ObjectiveTo investigate the effect of neuromuscular electrical stimulation (NMES) on quadriceps muscle strength and walking for patients after anterior cruciate ligament reconstruction (ACLR). MethodsThirty-four patients after ACLR were selected at Beijing Rehabilitation Hospital of Capital Medical University from July, 2022 to October, 2023, and randomly divided into control group (n = 17) and experimental group (n = 17). Both groups received routine rehabilitation and functional training, and the experimental group received NMES during the functional training, while the control group received sham NMES, for eight weeks. Quadriceps peak torque-to-weight ratio, single-leg support phase and plantar impulses during walking were measured before and after intervention. ResultsTwo cases in the control group and three in the experimental group dropped down. Quadriceps peak torque-to-weight ratio improved in both groups after intervention (|t| > 17.578, P < 0.001), and improved more in the experimental group than in the control group (t = 4.714, P < 0.001); while the affected single-leg support phase and the affected/unaffected single-leg support phase ratio improved in both groups (|t| > 16.882, P < 0.001), and improved more in the experimental group than in the control group (t > 3.234, P < 0.01); and plantar impulses of all zones optimized in both groups (t > 9.221, P < 0.001), and were better in the experimental group than in the control group(|t| > 2.852, P < 0.01). ConclusionNMES may further improve quadriceps muscle strength, plantar pressure distribution during walking and single-leg support in patients after ACLR.

Objective:Chimeric antigen receptor T-cell(CAR-T)immunotherapy has made major breakthroughs in the treatment of blood tu-mors.However,current CAR-T therapies face several limitations:they require autologous cells,involve a lengthy and costly production pro-cess,and use lentiviral transduction that carry risk of insertional carcinogenesis due to random integration.Therefore,there is an urgent need to develop a universal cost-effective cancer immunotherapy method generating CAR-T cells for in vivo cancer immunotherapy.Meth-ods:This study successfully established an exosome-mediated,T-cell targeted delivery system,demonstrating both precise design and func-tional efficacy for biomedical applications.To optimize CAR-T cell generation the transfection dose was adjusted,and the kinetics of CAR-T cell percentage were recorded.The cytotoxicity of the resulting CAR-T cells was evaluated in vitro by calcein-AM release.To test the tumor-killing in vivo of engineered exosomes,human PBMCs were injected into NPG mice via the tail vein to establish humanized mice,followed by intravenous injection of tumor cells to induce cancer.Results:To overcome the limitations of conditional autologous CAR-T cells,we de-veloped a T cell-targeted exosome system capable of specifically targeting human CD3+,CD4+,and CD8+T cells.CAR-T production was dose-dependent,with transfection efficiency reaching upto 97.8%at 106 particles/cell.Both in vitro cytotoxicity assays and in vivo animal experi-ments demonstrated that exosome-incubated CAR-T cells effectively eliminated CD19-positive Raji cells,highlighting their specificity and therapeutic potential in antigen-directed applications.Conclusions:We successfully established a CD8-targeting exosome delivery system for CAR-T cell production capable of transforming CD8+T cells into functional CAR-T cells,which showed significant tumor-killing ability in vitro and in mice.Compared with the traditional lentiviral vector for the preparation of CAR-T cells in vitro,in vivo-reprogrammed CAR-T cells us-ing our CD8-targeted exosome delivery system,with higher transfection efficiency,shorter production period,lower cost,and eliminated the risk of insertion carcinogenesis.This strategy promises to bring a new era of universal CAR-T medicine,which can improve cancer immuno-therapy and may hold promise as a therapeutic platform to treat various diseases.

Objective:Chimeric antigen receptor T-cell(CAR-T)immunotherapy has made major breakthroughs in the treatment of blood tu-mors.However,current CAR-T therapies face several limitations:they require autologous cells,involve a lengthy and costly production pro-cess,and use lentiviral transduction that carry risk of insertional carcinogenesis due to random integration.Therefore,there is an urgent need to develop a universal cost-effective cancer immunotherapy method generating CAR-T cells for in vivo cancer immunotherapy.Meth-ods:This study successfully established an exosome-mediated,T-cell targeted delivery system,demonstrating both precise design and func-tional efficacy for biomedical applications.To optimize CAR-T cell generation the transfection dose was adjusted,and the kinetics of CAR-T cell percentage were recorded.The cytotoxicity of the resulting CAR-T cells was evaluated in vitro by calcein-AM release.To test the tumor-killing in vivo of engineered exosomes,human PBMCs were injected into NPG mice via the tail vein to establish humanized mice,followed by intravenous injection of tumor cells to induce cancer.Results:To overcome the limitations of conditional autologous CAR-T cells,we de-veloped a T cell-targeted exosome system capable of specifically targeting human CD3+,CD4+,and CD8+T cells.CAR-T production was dose-dependent,with transfection efficiency reaching upto 97.8%at 106 particles/cell.Both in vitro cytotoxicity assays and in vivo animal experi-ments demonstrated that exosome-incubated CAR-T cells effectively eliminated CD19-positive Raji cells,highlighting their specificity and therapeutic potential in antigen-directed applications.Conclusions:We successfully established a CD8-targeting exosome delivery system for CAR-T cell production capable of transforming CD8+T cells into functional CAR-T cells,which showed significant tumor-killing ability in vitro and in mice.Compared with the traditional lentiviral vector for the preparation of CAR-T cells in vitro,in vivo-reprogrammed CAR-T cells us-ing our CD8-targeted exosome delivery system,with higher transfection efficiency,shorter production period,lower cost,and eliminated the risk of insertion carcinogenesis.This strategy promises to bring a new era of universal CAR-T medicine,which can improve cancer immuno-therapy and may hold promise as a therapeutic platform to treat various diseases.

Objective:To investigate the efficacy and safety of kidney sparing treatment based on Thulium laser ablation and systematic therapy in localized high-risk upper urinary tract urothelial carcinoma (UTUC).Methods:The data of 10 patients with UTUC who received combined treatment based on Thulium laser and systematic treatment from January 2020 to December 2021 in West China Hospital were retrospectively analysed. There were 5 males and 5 females with a median age of 76 (range 52 to 87)years old. Three cases were renal pelvis tumor and 7 cases were ureter tumor including 5 cases in lower ureter and 2 cases in upper and middle ureter. Five cases were with positive urine cytology and 6 cases were with hydronephrosis. One case was muscular invasion UTUC confirmed by biopsy(cT 2+), 7 cases were high-grade invasive urothelial carcinoma (cT 1+), and 2 cases were high-grade papillary urothelial carcinoma (cT a). Among 10 cases, 5 patients refused radical nephroureterectomy(RUN), among whom 3 patients were too old or in poor general condition to tolerate RNU. One case had a solitary kidney and 1 case had bilateral tumours. Patients were treated with Thulium laser tumor ablation under ureteroscopy combined with systemic therapy. The perioperative systemic treatment included platinum-based chemotherapy±immunotherapy, RC48+ immunotherapy, and immunotherapy alone. The postoperative treatment was immunotherapy maintenance±local radiotherapy. Strict follow-up was conducted after the completion of treatment. Results:Nine patients received systemic therapy before ablation. Four cycles of platinum-based chemotherapy (cisplatin in 2 cases, carboplatin in 1 case) were used in 3 cases, and platinum-based chemotherapy + immunotherapy (6 cycles of cisplatin + toripalimab in 1 case, 4 cycles of cisplatin + toripalimab in 1 case, 4 cycles of carboplatin+ trelizumab in 1 case) was used in 3 cases, four cycle of RC48 + immunotherapy (toripalimab or trelizumab) were used in 2 cases, and four cycles of immunotherapy (toripalimab) were used in 1 case. The operations of 10 cases were successfully completed without serious complications during the perioperative period and the laser working time (42.4 ± 15.2) min. Of the 10 cases, 4 achieved complete ablation at the first ablation, and 6 patients had incomplete ablation. Among them, 2 patients achieved clinical complete remission after 1-2 cycles of systemic therapy, and 4 patients achieved complete ablation after Thulium laser ablation again.All the 10 patients were treated with immunotherapy for 1 year, and 2 of them received additional adjuvant radiotherapy. The patients were followed-up for median 40 months(range 26 to 53 months). Recurrence occurred in 5 cases, of which 3 cases underwent salvage nephroureterectomy and 2 cases underwent Thulium laser ablation under ureteroscopy again. Five patients had no tumor recurrence. None of the 10 patients had distant metastasis. At the last follow-up, 1 patient died of complications and 6 patients kept the affected kidney alive. Perioperative complications including macroscopic hematuria (8 cases), fever (3 cases), the long-term complications of ureter stenosis (4 cases).Conclusions:For localized high-risk UTUC, local Thulium laser ablation combined with systemic therapy can achieve good tumor control while preserving the affected kidney in selected patients, and its potential application value should be further evaluated.

Objective:To document any effect of combining kinesiotaping (KT) with Monitored Rehab Systems biofeedback training (MRS training) on the recovery of knee function after anterior cruciate ligament reconstruction (ACLR).Methods:Twenty-two patients after ACLR were randomly divided into an MRS group and a combined group, each of 11. In addition to conventional rehabilitation training, the MRS group had placebo taping in place during their MRS training, while the combined group had KT applied. Each training session lasted 45 minutes, with three sessions per week. Before the experiment and after 8 weeks of treatment, the affected foot support phase, impulse of each area of the affected foot′s sole, and lower limb reaction time were measured.Results:Significant improvement in the affected foot′s support phase and lower limb reaction time was documented in both groups, as well as in the average impulse percentage of the affected foot′s plantar forefoot area (F area), mid-foot area (MF area), and heel area (H area). All were significantly better in the combined group than in the MRS group, on average.Conclusion:Supplementing MRS biofeedback training with kinesiotaping can more effectively improve the knee joint functioning and postural control of patients after ACLR surgery. The combination is worthy of clinical promotion and application.

ObjectiveTo explore the effects of different emulsion mixtures and emulsification methods on the inflammation severity in an experimental autoimmune uveitis （EAU） model in rats， and to analyze the characteristics of the current EAU model. MethodEAU was induced in Lewis rats by subcutaneous injection of interphotoreceptor retinoid-binding protein （IRBP） 1177-1191 emulsified with Freund's complete adjuvant （CFA）， with or without intraperitoneal injection of pertussis toxin （PTX）. Slit lamp examination， HE staining， and optical coherence tomography were used to evaluate factors affecting EAU modeling， including different doses of the emulsion mixture （IRBP1177-1191， PTX， and inactivated Mycobacterium tuberculosis） and four different emulsification methods. The classification， characteristics， modeling methods， advantages， and disadvantages of EAU animal models were summarized and analyzed based on the clinical diagnostic criteria and syndrome characteristics of chronic uveitis in both traditional Chinese medicine （TCM） and western medicine， to evaluate the consistency between TCM and western medical syndromes. ResultIncreasing the dose of inactivated M. tuberculosis and antigen peptide in the emulsion mixture exacerbated the anterior segment inflammation in EAU rats. Increasing the injection of PTX also exacerbated anterior segment inflammation and increased retinal thickness in EAU rats. The severity of the EAU model was closely related to the emulsification method used. All four emulsification methods successfully induced EAU in rats. Comparatively， the ultrasonic cell disruptor and T10 basic disperser achieved successful emulsification in a short time. The degree of emulsification of the mixture also influenced the severity of the EAU model in rats. The existing EAU animal model shows a high degree of consistency with western medical diagnoses and the main ocular syndromes in TCM. ConclusionIRBP1177-1191， PTX， inactivated M. tuberculosis， and emulsification methods can affect the severity of the EAU model through different pathways. The existing EAU animal models can simulate the clinical characteristics of western medicine well but lack the etiology， pathogenesis， and syndrome characteristics of TCM. Therefore， it is necessary to construct an EAU animal model that combines disease and syndrome characteristics.

Objective: To analyze the prevalence and related factors of pediatric flexible flatfoot (PFF) among 7-8 year old children in Changzhou, so as to provide a feasible basis for the prevention and treatment of PFF. Methods: From December 2023 to February 2024, a total of 1 685 children aged 7-8 from 10 primary schools in Changzhou were selected by stratified cluster random sampling method, and screened for PFF by using a foot optical assessment recording device. Information including sex, body mass index (BMI), diet, exercise and shoe wearing habits were collected. The valgus angle of the hindfoot was measured on the body surface by using an orthopedic measuring ruler in the standing position. Pain levels were evaluated by using visual analogue score (VAS) for children with flatfoot syndrome. Multivariate Logistic analysis was used to analyze related factors of PFF. Results: The overall detection rate of PFF was 27.4%, and there was a significant difference in the detection rate of PFF between boys and girls, with 30.3% and 24.1% respectively ( χ 2=7.96, P < 0.01 ). Most cases of PFF were mild flatfoot (60.8%) and bilateral ( 60.4% ). Approximately 13.2% of children with PFF had flatfoot syndrome, with a mean VAS of (2.86±0.73). About 56.1% of children with PFF had a normal valgus angle of the hindfoot. Sex, high BMI and preference for shoe last with front upturned shoe shape were positively correlated with the detection of PFF ( OR= 1.74, 1.54, 1.13, P <0.05). After stratified by sex, regular exercise in boys and age in girls were negatively correlated with the detection of PFF ( OR=0.40, 0.64, P <0.05). Conclusions The detection rate of PFF in 7-8 year old children is high. Additionally, PFF combined with flatfoot syndrome or valgus hindfoot is relatively rare and is likely to be underestimated, which emphasizes the importance of early detection and intervention for PFF.

Objective To investigate the effect of daily soft brace wearing on plantar dynamics during walking in patients with chronic ankle instability(CAI). Methods A total of 52 patients with unilateral chronic ankle instability(CAI)in Beijing Rehabilitation Hospital from February,2021 to January,2023 were randomly divided into control group(n=26)and experimental group(n=26).Both groups underwent an eight-week exercise training program.The control group wore placebo brace dur-ing daily activities,while the experimental group wore soft ankle brace.Plantar dynamic parameters were mea-sured using a pressure plate system during walking,including peak plantar pressure and plantar impulse before and after intervention. Results Six participants dropped out in the control group and five in the experimental group,resulting in a final inclu-sion of 41 participants.After intervention,there was no significant difference in peak plantar pressure and im-pulse on the affected side in the control group among different areas(P＞0.05).In the experimental group,the peak pressure and impulse in the heel medial,heel lateral and forefoot medial areas increased(|t|＞4.192,P＜0.001),while the peak pressure and impulse in the midfoot and lateral forefoot areas decreased(t＞2.984,P＜0.05);the peak pressure and impulse in the heel medial,heel lateral and forefoot medial areas were higher in the experimental group than in the control group(|t|＞2.126,P＜0.05),and the peak pressure and impulse were low-er in the midfoot and forefoot lateral areas(t＞2.133,P＜0.05). Conclusion Wearing a soft brace during daily activities may optimize the distribution of peak plantar pressure and plan-tar impulse on the affected side in patients with CAI,which may prevent recurrence of sprains.

Background::Bladder cancer, characterized by a high potential of tumor recurrence, has high lifelong monitoring and treatment costs. To date, tumor cells with intrinsic softness have been identified to function as cancer stem cells in several cancer types. Nonetheless, the existence of soft tumor cells in bladder tumors remains elusive. Thus, our study aimed to develop a microbarrier microfluidic chip to efficiently isolate deformable tumor cells from distinct types of bladder cancer cells.Methods::The stiffness of bladder cancer cells was determined by atomic force microscopy (AFM). The modified microfluidic chip was utilized to separate soft cells, and the 3D Matrigel culture system was to maintain the softness of tumor cells. Expression patterns of integrin β8 (ITGB8), protein kinase B (AKT), and mammalian target of rapamycin (mTOR) were determined by Western blotting. Double immunostaining was conducted to examine the interaction between F-actin and tripartite motif containing 59 (TRIM59). The stem-cell-like characteristics of soft cells were explored by colony formation assay and in vivo studies upon xenografted tumor models. Results::Using our newly designed microfluidic approach, we identified a small fraction of soft tumor cells in bladder cancer cells. More importantly, the existence of soft tumor cells was confirmed in clinical human bladder cancer specimens, in which the number of soft tumor cells was associated with tumor relapse. Furthermore, we demonstrated that the biomechanical stimuli arising from 3D Matrigel activated the F-actin/ITGB8/TRIM59/AKT/mTOR/glycolysis pathways to enhance the softness and tumorigenic capacity of tumor cells. Simultaneously, we detected a remarkable up-regulation in ITGB8, TRIM59, and phospho-AKT in clinical bladder recurrent tumors compared with their non-recurrent counterparts.Conclusions::The ITGB8/TRIM59/AKT/mTOR/glycolysis axis plays a crucial role in modulating tumor softness and stemness. Meanwhile, the soft tumor cells become more sensitive to chemotherapy after stiffening, that offers new insights for hampering tumor progression and recurrence.

Background::Genetic variants of dopaminergic transcription factor-encoding genes are suggested to be Parkinson’s disease (PD) risk factors; however, no comprehensive analyses of these genes in patients with PD have been undertaken. Therefore, we aimed to genetically analyze 16 dopaminergic transcription factor genes in Chinese patients with PD.Methods::Whole-exome sequencing (WES) was performed using a Chinese cohort comprising 1917 unrelated patients with familial or sporadic early-onset PD and 1652 controls. Additionally, whole-genome sequencing (WGS) was performed using another Chinese cohort comprising 1962 unrelated patients with sporadic late-onset PD and 1279 controls.Results::We detected 308 rare and 208 rare protein-altering variants in the WES and WGS cohorts, respectively. Gene-based association analyses of rare variants suggested that MSX1 is enriched in sporadic late-onset PD. However, the significance did not pass the Bonferroni correction. Meanwhile, 72 and 1730 common variants were found in the WES and WGS cohorts, respectively. Unfortunately, single-variant logistic association analyses did not identify significant associations between common variants and PD. Conclusions::Variants of 16 typical dopaminergic transcription factors might not be major genetic risk factors for PD in Chinese patients. However, we highlight the complexity of PD and the need for extensive research elucidating its etiology.