ObjectiveTo analyze the pharmacodynamic substance basis of Shangkeling Spray and its potential mechanisms in intervening knee osteoarthritis （KOA） using ultra-performance liquid chromatography-tandem mass spectrometry （UPLC-MS）， network pharmacology， and molecular docking technology. MethodsUPLC-MS was used to identify the chemical components of Shangkeling Spray. Pharmacokinetic properties were employed to screen potential active ingredients. Network pharmacology methods were utilized to collect potential targets of these ingredients and the pathological gene set of KOA. An "active ingredient-disease" target network was constructed using databases such as STRING. Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） functional enrichment analyses were performed using clusterProfiler. Libraries including NumPy were employed to calculate shortest path lengths to identify dominant pharmacodynamic links. Core gene clusters were identified using MCODE， validated through the Gene Expression Omnibus （GEO） database， and molecular docking was performed between key active ingredients and core targets. ResultsA total of 322 and 314 chemical components were identified under positive and negative ion modes， respectively， with 410 components in total after de-duplication， mainly including flavonoids， coumarins， terpenoids， organic acids， and alkaloids. Analysis of the "active ingredient-disease" network identified "development and regeneration"， "cell growth and death"， "immune system"， and "nervous system" as the dominant pharmacodynamic links of Shangkeling Spray in the treatment of KOA. Molecular docking showed that key active ingredients， such as bletillin A， formononetin， morin， oxymatrine， aconitine， gallic acid， curdione， apigenin， naringenin， and oleanolic acid， tightly bound to functional domains of 10 key targets including Jun proteins（JUN）， interleukin-6 （IL-6）， protein kinase B1 （Akt1）， Caspase-3， nuclear transcription factor-κB subunit p65（RELA）， nuclear factor-kappaB1（NF-κB1）， Cyclin D₁， mammalian target of rapamycin（mTOR）， tumor necrosis factor （TNF）， and Fos proto-oncogene protein （FOS）. These interactions synergistically regulated the phosphatidylinositol 3-kinase （PI3K）/Akt/mTOR-related signaling axis and nervous system-related pathways， mediating cartilage repair， reducing inflammation and pain， and improving KOA. ConclusionThis study preliminarily clarifies the pharmacodynamic substance basis of Shangkeling Spray and suggests that its main active ingredients may improve KOA by synergistically regulating the PI3K/Akt/mTOR-related pathways， providing a reference for subsequent exploration of its substance benchmark and mechanism of action.

ObjectiveTo explore the effect and mechanism of modified Dingjingtang in regulating the Toll-like receptor 2 （TLR2）/nuclear factor （NF）-κB/NOD-like receptor protein 3 （NLRP3） signaling pathway to inhibit inflammatory skin lesions in the rat model of acne. MethodsForty-eight rats were randomized into the normal， model， low-， medium-， and high-dose （8.1， 16.2， and 32.4 g·kg^-1） modified Dingjingtang， and doxycycline hydrochloride （0.27 g·kg^-1） groups， with 8 rats in each group. Rats in other groups except the normal group were modeled by intradermal injection and intraperitoneal injection of Propionibacterium acnes. After successful modeling， rats in the intervention groups were treated with corresponding agents by gavage， and those in the normal and model groups with an equal volume of normal saline， once a day for 14 consecutive days. Then， the samples were collected. The general conditions， ear thickness， and body weight changes of rats were observed. Biochemical methods were used to determine superoxide dismutase （SOD） and malondialdehyde （MDA） levels in the ear tissue. Hematoxylin-eosin staining and Masson's staining were used to observe the pathological changes and collagen deposition， respectively， in the ear tissue. Immunohistochemistry was employed to determine the interleukin （IL）-1β level in the ear tissue. Enzyme-linked immunosorbent assay was adopted to measure the levels of IL-1β， tumor necrosis factor （TNF）-α， and IL-6 in the serum. The total antioxidant capacity method was adopted to assess the reactive oxygen species （ROS） content in the ear tissue. Western blot and real-time fluorescence quantitative polymerase chain reaction were employed to determine the protein and mRNA levels， respectively， of TLR2， myeloid differentiation factor 88 （MyD88）， NF-κB， NLRP3， and cysteinyl aspartate-specific proteinase-1 （Caspase-1） in the ear tissue. ResultsCompared with the normal group， the model group had increased ear skin thickness （P<0.01）， elevated ROS and MDA levels （P<0.01）， reduced SOD content （P<0.05）， and increased collagen deposition （P<0.01） in the ear tissue. In addition， the model group showed raised IL-1β， IL-6， and TNF-α levels in the serum （P<0.01） and up-regulated mRNA and protein levels of TLR2， MyD88， NF-κB， NLRP3， and Caspase-1 （P<0.01）. Compared with the model group， the high- and medium-dose modified Dingjingtang groups showed significant improving effects regarding the above indicators （P<0.05， P<0.01）. ConclusionModified Dingjingtang can ameliorate the inflammatory skin lesions in the rat model of acne by regulating the TLR2/NF-κB/NLRP3 signaling pathway.

Objective To investigate the predictive value of the combined tumor burden score (TBS) and platelet-albumin-bilirubin (PALBI) score model for postoperative tumor recurrence in liver transplant recipients with hepatocellular carcinoma (HCC). Methods The general information of 158 recipients diagnosed with HCC and underwent liver transplantation at the 900th Hospital of the Joint Logistics Support Force of the Chinese People's Liberation Army from 2008 to 2021 was collected. Lasso regression analysis combined with multivariate Cox regression analysis were used to identify independent risk factors for postoperative tumor recurrence after liver transplantation with HCC. A nomogram prediction model was constructed based on variables selected by Lasso regression analysis, and the predictive performance of the model was verified by calibration curve and clinical decision curve. The optimal cut-off values for postoperative tumor recurrence in liver transplant recipients with HCC were determined by receiver operating characteristic (ROC) curve, and Kaplan-Meier analysis was used to compare survival differences among different groups. Results Among the 158 liver transplant recipients with HCC, 82 experienced tumor recurrence, with a recurrence rate of 51.9% and a median tumor-free survival time of 10 (4, 25) months. Results of Lasso regression analysis and multivariate Cox regression analysis showed that alpha-fetoprotein (AFP) ≥400 ng/mL, TBS and PALBI score were all independent risk factors for postoperative tumor recurrence in liver transplant recipients with HCC (all P<0.05). The combined high TBS-high PALBI score showed the highest predictive value (hazard ratio 6.909, 95% confidence interval 3.067-15.563, P<0.001). A nomogram prediction model was constructed based on six variables selected by Lasso regression analysis. Calibration curve showed good consistency between the model's predicted results and the ideal curve. Decision curve analysis indicated that the nomogram prediction model provided the highest clinical benefit for predicting 1-year tumor-free survival after liver transplantation with HCC. Time-dependent ROC curves at 1, 3 and 5 years after surgery showed that TBS-PALBI model had good predictive performance, with no significant difference in area under the curve (AUC) compared with TBS-PALBI-AFP model. The optimal cut-off values for predicting postoperative tumor recurrence were determined by ROC curve, with a PALBI score cut-off of −2.334 and a TBS cut-off of 5.305. Recipients were divided into a low TBS-low PALBI score group (n=47) and a low/high TBS-low/high PALBI score group (at least one score was high) (n=111). Kaplan-Meier survival analysis showed that the low TBS-low PALBI score group had a higher tumor-free survival rate than the low/high TBS-low/high PALBI score group, with a significant difference (P<0.05). Conclusions TBS-PALBI model provides a novel, simple and effective tool for assessing the prognosis of liver transplant recipients with HCC. The nomogram model constructed based on this has significant advantages in predictive performance and may serve as a reference for guiding individualized treatment plans and improving clinical outcomes.

Objective To investigate the efficacy and safety of the Corheart 6 left ventricular assist system in patients with end-stage heart failure. Methods A retrospective study was conducted on patients with end-stage heart failure who were treated with Corheart 6 left ventricular assist system from March 2022 to June 2024 in 4 hospitals in Jiangsu Province. The efficacy of the device was evaluated by comparing changes in clinical indicators at preoperative, discharge, 3-month postoperative, and 6-month postoperative timepoints, including the New York Heart Association (NYHA) functional classification, left ventricular ejection fraction (LVEF), and left ventricular end-diastolic diameter (LVEDD). The safety of the device was assessed by analyzing the intraoperative position and orientation of the blood pump inlet cannula, as well as the incidence of adverse events. Results In this study, 39 patients were collected, including 34 males and 5 females with a mean age of (56.4±12.5) years, ranging from 20 to 75 years. There was no operative death. There was no death in postoperative 3 months with a survival rate of 100.0%. There were 3 deaths in 6 months postoperatively, with a survival rate of 92.3%. All patients had a preoperative NYHA cardiac function classification of class Ⅳ. The NYHA cardiac function class of the patients improved (P<0.05) at discharge, 3 and 6 months after surgery when compared to the preoperative period. LVEF was significantly higher at 3 months after surgery than that during the preoperative period (P<0.05). LVEDD was significantly smaller at discharge, 3 and 6 months after surgery than that during the preoperative period (P<0.05). The safety evaluation's findings demonstrated that all 39 patients' intraoperative blood pump inlet tubes were oriented correctly, the artificial blood vessel suture sites were appropriate, there were no instances of device malfunction or pump thrombosis, or instances of bleeding or hemolysis, and the rate of the remaining adverse events was low. Conclusion With a low rate of adverse events and an excellent safety profile, the Corheart 6 left ventricular assist system can efficiently enhance cardiac function in patients with end-stage heart failure. It also has considerable clinical uses.

AIM To study the chemical constituents from the petroleum ether fraction of the roots of Gypsophila licentiana Hand.-Mazz.METHODS Silica gel,Sephadex LH-20 and semi-preparative HPLC were used for isolation and purification,then the structures of obtained compounds were identified by physicochemical properties and spectral data.RESULTS Eighteen compounds were isolated and identified as dibutyl phthalate(1),glyceryl arachidate(2),bis(2-ethylhexyl)terephthalate(3),9,12-octadecadienoic acid(Z,Z)-methyl ester(4),(3'S,4'S)-3'-acetoxy-4'-angeloyloxy-3',4'-dihydroseselin(5),3-(4-hydroxy-3-methoxyphenyl)-propanoic acid(6),bis(2-ethylhexyl)phthalate(7),2,2'-oxybis(1,4)-di-tert-butylbenzene(8),gypsogenin(9),3-keto,16α-hydroxy,24-noroleanolic acid(10),3-oxo-olean-12-en-28-oic acid(11),10-eicosenoic acid(12),hexacosanic acid(13),enniatin B(14),(R,Z)-21-methyl-8-pentatriacontene(15),ethyl gallate(16),stellarine A(17),pentacosane(18).CONCLUSION All compounds are isolated from this plant for the first time.

Objective To analyze the epidemiological and clinical characteristics of scrub typhus in Liuyang People's Hospital Hunan Province for better diagnosis,treatment,prevention and control of scrub typhus.Methods A retrospective study was conducted on 159 confirmed cases of scrub typhus.The demographic data of patients,clinical manifestations of scrub typhus,laboratory tests,and chest CT findings,complications,treatment,and outcomes were analyzed.Results The 159 patients with scrub typhus included 142 males and 17 females.The average age of patients was(53.8±11.9)(18-82)years old.The peak incidence of scrub typhus was in the period from July to September(87.42％).The common clinical manifestations included fever(97.48％),headache(41.51％),and fatigue(23.90％).The most common signs were eschar(92.45％)on the lower limbs(25.16％),scrotum(15.72％),and buttocks(11.32％).Laboratory test results upon admission showed decreased platelet count(64.15％),elevated creatinine(38.36％),elevated AST(90.57％),ALT(80.50％),and LDH(90.57％).Pulmonary imaging study revealed pulmonary inflammation(22.64％),pleural effusion(13.21％),and pericardial effusion(6.29％).The common complications included liver impairment(69.81％)and decreased platelet count(50.94％),as well as other complications such as renal dysfunction,myocardial injury,sepsis,respiratory failure,and disseminated intravascular coagulation(DIC).Doxycycline-based therapies resulted in cure rate of 98.11％(156/159).Three patients died due to severe complications.Conclusions The peak incidence of scrub typhus was in the period from July to September in Hunan Province.Eschar is a key feature for clinical diagnosis.Scrub typhus can affect multiple organ systems,leading to various systemic complications.The cure rate is high with doxycycline treatment,but some patients may die from severe complications due to delayed medical treatment.

Objectives:To compare the efficacy and imaging parameters of lateral lumbar interbody fusion(LLIF)combined with anterolateral screw fixation(AF)or bilateral pedicle screw fixation(BPSF)in the treatment of lumbar degenerative disease(LDD).Methods:A retrospective study was conducted on 100 patients with single-segment LDD who underwent LLIF-AF or LLIF-BPSF treatment at the First Affiliated Hospital of Kunming Medical University between December 2019 and December 2021.The patients were divided into the AF group(50 cases)and the BPSF group(50 cases).There was no statistical difference in the general informa-tion such as gender,age,and body mass index(BMI)between the two groups(P＞0.05).The perioperative data(length of hospital stay,operative time,intraoperative blood loss);Pre-operative,postoperative 7d,postoperative 6 months' visual analog scale(VAS)scores for low back and leg pain,Oswestry disablity index(ODI),imaging parameters such as disc height(DH),foraminal height(FH),cross-sectional area(CSA),as well as surgical complications were collected and analyzed,and the risk factors for intervertebral subsidence and non-fusion of fusion device were explored.Results:AF group was better than BPSF group in operative time,intraoperative blood loss,and hospital stay[125.0±26.6min vs 221.6±52.2min,25.0mL(20.0-50.0mL)vs 100.0mL(90.0-150.0mL),12.5±4.8d vs 14.9±4.6d],and the differences were statistically significant(P＜0.05).For the low back and leg pain VAS scores and ODI,as well as DH,FH,and CSA,the postoperative 7d and 6 months'values were signiticantly improved than before operation in both groups(P＜0.05),while no statistically significant differences were found between groups at the same time points(P＞0.05).Intervertebral subsidence occurred in 12 patients in each group,and there was no statistically significant difference between the two groups.There were statistically significant differences in BMI and QCT between the AF and BPSF groups of patients with subsidence of the intervertebral space and those without subsidence at 6 months after operation;There were statistically significant differences in QCT between the two groups of patients with non-fusion and fusion of the fusion device;BMI≥28kg/m2 and QCT＜80mg/cm3 were the independent risk factors for intervertebral subsidence in AF group,but not in the BPSF group;QCT＜80mg/cm3 was the independent risk facors for non-fusion of fusion device in both AF and BPSF groups.Conclusions:LLIF combined with AF or BPSF are both reliable methods for treating LDD.For patients with a high body weight of BMI≥28kg/m2 or decreased bone density of QCT＜80mg/cm3,BPSF internal fixation can provide stronger mechanical stability to the spine,reducing the incidence of postoperative disc space collapse or fusion device non-union;On the contrary,AF internal fixation has shorter operative time and hospital stay,less intraoperative blood loss,which can be considered as a priority.

This study was aimed at exploring the interaction between the nucleoprotein(NP)of influenza A virus(IAV)and TRIM25.The physicochemical properties and protein structure of IAV NP protein were analyzed through bioinformatics methods.The interaction between IAV NP and TRIM25 proteins was simulated with molecular docking techniques,and the in-teraction sites were predicted.With the cDNA of the A/Puerto Rico/8/1934(H1N1)PR8 strain as the template,the NP pro-tein was cloned into the eukaryotic expression vector pCMV-C-Flag through PCR amplification,the eukaryotic expression re-combinant plasmid pCMV-Flag-NP was constructed,and the expression was further verified.The protein expression levels of pCMV-Flag-NP and pCMV-HA-TRIM25 were detected at various time periods.The interaction between NP protein and TRIM25 protein was verified by co-immunoprecipitation.The co-localization of NP protein and TRIM25 protein in cells was ob-served with laser confocal microscopy.Bioinformatics analysis revealed that the NP protein consists of 498 amino acids and 20 amino acids,and is an unstable hydrophilic protein.The NP protein has multiple phosphorylation sites,as well as N-glycosyla-tion and O-glycosylation sites,but no transmembrane domain or signal peptide domain.Additionally,the NP protein's second-ary structure consists of a high proportion of alpha-helices and random coils.The molecular docking prediction results indicated that IAV NP interacts with TRIM25 protein and has multiple potential interaction sites,including the 233rd alanine,234th ala-nine,236th lysine,and 440th alanine of the NP protein.After successfully constructing and expressing the IAV NP protein,we verified the interaction between IAV NP and TRIM25 protein by immunoprecipitation and laser confocal microscopy obser-vations.Our results together suggested that the structure of the IAV NP protein is closely related to its function,and its im-portance to the virus is clear.In addition,the interaction between IAV NP and TRIM25 protein may be associated with TRIM25's anti-influenza virus mechanism.Further in-depth research may provide new ideas for anti-influenza virus strategies.

This study was aimed at establishing a method of ultra-fast quantitative PCR for Brucella detection.We used an exogenous recombinant plasmid as the internal reference and targeted the T4SS secretion system,an important Brucella viru-lence factor,to design specific primers and probes.The sensitivity,specificity,and repeatability of this method were evaluated,and a standard curve was constructed.The coincidence rate of detection findings with this method versus quantitative PCR was determined.This method markedly decreased the detection time to only 10 minutes.The standard curve demonstrated a good linear relationship(Y=-3.410 7x+38.357,R2=0.998 5)with a low minimum detection limit of 10 copies/μL.The method exhibited good specificity and did not specifically amplify several common clinical bacteria other than Brucella.The de-tection of three concentrations of positive plasmids yielded coefficients of variation(CVs)of 0.20％to 0.91％,thus demonstra-ting the method's excellent repeatability.Furthermore,140 clinical samples were analyzed concurrently with the fluorescence PCR method,which yielded a 100％compliance rate and consistent results.Our findings indicated that the Brucella ultra-fast quantitative PCR was ultrafast;had high sensitivity,high specificity,and good specificity;and can be used for the clinical de-tection of Brucella and emergency investigation of epidemics.Therefore,this method is valuable for the early diagnosis of Bru-cella.

The Animal Molecular Biology course is a crucial and foundational course for both Animal Medicine and Animal Science majors.Apart from teaching fundamental principles of molecular biology,the course provides updated applications of these principles in the field of animal science research.Im-portantly,it plays a fundamental role in cultivating students'research capabilities.With the rise of over-whelming information and their optimal utilization,the demand for integrating digital education with tradi-tional teaching methods is increasing.Based on the five years of teaching practice,this paper summarizes four highlights of the course:the construction of teaching resource,the restructuring of teaching syllabus,the adjustment of classroom teaching hours,and the improvement of assessment methodology.It focuses on Electronic-Learning"E(E-Learning)",offline classroom intensive teaching"C(Classroom)",and post-class extension to construct a blended teaching model that integrates Electronic-Learning and Class-room teaching,namely the"E+C"blended teaching model.Offline classroom teaching emphasizes the combination of theory and knowledge systems,while online Electronic-Learning mainly focuses on popular science and interesting aspects to stimulate students' interests and enthusiasm in learning.Over five years of practice,the"E+C"blended model has been proven to exert a good teaching effect.Students have reported significant gains from the course,with tightly connected and strongly complementary class-room teaching and E-Learning,which greatly aids in mastering professional knowledge.It also cultivates intrinsic motivation for learning and enhances the sense of accomplishment in acquiring knowledge,sig-nificantly improving teaching effectiveness.