ObjectiveTo prepare a recombinant PETase with a dual-catalytic-triad and to evaluate its efficiency in the biodegradation of polyethylene terephthalate （PET）. MethodsBased on the crystal structure of wild-type PETase， point mutations （T88H/L117D） were introduced via site-directed mutagenesis. The recombinant protein was prepared using prokaryotic expression and chromatography purification techniques. The enzymatic hydrolysis of the mutant PETase was assessed by relatively quantifying the products mono （2-hydroxyethyl） terephthalate （MHET） and terephthalic acid （TPA）. ResultsBoth wild-type and mutant PETases accumulated as inclusion bodies， accounting for approximately 20% of the total bacterial protein. After solubilization in urea， the proteins were eluted at 300 mmol/L imidazole during affinity chromatography purification， with concentrations of 1.824 and 1.833 mg/mL and purities of 83.11% and 84.32%， respectively. Subsequent anion-exchange chromatography yielded highly pure enzymes in the 200 mmol/L NaCl fraction： 2.776 mg/mL （96.86% purity） for the wild type and 1.967 mg/mL （95.13% purity） for the mutant. Following refolding， the final concentrations were 0.484 mg/mL for the wild type and 0.991 mg/mL for the mutant. Hydrolysis assays revealed that the mutant released MHET and TPA at （237.67±17.00）% and （197.33±12.01）% of the wild-type levels， respectively. ConclusionThe T88H/L117D dual-catalytic-triad PETase is successfully prepared and it significantly enhanced PET-degrading activity， thus， it′s a promising biocatalyst for PET bioremediation.

Oral squamous cell carcinoma(OSCC)is a common malignant tumor.Interleukin-8(IL-8)is an important biomarker of OSCC,and its level can reflect the occurrence and development of OSCC.It is of great significance to detect IL-8 rapidly and sensitively for the purpose of early diagnosis of OSCC.In this study,gold nanoparticles(AuNPs)with uniform particle size were synthesized by reduction of chloroauric acid with trisodium citrate,and the probe(AuNPs@mAb1)was prepared by coupling AuNPs with the murine anti-IL-8 monoclonal antibody mAb1.Cy5-NHS and murine anti-IL-8 monoclonal antibody mAb2 complex(Cy5-mAb2)and sheep anti-mouse IgG antibody were sprayed on nitrocellulose membrane to form test line(T line)and control line(C line)respectively,and reverse fluorescence-enhanced test strips were thus constructed,based on which an immunochromatographic method was established for highly sensitive detection of IL-8 in saliva samples.The experimental results showed that the test strip had good stability,high specificity and high sensitivity.The linear range for fluorescence detection of IL-8 was 0.01-100 ng/mL,and the limit of detection(3σ)was 7.93 pg/mL.The linear range for visualization detection was 6-100 ng/mL,with limit of detection(3σ)of 0.85 ng/mL.The fabricated test trip had good preparation reproducibility,with inter-and intra-batch assay precision of less than 5.5%.The test strip was used for detection of IL-8 in healthy human saliva samples,with spiked recoveries of 93.7% -102.4%,and relative standard deviations of 2.1% -4.3%.The fabricated test strip could be used for early screening of OSCC.

Approximately 30%-40% of epilepsy patients do not respond well to adequate anti-seizure medications (ASMs), a condition known as pharmacoresistant epilepsy. The management of pharmacoresistant epilepsy remains an intractable issue in the clinic. Its early prediction is important for prevention and diagnosis. However, it still lacks effective predictors and approaches. Here, a classical model of pharmacoresistant temporal lobe epilepsy (TLE) was established to screen pharmacoresistant and pharmaco-responsive individuals by applying phenytoin to amygdaloid-kindled rats. Ictal electroencephalograms (EEGs) recorded before phenytoin treatment were analyzed. Based on ictal EEGs from pharmacoresistant and pharmaco-responsive rats, a convolutional neural network predictive model was constructed to predict pharmacoresistance, and achieved 78% prediction accuracy. We further found the ictal EEGs from pharmacoresistant rats have a lower gamma-band power, which was verified in seizure EEGs from pharmacoresistant TLE patients. Prospectively, therapies targeting the subiculum in those predicted as "pharmacoresistant" individual rats significantly reduced the subsequent occurrence of pharmacoresistance. These results demonstrate a new methodology to predict whether TLE individuals become resistant to ASMs in a classic pharmacoresistant TLE model. This may be of translational importance for the precise management of pharmacoresistant TLE.
Epilepsy, Temporal Lobe/diagnosis* ; Animals ; Drug Resistant Epilepsy/drug therapy* ; Electroencephalography/methods* ; Rats ; Anticonvulsants/pharmacology* ; Neural Networks, Computer ; Male ; Humans ; Phenytoin/pharmacology* ; Adult ; Disease Models, Animal ; Female ; Rats, Sprague-Dawley ; Young Adult ; Convolutional Neural Networks

Apical microsurgery is accurate and minimally invasive, produces few complications, and has a success rate of more than 90%. However, due to the lack of awareness and understanding of apical microsurgery by dental general practitioners and even endodontists, many clinical problems remain to be overcome. The consensus has gathered well-known domestic experts to hold a series of special discussions and reached the consensus. This document specifies the indications, contraindications, preoperative preparations, operational procedures, complication prevention measures, and efficacy evaluation of apical microsurgery and is applicable to dentists who perform apical microsurgery after systematic training.
Microsurgery/standards* ; Humans ; Apicoectomy ; Contraindications, Procedure ; Tooth Apex/diagnostic imaging* ; Postoperative Complications/prevention & control* ; Consensus ; Treatment Outcome

Intentional tooth replantation (ITR) is an advanced treatment modality and the procedure of last resort for preserving teeth with inaccessible endodontic or resorptive lesions. ITR is defined as the deliberate extraction of a tooth; evaluation of the root surface, endodontic manipulation, and repair; and placement of the tooth back into its original socket. Case reports, case series, cohort studies, and randomized controlled trials have demonstrated the efficacy of ITR in the retention of natural teeth that are untreatable or difficult to manage with root canal treatment or endodontic microsurgery. However, variations in clinical protocols for ITR exist due to the empirical nature of the original protocols and rapid advancements in the field of oral biology and dental materials. This heterogeneity in protocols may cause confusion among dental practitioners; therefore, guidelines and considerations for ITR should be explicated. This expert consensus discusses the biological foundation of ITR, the available clinical protocols and current status of ITR in treating teeth with refractory apical periodontitis or anatomical aberration, and the main complications of this treatment, aiming to refine the clinical management of ITR in accordance with the progress of basic research and clinical studies; the findings suggest that ITR may become a more consistent evidence-based option in dental treatment.
Humans ; Tooth Replantation/methods* ; Consensus ; Periapical Periodontitis/surgery*

ObjectiveTo investigate the association of SH2B1 rs7359397 （C>T） polymorphism with the progression to hepatic fibrosis in the elderly patients with metabolic dysfunction-associated steatotic liver disease （MASLD） in Beijing， China， and to provide an important genetic basis for the precise subtyping， prognostic evaluation， and individualized treatment of elderly MASLD patients in China. MethodsA total of 505 elderly patients （aged ≥65 years） who participated in regular physical examination in Mentougou Kuangshan Hospital of Beijing Jingmei Group General Hospital from November 2020 to September 2021 and were diagnosed with MASLD by abdominal ultrasound were enrolled as MASLD group， and 381 elderly population who underwent physical examination in the same community hospital during the same period of time and were not found to have MASLD by abdominal ultrasound were enrolled as control group. FibroScan was used to measure liver fat content and determine fibrosis stage. The 96-well microfluidic chip technique was used to identify SH2B1 rs7359397 polymorphism. The independent-samples t test was used for comparison of normally distributed continuous data between the two groups， and the chi-square test or the adjusted chi-square test was used for comparison of categorical data between the two groups. Univariate and multivariate Logistic regression analyses were used to identify the independent predictive factors for MASLD and its comorbidities. ResultsCompared with the control group， the MASLD group had a significantly younger age and significantly higher levels of waist circumference， hip circumference， waist-hip ratio， body mass index （BMI）， alanine aminotransferase， aspartate aminotransferase， triglyceride， platelet count， and fibrosis-4 （FIB-4） index， as well as a significantly lower level of high-density lipoprotein cholesterol （all P<0.05）. Among the 381 patients in the control group， 264 （69.29%） had genotype CC and 117 （30.71%） had genotype CT+TT， while among the 505 patients in the MASLD group， 317 （62.77%） had genotype CC and 188 （37.23%） had genotype CT+TT， suggesting that the MASLD group had a significantly higher proportion of patients with genotype CT+TT compared with the control group （χ²=4.09， P=0.043）. In the MASLD group， compared with the genotype CC group， the genotype CT+TT group had a significantly lower proportion of patients with FIB-4 ≥2 or atherosclerotic cardiovascular diseases （P<0.05）. The multivariate Logistic regression analysis showed that after adjustment for age， sex， and BMI， carrying T allele was a protective factor against progressive hepatic fibrosis （odds ratio ［OR］=0.481， 95% confidence interval ［CI］： 0.249 — 0.929， P=0.029）. In the subgroups of comorbidities with hypertension， metabolic syndrome， and obesity， genotype CT+TT was associated with a significant reduction in the risk of progressive hepatic fibrosis （hypertension： OR=0.27， 95%CI：0.09 — 0.77， P=0.014； metabolic syndrome： OR=0.30， 95%CI： 0.11 — 0.79， P=0.015； obesity： OR=0.11， 95%CI： 0.03 — 0.48， P=0.003）. After adjustment for age， sex， and BMI， in the patients with MASLD， the patients with genotype CT+TT had a significant reduction in the prevalence rate of atherosclerotic cardiovascular diseases compared with those with genotype CC （OR=0.506， 95%CI：0.336 — 0.761， P=0.001）. ConclusionSH2B1 rs7359397 （C>T） polymorphism is associated with the reduction in the risk of hepatic fibrosis and atherosclerotic cardiovascular diseases in MASLD patients.

Objective To investigate the pharmacological mechanism through which total flavonoids extracted from Xiaobuxin-Tang(XBXT-2)affects neural network activities in the frontal cortex by focusing on the effects of XBXT-2 on the cortical field potentials in the frontal association cortex(FrA)in mice.Methods Cortical electrodes were implanted into the skull of C57BL/6J mice targeting the FrA.After a 7-day recovery period,the mice were administered XBXT-2 intragastrically at a dose of 100 mg/kg,and 1 hour later,local field potential(LFP)in the FrA were recorded for 30 minutes.Spectral analysis of the data was performed using Neuro Explorer software.Changes in the power spectral density of α,β,θ,γ,and δ frequency bands before and after drug administration were analyzed using GraphPad Prism 10.3.Phase-amplitude coupling of θ and γ oscillations was analyzed using Matlab 2021 software.Results It was found that the oral administration of XBXT-2 significantly suppressed high-frequency γ oscillations while simultaneously enhancing θ,β,α,and δ oscillations in FrA of mice compared to the control.Furthermore,XBXT-2 treatment markedly strengthened the phase-amplitude coupling between θ and γ oscillations.Conclusion XBXT-2 possibly affects emotional and cognitive functions by modulating neural network activity in FrA and enhancing θ-γ phase-amplitude coupling in mice.

Objective To establish a stable,reliable,and clinically relevant porcine model of endotoxin-induced acute respiratory distress syndrome(ARDS).Methods Ten 8-month-old male Bama minipigs were deeply sedated,followed by invasive mechanical ventilation and electrocardiographic monitoring.Lipopolysaccharide(LPS)was intravenously pumped at 600 μg/(kg·h)for 3 hours,then maintained at 15 μg/(kg·h)thereafter.Dynamic monitoring was performed at five time points after LPS injection(LPS 0,1,3,5,and 8 h),including arterial blood gas analysis and chest computed tomography(CT)scans.Pathological examination of lung tissues obtained via bronchoscopic biopsy(HE staining and transmission electron microscopy)was conducted.These indicators were comprehensively used to evaluate the success of the animal model.Results At 5 hours after LPS administration,8 minipigs developed symptoms such as skin cyanosis,elevated body temperature,and respiratory distress.The oxygenation index decreased to<300 mmHg.Chest CT scans showed diffuse pulmonary infiltrates.Histopathology revealed alveolar edema and hyaline membrane formation.Transmission electron microscopy demonstrated disruption of pulmonary blood-air barrier,depletion of lamellar bodies in type Ⅱ pneumocytes,inflammatory cell infiltration,and exudation of plasma proteins and fibrin.Compared with LPS 0 h,at LPS 8 h,the oxygenation index and arterial blood pH were significantly decreased(P<0.001),while blood lactic acid and serum potassium were significantly increased(P<0.05);serum calcium and base excess were significantly decreased(P<0.05),and the lung injury score based on HE-stained lung sections was significantly increased(P<0.01).Conclusion The porcine ARDS model established by continuous LPS injection can dynamically simulate the pathophysiological characteristics and typical pathological manifestations of clinical septic ARDS,making it an effective tool to study the pathogenesis,prevention,and treatment strategies of septic ARDS.

Objective:To investigate the predictive value of interventricular septal thickness(IVST),the ratio of mitral ear-ly diastolic peak flow velocity to late diastolic peak flow velocity(E/A)combined with aortic valve flow velocity(AV)for hypertensive heart disease(HHD)and its association with blood pressure grade.Methods:A total of 202 patients with hy-pertension suspected of HHD admitted to Qinhuangdao Hospital of Dongfang Hospital,Beijing University of Chinese Medi-cine between May 2020 and May 2023 were enrolled and divided into HHD group(n=102)and non-HHD group(n=100)according to color Doppler energy imaging and blood pressure diagnosis.Blood pressure,IVST,E/A and AV were measured in two groups.The predictive value of IVST,E/A and AV for HHD were evaluated by receiver operating charac-teristic(ROC)curve.According to hypertension grading,patients in HHD group was divided into grade 1～2 group(n=59)and grade 3 group(n=41).The association of IVST,E/A,AV with blood pressure grade in HHD patients were ana-lyzed by Spearman correlation analysis.Results:Compared with patients in non-HHD group,those in HHD group had significant higher IVST,and significant lower E/A and AV(P＜0.001 all);Compared with patients in grade 1～2 group,those in grade 3 group had significant higher IVST,and significant lower E/A and AV(P＜0.001 all).ROC curve showed that the AUC of IVST,E/A and AV alone and their combination in HHD diagnosis was 0.798(95％CI 0.736～0.851),0.682(95％CI 0.613～0.746),0.810(95％CI 0.748～0.862)and 0.927(95％CI 0.881～0.959)respectively,and the diagnostic value of combined detection was significantly higher than those of single detections(Z=4.431,5.669,3.948,P=0.001 all).Spearman correlation analysis showed that E/A and AV were significant negatively correlated with blood pressure grade(r=-0.725,-0.781),while I VST was significant positively correlated with blood pressure grade(r=0.714),and combined detection possessed a stronger correlation with it in HHD patients(r=0.875,P＜0.001 all).Con-clusion:The combined detection of I VST,E/A and AV demonstrates a good predictive value for HHD and exhibits a sig-nificant correlation with blood pressure grade in HHD patients.

Objective:To investigate the incidence of shoulder work-related musculoskeletal disorders (WMSDs) among occupational population in China, and to explore their intrinsic association with personal and work-related factors.Methods:In April 2024, 73497 valid questionnaires of the Chinese version of the Musculoskeletal Disorders Electronic Questionnaire were retrospectively analyzed from June 2018 to December 2023 in 22 provinces and 29 key industries in China, and the general information, occurrence of WMSDs and related risk factors of key occupational populations in different regions in China were collected. By using Chi-square test and confirmatory factor analysis, the relationship between shoulder fatigue and pain in key occupational groups and individual factors, work type, work posture and work organization was discussed, and the internal relationship was analyzed based on structural equation model.Results:Higher incidence of shoulder fatigue and pain were associated with female, lack of physical exercise, uncomfortable working posture and neck leaning forward ( P<0.05). Structural equation model analysis showed that work type, work posture and work organization were strongly correlated ( r=0.58, 0.55). Work organization and work type were strongly correlated with shoulder fatigue ( r=0.65) and moderately correlated with shoulder fatigue ( r=0.21). Shoulder fatigue was moderately associated with shoulder pain ( r=0.40). Individual factors, work type, work posture and shoulder fatigue could directly affect shoulder pain ( OR=0.07, -0.09, 0.17 and 0.40), and work type and work posture could also indirectly affect shoulder pain through shoulder fatigue ( OR=0.08, 0.03). Work organization only indirectly affected shoulder pain through shoulder fatigue ( OR=0.26) . Conclusion:The main influencing factor of shoulder pain is shoulder fatigue, followed by work posture and individual factors. Structural equation model can better reflect the complex relationship between work type, work posture and work organization and shoulder WMSDs. Improving work posture and work organization may be an effective way to control the influence of shoulder fatigue on shoulder pain.