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.

Puerariae Lobatae Radix and Puerariae Thomsonii Radix are the dried roots of Pueraria lobata and Pueraria thomsonii of the Leguminosae family， respectively. Since their separate inclusion in the 2005 edition of Pharmacopoeia of the People's Republic of China， the two herbs have been individually recorded. Although they share the same functions and indications， the required content of puerarin differs by a factor of up to eightfold between them. The current quality standard relies solely on a single indicator， puerarin， which fails to reflect the correlation between chemical constituents and core therapeutic effects. This has resulted in prominent issues of mixed use in clinical practice and production， thereby constraining the high-quality development of the industry. Modern research has demonstrated that Puerariae Lobatae Radix contains a comprehensive range and higher content of isoflavonoid constituents， while Puerariae Thomsonii Radix is rich in starch and polysaccharides. Puerariae Lobatae Radix exhibits stronger effects in antipyresis， cardiovascular and cerebrovascular protection， and central analgesia， while Puerariae Thomsonii Radix demonstrates greater advantages in anti-inflammation， hypoglycemia， and peripheral analgesia. Based on this， the chemical material bases of Puerariae Lobatae Radix and Puerariae Thomsonii Radix at both the macromolecule and micromolecule levels were systematically reviewed and compared. The pharmacodynamic material bases and mechanisms of action underlying their three core therapeutic effects， namely "relieving exterior syndrome and clearing heat"， "promoting fluid production to quench thirst"， and "dredging channels and activating collaterals"， were summarized. The most highly correlated common metabolic pathway shared by both herbs for these three therapeutic effects is the arachidonic acid metabolic pathway， and their common core target is prostaglandin-endoperoxide synthase 2 （PTGS2）. This review provided clinical application recommendations based on the therapeutic differences between the two herbs， elucidated the scientific connotation of their "same origin but different effects"， and， grounded in the quality marker （Q-Marker）， proposed the concept of efficacy-related Q-Marker based on "substance-efficacy-syndrome" association， as well as a novel strategy for efficacy-oriented quality evaluation. Preliminary screening of quality markers associated with different therapeutic effects was also conducted. This study aims to provide critical scientific evidence for the precise clinical application of Puerariae Lobatae Radix and Puerariae Thomsonii Radix， the improvement of quality standards， and the high-quality development of the industry.

ObjectiveTo investigate the impact of a cold environment on the pathophysiological changes in dogs after abdominal gastrointestinal blast injury. MethodsEight healthy common-grade adult male Beagles were selected and randomly divided into a cold group and a normal temperature group, with 4 dogs in each group. Prior to surgery, Beagles dogs were fasted and deprived of water for 12 hours; after establishing a vein at the edge of the ear, propofol （2 mg·kg^-1·min^-1） was continuously infused using a microinfusion pump to maintain anesthesia. Tracheal intubation was performed while preserving spontaneous breathing, and the electroencephalogram bifrequency index was maintained at 40-60 to ensure stable anesthesia. A model of abdominal gastrointestinal blast injury in Beagle dogs was established using a multifunctional combat trauma modeling platform. After blast injury, the cold group was exposed to a cold chamber at -20 ℃ combined with 10% body surface area ice water immersion for 40 min, while the normal temperature group was kept in an environment at (25±2) ℃ for 40 min after blast injury. Vital signs such as heart rate, blood pressure, and blood oxygen saturation were monitored in both groups before blast injury, immediately after blast injury, and 40 min after blast injury. And arterial blood gas parameters, including pH, partial pressure of oxygen in arterial blood (PaO₂), partial pressure of carbon dioxide in arterial blood (PaCO₂), actual bicarbonate radical (HCO₃^-), lactic acid (Lac), and blood base excess (BEb), were measured. After the experiment, gastrointestinal tissues (gastric antrum and upper jejunum 5 cm distal to the suspensory ligament of the duodenum) were harvested for pathological analysis via hematoxylin-eosin (HE) staining combined with semi-quantitative scoring. ResultsPhysiological indicators showed that the body temperature of the cold group was significantly lower than that of the normal temperature group (P<0.01), and the heart rate and systolic blood pressure of the cold group were significantly higher than those of the normal temperature group (P<0.000 1). PaCO₂ was markedly elevated and BEb was markedly decreased in the cold group relative to the normal temperature group (P<0.000 1), suggesting aggravated respiratory acidosis and metabolic acidosis. Pathological analysis showed that gastro-intestinal tissue damage was more severe in the cold group: the gastric mucosal epithelial shedding rate was > 60% (vs. <25% in the normal temperature group), the necrosis rate of the epithelial cells at the tips of intestinal villi reached more than 70% (vs. < 30% in the normal temperature group), and inflammatory cell infiltration involved the submucosa (vs. limited to the superficial mucosa in the normal temperature group). The semi-quantitative scoring results were consistent with the pathological analysis results, further confirming that the cold environment exacerbated gastrointestinal tissue damage caused by abdominal gastrointestinal blast injury. ConclusionA cold environment can exacerbate circulatory dysfunction and gastrointestinal tissue damage in Beagle dogs after blast injury, providing experimental evidence for early rewarming intervention for combat trauma in cold regions.

ObjectiveTo develop an early - warning approach for influenza in Anhui province using the moving epidemic method （MEM）， offering a scientific basis for influenza prevention and control in the province. MethodsBased on the surveillance data reported weekly by influenza sentinel hospitals and network laboratories in the “China Influenza Surveillance Information System”， the weekly influenza virus detection positive rate （positive rate， PR） across the province was analyzed. The MEM was used to estimate the epidemic threshold and classification intensity threshold of influenza in Anhui province before and after the COVID-19 pandemic， respectively. The early warning effect of the MEM was evaluated using cross-validation. ResultsEpidemic and graded intensity thresholds of influenza were derived using PR and MEM. Prior to the COVID-19 pandemic， the starting， medium-intensity， high-intensity， extremely-high-intensity， and ending thresholds of influenza epidemics were 17.08%， 38.47%， 53.20%， 59.71%， and 12.34% respectively. At this point， the model exhibited sensitivity of 0.98， specificity of 0.86， Matthews correlation coefficient of 0.84， and Youden index of 0.84. After the COVID-19 pandemic， these thresholds were 25.61%， 43.81%， 63.29%， 74.46%， and 17.82% respectively. At this point， the model demonstrates sensitivity of 0.86， specificity of 0.92， Matthews correlation coefficient of 0.78， and Youden index of 0.77. ConclusionMEM shows good applicability in estimating the epidemic and graded intensity thresholds of influenza during the winter and spring in Anhui province. Both the influenza epidemic threshold and graded intensity thresholds after the COVID-19 pandemic are higher than those before the pandemic.

Background: Maturity-onset diabetes of the young (MODY) due to variants of hepatocyte nuclear factor 1-beta (HNF1β) (MODY5) has not been well studied in the Chinese population. This study aimed to estimate its prevalence and evaluate the application of a clinical screening method (Faguer score) in Chinese early-onset diabetes (EOD) patients. Methods: Among 679 EOD patients clinically diagnosed with type 2 diabetes mellitus (age at diagnosis ≤40 years), the exons of HNF1β were sequenced. Functional impact of rare variants was evaluated using a dual-luciferase reporter system. Faguer scores ≥8 prompted multiplex ligation-dependent probe amplification (MLPA) for large deletions. Pathogenicity of HNF1β variants was assessed following the American College of Medical Genetics and Genomics (ACMG) guidelines. Results: Two rare HNF1β missense mutations (E105K and G454R) were identified by sequencing in five patients, showing functional impact in vitro. Another patient was found to have a whole-gene deletion by MLPA in 22 patients with the Faguer score above 8. Following ACMG guidelines, six patients carrying pathogenic or likely pathogenic variant were diagnosed with MODY5. The estimated prevalence of MODY5 in Chinese EOD patients was approximately 0.9% or higher. Conclusion MODY5 is not uncommon in China. The Faguer score is helpful in deciding whether to perform MLPA analysis on patients with negative sequencing results.

Nitrogen dioxide(NO 2)is a prevalent air pollutant that poses significant threats to the environment and human health,emphasizing the urgent need for high-performance NO 2 sensors for effective environmental monitoring at room temperature.Sensors based on metal halide perovskites have emerged as promising candidates for gas detection at room temperature,but their long-term stability remains a major challenge.In this study,a methylamine thiocyanate(MT)-doped FA 0.8Cs 0.2PbI 2Br(PVK)gas sensor(MT-PVK)was prepared using a simple one-step spin-coating method and ethyl acetate(EA)anti-solvent extraction technique.The crystalline structure,chemical composition,and particle morphology of the MT-PVK thin film were characterized.The MT-PVK thin film was then utilized as a gas sensor for NO 2 detection at room temperature.The results demonstrated that the sensor exhibited outstanding selectivity and reversibility at low concentrations of NO 2 gas,with a detection limit as low as 33 ppb(10-9,nL/L).For 10 ppm(10-6,μL/L)NO 2,the sensor showed rapid response and recovery time of only 1.6 s and 27 s,outperforming most traditional metal oxide NO 2 sensors.Furthermore,compared to the original PVK,the MT molecules significantly enhanced the structural and sensing stability of the MT-PVK sensor under high humidity conditions(55%±5%).These findings suggested that MT-doped FA 0.8Cs 0.2PbI 2Br perovskite gas sensors offered a promising pathway for the development of rapid-response gas sensing technologies suitable for room temperature operation.

Objective To investigate the expression of microRNA-873-5p(miR-873-5p)and C-X-C motif chemokine ligand 16(CXCL16)in thyroid cancer tissue and their relationship with pathological parameters and prognosis.Methods A total of 125 patients with thyroid cancer who underwent surgery at Nanjing Sec-ond Hospital from January 2018 to June 2019 were selected as the research subjects.Some cancer tissues and corresponding adjacent tissues of the patients were collected,and the expressions of miR-873-5p and CXCL16 mRNA were detected by real-time fluorescence quantitative polymerase chain reaction.The binding sites of miR-873-5p and CXCL16 were predicted through the online database.Pearson correlation was used to analyze the correlation between miR-873-5p and CXCL16 mRNA expression,and the correlation between miR-873-5p,CXCL16 mRNA expression and pathological parameters.According to the median expression of miR-873-5p and CXCL16 mRNA in thyroid cancer tissues,they were classified as high expression and low expression.The survival curves of patients with high and low expression of miR-873-5p and CXCL16 mRNA were plotted by the Kaplan-Meier method.Taking the survival status of patients with thyroid cancer as the dependent varia-ble,Cox regression was used to determine the relationship between the expressions of miR-873-5p and CX-CL16 mRNA and the death of patients with thyroid cancer.Results The expressions of miR-873-5p and CX-CL16 mRNA in thyroid cancer tissues were 0.83±0.12 and 1.54±0.25,respectively,and those in adjacent tissues were 1.13±0.15 and 0.98±0.13,respectively,the differences were statistically significant(t=-18.160,21.089).P＜0.001).Pearson correlation analysis showed that the expression of miR-873-5p and CXCL16 mRNA in thyroid cancer tissues was negatively correlated(r=-0.722,P＜0.001).The expression of miR-873-5p in thyroid cancer tissues was negatively correlated with pathological type,TNM stage and lymph node metastasis(r=-0.510,—0.262,-0.315,P＜0.05).The expression of CXCL16 mRNA was positively correlated with pathological type,TNM stage and lymph node metastasis(r=0.593,0.275,0.314,P＜0.05).The Kaplan-Meier survival curve showed that the 5-year overall survival rate of patients with high expression of miR-873-5p was higher than that of patients with low expression of miR-873-5p.The 5-year o-verall survival rate of patients with high expression of CXCL16 mRNA was lower than that of patients with low expression of CXCL16 mRNA,and the difference was statistically significant(x2=11.328,10.514,all P=0.001).miR-873-5p≥0.84 was an independent protective factor for death in patients with thyroid cancer,and CXCL16 mRNA≥1.55 was an independent risk factor for death in patients with thyroid cancer(P＜0.05).Conclusion The expressions of miR-873-5p and CXCL16 mRNA in thyroid cancer tissues are related to patho-logical parameters and prognosis,and may become prognostic markers for patients with thyroid cancer.

Objective To explore the relationship between systemic immune-inflammation index(SII),red blood cell distribution width(RDW),25-hydroxy-vitamin-D[25(OH)D]levels and frailty index in elderly pa-tients with type 2 diabetes mellitus(T2DM).Methods A total of 197 elderly patients with T2DM admitted to the hospital from March 2023 to March 2024 were collected as the research subjects.The patients were divided into the frailty group(106 cases)and the non-frailty group(91 cases)according to the scores of the clinical frailty scale.The clinical data and the levels of SII,RDW and 25(OH)D of the two groups were compared.Pearson correlation analysis was used to analyze the correlations between the levels of SII,RDW and 25(OH)D and the frailty index of elderly patients with T2DM.Logistic regression was used to analyze the influencing factors of frailty in elderly patients with T2DM.Results Compared with the non-frailty group,the proportion of women,the history of falls within 1 year,and the age of the frailty group increased,while the body mass in-dex and the proportion of men decreased,and the differences were statistically significant(P＜0.05).The SII and RDW levels in the non-frailty group were lower than those in the frailty group,and the 25(OH)D level was higher than that in the frailty group,and the differences were statistically significant(P＜0.05).Pearson correlation analysis showed that SII and RDW levels were positively correlated with frailty index,and 25(OH)D level was negatively correlated with frailty index in elderly T2DM patients(P＜0.05).Logistic regression analysis showed that female,age ≥ 74.25 years old,SII≥ 938.36,RDW≥ 15.19％,and 25(OH)D≥48.42 nmol/L were independent risk factors for frailty in elderly T2DM patients(P＜0.05).Conclusion The levels of SII,RDW and 25(OH)D in elderly patients with T2DM are related to the frailty index.

Objective To investigate the effect of dorsal repulsive axon guidance protein(Draxin)on the migration of trunk neural crest cells during the early development of embryonic mouse spinal cord.Methods Immunohistochemistry and in situ hybridization were used to detect the expression characteristics of Draxin in early embryonic spinal cord(8 mice each group);In situ hybridization was used to detect the change of migration characteristics of trunk neural crest cells in early embryonic spinal cord of different types of mouse(5 mice each group);in vitro culture method was used to check the effect of Draxin on the migration characteristics of embryonic mouse trunk neural crest cells(16 mice each group).Resultsβ-galactosidase gene Z(LacZ)gene was introduced when Draxin gene was knocked out to produce Draxin gene knockout mice.β-galactosidase staining was used to detect LacZ gene expression in Draxin knockout embryonic mice,and the result showed that Draxin expression was observed in the spinal cord of early embryonic mice since 9.5 days(E9.5).Draxin expression was obvious in the embryonic mice spinal cord in E10.5 period.In situ hybridization was used to detect the expression of Draxin gene in the spinal cord of wild type embryonic mice,and the result further verified the obvious expression of Draxin in the early embryonic mice spinal cord in El0.5 period.Sox10 in situ hybridization was used to detect neural crest cell migration in the spinal cord of embryonic mice in E10.5 period.The result showed that segmental migration of neural crest cells in the early embryonic spinal cord of some Draxin knockout mice was delayed compared with the wild type mice.The effect of Draxin on the migration of wild type early embryonic mice trunk neural crest cells in vitro was tested.The result showed that Draxin reduced the migration distance of neural crest cells in vitro.Conclusion In the early developmental stage of embryonic spinal cord(E9.5-E10.5),neural crest cells migrated exuberant.At the same time,Draxin plays an important inhibitory function in the formation of the specific migration pathways of trunk neural crest cells by promoting neural crest cells migrating away from Draxin expressing regions.

Background: Maturity-onset diabetes of the young (MODY) due to variants of hepatocyte nuclear factor 1-beta (HNF1β) (MODY5) has not been well studied in the Chinese population. This study aimed to estimate its prevalence and evaluate the application of a clinical screening method (Faguer score) in Chinese early-onset diabetes (EOD) patients. Methods: Among 679 EOD patients clinically diagnosed with type 2 diabetes mellitus (age at diagnosis ≤40 years), the exons of HNF1β were sequenced. Functional impact of rare variants was evaluated using a dual-luciferase reporter system. Faguer scores ≥8 prompted multiplex ligation-dependent probe amplification (MLPA) for large deletions. Pathogenicity of HNF1β variants was assessed following the American College of Medical Genetics and Genomics (ACMG) guidelines. Results: Two rare HNF1β missense mutations (E105K and G454R) were identified by sequencing in five patients, showing functional impact in vitro. Another patient was found to have a whole-gene deletion by MLPA in 22 patients with the Faguer score above 8. Following ACMG guidelines, six patients carrying pathogenic or likely pathogenic variant were diagnosed with MODY5. The estimated prevalence of MODY5 in Chinese EOD patients was approximately 0.9% or higher. Conclusion MODY5 is not uncommon in China. The Faguer score is helpful in deciding whether to perform MLPA analysis on patients with negative sequencing results.