ObjectiveTo investigate the therapeutic mechanism of Xuefu Zhuyutang （XFZYT） for metabolic-associated fatty liver disease （MAFLD） through integrated network pharmacology and animal experiments. MethodsNetwork pharmacology was utilized to predict the core components， key therapeutic targets， and signaling pathways of XFZYT in the treatment of MAFLD. For animal experiments， a rat model of MAFLD was established by feeding a high-cholesterol diet for 4 weeks. Intervention was then administered with low-dose （2 g·kg^-1） and high-dose （4 g·kg^-1） XFZYT for 2 weeks. Biochemical assays were performed to measure the serum levels of aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， total cholesterol （TC）， triglycerides （TG）， high-density lipoprotein （HDL）， and low-density lipoprotein （LDL）. In addition， the activities of superoxide dismutase （SOD） and catalase （CAT） and levels of malondialdehyde （MDA） and glutathione （GSH） in the serum were measured. The same way was adopted to measure the levels of TC and TG in the liver tissue. Enzyme-linked immunosorbent assay （ELISA） was employed to quantify the serum levels of interleukin （IL）-6， IL-1β， and tumor necrosis factor-alpha （TNF-α）. Histopathological evaluations included hematoxylin and eosin （HE） staining for liver tissue morphology， Oil Red O staining for lipid deposition， and dihydroethidium （DHE） probe staining for reactive oxygen species （ROS） levels. Western blot analysis was conducted to assess the protein levels of AMP-activated protein kinase （AMPK）， phosphorylated （p）-AMPK， nuclear factor erythroid 2-related factor 2 （Nrf2）， heme oxygenase-1 （HO-1）， nuclear factor-kappa B （NF-κB）， and p-NF-κB in the liver tissue. Untargeted metabolomics analysis of the serum was performed by liquid chromatography-tandem mass spectrometry （LC-MS/MS）. ResultsNetwork pharmacology analysis predicted 155 potential targets of XFZYT for MAFLD treatment， with core targets including signal transducer and activator of transcription 3 （STAT3）， protein kinase B1 （Akt1）， TNF， and IL-6. Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment primarily implicated the AMPK signaling pathway. Animal experiments demonstrated that compared with the normal group， the model group exhibited dyslipidemia， hepatic function impairment， pronounced hepatic lipid deposition， and inflammatory manifestations， with elevated serum levels of AST， ALT， TC， TG， LDL， and MDA （P<0.05）， reduced HDL and GSH levels plus decreased SOD and CAT activities （P<0.05）， downregulated protein levels of Nrf2， HO-1， and p-AMPK （P<0.05）， and upregulated protein level of p-NF-κB （P<0.05） in the liver tissue. Compared with the model group， XFZYT intervention groups showed significant amelioration of dyslipidemia and hepatic function impairment， markedly reduced hepatic lipid deposition and inflammatory cell infiltration， decreased serum levels of AST， ALT， TC， TG， LDL， and MDA （P<0.05）， increased HDL and GSH levels plus enhanced SOD and CAT activities （P<0.05）， upregulated protein levels of Nrf2， HO-1， and p-AMPK （P<0.05）， and downregulated protein level of p-NF-κB （P<0.05）. Serum metabolomics revealed 511 differentially expressed metabolites （231 upregulated and 280 downregulated） between normal and model groups， while XFZYT groups versus model group showed 94 differential metabolites （51 upregulated and 43 downregulated）. Among them， 11 metabolites displayed the most significant alterations， with enriched pathways including glycerolipid metabolism， cholesterol metabolism， and insulin resistance， multiple of which demonstrated AMPK association. ConclusionXFZYT alleviates MAFLD by regulating the AMPK signaling pathway and associated metabolic networks.

Bronchiolar adenoma (BA) is a rare type of lung tumor characterized by bilayered epithelial cells having a continuous basal layer and a luminal layer. It resembles mucinous adenocarcinoma (MA) on frozen section, with difficulty in distinguishing the basal layer. Immunohistochemistry is the best choice for verifying the diagnosis. This study aimed to comprehensively characterize three cases of BA-combined carcinoma using clinical, histopathological, and genetic features. BA and carcinoma sections were subjected to next-generation sequencing, respectively. It was hypothesized that while different mutation forms matched different regions, BA and lung adenocarcinoma shared the same gene mutation when they co-occurred in the same location. BA with extensive carcinoma is extremely rare and presents diagnostic challenges due to its overlap with conditions such as MA. Because of its distinctive morphological characteristics, BA may be regarded as a low-grade malignancy, particularly during a confusing evaluation. A multifaceted examination of clinical, radiological, immunohistochemical, and genetic data is necessary for an accurate diagnosis.

Abstract To clarify the service targets, service contents, and safeguard mechanisms of the basic public service program for student vision health management, the study grounded in the current status of child and adolescent myopia prevention and control in China and integrated with the practical experience and innovative practices of Wuhan National Demonstration Area for Student Vision Health Management, takes education as the main thread and schools as the base, consolidating resources from multiple stakeholders including education administrative departments, schools, families, and professional technical service institutions. Centering on four core components—vision health education, monitoring and record keeping, risk prevention and control, and dynamic management, the study constructs a multi tiered basic public service system for vision health management of primary and secondary school students. It further proposes the use of an intelligent management platform and a "three tier color coded early warning" mechanism (red, yellow, blue) to prevent and control student myopia at the source, providing schools with replicable and operable implementation pathways to enhance the level of student vision health management.

Objective: To examine the effects of combined aerobic and resistance exercise on health related physical fitness (HPF) and depressive symptoms among senior high school freshmen, and to examine whether HPF mediates the relationship between exercise intervention and depressive symptoms, so as to provide a scientific basis for the coordinated promotion of mental health and physical fitness in schools. Methods: In March 2022, 190 senior high school freshmen from a secondary school of Suzhou in Anhui Province were selected by the random cluster sampling method and randomly assigned to an intervention group ( n = 99) and a control group ( n =91). The control group maintained routine activities, while the intervention group underwent a 10 week combined aerobic and resistance exercise program, three sessions per week, 40 minutes per session. Height, weight, standing long jump, sit ups, 20 m shuttle run, and sit and reach were measured before and after the intervention. Depressive symptoms were assessed using the Center for Epidemiological Studies Depression Scale (CES-D). Analysis of covariance (ANCOVA), Mann-Whitney U test, and Wilcoxon test were employed to compare outcomes before and after the intervention, and mediation analysis was conducted. Results: After controlling for pre test scores, ANCOVA results showed that compared with the control group, the intervention group demonstrated the following post intervention changes: a lower body mass index of 0.1 kg/m 2 among girls; higher standing long jump (13.2 cm for boys, 7.4 cm for girls), higher sit ups (5.5 reps for boys, 3.7 reps for girls), higher 20 m shuttle run (7.4 times for boys, 5.1 times for girls), and higher sit and reach (3.2 cm for boys, 0.4 cm for girls) ( F =17.88-84.44); and lower CES-D scores (5.5 for boys, 4.0 for girls) ( Z =-2.51, -2.06) (all P <0.05). The indirect effect of HPF in the relationship between combined aerobic and resistance exercise and depressive symptoms was -2.11, supporting a partial mediation effect ( P < 0.01 ). Conclusions Combined aerobic and resistance exercise improves HPF and alleviates depressive symptoms to varying degrees among senior high school freshmen. HPF served as a partial mediator through which exercise intervention reduces depressive symptoms.

Objective To analyze the application effect of narrow band imaging(NBI)combined with magnifying endoscopy(ME)in early gastric cancer(EGC)screening.Methods A total of 199 patients who found focal lesions by ordinary white light endoscopy(WLE)were selected from April 2022 to October 2023.NBI combined with ME examination was performed,and histopathological examination of the specimen obtained from the lesion area was conducted.The pathological diagnosis results were recorded,and the endoscopic image qualities of WLE and NBI combined with ME examination were observed.The consistency between WLE,NBI combined with ME and patho-logical diagnosis was analyzed.The diagnostic value of WLE and NBI combined with ME for EGC was evaluated.Results The image qualities of microvascular and glandular duct by NBI combined with ME were better than those by WLE(P<0.05).The consistency between WLE diagnosis and pathological diagnosis was comparatively strong(Kappa=0.687).The consistency between NBI combined with ME and pathological diagnosis was very strong(Kappa=0.947).The consistency between NBI combined with ME and pathological diagnosis was better than that of WLE.The sensitivity,specificity,positive predictive value,negative predictive value,accuracy,and area under the curve(AUC)of NBI combined with ME in diagnosing EGC were higher than those of WLE,with statistically significant differences(P<0.05).The AUC of NBI combined with ME for diagnosing EGC was higher than that of WLE,with statistically significant difference(Z=3.493,P=0.000).Conclusion NBI combined with ME can increase the diagnostic efficiency of EGC,along with good image quality.

Objective To explore the application value of wide awake local anesthesia no tourniquet(WALANT)technique in outpatient surgery for locking of metacarpophalangeal joint with extension lag.Methods The clinical data of 6 patients with locking of meta-carpophalangeal joint with extension lag in Danyang People's Hospital from January 2019 to October 2023 were retrospectively analyzed.The patients were received outpatient surgery under the WALLANT technique for release,and lidocaine mixed solution containing 1∶100 000 epinephrine was injected into the proximal midpoint of the volar projection of the metacarpophalangeal joint.The joint was exposed with a volar or dorsal finger web incision to determine the unrestricted structure,and the collateral ligament and paralateral collateral ligament with high tension were cut off.The intraoperative blood loss,postoperative incision healing and complications were recorded.Visual analogue scale(VAS)was used to evaluate the intraoperative pain,and the range of motion of metacarpophalangeal joint and total active movement(TAM)of finger joint were observed during postoperative follow-up.Results The incision of patients were healed successfully in the first phase after surgery,without wound necrosis.The anesthesia effects of patients were all satisfied and the operation was successfully completed.The VAS score was less than 3 points and there was only a small amount of bleeding during the operation.The recovery of joint flexion and extension movements could be observed during the operation,and the TAM score after the operation was 20 points.No significant change was found on the range of motion of metacarpophalangeal joint or TAM of finger joint between the injured finger and the corresponding healthy finger(P>0.05).Conclusion WALANT technique for locking of metacarpophalangeal joint with extension lag surgery has good anesthesia effect,less bleeding in the incision,and clear vision of the surgery.It can avoid vascular and nerve injuries,observe the recovery of joint activities during the operation and relieve pain of patients,which is conducive to outpatient surgery and saving medical and social resources at the same time.

Hepatocellular carcinoma(HCC),which is essentially primary liver cancer,is closely related to CD8+T cell immune infiltration and immune suppression.We constructed a CD8+T cells related risk score model to pre-dict the prognosis of HCC patients and provided therapeutic guidance based on the risk score.Using integrated bulk RNA sequencing(RNA-seq)and single-cell RNA sequencing(scRNA-seq)datasets,we identified stable CD8+T cell signatures.Based on these signatures,a 3-gene risk score model,comprised of KLRB1,RGS2,and TN-FRSF1B was constructed.The risk score model was well validated through an independent external validation co-hort.We divided patients into high-risk and low-risk groups according to the risk score and compared the differ-ences in immune microenvironment between these two groups.Compared with low-risk patients,high-risk patients have higher M2-type macrophage content(P＜0.0001)and lower CD8+T cells infiltration(P＜0.0001).High-risk patients predict worse response to immunotherapy treatment than low-risk patients(P＜0.01).Drug sensitivity a-nalysis shows that PI3K-β inhibitor AZD6482 and TGFβRII inhibitor SB505124 may be suitable therapies for high-risk patients,while the IGF-1R inhibitor BMS-754807 or the novel pyrimidine-based anti-tumor metabolic drug Gemcitabine could be potential therapeutic choices for low-risk patients.Moreover,expression of these 3-gene mod-el was verified by immunohistochemistry.In summary,the establishment and validation of a CD8+T cell-derived risk model can more accurately predict the prognosis of HCC patients and guide the construction of personalized treatment plans.

AIM To establish a UPLC method for the simultaneous content determination of liquiritin,ammonium glycyrrhizate,naringin,neohesperidin,hesperidin,rosmarinic acid,baicalin,wogonoside,baicalein,wogonin and praeruptorin A in Tongxuan Lifei Pills,and to make chemometric investigation.METHODS The analysis was performed on a 30 ℃ thermostatic SVEA C18 column(2.1 mm×150 mm,2.5 μm),with the mobile phase comprising of acetonitrile-0.1％phosphoric acid flowing at 0.5 mL/min in a gradient elution manner,and the detection wavelength was set at 250 nm.Subsequently,cluster analysis,principal component analysis and partial least square discriminant analysis were performed.RESULTS Eleven constituents showed good linear relationships within their own ranges(R2＞0.990 0),whose average recoveries were 90.00％-98.32％with the RSDs of 0.35％-1.89％.Forty batches of samples were clustered into 3 types.Baicalein,baicalin,liquiritin,wogonin,wogonoside and neohesperidin were taken as quality differential markers.CONCLUSION This simple and reproducible method can provide the basis for quality control and evaluation of Tongxuan Lifei Pills.

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.