Objective To establish a method for determining the concentration of contezolid in human cerebro spinal fluid(CSF)using ultra high performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS).Methods Linezolid as the internal standard(IS)and acetonitrile as the protein precipitant.Waters ACQUITY UPLC? BEH C18(2.1 mm × 50.0 mm,1.7 μm)chromatographic column was used for separation,with a mobile phase of 0.1％formic acid aqueous solution-0.1％formic acid acetonitrile solution,gradient elution method,flow rate was 0.4 mL·min-1,column temperature was 40 ℃,automatic sampler temperature was 10 ℃,the analysis time was 4 minutes.Electrospray ion source,positive ion mode,and multi-reaction monitoring scanning mode were used.The monitoring and analysis ion pairs for contezolid were m/z 409.15→269.14,and the monitoring ion pairs for linezolid were m/z 338.14 → 195.10.The specificity,standard curve and lower limit of quantification(LLOQ),precision and recovery rate,matrix effect,residual effect,dilution effect and stability of the method were investigated.Results The endogenous substances in CSF do not interfere with the determination of the analyte contezolid and the internal standard linezolid,and the method has good specificity.Satisfactory linearity was observed within the concentration range of 20-5 000 ng·mL-1 for contezolid in CSF,the calibration curve was y=8.97 × 10-4x+1.95 × 10-2(r=0.999 1),and the LLOQ was 20 ng·mL-1.Precision of the intra-batch and inter-batch relative standard deviation(RSD)＜15％,and the extraction recovery were 90.96％-98.71％.The average normalized matrix effect factor of the quality control CSF sample were 94.39％-100.25％.The RSD of dilution effect＜15％.The CSF samples of contezolid were stored at room temperature,in the automatic sampler for 72 hours,-20 ℃ and-80 ℃ for 90 days,and subjected to repeated freezing and thawing three times,were stable with all of which the RSD＜10％.Conclusion This method is high sensitivity,rapid,simple and accurate,which is very suitable for the therapeutic drug monitoring of contezolid in human CSF.

Aim To establish and evaluate an alternative meth-od for detecting skin sensitization of cosmetics based on local lymph node assay using bromodeoxyuridine(BrdU)with flow cytometry(FCM).Methods(1)25％hexyl cinnamic alde-hyde(HCA)was chosen as a positive control with an acetone:olive oil(4∶1,V/V,AOO)mixture as a vehicle control for the experiment.The dorsal sides of both ears of mice were treated with test solutions on day 1,day 2,and day 3.Brdu solution was injected inter-peritoneally on day 5.On day 6,the bilateral ears and mandibular lymph nodes were excised,and the number of Brdu positive cells was measured by flow cytometry.The stim-ulation index(SI)was calculated to identify whether it was ≥3,in order to establish the method of LLNA:Brdu-FCM.(2)BrdU-FCM test was conducted using a blind method with the fif-teen reference substances listed in OECD TG429 whose skin sensitization potentials were known.The test substances were dissolved in AOO,N,N-dimethylformamide(DMF)or dimeth-yl sulfoxide(DMSO)at three different concentrations.Tests were performed the same as above.SI and EC2.7 were calculat-ed to evaluate whether the test substance was categorized as a skin sensitizer.The reliability and accuracy of the method were validated by comparing the classification of test substances with that in OECD TG429.Results The SI for 25％HCA was 3.9,showing positive in the skin sensitization test.It demonstrated that the LLNA:Brdu-FCM test method was properly implemen-ted.Nine test substances(2,4-dinitrochlorobenzene,4-pheny-lenediamine,cobalt chloride,2-mercaptobenzothiazole,hexyl-cinnamaldehyde,eugenol,phenyl benzoate,cinnamic alcohol,imidazolidinyl urea)were positive,and six test substances(methyl methacrylate,chlorobenzene,isopropanol,lactic acid,methyl salicylate,salicylic acid)were negative.The method was evaluated with sensitivity of 90％,specificity of 100％,positive prediction rate of 100％,negative prediction rate of 83％,false positive rate of 0％,false negative rate of 17％and accuracy of 93％.The LLNA:BrdU-FCM assay could correctly categorize the test substances that were skin sensitizers or non-sensitizers.Conclusion The LLNA:BrdU-FCM assay appears to be a relia-ble predictor of skin sensitization protential of chemicals,and it is expected to an alternative method for identifying skin sensitization as a supplementary in safety evaluation of cosmetic ingredient.

Aim To explore the mechanism of the syn-ergistic effect of the ferroptosis inducer erastin com-bined with shikonin in promoting ferroptosis in human hypertrophic scar fibroblasts(HSFBs).Methods Hypertrophic scar tissues provided by the General Hos-pital of Ningxia Medical University were collected,and HSFBs were extracted.HSFBs were identified by HE staining and immunofluorescence.The inhibitory rates of Era and SHK on HSFBs at different concentrations were detected by CCK-8 assay,and the IC50 value was calculated.CompuSyn software was used to calculate the co-use index(CI).Control group,Erastin(Era)group,shikonin(SHK)group and Era+SHK group were set up,and the number and morphological chan-ges of cells were observed after 24 hours of interven-tion.The ability of cell migration and invasion was de-tected by scratch test and Transwell test.The changes of malondialdehyde(MDA),total iron ion and reactive oxygen species(ROS)were detected by corresponding biochemical kits.The expressions of collagen I,α-SMA and GOT1,SLC7A11,GPX4 and FTH1 were detected by Western blot.Results The IC50 value of Era and SHK of primary HSFBs was 2.22 μmol·L-1 and 3.94μmol·L-1 respectively,which was used as the single drug concentration for subsequent experiments.The CompuSyn software was employed to calculate the CI value when the two drugs were used in combination,and the concentrations corresponding to CI=0.39597(Era:1.2 μmol·L-1+SHK:1.5 μmol·L-1)were selected as subsequent combination concentrations(Because when CI was equal to 0.395 97,the concen-tration of each drug was lower than the concentration of single drug,and the inhibition rate of combined drug was greater than 50％).Compared with the monother-apy group,the number of HSFBs in the SHK+Era group was significantly reduced,cell membrane showed breakage and vesiculation,cell wrinkling became smal-ler,and cytoplasm was concentrated.The migration and invasion ability of HSFBs in the SHK+Era group were obviously weakened(P＜0.05),and the expres-sion of fibrosis-related proteins collagen Ⅰ and α-SMA was reduced(P＜0.05);the contents of MDA,total i-ron ions,and ROS in HSFBs of the SHK+Era group increased(P＜0.05),and the protein expression lev-els of SLC7A11,GOT1,GPX4,and FTH1 further de-creased(P＜0.05).Conclusions Erastin in combi-nation with shikonin can synergistically inhibit the pro-liferation,migration and fibrosis levels of HSFBs.The mechanism may be that erastin enhances the inhibition of shikotin on GOT1,increases the levels of cellular i-ron ions,ROS,and lipid peroxides,thereby promoting ferroptosis in HSFBs.

Objective To explore the application effect of immersive virtual reality(IVR)technology and different viewing content in pediatric patients with general anesthesia during elective surgery,and to provide references for clinical implementation.Methods A total of 180 pediatric patients who underwent elective surgery under general anesthesia in a tertiary hospital in Qingdao from February to October 2023 were selected as study population using convenient sampling method.According to the operation time,60 pediatric patients who underwent surgery from May to July 2023 were included in the immersive panoramic surgical education group,and they could watch the panoramic surgical education video immersively on the basis of routine care.A total of 60 pediatric patients who underwent surgery from August to October 2023 were included in the immersive animation group to watch cartoons immersively on the basis of routine care.A total of 60 pediatric patients who underwent surgery from February to April 2023 were included in the control group and routine preoperative care was implemented.The preoperative anxiety levels,anesthesia induction compliance,and incidence of emergence agitation were compared in the 3 groups by the modified Yale Preoperative Anxiety Scale-Short Form(mYPAS-SF),anesthesia induction cooperation grade,and the Pediatric Anesthesia Emergence Delirium scale(PAED).Results There were statistically significant differences in preoperative anxiety level,anesthesia induced compliance and incidence of emergence agitation during awakening between the 3 groups(P＜0.001).Among them,the preoperative anxiety level of the immersive panoramic surgical education group and the immersive animation group was lower than that of the control group,and the difference was statistically significant(P＜0.001).The anesthesia induced compliance degree of the immersive panoramic surgical education group and the immersive animation group was better than that of the control group,and the difference was statistically significant(P＜0.017),and the incidence of emergence agitation in the immersive panoramic surgical education group was lower than that of the control group,and the difference was statistically significant(P=0.004).Conclusion The use of IVR technology to watch panoramic surgical education videos and cartoons can help reduce the preoperative anxiety level and improve anesthesia induction cooperation degree of pediatric patients with general anesthesia during elective surgery,but the intervention effect of panoramic surgical education videos is better in improving the emergence agitation.

Objective To establish a method for determining the concentration of contezolid in human cerebro spinal fluid(CSF)using ultra high performance liquid chromatography tandem mass spectrometry(UPLC-MS/MS).Methods Linezolid as the internal standard(IS)and acetonitrile as the protein precipitant.Waters ACQUITY UPLC? BEH C18(2.1 mm × 50.0 mm,1.7 μm)chromatographic column was used for separation,with a mobile phase of 0.1％formic acid aqueous solution-0.1％formic acid acetonitrile solution,gradient elution method,flow rate was 0.4 mL·min-1,column temperature was 40 ℃,automatic sampler temperature was 10 ℃,the analysis time was 4 minutes.Electrospray ion source,positive ion mode,and multi-reaction monitoring scanning mode were used.The monitoring and analysis ion pairs for contezolid were m/z 409.15→269.14,and the monitoring ion pairs for linezolid were m/z 338.14 → 195.10.The specificity,standard curve and lower limit of quantification(LLOQ),precision and recovery rate,matrix effect,residual effect,dilution effect and stability of the method were investigated.Results The endogenous substances in CSF do not interfere with the determination of the analyte contezolid and the internal standard linezolid,and the method has good specificity.Satisfactory linearity was observed within the concentration range of 20-5 000 ng·mL-1 for contezolid in CSF,the calibration curve was y=8.97 × 10-4x+1.95 × 10-2(r=0.999 1),and the LLOQ was 20 ng·mL-1.Precision of the intra-batch and inter-batch relative standard deviation(RSD)＜15％,and the extraction recovery were 90.96％-98.71％.The average normalized matrix effect factor of the quality control CSF sample were 94.39％-100.25％.The RSD of dilution effect＜15％.The CSF samples of contezolid were stored at room temperature,in the automatic sampler for 72 hours,-20 ℃ and-80 ℃ for 90 days,and subjected to repeated freezing and thawing three times,were stable with all of which the RSD＜10％.Conclusion This method is high sensitivity,rapid,simple and accurate,which is very suitable for the therapeutic drug monitoring of contezolid in human CSF.

Aim To establish and evaluate an alternative meth-od for detecting skin sensitization of cosmetics based on local lymph node assay using bromodeoxyuridine(BrdU)with flow cytometry(FCM).Methods(1)25％hexyl cinnamic alde-hyde(HCA)was chosen as a positive control with an acetone:olive oil(4∶1,V/V,AOO)mixture as a vehicle control for the experiment.The dorsal sides of both ears of mice were treated with test solutions on day 1,day 2,and day 3.Brdu solution was injected inter-peritoneally on day 5.On day 6,the bilateral ears and mandibular lymph nodes were excised,and the number of Brdu positive cells was measured by flow cytometry.The stim-ulation index(SI)was calculated to identify whether it was ≥3,in order to establish the method of LLNA:Brdu-FCM.(2)BrdU-FCM test was conducted using a blind method with the fif-teen reference substances listed in OECD TG429 whose skin sensitization potentials were known.The test substances were dissolved in AOO,N,N-dimethylformamide(DMF)or dimeth-yl sulfoxide(DMSO)at three different concentrations.Tests were performed the same as above.SI and EC2.7 were calculat-ed to evaluate whether the test substance was categorized as a skin sensitizer.The reliability and accuracy of the method were validated by comparing the classification of test substances with that in OECD TG429.Results The SI for 25％HCA was 3.9,showing positive in the skin sensitization test.It demonstrated that the LLNA:Brdu-FCM test method was properly implemen-ted.Nine test substances(2,4-dinitrochlorobenzene,4-pheny-lenediamine,cobalt chloride,2-mercaptobenzothiazole,hexyl-cinnamaldehyde,eugenol,phenyl benzoate,cinnamic alcohol,imidazolidinyl urea)were positive,and six test substances(methyl methacrylate,chlorobenzene,isopropanol,lactic acid,methyl salicylate,salicylic acid)were negative.The method was evaluated with sensitivity of 90％,specificity of 100％,positive prediction rate of 100％,negative prediction rate of 83％,false positive rate of 0％,false negative rate of 17％and accuracy of 93％.The LLNA:BrdU-FCM assay could correctly categorize the test substances that were skin sensitizers or non-sensitizers.Conclusion The LLNA:BrdU-FCM assay appears to be a relia-ble predictor of skin sensitization protential of chemicals,and it is expected to an alternative method for identifying skin sensitization as a supplementary in safety evaluation of cosmetic ingredient.

Objective To explore the application effect of immersive virtual reality(IVR)technology and different viewing content in pediatric patients with general anesthesia during elective surgery,and to provide references for clinical implementation.Methods A total of 180 pediatric patients who underwent elective surgery under general anesthesia in a tertiary hospital in Qingdao from February to October 2023 were selected as study population using convenient sampling method.According to the operation time,60 pediatric patients who underwent surgery from May to July 2023 were included in the immersive panoramic surgical education group,and they could watch the panoramic surgical education video immersively on the basis of routine care.A total of 60 pediatric patients who underwent surgery from August to October 2023 were included in the immersive animation group to watch cartoons immersively on the basis of routine care.A total of 60 pediatric patients who underwent surgery from February to April 2023 were included in the control group and routine preoperative care was implemented.The preoperative anxiety levels,anesthesia induction compliance,and incidence of emergence agitation were compared in the 3 groups by the modified Yale Preoperative Anxiety Scale-Short Form(mYPAS-SF),anesthesia induction cooperation grade,and the Pediatric Anesthesia Emergence Delirium scale(PAED).Results There were statistically significant differences in preoperative anxiety level,anesthesia induced compliance and incidence of emergence agitation during awakening between the 3 groups(P＜0.001).Among them,the preoperative anxiety level of the immersive panoramic surgical education group and the immersive animation group was lower than that of the control group,and the difference was statistically significant(P＜0.001).The anesthesia induced compliance degree of the immersive panoramic surgical education group and the immersive animation group was better than that of the control group,and the difference was statistically significant(P＜0.017),and the incidence of emergence agitation in the immersive panoramic surgical education group was lower than that of the control group,and the difference was statistically significant(P=0.004).Conclusion The use of IVR technology to watch panoramic surgical education videos and cartoons can help reduce the preoperative anxiety level and improve anesthesia induction cooperation degree of pediatric patients with general anesthesia during elective surgery,but the intervention effect of panoramic surgical education videos is better in improving the emergence agitation.

This study analyzed the burden and trends in enteric infections in China from 1990 to 2021 from a One Health perspec-tive.Data on mortality associated with enteric infections were extracted from the 2021 Global Burden of Disease(GBD)database.The analysis focused on assessing the mortality rates of enteric infectious diseases attributed to various etiologies and risk factors,along with the age and sex distribution,from 1990 to 2021.Average annual percentage change(AAPC)was used to assess the total changes in disease burden.The age-standardized mortality rate of intestinal infections in China decreased from 9.642/100 000 in 1990 to 0.439/100 000 in 2021,with an AAPC of-57.103%(95%CI:-57.118%to-57.088%).In 2021,Rotavirus,Norovirus,and Crypto-sporidium were the top three etiologies contributing to disease burden,with mortality rates of 1.020/100 000,0.040/100 000 and 0.079/100 000,respectively.A significant variation in etiology distribution was observed across age groups:Rotavirus,Shigella,and Crypto-sporidium dominated among children under 5 years of age,whereas Cryptosporidium,Norovirus,and Clostridioides difficile were more prevalent in older populations.Risk factor analysis indicated that unsafe water sources and poor sanitation accounted for 73.394%of all enteric disease-related deaths.In conclusion,the burden of enteric infections in China markedly declined from 1990 to 2021,and sig-nificant variations in the etiological spectrum and disease burden were observed across age groups.The persistent effects of unsafe wa-ter sources and poor sanitation underscore the need for targeted interventions to further decrease the burden of these diseases.Our find-ings highlight the success of public health interventions in decreasing the burden of enteric infections in China,while emphasizing the need for targeted measures to address disparities in high-risk populations and improve environmental sanitation.

Objective:To investigate the ameliorative effects of N-acetyl-D-mannosamine(ManNAc) on glucose and lipid metabolic disorders in obese mice.Methods:In vivo experiments were conducted using 21 four-week-old C57BL/6JGpt mice, randomly divided into three groups( n=7 per group): a normal control group, a high-fat diet(HFD) control grooup, and a ManNAc treatment group(400 mg·kg -1·d -1). The intervention lasted for 20 weeks. Body weight, food intake, and fasting blood glucose levels were monitored weekly. Glucose tolerance tests(GTT), insulin sensitivity tests(ITT), and respiratory metabolism monitoring were performed in the 17th, 18th, and 19th weeks, respectively. At the end of the experiment, whole-body fat distribution was assessed, and serum lipid profiles were measured. Liver and adipose tissue weights were recorded, and histological analyses including HE staining of liver, adipose and pancreatic tissues were performed. Liver transcriptome sequencing and quantitative real-time PCR(qPCR) were conducted to evaluate hepatic gene expression. In vitro, a hepatic steatosis model was established by inducing HepG2 cell with 0.4 mmol/L oleic acid, followed by treatment with 500 μg/mL ManNAc. Lipid accumulation was assessed using BODIPY staining, and the expression of lipid metabolism-related genes was quantified by qPCR. Results:ManNAc administration attenuated HFD-induced weight gain, reduced total body fat volume, and decreased liver and adipose tissue weights as well as intracellular lipid accumulation. Pancreatic islet numbers increased, while fasting blood glucose levels, glucose tolerance, and insulin sensitivity significantly improved. Serum levels of triglycerides, total cholesterol, and low-density lipoprotein levels were decreased, accompanied by enhanced energy expenditure. Additionally, hepatic expression of Cd36, Fabp3, and Scd1 was downregulated. In vitro, ManNAc significantly reduced lipid accumulation in HepG2 cells and downregulated the expression of Cd36, Fabp3, and Scd1 genes.Conclusion:ManNAc may improve glucose and lipid metabolism by modulating the PPARs-mediated fatty acid metabolic pathway, reducing lipogenesis, promoting fatty acid oxidation and energy expenditure, and enhancing insulin sensitivity, ultimately ameliorating disorders in obese mice.

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.