The biological activity of chitinase in degrading chitin has garnered extensive attention,particularly for its potential applications in biological control.This study utilized four spore-form-ing Bacillus strains isolated from Dermacentor nuttalli ticks collected in the Hulunbuir region.Traditional bacterial culture methods were employed for isolation and identification,followed by 16S rRNA sequencing and phylogenetic analysis of the purified cultures.chitin-hydrolyzing strains were screened using colloidal chitin plates,and specific chitinase genes were detected via PCR.Fer-mentation was conducted at 37.0 ℃ for 4 d,and the supernatants were subjected to enzyme activity analysis using the DNS method.Four Gram-positive Bacillus strains were successfully isolated from tick tissue samples,they were identified as B.proteolyticus,B.paramycoides,B.thuringien-sis,and B.cereus,and renamed IMH/B-1,IMH/P-1,IMH/T-1,and IMH/C-1,respectively.PCR a-nalysis detected chitinase genes in B.proteolyticus and B.thuringiensis,while B.cereus and B.pa-ramycoides lacked these genes.However,three strains B.proteolyticus,B.thuringiensis,and B.ce-reus demonstrated significant(P＜0.01)chitin degradation activity on colloidal chitin.Enzyme ac-tivity assays revealed that chitinase activity ranged from 1.292 to 2.032 U/mL,with B.proteolytic-us exhibiting the highest activity 2.032 U/mL,followed by B.cereus 1.496 U/mL and B.thuring-iensis 1.324 U/mL.This study provides a foundation for further research and application of chiti-nase-producing Bacillus strains.

Objective To construct a risk prediction model for venous thromboembolism(VTE)in patients with acute stroke and analyze its predictive value with decision curve.Methods Clinical data of 265 patients with acute stroke admitted to Department of Neurology of Pingxiang People's Hospital between May 2022 and May 2024 were collected,and 185 of them(69.8％)were assigned to a training set and 80 cases(30.2％)into a validation set.Based on the results of color Doppler ultrasound examination,the patients in the training set were divided into the VTE group(72 cases)and the non-VTE group(113 cases).Multivariate logistic regression analysis was used to identify the risk factors for VTE occurrence in the patients.A prediction model was constructed and a nomegram was developed to visually present the logistic regression prediction model.ROC curve and decision curve analyses were applied to evaluate the predictive performance of the logis-tic regression model.Results The VTE group had significantly larger intracranial hemorrhage volume,higher fibrinogen(FIB),D-dimer and C-reactive protein(CRP)levels,and lower Glasgow Coma Scale(GCS)score than the non-VTE group(P＜0.01).Multivariate logistic regression analysis indicated that intracranial hemorrhage volume(OR=1.243,95％CI:1.015-1.522),FIB(OR=3.161,95％CI:1.711-5.841),CRP(OR=1.595,95％CI:1.283-1.981),and D-dimer(OR=6.234,95％CI:2.693-14.434)were independent risk factors for the occurrence of VTE in the patients with acute stroke,while GCS score(OR=0.587,95％CI:0.413-0.834,P=0.003)was an independent protective factor against the occurrence.A VTE prediction model based on these influencing factors obtained a consistency index of 0.978.The calibration curve results showed that the observed values were in good agreement with the predicted values.ROC curve analysis indicated that the AUC value of the prediction model was 0.978(95％CI:0.945-0.994)in the training set and 0.959(95％CI:0.890-0.991)in the validation set.Decision curve analysis revealed that the model achieved higher net benefit when the threshold probability was between 20％and 80％.Conclusion Intracranial bleeding volume,GCS score,CRP,Didimer and FIB are related influencing factors of VTE in patients with acute stroke,and the prediction model of deci-sion curve based on the above factors is of high prediction efficiency.

The biological activity of chitinase in degrading chitin has garnered extensive attention,particularly for its potential applications in biological control.This study utilized four spore-form-ing Bacillus strains isolated from Dermacentor nuttalli ticks collected in the Hulunbuir region.Traditional bacterial culture methods were employed for isolation and identification,followed by 16S rRNA sequencing and phylogenetic analysis of the purified cultures.chitin-hydrolyzing strains were screened using colloidal chitin plates,and specific chitinase genes were detected via PCR.Fer-mentation was conducted at 37.0 ℃ for 4 d,and the supernatants were subjected to enzyme activity analysis using the DNS method.Four Gram-positive Bacillus strains were successfully isolated from tick tissue samples,they were identified as B.proteolyticus,B.paramycoides,B.thuringien-sis,and B.cereus,and renamed IMH/B-1,IMH/P-1,IMH/T-1,and IMH/C-1,respectively.PCR a-nalysis detected chitinase genes in B.proteolyticus and B.thuringiensis,while B.cereus and B.pa-ramycoides lacked these genes.However,three strains B.proteolyticus,B.thuringiensis,and B.ce-reus demonstrated significant(P＜0.01)chitin degradation activity on colloidal chitin.Enzyme ac-tivity assays revealed that chitinase activity ranged from 1.292 to 2.032 U/mL,with B.proteolytic-us exhibiting the highest activity 2.032 U/mL,followed by B.cereus 1.496 U/mL and B.thuring-iensis 1.324 U/mL.This study provides a foundation for further research and application of chiti-nase-producing Bacillus strains.

Nonunion of osteoporotic vertebral fractures (OVF), predominantly affecting the elderly, can lead to intractable pain, vertebral collapse, progressive kyphotic deformity, and neurological impairment, significantly compromising patients′ quality of life. There exists considerable debate on diagnosis and management of OVF, encompassing key issues such as clinical diagnosis and staging criteria for nonunion, surgical indications and procedure selection, and postoperative rehabilitation planning. Currently, there lacks standardized clinical guideline and expert consensus on the diagnosis and management of OVF nonunion in China. To address this gap, Minimally Invasive Surgery Group of Chinese Orthopedic Association, Osteoporosis Committee of Chinese Association of Orthopedic Surgeons, Prevention and Rehabilitation Committee for Osteoporosis of Chinese Association of Rehabilitation Medicine and Minimally Invasive Orthopedic Surgery Branch of China Association for Geriatric Care jointly organized domestic experts in spinal surgery, endocrinology, and rehabilitation to formulate the Clinical guideline for the diagnosis and treatment for nonunion of osteoporotic vertebral fractures ( version 2025), based on existing literature and clinical experience and adhering to principles of scientific rigor and practicality. The guideline provided 13 evidence-based recommendations encompassing diagnosis and treatment of OVF nonunion, aiming to standardize its clinical management.

Objective To construct a risk prediction model for venous thromboembolism(VTE)in patients with acute stroke and analyze its predictive value with decision curve.Methods Clinical data of 265 patients with acute stroke admitted to Department of Neurology of Pingxiang People's Hospital between May 2022 and May 2024 were collected,and 185 of them(69.8％)were assigned to a training set and 80 cases(30.2％)into a validation set.Based on the results of color Doppler ultrasound examination,the patients in the training set were divided into the VTE group(72 cases)and the non-VTE group(113 cases).Multivariate logistic regression analysis was used to identify the risk factors for VTE occurrence in the patients.A prediction model was constructed and a nomegram was developed to visually present the logistic regression prediction model.ROC curve and decision curve analyses were applied to evaluate the predictive performance of the logis-tic regression model.Results The VTE group had significantly larger intracranial hemorrhage volume,higher fibrinogen(FIB),D-dimer and C-reactive protein(CRP)levels,and lower Glasgow Coma Scale(GCS)score than the non-VTE group(P＜0.01).Multivariate logistic regression analysis indicated that intracranial hemorrhage volume(OR=1.243,95％CI:1.015-1.522),FIB(OR=3.161,95％CI:1.711-5.841),CRP(OR=1.595,95％CI:1.283-1.981),and D-dimer(OR=6.234,95％CI:2.693-14.434)were independent risk factors for the occurrence of VTE in the patients with acute stroke,while GCS score(OR=0.587,95％CI:0.413-0.834,P=0.003)was an independent protective factor against the occurrence.A VTE prediction model based on these influencing factors obtained a consistency index of 0.978.The calibration curve results showed that the observed values were in good agreement with the predicted values.ROC curve analysis indicated that the AUC value of the prediction model was 0.978(95％CI:0.945-0.994)in the training set and 0.959(95％CI:0.890-0.991)in the validation set.Decision curve analysis revealed that the model achieved higher net benefit when the threshold probability was between 20％and 80％.Conclusion Intracranial bleeding volume,GCS score,CRP,Didimer and FIB are related influencing factors of VTE in patients with acute stroke,and the prediction model of deci-sion curve based on the above factors is of high prediction efficiency.

Nonunion of osteoporotic vertebral fractures (OVF), predominantly affecting the elderly, can lead to intractable pain, vertebral collapse, progressive kyphotic deformity, and neurological impairment, significantly compromising patients′ quality of life. There exists considerable debate on diagnosis and management of OVF, encompassing key issues such as clinical diagnosis and staging criteria for nonunion, surgical indications and procedure selection, and postoperative rehabilitation planning. Currently, there lacks standardized clinical guideline and expert consensus on the diagnosis and management of OVF nonunion in China. To address this gap, Minimally Invasive Surgery Group of Chinese Orthopedic Association, Osteoporosis Committee of Chinese Association of Orthopedic Surgeons, Prevention and Rehabilitation Committee for Osteoporosis of Chinese Association of Rehabilitation Medicine and Minimally Invasive Orthopedic Surgery Branch of China Association for Geriatric Care jointly organized domestic experts in spinal surgery, endocrinology, and rehabilitation to formulate the Clinical guideline for the diagnosis and treatment for nonunion of osteoporotic vertebral fractures ( version 2025), based on existing literature and clinical experience and adhering to principles of scientific rigor and practicality. The guideline provided 13 evidence-based recommendations encompassing diagnosis and treatment of OVF nonunion, aiming to standardize its clinical management.

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.

Objective:To investigate the clinical features and prognosis of extranodal nasal-type NK/T-cell lymphoma of the extra-upper aerodigestive tract (extra-UADT NKTCL).Methods:The clinical data of 159 patients with extra-UADT NKTCL from the China Lymphoma Collaborative Group (CLCG) database between November 2001 and December 2015 were retrospectively analyzed. Kaplan-Meier survival analysis and Log-rank test were used to evaluate the prognosis. The Cox regression model is used for multi-factor analysis.Results:Extra-UADT NKTCL commonly occurs in skin and soft tissues (106/159, 66.7%) and gastrointestinal tract (31/159, 19.5%). The incidences of elevated lactate dehydrogenase (LDH) and Ann Arbor Ⅲ~Ⅳ stage were 47.8% (76/159) and 64.2% (102/159), respectively. The 3-year overall survival (OS) and progression-free survival (PFS) rates were 43.6% and 27.9%, respectively. The corresponding OS rates of primary skin/soft tissue site and gastrointestinal tract site were 41.0% and 59.4% ( P=0.281), while the PFS rates were 24.8% and 48.3%, respectively ( P=0.109). Combined modality treatment improved the 3-year OS of all the patients (58.4% vs 33.9%, P=0.001) and 3-year PFS (40.7% vs 20.7%, P=0.008) when compared with chemotherapy alone. LDH elevation, Ann Arbor synthesising and ≥2 junction external bits were intrusive as independent risk factors for total survival ( P<0.05), LDH elevation and ≥2 junction outer bits were intrusive as independent risk factors for progressionless survival( P<0.05). The distant extranodal dissemination was the primary failure patterns. Conclusions:Extra-UADT NKTCL appears to have distinct clinical characteristics and poor outcome. Compared with chemotherapy alone, combined modality treatment may improve the prognosis of patients with extra-UADT NKTCL.

Objective:To evaluate the prognosis and determine the failure patterns after radiotherapy for low-risk early-stage patients with extranodal NK/T-cell lymphoma, nasal-type (ENKTCL).Methods:A total of 557 patients from 2000—2015 with low-risk early-stage ENKTCL who received radiotherapy (RT) with or without chemotherapy (CT) from China Lymphoma Collaborative Group were retrospectively reviewed. Among them, 427 patients received combined modality therapy, whereas 130 patients received RT alone. Survivals were calculated by Kaplan-Meier method and compared with Log-rank test. Overall survival (OS) was compared with age and sex-matched general Chinese population using expected survival and standardized mortality ratio (SMR). Cox stepwise regression model was used for multivariate analysis.Results:The 5-year OS and progression-free survival (PFS) were 87.2% and 77.2%. The SMR was 3.59 ( P<0.001) at 1 year after treatment, whereas it was 1.50 at 4 years after treatment, without significant difference between ENKTCL group and country-matched general population ( P=0.146). Compared with RT alone, CMT did not result in significantly superior 5-year OS (87.0% vs 87.4%, P=0.961) or PFS (76.1% vs 80.7%, P=0.129). Local failure (11.5%, 64/557) and distant failure (10.8%, 60/557) were the main failure modes, while regional failure was rare (2.9%, 16/557). The 5-year locoregional control rate (LRC) was 87.2% for the whole group, with 89.5% for ≥50 Gy versus 73.7% for <50 Gy ( P<0.001). Radiotherapy dose was an independent factor affecting LRC( P<0.05). Conclusions:Radiotherapy achieves a favorable prognosis in patients with low-risk early-stage ENKTCL. The incidence of either locoregional or distant failure is low. Radiation dose still is an important prognostic factor for LRC.

Objective:To explore the effect of low frequency magnetic stimulation on myelin and inflammation in the callosum of demyelinated mice.Methods:Thirty-six 6 to 8-week-old male C57BL/6J mice were randomly divided into a control group, a cuprizone (CPZ) group and a magnetic therapy group. The CPZ group and the magnetic therapy group had demyelination induced by feeding a mixed diet containing 0.3% CPZ for 6 weeks, while the control group was given conventional food. The magnetic therapy group was given 50Hz 10mT magnetic stimulation during the 6 weeks for 20min daily, 5 days a week. The body mass of each mouse was observed every 7 days. At the end of the 6th week elevated cross maze experiments were conducted to observe any anxiety state. The myelin sheath in the corpus callosum was observed using Luxol fast blue staining and myelin basic protein (MBP) immunohistochemistry Tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in the corpus callosum were detected using enzyme-linked immunosorbent assays.Results:After the 6 weeks of treatment, the average body mass of the mice in the magnetic therapy group had improved significantly compared with the CPZ group. The CPZ group′s times in the elevated cross maze experiments were significantly shorter than those of the control group and also shorter than those of the magnetic therapy group. The Luxol staining showed significant myelin loss in the corpus callosum of the CPZ group, but compared with the CPZ group the average loss of myelin in the magnetic therapy group was significantly less. This was further confirmed by the MBP immunohistochemistry. Compared with the control group, the average expression of MBP in the CPZ group was significantly reduced, while in the magnetic therapy group it was significantly increased. Compared with the control group, the average TNF-α and IL-1β levels in the corpus callosum of the CPZ group increased significantly, but compared with the CPZ group the average levels in the magnetic therapy group had decreased significantly.Conclusions:Low frequency magnetic stimulation improves the body weight and anxiety state of mice. That is probably related to less myelin loss and inhibited inflammatory response in the corpus callosum.