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.

An assay was established for detection of toxigenic Clostridioides difficile by combining microfluidic chip analysis with immunochromatography,and its performance was evaluated and compared with those of the Xpert C.difficile/Epi and VIDAS CD AB tests.Primer pairs were designed according to the tcdB and tpi genes in C.difficile.The specificity,limit of detection,reproducibility,and stability were evaluated.A total of 215 stool samples from patients with diarrhea were collected and tested in parallel with the Xpert C.difficile/Epi,VIDAS CDAB,and our assay.C.difficile was isolated from samples,and the tcdB gene was identified when discrepant results were obtained from the three above assays.Our assay showed no cross-reaction with other diarrhea-associated pathogens.Its reproducibility was 100％in testing of two standard plasmids containing tcdB and tpi genes at two concentrations(105 and 102 copies/μL).Two standard plasmids were detected after the PCR and immunochromatography reagents had been stored for 3,6,9,and 12 months,and all the results were posi-tive.The limit of detection was 10 copies/μL for toxigenic C.difficile.Testing of 33 samples positive for C.difficile with our assay(33/215,15.3％)yielded findings statistically coherent with those of the Xpert C.difficile/Epi test(kappa value=0.965).The sensitivity,specificity,positive predictive value,and negative predictive value of our assay,with respect to Xpert C.difficile/Epi as the standard,were 94.3％,100.0％,100.0％,and 98.9％;these values were significantly higher than those of VIDAS CDAB(60.0％,98.9％,91.3％,and 92.7％)(Kappa=0.714,OR=157.50,95％CI:62.03-847.28,P=0.013).In conclusion,our newly developed assay is specific,stable,and reproducible,and may be used for rapid and accu-rate detection of toxigenic C.difficile.The assay could be used for C.difficile infection screening in outpatient and emergen-cy,community medical service center,and epidemiological settings.

Objective:To assess the relationship between basophil levels and mortality in patients with intra-abdominal infection.Methods:Information on patients with intraperitoneal infection admitted to the intensive care unit were extracted from the MIMIC database.A time-dependent Cox regression model was used to adjust for confounders associated with 28-day mortality.Propensity score matching(PSM)was used to balance the baseline differences be-tween groups with different basophil levels,and a restricted cube chart(RCS)was used to show the relationship between basophil count and 28-day mortality in patients with intra-abdominal infection.Results:A total of 4403 patients with intra-abdominal infection were enrolled in the MIMIC database.Patients with high basophil levels have lower mortality than those with low basophil levels.There was an L-shaped curve between basophil level and 28-day mortality,with a cut-off value of 0.47×109/L.Cox regression analysis showed that basophil levels were an independent protective factor for mortal-ity in patients with intra-abdominal infection after adjusting for potential confounders(HR=0.586,95%CI:0.443-0.769).Protective factors for death at basophil levels remained after PSM adjusted for potential confounders(HR=0.628,95%CI:0.470-0.832).Conclusion:Basophil level is an independent protective factor for mortality in patients with intra-abdominal infection,and basophil levels should be dynamically monitored to better evaluate the prognosis of patients.

Objective To explore the effects of dodecanoylcarnitine(DA)and myristoleic acid(MA)on the function of mouse alveolar epithelial cell line MLE-12 and their underlying mechanisms.Methods An inflammatory model was established by stimulating MLE-12 cells with IL-4.The expression levels of DA,MA,and sphingosine-1-phosphate(S1P)in the cell supernatant were detected by ELISA.MLE-12 cells were separately intervened with DA and MA.RT-PCR and flow cytometry were used to detect the expression changes of inflammatory factors IL-6 and tumor necrosis factor-α(TNF-α)and the level of intracellular reactive oxygen species(ROS).Additionally,Western blot was performed to detect the expression of key proteins such as p38 mitogen-activated protein kinase(p-38 MAPK)and src homology 2 domain-containing phosphatase 1(SHP-1).To explore the role of S1PR2 in the effects of DA and MA,MLE-12 cells were pretreated with the S1PR2 inhibitor JTE-013,and the above experiments were repeated.Results IL-4 stimulation significantly upregulated the levels of DA,MA,and S1P in MLE-12 cells(P<0.05).DA/MA treatment groups exhibited significantly increased expression of IL-6 and TNF-α compared with the control group(P<0.05),along with elevated ROS levels(P<0.05).Western blot analysis revealed that DA/MA promoted SHP-1 dephosphorylation and phosphorylated p38 MAPK activation in MLE-12 cells.Notably,JTE-013 pre-treatment completely reversed these effects(P<0.05).Conclusion Asthma-related metabolites DA and MA exacerbate the inflammatory and oxidative stress responses of MLE-12 cells by activating the S1PR2 receptor,promoting the dephosphorylation of SHP-1 and the activation of the p-p38 MAPK pathway.This study reveals the core regulatory role of S1PR2 in this pathway as well.

Objective:To analyze the effects of the Focused Solution Model Nursing intervention on quality of life,negative emotions of the patients with prostate cancer.Methods:A total of 82 prostate cancer patients who were diagnosed and treated at the General Hospital of Eastern Theater Command between September 2022 and September 2024 were included and randomly divided into study group and control group by the method of random number table,with 41 patients in each group.The patients in the study group were treated with Focused Solution Model Nursing intervention.And the routine care was used in the control group The quality of life and negative emotions were compared between the two groups by using the scales of World Health Organization Quality of Life-Brief(WHOQOL-BREF),HAMA and HAMD.Results:Compared to the control group,the patients in the study group exhibited signifi-cantly higher scores in the physiological,psychological,environmental,and social relationship domains of the WHOQOL-BREF scale(P＜0.05).The scores of HAMA and HAMD in study group were lower than those of the control group(P＜0.05).Additionally,all subscales of the Social Impact Scale including social exclusion,internalized shame,social isolation and economic discrimination were significantly lower than those of the study group(P＜0.05).Conclusion:Focused Solution Model Nursing intervention can effec-tively improve the quality of life and negative emotions of the prostate cancer patients in the clinical treatment.

Objective To analyze the risk factors of delayed wound recovery after Meek skin grafting in patients with extensive deep burn and its predictive efficiency.Methods A total of 100 patients with extensive deep burn who underwent Meek skin grafting from August 2018 to November 2024 were selected,and they were divided into the normal group(n=79)and the delayed group(n=21)according to the wound healing time.The clinical data of all patients were collected,and the influencing factors of delayed wound recovery after Meek skin grafting in patients with extensive deep burn were analyzed by Logistic regression model,and the efficiency of the model was analyzed by the receiver operating characteristic(ROC)curve and the Hosmer-Lemeshow goodness-of-fit test,respectively.Results There were statistically significant differences in the body mass index,hospitalization time,burn index,burn area,donor skin area,number of postoperative dressing change,postoperative nutritional support,postoperative pain scores,and hospitalized blood glucose levels between the two groups(P<0.05).Logistic regression analysis showed that high burn index(OR=1.086,β=0.082),large burn area(OR=1.155,β=0.144),fewer postoperative dressing change(OR=0.746,β=-0.293),lack of postoperative nutritional support(OR=6.439,β=1.862),high postoperative pain score(OR=4.483,β=1.500),and high level of hospitalized blood glucose(OR=2.251,β=0.811)were the influencing factors for delayed wound recovery after Meek skin grafting in patients with extensive deep burn(P<0.05).The ROC curve revealed that the combined prediction of the above six influencing factors for delayed postoperative wound recovery had an area under the ROC curve(AUC)of 0.896,with a sensitivity and specificity of 88.00%and 89.10%,respectively;and the Hosmer-Lemeshow test result showed:χ2=10.641,and P=0.223,indicating a high predictive efficacy and a certain calibration ability of the model.Conclusion The high burn index,large burn area,fewer postoperative dressing change,lack of postoperative nutritional support,high postoperative pain score and hospitalized blood glucose level were the influencing factors for delayed wound recovery after Meek skin grafting in patients with extensive deep burn,and the prediction model constructed by the above factors has good predictive ability,which can provide a reference for clinical treatment.

Objective To analyze the risk factors of delayed wound recovery after Meek skin grafting in patients with extensive deep burn and its predictive efficiency.Methods A total of 100 patients with extensive deep burn who underwent Meek skin grafting from August 2018 to November 2024 were selected,and they were divided into the normal group(n=79)and the delayed group(n=21)according to the wound healing time.The clinical data of all patients were collected,and the influencing factors of delayed wound recovery after Meek skin grafting in patients with extensive deep burn were analyzed by Logistic regression model,and the efficiency of the model was analyzed by the receiver operating characteristic(ROC)curve and the Hosmer-Lemeshow goodness-of-fit test,respectively.Results There were statistically significant differences in the body mass index,hospitalization time,burn index,burn area,donor skin area,number of postoperative dressing change,postoperative nutritional support,postoperative pain scores,and hospitalized blood glucose levels between the two groups(P<0.05).Logistic regression analysis showed that high burn index(OR=1.086,β=0.082),large burn area(OR=1.155,β=0.144),fewer postoperative dressing change(OR=0.746,β=-0.293),lack of postoperative nutritional support(OR=6.439,β=1.862),high postoperative pain score(OR=4.483,β=1.500),and high level of hospitalized blood glucose(OR=2.251,β=0.811)were the influencing factors for delayed wound recovery after Meek skin grafting in patients with extensive deep burn(P<0.05).The ROC curve revealed that the combined prediction of the above six influencing factors for delayed postoperative wound recovery had an area under the ROC curve(AUC)of 0.896,with a sensitivity and specificity of 88.00%and 89.10%,respectively;and the Hosmer-Lemeshow test result showed:χ2=10.641,and P=0.223,indicating a high predictive efficacy and a certain calibration ability of the model.Conclusion The high burn index,large burn area,fewer postoperative dressing change,lack of postoperative nutritional support,high postoperative pain score and hospitalized blood glucose level were the influencing factors for delayed wound recovery after Meek skin grafting in patients with extensive deep burn,and the prediction model constructed by the above factors has good predictive ability,which can provide a reference for clinical treatment.

OBJECTIVE To investigate the therapeutic mechanisms of electroacupuncture at"Tianshu"(ST25)and"Sanyinjiao"(SP6)acupoints combined with metformin in the treatment of type 2 diabetes mellitus(T2DM)using serum non-targeted metabolomics.METHODS Male SD rats were randomly divided into blank group,model group,metformin group,electroacupunc-ture group,and acupuncture-medicine combination(electroacupuncture combined with metformin)group.A type 2 diabetes model was established by high-fat diet combined with intraperitoneal injection of streptozotocin.The metformin group was treated with 250 mg·kg-1 metformin by gavage,the electroacupuncture group was treated with bilateral Tianshu and Sanyinjiao,the acupuncture-medicine combination group was treated with metformin by gavage combined with electroacupuncture,and the blank group and model group were treated with normal saline by gavage.All rats were treated 6 times a week for 7 weeks.After the intervention,the blood glucose level in the tail vein of the rats was measured using a blood glucose meter in the fasting state.The blood glucose levels of the rats were measured at 30,60,120,and 240 min after intraperitoneal injection of 50%glucose solution(4 mL·kg-1)to evaluate glu-cose tolerance.The serum insulin level of the rats was detected by ELISA and the insulin resistance index was calculated.The blood biochemical parameters were measured by an automatic blood biochemical analyzer.HE staining was used to evaluate the pathological conditions of the liver and pancreatic tissues of the rats.Ultra-performance liquid chromatography-mass spectrometry(UPLC-MS)technology was used for mass spectrometry detection to identify differential metabolites,and MetaboAnalyst 5.0 was used for pathway enrichment analysis.RESULTS Compared with the blank group,the fasting blood glucose,area under the glucose tolerance curve,and insulin resistance index of the model group rats were significantly increased(P<0.001),blood TP and GLB were significantly de-creased(P<0.01),AST,ALT,and ALP were significantly increased(P<0.05,P<0.01,P<0.001),and obvious inflammatory cell infiltration and pathological damage were observed in the liver and pancreas tissues;compared with the model group,the fasting blood glucose,area under the glucose tolerance curve,and insulin resistance index of the acupuncture-medicine combination group were sig-nificantly decreased(P<0.05,P<0.01,P<0.001),blood ALP was significantly decreased(P<0.01),TP and GLB were significant-ly increased(P<0.05),and the pathological damage of the liver and pancreas was significantly improved.Serum metabolomics showed that the metabolic profiles of the groups were well distinguished.Compared with the blank group,the differential metabolites in the model group were enriched in histidine metabolism,thiamine metabolism,taurine and hypotaurine metabolism,ascorbic acid and alde-hyde ester metabolism,valine,leucine and isoleucine biosynthesis pathways;compared with the model group,237 metabolites such as 3-aminoadipic acid,3-oxocyclobutanecarboxylic acid and phosphorylcholine in the acupuncture-medicine combination group were sig-nificantly reduced,and the pathways were enriched in histidine metabolism,linoleic acid metabolism,thiamine metabolism,taurine and hypotaurine metabolism,valine,leucine and isoleucine biosynthesis pathways.CONCLUSION Electroacupuncture combined with metformin can effectively improve the glucose and lipid metabolism of T2DM rats,and its potential mechanism may be related to the regulation of amino acid metabolism.

Collagen is a matrix protein essential for maintaining the function of various tissues in ani-mals.There are many types of collagen in vertebrates,and the function of each type of collagen in the body is closely related to its sequence and structure.In addition to the common Gly-X-Y amino acid se-quence,there is also a special structure Gly-Gly-Y in the natural collagen.In order to explore its effect on collagen,this study constructed mutants containing Gly-Gly-Y at the levels of collagen polypeptides,long-chain collagen,and collagen polymers.The thermal stability of the mutant before and after mutation was characterized by circular dichroism scanning.At the same time,molecular dynamics simulation was used to calculate the hydrogen bonding probability and bending degree of collagen polypeptides,explai-ning the molecular mechanism of changes in collagen stability and flexibility.The results showed that the mutant containing the Gly-Gly structure would decrease the Tm value of the sample,but this effect would gradually weaken as the collagen triple helix region lengthened and the degree of protein self-assembly in-creased,and the reduced Tm values are 5 ℃,3 ℃,and 1 ℃,respectively.At the same time,the sim-ulation results show that the curvature of the polypeptide containing the Gly-Gly structure also increases to some extent,indicating that the structure near the mutation site has greater flexibility.This provides a new idea for the design of rubber materials with certain flexibility.

Objective To analyze the medication rules and clinical experience of Qin Zujie in treating virtual labor based on data mining.Methods Outpatient medical records of Qin Zujie,diagnosed with deficiency syndrome at Ethnic Medicine Characteristic Diagnosis and Treatment Center,Guangxi International Zhuang Medical Hospital from January to December 2024 were selected.A standardized database was established and processed,yielding 148 clinical prescriptions containing 171 herbal ingredients.Frequency analysis,association rule analysis,and cluster analysis were employed to examine medication rules and clinical experience.Results The 20 Chinese herbal ingredients used more than 30 times each,with the first three being Zhigancao,Baizhu,Danggui.These herbs are primarily warm in nature,neutral in temperature,and slightly cold in effect,characterized by sweet,pungent,and bitter flavors.They mainly affect the spleen meridian and function to tonify deficiency,with Qi-tonifying herbs being the most common.Through association rule analysis,the most frequently combined pairs were Baizhu-Dangshen.Cluster analysis identified three core prescription clusters.Conclusion Qin Zujie's academic thought of treating virtual labor is to supplement deficiency as the main,detoxification as the auxiliary,and treat both the symptoms and the causes.