Glucose oxidase (GOD) is an oxygen-consuming dehydrogenase that can catalyze the production of gluconic acid hydrogen peroxide from glucose, and its specific mechanism of action makes it promising for applications, while the low catalytic activity and poor thermostability have become the main factors limiting the industrial application of this enzyme. In this study, we used the glucose oxidase AtGOD reported with the best thermostability as the source sequence for phylogenetic analysis to obtain the GOD with excellent performance. Six genes were screened and successfully synthesized for functional validation. Among them, the glucose oxidase AhGODB derived from Aspergillus heteromorphus was expressed in Pichia pastoris and showed better thermostability and catalytic activity, with an optimal temperature of 40 ℃, a specific activity of 112.2 U/mg, and a relative activity of 47% after 5 min of treatment at 70 ℃. To improve its activity and thermal stability, we constructed several mutants by directed evolution combined with rational design. Compared with the original enzyme, the mutant T72R/A153P showcased the optimum temperature increasing from 40 to 50 ℃, the specific activity increasing from 112.2 U/mg to 166.1 U/mg, and the relative activity after treatment at 70 ℃ for 30 min increasing from 0% to 33%. In conclusion, the glucose oxidase mutants obtained in this study have improved catalytic activity and thermostability, and have potential for application.
Glucose Oxidase/chemistry* ; Enzyme Stability ; Aspergillus/genetics* ; Pichia/metabolism* ; Temperature ; Catalysis ; Fungal Proteins/metabolism* ; Hot Temperature

Objective:This study aims to evaluate the nasal structural and electrophysiological features of patients with postoperative olfactory dysfunction following aesthetic rhinoplasty.Methods:We retrospectively analyzed the clinical features of 30 outpatients (females, aged 33±6 years) from Beijing Anzhen Hospital and China-Japan Friendship Hospital between 2014 and 2023, who complained of olfactory dysfunction following aesthetic rhinoplasty. The control group was 30 healthy females aged 32±9 years. Psychophysical olfactory test (Sniffin′ Sticks, SS), olfactory and trigeminal event-related potentials (oERPs and tERPs), and acoustic rhinometry were used for evaluating the olfactory function and nasal structure in patients and healthy controls. SPSS 17.0 software was used to compare the difference in olfactory function and nasal structure between the two groups and to analyze the factors related postoperative olfactory dysfunction.Results:There was a significant difference in the scores on psychophysical olfactory test between the patients and controls (10.78±3.90 vs. 33.66±2.42, t=-23.35, P<0.001). ERPs could be evoked in all patients and controls. Patients showed higher amplitudes of N 1 waves in both oERPs and tERPs than controls ( P<0.05 for all), but no differences in the latencies of N 1 and P 2 waves or in the amplitudes of P 2 waves were observed between the two groups ( P>0.05 for all). There was no difference in nasal structure between the two groups ( P>0.05). However, after nasal decongestant, mucosal congestion in the cross-sectional area (CSA) from the nostril to 6 cm level was found more significantly in patients than controls (nasal congestion index 40.00% vs. 1.00%, t=2.09, P=0.047). Better olfactory function was associated with increasing nasal volumes, increasing nasal threshold and anterior nasal turbinate plane CSA( P<0.05 for all). Conclusion:The important factor related to olfactory dysfunction following aesthetic rhinoplasty may be attributed to local mucosal congestion, rather than nasal structural alteration or neurophysiologic deficits in the olfactory pathway.

Objective:To evaluate the effectiveness and safety of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in children with stage 4/M neuroblastoma (NB).Methods:This study was a prospective, single-arm, multicenter clinical trial conducted by Sun Yat-sen Memorial Hospital, Children′s Hospital of Nanjing Medical University, Children′s Hospital, Zhejiang University School of Medicine, the First Affiliated Hospital of Guangxi Medical University, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine. From March, 2019 to August, 2023, 25 children with confirmed with stage 4/M NB and received allo-HSCT were enrolled. The patients received either unrelated cord blood transplantation (UCBT) or peripheral blood stem cell transplantation (PBSCT). Conditioning regimens for UCBT was fludarabine+busulfan+cyclophosphamide+topotecan, and for PBSCT was fludarabine+busulfan+melphalan+thiotepa+antithymocyte globulin, respectively. Until the last follow-up date of September, 2023, the overall survival (OS) rate and event free survival (EFS) rate were analyzed to evaluate efficacy. The engraftment rate and transplant-related complications were statistically assessed to evaluate safety. Survival analysis was performed using the Kaplan-Meier method.Results:Of the 25 patients, there were 15 males and 10 females. The age at transplantation was 5.7 (3.8, 7.3) years. The engraft rate was 100%, with recovery time of neutrophil as 15.7 (12.5, 17.0) d, and the recovery time of platelets as 33.5 (18.0, 48.0) d. Seventeen of the 25 children (68%) developed acute graft versus host disease (aGVHD), occurred at 18.0 (13.0, 22.5) d after transplantation, including 13 of grade Ⅲ-Ⅳ cases. The main sites of aGVHD were skin and intestinal tract. After treatment, 13 cases improved, 4 patients developed chronic graft-versus-host disease (cGVHD). After allo-HSCT, 14 children received maintenance therapy. Twenty of the 25 patients survived, the 2-year cumulative OS rate was (80±9)%, and 2-year EFS rate was (56±11)%. Nine cases (36%) relapsed, the time from allo-HSCT to disease relapse was 10.9 (5.5, 16.0) months. Five cases (20%) died. The hematopoietic stem cell transplantation associated mortality rate was 4% (1/25).The 2-year OS rate of patients who had partial remission prior to allo-HSCT was significant lower than those who had complete remission prior to allo-HSCT ((33±25)% vs. 100%, P=0.037). Conclusion:allo-HSCT is an effective treatment for patients with stage 4/M NB.

Objective:To investigate the efficacy and safety of allogeneic hematopoietic stem cell transplantation (allo-HSCT) for the treatment of BCR::ABL-negative chronic neutrophilic leukemia (CNL) and MDS/MPN with neutrophilia.Methods:This study retrospectively analyzed 12 cases of CNL and MDS/MPN with neutrophilia that underwent allo-HSCT from March 2017 to June 2024, comprising 7 males and 5 females with a median age of 48 ( IQR: 28, 59) years. The 2-year overall survival (OS), disease-free survival (DFS), cumulative incidence of relapse (CIR), and transplantation-related mortality (TRM) rates were analyzed. Complications were also assessed. Results:Of the 12 patients, 6 received matched sibling HSCT and 6 received haploidentical HSCT. All patients had successful engraftment, and the median times of neutrophil and platelet engraftment were 17 ( IQR: 11, 24) days and 15 ( IQR: 9, 28) days, respectively. Grade Ⅱ–Ⅳ acute graft versus host disease (GVHD) and chronic GVHD occurred in 2 and 4 cases, respectively. The 2-year OS, DFS, CIR, and TRM rates were (65.6 ± 16.4) %, (41.7 ± 16.6) %, (47.2 ±18.2) %, and (11.1 ± 11.4) %, respectively, after a median follow-up time of 637 ( IQR: 330, 943) days. One patient died from treatment-related complications due to respiratory failure caused by coronavirus disease 2019. Two patients died due to relapse. Conclusion:Allo-HSCT can be applied as a safe and effective approach to treat CNL and MDS/MPN with neutrophilia.

Objective:To evaluate the safety and efficacy of donor-purified CD34 + stem cell boosts in patients with poor hematopoietic reconstruction (PHR) after haploid hematopoietic stem cell transplantation (haplo-HSCT) for aplastic anemia (AA) . Method:A retrospective analysis was conducted on 11 patients with AA and PHR who underwent haplo-HSCT and received donor-purified CD34 + stem cell boosts at Peking University People’s Hospital. Recovery of blood cell counts, incidence of graft-versus-host disease (GVHD), and overall survival (OS) were assessed. Results:Of the 11 patients with PHR, two were diagnosed with prolonged isolated thrombocytopenia (PT), one was primary poor graft function (PGF), and eight were diagnosed with secondary PGF. The median time to PHR diagnosis was 110 days (range: 60-330 days), and the median interval from transplantation to purified CD34 + hematopoietic stem cell infusion was 194 days (range: 125-456 days). The two patients with PT achieved complete platelet recovery at 22 and 13 days after CD34 + stem cell infusion, respectively. Among the remaining nine patients with PGF, six achieved complete hematopoietic recovery, with a median absolute neutrophil count recovery time of 19 days (8-158 days), HGB recovery time of 32.5 days (range: 13-158 days), and platelet recovery time of 31.5 days (range: 7-171 days). The incidence of chronic GVHD after infusion was 18.2%, with no cases of acute GVHD observed. The OS rate was 90.9% (10/11) in the 11 patients, with a median follow-up of 614 days (range: 153-1 765 days) . Conclusion:Donor-purified CD34 + stem cell boost may be an effective therapeutic strategy for PHR in patients with AA after haplo-HSCT.

Objective:To evaluate the diagnostic value of targeted next-generation sequencing (tNGS) of throat swab samples for detecting community-acquired respiratory viruses (CARV) in patients with hematological diseases.Methods:Clinical and laboratory data from 64 episodes involving patients with hematological diseases and suspected infections—who underwent both pharyngeal swab tNGS and CARV polymerase chain reaction (PCR) testing concurrently—were retrospectively analyzed. The cases were drawn from the Department of Hematology, Peking University People’s Hospital, between September 2023 and April 2024. Concordance between tNGS and CARV PCR results, as well as the diagnostic performance of tNGS in detecting CARV, were evaluated.Results:Among the 64 episodes, 29 were clinically diagnosed with respiratory tract infections, including one case of cytomegalovirus pneumonia and 28 CARV-positive cases. The remaining 35 episodes involved patients with fever or respiratory symptoms attributed to other causes, including 14 with extrapulmonary infections and 21 with noninfectious etiologies. The median follow-up duration was 215.5 days (range: 7-271 days). PCR detected 24 strains of seven CARV types, whereas tNGS detected 25 strains of eight CARV types. Using PCR results as the reference standard, the sensitivity, specificity, positive predictive value, negative predictive value and accuracy of tNGS were 85.0%, 88.6%, 77.3%, 92.9%, and 87.5%, respectively. The two methods showed good concordance (Kappa=0.717, P<0.001) . Conclusion:Pharyngeal swab tNGS may serve as a viable alternative to PCR for diagnosing CARV infections in patients with hematological diseases.

Objective:To investigate the inhibitory effect of Huqizhengxiao decoction (HQZXD) on subcutaneous tumor in H22 hepatoma-bearing mice and its potential mechanism.Methods:Twenty-five healthy male BALB/c inbred mice aged 4 to 6 weeks and weighing (20±2) g were taken. One of them was used for the amplification of H22 hepatoma cells. The amplified H22 hepatoma cells were inoculated subcutaneously at the left posterior axillary line of the remaining mice for modeling. After subcutaneous tumor formation, the mice were randomly divided into four groups: model group, HQZXD group, sorafenib group and combined (HQZXD+ sorafenib) group, with 6 mice in each group. Tumor inhibition rates, and serum levels of aspartate transaminase (AST) and alanine transaminase (ALT) were observed. The expression of interleukin (IL)-6, signal transducer and activator of transcription 3 (STAT3), phosphorylated STAT3 (p-STAT3), and nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) in tumor tissues was detected using immunohistochemistry and Western blotting. Enzyme-linked immunosorbent assay was used to quantify the levels of IL-6, tumor necrosis factor-alpha (TNF-α), and IL-1β in tumor tissues. Quantitative real-time polymerase chain reaction (qRT-PCR) was employed to assess the mRNA levels of IL-6, STAT3, and C-X-C motif chemokine ligand 1 (CXCL1) in tumor tissues.Results:The general condition of mice in all treatment groups improved compared to the model group. Notably, the tumor weight (0.50±0.22) g and tumor volume (0.37±0.18) cm 3 in the combined group were significantly lower than those in the model group [tumor weight: (1.63±0.26) g, tumor volume: (0.98±0.83) cm 3] with statistical significance (both P<0.05). The tumor inhibition rates for the sorafenib, HQZXD, and combination groups were 35.4%, 48.6%, and 69.7%, respectively. Compared to the model group, serum levels of AST and ALT were reduced in all treatment groups, with the combined group showing the most significant decrease [AST: (48.81±2.82) U/L vs. (188.12±6.51) U/L; ALT: (34.14±1.25) U/L vs. (116.62±4.72) U/L], and the differences were statistically significant (both P<0.05). The protein expression levels of IL-6, STAT3, p-STAT3, IL-1β, TNF-α, and NLRP3 in tumor tissues were reduced in all treatment groups compared to the model group, with the combined group showing the most marked reduction, and the differences were statistically significant (all P<0.05). Similarly, the mRNA levels of IL-6, STAT3, and CXCL1 in tumor tissues were lower in all treatment groups compared to the model group, with the combined group showing lower levels than the single treatment groups, and these differences were statistically significant (all P<0.05). Conclusion:HQZXD can inhibit the activation of IL-6/STAT3 pathway, reduce inflammation in tumors, and consequently play a certain inhibitory effect on tumor.

Objective To investigate the efficacy of combined telephone and WeChat follow-up for patients discharged with an indwelling closed thoracic drainage tube after surgery for stage Ⅲ tuberculous empyema. Methods Patients with stage Ⅲ tuberculous empyema who were discharged with an indwelling drainage tube from the Department of Thoracic Surgery, Public Health Clinical Center of Chengdu, between November 2021 and November 2022 were enrolled in this study. They were divided into an observation group (combined telephone and WeChat follow-up) and a control group (telephone-only follow-up). The quality of life (QoL), treatment adherence, and recovery outcomes were compared between the two groups. Results A total of 81 patients were included. The observation group consisted of 49 patients (31 males, 18 females) with a mean age of (38.63±15.86) years. The control group consisted of 32 patients (27 males, 5 females) with a mean age of (36.91±17.33) years. The observation group showed significantly better postoperative QoL outcomes in the domains of physical functioning, emotional functioning, physical symptoms, global health status, and overall QoL compared to the control group (all P<0.05). Regarding treatment adherence, the observation group demonstrated superior performance in daily activity duration, use of a respiratory trainer, and adherence to coughing exercises compared to the control group (all P<0.001). The duration of chest tube indwelling was significantly shorter in the observation group (P<0.001). Furthermore, the observation group showed better recovery in albumin and hemoglobin levels (both P<0.05). Conclusion A combined telephone and WeChat follow-up approach can significantly improve the QoL and treatment adherence for patients discharged with an indwelling drainage tube. This method effectively shortens the duration of postoperative tube drainage, promotes nutritional recovery, and accelerates overall postoperative rehabilitation.

Accurate prediction of molecular properties is crucial for selecting compounds with ideal properties and reducing the costs and risks of trials.Traditional methods based on manually crafted features and graph-based methods have shown promising results in molecular property prediction.However,traditional methods rely on expert knowledge and often fail to capture the complex structures and interactions within molecules.Similarly,graph-based methods typically overlook the chemical structure and function hidden in molecular motifs and struggle to effectively integrate global and local molecular information.To address these limitations,we propose a novel fingerprint-enhanced hierarchical graph neural network(FH-GNN)for molecular property prediction that simultaneously learns information from hierarchical molecular graphs and fingerprints.The FH-GNN captures diverse hierarchical chemical information by applying directed message-passing neural networks(D-MPNN)on a hierarchical molecular graph that integrates atomic-level,motif-level,and graph-level information along with their relationships.Addi-tionally,we used an adaptive attention mechanism to balance the importance of hierarchical graphs and fingerprint features,creating a comprehensive molecular embedding that integrated hierarchical mo-lecular structures with domain knowledge.Experiments on eight benchmark datasets from MoleculeNet showed that FH-GNN outperformed the baseline models in both classification and regression tasks for molecular property prediction,validating its capability to comprehensively capture molecular informa-tion.By integrating molecular structure and chemical knowledge,FH-GNN provides a powerful tool for the accurate prediction of molecular properties and aids in the discovery of potential drug candidates.

HDAC7, a member of class IIa HDACs, plays a pivotal regulatory role in tumor, immune, fibrosis, and angiogenesis, rendering it a potential therapeutic target. Nevertheless, due to the high similarity in the enzyme active sites of class IIa HDACs, inhibitors encounter challenges in discerning differences among them. Furthermore, the substitution of key residue in the active pocket of class IIa HDACs renders them pseudo-enzymes, leading to a limited impact of enzymatic inhibitors on their function. In this study, proteolysis targeting chimera (PROTAC) technology was employed to develop HDAC7 drugs. We developed an exceedingly selective HDAC7 PROTAC degrader B14 which showcased superior inhibitory effects on cell proliferation compared to TMP269 in various diffuse large B cell lymphoma (DLBCL) and acute myeloid leukemia (AML) cells. Subsequent investigations unveiled that B14 disrupts BCL6 forming a transcriptional inhibition complex by degrading HDAC7, thereby exerting proliferative inhibition in DLBCL. Our study broadened the understanding of the non-enzymatic functions of HDAC7 and underscored the importance of HDAC7 in the treatment of hematologic malignancies, particularly in DLBCL and AML.