Objective:To evaluate the relationship between superoxide dismutase 2 (SOD2) lactylation and nuclear receptor coactivator 4 (NCOA4)-mediated ferritinophagy-ferroptosis during cerebral ischemia-reperfusion (IR) in mice.Methods:Thirty-six clean-grade male C57BL/6 mice, aged 8-10 weeks, weighing 22-25 g, were divided into 4 groups ( n=9 each) using a table of random numbers: sham operation group (Sham group), cerebral IR group (IR group), IR+ glycolysis inhibitor 2-DG group (IR+ 2-DG group), and IR+ 2-DG+ NCOA4 overexpression group (IR+ 2-DG+ LvNCOA4 group). The model of cerebral IR injury was established by occlusion of the middle cerebral artery for 1 h followed by 24 h of reperfusion using the intraluminal suture method in anesthetized animals. 2-DG 250 mg/kg was intraperitoneally injected at 90 min before ischemia in IR+ 2-DG and IR+ 2-DG+ LvNCOA4 groups. The lentivirus overexpressing NCOA4 2 μl was injected into the ventricles at 7 days before ischemia in IR+ 2-DG+ LvNCOA4 group. The percentage of cerebral infarct volume was determined, the viable neurons were counted, and the levels of reactive oxygen species (ROS), malondialdehyde (MDA), and glutathione (GSH) were measured by enzyme-linked immunosorbent assay. The expression of SOD2, lysine 114 lactylation of superoxide dismutase 2 (SOD2-K114la), NCOA4, microtubule-associated protein 1 light chain 3β (LC3B), and acyl-CoA synthetase long-chain family member 4 (ACSL4) was determined by Western blot. Mitochondrial morphology was examined by electron microscopy. Results:Compared with Sham group, the percentage of cerebral infarct volume was significantly increased, the number of viable neurons was decreased, the levels of ROS and MDA were elevated, the content of GSH was reduced, the expression of SOD2-K114la, NCOA4, LC3B and ACSL4 was up-regulated, the expression of SOD2 was down-regulated ( P<0.05), and the mitochondrial injury was aggravated in IR group. Compared with IR group, the percentage of cerebral infarct volume was significantly decreased, the number of viable neurons was increased, the mitochondrial injury was alleviated, the levels of ROS and MDA were decreased, the content of GSH was increased, the expression of SOD2-K114la, NCOA4 and ACSL4 was down-regulated, and the expression of SOD2 and LC3B was up-regulated in IR+ 2-DG group ( P<0.05). Compared with IR+ 2-DG group, the percentage of cerebral infarct volume was significantly increased, the number of viable neurons was decreased, the levels of ROS and MDA were elevated, the content of GSH was reduced, and the expression of NCOA4, LC3B and ACSL4 was up-regulated ( P<0.05), no significant change was found in the expression of SOD2 and SOD2-K114la ( P>0.05), and the mitochondrial injury was aggravated in IR+ 2-DG+ LvNCOA4 group. Conclusions:SOD2 lactylation promotes NCOA4-mediated ferritinophagy-ferroptosis by enhancing oxidative stress, thereby contributing to the cerebral IR injury in mice.

Objective To explore the clinical value of high-frequency ultrasound combined with minimally invasive plate osteosynthesis(MIPO)in preserving the supraclavicular nerve in clavicle fractures.Methods Sixty patients with mid-clavicle fractures who were treated at the First Affiliated Hospital of Ningbo University from September 2022 to May 2024 were selected.Patients were divided into minimally invasive incision group(n=30)treated with high-frequency ultrasound combined with MIPO and traditional incision group(n=30)treated with open reduction and locked plate fixation.The perioperative indicators,visual analogue scale scores for pain,Constant-Murley shoulder scoring scale(CMS),and incidence of complications were compared between two groups.Results All patients completed the surgery and were followed up for 6 months.The bone call formation time and incision length in minimally invasive incision group were lower than those in traditional incision group(P＜0.05);The CMS at 3 months and 6 months after surgery in minimally invasive incision group were higher than those in traditional incision group(P＜0.05).The incidence of skin sensory abnormalities in minimally invasive incision group at 2 weeks,1 month,and 3 months after surgery was lower than that in traditional incision group(P＜0.05).Conclusion Compared with traditional open reduction and plate internal fixation,the high-frequency ultrasound combined with MIPO technique can effectively reduce the iatrogenic injury of the supraclavicular nerve during surgery,reduce the incidence of postoperative skin sensory abnormalities,and also have significant advantages in terms of shoulder aesthetics,shoulder joint function,and fracture healing.

Objective:To evaluate the relationship between superoxide dismutase 2 (SOD2) lactylation and nuclear receptor coactivator 4 (NCOA4)-mediated ferritinophagy-ferroptosis during cerebral ischemia-reperfusion (IR) in mice.Methods:Thirty-six clean-grade male C57BL/6 mice, aged 8-10 weeks, weighing 22-25 g, were divided into 4 groups ( n=9 each) using a table of random numbers: sham operation group (Sham group), cerebral IR group (IR group), IR+ glycolysis inhibitor 2-DG group (IR+ 2-DG group), and IR+ 2-DG+ NCOA4 overexpression group (IR+ 2-DG+ LvNCOA4 group). The model of cerebral IR injury was established by occlusion of the middle cerebral artery for 1 h followed by 24 h of reperfusion using the intraluminal suture method in anesthetized animals. 2-DG 250 mg/kg was intraperitoneally injected at 90 min before ischemia in IR+ 2-DG and IR+ 2-DG+ LvNCOA4 groups. The lentivirus overexpressing NCOA4 2 μl was injected into the ventricles at 7 days before ischemia in IR+ 2-DG+ LvNCOA4 group. The percentage of cerebral infarct volume was determined, the viable neurons were counted, and the levels of reactive oxygen species (ROS), malondialdehyde (MDA), and glutathione (GSH) were measured by enzyme-linked immunosorbent assay. The expression of SOD2, lysine 114 lactylation of superoxide dismutase 2 (SOD2-K114la), NCOA4, microtubule-associated protein 1 light chain 3β (LC3B), and acyl-CoA synthetase long-chain family member 4 (ACSL4) was determined by Western blot. Mitochondrial morphology was examined by electron microscopy. Results:Compared with Sham group, the percentage of cerebral infarct volume was significantly increased, the number of viable neurons was decreased, the levels of ROS and MDA were elevated, the content of GSH was reduced, the expression of SOD2-K114la, NCOA4, LC3B and ACSL4 was up-regulated, the expression of SOD2 was down-regulated ( P<0.05), and the mitochondrial injury was aggravated in IR group. Compared with IR group, the percentage of cerebral infarct volume was significantly decreased, the number of viable neurons was increased, the mitochondrial injury was alleviated, the levels of ROS and MDA were decreased, the content of GSH was increased, the expression of SOD2-K114la, NCOA4 and ACSL4 was down-regulated, and the expression of SOD2 and LC3B was up-regulated in IR+ 2-DG group ( P<0.05). Compared with IR+ 2-DG group, the percentage of cerebral infarct volume was significantly increased, the number of viable neurons was decreased, the levels of ROS and MDA were elevated, the content of GSH was reduced, and the expression of NCOA4, LC3B and ACSL4 was up-regulated ( P<0.05), no significant change was found in the expression of SOD2 and SOD2-K114la ( P>0.05), and the mitochondrial injury was aggravated in IR+ 2-DG+ LvNCOA4 group. Conclusions:SOD2 lactylation promotes NCOA4-mediated ferritinophagy-ferroptosis by enhancing oxidative stress, thereby contributing to the cerebral IR injury in mice.

Objective To explore the clinical value of high-frequency ultrasound combined with minimally invasive plate osteosynthesis(MIPO)in preserving the supraclavicular nerve in clavicle fractures.Methods Sixty patients with mid-clavicle fractures who were treated at the First Affiliated Hospital of Ningbo University from September 2022 to May 2024 were selected.Patients were divided into minimally invasive incision group(n=30)treated with high-frequency ultrasound combined with MIPO and traditional incision group(n=30)treated with open reduction and locked plate fixation.The perioperative indicators,visual analogue scale scores for pain,Constant-Murley shoulder scoring scale(CMS),and incidence of complications were compared between two groups.Results All patients completed the surgery and were followed up for 6 months.The bone call formation time and incision length in minimally invasive incision group were lower than those in traditional incision group(P＜0.05);The CMS at 3 months and 6 months after surgery in minimally invasive incision group were higher than those in traditional incision group(P＜0.05).The incidence of skin sensory abnormalities in minimally invasive incision group at 2 weeks,1 month,and 3 months after surgery was lower than that in traditional incision group(P＜0.05).Conclusion Compared with traditional open reduction and plate internal fixation,the high-frequency ultrasound combined with MIPO technique can effectively reduce the iatrogenic injury of the supraclavicular nerve during surgery,reduce the incidence of postoperative skin sensory abnormalities,and also have significant advantages in terms of shoulder aesthetics,shoulder joint function,and fracture healing.

Objectives:This study aimed to assess the efficacy and safety of bendamustine in combination with rituximab (BR regimen) for the treatment of newly diagnosed indolent B-cell non-Hodgkin's lymphoma (B-iNHL) and elderly mantle cell lymphoma (eMCL) .Methods:From December 1, 2020 to September 10, 2022, a multi-center prospective study was conducted across ten Grade A tertiary hospitals in Shandong Province, China. The BR regimen was administered to evaluate its efficacy and safety in newly diagnosed B-iNHL and eMCL patients, and all completed at least four cycles of induction therapy.Results:The 72 enrolled patients with B-iNHL or MCL were aged 24-74 years, with a median age of 55 years. Eastern Cooperative Oncology Group (ECOG) performance status scores of 0-1 were observed in 76.4% of patients, while 23.6% had scores of 2. Disease distribution included follicular lymphoma (FL) (51.4% ), marginal zone lymphoma (MZL) (33.3% ), eMCL (11.1% ), and the unknown subtype (4.2% ). According to the Ann Arbor staging system, 16.7% and 65.3% of patients were diagnosed with stage Ⅲ and stage Ⅳ lymphomas, respectively. Following four cycles of BR induction therapy, the overall response rate was 98.6%, with a complete response (CR) rate of 83.3% and a partial response (PR) rate of 15.3%. Only one eMCL patient experienced disease progression during treatment, and only one FL patient experienced a relapse. Even when evaluated using CT alone, the CR rate was 63.9%, considering the differences between PET/CT and CT assessments. The median follow-up duration was 11 months (range: 4-22), with a PFS rate of 96.8% and an OS rate of 100.0%. The main hematologic adverse reactions included grade 3-4 leukopenia (27.8%, with febrile neutropenia observed in 8.3% of patients), grade 3-4 lymphopenia (23.6% ), grade 3-4 anemia (5.6% ), and grade 3-4 thrombocytopenia (4.2% ). The main non-hematologic adverse reactions such as fatigue, nausea/vomiting, rash, and infections occurred in less than 20.0% of patients.Conclusion:Within the scope of this clinical trial conducted in China, the BR regimen demonstrated efficacy and safety in treating newly diagnosed B-iNHL and eMCL patients.

Objective:To evaluate the relationship between the second messenger cyclic GMP-AMP (cGAS)-cyclic GMP-AMP receptor stimulator of interferon genes (STING) signaling pathway and ferritinophagy in the early stage of cerebral ischemia-reperfusion (I/R) in mice.Methods:Twenty-four clean-grade healthy male C57BL/6 mice, aged 6-8 weeks, weighing 21-25 g, were divided into 4 groups ( n=6 each) using a random number table method: sham group, cerebral I/R injury group (CIRI group), cerebral I/R injury + cGAS inhibitor group (CIRI + RU group), and cerebral I/R injury + cGAS inhibitor + overexpressed nuclear receptor coactivator 4 (NCOA4) group (MCAO + RU + LV-NCOA4 group). The model of cerebral I/R injury was developed using the middle cerebral artery occlusion (MCAO) in anesthetized animals.In CIRI+ RU group, cGAS inhibitor 5 mg/kg was intraperitoneally injected at 10 min before reperfusion.In CIRI+ RU+ LV-NCOA4 group, NCOA4-overexpressing lentivirus (1×10 9 TU/ml) 2 μl was injected into the ventricle at 7 days before MCAO, and the other operations were the same as those previously described in CIRI+ RU group.After 6 h of reperfusion, the neurological function deficits were assessed and scored, then the mice were sacrificed, and brains were removed for determination of the cerebral infarct size (by TTC method), MDA content (by TBA method), activity of SOD (by WST-1 method), and expression of cGAS, STING, NCOA4, ferritin, and microtubule-associated protein 1 light chain 3B (LC3B) (by Western blot). Results:Compared with Sham group, the neurological function deficit score and cerebral infarct size were significantly increased, SOD activity was decreased, MDA content was increased, the expression of cGAS, STING, NCOA4 and LC3B was up-regulated, and the expression of ferritin was down-regulated in CIRI group ( P<0.05). Compared with CIRI group, the neurological function deficit score and cerebral infarct size were significantly decreased, SOD activity was increased, MDA content was decreased, the expression of cGAS, STING, NCOA4 and LC3B was down-regulated, and the expression of ferritin was up-regulated in CIRI+ RU group ( P<0.05). Compared with CIRI+ RU group, the neurological function deficit score and cerebral infarct size were significantly increased, SOD activity was decreased, MDA content was increased, the expression of cGAS, STING, NCOA4 and LC3B was up-regulated, and the expression of ferritin was down-regulated in CIRI group ( P<0.05), and no significant change was found in the expression of cGAS and STING in CIRI+ RU+ LV-NCOA4 group ( P>0.05). Conclusions:The cGAS-STING signaling pathway can promote the over-activation of ferritinophagy, enhance oxidative stress, and thus induce early CIRI in mice.

Cabozantinib, mainly targeting cMet and vascular endothelial growth factor receptor 2, is the second-line treatment for patients with advanced hepatocellular carcinoma (HCC). However, the lower response rate and resistance limit its enduring clinical benefit. In this study, we found that cMet-low HCC cells showed primary resistance to cMet inhibitors, and the combination of cabozantinib and mammalian target of rapamycin (mTOR) inhibitor, rapamycin, exhibited a synergistic inhibitory effect on the in vitro cell proliferation and in vivo tumor growth of these cells. Mechanically, the combination of rapamycin with cabozantinib resulted in the remarkable inhibition of AKT, extracellular signal-regulated protein kinases, mTOR, and common downstream signal molecules of receptor tyrosine kinases; decreased cyclin D1 expression; and induced cell cycle arrest. Meanwhile, rapamycin enhanced the inhibitory effects of cabozantinib on the migration and tubule formation of human umbilical vascular endothelial cells and human growth factor-induced invasion of cMet inhibitor-resistant HCC cells under hypoxia condition. These effects were further validated in xenograft models. In conclusion, our findings uncover a potential combination therapy of cabozantinib and rapamycin to combat cabozantinib-resistant HCC.
Anilides/pharmacology* ; Animals ; Carcinoma, Hepatocellular/drug therapy* ; Cell Line, Tumor ; Cell Proliferation ; Endothelial Cells/metabolism* ; Humans ; Liver Neoplasms/drug therapy* ; Pyridines/pharmacology* ; Sirolimus/pharmacology* ; Xenograft Model Antitumor Assays

Objective:To evaluate the role of B/glycogen synthase kinase-3β (Akt/GSK-3β) signaling pathway in interleukin-4 (IL-4)-induced reduction of cerebral ischemia-reperfusion (I/R) injury in mice and the relationship with autophagy.Methods:Forty clean-grade healthy male Balb/c mice, aged 10-12 weeks, weighing 20-25 g, were divided into 4 groups ( n=10 each) using a random number table method: sham operation group (group S), cerebral I/R group (group IR), IR plus IL-4 group, and IR plus IL-4 plus Akt inhibitor LY294002 group (IR+ IL-4+ LY group). Cerebral I/R was induced by 60 min middle cerebral artery occlusion followed by 24 reperfusion in anesthetized mice.IL-4 compound solution 0.2 ml was intraperitoneally given at 30 min before establishing the model in group IL-4.IL-4 compound solution 0.2 ml was intraperitoneally given at 30 min before establishing the model, and LY294002 15 nmol/kg was simultaneously injected via the tail vein in group IR+ IL-4+ LY.Neurological function was assessed and scored at 24 h of reperfusion, and then animals were sacrificed and brains removed for determination of cerebral infarct size (by TTC assay), cell apoptosis, autophagosome count (with transmission electron microscope), levels of superoxide dismutase (SOD), malondialdehyde (MDA) and reactive oxygen species (ROS) (using colorimetric assay), phosphorylation of Akt and GSK-3β, expression of LC3 and Beclin-1 (by Western blot). The apoptosis index and LC3Ⅱ/LC3Ⅰ ratio were calculated. Results:Compared with Sham group, the neurological scores, cerebral infarct size and apoptosis index were significantly increased, the SOD activity in brain tissues was decreased, levels of MDA and ROS were increased, phosphorylation of Akt and GSK-3β was decreased, and LC3Ⅱ/LC3Ⅰ ratio, Beclin-1 expression and autophagosome count were increased in the other three groups ( P<0.05). Compared with IR group, the neurological scores, cerebral infarct size and apoptosis index were significantly decreased, the SOD activity in brain tissues was increased, levels of MDA and ROS were decreased, phosphorylation of Akt and GSK-3β was increased, and LC3Ⅱ/LC3Ⅰ ratio, Beclin-1 expression and autophagosome count were decreased in IR+ IL-4 group ( P<0.05), and no significant change was found in the parameters mentioned above in IR+ IL-4+ LY group ( P>0.05). Compared with IR+ IL-4 group, the neurological scores, cerebral infarct size and apoptosis index were significantly increased, the SOD activity in brain tissues was decreased, levels of MDA and ROS were increased, phosphorylation of Akt and GSK-3β was decreased, and LC3Ⅱ/LC3Ⅰ ratio, Beclin-1 expression and autophagosome count were increased in IR+ IL-4+ LY group ( P<0.05). Conclusion:IL-4 can inhibit cell autophagy through activating Akt/GSK-3β signaling pathway and thus reduces cerebral I/R injury in mice.

Objective:To study the orthopedic treatment strategy for hemophilia complicated with musculoskeletal disorders as well as the peri-operative consumption of clotting factor.Methods:Total 338 orthopedic surgeries were performed for 261 patients, average age of 30.6 y (6-65 y) , with hemophilia between January 1996 and December 2019 at our institute. Two hundred and twenty-six patients presented with bleeds within the joints. Sixty-one patients presented with intramuscular bleeds, 45 presented with hemophilic pseudotumors, and six presented with miscellaneous complaints. Strategy of clotting factor replacement therapy was designed as per differences in the level of the operation procedure. Information regarding clinical manifestation, operative strategy, clotting factor consumption, and re-operation for complications was retrospectively recorded. The costs for multiple joint procedure and single joint procedure were studied.Results:We found that 270 of the 338 surgical procedures were major surgical procedures (79.9%) . There were 203 procedures of joint arthroplasty (60%) . Fourteen patients underwent reoperations for local recurrence (4.2%) . The average factor Ⅷ consumption before the surgery was 44.4 ± 8.1 IU/kg. The average FⅧ consumption within postoperative 2 weeks was 40 962 IU (647±177 IU/kg) . Seven type A hemophilic patients developed F Ⅷ inhibitor following the surgical procedure, with an average level of 13.7±11.2 BU/mL. Sixty-eight patients underwent multiple joint procedures under one anesthesia session (26%) . There was no significant difference in the factor consumption between the multiple joint procedure and single joint procedure.Conclusions:Surgical treatment was found to be effective for hemophilic arthropathy and lesion of the musculoskeletal apparatus, with the clotting factor replacement therapy. Multiple joint procedures under one anesthesia were more cost effective for patients with hemophilia, with less factor consumption than staged single joint procedure.

OBJECTIVE： To investigate and analyze the current status and existing problems of Chinese patent medicine instructions in outpatient department of our hospital， and to provide suggestions for the improvement of Chinese patent medicine instructions. METHODS： A total of 435 copies of Chinese patent medicines instructions using in the outpatient pharmacy of our hospital were collected in 2018. The labeling of usage and dosage of the instructions and other items were not clear and missing items were analyzed statistically. RESULTS： In 435 copies of drug instructions， unclear usage and dosage included usage and dosage were marked only in grams or milliliters （54 kinds， 12.4％）； daily dosage was not clear （165 kinds， 37.9％）； the words “or follow the doctor’s advice” were involved in drug instructions （86 kinds， 19.8％）； the labeling of usage and dosage for special population were not clear （34 kinds， 7.8％）； medication time was not labeled （365 kinds， 83.9％）. Unclear labeling of other items included unclear drug interaction （121 kinds， 27.8％）， unclear matters needing attention （12 kinds， 2.8％）， unclear ADR （307 kinds， 70.6％）， unclear contraindications （257 kinds， 59.1％） and unclear indications （1 kind， 0.2％）. The missing items included that drug dosage for special population （41 kinds， 94.5％）， pharmacological and toxicological items （305 kinds， 70.1％）， clinical trial data （395 kinds， 90.8％）， storage temperature label （377 kinds， 86.7％）. CONCLUSIONS： Missing items and unclear information on safe medication are common in Chinese patent medicine instructions， which need to be standardized and perfected in order to provide reference for rational drug use and guarantee the safety of drug use in patients.