Immunotherapy has emerged as a revolutionary approach to treat immune-related diseases. Dendritic cells (DCs) play a pivotal role in orchestrating immune responses, making them an attractive target for immunotherapeutic interventions. Modulation of gene expression in DCs using genome editing techniques, such as the CRISPR-Cas system, is important for regulating DC functions. However, the precise delivery of CRISPR-based therapies to DCs has posed a significant challenge. While lipid nanoparticles (LNPs) have been extensively studied for gene editing in tumor cells, their potential application in DCs has remained relatively unexplored. This study investigates the important role of cholesterol in regulating the efficiency of BAMEA-O16B lipid-assisted nanoparticles (BLANs) as carriers of CRISPR/Cas9 for gene editing in DCs. Remarkably, BLANs with low cholesterol density exhibit exceptional mRNA uptake, improved endosomal escape, and efficient single-guide RNA release capabilities. Administration of BLAN_mCas9/gPD-L1 results in substantial PD-L1 gene knockout in conventional dendritic cells (cDCs), accompanied by heightened cDC1 activation, T cell stimulation, and significant suppression of tumor growth. The study underscores the pivotal role of cholesterol density within LNPs, revealing potent influence on gene editing efficacy within DCs. This strategy holds immense promise for the field of cancer immunotherapy, offering a novel avenue for treating immune-related diseases.

Objective:To investigate the effect of an integrated orthopedic rehabilitation model on pain,ankle function and exercise level over 24 weeks after lateral ankle ligament repair.Method:A retrospective cohort of 81 patients after ankle ligament repair at the fourth medical center of Chinese PLA general hospital from September,2021 to September,2022 were selected.The patients were divided into an experimental cohort(n=43)and a control cohort(n=38)by whether the patient had received integrated orthopaedic rehabilitation(strictly follow clinical pathway of rehabilitation for postoperative ankle ligament repair patients)or conventional physician-guided rehabilitation.American orthopedic foot and ankle society score(AOFAS)、visual analogue scale(VAS)、foot and ankle ability measure-activities of daily living(FAAM-ADL)、foot and ankle ability measure-sport(FAAM-S)and Tegner score were assessed at 2 weeks、12 weeks、24 weeks after surgery.Result:The AOFAS、VAS score was statistically higher in experimental cohort than in control cohort at 12 weeks after surgery(P=0.043、P=0.033).The AOFAS、FAAM-ADL、Tegner score(P=0.017、P=0.021、P=0.038,re-spectively)was higher and VAS score(P=0.035)was lower in the experimental cohort than in control cohort at 24 weeks.Conclusion:Integrated orthopaedic rehabilitation can improve ankle function,exercise level and quality of life on patients after ankle ligament repair surgery compared with conventional rehabilitation.

Objective: To examine the impact of temperature on the incidence of hand-foot-mouth disease (HFMD), so as to provide a basis for research on the impact of climate on HFMD. Methods: HFMD incidence data in Huzhou City, Zhejiang Province from 2010 to 2022 were collected from the Infectious Disease Surveillance System of Chinese Disease Prevention and Control Information System. Concurrent meteorological data were obtained from the Huzhou Meteorological Bureau. A distributed lag non-linear model (DLNM) was employed to analyze the impact of temperature on HFMD incidence. Results: A total of 65 059 cases of HFMD were reported in Huzhou City from 2010 to 2022, with incidence peaks occurring from April to July and from October to November. The 5th, 25th, 50th, 75th, and 95th percentiles (P5, P25, P50, P75, and P95) of daily average temperature were 2.5, 9.4, 17.8, 24.7, and 30.6 ℃, respectively. The results from the DLNM analysis revealed a non-linear relationship between daily average temperature and HFMD incidence, which exhibited an irregular "M" shape. The incidence peaks were observed at 6.5 ℃ (RR=1.596, 95%CI: 1.154-2.207) and 25.7 ℃ (RR=3.257, 95%CI: 2.614-4.058), respectively. The maximum single-day effect was observed at a daily average temperature of -6.2 ℃ at lag 0 day (RR=1.250, 95%CI: 1.100-1.420), while the minimum single-day effect was observed at a daily average temperature of 34.6 ℃ at lag 0 day (RR=0.881, 95%CI: 0.826-0.940). The impact of daily average temperature at P5 on the incidence of HFMD showed RR values greater than 1 at lag 0, 3, 27, and 30 days, but less than 1 at lag 9 and 12 days, with no significant cumulative lag effect. In contrast, for the daily average temperature at P95, the RR values less than 1 at lag 15, 18, and 21 days, with significant cumulative lag effects identified. Conclusions The incidence of HFMD in Huzhou City peaks from April to July and again from October to November. There exists a nonlinear relationship and lag effect between temperature and HFMD incidence, with particular attention drawn to the impact of extreme temperatures on the disease's onset.

Diabetic cardiomyopathy (DCM) is a medical condition characterized by cardiac remodeling and dysfunction in individuals with diabetes mellitus. Sarcoplasmic reticulum (SR) and mitochondrial Ca²⁺ overload in cardiomyocytes have been recognized as biological hallmarks in DCM; however, the specific factors underlying these abnormalities remain largely unknown. In this study, we aimed to investigate the role of a cardiac-specific long noncoding RNA, D830005E20Rik (Trdn-as), in DCM. Our results revealed the remarkably upregulation of Trdn-as in the hearts of the DCM mice and cardiomyocytes treated with high glucose (HG). Knocking down Trdn-as in cardiac tissues significantly improved cardiac dysfunction and remodeling in the DCM mice. Conversely, Trdn-as overexpression resulted in cardiac damage resembling that observed in the DCM mice. At the cellular level, Trdn-as induced Ca²⁺ overload in the SR and mitochondria, leading to mitochondrial dysfunction. RNA-seq and bioinformatics analyses identified calsequestrin 2 (Casq2), a primary calcium-binding protein in the junctional SR, as a potential target of Trdn-as. Further investigations revealed that Trdn-as facilitated the recruitment of METTL14 to the Casq2 mRNA, thereby enhancing the m⁶A modification of Casq2. This modification increased the stability of Casq2 mRNA and subsequently led to increased protein expression. When Casq2 was knocked down, the promoting effects of Trdn-as on Ca²⁺ overload and mitochondrial damage were mitigated. These findings provide valuable insights into the pathogenesis of DCM and suggest Trdn-as as a potential therapeutic target for this condition.
Animals ; Diabetic Cardiomyopathies/pathology* ; RNA, Long Noncoding/genetics* ; Myocytes, Cardiac/metabolism* ; Mice ; Calsequestrin/genetics* ; Calcium/metabolism* ; Male ; Sarcoplasmic Reticulum/metabolism* ; Methyltransferases/metabolism* ; Mice, Inbred C57BL ; Mitochondria, Heart/metabolism* ; Disease Models, Animal ; Mitochondria/metabolism*

Objective:Utilizing the principles of mechanical vibrations dissections and low-temperature coagulation of harmonic scalpel, and combining the magnifying effect of 3D thoracoscopy and the advantage of low trauma, we used harmonic scalpel to harvest the left internal mammary artery (LIMA) under thoracoscopy, and explored the feasibility and early effect of harmonic scalpel in minimally invasive coronary artery bypass grafting (MICABG).Methods:A total of 488 patients who underwent MICABG from July 2021 to December 2023 in Dong E Hospital were collected. Among them, 347 patients was harvested using harmonic scalpel under thoracoscopy. Following anastomosis, the blood flow and pulsatility index (PI) from the graft to the target vessel were determined using a transit time flow meter (MiraQ, Medistim, Norway), and patency was checked. Data from 347 patients were analyzed.Results:All 347 patients were performed under non-extracorporeal circulation and successful graft patency was achieved in all patients. Analysis of 79 patients revealed a median time to LIMA acquisition of 32 min and a median length of acquisition of 15 cm. No graft-related events or complications occurred in any of the patients, and 2 patients died postoperatively. During a median follow-up of 12.1 months, only one patient experienced graft occlusion necessitating intervention.Conclusion:The utilization of harmonic scalpel for LIMA harvesting in MICABG is feasible and yields stable early results.

Objective:To explore the possibility of using a inferior gluteal artery perforator flap (IGAPF) for breast reconstruction in the patient who did not have suitable donor site in back and abdomen.Methods:In November 2024, a 25-year-old unmarried and childless woman with right breast cancer received immediate right breast reconstruction by a right free IGAPF after modified right mastectomy in the Department of Breast and Thyroid Surgery, Second Affiliated Hospital of Hainan Medical University. The locations of perforators were confirmed by both Multi-detector computed tomography angiography (MDCTA) and portable Doppler blood flow detector before surgery. The IGAPF was designed to take the inferior gluteal wrinkle as the lower edge, the axis of the flap was parallel to the inferior gluteal wrinkle, and the width of the flap was estimated where the incision could be directly closed. The size of right IGAPF was 6.0 cm×19.0 cm. Sharp dissection was performed between the sarcolemma and muscle fibres of gluteus, then the perforators were dissected along the direction of muscle fibres of gluteus. The vascular pedicle was kept at about 8.0 cm in length. The diameter of artery was about 2.0 mm and that for the veins was about 1.5 mm. End-to-end anastomoses with the right thoracodorsal artery and vein were successfully carried out. The donor site was directly closed, and it was hidden in the inferior gluteal wrinkle. Postoperative outpatient clinical review was made.Results:Pathological examination reported: an invasive carcinoma of right breast, axillary lymph node metastasis (2/10). The patient recovered well and the flap survived without any complication, i.e. ischemic necrosis, infection and haematoma. The patient was off-bed at 3 days and discharged at 13 days after surgery. At the 40 days of postoperative follow-up, the patient achieved a good recovery and the lower limb activity was not affected by the surgery. The patient was satisfied with the reconstructed breast and donor site recovery. The patient followed with scheduled chemotherapy and subsequent radiotherapy. The volume of reconstructed breast was smaller than the other breast, of which the patient was fully informed before the surgery.Conclusion:A free IGAPF provides an alternative donor sites for achieving a breast reconstruction due to the reliable pedicle vessels and invisible donor scars.

Porcine arthritis,one of the common chronic diseases in large-scale pig farms,can signifi-cantly reduce the production performance of meat pigs.In this study,a strain of Haemophilus pa-rasuis(HPS)was isolated from the joint fluid of a lame pig.The HPS was analyzed in terms of se-rotypes,virulence genes,and resistance genes.Additionally,it was treated with sensitive antibiotics to provide a theoretical basis for the comprehensive prevention and treatment of arthritis in meat pigs in future production settings.A strain of HPS type 14 was isolated from the joint fluid of dis-eased pigs.The HPS isolate demonstrated sensitivity to β-lactams and tetracyclines,while florfeni-col and polymyxin effectively inhibited its growth at low concentrations.However,the bacteria ex-hibited resistance to sulfonamides and ciprofloxacin.The treatment of affected pigs with clinical ar-thritis using doxycycline and enrofloxacin injections proved effective.Compared to the infected group,in which the sick pigs experienced difficulty flexing their carpal and tarsal joints and exhibi-ted significant lameness,the pigs in the treatment group showed marked improvement.Their joints were only slightly swollen,and the clinical symptoms of arthropathy were alleviated.

Objective:To investigate the effect of an integrated orthopedic rehabilitation model on pain,ankle function and exercise level over 24 weeks after lateral ankle ligament repair.Method:A retrospective cohort of 81 patients after ankle ligament repair at the fourth medical center of Chinese PLA general hospital from September,2021 to September,2022 were selected.The patients were divided into an experimental cohort(n=43)and a control cohort(n=38)by whether the patient had received integrated orthopaedic rehabilitation(strictly follow clinical pathway of rehabilitation for postoperative ankle ligament repair patients)or conventional physician-guided rehabilitation.American orthopedic foot and ankle society score(AOFAS)、visual analogue scale(VAS)、foot and ankle ability measure-activities of daily living(FAAM-ADL)、foot and ankle ability measure-sport(FAAM-S)and Tegner score were assessed at 2 weeks、12 weeks、24 weeks after surgery.Result:The AOFAS、VAS score was statistically higher in experimental cohort than in control cohort at 12 weeks after surgery(P=0.043、P=0.033).The AOFAS、FAAM-ADL、Tegner score(P=0.017、P=0.021、P=0.038,re-spectively)was higher and VAS score(P=0.035)was lower in the experimental cohort than in control cohort at 24 weeks.Conclusion:Integrated orthopaedic rehabilitation can improve ankle function,exercise level and quality of life on patients after ankle ligament repair surgery compared with conventional rehabilitation.

Porcine arthritis,one of the common chronic diseases in large-scale pig farms,can signifi-cantly reduce the production performance of meat pigs.In this study,a strain of Haemophilus pa-rasuis(HPS)was isolated from the joint fluid of a lame pig.The HPS was analyzed in terms of se-rotypes,virulence genes,and resistance genes.Additionally,it was treated with sensitive antibiotics to provide a theoretical basis for the comprehensive prevention and treatment of arthritis in meat pigs in future production settings.A strain of HPS type 14 was isolated from the joint fluid of dis-eased pigs.The HPS isolate demonstrated sensitivity to β-lactams and tetracyclines,while florfeni-col and polymyxin effectively inhibited its growth at low concentrations.However,the bacteria ex-hibited resistance to sulfonamides and ciprofloxacin.The treatment of affected pigs with clinical ar-thritis using doxycycline and enrofloxacin injections proved effective.Compared to the infected group,in which the sick pigs experienced difficulty flexing their carpal and tarsal joints and exhibi-ted significant lameness,the pigs in the treatment group showed marked improvement.Their joints were only slightly swollen,and the clinical symptoms of arthropathy were alleviated.

Objective:To explore the possibility of using a inferior gluteal artery perforator flap (IGAPF) for breast reconstruction in the patient who did not have suitable donor site in back and abdomen.Methods:In November 2024, a 25-year-old unmarried and childless woman with right breast cancer received immediate right breast reconstruction by a right free IGAPF after modified right mastectomy in the Department of Breast and Thyroid Surgery, Second Affiliated Hospital of Hainan Medical University. The locations of perforators were confirmed by both Multi-detector computed tomography angiography (MDCTA) and portable Doppler blood flow detector before surgery. The IGAPF was designed to take the inferior gluteal wrinkle as the lower edge, the axis of the flap was parallel to the inferior gluteal wrinkle, and the width of the flap was estimated where the incision could be directly closed. The size of right IGAPF was 6.0 cm×19.0 cm. Sharp dissection was performed between the sarcolemma and muscle fibres of gluteus, then the perforators were dissected along the direction of muscle fibres of gluteus. The vascular pedicle was kept at about 8.0 cm in length. The diameter of artery was about 2.0 mm and that for the veins was about 1.5 mm. End-to-end anastomoses with the right thoracodorsal artery and vein were successfully carried out. The donor site was directly closed, and it was hidden in the inferior gluteal wrinkle. Postoperative outpatient clinical review was made.Results:Pathological examination reported: an invasive carcinoma of right breast, axillary lymph node metastasis (2/10). The patient recovered well and the flap survived without any complication, i.e. ischemic necrosis, infection and haematoma. The patient was off-bed at 3 days and discharged at 13 days after surgery. At the 40 days of postoperative follow-up, the patient achieved a good recovery and the lower limb activity was not affected by the surgery. The patient was satisfied with the reconstructed breast and donor site recovery. The patient followed with scheduled chemotherapy and subsequent radiotherapy. The volume of reconstructed breast was smaller than the other breast, of which the patient was fully informed before the surgery.Conclusion:A free IGAPF provides an alternative donor sites for achieving a breast reconstruction due to the reliable pedicle vessels and invisible donor scars.