AIM:To establish an optimal protocol for isolating and extracting mouse hepatocellular carcinoma H22 cell-detrived microparticles(H22-MPs),evaluate their stimulating effect on dendritic cells(DC),and investigate the cytotoxic capacity of CD8+T cells induced by H22-MPs on hepatocellular carcinoma cells.METHODS:The H22-MPs were extracted by high-speed differential centrifugation,and the morphology of H22-MPs was observed by transmis-sion electron microscopy.The particle size distribution of H22-MPs was detected by dynamic light scattering,and the ex-pression of vesicle surface markers was detected by Western blot.The protein components and pathways involved by MPs were analyzed by labeling free quantitative proteomics combining with bioinformatics GO and KEGG enrichment methods.The effect of MPs antigen on the expression of CD11c,a mature marker of DC,was detected by laser confocal microscopy.Next,CD8+T cells were sorted by magnetic beads,and the secretion of tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ)and interleukin-2(IL-2)in the supernatant of CD8+T cells stimulated by DC loaded with H22-MPs was detected by ELISA.Lactate dehydrogenase(LDH)test was conducted to evaluate the specific killing effect of CD8+T cells induced by DC loaded with H22-MPs on hepatocellular carcinoma cells.RESULTS:The results showed that the morphology,size and vesicle markers of H22-MPs met the requirements.Most of the H22-MPs protein antigens were derived from its parent cell H22 and coordinated various signal transduction pathways in the cells.Subsequently,it was detected that H22-MPs increased the expression of CD11c on the surface of DC,and H22-MPs were mainly localized in the lysosomes of DC.In addition,DC loaded with H22-MPs stimulated CD8+T to release high levels of TNF-α,IFN-γ and IL-2,and it also pro-moted CD8+T to overexpress CD69,thus inducing CD8+T to killhepatocellular carcinoma cells specifically.CONCLU-SION:H22-MPs promote the maturation of DC,and the mature DC may induce CD8+T cells to play an anti-hepatocellu-lar carcinoma immune response by promoting the activation of CD8+T cells.

Objective:To investigate the effects of target-oriented liquid therapy on the treatment outcomes of skin grafting in patients with severe burns.Methods:A total of 60 patients with severe burns (total burn area over 20%, and deep second-degree burn area ≥ 10%) who were scheduled for skin graft surgery at the Marine Police Corps Hospital of Chinese People's Armed Police Force from January 2019 to January 2022 were included in this cohort study. The patients were randomly divided into a control group (conventional fluid therapy) and an observation group (target-oriented fluid therapy) using a random number table method, with 30 patients in each group.The levels of lactate (Lac) and high-sensitivity cardiac troponin I (cTnI) were compared between the two groups. The usage of colloid and crystalloid fluids, urine output, length of hospital stay, and the incidence of complications were compared between the two groups.Results:At the end of the surgery, the levels of Lac and cTnI in the observation group were (1.13 ± 0.22) mmol/L and (0.95 ± 0.25) μg/L, respectively, both of which were significantly lower than those in the control group [(1.52 ± 0.34) mmol/L, (1.10 ± 0.31) μg/L; t = 5.24, 0.83, P < 0.001, P = 0.044]. The differences in Lac and cTnI levels in the observation group were (0.53 ± 0.36) and (0.13 ± 0.07), respectively, which were significantly greater than those in the control group [(0.23 ± 0.16), (0.02 ± 0.01), t = -4.17 and -3.45, P < 0.001, P = 0.001]. The volumes of colloid and crystalloid fluids and the length of hospital stay in the observation group were (960.25 ± 153.32) mL, (1 680.52 ± 253.08) mL, and (51.36 ± 17.25) days, respectively. These values were significantly lower or shorter than those in the control group [(1 459.73 ± 203.41) mL, (2 401.89 ± 301.23) mL, (81.05 ± 25.08) days, t = 10.74, 10.04, 5.34,all P < 0.001]. The incidences of postoperative infection, pneumonia, and arrhythmia in the observation group were 33.33% (10/30), 16.67% (5/30), and 16.67% (5/30), respectively, which were significantly lower than those in the control group [63.33% (19/30), 46.67% (14/30), 30.00% (15/30); χ2 = 5.41, 6.24, 7.50, P = 0.020, P = 0.013, P = 0.006]. However, there was no statistically significant difference in acute lung injury, heart failure, and myocardial injury between the two groups ( χ2 = 1.36, 1.92, 0.29; P = 0.243, 0.166, 0.592). Conclusions:Compared with traditional fluid replacement therapy, target-oriented fluid therapy has a more significant therapeutic effect on patients with severe burns. It not only reduces fluid usage and the length of hospital stay but also decreases the incidence of infection, pneumonia, and arrhythmia in these patients.

Objective To investigate the efficacy of intratumoral and peritumoral radiomics features besed on biparametric MRI combined with pathological parameters in predicting biochemical recurrence(BCR)after radical prostatectomy(RP).Methods A total of 295 prostate cancer(PCa)patients who underwent RP were retrospectively selected and divided into training set(n=206)and test set(n=89)in a 7∶3 ratio.Subgroups were stratified based on prostate specific antigen(PSA)results during 24-month postopera-tive follow-up.Radiomics features were extracted and integrated from both intratumoral and peritumoral 3 mm dual-regions on T2WI and apparent diffusion coefficient(ADC)sequences.Feature selection was performed using independent sample t-tests,maximum relevance and minimum redundancy(mRMR)algorithm,and least absolute shrinkage and selection operator(LASSO)regression.Binary logis-tic regression analysis was used to screen the independent pathological risk factors.Radiomics models(intratumoral,peritumoral,and combined)and a pathological model were constructed based on the Light gradient boosting machine(LightGBM)algorithm.A nomo-gram model was developed by integrating the pathological model with the combined model.The model efficacy was evaluated via the receiver operating characteristic(ROC)curves,and the DeLong's test and net reclassification improvement(NRI)were used to com-pare the efficacy differences among the models.Results Finally,the optimal feature numbers for the intratumoral,peritumoral,and combined models were 5,6,and 6,respectively.In the test set,the combined model demonstrated significantly higher area under the curve(AUC)than the intratumoral model(0.905 vs 0.815,P＜0.05).Ki-67,extracapsular extension,and positive surgical margin were identified as independent pathological predictors of BCR(P＜0.05).The nomogram model demonstrated significantly higher AUC than the pathological model(0.931 vs 0.786,P＜0.05),with no significant difference versus the combined model(0.931 vs 0.905,P＞0.05).Conclusion The nomogram model integrating intratumoral,peritumoral radiomics features with pathological parameters can effectively predict the occurrence of BCR within 2 years after PCa surgery.

AIM:To establish an optimal protocol for isolating and extracting mouse hepatocellular carcinoma H22 cell-detrived microparticles(H22-MPs),evaluate their stimulating effect on dendritic cells(DC),and investigate the cytotoxic capacity of CD8+T cells induced by H22-MPs on hepatocellular carcinoma cells.METHODS:The H22-MPs were extracted by high-speed differential centrifugation,and the morphology of H22-MPs was observed by transmis-sion electron microscopy.The particle size distribution of H22-MPs was detected by dynamic light scattering,and the ex-pression of vesicle surface markers was detected by Western blot.The protein components and pathways involved by MPs were analyzed by labeling free quantitative proteomics combining with bioinformatics GO and KEGG enrichment methods.The effect of MPs antigen on the expression of CD11c,a mature marker of DC,was detected by laser confocal microscopy.Next,CD8+T cells were sorted by magnetic beads,and the secretion of tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ)and interleukin-2(IL-2)in the supernatant of CD8+T cells stimulated by DC loaded with H22-MPs was detected by ELISA.Lactate dehydrogenase(LDH)test was conducted to evaluate the specific killing effect of CD8+T cells induced by DC loaded with H22-MPs on hepatocellular carcinoma cells.RESULTS:The results showed that the morphology,size and vesicle markers of H22-MPs met the requirements.Most of the H22-MPs protein antigens were derived from its parent cell H22 and coordinated various signal transduction pathways in the cells.Subsequently,it was detected that H22-MPs increased the expression of CD11c on the surface of DC,and H22-MPs were mainly localized in the lysosomes of DC.In addition,DC loaded with H22-MPs stimulated CD8+T to release high levels of TNF-α,IFN-γ and IL-2,and it also pro-moted CD8+T to overexpress CD69,thus inducing CD8+T to killhepatocellular carcinoma cells specifically.CONCLU-SION:H22-MPs promote the maturation of DC,and the mature DC may induce CD8+T cells to play an anti-hepatocellu-lar carcinoma immune response by promoting the activation of CD8+T cells.

Objective:To explore the value of ultrasound imaging characteristics combined with clinical indicators in assessing the prognosis of patients with pancreatic ductal adenocarcinoma (PDAC).Methods:A retrospective analysis was conducted for patients who underwent pancreatic contrast-enhanced ultrasound (CEUS) from September 2017 to October 2023 at Peking Union Medical College Hospital and were diagnosed with PDAC based on pathological findings. Various parameters were recorded, including CA19-9 levels, tumor size, location, morphologic features, echogenicity, presence of internal cystic components, dilatation of the main pancreatic duct, peripheral vascular invasion, CEUS characteristics, presence or absence of liver metastasis, and treatment methods. In April 2024, patient survival information was obtained through telephone follow-up or review of medical records. Based on the results of the cox regression model analysis, a nomogram model of the risk of death was developed. The receiver operating characteristic (ROC) curves were applied to evaluate the predictive efficacy of the model. The calibration curves were plotted to evaluate the accuracy of the model, and clinical decision curves were used to evaluate the clinical benefit of the model.Results:This study included a total of 207 patients with PDAC. As of April 2024, 71 patients were alive and 136 died, with a median survival time of 14 months (95% CI: 12 -17). Multivariate analysis confirmed that the elevated CA19-9 ( HR=1.689, 95% CI: 1.102-2.588), tumor size >4 cm ( HR=1.641, 95% CI: 1.159-2.322), taller-than-wide shapes ( HR=1.450, 95% CI: 1.019-2.065), incomplete hypo-enhancement ( HR=1.618, 95% CI: 1.100-2.380), and liver metastasis ( HR=1.687, 95% CI: 1.175-2.423) were independent risk factors for survival in patients with PDAC. A nomogram model was further constructed for 6-month, 12-month and 3-year survival of patients with PDAC. The areas under the ROC curve were 0.679, 0.705 and 0.815, respectively. The calibration curves suggested that the model was more accurate, and the clinical decision curves showed that the model had a better clinical benefit. Conclusion:The combined use of ultrasound imaging characteristics and clinical indicators could effectively predict the prognosis of PDAC patients. Specifically, tumor size >4 cm, taller-than-wide shapes, incomplete hypo-enhancement, elevated CA19-9, and the presence of liver metastasis are correlated with poorer survival outcomes. The nomogram model constructed on the basis of these factors can be used to assess the survival of patients with PDAC.

Objective:To investigate the clinical value of Delphian lymph node (DLN) metastasis (DLNM) in predicting regional lymph node involvement in patients with intrathyroidal papillary thyroid carcinoma (PTC).Methods:Clinical and pathological data from 345 consecutive patients with pathologically confirmed DLN status, who underwent initial surgical treatment at the Department of Surgical Oncology, First Affiliated Hospital of Zhejiang University School of Medicine between Jan 2020 and Dec 2022, were retrospectively analyzed.Results:DLNM was identified in 61 patients (17.7%). Univariate analysis revealed significant associations between DLNM and male sex, elevated preoperative thyroglobulin levels, larger tumor size, maximum tumor diameter >10 mm, bilateral lesions, multifocality, lymphovascular invasion, and lymph nodes metastases in pretracheal, paratracheal, and lateral cervical(all P ≤0.001). Elevated thyroglobulin antibody levels ( χ2=6.201, P=0.013) and Hashimoto's thyroiditis ( χ2=11.340, P<0.001) were protective factors for DLNM. Multivariate analysis identified male sex, lymphovascular invasion, pretracheal, and paratracheal lymph node metastases ( χ2=6.689, P=0.010; χ2=8.163, P=0.004; χ2=7.605, P=0.006; χ2=8.324, P=0.004) as independent risk indicators for DLNM. Patients with DLNM exhibited significantly higher risks of lymph nodes metastases in pretracheal ( χ2=27.307, P<0.001), paratracheal ( χ2=38.697, P<0.001), and lateral cervical ( χ2=36.459, P<0.001). Conclusion:DLNM demonstrates predictive value for both central compartment and lateral cervical lymph node metastases, warranting particular attention to meticulous dissection of the prelaryngeal region during surgery.

Objective To investigate the efficacy of intratumoral and peritumoral radiomics features besed on biparametric MRI combined with pathological parameters in predicting biochemical recurrence(BCR)after radical prostatectomy(RP).Methods A total of 295 prostate cancer(PCa)patients who underwent RP were retrospectively selected and divided into training set(n=206)and test set(n=89)in a 7∶3 ratio.Subgroups were stratified based on prostate specific antigen(PSA)results during 24-month postopera-tive follow-up.Radiomics features were extracted and integrated from both intratumoral and peritumoral 3 mm dual-regions on T2WI and apparent diffusion coefficient(ADC)sequences.Feature selection was performed using independent sample t-tests,maximum relevance and minimum redundancy(mRMR)algorithm,and least absolute shrinkage and selection operator(LASSO)regression.Binary logis-tic regression analysis was used to screen the independent pathological risk factors.Radiomics models(intratumoral,peritumoral,and combined)and a pathological model were constructed based on the Light gradient boosting machine(LightGBM)algorithm.A nomo-gram model was developed by integrating the pathological model with the combined model.The model efficacy was evaluated via the receiver operating characteristic(ROC)curves,and the DeLong's test and net reclassification improvement(NRI)were used to com-pare the efficacy differences among the models.Results Finally,the optimal feature numbers for the intratumoral,peritumoral,and combined models were 5,6,and 6,respectively.In the test set,the combined model demonstrated significantly higher area under the curve(AUC)than the intratumoral model(0.905 vs 0.815,P＜0.05).Ki-67,extracapsular extension,and positive surgical margin were identified as independent pathological predictors of BCR(P＜0.05).The nomogram model demonstrated significantly higher AUC than the pathological model(0.931 vs 0.786,P＜0.05),with no significant difference versus the combined model(0.931 vs 0.905,P＞0.05).Conclusion The nomogram model integrating intratumoral,peritumoral radiomics features with pathological parameters can effectively predict the occurrence of BCR within 2 years after PCa surgery.

BACKGROUND:Acute exercise tends to cause skeletal muscle tissue damage and lipid metabolism disorders in vivo,but the mechanism by which acute exercise combined with electroacupuncture modulates metabolic and autophagic pathways in vivo is unclear. OBJECTIVE:To observe the changes in metabolic enzymes and autophagy levels in skeletal muscle of rats subjected to acute exercise by electroacupuncture at the acupoints of"Zusanli"and"Huantiao." METHODS:Fifty male Sprague-Dawley rats were randomly divided into three groups:quiet control group(n=10),model group(n=20),and reverse electroacupuncture group(n=20).The latter two groups were set up with two time points,i.e.immediate and 3 hours after exercise groups(n=10 per time point).The model group and the reverse electroacupuncture group underwent acute exercise training after adaptive treadmill training.The rats in the reverse electroacupuncture group underwent electroacupuncture treatment(parameters:electroacupuncture on both sides of the rats at the acupoints of"Zusanli"and"Huantiao,"continuous wave,frequency of 2 Hz,intensity of 2 mA,leaving the needle in the body for 30 minutes,once a day for 7 consecutive days)before treadmill training.Bilateral gastrocnemius muscle tissues were taken under anesthesia immediately after exercise and 3 hours after exercise,and hematoxylin-eosin staining was used to observe the histopathological changes of rat skeletal muscle.ELISA kit was used to detect the activities of hepatic lipase,fatty acid synthase,hormone-sensitive lipase,and carnitine palmitoyltransferase 1 in rat skeletal muscle tissues.Immunohistochemistry and western blot were used to detect the changes in the expression of autophagy genes. RESULTS AND CONCLUSION:After hematoxylin-eosin staining,the arrangement of gastrocnemius muscle fibers in the model group was disturbed,swollen and ruptured immediately after exercise and 3 hours after exercise.In the reverse electroacupuncture group,gastrocnemius muscle fibers were tightly arranged and the number of swollen and ruptured cells was greatly reduced immediately after exercise and 3 hours after exercise,and there was no significant difference when compared with the quiet control group.Compared with the quiet control group,the activities of hepatic lipase and fatty acid synthase were lower while the activities of lipoprotein lipase,hormone-sensitive lipase,and carnitine palmitoyltransferase 1 were higher in the model group and the reverse electroacupuncture group 3 hours after exercise(P＜0.05 or P＜0.01).Compared with the model group,the activities of lipoprotein lipase and carnitine palmitoyltransferase 1 were higher in the reverse electroacupuncture group immediately after exercise(P＜0.05),while the activity of lipoprotein lipase was higher and the activity of hormone-sensitive lipase was lower in the reverse electroacupuncture group 3 hours after exercise(P＜0.01).Immunohistochemical results showed that compared with the quiet control group,the expression of P62,autophagy-related gene 5 and autophagy-related gene 7 was higher in the model group immediately and 3 hours after exercise,as well as in the reverse electroacupuncture group immediately after exercise(P＜0.05 or P＜0.01);compared with the model group,the expression of P62 and autophagy-related gene 7 was lower in the reverse electroacupuncture group immediately and 3 hours after exercise(P＜0.05).Western blot results showed that the protein expression of P62 and autophagy-related gene 7 in the reverse electroacupuncture group was lower than that in the model group immediately after exercise(P＜0.05);the protein expression of Parkin in the model group was higher than that in the quiet control group immediately and 3 hours after exercise(P＜0.05);and the protein expression of Parkin in the reverse electroacupuncture group was lower than that in the model group immediately and 3 hours after exercise(P＜0.05).To conclude,acute exercise induces disorders,swelling and rupture of gastrocnemius muscle fibers in rats and electroacupuncture on both sides of the acupoints of"Zusanli"and"Huantiao"can improve the level of lipid metabolism and regulate autophagy cells in rat skeletal muscle,preventing the disorders of lipid metabolism and damage of gastrocnemius muscle tissues caused by acute exercise.The mechanism may be closely related to the regulation of autophagy-related factor P62,autophagy-related gene 5,autophagy-related gene 7,and Parkin protein expression to promote the occurrence of autophagy or regulate the autophagy pathway in rat skeletal muscle cells.

Obesity, as a chronic recurrent disease, has become a major public health challenge in China. To implement the requirements of the Healthy China Initiative (2019—2030), under domestic guidelines or consensus statements on overweight and obesity, and in alignment with the latest scientific advances globally, the Quality control protocol for adult overweight and obesity screening in health management (examination) institutions (2025 edition) was developed. This protocol was drafted by the Health Management Center of Shanghai Changzheng Hospital and formulated through multiple rounds of deliberation by experts in China’s health examination quality control field. The protocol establishes unified standards for screening facilities, personnel qualifications, and measurement or testing procedures. It defines specific screening items, outlines a standardized screening pathway, and sets requirements for the final medical review, ensuring the scientific validity, effectiveness, and safety of the screening process. The implementation of this protocol will enhance the consistency of weight management practices for adults across health examination institutions and strengthen the quality control of overweight and obesity screening programs.

Objective: To investigate the effects of polydatin on a high-fat diet-induced non-alcoholic fatty liver disease(NAFLD) mouse model and hepatoma G2(HepG2) cell model, and to reveal its potential molecular mechanisms. Methods: Thirty 6-week-old male SPF C57BL/6J mice were randomly divided into a normal diet group and a high-fat diet group. After the NAFLD mouse model was established in the high-fat diet group, they were further divided into a model group and a polydatin treatment group. The polydatin treatment group was administered polydatin by gavage at a dose of 250 mg/(kg·d) for 10 weeks, during which body weight was monitored and oral glucose and insulin tolerance tests were performed. At the end of the experiment, a series of tests to evaluate the effects of polydatin on mouse liver weight, blood lipids, liver lipid accumulation, and liver injury markers were performed. The expression of G0/G1 switch gene 2(G0S2) and adipose triglyceride lipase(ATGL) was measured by qRT-PCR and Western blot, and gene expression was further verified using immunohistochemical staining. The effects of polydatin on HepG2 cell activity was assessed by CCK-8 assay, lipid accumulation was observed by oil red O staining, and the expression of G0S2 and ATGL was detected by qRT-PCR and Western blot. Results: Polydatin significantly reduced the body weight, liver weight, and serum and liver tissue levels of aspartate aminotransferase(AST), alanine aminotransferase(ALT), triglyceride(TG), and total cholesterol (TC) in mice (P < 0. 05) , al⁃leviated pathological liver damage , decreased G0S2 expression (P < 0. 05) , and increased ATGL expression (P <0. 05) . At the cellular level , polydatin reduced lipid droplet accumulation , improved lipid metabolism , decreased G0S2 expression ( P < 0. 05 ) , and increased ATGL expression ( P < 0. 05 ) . Even in cells with knockdown of G0S2 , polydatin still promoted fat decomposition (P < 0. 01) . Conclusion Polydatin promotes hepatic fat break⁃down by regulating the expression of G0S2 and ATGL , helping to alleviate metabolic disorders and liver damage in the NAFLD mouse model caused by a high⁃fat diet , offering a new strategy for treating NAFLD.