Objective:To identify strain-specific features of Klebsiella pneumoniae by analyzing metagenomics next generation sequencing (mNGS) data, thereby expanding the downstream applications of mNGS. Methods:The sequences of K.pneumoniae strains were organized from both the self-built database of the long-term multi-center research cohort in China established by the Peking University People′s Hospital from 2009 to 2020 (with 2 345 sequences) and the public databases (with 19 648 sequences). The existing large-scale databases were compressed, and a set of strains representative of clonal groups were screened. A strain genome information library was constructed based on k-mer features, and the most matching representative sequences in the database were searched for the raw mNGS data. The search results of the self-built library and public library were merged and optimized to update the prediction of antimicrobial-resistance characteristics and avoid the impact of uneven data distribution on the results. A total of 314 clinical samples from patients with K.pneumoniae detected by mNGS in the Clinical Microbiology Laboratory of Peking University People′s Hospital from 2022 to 2024 were retrospectively collected, and 101 samples with positive clinical culture results were selected to validate the prediction results. The antimicrobial-resistance phenotypes were verified by clinical antimicrobial susceptibility test results. Whole-genome sequencing was performed on the culture strains of 14 samples randomly selected using random numbers to verify the genotypes. Single nucleotide polymorphism distance analysis was used to verify the occurrence of outbreak events. The χ2 test and Mann-Whitney U test were used for statistical analysis. Results:A representative strain sequence k-mer feature library containing self-built and public sub-libraries was constructed. The library construction required only about 1 hour with <3 GB storage, with a high compression ratio and low update cost. Using k-mer-based analysis, mNGS data achieved precise strain characterization within 4 minutes and and <5 GB memory occupation. There was a significant difference in the antimicrobial-resistance rates to more than half of the antibiotics between the self-built database (90.8%, 2 130/2 345) and the public database (22.7%, 4 457/19 648) ( χ2=4 634.1, P<0.001). After optimizing the search results, the mean category agreement, sensitivity, and specificity of the prediction for eight antibiotics reached 84.8% (323/381), 78.9% (131/166), and 91.2% (196/215), respectively. The target genotypes were successfully detected in 10 out of 12 samples, and two outbreak events (2 samples per event) were successfully identified. Conclusions:An independent analysis process adapted to the needs of identifying the features of K. pneumoniae strains in mNGS data was developed. This process requires minimal computational resources and processing time and can directly achieve the simultaneous analysis of the antimicrobial-resistance phenotypes of K. pneumoniae at the strain level and their corresponding genomic characteristic profiles based on the raw mNGS reads.

High mobility group box 1 (HMGB1), when released extracellularly, plays a pivotal role in the development of spinal cord synapses and exacerbates autoimmune diseases within the central nervous system. In experimental autoimmune encephalomyelitis (EAE), a condition that models multiple sclerosis, the levels of extracellular HMGB1 and interleukin-33 (IL-33) have been found to be inversely correlated. However, the mechanism by which IL-33 deficiency enhances HMGB1 release during EAE remains elusive. Our study elucidates a potential signaling pathway whereby the absence of IL-33 leads to increased binding of P300/CBP-associated factor with HMGB1 in the nuclei of astrocytes, upregulating HMGB1 acetylation and promoting its release from astrocyte nuclei in the spinal cord of EAE mice. Conversely, the addition of IL-33 counteracts the TNF-α-induced increase in HMGB1 and acetylated HMGB1 levels in primary astrocytes. These findings underscore the potential of IL-33-associated signaling pathways as a therapeutic target for EAE treatment.
Animals ; Encephalomyelitis, Autoimmune, Experimental/metabolism* ; Astrocytes/metabolism* ; Interleukin-33/metabolism* ; HMGB1 Protein/metabolism* ; Acetylation ; Mice, Knockout ; Mice, Inbred C57BL ; p300-CBP Transcription Factors/metabolism* ; Mice ; Spinal Cord/metabolism* ; Cells, Cultured ; Female ; Signal Transduction

ObjectiveTo explore the dose-effect relationship and safety of high， medium， and low doses of raw Astragali Radix in the modified Huangqi Chifengtang （MHCD） for treating proteinuria in immunoglobulin A （IgA） nephropathy， and to provide scientific evidence for the clinical use of high-dose Astragali Radix in the treatment of proteinuria in IgA nephropathy. MethodsA total of 120 patients with IgA nephropathy， diagnosed with Qi deficiency and blood stasis combined with wind pathogen and heat toxicity， were randomly divided into a control group and three treatment groups. The control group received telmisartan combined with a Chinese medicine placebo， while the treatment groups were given telmisartan combined with MHCD containing different doses of raw Astragali Radix （60， 30， 15 g）. Each group contained 30 patients， and the treatment period was 12 weeks. Changes in 24-hour urinary protein （24 hUTP）， traditional Chinese medicine （TCM） syndrome scores， effective rate， and renal function were observed before and after treatment. Safety was assessed by monitoring liver function and blood routine. ResultsAfter 12 weeks of treatment， 24 hUTP significantly decreased in the high， medium， and low-dose groups， as well as the control group （P<0.05， P<0.01）. The TCM syndrome scores in the high， medium， and low-dose groups also significantly decreased （P<0.01）. Comparisons between groups showed that the 24 hUTP in the high-dose group was significantly lower than in the medium， low-dose， and control groups （P<0.05， P<0.01）， and the 24 hUTP in the medium-dose group was significantly lower than in the control group （P<0.05）. The TCM syndrome scores in the high and medium-dose groups were significantly lower than in the low-dose and control groups （P<0.05， P<0.01）. The total effective rates for proteinuria in the high， medium， low-dose， and control groups were 92.59% （25/27）， 85.19% （23/27）， 60.71% （17/28）， and 57.14% （16/28）， respectively. The effective rates in the high and medium-dose groups were significantly higher than in the low-dose and control groups （χ²=13.185， P<0.05， P<0.01）. The effective rates for TCM syndrome scores in the high， medium， low-dose， and control groups were 88.89% （24/27）， 81.48% （22/27）， 71.43% （20/28）， and 46.43% （13/28）， respectively. The efficacy of TCM syndrome scores in the high and medium-dose groups was significantly higher than in the control group （χ²=14.053， P<0.01）. Compared with pre-treatment values， there was no statistically significant difference in eGFR and serum creatinine in the high and medium-dose groups. However， eGFR significantly decreased in the low-dose and control groups after treatment （P<0.05）， and serum creatinine levels increased significantly in the control group （P<0.05）. No statistically significant differences were observed in urea nitrogen， uric acid， albumin， total cholesterol， triglycerides， liver function， and blood routine before and after treatment in any group. ConclusionThere is a dose-effect relationship in the treatment of IgA nephropathy with high， medium， and low doses of raw Astragali Radix in MHCD. The high-dose group exhibited the best therapeutic effect and good safety profile.

The cooccurrence of NPM1, FLT3-ITD, and DNMT3A mutations (i.e., triple mutation) is related to dismal prognosis in patients with acute myeloid leukemia (AML) receiving chemotherapy alone. In this multicenter retrospective cohort study, we aimed to identify whether allogeneic hematopoietic stem cell transplantation (allo-HSCT) could overcome the poor prognosis of DNMT3A^mutNPM1^mutFLT3-ITD^mut AML across four transplant centers in China. Fifty-three patients with triple-mutated AML receiving allo-HSCT in complete remission were enrolled. The 1.5-year probabilities of relapse, leukemia-free survival, and overall survival after allo-HSCT were 11.9%, 80.3%, and 81.8%, respectively. Multivariate analysis revealed that more than one course of induction chemotherapy and allo-HSCT beyond CR1 were associated with poor survival. To our knowledge, this work is the largest study to explore the up-to-date undefined role of allo-HSCT in patients with triple-mutated AML. Our real-world data suggest that allo-HSCT could overcome the poor prognosis of DNMT3A^mutNPM1^mutFLT3-ITD^mut in AML.
Humans ; Nucleophosmin ; Leukemia, Myeloid, Acute/mortality* ; Hematopoietic Stem Cell Transplantation/methods* ; Male ; Female ; DNA Methyltransferase 3A ; Adult ; China ; Retrospective Studies ; DNA (Cytosine-5-)-Methyltransferases/genetics* ; Middle Aged ; Prognosis ; fms-Like Tyrosine Kinase 3/genetics* ; Mutation ; Young Adult ; Transplantation, Homologous ; Nuclear Proteins/genetics* ; Adolescent ; Aged

OBJECTIVES: The application of photodynamic therapy in solid tumors has attracted increasing attention in recent years, and the efficiency of photosensitizers is a crucial determinant of therapeutic efficacy. This study aims to evaluate the cytotoxic effects of a novel photosensitizer, PEG-MTPABZ-PyC, in photodynamic therapy against gastric cancer cells. METHODS: Gastric cancer MKN45 cells were treated with PEG-MTPABZ-PyC. A high-content live-cell imaging system was used to assess the cellular uptake kinetics and subcellular localization of the photosensitizer. The cytotoxic effects of PEG-MTPABZ-PyC-mediated photodynamic therapy were examined using the cell counting kit-8 (CCK-8) assay and flow cytometry, while the intrinsic cytotoxicity of the photosensitizer alone was verified by the CCK-8 assay. Intracellular reactive oxygen species (ROS) generation after photodynamic therapy was detected using 2'-7'-dichlorodihydrofluorescein diacetate (DCFH-DA). RESULTS: PEG-MTPABZ-PyC alone exhibited no cytotoxicity toward MKN45 cells, indicating excellent cytocompatibility. The compound efficiently entered cells within 6 hours and localized predominantly in lysosomes. Upon light irradiation, PEG-MTPABZ-PyC-mediated photodynamic therapy induced significant cytotoxicity compared with the control group (P<0.05) and generated abundant intracellular ROS. CONCLUSIONS The novel photosensitizer PEG-MTPABZ-PyC demonstrates potent photodynamic cytotoxicity against gastric cancer cells, showing promising potential for further development in gastric cancer photodynamic therapy.
Humans ; Stomach Neoplasms/drug therapy* ; Photochemotherapy/methods* ; Photosensitizing Agents/pharmacology* ; Cell Line, Tumor ; Polyethylene Glycols/chemistry* ; Reactive Oxygen Species/metabolism* ; Mesoporphyrins/pharmacology*

Objective:To evaluate preoperative high-resolution thin-layer cervical enhanced CT used to predict the venous route of the submental flap reflux vein and its relationship with adjacent structures in order to guide the anatomical understanding and protection of submental flap in pharyngeal cancer surgery.Methods:Sixty consecutive patients with pharyngeal cancer who underwent submental flap repair surgery in our department from March 2022 to December 2024, as well as 60 patients who were accepted neck dissection suffering other cancers, were selected. Before surgery, high-resolution cervical enhanced CT scans were performed, and the position of the transverse section of the facial vein in the venous phase horizontal image gradually variation tendency was focused layer by layer. The direction and adjacent relationship of the submental flap reflux veins were determined and recorded. Combined with 60 patients with other head and neck tumors who underwent neck dissection in our department during the same period (a total of 120 cases, 240 sides), the classification and management of the draining veins of Fang′s mental flap were conducted. Type Ⅰ mainly drains into the internal jugular vein; Type Ⅱ mainly drains into the external jugular vein and Type Ⅲ mainly drains into the anterior jugular vein (often accompanied by an external jugular draining branch). The status and proportion of venous drainage were analyzed.Results:Vascular predictive coincidence rate was 98.3% (59/60) among the 60 patients with pharyngeal cancer. Only one patient was predicted to have a simple return to the external jugular vein. However, during the operation, in addition to the main return to the external jugular vein, a small portion also returned to the internal jugular vein. Submental flap reflux vessels were classified into three types based on intraoperative submental flap venous return in 60 cases of laryngopharyngeal cancer, in conjunction with the analysis of venous return patterns from 240 cervical CT scans. Type Ⅰ mainly refluxed to the internal jugular vein, accounting for 42.1%. Type Ⅱ mainly refluxed to the external jugular vein (47.9%). Type Ⅲ mainly refluxed to the anterior jugular vein (10.0%). The total detection rate of CT reading of 240 venous reflux was 98.7% (237/240). Vascular predictive coincidence rate was 97.9%(235/240).Conclusion:The detailed analysis of submental venous return vessels can accurately predict the direction of reflux veins and its surrounding areas by preoperative high-resolution enhanced CT scan. This provides reliable guidance for the anatomy and protection of the submental flap reflux veins during surgery.

Objective:To compare the survival outcomes of different subsequent treatment regimens in patients with locally advanced hypopharyngeal squamous cell carcinoma (HPSCC) who achieved partial response (PR) after neoadjuvant chemotherapy based on the gross tumor volume regression rate (GTVRR).Methods:This retrospective study included patients with locally advanced HPSCC treated at the Department of Head and Neck Surgery, Beijing Tongren Hospital, from January 2011 to December 2023. The cohort included 135 males and 3 females, aged from 35 to 77 years. All patients received 2-3 cycles of TPF regimen (paclitaxel+cisplatin+5-fluorouracil) neoadjuvant chemotherapy. Subsequent treatments included concurrent chemoradiotherapy or surgery combined with postoperative adjuvant radiotherapy. The impacts of different subsequent treatment modalities on the survivals and prognoses of patients were compared based on GTVRR thresholds of 50% and 70%. The χ 2 test was used to analyze influencing factors; survival analysis and intergroup comparisons were performed using the Kaplan-Meier method and Log-rank test; prognostic factors were assessed using univariate and multivariate Cox regression analyses. Results:The 5-year OS and PFS rates were 56.5% and 47.9%, respectively, while, the 10-year OS and PFS rates were 25.8% and 21.2%, respectively. The median OS was 75 months, and the median PFS was 48 months. The laryngeal function preservation rate for the entire cohort was 83.3%. The patients who underwent surgery combined with postoperative radiotherapy had significantly better OS and PFS outcomes than those treated with concurrent chemoradiotherapy ( P<0.05). Stratification based on GTVRR revealed that the surgery plus postoperative radiotherapy regimen was particularly effective for PR patients with a GTVRR of 30%-70%, showing significantly better OS and PFS compared to the concurrent chemoradiotherapy group ( P<0.05). Conclusion:The optimal subsequent treatment for PR-HPSCC may be surgery-based comprehensive treatment, particularly for patients with a GTVRR of 30%-70%. This study offers valuable insights for the stratified treatment of HPSCC, which could contribute to improving overall patient prognosis.

Objective:To investigate the clinical efficacy of posterior midline incision combined with anteromedial approach in the treatment of complex olecranon fracture-dislocation.Methods:A retrospective analysis was performed on 26 patients (15 males and 11 females) with olecranon fracture-dislocation who were admitted from January 2020 to January 2024, including 5 cases of anterior transolecranon fracture-dislocation (2 cases of upper ulnar-radial joint dislocation), 21 cases of posterior transolecranon fracture-dislocation (5 cases of them were accompanied by upper ulnar-radial joint dislocation). Among them, there were 13 cases of traffic accidents, 7 cases of falling from heights, and 6 cases of walking falls. The average age is 45.1±15.3 years old (21-84 years old).Results:The operation time was 151.2±41.9 minutes, average tourniquet time was 93.7±22.6 minutes, and the intraoperative blood loss was 76.2±20.2 ml. The average follow-up was 16(12, 23) months, and the VAS score decreased significantly and the MEPS score increased significantly over time. At the last follow-up, the VAS score was 2(1, 2), and the MEPS score was 86.5±10.3, with 16 cases excellent, 7 cases good, and 3 cases medium, with an excellent rate of 89%. The range of motion of flexion-extension and pronation-supination were 119.3°±13.5°and 138.6°±15.2° respectively. Complications included 16 cases of ectopic ossification, of which 4 patients with significant effects on elbow function underwent surgical release 3-6 months after surgery. 1 case of ulnar nerve injury symptoms improved after emergency ulnar nerve release, and 1 case of elbow subluxation due to inaccurate coronoid process reduction and fixation. There were no serious complications such as vascular injury, internal fixation failure, fracture nonunion, and incision infection.Conclusion:The posterior midline incision combined with anteromedial approach can effectively treat complex olecranon fracture-dislocation and meet the requirements of early postoperative elbow rehabilitation.

Objective To construct a hierarchical dynamic management intelligent system for nurses and to validate its application effectiveness,providing a reference for the development of intelligent nursing human resource management.Methods The nurse hierarchical classification and dynamic promotion management mechanism were clarified,and a hierarchical dynamic management system for nurses was established,comprising 2 modules:personal growth,and tier management and review.The system was officially implemented in a tertiary A hospital in Henan Province in March 2024.A comparison was conducted between pre-implementation(2023)and post-implementation(2024),including nurse tier applications and promotions,efficiency in tier promotion evaluations,Psychological Capital Questionnaire scores of nurses,and user satisfaction with the system.Results Following the implementation of the hierarchical dynamic management intelligent system for nurses,the application rate for tier promotion increased by 11.33％.After system implementation,the promotion pass rate and Psychological Capital Questionnaire scores of nurses were significantly higher than those before implementation,while the time required for tier review was significantly shorter.These differences were statistically significant(P＜0.05).The satisfaction survey scores for nurses applying for tier promotion and for reviewers using the system were(62.28±4.56)and(64.09±3.17)scores,respectively.Conclusion The application of the hierarchical dynamic management intelligent system for nurses helps enhance nurses'promotion pass rates and psychological capital,streamlines the nurse tier review process,and demonstrates high user satisfaction with the system.

Objective:To analyze the biological parameters of the anterior segment of age-related cataracts patients with or without zonular weakness with the new generation of sweep-source anterior optical coherence tomography (OCT) device CASIA2 to provide a basis for the diagnosis of zonular weakness.Methods:A case-control study was conducted.A total of 158 cases (186 eyes) of patients with age-related cataracts having zonular weakness as a zonular weakness group, and 80 cases (80 eyes) of cataract with age-related cataracts without zonular weakness as a normal zonule group were enroll from June 2022 to June 2023 at Beijing Tongren Hospital.All patients underwent routine preoperative ophthalmological examination including slit lamp microsopy, IOLMaster 700, ocular B-ultrasound and OCT.The anterior chamber depth (ACD), anterior chamber volume (ACV), lens thickness (LT), lens vault (LV), anterior chamber angle opening distance at 750 μm (AOD750), lens decentration and tilt, radius of curvature of the anterior lens surface (RAL), and radius of curvature of the posterior lens surface (RPL) of patients were measured with CASIA2 and compared between the two groups.The relationship between zonular weakness and anterior chamber parameters and lens parameters was evaluated by logistic regression analysis.This study followed the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Beijing Tongren Hospital (No.TRECKY2018-049), and all enrolled patients signed the informed consent form.Results:In the zonular weakness group, ACD, ACV, RAL, and AOD750 were (2.34±0.56)mm, (85.05±40.19)mm 3, 7.52 (7.13, 8.08)mm, and 0.27 (0.07, 0.30)mm, respectively, which were smaller than (3.13±0.38)mm, (127.75±38.15)mm 3, 9.28(8.51, 9.76)mm, and 0.52 (0.31, 0.65)mm in the normal zonule group, with statistically significant differences (all P<0.05).LT and LV in the zonular weakness group were (5.14±0.45)mm and (1.22±0.53)mm, respectively, which were larger than (4.27±0.52)mm and (0.32±0.30)mm in the normal zonule group, and the differences were statistically significant (both P<0.05).Magnitude of lens decentration and tilt in the zonular weakness were 0.34 (0.13, 0.45)mm and 6.44 (3.67, 7.32)°, respectively, which were significantly larger than 0.19 (0.12, 0.25)mm and 4.88 (3.85, 5.65)° in the normal zonule group (both P<0.05).No obvious pattern was found in the direction of decentration and tilt.Logistic regression analysis showed that LV and lens decentration were risk factors of zonular weakness (odds ratios [ OR]=706.170, 335.339; both P<0.05), and RAL was a protective factor of zonular weakness ( OR=0.239, P<0.05). Conclusions:Age-related cataract patients with zonular weakness often present with a decrease in ACV due to increased anterior convexity of the lens.Decreased RAL, increased lens decentration and elevated LV are risk factors for zonular weakness.