Objective To elucidate the molecular mechanism of sirtuin-3 (SIRT3) in regulating mitochondrial biogenesis in human renal tubular epithelial cells. Methods Cells were stimulated with different concentrations of H₂O₂ and divided into four groups: control (NC), 50 μmol/L H₂O₂, 110 μmol/L H₂O₂ and 150 μmol/L H₂O₂. SIRT3 protein expression was then measured. SIRT3 was knocked down with siRNA, and cells were further assigned to five groups: control (NC), negative-control siRNA (NCsi), SIRT3-siRNA (siSIRT3), NCsi+H₂O₂, and siSIRT3+H₂O₂. After 24 h, cellular adenosine triphosphate (ATP) and mitochondrial superoxide anion (O₂•⁻) levels were determined, together with mitochondrial expression of SIRT3, peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α), nuclear respiratory factor 1 (NRF1), mitochondrial transcription factor A (TFAM), superoxide dismutase 2 (SOD2), acetylated-SOD2 and adenosine monophosphate activated protein kinase α1 (AMPKα1). Results The 110 and 150 μmol/L H₂O₂ decreased SIRT3 protein (both P<0.05). ATP and mitochondrial O₂•⁻ did not differ between NC and NCsi groups (both P>0.05). Compared to the NCsi group, the siSIRT3 group exhibited elevated O₂•⁻ level, decreased SIRT3 protein and increased expression levels of SOD2 and acetylated SOD2 protein (all P<0.05). Compared to the NCsi group, the NCsi+H₂O₂ group exhibited decreased cellular ATP levels, elevated mitochondrial O₂•⁻ levels, and reduced protein expression levels of SIRT3, SOD2, TFAM, AMPKα1, PGC-1α and NRF1 (all P<0.05). Compared with the siSIRT3 group, the siSIRT3+H₂O₂ group showed a decrease in cellular ATP levels, an increase in mitochondrial O₂•⁻ levels, a decrease in SIRT3, SOD2, TFAM, AMPKα1, PGC-1α and NRF1 protein expression levels and a decrease in acetylated SOD2 protein expression levels (all P<0.05). Compared with the NCsi+H₂O₂ group, the siSIRT3+H₂O₂ group showed a decrease in cellular ATP levels, an increase in mitochondrial O₂•⁻ levels, a decrease in SIRT3, AMPKα1, PGC-1α and NRF1, TFAM protein expression levels, and an increase in SOD2 and acetylated SOD2 protein expression levels (all P<0.05). Conclusions SIRT3 promotes mitochondrial biogenesis in tubular epithelial cells via the AMPK/PGC-1α/NRF1/TFAM axis, representing a key mechanism through which SIRT3 ameliorates oxidative stress-induced mitochondrial dysfunction.

BackgroundClinical supervision is a critical component in the training and professional development of psychological counselors. Scientifically evaluating the effectiveness of clinical supervision is essential， yet reliable and effective tools for such assessments are lacing in China. ObjectiveTo translate Supervision Evaluation and Supervision Competence （SE-SC） Scale into Chinese version and evaluate its reliability and validity in clinical supervision in China， so as to provide a tool for the evaluation of supervisory effectiveness. MethodsThe SE-SC scale was translated， back-translated and culturally adapted， followed by a pilot survey to develop the Chinese version of SE-SC scale. A total of 42 counselors engaged in clinical counseling and receiving supervision at a counseling center in Shanghai from July 2021 to February 2022 were selected as the study participants. Item analysis was conducted to assess item discrimination， with critical ratio method applied to determine which items retention. Hierarchical cluster analysis was performed to compare the structure of Chinese version with the original scale. Criterion-related validity and convergent validity were used to evaluate the validity of the scale， while Cronbach's α coefficient was used to assess its reliability. ResultsChinese version of SE-SC scale consisted of a total of 28 item， including six clusters. Registered supervisors scored significantly higher than internship supervisors on the total score and on clusters three， four， five and six （t=2.536， 2.747， 5.881， 3.718， 6.090， P<0.05）. The total and cluster scores of the Chinese version of the SE-SC scale were positively correlated with self-rated supervision helpfulness and overall satisfaction （r=0.492~0.758， 0.412~0.815， P<0.01）. The Cronbach's α coefficient for the overall scale was 0.975，with values for the six clusters were 0.938， 0.821， 0.962， 0.871， 0.884 and 0.823， respectively. ConclusionChinese version of SE-SC scale demonstrates good reliability and validity， and it can be considered as a promising assessment tool for evaluating the effectiveness of clinical supervision.

ObjectiveTo investigate the main risk factors for liver cirrhosis in chronic hepatitis B （CHB） patients with high metabolic risk， to establish a noninvasive predictive model， and to compare the diagnostic efficiency of this model and other models including fibrosis-4 （FIB-4）， aspartate aminotransferase-to-platelet ratio index （APRI）， gamma-glutamyl transpeptidase-to-platelet ratio （GPR）， and Forns index. MethodsA total of 527 CHB patients with high metabolic risks who were admitted to The Second Affiliated Hospital of Guangxi Medical University from September 1， 2017 to October 31， 2022 were enrolled as subjects， and they were randomly divided into modeling group with 368 patients and validation group with 159 patients at a ratio of 7∶3. The LASSO regression analysis and the multivariate Logistic regression analysis were performed for the modeling group to identify independent risk factors， and a nomogram model was established. The receiver operating characteristic （ROC） curve， the calibration curve， and the decision curve analysis were used to validate the nomogram prediction model in the modeling group and the validation group and assess its discriminatory ability， calibration， and clinical practicability. The Delong test was used to compare the area under the ROC curve （AUC） of the nomogram prediction model and other models. ResultsThe multivariate Logistic regression analysis showed that prealbumin （odds ratio ［OR］ = 0.993， 95% confidence interval ［CI］： 0.988 — 0.999， P= 0.019）， thrombin time （OR=1.182， 95% CI： 1.006 — 1.385， P=0.047）， log₁₀ total bilirubin （TBil） （OR=1.710， 95%CI： 1.239 — 2.419， P=0.001）， and log₁₀ alpha-fetoprotein （AFP） （OR=1.327， 95%CI： 1.052 — 1.683， P=0.018） were independent influencing factors for liver cirrhosis in CHB patients with high metabolic risks. A nomogram model for risk prediction was established based on the multivariate analysis， which had an AUC of 0.837 （95%CI： 0.788 — 0.888）， a specificity of 73.5%， and a sensitivity of 84.7%， as well as a significantly higher diagnostic efficiency than the models of FIB-4 （0.739）， APRI （0.802）， GPR （0.800）， and Forns index （0.709） （Z=2.815， 2.271， 1.989， and 2.722， P=0.005， 0.017， 0.045， and 0.006）. ConclusionThe nomogram model established based on prealbumin， thrombin time， log₁₀ TBil， and log₁₀ AFP has a certain clinical application value.

Objective: To explore the effects of Cldn14 gene knockout on renal metabolism and stone formation in rats，so as to provide reference for research in the field of urinary calium metabolism and stone formation. Methods: Cldn14 gene knockout homozygous rats and wild-type rats of the same age were randomly divided into 4 groups：wild-type control (WC) group，wild-type ethylene glycol (WE) group，gene knockout control (KC) group and gene knockout ethylene glycol (KE) group，with 10 rats in each group.The WE and KE groups were induced with ethylene glycol + ammonium chloride to form kidney stones，while the WC and KC groups received normal saline gavage.After 4 weeks of standard maintenance feeding，the urine samples were collected to detect the venous blood.The kidneys were collected for HE，Pizzolatto's staining and transmission electron microscopy.The protein in renal tissues was extracted to detect the expressions of Claudin16 and Claudin19. Results: Crystal deposition was observed in the renal tubular lumen of the WE and the KE groups，and more crystals were detected in the KE group.The WE group had a large number of intracytoplasmic black crystalline inclusions observed in renal tubular epithelial cells under transmission electron microscope，followed by the KE and KC groups.Compared with WC and WE groups，KC and KE groups had significantly decreased serum calcium and magnesium levels but significantly increased urinary calcium level.In addition，the urinary calcium level was higher in the WE group than in the WC group and higher in the KE group than in the KC group.The KE group had lower level of Claudin16，but there was no significant difference in the level of Claudin19 among the 4 groups(P>0.05). Conclusion: Knockout of Cldn14 gene alone cannot effectively reduce urinary calcium excretion or reduce the risk of stone formation in rats，which may be related to the decrease of Claudin16 level.

ObjectiveThe U6 promoter is an essential element for driving sgRNA expression in the clustered regularly interspaced short palindromic repeat sequences/CRISPR-associated protein 9（CRISPR/Cas9）gene editing system in dicotyledonous plants. Endogenous U6 promoters typically exhibit higher transcriptional activity， which can significantly improve gene editing efficiency. This study aims to identify endogenous U6 promoters in Artemisia annua to optimize its CRISPR/Cas9 gene editing system， which holds significant importance for its molecular breeding. MethodsOn the basis of the highly conserved U6 snRNA sequences in Arabidopsis thaliana， endogenous U6 promoters were screened in the A. annua genome. Expression vectors were constructed with candidate AaU6 promoter driving the firefly luciferase （LUC） reporter gene， and then transiently transformed into Nicotiana benthamiana. Transcriptional activities of the promoters were measured and compared by in vivo imaging and the Dual Luciferase Reporter assay. ResultsEight endogenous U6 promoters were successfully cloned from A. annua. Sequences alignment revealed that all these promoters contained the two conserved cis-acting elements， upstream sequence element （USE） and TATA-box， which affected their transcriptional activity. Dual-luciferase activity assays indicated that the transcriptional activities of AaU6-3， AaU6-1， and AaU6-5 were significantly higher than that of the Arabidopsis AtU6-26 promoter， with AaU6-3 exhibiting the highest activity. ConclusionThis study identified three endogenous AaU6 promoters with high transcriptional activity in A. annua， providing key functional elements for establishing an efficient gene editing system in A. annua. These findings will contribute to advancing precision molecular breeding and high-quality germplasm innovation in A. annua.

OBJECTIVE: This study aimed to identify high-risk areas for type 2 diabetes mellitus (T2DM) mortality to provide relevant evidence for interventions in emerging economies. METHODS: Empirical Bayesian Kriging and a discrete Poisson space-time scan statistic were applied to identify the spatiotemporal clusters of T2DM mortality. The relationships between economic factors, air pollutants, and the mortality risk of T2DM were assessed using regression analysis and the Poisson Log-linear Model. RESULTS: A coastal district in East Guangdong, China, had the highest risk (Relative Risk [RR] = 4.58, P < 0.01), followed by the 10 coastal districts/counties in West Guangdong, China (RR = 2.88, P < 0.01). The coastal county in the Pearl River Delta, China (RR = 2.24, P < 0.01), had the third-highest risk. The remaining risk areas were two coastal counties in East Guangdong, 16 districts/counties in the Pearl River Delta, and two counties in North Guangdong, China. Mortality due to T2DM was associated with gross domestic product per capita (GDP per capita). In pilot assessments, T2DM mortality was significantly associated with carbon monoxide. CONCLUSION High mortality from T2DM occurred in the coastal areas of East and West Guangdong, especially where the economy was progressing towards the upper middle-income level.
Diabetes Mellitus, Type 2/epidemiology* ; China/epidemiology* ; Humans ; Risk Factors ; Spatio-Temporal Analysis ; Air Pollutants/analysis* ; Socioeconomic Factors ; Bayes Theorem ; Female ; Male ; Middle Aged

Dental treatments for children and adolescents have unique clinical characteristics that differ from dental care for adults in terms of children's physiology, psychology, and behavior. These differences impose specific requirements on the application of local anesthesia in pediatric dental procedures. This article presents expert consensus on the principles of local anesthesia techniques in pediatric dental therapies, including the use of common anesthetic drugs and dosage control, safety and efficacy evaluation, and prevention and management of complications. The aim is to improve the safety and quality of pediatric dental treatments and offer guidance for clinical application by dentists.
Humans ; Child ; Anesthesia, Local/methods* ; Consensus ; Anesthesia, Dental/methods* ; Adolescent ; Anesthetics, Local/administration & dosage* ; Dental Care for Children

Due to the wide application of silver-containing dressings and silver-coated medical devices in clinical treatment; the extensive use of antibacterial agents and heavy metal agents in feed factories, Escherichia coli has formed the tolerance to silver ions. To systematically understand the known silver ion resistance mechanisms of E. coli, this article reviews the complex regulatory network and various physiological mechanisms of silver ion tolerance in E. coli, including the regulation of outer membrane porins, energy metabolism modulation, the role of efflux systems, motility regulation, and silver ion reduction. E. coli reduces the influx of silver ions by missing or mutating outer membrane porins such as OmpR, OmpC, and OmpF. It adapts to high concentrations of silver ions by altering the expression of ArcA/B and enhances the efflux capacity of silver ions under high-concentration silver stress via the endogenous Cus system and exogenous Sil system. Furthermore, the motility of bacteria is related to silver tolerance. E. coli has the ability to reduce silver ions, thereby alleviating the oxidative stress induced by silver ions. These findings provide a new perspective for understanding the formation and spread of bacterial tolerance and provide directions for the development of next-generation silver-based antimicrobials and therapies.
Escherichia coli/genetics* ; Silver/pharmacology* ; Drug Resistance, Bacterial ; Anti-Bacterial Agents/pharmacology* ; Porins/metabolism*

Objective To investigate the impact of adequate lumen preparation on the 12-month primary patency rate of drug-eluting stents(DES)in patients with lower extremity arteriosclerosis obliterans(ASO)and to identify risk and protective factors for in-stent restenosis.Methods A retrospective cohort study was conducted on 141 patients who underwent DES implantation for ASO at the Department of Vascular Surgery of the First Affiliated Hospital of Chongqing Medical University between April 2022 and April 2024.According to post-percutaneous transluminal angioplasty(PTA)dissection grade[severe dissection is defined as type C or higher based on National Heart,Lung,and Blood Institute(NHLBI)criteria]and residual stenosis rate(severe stenosis is defined as≥30%),the patients were categorized into an adequate preparation group(no severe dissection or severe residual stenosis;n=59)and an inadequate preparation group(n=82).DES primary patency at 12 months was assessed by color Doppler ultrasonography or angiography,defined as a peak systolic velocity ratio≤2.4 on duplex ultrasound or angiographic stenosis<50%.Kaplan-Meier analysis was used to compare intergroup differences in DES patency rate and freedom from clinically driven target lesion revascularization(CD-TLR).Multivariate logistic regression analysis was employed to identify risk and protective factors for DES restenosis and to evaluate the risk contributions of post-PTA dissection and residual stenosis to DES restenosis.Results Post-PTA,vascular dissection occurred in 94.3%(133/141)of treated limbs,among which 42.1%(56/133)were severe dissections,and severe residual stenosis was present in 44.0%(62/141)of limbs.Kaplan-Meier analysis revealed that the 12-month primary patency rate was significantly higher in the adequate preparation group than the inadequate group(HR=0.322,95%CI:0.132～0.789,P=0.013),while no significant difference was found in the rate of freedom from CD-TLR(HR=0.608,95%CI:0.187～1.937,P=0.407).Multivariate logistic regression analysis identified adequate lumen preparation(OR=0.228,95%CI:0.069～0.747,P=0.015)and Rutherford category 2～3(OR=0.205,95%CI:0.058～0.725,P=0.014)as independent protective factors against DES restenosis.The patency rate in the subgroup with coexisting severe dissection and residual stenosis was significantly lower than that in the uncomplicated group(55.6%vs 89.8%,P<0.001),with a 7.067-fold increased risk of re-stenosis(95%CI:2.424～20.602,P<0.001).Conclusion Adequate lumen preparation significantly improves the 12-month primary patency rate of DES.It,along with Rutherford category 2～3,serves as an independent protective factor against in-stent restenosis.Concurrent severe dissection and significant residual stenosis should be actively avoided during the procedure.

An ion chromatography method was developed for detection of nine kinds of trace organic amines(Methylamine,dimethylamine,trimethylamine,ethylamine,diethylamine,triethylamine,n-propylamine,n-butylamine,and ethanolamine)and six kinds of trace water-soluble inorganic cations(Li+,Na+,NH4+,K+,Ca2+,and Mg2+)in atmospheric fine particulate matter(PM2.5)in this wok.Various chromatographic columns(IonPac CS12,IonPac CS17 and IonPac CS19)were compared in terms of their separation efficiency for target analytes,and IonPac CS19 column was ultimately selected.Through meticulous optimization of the column temperature,a low temperature condition of 20℃was found to achieve the highest separation efficiency(All are above 1),effectively separating all 15 kinds of target analytes.Under the optimal analytical conditions inculding methanesulfonic acid(MSA)as eluent,100 μL of injection volume,column temperature at 20℃and eluent at flow rate of 1 mL/min,the detection limits of this method ranged from 0.05 to 7.15 μg/L,the quantification limits were 0.16-23.82 μg/L,and the spiking recoveries were 84%-105%.The proposed method exhibited high accuracy and excellent reproducibility,and was suitable for concurrent analysis and measurement of organic amines and water-soluble inorganic cations in PM2.5.