OBJECTIVE To explore the clinical characteristics of the nontuberculous Mycobacteria pulmonary dis-ease(NTMPD)patients with previous pulmonary tuberculosis(PPTB)and analyze the clinical difference from the recurrence of pulmonary tuberculosis.METHODS By means of retrospective survey,the patients who were diag-nosed with NTMPD and recurrent pulmonary tuberculosis in Wuhan Pulmonary Hospital from Mar.2021 to Oct.2023 were recruited as the research subjects,a total of 395 patients with NTMPD were enrolled in the study and were divided into the PPTB-NTMPD group with 92 cases and the NPPTB-NTMPD group with 303 cases according to the history of PPTB.The baseline data,clinical symptoms,imaging findings,underlying diseases,pulmonary diseases,and species of nontuberculous Mycobacteria(NTM)were observed and compared.Totally 92 patients with recurrent pulmonary tuberculosis were randomly screened and assigned as the recurrent pulmonary tuberculo-sis group in a 1:1 ratio by matching the PPTB-NTMPD group with the gender and age.The major clinical charac-teristics were compared between the two groups.The 92 patients with PPTB-NTMPD were divided into the 1-10 years group with 40 cases,the 10-30 years group with 37 cases,and the more than 30 years group with 15 cases according to the interval between the initial diagnosis of pulmonary tuberculosis and the diagnosis of NTMPD.The major clinical characteristics were compared among the groups.RESULTS The age was(64.21±10.71)years old in the PPTB-NTMPD group,(60.26±11.83)years old in the NPPTB-NTMPD group(t=3.020,P=0.003).The proportion of patients with body mass index less than 18.5 kg/m2 was 59.78％in the PPTB-NTMPD group,41.25％in the NPPTB-NTMPD group(x2=6.155,P=0.013);the proportion of patients with cough was 77.17％in the PPTB-NTMPD group,65.68％in the NPPTB-NTMPD group(x2=4.313,P=0.038);the inci-dence of cavitary shadow was 50.00％in the PPTB-NTMPD group,35.31％in the NPPTB-NTMPD group(x2=6.414,P=0.011);the incidence of emphysema and pulmonary bullae was 29.35％in the PPTB-NTMPD group,12.87％in the NPPTB-NTMPD group(x2=13.766,P＜0.001);the incidence of chronic obstructive pulmonary disease(COPD)was 22.83％in the PPTB-NTMPD group,14.19％in the NPPTB-NTMPD group(x2=3.875,P=0.049);the incidence of damaged lung was 9.78％in the PPTB-NTMPD group,2.97％in the NPPTB-NT-MPD group(x2=7.530,P=0.014);there were significant differences.Mycobacterium intracellulare and Myco-bacterium abscessus were the predominant species of NTM in both the PPTB-NTMPD group and the NPPTB-NT-MPD group,there was no significant difference in the distribution of NTM species between the two groups of pa-tients.The incidence of patch shadow of the PPTB-NTMPD group was lower than that of the recurrent pulmonary tuberculosis group(P＜0.05),the incidence of bronchiectatic shadow of the PPTB-NTMPD group was higher than that of the recurrent pulmonary tuberculosis group(P＜0.05).There were significant differences in the age,incidence of pleural thickening and incidence of COPD among the patients with different time intervals between ini-tial diagnosis of pulmonary tuberculosis and the diagnosis of NTMPD in the PPTB-NTMPD group(P＜0.05).CONCLUSIONS The previous pulmonary tuberculosis mainly affect the body mass index less than 18.5 kg/m2 and the post-tuberculosis pulmonary diseases such as cough,pulmonary cavity,emphysema,pulmonary bullae,COPD and damaged lung of the NTMPD patients.The NTMPD patients with previous pulmonary tuberculosis are more likely to have bronchiectasia than the patients with recurrent tuberculosis.It is necessary for the clinicians to attach great importance.

Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration and hydroxylation.So far,few crystal structures of plant P450s have been obtained.We determined the crystal structure of IFS from Medicago truncatula at 1.9 ? by MAD method using a selenomethionine substituted crystal and conducted molecular docking and mutagenesis study.The structure of IFS complexed with imidazole exhibits the helix Ⅰa-loop-helix Ⅰβ motif which cor-responds to helix Ⅰ of other P450s.Compared with structures of common P450s,IFS/imidazole structure contains an extra domain,i.e.,the γ-domain.The structure reveals a homodimer in which the γ-domain of one molecule interacts with the β-domain of another.The plane of heme group makes an angle of ap-proximately 40° with the helix Ⅰa-loop-helix Ⅰβ motif.Molecular docking combined with mutagenesis study suggested that Trp-128 and Asp-300 might play important roles in substrate binding and recogni-tion.Phe-301,Ser-303 and Gly-305 from the helix Ⅰa-loop-helix Ⅰβ motif may play important roles in the aryl-ring migration.These novel structural features reveal insights into the unique reaction mechanism of IFS and provide a basis for engineering IFS in leguminous crops for health purpose.

To investigate the molecular mechanisms underlying the mechanosensitive ion channel PI-EZO2 in osteoarthritis(OA),we developed a lentiviral vector for endogenous PIEZO2 overexpression and established a stable PIEZO2-high-expressing immortalized human primary chondrocyte line.By map-ping the open reading frame of the PIEZO2 locus and designing sequence-specific sgRNA,we employed the CRISPR/Cas9 synergistic activation mediator(SAM)system to precisely integrate transcriptional ac-tivation elements into the PIEZO2 promoter region.Lentiviral-mediated targeted genomic integration en-sured endogenous PIEZO2 overexpression,confirmed by mCherry fluorescence tracing coupled with flow cytometric sorting,which revealed membrane-specific localization of PIEZO2 protein(localization effi-ciency:78.49％).Quantitative PCR demonstrated a 17-fold upregulation of PIEZO2 mRNA,while Western blotting validated enhanced membrane-localized protein expression.Strikingly,PIEZO2-overex-pressing chondrocytes exhibited hallmark OA metabolic phenotypes compared to wild-type controls:typeⅡ collagen mRNA expression decreased to 50％of baseline levels,whereas matrix metalloproteinase 13(MMP13)mRNA surged by 20-fold.These alterations recapitulated the pathological matrix metabolic phenotype observed in biomechanical OA models induced by cyclic mechanical stress(10％strain,0.5 Hz,8 h/day for 2 consecutive days).Collectively,we successfully generated a human chondrocyte model with stable PIEZO2 overexpression,which faithfully mirrors mechanotransduction-driven OA progression.This engineered cellular system provides a robust platform for dissecting PIEZO2-mediated mechanosig-naling networks and advancing targeted therapeutic discovery.

Objective To investigate the impacts of long non coding RNA(LncRNA)-PH domain containing 5 antisense RNA 1(FGD5-AS1)on the proliferation,migration,and invasion of colorectal cancer(CRC)cells by regulating the miR-299-5p/recombinant lysine specific demethylase 4B(KDM4B)axis.Methods The expressions of LncRNA FGD5-AS1,miR-299-5p,and KDM4B were detected by qRT-PCR in the surgically resected CRC tissues and adjacent tissues of 34 CRC patients CRC cells HCT116 were separated into si-NC group,si-FGD5-AS1 group,si-FGD5-AS1+inhibitor NC group,and si-FGD5-AS1+miR-299-5p inhibitor group.The levels of mRNA in each group were detected.CCK8,scratch assay,and Transwell assay were ap[plied to detect the proliferation,migration,and invasion of HCT116 cells in each group.Western blot was applied to detect the expression of proteins.Dual luciferase reporter gene assay verified the interaction mechanism between LncRNA FGD5-AS1 and miR-299-5p,and between miR-299-5p and KDM4B.Results LncRNA FGD5-AS1 and KDM4B were highly expressed in CRC tissue,while miR-299-5p was low expressed in CRC tissue(P＜0.05).The expression of LncRNA FGD5-AS1 mRNA,KDM4B mRNA,OD450,scratch healing rate,invasion number,PCNA,MMP-2,MMP-9,KDM4B in the si-FGD5-AS1 group were lower than those in the si-NC group,the expression of miR-299-5p mRNA was higher than that in the si-NC group(P＜0.05).Compared with the si-FGD5-AS1 group and si-FGD5-AS1+inhibitor NC group,the expression of miR-299-5p in the si-FGD5-AS1+miR-299-5p inhibitor group decreased(P＜0.05),the KDM4B mRNA,OD450,scratch healing rate,number of invasions,and expression of PCNA,MMP-2,MMP-9,KDM4B increased(P＜0.05).LncRNA FGD5-AS1 targeted negative regulation of miR-299-5p,while miR-299-5p targeted negative regulation of KDM4B.Conclusion Knocking down LncRNA FGD5-AS1 may inhibit the proliferation,migration,and invasion of CRC cells,and its mechanism may be achieved by regulating the miR-299-5p/KDM4B signaling axis.

Objective To investigate the impacts of long non coding RNA(LncRNA)-PH domain containing 5 antisense RNA 1(FGD5-AS1)on the proliferation,migration,and invasion of colorectal cancer(CRC)cells by regulating the miR-299-5p/recombinant lysine specific demethylase 4B(KDM4B)axis.Methods The expressions of LncRNA FGD5-AS1,miR-299-5p,and KDM4B were detected by qRT-PCR in the surgically resected CRC tissues and adjacent tissues of 34 CRC patients CRC cells HCT116 were separated into si-NC group,si-FGD5-AS1 group,si-FGD5-AS1+inhibitor NC group,and si-FGD5-AS1+miR-299-5p inhibitor group.The levels of mRNA in each group were detected.CCK8,scratch assay,and Transwell assay were ap[plied to detect the proliferation,migration,and invasion of HCT116 cells in each group.Western blot was applied to detect the expression of proteins.Dual luciferase reporter gene assay verified the interaction mechanism between LncRNA FGD5-AS1 and miR-299-5p,and between miR-299-5p and KDM4B.Results LncRNA FGD5-AS1 and KDM4B were highly expressed in CRC tissue,while miR-299-5p was low expressed in CRC tissue(P＜0.05).The expression of LncRNA FGD5-AS1 mRNA,KDM4B mRNA,OD450,scratch healing rate,invasion number,PCNA,MMP-2,MMP-9,KDM4B in the si-FGD5-AS1 group were lower than those in the si-NC group,the expression of miR-299-5p mRNA was higher than that in the si-NC group(P＜0.05).Compared with the si-FGD5-AS1 group and si-FGD5-AS1+inhibitor NC group,the expression of miR-299-5p in the si-FGD5-AS1+miR-299-5p inhibitor group decreased(P＜0.05),the KDM4B mRNA,OD450,scratch healing rate,number of invasions,and expression of PCNA,MMP-2,MMP-9,KDM4B increased(P＜0.05).LncRNA FGD5-AS1 targeted negative regulation of miR-299-5p,while miR-299-5p targeted negative regulation of KDM4B.Conclusion Knocking down LncRNA FGD5-AS1 may inhibit the proliferation,migration,and invasion of CRC cells,and its mechanism may be achieved by regulating the miR-299-5p/KDM4B signaling axis.

Objective:To integrate implicit templates with deep learning techniques, a novel neural network, the tooth-deformable deep implicit network (T-DDIN), was constructed to achieve high-precision shape completion of tooth defects in a personalized manner.Methods:A total of 550 intraoral scan models were collected from patients treated at the Department of Orthodontics and Department of Prosthodontics, West China Hospital of Stomatology, Sichuan University (500 for training and 50 for testing), between March 2022 and March 2024. T-DDIN reconstructed defective tooth morphology using an implicit template and a latent encoding prediction network. During model evaluation, Class Ⅱ cavity defects and occlusal wear defects were simulated in the test set. Morphological restoration was performed using both traditional computer aided design (CAD) methods and the T-DDIN deep learning approach. The two methods were compared based on three-dimensional deviation, occlusal adjustment volumes, cusp angle deviation, and restoration time.Results:The T-DDIN group demonstrated significantly lower three-dimensional deviation for Class Ⅱ cavity defects and occlusal wear restoration [(0.14±0.05) and (0.16±0.09) mm], occlusal adjustment volumes [(0.44±0.03) and (0.49±0.03) mm 3], and difference value of the tooth cusp angles (5.69°±1.90° and 6.04°±0.53°) compared to the traditional CAD group (both P<0.001). No significant differences were observed within the T-DDIN group between the two defect types in terms of three-dimensional deviation ( P=0.098) or occlusal adjustment volume ( P=0.154) or difference value of the tooth cusp angles ( P=0.196). However, in the traditional CAD group, three-dimensional deviation, occlusal adjustment volume and difference value of the tooth cusp angles was significantly higher in occlusal wear restorations than in Class Ⅱ cavity defects restorations ( P<0.001). The T-DDIN group, which involved Class Ⅱ cavity defects and occlusal wear, demonstrated significantly less recovery time of morphology (37.2±7.7) and (39.4±6.2) s compared to the traditional CAD group ( P<0.001). Conclusions:T-DDIN demonstrated superior stability and accuracy in morphological reconstruction for various types of dental defects while significantly reducing restoration time.

Objective:To explore the influence of initial high-risk cytogenetic abnormalities on the outcomes of children with acute myeloid leukemia (AML) after post-transplant Cyclophosphamide (PTCy)-based allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:A retrospective cohort study.AML children who underwent PTCy-based allo-HSCT after the first complete remission at Wuhan Children′s Hospital, Tongji Medical College, Huazhong University of Science and Technology between April 2017 and April 2024 were enrolled.Patients were divided into intermediate-risk and high-risk groups based on their initial cytogenetic features.These patients were further divided into complex karyotype, 11q23 rearrangement, and other karyotype groups.Clinical characteristics and survival outcomes were compared among these groups.Measurement and count data were analyzed using Wilcoxon rank-sum/Kruskal-Wallis and χ2 tests, respectively.Survival and risk factor analyses were performed using Kaplan-Meier and Cox proportional hazards methods, respectively. Results:A total of 51 AML children who underwent allo-HSCT were included in this study.The median age at transplantation was 3.2 years and the median follow-up time was 4.6 years.There were 26 cases in the intermediate-risk group and 25 cases in the high-risk group; 8 cases in the complex karyotype group, 14 cases in the 11q23 rearrangement group, and 29 cases in the other karyotype groups.By the end of the follow-up on November 30, 2024, 11 patients relapsed, 8 patients died, and 13 patients developed grades Ⅱ-Ⅳ acute graft-versus-host disease (GVHD).The 3-year overall survival (OS), relapse-free survival (RFS), and grades Ⅱ-Ⅳ acute GVHD-free and relapse-free survival (GRFS) were 84.0% (95% CI: 74.4%-94.8%), 74.5% (95% CI: 63.4%-87.5%), and 58.8% (95% CI: 46.7%-74.0%), respectively.The 3-year OS of the high-risk group was significantly lower than that of the intermediate-risk group (71.8% vs.96.2%, P=0.022), while differences in 3-year RFS and GRFS between the 2 groups were not statistically significant (68.0% vs.80.8%, P=0.400; 52.0% vs.65.4%, P=0.420).The 3-year OS, RFS and GRFS of the complex karyotype group were significantly lower than those of 11q23 rearrangement and other karyotype groups (50.0% vs.85.7%, 93.1%, P=0.009; 37.5% vs.85.7%, 79.3%, P=0.022; 25.0% vs.64.3%, 65.5%, P=0.049).Multivariate analysis showed that a complex karyotype was an independent prognostic factor affecting 3-year OS and GRFS [OS: HR=6.79 (95% CI: 1.13-43.80), P=0.044; GRFS: HR=3.72(95% CI: 1.13-12.20), P=0.030]. Conclusions:High-risk cytogenetic features are significant predictors of survival outcomes in pediatric AML patients undergoing PTCy-based allo-HSCT.

Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration and hydroxylation.So far,few crystal structures of plant P450s have been obtained.We determined the crystal structure of IFS from Medicago truncatula at 1.9 ? by MAD method using a selenomethionine substituted crystal and conducted molecular docking and mutagenesis study.The structure of IFS complexed with imidazole exhibits the helix Ⅰa-loop-helix Ⅰβ motif which cor-responds to helix Ⅰ of other P450s.Compared with structures of common P450s,IFS/imidazole structure contains an extra domain,i.e.,the γ-domain.The structure reveals a homodimer in which the γ-domain of one molecule interacts with the β-domain of another.The plane of heme group makes an angle of ap-proximately 40° with the helix Ⅰa-loop-helix Ⅰβ motif.Molecular docking combined with mutagenesis study suggested that Trp-128 and Asp-300 might play important roles in substrate binding and recogni-tion.Phe-301,Ser-303 and Gly-305 from the helix Ⅰa-loop-helix Ⅰβ motif may play important roles in the aryl-ring migration.These novel structural features reveal insights into the unique reaction mechanism of IFS and provide a basis for engineering IFS in leguminous crops for health purpose.

Objective:To explore the influence of initial high-risk cytogenetic abnormalities on the outcomes of children with acute myeloid leukemia (AML) after post-transplant Cyclophosphamide (PTCy)-based allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:A retrospective cohort study.AML children who underwent PTCy-based allo-HSCT after the first complete remission at Wuhan Children′s Hospital, Tongji Medical College, Huazhong University of Science and Technology between April 2017 and April 2024 were enrolled.Patients were divided into intermediate-risk and high-risk groups based on their initial cytogenetic features.These patients were further divided into complex karyotype, 11q23 rearrangement, and other karyotype groups.Clinical characteristics and survival outcomes were compared among these groups.Measurement and count data were analyzed using Wilcoxon rank-sum/Kruskal-Wallis and χ2 tests, respectively.Survival and risk factor analyses were performed using Kaplan-Meier and Cox proportional hazards methods, respectively. Results:A total of 51 AML children who underwent allo-HSCT were included in this study.The median age at transplantation was 3.2 years and the median follow-up time was 4.6 years.There were 26 cases in the intermediate-risk group and 25 cases in the high-risk group; 8 cases in the complex karyotype group, 14 cases in the 11q23 rearrangement group, and 29 cases in the other karyotype groups.By the end of the follow-up on November 30, 2024, 11 patients relapsed, 8 patients died, and 13 patients developed grades Ⅱ-Ⅳ acute graft-versus-host disease (GVHD).The 3-year overall survival (OS), relapse-free survival (RFS), and grades Ⅱ-Ⅳ acute GVHD-free and relapse-free survival (GRFS) were 84.0% (95% CI: 74.4%-94.8%), 74.5% (95% CI: 63.4%-87.5%), and 58.8% (95% CI: 46.7%-74.0%), respectively.The 3-year OS of the high-risk group was significantly lower than that of the intermediate-risk group (71.8% vs.96.2%, P=0.022), while differences in 3-year RFS and GRFS between the 2 groups were not statistically significant (68.0% vs.80.8%, P=0.400; 52.0% vs.65.4%, P=0.420).The 3-year OS, RFS and GRFS of the complex karyotype group were significantly lower than those of 11q23 rearrangement and other karyotype groups (50.0% vs.85.7%, 93.1%, P=0.009; 37.5% vs.85.7%, 79.3%, P=0.022; 25.0% vs.64.3%, 65.5%, P=0.049).Multivariate analysis showed that a complex karyotype was an independent prognostic factor affecting 3-year OS and GRFS [OS: HR=6.79 (95% CI: 1.13-43.80), P=0.044; GRFS: HR=3.72(95% CI: 1.13-12.20), P=0.030]. Conclusions:High-risk cytogenetic features are significant predictors of survival outcomes in pediatric AML patients undergoing PTCy-based allo-HSCT.

OBJECTIVE: To explore the efficacy and safety of human epidermal growth factor gel in the treatment of pin tract infections after surgery in patients with severe spinal deformity. METHODS: A retrospective case-control study was conducted to analyze the clinical data of 26 patients with pin tract infections after skull-pelvic ring traction for severe spinal deformity admitted from February 2019 to May 2022. Among them, 11 were male and 15 were female;the age ranged from 18 to 31 years, with an average of (24.1±4.3) years;the Cobb angle ranged 80.3° to 120.7°, with an average of (88.6±10.2)°;there were 52 iliac traction pins, 104 pin tract openings, and 38 pin tract infections. According to the Checketts-Otterbum classification, there were 11 infections of gradeⅠ, 13 infections of gradeⅡ, 10 infections of grade Ⅲ, and 4 infections of grade Ⅳ. The patients were divided into the conventional dressing change group (13 cases) and the growth factor group (13 cases) by coin flipping. Clinical efficacy was evaluated by recording the visual analogue scale(VAS) score during dressing change, duration of dressing change, cost of dressing change, positive rate of bacterial culture, wound improvement rate, and wound improvement time. RESULTS: There were no statistically significant differences in VAS or duration of dressing change between the two groups (P>0.05). The cost of dressing change was (800.0±59.5) yuan in the conventional dressing change group and (1 179.5±80.9) yuan in the growth factor group, with a statistically significant difference (P<0.05). There was no statistically significant difference in the positive rate of bacterial culture between the two groups (P>0.05). In the conventional dressing change group, within 20 pin tract infections, 2 infections achieved wound healing, 7 infections showed improvement, and 11 infections were ineffective;in the growth factor group, within 18 pin tract infections 5 infections achieved wound healing, 8 infections showed improvement, and 5 infections were ineffective, with a statistically significant difference between the two groups (P<0.05). The wound healing time was (22.8±4.9) days in the conventional dressing change group and (14.2±2.5) days in the growth factor group, with a statistically significant difference (P<0.05). No complications occurred in either group. CONCLUSION The application of human epidermal growth factor gel in the treatment of pin tract infections after skull-pelvic ring surgery in patients with severe spinal deformity is easy to operate, does not increase patient pain, and has significant efficacy. It shortens wound healing time, effectively promotes wound healing, and has good safety and high cost-effectiveness.
Humans ; Male ; Female ; Adult ; Adolescent ; Retrospective Studies ; Case-Control Studies ; Young Adult ; Epidermal Growth Factor/therapeutic use* ; Traction/adverse effects* ; Gels ; Bone Nails/adverse effects* ; Surgical Wound Infection/drug therapy*