OBJECTIVE To explore the changes and correlations of color， components and anti-tumor activity of Gelsemium elegans during sand-frying processing， so as to provide a reference for the toxicity-attenuating and efficacy-preserving processing as well as quality control of sand-fried G. elegans decoction pieces.METHODS Raw G.elegans and its processed products prepared by sand-frying for 15， 30， 75， 105 and 120 s were taken as test samples. The chromatic values， contents of total alkaloids and seven main active components （gelsemine， humantenine， koumine， gelsenicine， etc.） were determined. Their in vitro anti-tumor activity was investigated. The correlations between color and component contents， as well as between total alkaloid content and anti-tumor activity were analyzed.RESULTS The color of G.elegans samples generally darkened after sand-frying， and the total color difference （Δ E * ） of samples sand-fried for 75 s was greater than 20. At the initial stage of sand-frying （15-30 s）， the total alkaloid content in processed products decreased sharply compared with the raw material； at the later-stage of sand-frying （75-105 s）， the declining rate of total alkaloid content slowed down and tended to be stable. The contents of gelsenicine and humantenidine in processed products dropped drastically at 30 s of sand-frying， while the contents of koumine and sempervirine decreased substantially （by about 80%） after 75 s of sand-frying and then gradually tended to be stable. The anti-HepG2 cell proliferation activity of each processed product decreased with the extension of frying time. Correlation analysis demonstrated that color brightness and yellow-blue chromaticity were significantly positively correlated with the contents of total alkaloids and all determined active components except gelsemine （ P ＜0.05 or P ＜0.01）； red-green chromaticity was significantly positively correlated with the contents of koumine， sempervirine， gelsemivirine and humantenidine （ P ＜0.05 or P ＜0.01）； Δ E * was significantly negatively correlated with the contents of total alkaloids and all determined active components except gelsemine. Besides， total alkaloid content was significantly negatively correlated with the half maximal inhibitory concentration of the samples on HepG2 cells （ P ＜0.05 or P ＜0.01）.CONCLUSIONS With the prolongation of sand-frying time， the color of sand-fried G.elegans decoction pieces gradually turned brownish-brown， the contents of seven active components and total alkaloids decreased progressively， and its anti-HepG2 cell activity showed an overall downward trend. Sand-frying for 75 s serves as a critical turning-point for the above-mentioned changes.

Objective To quantitatively evaluate structural changes in the TMD disc using magnetic resonance T2 mapping technology at open and closed mouth positions,and assess its significance.Methods 127 newly diagnosed patients with TMD and 51 controls in the Department of Oral and Maxillofacial Surgery of the Affiliated Hospital of Yunnan University from June 2023 to January 2024 were prospectively collected.All subjects underwent routine TMJ sequence and T2 mapping sequence scans.After scanning,the pseudo-color images were processed,and T2 values of the joint disc(anterior and posterior bands)were measured at open and closed mouth positions to analyze their correlation with TMD disc displacement.Results The T2 values of the posterior zone of the joint disc in both open and closed positions were higher than those in the anterior zone of the joint disc in the case group(P<0.05);for different anterior displacement degrees of the joint disc,T2 values showed a progressive increase from the normal position group to the mild and moderate anterior displacement groups,while the severely displaced group demonstrated a decreasing trend(P<0.01);joint clicking and occlusal abnormalities were independent risk factors for non-reducible anterior joint disc displacement in TMD patients.Conclusion Scans during both mouth-opening and closing positions can be used to assess joint disc injury in TMD;T2 mapping technology can quantitatively and sensitively reflect changes in the biochemical composition of joint disc microstructure.

Objective To analyze the risk factors affecting the progression-free survival(PFS)of patients with intrahepatic cholangiocarcinoma(ICC)after surgical resection,and a predictive nomogram model was constructed based on blood test indicators.Methods A total of 99 ICC patients who received their first treatment at Hunan Provincial People's Hospital from January 2020 to December 2022 were selected as research subjects,they were divided into training group(n=70)and validation group(n=29).The independent risk factors for postoperative recurrence were identified using least absolute shrinkage and selection operator(LASSO)regression analysis and multivariate Cox risk regression analysis.The accuracy and clinical utility of the method were further tested by consistency index,receiver operating characteristic(ROC)curve,calibration graph,decision curve analysis,and progression-free survival curve.Results LASSO and multivariate Cox risk regression analysis showed that patient's gender,tumor-lymph node metastasis,differentiation grade,tumor diameter,aspartate aminotransferase,alpha-fetoprotein,total bile acid after surgery,and monocyte changes before and after surgery were independent factors affecting the PFS of ICC patients.The predictive model based on the risk factor showed that the consistency index of the training group was 0.884 and that of the validation group was 0.838.The area under ROC curve for training group:PFS prediction at 6 months,1 year,and 2 years were 0.972,0.965 and 0.923,respectively;for validation group:PFS prediction at 6 months,1 year,and 2 years were 0.972,0.821 and 0.923,respectively.The risk score calculated according to the nomogram divided ICC patients into high-risk group and low-risk group with high postoperative recurrence risk,and the median PFS in high-risk group was significantly shorter than that in low-risk group(P＜0.05).Conclusion The nomogram constructed by this study has good predictive effect and can be used as a supplementary evaluation for predicting PFS after ICC patients.

Objective Analysing the risk factors for the occurrence of pre-frailty or frailty of the community elderly in Chengdu and constructing a risk prediction model.Methods The general information questionnaire,MNA-SF scale,Changsha version of the MoCA scale,PSQI scale,FRAIL scale,and Traditional Chinese Medicine Constitution Scale for the elderly were used in the cross-sectional survey.A total of 400 elderly people who completed community physical examinations in Chengdu from April 2022 to April 2023 were selected as the research objects.Based on multivariate Logistic regression analysis,the independent influencing factors were determined,and RStudio software was used to construct a risk prediction model nomogram.The physical examination data of 200 elderly people collected in Deyang City from June 2023 to October 2023 were used for external verification.The area under the ROC curve,Hosmer-Lemeshow test and calibration curve,and decision curve were used to evaluate the discrimination,calibration,and clinical practicability of the model.Results Age,abdominal circumference,number of chronic diseases,PSQI score,allergy history,and balanced constitution were independent influencing factors for pre-frailty/frailty in the elderly in the community(P<0.05),and the regression equation was as follows:Logit(P)=0.063×age+0.025×abdominal circumference-1.006×allergy history+0.300×number of chronic diseases+0.082×PSQI-1.013×balanced constitution-8.269.The area under the ROC curve of the modeling group was 0.779(95%CI:0.733-0.825),the sensitivity was 67.1%,the specificity was 76.3%,and the maximum Youden index was 0.434.The area under the ROC curve of the external validation group was 0.783(95%CI:0.709-0.856),the sensitivity was 62.0%,the specificity was 95.0%,and the maximum Youden index was 0.570.The Hosmer-Lemeshow goodness of fit test results of the two groups were χ2=3.285,P=0.915 and χ2=8.376,P=0.398,and the calibration curve fit was good.DCA showed that the threshold probability of clinical benefit was 5%-99%and 21%-98%.Conclusions The pre-frailty/frailty risk prediction model established in this study has good predictive efficacy for the elderly in the community,and the use of this model for screening and early intervention of high-risk populations can be clinically beneficial.

Objective:To establish a gas chromatography-mass spectrometry method(GC-MS-MS)for determining the content of methyl p-toluenesulfonate(MTS),ethyl p-toluenesulfonate(ETS),and isopropyl p-toluenesulfonate(IPTS)in tosufloxacin tosylate hydrate.Methods:The HP-1MS(0.250 mm ×30 m,0.50 μm)column was used at progamming temperature,the injection temperature was 250 ℃.The MS conditions were as follow:the ionization mode was EI,the electron voltage was 70 V,the ion source temperature was 250 ℃,the scanning method was MRM,the quantitative ion pairs for MTS,ETS and IPTS were m/z 91 →65,m/z 155→91 and m/z 91→65.Results:The linearity ranges of MTS,ETS and IPTS were 48.779-975.59 ng·mL-1(r=0.998 5),54.586-1 091.7 ng·mL-1(r=0.998 4)and 46.241-924.82 ng·mL-1(r=0.999 8).The average recoveries were 99.47％,99.15％,98.83％(n=9).The MTS,ETS and IPTS were not detected in the 7 batches of tosufloxacin tosylate hydrate.Conclusion:The established method can be used for the determination of MTS,ETS,and IPTS content in tosufloxacin tosylate hydrate.

This study was aimed atanalyzing the epidemic characteristics of dengue fever in Ruili City atthe China-Myanmar bor-der in late 2023,to provide evidence for local dengue fever prevention and control measures.Adult Aedes mosquitoes were collected from Ruili City with a backpack type mosquito sucking machine in October of 2023.Serum samples frompatients with suspected den-gue were collected in acutephase,in November of 2023.Detection ofdengue virus(DENV)nucleic acids in Aedesmosquitoes and acute phase serum samples from suspected dengue fever patients using reverse transcription-polymerase chain reaction,and nucleic acid positive samples were inoculated into Vero cells for viral culture.After three consecutive blind passage,samples with cytopathic effect(CPE)were collected for be sequencingand analysisof genetic and evolutionary information.Dengue case characteristics were analyzed through descriptive epidemiological methods.Among the 109 cases of dengue fever,the ratio of males to females was 1.27∶1.The youngest patient was 1 year old,the oldest patient was 84 years old,the age group of 20～59 years accounted for 73.39%,and the major-ity of occupations were freelancer(40.37%).A total of 827 female Aedes albopictus and 312 Aedes aegypti were collected,all of which tested negative for DENV nucleic acid.109 serum samples tested positive for DENV nucleic acid,including 49 DENV-1 and 60 DENV-2.Moreover,five DENV-1 and nine DENV-2 samples were obtainedthrough third-generation blind passaging with CPE.The E gene sequences of these five DENV-1 strains were detected,all were found to belong to DENV-1 genotype I,and had same evolu-tionary branch as the 2023 Guangzhou,China(PP563911),the 2019 Myanmar(MW793710),and 2019 Attapeu,Laos(MW559046).The nucleotide and amino acid sequence similarityamong the five DENV-1 genotype was 99.4%-99.9%and 99.8%-100.0%.Compared with the aforementioned epidemic strains,their nucleotide similarities were 99.5%-100.0%,99.4%-99.6%and 99.3%-99.5%,respec-tively,and amino acid similarity was 99.8%-100.0%.Nine DENV-2 E gene sequences were of Asian genotype I and belonged to the same evolutionary branch as the 2018 Myanmar(MW788982),2019 Hangzhou(OP684212)and 2019 Ruili(OQ928150).The nucleotide and amino acid similarities of the nine samples were 99.5%-100.0%and 99.8%-100.0%,respectively.Compared with the aforementioned epidemic strains,their nucleotide similarities were 99.7%-100.0%,99.3%-99.7%and 99.3%-99.7%,respectively,and the amino acid similarity was 99.8%-100%,99.8%-100.0%and 99.4%-99.6%,respectively.Two dengue vectors,Aedes albopictus and Aedes aegypti,were present in Ruili city,and the dengue outbreak was caused primarily by DENV-1 genotype I and DENV-2 Asian genotype I in later 2023.The sources of DENV-1 were probably the same as those of DENV-1 with Guangzhou(2023),and the sources of DENV-2 were probably from Myanmar.Dengue cases were found primarilyin the 20-59 year age group and freelancers,thus suggesting that relevant local departments should strengthen surveillance of dengue imported case and vector.

ObjectiveTo explore the current situation of forensic ethics practice and education by designing a questionnaire on forensic ethics， with a view to exploring the path of forensic ethics education construction. MethodsA total of 667 valid questionnaires were collected using the online survey method， basically covering various regions across the country and all sub-specialties of forensic medicine. Descriptive analysis was used to analyze the relevant data. ResultsMost practitioners had relevant ethical reflections in the process of forensic practice. 69.12% of the respondents indicated that they had studied the relevant rules， but approximately half stated that there were no corresponding ethical norms or standard operating manuals. The specific behaviors violating ethics in different units were diverse. 23.04% of the respondents reported that they had encountered unethical behaviors， but only 4.9% of them reported such violations. In terms of forensic ethics education， 87.75% of the respondents believed that there were issues with the current model of forensic ethics education. Meanwhile， the respondents showed a high degree of recognition for receiving forensic ethics education， with 84.15% of respondents expressing willingness to participate in relevant courses. More than half of respondents were willing to participate in forensic ethics education during undergraduate studies， new employee training， and regular post-employment training. ConclusionCurrently， there is a problem of ethical neglect in forensic work in China. Combining ethics courses with professional courses at the practitioner training stage and providing regular training at the practice stage are effective measures to popularize forensic ethics knowledge， enhance ethical awareness， and improve the quality of practice.

OBJECTIVE: To elucidate the role and underlying mechanism of ubiquitin-specific protease 35 (USP35) in ferroptosis of rheumatoid arthritis-fibroblast like synoviocytes (RA-FLS), thereby enhancing our comprehension of the pathogenesis of RA and identifying potential therapeutic targets for its treatment. METHODS: (1) RA-FLS were cultured in vitro and transduced with lentiviral vectors to establish stable cell lines: A USP35-knockdown line (short hairpin ribonucleic acid of USP35, shUSP35) and its control (negtive control of short hairpin ribonucleic acid, shNC), as well as a overexpression of USP35 line (USP35 OE) and its control (Vector). To investigate the role of USP35 in ferroptosis regulation, a ferroptosis model was induced in RA-FLS by treatment with 1 μmol/L Erastin. The cells were divided into six groups: shNC, shNC + Erastin, shUSP35 + Erastin, Vector, Vector + Erastin, and USP35 OE + Erastin. (2) Cell viability was detected using the cell counting kit-8 (CCK-8). (3) Reactive oxygen species (ROS), malondialdehyde (MDA), glutathione/glutathione disulfide (GSH/GSSG) ratios, and Ferrous ion (Fe²⁺) levels were measured using specific assay kits to evaluate oxidative stress, lipid peroxidation, and glutathione redox status in the cells. (4) Protein expression levels of solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4) were detected using Western blotting to investigate their potential involvement in USP35-mediated ferroptosis regulation. RESULTS: (1) Compared with the shNC +Erastin group, the cell viability of the shUSP35+Erastin group was significantly decreased (P < 0.001), while it was notably increased in the USP35 OE+Erastin group compared with the Vector+Erastin group (P < 0.001). These findings indicated that USP35 could alleviate the inhibitory effect of Erastin on RA-FLS cell viability. (2) In comparison to the shNC+Erastin group, the levels of ROS (P < 0.001), MDA (P < 0.05), and Fe²⁺ (P < 0.001) were significantly elevated, and the GSH/GSSG ratio was increased (P < 0.05) in the shUSP35+Erastin group. Conversely, the levels of ROS (P < 0.001), MDA (P < 0.05), and Fe²⁺ (P < 0.05) were significantly decreased, and the GSH/GSSG ratio was decreased (P < 0.05) in the USP35 OE+Erastin group compared with the Vector+Erastin group. These results suggested that USP35 could inhibit Erastin-induced oxidative stress and lipid peroxidation in RA-FLS. (3) In Erastin-induced RA-FLS, the expression of USP35 was positively correlated with the protein levels of SLC7A11 and GPX4, indicating a potential mechanism by which USP35 regulated ferroptosis in these cells. CONCLUSION USP35 inhibits ferroptosis in RA-FLS, potentially through the increased expression of SLC7A11 and GPX4.
Ferroptosis ; Humans ; Arthritis, Rheumatoid/metabolism* ; Synoviocytes/pathology* ; Reactive Oxygen Species/metabolism* ; Ubiquitin-Specific Proteases/metabolism* ; Fibroblasts/pathology* ; Cell Survival ; Piperazines/pharmacology* ; Endopeptidases/metabolism* ; Cells, Cultured ; Cell Line ; Amino Acid Transport System y+

Objective To investigate the correlation of serum levels of soluble programmed death ligand 1(sPD-L1),tumor necrosis factor-associated apoptosis-inducing ligand(TRAIL)and fibroblast growth factor 23(FGF-23)with risk stratification and death in patients with acute pulmonary embolism(APE).Methods A total of 113 pa-tients with APE admitted to the hospital from January 2022 to January 2024 were selected as APE group,and 50 healthy subjects were selected as control group.The 113 patients with APE were divided into high risk group(39 cases),medium risk group(45 cases)and low risk group(29 cases)by risk stratification.Accord-ing to the death of APE patients,they were divided into survival group(83 cases)and death group(30 cases).The levels of serum sPD-L1,TRAIL and FGF-23 were measured by enzyme-linked immunosorbent assay(ELISA).Multiple Logistic regression was used to analyze the risk factors affecting death of APE patients.Receiver operating characteristic(ROC)curve was drawn to analyze the value of serum sPD-L1,TRAIL and FGF-23 levels in evaluating mortality in APE patients.Pearson correlation analysis was used to investigate the correlation between serum sPD-L1,TRAIL and FGF-23 levels and cardiac function indicators in APE patients.Results The levels of serum sPD-L1,TRAILand FGF-23 in the APE group were significantly higher than those in the control group(P＜0.001).The serum sPD-L1,TRAIL and FGF-23 levels in the death group were significantly higher than those in the survival group(P＜0.001).The levels of serum sPD-L1,TRAIL and FGF-23 in the high-risk group were higher than those in the medium and low-risk groups,and the medium risk group was higher than the low-risk group,and the differences were statistically significant(P＜0.001).Multiple Logistic regression analysis showed that elevated levels of cardiac troponin Ⅰ(cTnⅠ),B type brain na-triuretic peptide(BNP),sPD-L1,TRAIL and FGF-23 were risk factors for mortality in APE patients(P＜0.05).ROC curve analysis showed that the combination of sPD-L1,TRAIL and FGF-23 had the largest area under curve for predicting death in APE patients,which was 0.924(95％CI:0.861-0.986),with a sensitivity of 96.8％and a specificity of 81.2％.Correlation analysis showed that the levels of serum sPD-L1,TRAIL and FGF-23 were positively correlated with cTnⅠ and BNP in APE patients(P＜0.001).Conclusion The lev-els of serum sPD-L1,TRAIL and FGF-23 are significantly elevated in APE patients,and their high expression is associated with high-risk stratification and mortality.The combination of these three indexes has good eval-uation value for APE patient mortality.