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.

BACKGROUND:The repair process of myocardial injury involves complex cellular and molecular mechanisms,especially mitochondrial calcium homeostasis,macrophage autophagy and pyroptosis pathways.Traditional Chinese medicine(TCM)has shown significant clinical efficacy in improving myocardial injury,but its mechanism of action needs to be thoroughly investigated. OBJECTIVE:To investigate the role of mitochondrial calcium homeostasis-mediated macrophage autophagy and pyroptosis pathways in myocardial injury,and to summarize the progress of TCM in this field. METHODS:A computerized search was performed for relevant literature from the database inception to March 2024 in the Web of Science,PubMed and CNKI.The search terms were"mitochondrial calcium homeostasis,macrophage autophagy,macrophage pyroptosis,traditional Chinese medicine,myocardial injury,myocardial injury reperfusion"in Chinese and English.Through literature review,we analyzed the relationship between mitochondrial calcium homeostasis and macrophage autophagy and pyroptosis,explored the mechanism of their roles in myocardial injury,and summarized the pathways of multi-targeted,multi-pathway effects of TCM. RESULTS AND CONCLUSION:The maintenance of mitochondrial calcium homeostasis has been found to be closely related to the normal function of cardiomyocytes.Macrophages can participate in the repair process of myocardial injury through autophagy and pyroptosis pathways.Autophagy contributes to cell clearance and regulation of inflammatory response,while pyroptosis affects myocardial repair by releasing inflammatory factors.TCM regulates mitochondrial calcium homeostasis and macrophage function through multiple mechanisms.For example,astragalosid regulates calcium homeostasis by lowering mitochondrial membrane potential and inhibiting cytochrome C,and epimedium glycoside plays a role in reducing β-amyloid deposition.In addition,herbal compounds and single drugs promote myocardial repair by activating or inhibiting specific signaling pathways,such as PI3K/AKT and nuclear factor-κB signaling pathways.Future studies should focus on the interactions between mitochondrial calcium homeostasis,autophagy and pyroptosis pathways,as well as how TCM can exert therapeutic effects through these pathways to provide new strategies and drugs for the treatment of myocardial injury.

This study established an indirect ELISA method for antibody detection based on human rotavirus group A(RVA)VP6 protein,to provide technical support for RVA antibody detection.An ELISA method for detecting RVA antibodies was established through a checkerboard assay for optimization of the reaction conditions with VP6 protein of RVA as the diagnostic antigen.Serum samples from healthy children and patients infected with RVA were collected and detected with the ELISA method.The optimal work-ing conditions for ELISA involved coating the ELISA plate with VP6 protein at a concentration of 2 μg/mL and diluting the serum at a ratio of 1∶250.The critical value of negative and positive samples was 0.137.The ELISA method had good specificity,sensitivity,and repeatability,and showed a 95%consistency rate with the western blotting antibody detection method.We tested 370 serum samples collected from children,which showed an antibody positivity rate of 81.1%.We additionally tested the acute and recovery phase sera from 15 patients infected with RVA,and observed a significantly higher RVA antibody titer in the recovery phase serum than the acute infection phase serum.The ELISA antibody detection method showed good specificity,sensitivity,and reproducibility,and therefore can be used for RVA antibody detection and auxiliary diagnosis of RVA infection.

This study expressed the VP6 protein of human group A rotavirus in a prokaryotic expression system and prepared mouse polyclonal antibodies to this protein.The human rotavirus VP6 gene was amplified through RT-PCR from fecal samples infected with group A rotavirus and cloned into the pET-32a vector to construct the recombinant pET-32a-VP6 prokaryotic expression plas-mid.The recombinant plasmid was transformed into Escherichia coli BL21(DE3).VP6 protein was expressed through induction with isopropyl-β-D-thiopyranoside(IPTG),purified with nickel nitrilotriacetic acid(Ni-NTA)affinity chromatography,and identified through SDS-PAGE and western blotting.Polyclonal antibodies to the VP6 protein were obtained by immunization of BALB/c mice with VP6 protein.Antibody titers were determined through indirect ELISA.The recombinant expression plasmid pET-32a-VP6 was con-structed,and VP6 protein was expressed primarily in inclusion bodies.Polyclonal antibodies to VP6 protein with a titer of 1∶32 000 were obtained by immunization of mice with VP6 protein purified by affinity chromatography.The antibodies specifically reacted with VP6 protein.The VP6 protein of human group A rotavirus was expressed and purified,and mouse polyclonal antibodies to this protein were prepared,thus providing a foundation for the preparation of a diagnostic antigen for human group A rotavirus and the establishment of serological detection methods.

Objective To explore the impact of the number of negative lymph node resections on the overall survival and recurrence rate of patients with ovarian cancer.Methods A retrospective selection was made of 150 ovarian cancer patients admitted to our hospital from October 2021 to October 2023 as the research subjects.According to the standard of the number of negative lymph nodes removed,they were divided into three groups:T1 group(≤17),T2 group(18～27),and T3 group(≥28).Clinical characteristics and surgical indicators were collected,and the recurrence and survival situations were analyzed.Results The clinical characteristic analysis results showed that there were no statistically significant differences among the three groups in terms of age,meno-pausal status,BMI,CCI,family history of tumors,etc.(P>0.05).There were no significant differences in surgi-cal methods,residual lesion size,postoperative complications among the three groups(P>0.05).In terms of operation time,T1 group0.05),but with the increase in the number of lymph node resections(from T1 group to T3 group),the recurrence rate showed a statistically significant downward trend(P<0.05).Cox regression analysis showed that the univariate analysis indicated that for each additional negative lymph node,the risk of death decreased by 4.7%(HR=0.953,P<0.05);the multivariate analysis(after adjusting for confounding factors)showed that the risk decreased by 5.8%(HR=0.942,P<0.05).Restricted cubic spline(RCS)analysis confirmed a significant linear association between the two(P<0.05).With 5 lymph nodes as the reference,the risk ratios for 10,20,and 30 resections were 1.003,0.760,and 0.317,respectively,and the trend showed that the risk of death contin-ued to decrease as the number of lymph nodes increased.Multivariate Cox analysis showed that the risk of death in T3 group was 68%lower than that in T1 group(P<0.05),the risk in FIGO stage Ⅲ was 3.11 times higher(P<0.05),and the risk in high-grade tumors was 2.80 times higher(P<0.05).Conclusions The number of nega-tive lymph nodes removed in patients with ovarian cancer is linearly correlated with prognosis.The more lymph nodes removed,the lower the risk of death for the patients.Adequate lymph node removal should be an important part of the surgical treatment for ovarian cancer.

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 explore the clinical efficacy of retrograde tibial intramedullary nail (RTN) combined with less invasive stabilization system (LISS) in the treatment of distal femoral fractures.Methods:A retrospective analysis was conducted on the data of 11 patients with distal femoral fractures who underwent RTN combined with LISS locking plate treatment at the 909th Hospital from June 2021 to December 2022. There were 5 males and 6 females, with an average age of 56.5±17.5 years. The fracture types were AO/OTA 33A2 in 2 cases, A3 in 2 cases, C1 in 2 cases, and C2 in 2 cases. There were 3 cases of periprosthetic femoral fractures after total knee arthroplasty (TKA), all classified as Rorabeck type Ⅱ. They were treated with retrograde intramedullary nailing of the tibia combined with a minimally invasive lateral internal fixation system. The operation time, intraoperative blood loss, fracture healing time, full weight-bearing time, femorotibial angle, range of motion (ROM) of the knee joint, Hospital for Special Surgery (HSS) score, visual analogue scale (VAS) for pain, and complications were recorded.Results:The operation was successfully completed in all 11 cases. The operation time was 98.3±9.0 min (range 83-115 min), and the intraoperative blood loss was 167.8±24.3 ml (range 120-210 ml). All 11 cases were followed up for 11.0±1.9 months (range 9-15 months). The healing time of 11 cases was 5.3±0.8 months (range 4-6 months) after operation. The time of complete weight-bearing activity after operation was 55.7±6.5 d (range 46-67 d). At the last follow-up, the femoral-tibial angle of 11 cases was 171.2°±1.8° (range 169°-174°), the ROM of knee joint was 129.5°±4.7° (range 120°-135°), and the HSS score was 86.8±6.9 points (range 69-95 points). There were no major complications except for one case of superficial infection of surgical incision after operation.Conclusion:RTN combined with lateral LISS locking plate showed good clinical effect with the advantages of less trauma, reliable fixation, rapid recovery, less postoperative complications.

Objective:To evaluate the clinical efficacy of a novel surgical approach of debridement, antibiotics, and implant retention (DAIR) with flap transfer, for treating chronic implant infections in bone tumor patients.Methods:A retrospective review was conducted on nine consecutive patients [6 males, 3 females; median age 35(27, 51) years, range 9-71] who underwent a modified procedure of DAIR plus flap transfer between November 2022 and January 2024. The cohort included six cases of chronic periprosthetic joint infection and three cases of chronic plate and screw infection. Tumor diagnoses included seven primary malignant tumors (osteosarcoma=5, undifferentiated pleomorphic sarcoma of bone=1, synovial sarcoma=1) and two bone metastasis of renal cell carcinoma. The procedure involved wide, radical debridement, meticulous removal of biofilm from implants and surrounding soft tissue, followed by the transfer of a well vascularized musculocutaneous flap to fully envelope the contaminated interface. Pre-operative clinicopathological data, surgical details, postoperative complications and infection recurrence were analyzed.Results:The median interval between initial implantation and debridement was 10.0(3.3, 14.8) months. Median follow-up after debridement was 15.9(15.4, 18.2) months. All nine surgeries were completed as planned: six musculocutaneous flaps, two fasciocutaneous flaps and one muscle-only flap. Implants were preserved in six patients; two required subsequent removal for recurrent infection, and one patient later underwent amputation for tumor recurrence. Infection-free implant survival at 3, 6 and 12 months was 88.9%, 87.5% and 87.5%, respectively. Major complications included one donor-site hematoma, one donor-site sensory deficit and one wound healing delay. All the complications were well management. Both reinfections occurred in proximal tibial prostheses, likely due to limited flap coverage options and local anatomical constraints.Conclusion:Although reinfections happened in two cases DAIR with flap transfer provides promising short-term infection control in patients with chronic implant-associated infections following bone tumor surgery.

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 establish a comprehensive evaluation index system based on government supervision for private hospitals to standardize their management and guide their healthy operation with standardized, quantifiable assessment criteria.Methods:Through literature review, regulatory challenges and key issues in private hospital management were identified. Drawing on public hospital performance evaluations and hospital accreditation standards, a three-tiered evaluation index system was developed across four dimensions: legal establishment and practice, service capability and quality & safety, operational management, and comprehensive evaluation. The Delphi method was employed to screen indicators and determine their weights, followed by empirical research to validate the system.Results:The response rate for both rounds of expert consultation was 100%, with an authority coefficient>0.85. Indicators were optimized based on the following criteria: mean scores for importance, feasibility, and sensitivity>3.5; coefficient of variation<0.1; and Kendall′s coefficient of concordance P<0.05. The final comprehensive evaluation system consisted of 4 first-level indicators, 10 second-level indicators, and 49 third-level indicators. The evaluation of 6 private hospitals was basically consistent with the actual situation of the hospitals and the understanding of the health management department on the hospitals. Conclusions:The validated evaluation index system could provide a multi-dimensional, objective assessment of private hospitals, offering quantifiable criteria for management and serving as a reference for government oversight to promote standardized development.