Objective This study aims to achieve high-yield functional expression of the human voltage-gated potassium channel K_V3.1 using an Escherichia coli cell-free protein synthesis system, thereby providing a novel synthetic approach for drug screening, structural analysis and functional characterization of K_V3.1. Methods K_V3.1 was expressed in an Escherichia coli cell-free protein synthesis system for 10 hours in the presence of peptide surfactant A₆K. The secondary structure of K_V3.1 was analyzed by circular dichroism spectroscopy. The potassium channel activity of the recombinant protein liposome K_V3.1-A₆K was investigated using fluorescent dyes Oxonol VI as indicators, which are capable of reflecting alterations in membrane potential. Results Soluble K_V3.1 protein was successfully synthesized, achieving a purified yield of up to 1.2 mg/mL via an Escherichia coli cell-free protein synthesis system. Circular dichroism spectroscopy revealed that K_V3.1 exhibited characteristic α-helical secondary structures. Membrane potential fluorescence assays demonstrated that the K_V3.1-A₆K proteoliposomes, which were reconstructed with surfactant peptide A₆K, exhibited remarkable potassium ion permeability. Conclusion This study successfully achieved high-yield expression of human K_V3.1 with activity using an Escherichia coli-based cell-free protein synthesis system. This innovative method not only significantly enhances the expression yield of K_V3.1, but also maintains its functional activity, thereby establishing a novel and efficient synthetic platform for drug screening and advancing our understanding of structure-function relationships in K_V3.1 research.
Humans ; Escherichia coli/metabolism* ; Shaw Potassium Channels/biosynthesis* ; Cell-Free System ; Circular Dichroism ; Protein Biosynthesis ; Recombinant Proteins/metabolism* ; Membrane Potentials ; Shab Potassium Channels

As activated hepatic stellate cells (aHSCs) play a central role in fibrogenesis, they have become key target cells for anti-fibrotic treatment. Nevertheless, the therapeutic efficiency is constrained by the exosomes they secrete, which are linked to energy metabolism and continuously stimulate the activation of neighboring quiescent hepatic stellate cells (qHSCs). Herein, an intercellular communication interference strategy is designed utilizing paeoniflorin (PF) loaded and hyaluronic acid (HA) coated copper-doped ZIF-8 (PF@HA-Cu/ZIF-8, PF@HCZ) to reduce energy-related exosome secretion from aHSCs, thus preserving neighboring qHSCs in a quiescent state. Simultaneously, the released copper and zinc ions disrupt key enzymes involved in glycolysis to reduce bioenergy synthesis in aHSCs, thereby promoting the reversion of aHSCs to a quiescent state and further decreasing exosome secretion. Therefore, PF@HCZ can effectively sustain both aHSCs and qHSCs in a metabolically dormant state to ultimately alleviate liver fibrosis. The study provides an enlightening strategy for interrupting exosome-mediated intercellular communication and remodeling the energy metabolic status of HSCs with boosted antifibrogenic activity.

Early correction of childhood malocclusion is timely managing morphological, structural, and functional abnormalities at different dentomaxillofacial developmental stages. The selection of appropriate imaging examination and comprehensive radiological diagnosis and analysis play an important role in early correction of childhood malocclusion. This expert consensus is a collaborative effort by multidisciplinary experts in dentistry across the nation based on the current clinical evidence, aiming to provide general guidance on appropriate imaging examination selection, comprehensive and accurate imaging assessment for early orthodontic treatment patients.
Humans ; Malocclusion/diagnostic imaging* ; Child ; Consensus

OBJECTIVE: To characterize a glycosyltransferase (RpUGT1) from Rheum palmatum and investigate its specificity toward flavonoid compounds. METHODS: The RpUGT1 was expressed in Escherichia coli and screened for catalytic activity against a range of flavonoid substrates using a high-throughput HPLC assay method. Mass spectrometry (MS) and nuclear magnetic resonance (NMR) were used to determine the structure of the product. Homology modeling, molecular docking analyses and site-directed mutagenesis studies were conducted to identify key residues responsible for its function. RESULTS: The recombinant RpUGT1 protein exhibited catalytic activity towards various flavonoids. Notably, RpUGT1 catalyzed the glycosylation of isorhamnetin to form 3-O-glucoside and kaempferol to form 7-O-glucoside, utilizing uridine diphosphate (UDP) glucose as the sugar donor. The homology modeling and molecular docking analyses identified key residues responsible for its activity. Subsequent site-directed mutagenesis studies highlighted the crucial role of K307 in catalysis. CONCLUSION These discoveries offer valuable perspectives on the role of the UGT family and establish a groundwork for forthcoming research on the synthesis of flavonoids in plants.

OBJECTIVE To investigate the predictive value of prognostic nutritional index(PNI) and lymphocyte-monocyte ratio(LMR) in severe radiotherapy-induced oral mucositis(RIOM) during treatment of patients with head and neck cancer,and to construct a risk prediction model and test the prediction effect. METHODS A total of 502 patients with head and neck cancer who underwent radiotherapy were recruited from September 2021 to October 2023 in Xiangya Hospital Central South University. The participants were randomly divided into training group and validation group at a ratio of 7:3. According to whether severe RIOM occurred,they were divided into severe RIOM group and non-severe RIOM group. Univariate analysis and logistic regression analysis were used to screen the risk factors of severe RIOM. The receiver operating characteristic(ROC) curve was used to evaluate its prediction effect and R4.3.2 software was used to draw nomograms and decision curve. RESULTS The risk prediction model for patients with head and neck cancer during treatment had five factors,including the number of comorbidities(OR=2.221,95％CI=1.185-4.165),surgical history(OR=2.938,95％CI=1.393-6.198),the degree of tumor differentiation(OR=1.511,95％CI=1.090-2.094),PNI(OR=0.892,95％CI=0.852-0.934),LMR(OR=0.512,95％CI=0.254-1.030). Model formula:Y=2.102+0.413×degree of differentiation+0.798×number of comorbidities+1.078×surgical history-0.114×PNI-0.669×LMR. The validation results of the prediction model showed that the area under the ROC curve of the training group was 0.847(P<0.001),the area under the curve of the validation group was 0.808(P<0.001),and the P values of the Hosmer-Lemeshow test of the modeling group and the validation group were both greater than 0.05. The decision curve was above the reference line within most of the high-risk thresholds. CONCLUSION The risk prediction model constructed in this study has good effect,which can predict the risk of severe RIOM during radiotherapy in patients with head and neck cancer,providing the reference for taking preventive intervention measures for high-risk patients.

Objective To investigate the prevalence and severity of symptoms and to construct symptom networks in head and neck cancer patients during treatment to identify core symptoms and symptom clusters.Methods 366 patients who were hospitalized in 3 tertiary hospitals in Changsha were selected using convenience sampling from March to October 2022 and asked to complete the M.D.Anderson Symptom Inventory-Head & Neck.Exploratory factors analysis was used to extract the symptom clusters,and R packages were used to construct the symptom severity network and symptom clusters network.The centrality indexes of the networks,including strength,closeness,and betweenness,were analyzed to identify core symptoms and core symptom cluster.Results The most common symptoms in head and neck cancer patients during treatment were dry mouth(93.44％),fatigue(89.07％),loss of appetite(86.34％),and difficulty swallowing or chewing(85.79％),and the most severe symptoms were dry mouth,loss of appetite,oral or pharyngeal mucus,and difficulty swallowing or chewing.4 symptom clusters were extracted,namely oral-pharyngeal,gastrointestinal,emotional-sleep,and sickness-sensing behavioral,which could explain 67.415％of the total variance.In the symptom severity network,oral or pharyngeal mucus(rs=9.60)was a symptom with the highest strength.In the symptom clusters network,oral or pharyngeal mucus(rs=1.20),nausea(rs=1.00),fatigue(rs=1.10),and drowsiness(rs=0.97)were the symptoms with the highest strength across 4 symptom clusters.Conclusion Oral or pharyngeal mucus,nausea,fatigue,and drowsiness are the core symptoms of symptom clusters in head and neck cancer patients during treatment.Oral-pharyngeal symptom cluster is the core symptom cluster.It is recommended that clinical staff should develop interventions based on the core symptoms and symptom cluster to implement precise symptom management and improve symptom management efficiency.

Objective:To explore the clinicopathological characteristics and the related influencing factors of efficacy and prognosis of elderly patients with malignant mesothelioma(MM)in Chinese population.Methods:We retrospectively analyzed the clinical data of 115 patients aged 65 years and above who were diagnosed with MM in Beijing Hospital, Peking University Cancer Hospital, and Cancer Hospital of Chinese Academy of Medical Sciences between November 2007 and July 2024, and the patients were grouped according to age(≥75 years in the older group and <75 years in the younger group), histological types and therapy regimens.Kaplan-Meier curve and Log-rank test were performed.Cox regression was used in prognostic analysis.Results:The positive expression rate of Calretinin in the Chinese elderly population with MM was consistent with previous reports, while the positive rates of Cytokeratin 5/6(CK5/6), WT-1, and D2-40 were much lower.The overall response ratio(ORR)for first-line treatment was 17.3%(9/52), and the disease control rate(DCR)was 92.3%(48/52).The ORR for second-line treatment was 7.7%(1/13)and the DCR was 76.9%(10/13).The ORR and DCR were higher in the first-line immunotherapy group than in the chemotherapy group, 50.0% vs.14.6%( P=0.134)and 100.0% vs.91.6%( P=1.000), respectively.The ORR in the second-line immunotherapy group was higher than that in the chemotherapy group, 25.0% vs.0, respectively, and the DCR were both 75.0% in two groups.The median progression free survival(mPFS)was 9.2 months and median overall survival(mOS)was 19.0 months for patients receiving first-line treatment, and the mPFS was 3.3 months and mOS was 11.0 months for second-line therapy.The first-line immunotherapy provided more shorter mPFS(1.6 months vs.9.2 months, P=0.081)and longer mOS(not reached vs.18.1 months, P=0.147)than the chemotherapy group.The younger group had prolonged mPFS(9.7 months vs.7.2 months, P=0.305)while shorter mOS(18.1 months vs.23.9 months, P=0.289)compared with the older group, and none of them reached statistical differences.Both mPFS and mOS were prolonged in the epithelioid subtype compared with the non-epithelioid subtypes, 10.4 months vs.1.6 months( P<0.001)and 20.3 months vs.4.6 months( P=0.803), respectively.Both mPFS(7.1 months vs.4.7 months, P=0.583)and mOS(18.3 months vs.6.3 months, P=0.134)were prolonged in the second-line chemotherapy group compared with the immunotherapy group.The Cox regression analysis showed that gender, Eastern Cooperative Oncology Group, Performance Status(ECOG PS)and positive CK5/6 were both the independent predictors for the first-line PFS.Histological type was an independent prognostic factor for the first-line OS. Conclusions:MM in the Chinese elderly population exhibits unique clinicopathologic characteristics.The immunotherapy improves ORR, DCR and prolongs mOS in first-line use, and improves ORR in second-line.First-line treatment improves mPFS in the younger group compared with the older group.Multivariate Cox regression demonstrates that gender, ECOG PS and CK5/6 expression are both predictors of efficacy, and histological type is an independent prognostic factor for survival of the Chinese elderly population with MM.

Objective lo analyze the difference of peri-coronary tat attenuation index(pr Al)on different grades of hypertension(HT),and to explore the value of pFAI in evaluating the risk of HT patients.Methods Retrospective data on hospitalized patients who underwent coronary computed tomography angiography(CCTA)examination for chest pain were collected.A total of 415 clini-cally confirmed HT patients were selected as observation group(including 132 patients in grade 1 HT group,137 patients in grade 2 HT group,146 patients in grade 3 HT group),and 187 non-hypertension patients during the same period as control group.The differ-ence of fat attenuation index(FAI)in three main coronary arteries[left anterior descending artery(LAD),left circumflex artery(LCX),right coronary artery(RCA)]was compared,and the correlation between pF AI and HT patients was analyzed.Results RCA-FAI(-78.86 HU±7.66 HU)and LAD-FAI(-80.62 HU±7.50 HU)were higher in HT group than those in control group(-84.46 HU± 8.00 HU,-83.43 HU±7.51 HU,P＜0.05).pFAI value was higher in grade 3 HT group than that in grade 1 HT group and grade 2 HT group(P＜0.05),while there were no differences between grade 1 HT group and grade 2 HT group(P＞0.05).After adjusting the influence of traditional risk factors and coronaryartery disease,RCA-FAI had relatively closer relationship with HT grades(r=0.47,P＜0.001).Conclusion LAD-FAI,RCA-FAI in the HT group are higher than those in control group and RCA-FAI has relatively closer relationship with HT grades,suggesting that RCA-FAI may be an imaging indicator to evaluate the pro-gression of HT and predict the risk of HT.

Objective:This study aimed to construct a risk prediction model for obstructive sleep apnea（OSA） related hypertension based on the nomogram, and to explore the independent risk factors for OSA-related hypertension, so as to provide reference for clinical treatment decision-making. Methods:The clinical data of OSA patients diagnosed by polysomnography from October 2019 to December 2021 were collected retrospectively and randomly divided into training sets and validation sets. A total of 1 493 OSA patients with 27 variables were included. The least absolute shrinkage and selection operator（Lasso） logistic regression model was used to select potentially relevant features and establish a nomogram for OSA-related hypertension.The performance and clinical benefits of this nomogram were verified in terms of discrimination, calibration ability and clinical net benefit. Results:Multivariate logistic regression showed that body mass index（BMI）, family history of hypertension, lowest oxygen saturation（LSaO2）, age and cumulative percentage of total sleep time with oxygen saturation below 90% were independent risk factors for OSA-related hypertension. Lasso logistic regression identified BMI, family history of hypertension, LSaO2 and age as predictive factors for inclusion in the nomogram. The nomogram provided a favorable discrimination, with a C-indexes of 0.835（95% confidence interval[CI ]0.806-0.863） 0.865（95%CI 0.829-0.900） for the training and validation cohort, respectively, and well calibrated. The clinical decision curve analysis displayed that the nomogram was clinically useful. Conclusion:Compared with cumulative percentage of total sleep time with blood oxygen saturation below 90%, LSaO2 may have a greater impact on the incidence of OSA-related hypertension, and the effects of different times and degrees of hypoxia on OSA-related hypertension should be further explored in the future. Apnea hypopnea index involvement is weak in predicting OSA-related hypertension, and the blood oxygen index may be a better predictor variable. Furthermore, we established a risk prediction model for OSA-related hypertension patients using nomogram, and demonstrated that this prediction model was helpful to identify high-risk OSA-related hypertension patients. This model can provide early and individualized diagnosis and treatment plans, protect patients from the serious.
Humans ; Sleep Apnea, Obstructive/complications* ; Nomograms ; Hypertension/epidemiology* ; Male ; Female ; Risk Factors ; Middle Aged ; Retrospective Studies ; Polysomnography ; Logistic Models ; Body Mass Index ; Adult

Objective:To construct the evaluation system of Intensive Care Unit (ICU) practice base for master of nursing specialist.Methods:The evaluation indicators at all levels were initially formulated through literature review and semi-structured interviews. Using the sampling method, 25 experts were selected from August to October 2021 for correspondence. The evaluation system of ICU practice base for master of nursing specialist was finally determined by Delphi method and analytic hierarchy process.Results:A total of two rounds of consultation were conducted. The expert positive coefficients were 88% (22/25) and 91% (20/22), the expert authority coefficients were 0.953 and 0.948, and the Kendall's W were 0.211 and 0.210 respectively with statistical differences ( P<0.01). The final evaluation system of ICU practice base for master of nursing specialist included 4 first-level indicators, 15 second-level indicators and 103 third-level indicators and evaluation contents. Conclusions:The evaluation system of ICU practice base for master of nursing specialist is scientific, reliable and comprehensive, which can provide reference for formulating the evaluation criteria of clinical practice base for nursing graduate students.