Objective:To evaluate the application of metagenomic next-generation sequencing（mNGS）in the identification of non-tuberculous mycobacteria（NTM）.Methods:A retrospective analysis was conducted on mNGS results of 358 bronchoalveolar lavage fluid（BALF）samples positive for NTM collected at the First Affiliated Hospital of Zhejiang University School of Medicine from February 2021 to January 2024. The analysis included the distribution of NTM species，the detection of mixed pathogens，and the performance of conventional mycobacterial detection methods.Results:The results showed that 362 strains of 15 NTM species were identified from 350 specimens，8 specimens were not precise to the species level. The most frequently detected species were Mycobacterium intracellulare（37.3%，135/362）， Mycobacterium abscessus（26.8%，97/362），followed by Mycobacterium avium（11.0%，40/362）， Mycobacterium kansasii（8.0%，29/362）and Mycobacterium chelonae（7.7%，28/362）. Single NTM species were detected in 339 specimens，while two or three NTM species were simultaneously detected in 11 specimens（3.1%，11/358）. Non-NTM microorganisms co-infected were detected in 53.4%（191/358）of NTM-positive BALF samples，including common pathogens such as Pseudomonas aeruginosa， Staphylococcus aureus，and Aspergillus fumigatus；and difficult-to-identify pathogens such as Legionella pneumophila and Talaromyces marneffei. In NTM-positive patients detected by mNGS，the results supported the diagnosis of NTM infection in 298 cases（298/358，83.2%）and 105 cases（105/358，29.3%）initiated anti-NTM treatment accordingly；while in 60 cases（60/358，16.8%）the positive results were considered as colonization or unrelated to clinical infection. For samples tested with acid-fast staining，mycobacterial liquid culture，and DNA microarray，the positivity rates for NTM were 31.5%（73/232），48.7%（57/117），and 43.0%（46/107），respectively. Conclusions:mNGS demonstrates advantages in identification of NTM. However，the test may detect multiple microorganisms，in that case，the interpretation with clinical and radiological results is requried to determine the main pathogens.

Pyruvate kinase M2(PKM2)is closely linked to a variety of neurological disorders,involved in the onset and progression of a wide range of diseases,including Alzheimer's disease,Parkinson's disease,multiple sclerosis and traumatic brain injury through such mechanisms as aerobic glycolysis,oxidative stress,inflammation and apoptosis.This review is intended to provide an overview of the biological characteristics of PKM2 and its role in the pathophysiological mechanisms of neurological disorders.A better understanding of the correlations between PKM2 and the development of neurological diseases can offer new insights into and references for the clinical diagnosis and treatment of these conditions.

Objective To investigate the nutritional quality characteristics of vitamin D3-fortified yogurt and explore its improving effect on vitamin D metabolism in the body under simulated weightlessness,thereby providing a theoretical basis for the development of functional foods.Methods Using reconstituted milk as the matrix and Vitamin D3(VD3)microcapsule powder as the fortifier,VD3-fortified yogurt was prepared.A systematic study was conducted to investigate the effects of different gradients(1.25 μg/100 mL,2.50 μg/100 mL,3.75 μg/100 mL,5.00 μg/100 mL,6.25 μg/100 mL)of VD3 microcapsule addition on its quality characteristics(titratable acidity,solid content,water-holding capacity,syneresis).In vivo assessments were conducted using a Sprague-Dawley(SD)rat tail-suspension model to simulate weightlessness.Levels in serum 25(OH)D3,1,25-(OH)2D3,calcium(Ca),and phosphorus(P)were detected using the enzyme-linked immunosorbent assay(ELISA)to evaluate its metabolic capacity.Results During fermentation(3 h),titratable acidity of VD?-fortified yogurt initially increased,then decreased,and eventually stabilized with rising microcapsule dosage,while total solid content remained consistent.WHC exhibited an initial increase followed by a decline,whereas syneresis showed an inverse trend.At an optimal dosage of 3.75 μg/100 mL,the yogurt displayed a dense and uniform network structure,characterized by non-Newtonian fluid behavior with shear-thinning properties.This formulation demonstrated robust structural stability under high-frequency mechanical stress,alongside desirable textural,flavor,and sensory attributes.Animal experiments revealed that the serum concentrations of 25(OH)D3,1,25-(OH)2D3,calcium,and phosphorus in the vitamin D?-fortified yogurt intervention group were significantly higher than those in the tail-suspended control group(P<0.05).Conclusion VD? microencapsulation technology effectively preserves and enhances the nutritional quality characteristics of yogurt and mitigates vitamin D metabolic dysregulation under simulated weightlessness.

A 49-year-old female patient with recurrent cervical cancer was treated with injectable paclitaxel(albumin-bound)(400 mg,ivd,d1)+injectable cisplatin 100 mg,intraperitoneal instillation,d1+candonilimab injection(375 mg,ivd,d7).During the 2nd cycle of treatment,when the intravenous infusion of candonilimab was about 8 minutes,the patient suddenly had symptoms such as flushing,shortness of breath,cardiac discomfort,blurred consciousness with profuse sweating,etc.The candonilimab was immediately suspended,and after resuscitation,the patient's vital signs stabilised,and the treatment of the primary disease with candonilimab was terminated.According to the principle of correlation evaluation in Adverse Drug Reaction Reporting and Monitoring Workbook,the correlation between anaphylaxis and candonilimab was evaluated to be"very likely",and the common types of adverse reactions to candonilimab,the mechanism of anaphylaxis and pretreatment measures were also reviewed,with a view to providing referable opinions on the whole course of management of candonilimab and promoting the rational use of the drug in clinical safety.

Objective:To evaluate the application of metagenomic next-generation sequencing（mNGS）in the identification of non-tuberculous mycobacteria（NTM）.Methods:A retrospective analysis was conducted on mNGS results of 358 bronchoalveolar lavage fluid（BALF）samples positive for NTM collected at the First Affiliated Hospital of Zhejiang University School of Medicine from February 2021 to January 2024. The analysis included the distribution of NTM species，the detection of mixed pathogens，and the performance of conventional mycobacterial detection methods.Results:The results showed that 362 strains of 15 NTM species were identified from 350 specimens，8 specimens were not precise to the species level. The most frequently detected species were Mycobacterium intracellulare（37.3%，135/362）， Mycobacterium abscessus（26.8%，97/362），followed by Mycobacterium avium（11.0%，40/362）， Mycobacterium kansasii（8.0%，29/362）and Mycobacterium chelonae（7.7%，28/362）. Single NTM species were detected in 339 specimens，while two or three NTM species were simultaneously detected in 11 specimens（3.1%，11/358）. Non-NTM microorganisms co-infected were detected in 53.4%（191/358）of NTM-positive BALF samples，including common pathogens such as Pseudomonas aeruginosa， Staphylococcus aureus，and Aspergillus fumigatus；and difficult-to-identify pathogens such as Legionella pneumophila and Talaromyces marneffei. In NTM-positive patients detected by mNGS，the results supported the diagnosis of NTM infection in 298 cases（298/358，83.2%）and 105 cases（105/358，29.3%）initiated anti-NTM treatment accordingly；while in 60 cases（60/358，16.8%）the positive results were considered as colonization or unrelated to clinical infection. For samples tested with acid-fast staining，mycobacterial liquid culture，and DNA microarray，the positivity rates for NTM were 31.5%（73/232），48.7%（57/117），and 43.0%（46/107），respectively. Conclusions:mNGS demonstrates advantages in identification of NTM. However，the test may detect multiple microorganisms，in that case，the interpretation with clinical and radiological results is requried to determine the main pathogens.

A 49-year-old female patient with recurrent cervical cancer was treated with injectable paclitaxel(albumin-bound)(400 mg,ivd,d1)+injectable cisplatin 100 mg,intraperitoneal instillation,d1+candonilimab injection(375 mg,ivd,d7).During the 2nd cycle of treatment,when the intravenous infusion of candonilimab was about 8 minutes,the patient suddenly had symptoms such as flushing,shortness of breath,cardiac discomfort,blurred consciousness with profuse sweating,etc.The candonilimab was immediately suspended,and after resuscitation,the patient's vital signs stabilised,and the treatment of the primary disease with candonilimab was terminated.According to the principle of correlation evaluation in Adverse Drug Reaction Reporting and Monitoring Workbook,the correlation between anaphylaxis and candonilimab was evaluated to be"very likely",and the common types of adverse reactions to candonilimab,the mechanism of anaphylaxis and pretreatment measures were also reviewed,with a view to providing referable opinions on the whole course of management of candonilimab and promoting the rational use of the drug in clinical safety.

Objective:To investigate the effect of different timing of arterial -venous extracorporeal membrane oxygenation (VA-ECMO) on the prognosis of patients with acute myocardial infarction complicated with cardiogenic shock (AMICS).Methods:This study was a prospective cohort study. AMICS patients received VA-ECMO support primary percutaneous coronary intervention in Henan Provincial People's Hospital from May 2017 to July 2023 were divided into early VA-ECMO group and late VA-ECMO group. 64 AMICS patients who met the indications for VA-ECMO implantation, but did not revive VA-ECMO were included as control group. Demographic characteristics, coronary interventional (PCI) information and complications after VA-ECMO implantation were collected. The primary end points was 1-year survival, minor end point were in-hospital and perioperative death. Multivariate Logistic and Cox regression models were used to evaluate the effect of timing of VA-ECMO on prognosis of AMICS patients. Kaplan-Meier survival curve was used to analyze the 1-year survival outcome of the 3 groups.Results:A total of 143 AMICS patients were included, and materials of 136 patients entered in the final analysis, including 42 in the early VA-ECMO group, 34 in the late VA-ECMO group, and 60 in the non-VA-ECMO group. Compared with the late VA-ECMO group, the early VA-ECMO group had a higher ratio of PPCI after VA-ECMO, a longer D-to-B time, a shorter VA-ECMO support time, a higher success rate of VA-ECMO withdrawal, and a lower complication rate (all P<0.05). Compared with the early VA-ECMO group, the perioperative, in-hospital and 1-year mortality were significantly higher in Non-ECMO support (all P<0.05). There was no difference in perioperative and in-hospital mortality between the early VA-ECMO group and the late VA-ECMO group, but the 1-year mortality in the late VA-ECMO group was significantly higher ( P<0.05). Perioperative, in-hospital and 1-year mortality rates were lower in the late VA-ECMO group than in the no-VA-ECMO group, but the differences were not statistically significant. Multivariate Logistic and Cox regression analysis showed that after adjusting interference factors, early VA-ECMO was still a protective factor for in-hospital ( OR=0.244, P=0.015) and one year ( HR=0.308, P=0.001)mortality. Kaplan-Merier survival curve showed that compared with the late VA-ECMO group and the group without VA-ECMO, the early VA-ECMO group had the highest 1-year survival rate. Conclusion:Patients with AMICS may benefit more from early VA-ECMO than from late VA-ECMO support for PPCI.

Objective:To explore the feasibility of circulating tumor cell (CTC) dual-antibody enrichment and dual-antibody detection for epithelial-mesenchymal phenotypes in patients with hepatocellular carcinoma (HCC), and investigate the clinical diagnostic value of CTC typing in evaluating postoperative recurrence and prognosis of HCC.Methods:Of 89 HCC patients who underwent surgical treatment in Zhejiang Jinhua Guangfu Tumor Hospital from March 2020 to January 2024 were enrolled into this study, including 73 males and 16 females, aged (64.4±9.5) years old. The peripheral blood samples of patients were collected before operation. Epithelial CTC, mesenchymal CTC and hybrid epithelial-mesenchymal CTC in blood samples of patients with HCC were enriched and detected by EpCAM/CSV double capture antibodies and PanCK/CSV double detection antibodies. Kaplan-Meier analysis was employed to assess recurrence-free survival (RFS) and overall survival (OS) rates. Univariate and multivariate Cox regression were used to analyze the effects of different types of CTC on postoperative RFS and OS.Results:The detection rates of total CTC, epithelial CTC, mesenchymal CTC and hybrid epithelial-mesenchymal CTC were 92.1% (82/89), 64.0% (57/89), 62.9% (56/89) and 55.1% (49/89), respectively. Multivariate Cox regression analysis showed that HCC patients with more mesenchymal CTC ( HR=2.408, 95% CI: 1.580-3.668) and hybrid epithelial-mesenchymal CTC ( HR=1.840, 95% CI: 1.004-3.371) in peripheral blood had higher postoperative recurrence risk (both P<0.05). Univariate Cox regression showed patients with more total CTC ( HR=1.426, 95% CI: 1.040-1.954, P=0.028) was associated with survival. Conclusion:The technique of epithelial-mesenchymal phenotypes analysis of circulating tumor cells based on dual-antibody capture and dual-antibody detection is feasible. The number of mesenchymal CTC and hybrid epithelial-mesenchymal CTC before operation is the influencing factor of postoperative recurrence in patients with hepatocellular carcinoma.

Objective:To explore the changes of gait flexibility and stability in patients with Parkinson's disease (PD) 2 years after deep brain stimulation (DBS).Methods:Twenty PD patients accepted DBS in Department of Neurosurgery, Tianjin Huanhu Hospital from October 2019 to November 2021 were enrolled. Motor symptoms were evaluated by Movement Disorder Society-unified Parkinson's disease rating scale III (MDS-UPDRS-III) at preoperative medication-off state, postoperative medication-off state, and postoperative medication-on state. Mini-mental state examination (MMSE) and Montreal cognitive assessment (MoCA) were used to assess the cognition and 39-item Parkinson's disease questionnaire (PDQ-39) was used to evaluate the quality of life at preoperative medication-on state and postoperative medication-on state. A three-dimensional gait analyzer was used to record the gait parameters during Instrumented Stand and Walk test (ISAW) at preoperative medication-off state, postoperative medication-off state, and postoperative medication-on state. Differences in motor symptom scores, cognitive scores, quality of life scores, as well as changes in gait flexibility and stability were compared before and after DBS.Results:(1) The MDS-UPDRS-Ⅲ scores at preoperative medication-off state, postoperative medication-off state, and postoperative medication-on state ([45.30±12.57], [24.95±10.74], [15.80±7.19]) were decreased successively, with significant differences ( P<0.05).(2) Compared with those before surgery, PD patients had significantly lower levodopa equivalent daily dose (LEDD), total scores of PDQ-39, and scores of question 9 "degree of concern about falling" in PDQ-39 at 2 years after DBS ([711.84±343.99] mg/d vs. [549.30±301.08] mg/d, 47.00[30.00, 64.00] vs. 13.50[7.75, 27.00], 2.00[0.00, 3.00] vs. 0.00[0.00, 1.75], P<0.05). (3) Compared with that at preoperative medication-off state, the arm swing velocity at postoperative medication-on state statistically increased in PD patients ( P<0.05); compared with those at preoperative medication-off state, the arm swing range and turning speed at postoperative medication-off and medication-on states significantly increased in PD patients ( P<0.05); compared with that at preoperative medication-off state, the turning duration at postoperative medication-off state statistically decreased in PD patients ( P<0.05). Compared with that at preoperative medication-off state, the range of motion of the trunk in the horizontal plane at postoperative medication-off and medication-on states increased significantly in PD patients ( P<0.05); compared with that at preoperative medication-off state, the range of motion in the sagittal plane of the lumbar, coronal plane of the trunk, and sagittal plane of the trunk all increased significantly in PD patients at postoperative medication-on state ( P<0.05); the mean velocity and root mean square acceleration at postoperative medication-on state increased significantly in PD patients compared with that at preoperative medication-off state ( P<0.05); conversely, the swing frequency at postoperative medication-off state decreased significantly in PD patients compared with that at preoperative medication-off state ( P<0.05). Conclusion:Two years after DBS, PD patients exhibit obviously improved gait, with enhanced flexibility, and dynamic and static stability.

Objective:To investigate the incidence and potential risk factors associated with postopera-tive spinal epidural hematoma(SEH)following anterior cervical spine surgery(ACSS).Methods:A retrospective analysis was conducted on the clinical data of patients who underwent ACSS for cervical spondylosis at Peking University Third Hospital between March 2013 and February 2022.Patients who developed postoperative SEH were categorized as the SEH group,while those in the cohort without SEH were randomly selected as the non-SEH group by individually matching with the same operator,same gender,same surgery year,and similar age(±5 years)at a ratio of 4:1.The general condition,pre-operative comorbidities,anticoagulant or antiplatelet therapy,preoperative coagulation and platelet counts,American society of Anesthesiologists physical status classification,cervical spondylosis classifi-cation,preoperative modified Japanese Orthopaedic Society score and cervical disability index score,sur-gical modality,surgical segment levels,ossification of the posterior longitudinal ligament among the surgi-cal level,surgery duration,estimated blood loss,postoperative drainage volume,preoperative mean arte-rial pressure,mean arterial pressure during postoperative awakening periods,hospital stay and hospitali-zation cost were compared between the two groups.A bivariate Logistic regression model was applied to screen out the independent risk factors and calculate the odds ratios of indicators associated with SEH.Receiver operating characteristic curve and area under the curve(AUC)were used to describe the dis-crimination ability of the indicators.Results:A total of 85 patients were enrolled in the study,including 17 patients in the SEH group and 68 patients in the non-SEH group.Seventeen patients with SEH under-went hematoma evacuation,and all of them were successfully treated and discharged from the hospital.Corpectomy(OR=7.247;95％CI:1.962-26.766;P=0.003)and the highest mean arterial pressure during awakening(OR=1.056;95％CI:1.002-1.113;P=0.043)were independent risk factors for SEH.The AUC values were 0.713(95％CI:0.578-0.848)and 0.665(95％CI:0.51-0.82)re-spectively.The patients with SEH had longer hospital stays(P＜0.001)and greater hospitalization costs(P=0.035).Conclusion:Corpectomy and elevated maximum mean arterial pressure during awakening are independent risk factors for the development of postoperative SEH following ACSS.High-risk patients should be closely monitored during the perioperative period.