain reaction for the messenger RNA levels of interleukin-6, interleukin-8, p21, and matrix metalloproteinase-1, and enzyme-linked immunosorbent assay for the measurement of p16 and p21 protein expression levels. For the living organism model, the wild-type nematode Caenorhabditis elegans was used to evaluate systemic physiological effects. We conducted a comprehensive lifespan analysis at 20°C, heat stress resistance survival assays at 35℃, senescence-associated beta-galactosidase staining, lipofuscin accumulation tracking, intracellular reactive oxygen species measurement, and Oil Red O staining to ascertain systemic lipid accumulation. Additionally, network pharmacology bioinformatics tools, including PharmMapper and STRING databases, and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were utilized to predict target pathways, alongside highly detailed molecular docking simulations utilizing SwissDock and Protein-Ligand Interaction Profiler to examine interactions with the cytochrome P450 family 2 subfamily C member 9 protein. ResultsThe experimental outcomes robustly demonstrate the potent anti-aging capabilities of dimethylglycine. At the cellular level, toxicity analyses firmly confirmed that dimethylglycine is highly safe; continuous treatment with 50 mol/L and 70 mol/L of dimethylglycine for 5 d did not induce any cellular membrane damage or cytotoxicity, but rather actively promoted cellular proliferation. Utilizing the optimal standardized concentration of 50 mol/L, dimethylglycine treatment significantly ameliorated senescent phenotypic markers in human embryonic lung fibroblasts, which was evidenced by a drastic and highly significant reduction in the senescence-associated beta-galactosidase positive cell percentage (P<0.000 1) and intracellular reactive oxygen species levels (P<0.000 1), alongside a marked increase in the 5-ethynyl-2'-deoxyuridine-positive proliferation rate (P=0.003 5). On a molecular expression scale, dimethylglycine significantly downregulated the messenger RNA expression of multiple core senescence-associated secretory phenotype inflammatory factors, including interleukin-6, interleukin-8, p21, and matrix metalloproteinase-1. Concurrently, it effectively suppressed the protein expression of critical cell cycle arrest markers, diminishing p16 protein levels by 57.3% (P=0.000 4) and p21 protein levels by 27.2% (P=0.000 7). In the nematode Caenorhabditis elegans animal model, dimethylglycine significantly extended the mean lifespan from 20.402 d to an impressive 23.066 d (P<0.000 1) and notably enhanced overall survival rates under severe heat stress environmental conditions (P=0.017). Furthermore, systemic dimethylglycine intervention significantly mitigated age-related physiological decline by decreasing bodily lipofuscin accumulation (P<0.000 1), significantly reducing senescence-associated beta-galactosidase activity, lowering systemic reactive oxygen species fluorescence (P=0.008), and effectively alleviating overall fat accumulation (P<0.000 1). Mechanistically, extensive network pharmacology and Kyoto Encyclopedia of Genes and Genomes analyses strongly revealed that the potential targets of dimethylglycine are significantly enriched in fundamental drug metabolism and oxidative stress response pathways. Precision molecular docking simulations conclusively demonstrated that dimethylglycine forms highly stable structural interactions with the cytochrome P450 family 2 subfamily C member 9 protein, specifically highlighting the definitive formation of 5 stable hydrogen bonds involving serine 365, leucine 366, and serine 429 residues, as well as two critical salt bridge formations with arginine 97 and histidine 368 residues. It is additionally predicted to interact favorably with glutathione S-transferase family proteins. ConclusionDimethylglycine exhibits a profoundly significant and multifaceted anti-aging activity at both the cellular and entire living animal levels. By powerfully alleviating oxidative stress, heavily suppressing the core p16 and p21-dependent cellular senescence signaling pathways, and substantially mitigating the detrimental senescence-associated secretory phenotype, dimethylglycine effectively delays fundamental cellular senescence processes and drastically extends whole-organism lifespan. The biological mechanisms driving these robust protective effects are highly likely closely associated with its direct stable interactions with crucial metabolic and detoxifying enzyme systems, such as cytochrome P450 family 2 subfamily C member 9 and glutathione S-transferase family proteins, thereby systemically improving metabolic dysregulation and restoring critical redox homeostasis. This comprehensive study provides highly solid experimental evidence supporting dimethylglycine as a highly potent and safe potential anti-aging intervention agent, while simultaneously offering a clear molecular mechanistic explanation for the previously documented high abundance of dimethylglycine observed within exceptionally long-lived human populations.

ObjectiveTo investigate the mechanisms of Tongmai Yangxin pills （TMYX） and Tongxinluo capsules （TXL） in treating no-reflow （NR） after myocardial ischemia and reperfusion following the concept of treating the same disease with different methods， based on integrative pharmacology and experimental validation. MethodsEighty 8-week-old SPF-grade SD rats were randomly assigned into four groups （n=20）： sham operation， NR， TMYX （4 g·kg^-1）， and TXL （2 mg·kg^-1）. A rat model of myocardial ischemia-reperfusion no-reflow was established by in-situ ligation of the left anterior descending coronary artery. Gastric gavage was first performed 4 h after the operation， and samples were collected on day 7. Thioflavin S staining was used to observe the NR area in rat myocardium. Echocardiography was performed to examine the cardiac function. Hematoxylin-eosin （HE） staining was conducted to observe the pathological changes of the myocardial tissue. An automatic biochemical analyzer was adopted to measure myocardial enzyme activity. Then， integrated pharmacology was applied for Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis. Finally， Western blot was employed to quantify the protein levels of key targets in the myocardial tissue， including soluble guanylyl cyclase （sGC）， cyclic guanosine monophosphate （cGMP）/dependent protein kinase （PKG）， phosphorylated phosphatidylinositol 3-kinase （p-PI3K）， phosphorylated protein kinase B （p-Akt）， and hypoxia-inducible factor-1α （HIF-1α）. ResultsCompared with the sham operation group， the NR group showed increased NR area of myocardium， decreased left ventricular ejection fraction （EF）， left ventricular fractional shortening （FS）， left ventricular outflow tract peak velocity （LVOT Peak）， and left ventricular stroke volume （LVSV） （P<0.01）， fractured and disordered myocardial fibers as well as inflammatory cell infiltration in the myocardial tissue， enhanced activities of creatine kinase （CK）， creatine kinase isoenzyme （CK-MB）， and lactate dehydrogenase （LDH） in the myocardial tissue （P<0.01）， and downregulated protein levels of sGC， PKG， phosphorylated （p）-PI3K （Tyr458）， and p-Akt （Tyr315） in the myocardial tissue （P<0.05， P<0.01）. The expression level of HIF-1α protein showed a downward trend. Compared with the NR group， the TMYX group and TXL group exhibited a decreasing trend in myocardial NR area， EF， FS， and LVOT Peak significantly increased （P<0.05， P<0.01）， LVSV showed an upward trend， ameliorated myocardial pathological morphology and inflammatory infiltration， reductions in CK， CK-MB， and LDH activities （P<0.05， P<0.01）， and upregulated protein levels of sGC， PKG， p-PI3K （Tyr458） in myocardial tissue （P<0.05， P<0.01）， the protein expressions of p-Akt （Tyr315） and HIF-1α showed an upward trend. A comparison of the therapeutic effects between the two compound prescriptions showed that TMYX tended to exert a better effect in restoring cardiac function and protecting cardiac structure in NR rats， whereas TXL was more effective in reducing myocardial NR area and lowering myocardial enzyme activities in NR rats. Integrative pharmacology analysis combined with experimental verification demonstrated that both TMYX and TXL could alleviate NR through the following mechanisms： activating the cyclic cGMP/PKG signaling pathway to regulate vascular tone， activating the PI3K/Akt signaling pathway to dilate blood vessels， and activating the HIF-1 signaling pathway to inhibit oxidative stress. However， TMYX had an advantage in activating the cGMP/PKG pathway， while TXL was superior in activating the PI3K/Akt pathway. The two compound prescriptions exerted comparable effects on the HIF-1 signaling pathway， which might serve as their common therapeutic pathway. ConclusionBoth TMYX and TXL could alleviate NR damage. TMYX exerts its protective effect against NR mainly by activating the cGMP/PKG signaling pathway， while TXL exerts its effect mainly through the PI3K/Akt pathway. The HIF-1α signaling pathway may be a common pathway for the two compound prescriptions to exert their protective effects against NR. This study reveals the similarities and differences between TMYX and TXL in the treatment effect and mechanism for NR， providing an experimental basis and a theoretical basis for better clinical application of the two compound prescriptions.

Objective:To evaluate the influence of different positive end-expiratory pressure on the ventilation of laryngeal mask airway in pediatric patients undergoing laparoscopic surgery through ultrasound assessment.Methods:In this randomized controlled trial, 90 pediatric patients of both sexes, of American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ, aged 2-10 yr, with a body mass index of 12-22 kg/m 2, scheduled for elective laparoscopic surgery, were divided into 3 groups ( n=30 each) by a random number table method: group P0, group P3, and group P5. Each group adopted the pressure-controlled mode with an inspiration/expiration ratio of 1∶2, a flow rate of 2 L/min, and a respiratory rate of 16-26 breaths/min. The positive end-expiratory pressure was set at 0, 3 and 5 cmH 2O(1 cmH 2O=0.098 kPa) in P0, P3 and P5 groups respectively. The ultrasonic scores and misalignment rate of the laryngeal mask airway were recorded every 5 min following the pneumoperitoneum, and the occurrence of intraoperative hypoxemia (SpO 2 < 92%) and P ETCO 2 ≥ 45 mmHg(1 mmHg=0.133 kPa) was recorded. The development of hypoxemia, blood staining on the laryngeal mask airway and hoarseness after laryngeal mask airway removal were also recorded. Results:Compared with P0 group, the ultrasound scores were significantly increased at 15 min of pneumoperitoneum in P3 group ( P<0.05), and the ultrasound scores and the laryngeal mask airway misplacement rate were significantly increased, and no significant change was found in the other parameters in P5 group ( P<0.05). There were no statistically significant differences in each parameter between P5 group and P3 group ( P>0.05). There was no statistically significant difference in the incidence of postoperative complications among the three groups ( P>0.05). Conclusions:The laryngeal mask airway provides better ventilatory effect in pediatric patients undergoing laparoscopic surgery when the PEEP is set at 3 cmH 2O.

Objective:To investigate the clinical effect of reconstruction of the degloving injury of thumb and finger with free perforator flap of superficial peroneal artery.Methods:This is a retrospective study. From June 2020 to June 2023, 18 superficial peroneal artery perforator flaps from 14 calves were used to treat 15 degloved digital wounds of 14 patients in the Department of Hand Surgery, Suzhou Ruihua Orthopaedic Hospital. There were 13 single digital degloving wounds and 1 two-digital degloving wounds. Of which, 3 digits were reconstructed with 2 free perforator flaps of ipsilateral superficial peroneal artery and 12 with a single free perforator flap of superficial peroneal artery. The size of wounds was 4.0 cm×2.0 cm-10.0 cm×4.0 cm, and the flaps were 5.0 cm×3.0 cm-12.0 cm×4.0 cm in size. The donor sites in calves were sutured layer by layer with absorptive sutures. Postoperative follow-ups were conducted through regular outpatient visits, phone calls or WeChat. Survival of flaps, postoperative complications, therapeutic effect of the flaps and patient satisfaction were observed.Results:There was no vascular compromise or wound infection of the flaps. All patients were included in the 4 to 36 months of postoperative follow-up, with an average of 9.86 months. All flaps had good appearance, without obvious swelling. Colour and texture of the flaps was close to the surrounding skin. The flaps had no obvious pigmentation or ulcer and scar hyperplasia at recipient site. Sensation of the flaps recovered to S 2 to S 3. There was no obvious scar hyperplasia, pain or dysfunction at donor sites. According to the comprehensive evaluation scale of flap, the scores were found at 81 to 91, with an average score of 85 in the 14 patients, and of whom 2 were excellent and 12 were good. Patient satisfaction was evaluated according to the Michigan Hand Outcomes Questionnaire (MHQ) and 10 patients were very satisfied and 4 were satisfied. Conclusion:Free perforator flap of superficial peroneal artery is one of the ideal flaps in reconstruction of degloving injury of thumb and fingers. It features constant and multiple perforators, reliable blood supply, high survival rate, flexible in design, thin and a small damage to the donor site.

Laparoscopic subtotal cholecystectomy (LSC) has been a safe and viable alternative to conversion to laparotomy in cases of severe cholecystitis. The objective of this study is to determine the utility of intraoperative choledochoscopy in LSC for the exploration of the gallbladder, cyst duct, and subsequent stone clearance of the cystic duct in cases of severe cholecystitis. A total of 72 patients diagnosed with severe cholecystitis received choledochoscopy-assisted laparoscopic subtotal cholecystectomy (CALSC). A choledochoscopy was performed to explore the gallbladder cavity and/or cystic duct, and to extract stones using a range of techniques. The clinical records, including the operative records and outcomes, were subjected to analysis. No LSC was converted to open surgery, and no bile duct or vascular injuries were sustained. All stones within the cystic duct were removed by a combination of techniques, including high-frequency needle knife electrotomy, basket, and electrohydraulic lithotripsy. A follow-up examination revealed the absence of residual bile duct stones, with the exception of one common bile duct stone, which was extracted via endoscopic retrograde cholangiopancreatography. In certain special cases, CALSC may prove to be an efficacious treatment for the management of severe cholecystitis. This technique allows for optimal comprehension of the situation within the gallbladder cavity and cystic duct, facilitating the removal of stones from the cystic duct and reducing the residue of the non-functional gallbladder remnant.

Background: s/Aims: Non-invasive models stratifying clinically significant portal hypertension (CSPH) are limited. Herein, we developed a new non-invasive model for predicting CSPH in patients with compensated cirrhosis and investigated whether carvedilol can prevent hepatic decompensation in patients with high-risk CSPH stratified using the new model. Methods: Non-invasive risk factors of CSPH were identified via systematic review and meta-analysis of studies involving patients with hepatic venous pressure gradient (HVPG). A new non-invasive model was validated for various performance aspects in three cohorts, i.e., a multicenter HVPG cohort, a follow-up cohort, and a carvediloltreating cohort. Results: In the meta-analysis with six studies (n=819), liver stiffness measurement and platelet count were identified as independent risk factors for CSPH and were used to develop the new “CSPH risk” model. In the HVPG cohort (n=151), the new model accurately predicted CSPH with cutoff values of 0 and –0.68 for ruling in and out CSPH, respectively. In the follow-up cohort (n=1,102), the cumulative incidences of decompensation events significantly differed using the cutoff values of <–0.68 (low-risk), –0.68 to 0 (medium-risk), and >0 (high-risk). In the carvediloltreated cohort, patients with high-risk CSPH treated with carvedilol (n=81) had lower rates of decompensation events than non-selective beta-blockers untreated patients with high-risk CSPH (n=613 before propensity score matching [PSM], n=162 after PSM). Conclusions Treatment with carvedilol significantly reduces the risk of hepatic decompensation in patients with high-risk CSPH stratified by the new model.

Objective To optimize the processing technology of Curculigo orchioides Gaertn.Methods On the basis of single factor experiment,the ratio of material to liquid,frying temperature and frying time were taken as the influencing factors,and the comprehensive scores of appearance traits,curculigoside,orcinol glucoside and orcinol gentiobioside were taken as the evaluation indexes.The optimal processing technology was optimized by Box-Behnken response surface method combined with analytic hierarchy process,and the process verification was carried out.Results The optimal process was as follows:the raw C.orchioides pieces were mixed with the same amount of Evodia rutaecarpa juice,placed in a closed container,moistened until the juice was completely absorbed,stir-fried at 130 ℃ for 6 min,taken out and dried at 60 ℃.For each 100 kg of Curculigo,10 kg of Evodia rutaecarpa was used.Evodia rutaecarpa juice preparation method:10 kg of Evodia rutaecarpa was soaked in 12 times the amount of water for 60 min,decocted for 50 min,extracted for 3 times,combined with the filtrate,and concentrated to 100 kg to obtain Evodia rutaecarpa juice.Conclusion The optimal processing technology of Curculigo orchioides Gaertn.is stable and feasible.

Objective:To establish tumor specific protein (SP70) targeted tumor cell enrichment technology and to assess applicational value of next-generation sequencing (NGS) analysis for enriched circulating tumor cell (CTC) in precision medicines of non-small cell lung cancer (NSCLC).Methods:The monoclonal antibody NJ001 was covalently coupled to the surface of magnetic beads to build targeted magnetic bead enrichment technology based on SP70. The limit of detection, coincidence rate, interference experiment, recovery test and clinical performance were evaluated. From March 2016 to August 2017, NGS analysis with or without pre-treatment of targeted enrichment for serous fluids of 43 NSCLC in the First Affiliated Hospital with Nanjing Medical University were compared (Kappa or Fisher exact test).Results:The CTC enrichment technology based on SP70 targeted immunomagnetic beads can specifically enrich tumor cells. The limit of detection was 10 4 SPC-A1 cells/L, and the coincidence rate, sensitivity and specificity were 100% (3/3). The endogenous interfering substances such as red blood cells, hemoglobin, white blood cells, epithelial cells and triglycerides had no interfering effects, as well as the exogenous interfering substances such as EDTA-K2, cefoxitin, carboplatin and paclitaxel. The recovery rate was 56.0% (56 000/100 000). A total of 30 gene mutations including 65 loci were found in 43 NSCLC under SP70 targeted enrichment, with a higher detection rate compared with unenrichment method [95.0% (19/20) vs 65.0% (13/20), χ 2=5.625, P=0.044]. Conclusion:In this study, SP70-targeted enriched CTC liquid biopsy method was established, with higher sensitivity and specificity of NGS detection than unenrichment method.

Biofilm formation is one of the key reasons that bacterial infections are difficult to treat.So it is of great significance to develop effective strategies to resist bacterial biofilms.Although antibiotics are important clinical tools for the treatment of bacterial infections,their therapeutic efficacy is often unsatisfactory when targeting bacterial biofilm-associated diseases.In this study,exopolysaccharides(EPS)from Escherichia coli(E.coli)were extracted and purified.It was demonstrated that the obtained E.coli EPS had the ability to inhibit methicillin-resistant Staphylococcus aureus(MRSA)biofilm formation and disperse a mature biofilm.To improve the anti-biofilm effect of vancomycin(VAN),E.coli EPS was used in combination with VAN.The combination increased the inhibition rate of MRSA biofilm and increased the dispersion rate of mature biofilm from 10%to 80%.When combined with E.coli EPS,the number of bacterial colonies within the MRSA biofilm remarkably decreased by 88%,resulting in a significant improvement over the use of VAN alone at an equivalent concentration.Meanwhile,E.coli EPS could down-regulate the expression of MRSA biofilm-related genes.E.coli EPS showed good anti-biofilm effect,and E.coli EPS/VAN combination could provided a potential strategy for treatment of MRSA biofilm infections.