ObjectiveTaking the cuproptosis/oxidative stress pathway as the entry point， this study investigated the effect and mechanism of Liangyi Paste on hepatic lipid deposition in naturally aged mice fed with a high-fat diet. MethodsAfter adaptive feeding， 80 ten-week-old male C57BL/6 mice were used. Thirty of them were randomly divided into three groups （10 mice per group）： The 12-month-old control group （12MCON）， the 15-month-old control group （15MCON）， and the 15-month-old group with a high-fat diet （15MHFD）. The 12MCON and 15MCON groups were continuously fed a standard diet， while the 15MHFD group started receiving a high-fat diet at 12 months of age. Tissue samples were collected at the corresponding time points for each group. The remaining 50 mice were randomly divided into five groups （10 mice per group）： the 20-month-old control group （20MCON）， the model group， and the low-， medium-， and high-dose Liangyi Paste groups （2.91 ， 5.82 ， 11.64 g·kg^-1·d^-1， respectively）. The 20MCON group was continuously fed a standard diet， while the other groups started receiving a high-fat diet at 15 months of age. At 18 months of age， the Liangyi Paste groups were administered the corresponding doses of Liangyi Paste by gavage， while the 20MCON and model groups were given an equal volume of saline by gavage. After 8 weeks of continuous gavage （when the mice reached 20 months of age）， tissue samples were collected. Hepatic TG levels were measured using assay kits； liver histology and lipid deposition were observed via hematoxylin-eosin （HE） and oil red O staining； reactive oxygen species （ROS） were detected by enzyme-linked immunosorbent assay （ELISA）； Cu²⁺， superoxide dismutase （SOD）， and malondialdehyde （MDA） levels were measured by colorimetry； mRNA and protein expression of genes related to cuproptosis and oxidative stress pathways were analyzed by Real-time polymerase chain reaction（Real-time PCR） and Wes automated protein expression system. ResultsCompared with 12MCON， the 15MCON group showed significantly increased hepatic TG， Cu²⁺， ROS， and MDA levels （P<0.01）， decreased SOD （P<0.01）， hepatocyte swelling， and disordered arrangement. The mRNA and protein levels of ferredoxin 1 （FDX1）， dihydrolipoamide S-acetyltransferase （DLAT）， heat shock protein 70 （HSP70）， dihydrolipoamide dehydrogenase （DLD）， pyruvate dehydrogenase E1 subunit-β （PDHB）， nuclear factor erythroid 2-related factor 2 （Nrf2）， and peroxisome proliferator-activated receptor γ （PPARγ） were significantly elevated （P<0.05， P<0.01）. Compared with 15MCON group， the 15MHFD and 20MCON groups exhibited further increases in TG， Cu²⁺， ROS， and MDA （P<0.01）， reduced SOD （P<0.01）， and aggravated hepatocyte swelling and disorder. There were increased lipid droplets with mild vacuolization in the 15MHFD group， and no significant lipid deposition was observed in the 20MCON group. FDX1， DLAT， HSP70， DLD， PDHB， Nrf2， and PPARγ mRNA and protein levels were significantly increased （P<0.05， P<0.01）. Compared with 20MCON group， the model group demonstrated markedly elevated TG， Cu²⁺， ROS， and MDA （P<0.01）， reduced SOD （P<0.01）， severe hepatic steatosis， and upregulated expression of FDX1， DLAT， HSP70， DLD， PDHB， Nrf2， and PPARγ mRNA and proteins （P<0.05， P<0.01）. All abnormalities were significantly reversed after Liangyi Paste treatment. ConclusionLiangyi paste can ameliorate hepatic lipid deposition in naturally aged mice with a high-fat diet by modulating the cuproptosis/oxidative stress pathway.

BACKGROUND:Cell-free therapy is a research hotspot in the field of medical cosmetic anti-aging.It is still unknown for paracellular secretion of human umbilical cord mesenchymal stem cell-derived small extracellular vesicles loaded with the antiaging drug α-ketoglutaric acid to delay skin aging.OBJECTIVE:To investigate the effect of the anti-aging agent α-ketoglutarate engineered human umbilical cord mesenchymal stem cell-derived small extracellular vesicles in a D-galactose-induced model of dermal fibroblast senescence.METHODS:(1)Biological characteristics of primary human umbilical cord mesenchymal stem cells were identified by osteogenic-lipogenic differentiation staining and flow cytometry.(2)The small extracellular vesicles derived from human umbilical cord mesenchymal stem cell were obtained by using differential-ultracentrifugation.α-Ketoglutarate-engineered human umbilical cord mesenchymal stem cell-small extracellular vesicles were constructed by electroporation,and biologically characterized by transmission electron microscopy and nanoparticle tracking analyzer,while the encapsulation rate was assessed using high-performance liquid chromatography.(3)The effect of α-ketoglutarate on the proliferative capacity of dermal fibroblasts was assessed by CCK-8 and Edu cell proliferation assay kits.(4)The effect of α-ketoglutarate-engineered human umbilical cord mesenchymal stem cell-small extracellular vesicles on delaying the senescence of dermal fibroblasts was evaluated by reactive oxygen species detection kit,western blot assay,and cellular immunofluorescence.RESULTS AND CONCLUSION:(1)The obtained human umbilical cord mesenchymal stem cell and human umbilical cord mesenchymal stem cell-small extracellular vesicles were biologically compatible.(2)There was no toxic effect on dermal fibroblasts when α-ketoglutarate was used in the concentration range of 0.5-8 mmol/L.(3)D-gal induced senescence in dermal fibroblasts,while α-ketoglutarate-engineered human umbilical cord mesenchymal stem cell-small extracellular vesicles treatment reduced the level of oxidative stress,DNA damage,and collagen loss,which was further verified that α-ketoglutarate-engineered human umbilical cord mesenchymal stem cell-small extracellular vesicles could effectively slow down the skin aging process.

BACKGROUND:Cell-free therapy is a research hotspot in the field of medical cosmetic anti-aging.It is still unknown for paracellular secretion of human umbilical cord mesenchymal stem cell-derived small extracellular vesicles loaded with the antiaging drug α-ketoglutaric acid to delay skin aging.OBJECTIVE:To investigate the effect of the anti-aging agent α-ketoglutarate engineered human umbilical cord mesenchymal stem cell-derived small extracellular vesicles in a D-galactose-induced model of dermal fibroblast senescence.METHODS:(1)Biological characteristics of primary human umbilical cord mesenchymal stem cells were identified by osteogenic-lipogenic differentiation staining and flow cytometry.(2)The small extracellular vesicles derived from human umbilical cord mesenchymal stem cell were obtained by using differential-ultracentrifugation.α-Ketoglutarate-engineered human umbilical cord mesenchymal stem cell-small extracellular vesicles were constructed by electroporation,and biologically characterized by transmission electron microscopy and nanoparticle tracking analyzer,while the encapsulation rate was assessed using high-performance liquid chromatography.(3)The effect of α-ketoglutarate on the proliferative capacity of dermal fibroblasts was assessed by CCK-8 and Edu cell proliferation assay kits.(4)The effect of α-ketoglutarate-engineered human umbilical cord mesenchymal stem cell-small extracellular vesicles on delaying the senescence of dermal fibroblasts was evaluated by reactive oxygen species detection kit,western blot assay,and cellular immunofluorescence.RESULTS AND CONCLUSION:(1)The obtained human umbilical cord mesenchymal stem cell and human umbilical cord mesenchymal stem cell-small extracellular vesicles were biologically compatible.(2)There was no toxic effect on dermal fibroblasts when α-ketoglutarate was used in the concentration range of 0.5-8 mmol/L.(3)D-gal induced senescence in dermal fibroblasts,while α-ketoglutarate-engineered human umbilical cord mesenchymal stem cell-small extracellular vesicles treatment reduced the level of oxidative stress,DNA damage,and collagen loss,which was further verified that α-ketoglutarate-engineered human umbilical cord mesenchymal stem cell-small extracellular vesicles could effectively slow down the skin aging process.

In recent years,natural antimicrobial biomaterials have received widespread attention due to their rich sources,broad anti-microbial spectrum,high antimicrobial activity,good biocompatibility,etc.They play an important role in caries prevention and treat-ment by inhibiting the formation of biofilm and removing the formed biofilm.In this paper,we summarize the inhibitory effects of differ-ent natural antimicrobial biomaterials on Streptococcus mutans,the main caries-causing organism,and its biofilm,with a view to provi-ding theoretical references for caries prevention and treatment.

ObjectiveTo understand the epidemic characteristics of influenza in Beijing from 2023 to 2024, and to provide a scientific basis for the prevention and control of influenza. MethodsData on influenza-like illness (ILI) from secondary level and above hospitals, etiology surveillance data, and influenza clusters outbreaks data from 2023‒2024 were used to analyze the epidemic trend and pathogenic characteristics of influenza. Furthermore, an influenza comprehensive index was used to categorize the epidemic intensity at the severity level. ResultsA total of 2 065 857 ILI cases were reported in 2023‒2024 epidemic season, and the percentage of ILI was 3.67%. The age group of 5‒14 years accounted for the highest proportion of ILI (30.48%). A total of 41 766 throat swabs from ILI were detected, with a positive rate of 17.28%.A (H3N2) (51.86%) and B Victoria (41.93%) were the most prevalent subtypes of influenza virus. Clustered influenza outbreaks occurred mainly in primary schools (57.78%) and middle schools (35.55%), mainly caused by the influenza A (H3N2) subtype (85.93%). According to the influenza comprehensive index (I), the period of influenza activity and above (I>0.5) lasted for a total of 37 weeks, accounting for 71.15% of the entire influenza season. ConclusionCompared with previous years, the epidemic level of influenza in Beijing was increased in 2023‒2024, and the peak time became earlier. The comprehensive index method can objectively evaluate the level of influenza epidemic and provide suggestions for the future prevention and control of influenza in Beijing.

Objective:To investigate the intervention effect and active mechanism of traditional Chinese medicine Marsdenia tenacis-sima on colitis-associated colorectal cancer(CAC).Methods:High-resolution mass spectrometry and a precise mass number data-base were used to identify the chemical components of Marsdenia tenacissima,and network pharmacology methods were used to obtain the main active components and targets of Marsdenia tenacissima in the treatment of colorectal cancer(CRC)and analyze its mecha-nism of action against CRC.A mouse model of CAC was established,and then Marsdenia tenacissima was analyzed in terms of its effect on disease activity index(DAI),body mass,colorectal morphology,and serum levels of tumor necrosis factor-α(TNF-α)and interleu-kin-1β(IL-1β)in CAC mice.Furthermore,16S rDNA sequencing technique was used to assess the impact of Marsdenia tenacissima on the diversity of intestinal microbiota in mice.Results:A total of 50 components were identified by mass spectrometry,and 37 active components were identified by network pharmacology.Furthermore,313 targets related to CRC were identified.A total of 958 GO terms and 173 KEGG terms were obtained,among which inflammation,oxidative stress,and cancer pathways showed a relatively high de-gree of association with Marsdenia tenacissima.After modeling,the mice in each group had the manifestations such as diarrhea,bloody stool,prolapse of anus,and weight loss,and the model group had a significant increase in DAI score after each DSS cycle,while the Marsdenia tenacissima group had a significant reduction in DAI score of the model mice after the first,second,and third cycles(P=0.007,P<0.001,and P<0.001).In addition,compared with the model group,the Marsdenia tenacissima group had a significant in-crease in colorectal length,alleviation of intestinal swelling,a relatively intact structure of epithelial cells,a reduction in inflammatory cell infiltration,a significant increase in goblet cells,a reduction in atypical hyperplasia,and significant reductions in the levels of TNF-α and IL-6 in CAC mice(P<0.001 and 0.002).The Marsdenia tenacissima group had significant reductions in the abundance and diversity of intestinal microbiota,and compared with the model group,the Marsdenia tenacissima group had a significant reduction in the relative abundance of Bacteroides and significant increases in the relative abundances of Muribaculaceae and Clostridia-UCG-014(P=0.002 and 0.009).Conclusion:Marsdenia tenacissima exerts an intervention effect on colorectal cancer in CAC mice,possibly by regulating the pathways associated with inflammation and cancer and altering the diversity of intestinal microbiota.

Objective:To identify the risk factors associated with conversion to cesarean delivery in parturients receiving epidural labor analgesia and to construct a predictive model.Methods:In this prospective cohort study, 718 parturients with a singleton fetus at 37-42 weeks of gestation age, aged 22-45 yr, of American Society of Anesthesiologists Physical Status classification Ⅱ, who received epidural labor analgesia at the Women′s Hospital affiliated to Zhejiang University School of Medicine between November 2022 and August 2023, were included and divided into 2 groups based on the final mode of delivery: cesarean delivery group (CD group) and vaginal delivery group (VD group). Data collected included maternal age, height, body mass index at labor onset, gestational age, pregnancy complications, mode of conception, parity, premature rupture of membranes, mode of labor onset and details of labor analgesia. Risk factors for conversion to cesarean delivery were analyzed using multivariate logistic regression analysis, and a prediction model was developed. The receiver operating characteristic curve was ploted, and the area under the receiver operating characteristic curve was calculated to evaluate the accuracy of the predictive model.Results:A total of 682 parturients were included in the final statistical analysis, with 103 in CD group and 579 in VD group. Multivariate logistic regression analysis showed that maternal age≥35 yr, height<160 cm, body mass index at labor onset ≥28 kg/m 2, nulliparous parturients, the use of Foley balloon catheter or dinoprostone suppositories for cervical ripening were risk factors for cesarean delivery, and cervical dilation during analgesia was a protective factor. The area under the receiver operating characteristic curve for the predictive model constructed based on the aforementioned influencing factors yielded predicting conversion to cesarean delivery due to failed vaginal trial of labor was 0.791 (95% confidence interval 0.744-0.837, P<0.001), with a sensitivity of 73.6% and a specificity of 70.9%. Conclusions:Nulliparous parturients, maternal age≥35 yr, height<160 cm, body mass index at labor onset≥28 kg/m 2, and the use of Foley balloon catheter or dinoprostone suppositories for cervical ripening are risk factors for cesarean delivery, and cervical dilation during analgesia is a protective factor. The predictive model established based on these influencing factors has high predictive performance.

Objective:To present a set of clinically representative quality assurance (QA) test cases for stereotactic radiosurgery (SRT) plans of multiple intracranial metastases, in order to assess the plan quality and machine execution capabilities.Methods:Based on the clinical characteristics of multiple brain metastases, four groups of test cases with three target volumes (TVs), six TVs, nine TVs, and TVs near organs at risk (OARs) were designed. For these cases, SRT plans were developed, and plan quality was assessed using metrics including the Radiation Therapy Oncology Group conformality index (RTOG CI), gradient index (GI), homogeneity index (HI), and the volume of normal brain tissue receiving a dose of 24 Gy ( V24 Gy), which was defined as the volume enclosed by the 24 Gy isodose line around the Brain-PTV ( V24 Gy of Brain-PTV). Verification plans were generated for each test case, including the verification of point doses, planar doses (PD), and SRS MapCHECK (SMC) semiconductor matrix planar doses. Compared with the calculated result of the treatment planning system (TPS), the criteria for the γ analysis of planar doses were set at 1 mm/2% and 2 mm/2%. Results:For the four groups of test cases, the mean CI, GI, HI, and V24 Gy of Brain-PTV were 1.04±0.03, 3.79±0.40, 0.73±0.01 and (7.46±3.80) cm 3, respectively. The mean deviations of the point doses were 0.88%±0.98%, 1.47%±0.79%, 1.52%± 0.76%, and 1.17% ± 0.38%, respectively. The mean γ passing rates of the single fields for PDs were greater than 98% at 2 mm/2% and exceeding 96% at 1 mm/2%, and the mean γ pass rates of the SMC semiconductor matrix for PDs were 97.75% ± 2.31% and 99.33% ± 0.62%, at 1 mm/2% and 2 mm/2% respectively. Conclusions:The proposed QA test cases for SRT of multiple intracranial metastases allow for the effective assessments of the plan quality and machine execution capabilities and, thus, can assist various centers in clinical applications.

Objective:To analyze the changes in the prevalence characteristics of multimorbidity among adolescents aged 13-18 in Inner Mongolia Autonomous Region from 2019 to 2022 and to explore the association between multimorbidity and moderate to high-intensity physical activity among them.Methods:A stratified random cluster sampling method was used to select students aged 13-18 in Inner Mongolia Autonomous Region every September from 2019 to 2022. Physical examinations, demographic characteristics, and depression-related surveys were conducted to analyze the multimorbidity of overweight, obesity, high blood pressure, myopia, spinal curvature abnormality, and depression. A logistic regression model was used to analyze the association between multimorbidity and moderate to high-intensity physical activity.Results:From 2019 to 2022, 70 972, 62 923, 80 254, and 78 288 study subjects were included, with the rates of multimorbidity being 56.4%, 55.4%, 57.2%, and 55.8%, respectively. The rates of multimorbidity remained relatively stable from 2019 to 2022 ( χ2=0.06, P=0.950). The incidence of multimorbidity among girls was significantly higher than that among boys ( P<0.001). The incidence of multimorbidity among urban students was significantly higher than that among suburban students ( P<0.001). The incidence of multimorbidity among high school students was higher than that among middle school students ( P<0.001). The top three multimorbidity combinations were myopia and overweight/obesity (26.4%), myopia and high blood pressure (24.4%), and myopia and depression (19.8%), while the least common combination was depression and spinal curvature abnormality (1.1%). The multimorbidity patterns showed no significant differences between years ( χ2=0.03, P=0.999). The multimorbidity status was significantly associated with the status of meeting the standard of moderate to high-intensity physical activity ( OR=0.83, 95% CI: 0.80-0.86). The association was stronger in boys ( OR=0.77, 95% CI: 0.73-0.81) compared with girls ( OR=0.90, 95% CI: 0.85-0.96), with a significant interaction term ( P<0.001). Conclusion:From 2019 to 2022, the incidence of multimorbidity among adolescents aged 13 to 18 in Inner Mongolia Autonomous Region is relatively high, mainly due to the co-occurrence of myopia and other health problems. Adequate physical activity is an important factor in reducing multimorbidity.

Metagenomic next-generation sequencing, as an unbiased detection technology, demonstrates higher diagnostic efficacy than traditional methods. Gut microorganisms are important flora for safeguarding health and have become a hot research topic. Modeling and analyzing the genomic data of intestinal flora using machine learning is very important in disease prediction and diagnosis. This paper briefly introduces the characteristics of metagenomic next-generation sequencing, key algorithms and evaluation indexes of machine learning, outlines the main steps of combining machine learning with metagenomic next-generation sequencing, and summarizes the application of the combination of machine learning and metagenomic next-generation sequencing technology in the study of intestinal flora, which will provide a more accurate method for diagnosis and prediction of the related diseases, and give more ideas for the future research and clinical practice.