A liquid chromatography-tandem mass spectrometry(LC-MS/MS)method was developed for determination of three kinds of β-agonists(Clenbuterol(CL),Ractopamine(RAC)and Salbutamol(SAL))residues in animal liver samples.The liver sample homogenates were extracted with organic solvent,followed by clean-up using the automatic magnetic solid-phase extraction(MSPE),and then analyzed using LC-MS/MS.The results showed that the magnetic mixed-mode cation exchange adsorbent(M-MCX)exhibited 34%higher adsorption capacity than the conventional mixed-mode cation exchange(MCX)column.Furthermore,the clean-up was conducted by using an automatic MSPE device,and 8 samples could be simultaneously treated within 30 min.The limits of detection(LOD)were 0.01-0.1 μg/kg,the average recoveries ranged from 88.2%to 110.5%,and the relative standard deviations(RSDs)were in range of 2.9%-10.3%at three spiked levels for the three kinds of β-agonists.Compared with the traditional SPE technique,the present method had many advantages such as simple operation,rapidity and high efficiency,which was suitable for high-throughput and automatic detection of residues in routine analysis.

Objective To investigate the effect of rutin on pyroptosis of nucleus pulposus cells in rats with intervertebral disc degeneration and its mechanism at cellular level.Methods Rat interdisc nucleus pulposus cells were randomly divided into control group,model group[interleukin-1β(IL-1β)induction],experiment-L group(25 μmol·L-1 rutin+10 ng·mL-1 IL-1β),experiment-M group(50 μmol·L-1 rutin+10ng·mL-1IL-1β),experiment-H group(100 μmol·L-1 rutin+10 ng·mL-1 IL-1 β),experiment-H+SRT1720 agonist group[100 μmol·L-1 rutin+10 ng·mL-1 IL-1 β+1 μmol·L-1 silent information regulator 1(SIRT1)agonist SRT1720].Real-time quantitative polymerase chain reaction(RT-qPCR)assay and Western blot assay were used to detect the expression of related genes and proteins;cell count kit-8(CCK-8)assay and enzyme-linked immunosorbent assay were used to detect the expression of cell viability and inflammatory factors,respectively.Results The cell activities of control group,model group,experimental-H group and experimental-H+SIRT1 agonist group were(100.00±3.56)％,(47.30±2.58)％,(57.33±3.41)％,(66.15±4.85)％,(76.78±2.83)％and(85.84±4.84)％;SIRT1 mRNA expression levels were 1.00±0.12,0.25±0.02,0.39±0.04,0.52±0.03,0.67±0.08 and 0.83±0.09;tumor necrosis factor-α(TNF-α)expression level were(15.03±1.21),(126.50±8.84),(112.60±12.29),(83.71±6.85),(53.88±3.43)and(28.19±4.02)pg·mL-1;the relative expression levels of PTEN-induced putative kinase 1(PINK1)protein were 0.30±0.03,0.26±0.04,0.39±0.04,0.65±0.06,0.89±0.12 and 1.28±0.08;the relative expression levels of gasdermin D-N(GSDMD-N)were 0.38±0.02,1.20±0.08,0.89±0.06,0.69±0.07,0.53±0.05 and 0.40±0.03,respectively.Compared with the model group,the above indexes in the experimental-H group were statistically different(all P＜0.05).There were statistically significant differences in the above indexes between the experimental-H+SIRT1 agonist group and the experimental-H group(all P＜0.05).Conclusion Rutin can induce mitochondrial autophagy by activating SIRT1,inhibit pyroptosis of nucleus pulposus cells and alleviate intervertebral disc degeneration in rats.

Objective To investigate the mechanism of the interaction between Wnt/β-catenin signaling and the epithelial mesenchymal transition(EMT)pathway in mediating sunitinib-resistance in renal cancer cells.Methods The sunitinib-resistant kidney cancer cell lines were constructed by stepwise increase in drug concentrations method with sunitinib,and were divided into resistance group,lv-NC group and lv-Twist group,and human kidney cancer cell lines were selected as normal group.The normal and drug-resistant groups were treated with conventional culture;the lv-NC group was treated with 40 μL of lv-NC lentivirus supernatant containing 2.25 × 108 TU·mL-1 for 72 h,and the lv-Twist group was treated with 50 μL of Twist lentivirus supernatant containing 1.64 × 108 TU·mL-1 for 72 h.The apoptosis ability was detected by flow cytometry;the cell migration ability was detected by cell scratch assay;the cell invasion ability was detected by Transwell assay;and the protein expression levels of Wnt1,β-catenin and Twist were detected by Western blotting assay.Results The apoptosis rates of control,resistant,lv-NC and lv-Twist groups were(17.60±0.59)％,(8.61±0.34)％,(8.60±0.40)％and(3.10±0.34)％;the migration rates were(14.10±0.12)％,(27.64±0.41)％,(14.24±0.45)％and(32.74±2.53)％;the number of invading cells was 27.33±1.15,53.33±1.53,46.00±2.65 and 99.33±2.52;the relative expression levels of Wnt1 protein were 0.10±0.01,0.96±0.06,0.39±0.03 and 3.09±0.31;the relative expression levels of β-catenin protein were 0.39±0.01,1.48±0.16,0.81±0.05 and 1.24±0.14;the relative expression levels of Twist protein were 0.10±0.02,0.91±0.04,0.39±0.03 and 3.09±0.31,respectively.The differences of above indexes were statistically significant between the resistant group and the normal group,and between the lv-Twist group and the lv-NC group(all P＜0.05).Conclusion Twist(EMT related protein)mediates sunitinib resistance in renal cell carcinoma by interacting with Wnt/β-catenin signaling pathway.

Objective To investigate the infection status and changing trend of hepatitis C virus(HCV)infection in hospitalized patients in medical institutions,and provide reference for formulating HCV infection prevention and control strategies.Methods HCV infection surveillance results from cross-sectional survey data reported to China Healthcare-associated Infection(HAI)Surveillance System in 2020 were summarized and analyzed,HCV positive was serum anti-HCV positive or HCV RNA positive,survey result was compared with the survey results from 2003.Results In 2020,1 071 368 inpatients in 1 573 hospitals were surveyed,738 535 of whom underwent HCV test,4 014 patients were infected with HCV,with a detection rate of 68.93％and a HCV positive rate of 0.54％.The positive rate of HCV in male and female patients were 0.60％and 0.48％,respectively,with a statistically sig-nificant difference(x2=47.18,P＜0.001).The HCV positive rate in the 50-＜60 age group was the highest(0.76％),followed by the 40-＜50 age group(0.71％).Difference among all age groups was statistically signifi-cant(x2=696.74,P＜0.001).In 2003,91 113 inpatients were surveyed.35 145 of whom underwent HCV test,resulting in a detection rate of 38.57％;775 patients were infected with HCV,with a positive rate of 2.21％.In 2020,HCV positive rates in hospitals of different scales were 0.46％-0.63％,with the highest in hospital with bed numbers ranging 600-899.Patients'HCV positive rates in hospitals of different scales was statistically signifi-cant(X2=35.34,P＜0.001).In 2020,12 provinces/municipalities had over 10 000 patients underwent HCV-rela-ted test,and HCV positive rates ranged 0.19％-0.81％,with the highest rate from Hainan Province.HCV posi-tive rates in different departments were 0.06％-0.82％,with the lowest positive rate in the department of pedia-trics and the highest in the department of internal medicine.In 2003 and 2020,HCV positive rates in the depart-ment of infectious diseases were the highest,being 7.95％and 3.48％,respectively.Followed by departments of orthopedics(7.72％),gastroenterology(3.77％),nephrology(3.57％)and general intensive care unit(ICU,3.10％)in 2003,as well as departments of gastroenterology(1.35％),nephrology(1.18％),endocrinology(0.91％),and general intensive care unit(ICU,0.79％)in 2020.Conclusion Compared with 2003,HCV positive rate decreased significantly in 2020.HCV infected patients were mainly from the department of infectious diseases,followed by departments of gastroenterology,nephrology and general ICU.HCV infection positive rate varies with gender,age,and region.

Intracellular overexpression of cytoglobin (Cygb) has been shown to reduce extracellular matrix deposition and promote liver fibrosis recovery, but its mechanism is not yet clear. This study constructed and expressed a fusion protein (TAT-Cygb) of cell penetrating peptide TAT and Cygb, to investigate the effect of fusion protein TAT-Cygb on regulating hepatic stellate cells (HSCs) ferroptosis. Cultured human hepatic stellate cells line (LX2) were treated with TAT-Cygb and erastin in vitro, respectively. The effects of ferroptosis phenotype in LX2 cells induced by TAT-Cygb, including cell viability, cell morphology, iron ion (Fe²⁺) content, lipid peroxidation product levels, and antioxidant system indicators, were investigated using trypan blue staining, transmission electron microscopy, Prussian blue staining, and reagent kits detection. After co-treatment with TAT-Cygb and ferrostain-1, the levels of Fe²⁺, reactive oxygen species (ROS), malondialdehyde (MDA), 4-hydroxynonenal (4-HNE), nicotinamide adenine dinucleotide phosphate (NADPH) and glutathione (GSH) were measured by reagent kits. The protein expression levels of alpha smooth actin (α-SMA), collagen I and fibronectin were detected by Western blot, and the protein expression level of epidermal growth factor receptor (EGFR) and desmin relevant to fibrosis were observed by immunofluorescence. The results showed that TAT-Cygb could significantly reduce the viability of LX2 cells and trigger events relevant to ferroptosis, including promoting intracellular Fe²⁺ accumulation, and inducing mitochondrial morphological changes, and intensifying lipid peroxidation products accumulation, and decreasing the level of antioxidant indexes, which played a similar role as erastin; Fer-1 significantly weakened the increase in Fe²⁺, ROS, MDA, 4-HNE levels induced by TAT-Cygb, as well as the decrease in NADPH and GSH levels, while also weakening the TAT-Cygb-induced over-expression levels of α-SMA, collagen I and fibronectin, and TAT-Cygb-induced under-expression levels of EGFR and desmin. This cellular level study indicated that TAT-Cygb can induce ferroptosis of activated HSCs. This study revealed the potential mechanism of TAT-Cygb anti-liver fibrosis, and provided the experimental basis for further research on the molecular mechanism of TAT-Cygb realizing biological function by regulating the ferroptosis pathway.

Objective To develop a deep learning model for automated age estimation based on 3D CT reconstructed images of Han population in western China,and evaluate its feasibility and reliability.Methods The retrospective pelvic CT imaging data of 1 200 samples(600 males and 600 females)aged 20.0 to 80.0 years in western China were collected and reconstructed into 3D virtual bone models.The images of the ischial tuberosity feature region were extracted to create sex-specific and left/right site-specific sample libraries.Using the ResNet34 model,500 samples of different sexes were randomly selected as training and verification set,the remaining samples were used as testing set.Initialization and transfer learning were used to train images that distinguish sex and left/right site.Mean absolute error(MAE)and root mean square error(RMSE)were used as primary indicators to evaluate the model.Results Prediction results varied between sexes,with bilateral models outperformed left/right unilateral ones,and transfer learning models showed superior performance over initial models.In the prediction results of bilateral transfer learning models,the male MAE was 7.74 years and RMSE was 9.73 years,the female MAE was 6.27 years and RMSE was 7.82 years,and the mixed sexes MAE was 6.64 years and RMSE was 8.43 years.Conclusion The skeletal age estimation model,utilizing is-chial tuberosity images of Han population in western China and employing the ResNet34 combined with transfer learning,can effectively estimate adult ischium age.

Objective To describe the current state of research and future research hotspots through a metrological analysis of the literature in the field of forensic anthropological remains identification re-search.Methods The data retrieved and extracted from the Web of Science Core Collection (WoSCC),the core database of the Web of Science information service platform (hereinafter referred to as "WoS"),was used to analyze the trends and topic changes in research on forensic identification of human re-mains from 1991 to 2022.Network visualisation of publication trends,countries (regions),institutions,authors and topics related to the identification of remains in forensic anthropology was analysed using python 3.9.2 and Gephi 0.10.Results A total of 873 papers written in English in the field of forensic anthropological remains identification research were obtained.The journal with the largest number of publications was Forensic Science International (164 articles).The country (region) with the largest number of published papers was China (90 articles).Katholieke Univ Leuven (Netherlands,21 articles) was the institution with the largest number of publications.Topic analysis revealed that the focus of forensic anthropological remains identification research was sex estimation and age estimation,and the most commonly studied remains were teeth.Conclusion The volume of publications in the field of forensic anthropological remains identification research has a distinct phasing.However,the scope of both international and domestic collaborations remains limited.Traditionally,human remains identifica-tion has primarily relied on key areas such as the pelvis,skull,and teeth.Looking ahead,future re-search will likely focus on the more accurate and efficient identification of multiple skeletal remains through the use of machine learning and deep learning techniques.

In infants with severe bronchopulmonary dysplasia(sBPD),severe pulmonary lobar emphysema may occur as a complication,contributing to significant impairment in ventilation.Clinical management of these infants is extremely challenging and some may require lobectomy to improve ventilation.However,prior to the lobectomy,it is very difficult to assess whether the remaining lung parenchyma would be able to sustain adequate ventilation postoperatively.In addition,preoperative planning and perioperative management are also quite challenging in these patients.This paper reports the utility of selective bronchial occlusion in assessing the safety and efficacy of lobectomy in a case of sBPD complicated by severe right upper lobar emphysema.Since infants with sBPD already have poor lung development and significant lung injury,lobectomy should be viewed as a non-traditional therapy and be carried out with extreme caution.Selective bronchial occlusion test can be an effective tool in assessing the risks and benefits of lobectomy in cases with sBPD and lobar emphysema.However,given the technical difficulty,successful application of this technique requires close collaboration of an experienced interdisciplinary team.

Objective To understand the antimicrobial resistance of Escherichia spp.from member units of Hu-nan Province Antimicrobial Resistance Surveillance System from 2012 to 2021.Methods According to the technical scheme of China Antimicrobial Resistance Surveillance System(CARSS),data about Escherichia spp.and the anti-microbial susceptibility testing results reported from member units of Hunan Province Antimicrobial Resistance Sur-veillance System were analyzed by WHONET 5.6 software.Results From 2012 to 2021,a total of 476 351 clini-cally isolated Escherichia spp.were collected,475 520 of which were Escherichia coli,accounting for 99.8％;92.6％were isolated from inpatients;39.3％were isolated from urine specimens.Over the past 10 years,the proportion of Escherichia spp.in total detected pathogens remained relatively stable,ranging 20％-23％,the lowest rate was 18.7％in 2012,and the highest rate was 22.9％in 2015.In the past 10 years,the resistance rates of Escherichia spp.to ampicillin,ceftriaxone,cefotaxime and ampicillin/sulbactam were＞80％,＞47％,＞45％,and＞39％,respectively;resistance rates to piperacillin/tazobactam,cefoperazone/sulbactam,and nitrofurantoin were all＜8％,to tigecycline,amikacin,imipenem,and meropenem(except in 2012)were all＜5％.Resistance of Escherichia spp.to 22 commonly clinically used antimicrobial agents fluctuated,but overall trend decreased year by year.The resistance rates of Escherichia spp.from patients in the intensive care unit(ICU),non-ICU patients,outpatients,and emergency patients to 22 clinically commonly used antimicrobial agents were compared among different depart-ments,and the differences were statistically significant(all P＜0.05).The resistance rates of Escherichia spp.iso-lated from ICU and non-ICU patients were compared,and except for tigecycline,the resistance rates to the other 21 antimicrobial agents were statistically different(all P＜0.05).The resistance rates of Escherichia spp.isolated from patients to commonly clinically used antimicrobial agents were statistically different among patients of different age groups(all P＜0.05).Conclusion Escherichia spp.isolated from patients in different years,departments,specimens,and ages have different resistance to commonly used antimicrobial agents.It is necessary to continue to strengthen the surveillance on bacterial resistance,so as to guide the rational choice of antimicrobial agents.

Candida auris(C.auris)is a fungus with multidrug resistance.Current genomic studies indicate that C.auris shares similarities in genome size and evolution with other resistant yeast species.There are differences in genome size and structural variations among different geographical clades of C.auris,which may be related to differences in phenotype and drug resistance.C.auris exhibits resistance to multiple antifungal agents,including triazole,amphotericin B,and echinocandins.Genomic studies have found that resistance of C.auris may be related to factors such as membrane transport protein and mutation in the ergosterol pathway,its resistance can change fur-ther during evolution.In this paper,the relevant studies are reviewed,with a view to understanding the mechanism of drug resistance of C.auris,and providing important basis for formulating prevention and control strategies as well as treatment programs of C.auris.