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.

ObjectiveGastrodia elata has evolved ecological types with shortened rhizome internodes and diversified flower and fruit coloration in response to different altitudes. Studying the genetic mechanisms of different ecotype germplasm is significant for guiding variety breeding in different cultivation areas. MethodsThe bHLH gene family was identified based on the whole-genome datasets of G. elata f. elata and G. elata f. glauca. Subsequently， the gene family members were subject to analysis， including gene structure， chromosomal localization， cis-acting elements， gene synteny， and phylogeny. Combined with transcriptome data and quantitative Real-time PCR， the expression patterns of bHLH genes in the stems of the different G. elata ecotype germplasm were analyzed. Finally， correlation analysis was conducted between gene expression patterns and color to obtain the key bHLH genes regulating the color formation of stem. ResultsA total of 63 bHLH genes were identified in both G elata f. elata and G. elata f. glauca， unevenly distributed across 17 chromosomes and clustered into 16 subfamilies， with significant expansion in some family members. Obvious inversions of bHLH genes on the same chromosome and interchromosomal translocations were detected in the two ecotype germplasm. Among these genes， 12 bHLH genes （such as bHLH62-3 and bHLH74） were associated with the bright yellow color of G elata f. elata stem， while 9 bHLH genes （such as PIL13， UNE12， and bHLH130） were correlated with the red color of G. elata f. glauca stem. Compared to G. elata f. glauca， the bHLH48 expression level was significantly higher in flowers and scale leaves of G elata f. elata， and the bHLH62-3 expression level was significantly higher in all organs of G elata f. elata. ConclusionsFunctional pathway divergence of the bHLH family members has occurred across different chromosomes in G elata f. elata and G. elata f. glauca. Through synergism or antagonism with other genes， 21 bHLH genes participate in the coloration metabolic pathway regulation of stems， flowers， and fruits. Specifically， bHLH62-3 is involved in regulating stem color differentiation in the anthocyanin biosynthesis pathway of G. elata， thus relevant to the color formation of stem. Additionally， GebHLH48 positively regulates flowering-related pathways to promote the early-flowering phenotype of G. elata f. elata. These findings have laid the foundation for analyzing the genetic regulatory mechanisms underlying the color formation of the G. elata stem.

ObjectiveGastrodia elata has evolved ecological types with shortened rhizome internodes and diversified flower and fruit coloration in response to different altitudes. Studying the genetic mechanisms of different ecotype germplasm is significant for guiding variety breeding in different cultivation areas. MethodsThe bHLH gene family was identified based on the whole-genome datasets of G. elata f. elata and G. elata f. glauca. Subsequently， the gene family members were subject to analysis， including gene structure， chromosomal localization， cis-acting elements， gene synteny， and phylogeny. Combined with transcriptome data and quantitative Real-time PCR， the expression patterns of bHLH genes in the stems of the different G. elata ecotype germplasm were analyzed. Finally， correlation analysis was conducted between gene expression patterns and color to obtain the key bHLH genes regulating the color formation of stem. ResultsA total of 63 bHLH genes were identified in both G elata f. elata and G. elata f. glauca， unevenly distributed across 17 chromosomes and clustered into 16 subfamilies， with significant expansion in some family members. Obvious inversions of bHLH genes on the same chromosome and interchromosomal translocations were detected in the two ecotype germplasm. Among these genes， 12 bHLH genes （such as bHLH62-3 and bHLH74） were associated with the bright yellow color of G elata f. elata stem， while 9 bHLH genes （such as PIL13， UNE12， and bHLH130） were correlated with the red color of G. elata f. glauca stem. Compared to G. elata f. glauca， the bHLH48 expression level was significantly higher in flowers and scale leaves of G elata f. elata， and the bHLH62-3 expression level was significantly higher in all organs of G elata f. elata. ConclusionsFunctional pathway divergence of the bHLH family members has occurred across different chromosomes in G elata f. elata and G. elata f. glauca. Through synergism or antagonism with other genes， 21 bHLH genes participate in the coloration metabolic pathway regulation of stems， flowers， and fruits. Specifically， bHLH62-3 is involved in regulating stem color differentiation in the anthocyanin biosynthesis pathway of G. elata， thus relevant to the color formation of stem. Additionally， GebHLH48 positively regulates flowering-related pathways to promote the early-flowering phenotype of G. elata f. elata. These findings have laid the foundation for analyzing the genetic regulatory mechanisms underlying the color formation of the G. elata stem.

ObjectiveTo observe the clinical efficacy and safety of Tangning Tongluo tablets in the treatment of nonproliferative diabetic retinopathy （DR）. MethodsFourteen research centers participated in this study， which spanned a time interval from September 2021 to May 2023. A total of 240 patients with nonproliferative DR were included and randomly assigned into an observation group （120 cases） and a control group （120 cases）. The observation group was treated with Tangning Tongluo tablets， and the control group with calcium dobesilate capsules. Both groups were treated for 24 consecutive weeks. The vision， DR progression rate， retinal microhemangioma， hemorrhage area， exudation area， glycosylated hemoglobin （HbA1c） level， and TCM syndrome score were assessed before and after treatment， and the safety was observed. ResultsThe vision changed in both groups after treatment （P<0.05）， and the observation group showed higher best corrected visual acuity （BCVA） than the control group （P<0.05）. The DR progression was slow with similar rates in the two groups. The fundus hemorrhage area and exudation area did not change significantly after treatment in both groups， while the observation group outperformed the control group in reducing the fundus hemorrhage area and exudation area. There was no significant difference in the number of microhemangiomas between the two groups before treatment. After treatment， the number of microhemangiomas decreased in both the observation group （Z=-1.437， P<0.05） and the control group （Z=-2.238， P<0.05）， and it showed no significant difference between the two groups. As the treatment time prolonged， the number of microhemangiomas gradually decreased in both groups. There was no significant difference in the HbA1c level between the two groups before treatment. After treatment， the decline in the HbA1c level showed no significant difference between the two groups. The TCM syndrome score did not have a statistically significant difference between the two groups before treatment. After treatment， neither the TCM syndrome score nor the response rate had significant difference between the two groups. With the extension of the treatment time， both groups showed amelioration of TCM syndrome compared with the baseline. ConclusionTangning Tongluo tablets are safe and effective in the treatment of nonproliferative DR， being capable of improving vision and reducing hemorrhage and exudation in the fundus.

OBJECTIVE To explore the time windows for postoperative pneumonia in patients undergoing different surgeries,providing evidence-based references for optimizing infection monitoring and prevention and control strategies.METHODS Sociodemographic characteristics,clinical information and surgical details of 263 patients with postoperative pneumonia from four different types of medical institutions between Jan.2019 and Dec.2024 were retrospectively collected.The time windows for postoperative pneumonia in patients undergoing different surgeries were analyzed.RESULTS There were no statistically significant differences in the time windows for post-operative pneumonia among groups in terms of sociodemographic factors and underlying diseases.However,sta-tistically significant differences were observed in the time windows for postoperative pneumonia based on surgery type,incision type,surgical approach and surgery duration(P＜0.05).The average time for the onset of postop-erative pneumonia in 263 patients was 2.00(1.00,7.00)days.The postoperative time windows varied for sur-geries involving different systems.The peak incidence occurred on day 0(16 cases)and day 1(17 cases)af-ter neurological surgery,while the peak incidence for digestive system and orthopedic surgeries was on day 1.The time span for the onset of pneumonia after skin surgeries was wider(0-53 days postoperatively)without a clear peak.In addition,33.33％of cardiovascular system surgery cases developed pneumonia 10 days postoperatively.There were also significant time differences in the diagnostic elements of postoperative pneumonia,with fever and abnormal white blood cell counts appearing earlier(median appearance time length:4.00 days)than lung imaging changes(median appearance time length:7.00 days).CONCLUSIONS This study demonstrates significant time differences in the on-set of postoperative pneumonia and confirms the significant spatiotemporal heterogeneity in the diagnostic elements of postoperative pneumonia.These findings provide a quantitative basis for developing dynamic,surgery-type-spe-cific monitoring protocols and prevention and control measures for postoperative pneumonia.

Objective:To analyze the differences in heart size，shape and function between fetuses with tetralogy of Fallot（TOF）and fetuses with malalignment ventricular septal defect（VSD）and pulmonary stenosis（PS）（named as，mild TOF）by fetal heart quantification（fetal HQ）and multi-parameter fetal echocardiography.Methods:From June 2021 to June 2023，50 fetuses with TOF（TOF group）diagnosed by fetal echocardiography at the Department of Diagnostic Ultrasound & Echocardiography Sir Run Run Shaw Hospital，Zhejiang University College of Medicine and 34 fetuses with VSD and PS matched to gestational age（mild TOF group）were retrospectively selected. Cardiovascular parameters were measured by 2D echocardiography，M-mode echocardiography and fetal HQ，including aortic dimension（AO）and its Z-score（AO Z-score），pulmonary artery diameter（PA）and its Z-score（PA Z-score），PA/AO ratio，main pulmonary artery diameter（MPA）and its Z-score（MPA Z-score），left pulmonary artery diameter（LPA）and its Z-score（LPA Z-score），right pulmonary artery diameter（RPA）and its Z-score（RPA Z-score），McGoon index（MGI），tricuspid annular diameter（TV）and its Z-score（TV Z-score），mitral annular diameter（MV）and its Z-score（MV Z-score），and MV/TV ratio. Measurements of global ventricular morphologic parameters included cardiac axis，left and right ventricular transverse diameters（LVW，RVW）and their ratio（LVW/RVW），left and right ventricular long diameters（LVL，RVL）and LVL/RVL ratio，left and right ventricular areas（LVA，RVA）and LVA/RVA ratio，global spherical index（GSI）and its Z-score，four chamber view transverse diameter（4CV-Width-ED），four chamber view end-diastolic longitudinal diameter（4CV-Length-ED），four chamber view end-diastolic area（4CV-Area-ED）. Measurements of left and right global ventricular functional parameters included fractional left ventricular and right ventricular area changes（LVFAC，RVFAC），left ventricular ejection fraction（LVEF），left ventricular and right ventricular global longitudinal strains（LVGLS，RVGLS）；end diastolic width diameter（Width-ED），SI and fractional shortening rate（FS）of 24 segments of left and right ventricles. The differences of the above parameters between the two groups were analyzed and compared，and the relationships between absolute values of GLS in left and right ventricles of TOF fetus and PA Z-score，MPA Z-score and PA/AO were analyzed. Binary Logistic regression model was used to select the best variables，and ROC curve was adopted to analyze the predictive values of ultrasonic parameter variables on TOF.Results:There were statistically significant differences in MV，TV，RVW，RVL，LVL，LVA，RVA，4CV-Length-ED，LVGLS，RVGLS，RVFAC，PA/AO，MPA，RPA，LPA，MGI，PA and PA Z-score between TOF group and mild TOF group（all P<0.05）. The Width-ED values of all segments of left ventricle were statistically different between TOF group and mild TOF group（all P<0.05）. There were statistical differences in SI between LV segments 4~6 and 8~24，and RV segments 1~21（all P<0.05）. The FS values between the 8th and 10th to 18th segments of RV revealed statistical differences（all P<0.05）. There were no statistical differences in the other parameters（all P>0.05）. The absolute value of LVGLS in TOF fetuses was positively correlated with PA Z-score，MPA Z-score and PA/AO（ r = 0.313，0.344，0.304，all P<0.05），and the absolute value of RVGLS was positively correlated with PA Z-score，MPA Z-score，and PA/AO（ r = 0.323，0.334，0.357，all P<0.05）. Binary Logistic regression model analysis confirmed that LVGLS，RVFAC and PA/AO were predictive variables. ROC curve analysis showed that the areas under the curves of LVGLS，RVFAC and PA/AO for predicting TOF were 0.746，0.693 and 0.849 respectively. The combined prediction efficiency was higher，and the area under the curve was 0.906. Conclusions:Fetal HQ combined with multiple fetal echocardiographic quantification indices can evaluate the differences in fetal heart size，shape and function between TOF and mild TOF. It is expected to provide important reference information in prenatal diagnosis and consultation for fetuses with TOF and mild TOF.

Objective:To systematically analyze the risk factors for knee injury complicated with adolescent tibial tubercle fractures (ATTF).Methods:A retrospective study was conducted to analyze the clinical data of the 90 adolescent patients (the observation group) who had been treated and fully followed up for knee injury complicated with ATTF at Department of Pediatric Orthopedics, The Second General Hospital of Fuzhou from September 2017 to April 2024. There were 86 males and 4 females, with a mean age of 14.0 (13.0, 14.0) years. The injury was on the left side in 51 cases, on the right side in 34 ones, and on the bilateral sides in 5 ones. All their injuries resulted from sports activities. In addition, another contemporary 90 patients with knee injury but no exercise-induced ATTF were enrolled as the control group, including 54 males and 36 females with a mean age of 14.0 (13.0, 14.3) years. Their injury was on the left side in 45 cases, on the right side in 41 ones, and on the bilateral sides in 4 ones. Univariate analysis was conducted to compare gender, age, injury side, height, weight, body mass index, history of Osgood-Schlatter disease (OSD), and injury mechanism between the 2 groups. Variables with P<0.1 were included in a multivariate logistic regression analysis to screen independent risk factors for ATTF. The receiver operating characteristic (ROC) curve was used to evaluate their predictive values. The optimal cutoff value was determined by Youden index. Results:The univariate analysis showed that gender, height, weight, body mass index, and OSD history were significantly correlated with the occurrence of ATTF ( P<0.05). The multivariate logistic regression analysis further confirmed that male ( P=0.017), height ( P=0.021), weight ( P=0.014), body mass index ( P=0.032) and history of OSD ( P=0.003) were independent risk factors for ATTF. The ROC curve analysis showed that weight had the largest area under the curve (AUC) in prediction of the ATTF risk, suggesting that its predictive value was the most significant. Conclusion:Male, height, weight, body mass index, and prior OSD are independent risk factors for ATTF, with the highest predictive value in weight.

Objective The purpose of this study was to explore the application effect of BOPPPS combined with case simulation teaching method in nursing risk education of nursing students.By comparing the traditional teaching methods,the in-fluence of BOPPPS combined with case simulation teaching method on nursing students' occupational risk cognition ability,pa-tient safety perception ability and teaching satisfaction was evaluated.Methods 100 cases of nursing students in geriatric oncol-ogy department were divided into observation group and control group,50 cases in each group.The control group used traditional teaching methods;the observation group adopted the teaching mode of BOPPPS combined with case teaching method.The occu-pational risk cognition ability,patient safety perception ability and satisfaction with teaching mode were compared between the two groups.Results The occupational risk cognitive ability,patient safety perception ability and teaching satisfaction of the observa-tion group were better than those of the control group(P＜0.05).Conclusion The application of BOPPPS combined with case simulation teaching method in nursing risk teaching of nursing students can improve their occupational risk cognition ability,pa-tient safety perception ability and teaching satisfaction.

Objective To analyze the clinical and immunological characteristics of a rare case of CHARGE syndrome,we summarize the genotype and phenotype in the Chinese patient population,and explore the underlying immunopathogenic mechanisms.Methods Clinical data from a pediatric patient with CHARGE syndrome were collected and analyzed.A comprehensive analysis of the Chinese patient population was conducted.Gene analysis and immunological characterization were performed using flow cytometry,deep sequencing,and quantitative PCR.Results The proband was a premature female infant whose primary clinical manifestations included congenital heart disease,recurrent respiratory infections,respiratory failure,airway dysplasia,hearing impairment,and bilateral choroidal coloboma.Whole-exome sequencing revealed a de novo heterozygous nonsense mutation in the CHD7 gene,c.5122C＞T(p.Gln1708Ter),classified as pathogenic according to ACMG criteria.Immunological studies indicated impaired thymic output of T cells,significant alterations in the number and proportion of CD8+T cell subsets,increased apoptosis,and defective activation and production of key effector cytokines such as IFN-γ by CD8+T cells.However,no significant abnormalities were observed in peripheral lymphocyte proliferation.Conclusion CHARGE syndrome is a rare autosomal dominant genetic disorder primarily caused by mutations in the CHD7 gene.The main clinical features include ocular defects,cardiac disease,choanal atresia/cleft lip and palate,growth retardation,gonadal hypoplasia,and ear anomalies.This case study suggests that CHARGE syndrome is associated with abnormalities in the development,apoptosis,and effector functions of immune cells.

Objective To investigate the awareness of female patients with pelvic floor dysfunction regarding minimally invasive laser treatment of the reproductive tract and analyze the factors influencing their decision-making intentions,this study aims to provide a foundation for early treatment of pelvic floor dysfunction and further development in reproductive health management.Methods A convenience sampling method was employed to select 164 female patients with pelvic floor dysfunction who sought treatment at the Pelvic Rehabilitation Center of Dongguan Maternal and Child Health Care Hospital between June 2023 and August 2024.The study utilized the Female Sexual Function Index,Incontinence Quality of Life Questionnaire,and Family Support Self-Assessment Scale to conduct a survey.Binary logistic stepwise regression analysis was conducted to investigate the factors influ-encing patients'inclination towards undergoing genital laser minimally invasive treatment.Results Among the 164 female patients,143(87.2%)expressed an intention to receive treatment,with 22.6%demonstrating a rela-tively clear understanding of genital laser minimally invasive treatment.Logistic regression analysis revealed that occupation significantly influenced treatment intention(P<0.05).Compared to healthcare professionals,individuals in the teaching profession(OR=10.81,95%CI:1.04～112.21),self-employed individuals(OR=20.34,95%CI:3.46～119.43),and those in other professions(OR=16.26,95%CI:4.05～65.29)were more inclined to express willingness for undergoing treatment.Furthermore,a lower score on the Incontinence Quality of Life scale was found to positively correlate with treatment intention(OR=0.96,95%CI:0.93～0.99).Conclusion Although patients express a high intention to undergo minimally invasive genital laser treatment,their overall awareness of the procedure remains insufficient.