[摘 要] 目的：探讨lncRNA核内富集丰富转录本1（NEAT1）调控miR-1287-5p/DNA损伤诱导转录本4（DDIT4）轴对卵巢癌SKOV3细胞增殖、凋亡和侵袭的影响及其机制。方法：收集2023年6月至2024年6月期间湖北医药学院附属随州医院手术切除的27例卵巢癌患者的癌及癌旁组织标本，以及正常人卵巢上皮细胞IOSE80和卵巢癌细胞系SKOV3、CAOV3和A2780，RT-qPCR检测卵巢癌组织与细胞中lncRNA NEAT1、miR-1287-5p及DDIT4 mRNA表达。将SKOV3细胞分为Ctrl组、si-NC组、si-NEAT1组、si-NEAT1 + anti-miR-NC组、si-NEAT1 + anti-miR-1287-5p组、si-NEAT1 + vector组、si-NEAT1 + OE-DDIT4组。CCK-8法、克隆形成实验、流式细胞术、Transwell实验分别检测各组细胞增殖、凋亡及侵袭能力，WB法检测细胞中DDIT4、细胞周期蛋白D1（cyclin D1）、p53、迁移侵袭增强子1（MIEN1）蛋白水平。双萤光素酶报告基因验证lncRNA NEAT1与miR-1287-5p/DDIT4靶向结合关系，RNA pull-down实验、RNA免疫沉淀实验分别验证lncRNA NEAT1与miR-1287-5p、miR-1287-5p与DDIT4的靶向结合关系。另设pcDNA组（转染空载体pcDNA3.1）和pc-NEAT1 组（转染pcDNA-NEAT1）以验证NEAT1过表达效应。结果：卵巢癌组织中lncRNA NEAT1、DDIT4 mRNA表达水平显著高于癌旁组织（P < 0.05），而miR-1287-5p表达显著低于癌旁组织（P < 0.05）。敲低lncRNA NEAT1后，与Ctrl组和si-NC组相比，si-NEAT1组lncRNA NEAT1表达、DDIT4 mRNA表达均显著降低（均P < 0.05），miR-1287-5p 表达显著升高（P < 0.05）；而过表达lncRNA NEAT1则下调miR-1287-5p表达并上调DDIT4表达，呈现与敲低实验相反的调控效应；敲低lncRNA NEAT1后，克隆形成数、细胞增殖活性、细胞侵袭数均显著降低（均P < 0.05），细胞凋亡率显著升高（P < 0.05）；DDIT4、cyclin D1、MIEN1蛋白表达均显著降低（均P < 0.05），p53蛋白表达显著升高（P < 0.05）。进一步实验证实，anti-miR-1287-5p或OE-DDIT4均可减弱si-NEAT1对SKOV3细胞增殖和侵袭的抑制作用，同时减弱其对细胞凋亡的促进作用。lncRNA NEAT1靶向调控miR-1287-5p/DDIT4。结论：lncRNA NEAT1通过靶向调控miR-1287-5p/DDIT4轴促进SKOV3细胞增殖和侵袭，抑制细胞凋亡。

ObjectiveTo investigate the prevalence and associated factors of malocclusion among preschool children in Fengxian District, Shanghai, and to provide a scientific basis for local medical and public health departments to formulate targeted interventions. MethodsA cross-sectional study was conducted from September to December 2024. A total of 1 172 children aged 3‒5 years were recruited from 10 kindergartens across 5 townships in Fengxian District, using stratified cluster random sampling method. Clinical examinations were conducted to record occlusal parameters such as the relationship of the second deciduous molars, canine teeth relationship, anterior overbite and overjet, anterior and posterior crossbite, and crowded dentition. A questionnaire survey was performed to the parents to collect information on factors influencing malocclusion, including genetic predisposition, poor oral habits, dietary patterns, and sleep-related breathing conditions. Data were statistically analyzed using IBM SPSS Statistics 27 software, with chi-square test for categorical data and binary logistic regression analysis for influencing factors. ResultsThe overall prevalence of malocclusion among 3‒5-year-old children in Fengxian District of Shanghai was 70.73%, with no statistically significant differences between genders (P>0.05). Deep overbite accounted for the highest proportion (49.66%), followed by Angle Class Ⅱ malocclusion of the second deciduous molars (17.83% on the left, 19.11% on the right), midline deviation (13.23%), anterior crossbite (6.74%), and crowding of the maxillary and mandibular anterior teeth (16.38% for the maxillary dentition, 17.58% for the mandibular dentition). Multivariate analyses showed that habitual mouth breathing during the day (OR=0.509, P=0.009) and morning dry mouth upon waking (OR=1.455, P=0.023) were independent influencing factors for malocclusion. ConclusionThe prevalence of malocclusion is relatively high among preschool children in Fengxian District, Shanghai,with deep overbite being the dominant trait. Sleep-related breathing conditions (such as habitual mouth breathing during the day and morning dry mouth) is an important influencing factor for malocclusion. Public health authorities should strengthen oral health education and promote the correction of bad habits to reduce the prevalence of malocclusion and improve children’s oral health.

Objective:To investigate the relationship between different states of cardiac function and their changes during the course of diabetic foot ulcers(DFU), and to evaluate their impact on patient prognosis.Methods:A retrospective analysis was conducted on 194 DFU patients who were rehospitalized at approximately 3-month intervals. Basic clinical data and cardiac function-related indicators were collected at baseline and follow-up. Patients were followed until death or until November 10, 2024. Outcomes including ulcer healing, recurrence, minor amputation, and death were recorded. Logistic regression models were used to analyze the effects of cardiac function status and its changes on these four outcomes. Results:After treatment, the proportion of patients with NYHA class Ⅱ-Ⅲ decreased significantly from 33.5% at baseline to 21.6%( P=0.009). Serum N-terminal pro-B-type natriuretic peptide(NT-proBNP) level also decreased after treatment compared with baseline [635.85(59.83, 453.28) pg/mL vs 728.67(81.48, 696.15) pg/mL, P=0.055]. Serum NT-proBNP level was significantly higher in the death group compared to the survival group( P=0.002). The proportion of DFU patients with baseline NYHA class Ⅱ-Ⅲ was significantly higher than that in those with class Ⅰ( P=0.012). Regression analysis showed that an improvement in NT-proBNP levels was associated with a lower risk of DFU recurrence( OR=0.378, 95% CI 0.183-0.779, P=0.008), and improvement in NYHA class was associated with a lower mortality risk( OR=0.074, 95% CI 0.020-0.275, P<0.001). Conclusion:Cardiac function status and its changes during the treatment of DFU patients have strong prognostic implications, particularly in predicting the risk of recurrence and death outcomes.

Objective:To explore the intervention effects of the skill training for parents with autism child (STPAC) on toddlers with autism spectrum disorder (ASD).Methods:A multicenter non-randomized concurrent controlled study design was conducted. Thirty children with ASD aged 15-30 months, first diagnosed at the Children′s Hospital of Fudan University, the First Hospital of Jilin University, and Chengdu Women′s and Children′s Central Hospital from 2019 to 2020, were enrolled in the STPAC group. Thirty children with ASD who visited the same hospitals during the same period but refused the STPAC intervention were selected as the control group. The STPAC group received an 8-week intervention (3 h/week) followed by quarterly follow-ups for 1 year, while the control group voluntarily chose community-based routine interventions. The Griffiths development scales-Chinese (GDS-C) was used to assess the developmental levels, and the communication and symbolic behavior scales developmental profile infant-toddler checklist (CSBS-DP-ITC) was completed by the primary caregivers to evaluate social, language and symbolic behavior. The independent samples t-tests or Mann-Whitney U tests, etc.was used for inter-group comparison. The paired t-tests or Wilcoxon signed-rank tests, etc. was used for inter-group pre-post intervention comparison. Results:The STPAC group included 30 children (22 males and 8 females, aged (23.9±2.2) months), and the control group included 30 children (20 males and 10 females, aged (24.2±2.6) months). Before the intervention, there were no statistically differences in GDS-C development quotient (DQ) and CSBS-DP-ITC scores between groups (all P>0.05). After 1-year intervention, GDS-C DQ in personal-social, hearing-language, hand-eye coordination, performance domains of STPAC group and GDS-C DQ in personal-social, hearing-language domains of control group were all increased (all P<0.01). After 1-year intervention, CSBS-DP-ITC scores of both groups were all improved in socia, language, symbolic behavior, and total scores (all P<0.001). GDS-C DQ changes before and after 1 year of intervention in hearing-language, hand-eye coordination, performance domains of the STPAC group were all higher the those of control group (34(15, 48 vs. 10(-4, 39), 11±20 vs. -1±19, 23±25 vs. 8±22, all P<0.05). CSBS-DP-ITC scores changes before and after 1 year of intervention in social and total scores of the STPAC group were both higher the those of control group (10(5, 30) vs. 3(1, 7), 26±17 vs. 11±8, both P<0.001). Conclusion:Compared with the community routine interventions, the STPAC better improves the language, hand-eye coordination, visual-spatial, social communication, and play skills in ASD toddlers.

Objective:To establish tumor specific protein (SP70) targeted tumor cell enrichment technology and to assess applicational value of next-generation sequencing (NGS) analysis for enriched circulating tumor cell (CTC) in precision medicines of non-small cell lung cancer (NSCLC).Methods:The monoclonal antibody NJ001 was covalently coupled to the surface of magnetic beads to build targeted magnetic bead enrichment technology based on SP70. The limit of detection, coincidence rate, interference experiment, recovery test and clinical performance were evaluated. From March 2016 to August 2017, NGS analysis with or without pre-treatment of targeted enrichment for serous fluids of 43 NSCLC in the First Affiliated Hospital with Nanjing Medical University were compared (Kappa or Fisher exact test).Results:The CTC enrichment technology based on SP70 targeted immunomagnetic beads can specifically enrich tumor cells. The limit of detection was 10 4 SPC-A1 cells/L, and the coincidence rate, sensitivity and specificity were 100% (3/3). The endogenous interfering substances such as red blood cells, hemoglobin, white blood cells, epithelial cells and triglycerides had no interfering effects, as well as the exogenous interfering substances such as EDTA-K2, cefoxitin, carboplatin and paclitaxel. The recovery rate was 56.0% (56 000/100 000). A total of 30 gene mutations including 65 loci were found in 43 NSCLC under SP70 targeted enrichment, with a higher detection rate compared with unenrichment method [95.0% (19/20) vs 65.0% (13/20), χ 2=5.625, P=0.044]. Conclusion:In this study, SP70-targeted enriched CTC liquid biopsy method was established, with higher sensitivity and specificity of NGS detection than unenrichment method.

Objective:To evaluate the efficacy of a cetyl alcohol-containing moisturizing cream (referred to as the emollient) in improving clinical symptoms and preventing disease relapse during the remission phase of mild to moderate infantile atopic dermatitis (AD).Methods:A single-center, investigator-blinded, randomized controlled clinical trial was conducted from February 2022 to October 2023. Sixty participants aged >28 days to ≤2 years with mild to moderate AD (Investigator′s Global Assessment (IGA) score 2 or 3) and controlled disease (IGA 0 or 1) were enrolled. Participants were randomized 1∶1 to either the experimental group (topical emollient applied twice daily combined with a moisturizing cleanser for daily bathing) or the control group (moisturizing cleanser alone). Assessments at weeks 2, 4, 8, and 12 included AD relapse rates, clinical severity scores (IGA and Eczema Area and Severity Index (EASI)), and adverse events.Results:Fifty-six participants were included in the full analysis set. The median time to relapse was 12.0 (6.0-72.0) days in the experimental group versus 8.5 (3.0-19.0) days in the control group (log-rank test: χ2=3.079, P=0.213). Relapse rates were 80.77% (21/26) in the experimental group and 95.83% (23/24) in the control group ( χ2=2.682, P=0.101). The experimental group showed significantly lower IGA scores at weeks 2 and 12 ( P<0.05) and lower EASI scores at weeks 4 and 12 ( P<0.05). Adverse event rates were 8.0% (2/25) and 7.7% (2/26) in the experimental and control groups, respectively ( χ2=0.002, P>0.05) Conclusion:The combination of the cetyl alcohol- containing moisturizing cream and moisturizing cleanser during AD remission may improve clinical symptoms and modestly delay relapse, with a low incidence of adverse events.

Objective:To investigate the relationship between different states of cardiac function and their changes during the course of diabetic foot ulcers(DFU), and to evaluate their impact on patient prognosis.Methods:A retrospective analysis was conducted on 194 DFU patients who were rehospitalized at approximately 3-month intervals. Basic clinical data and cardiac function-related indicators were collected at baseline and follow-up. Patients were followed until death or until November 10, 2024. Outcomes including ulcer healing, recurrence, minor amputation, and death were recorded. Logistic regression models were used to analyze the effects of cardiac function status and its changes on these four outcomes. Results:After treatment, the proportion of patients with NYHA class Ⅱ-Ⅲ decreased significantly from 33.5% at baseline to 21.6%( P=0.009). Serum N-terminal pro-B-type natriuretic peptide(NT-proBNP) level also decreased after treatment compared with baseline [635.85(59.83, 453.28) pg/mL vs 728.67(81.48, 696.15) pg/mL, P=0.055]. Serum NT-proBNP level was significantly higher in the death group compared to the survival group( P=0.002). The proportion of DFU patients with baseline NYHA class Ⅱ-Ⅲ was significantly higher than that in those with class Ⅰ( P=0.012). Regression analysis showed that an improvement in NT-proBNP levels was associated with a lower risk of DFU recurrence( OR=0.378, 95% CI 0.183-0.779, P=0.008), and improvement in NYHA class was associated with a lower mortality risk( OR=0.074, 95% CI 0.020-0.275, P<0.001). Conclusion:Cardiac function status and its changes during the treatment of DFU patients have strong prognostic implications, particularly in predicting the risk of recurrence and death outcomes.

Objective:To explore the intervention effects of the skill training for parents with autism child (STPAC) on toddlers with autism spectrum disorder (ASD).Methods:A multicenter non-randomized concurrent controlled study design was conducted. Thirty children with ASD aged 15-30 months, first diagnosed at the Children′s Hospital of Fudan University, the First Hospital of Jilin University, and Chengdu Women′s and Children′s Central Hospital from 2019 to 2020, were enrolled in the STPAC group. Thirty children with ASD who visited the same hospitals during the same period but refused the STPAC intervention were selected as the control group. The STPAC group received an 8-week intervention (3 h/week) followed by quarterly follow-ups for 1 year, while the control group voluntarily chose community-based routine interventions. The Griffiths development scales-Chinese (GDS-C) was used to assess the developmental levels, and the communication and symbolic behavior scales developmental profile infant-toddler checklist (CSBS-DP-ITC) was completed by the primary caregivers to evaluate social, language and symbolic behavior. The independent samples t-tests or Mann-Whitney U tests, etc.was used for inter-group comparison. The paired t-tests or Wilcoxon signed-rank tests, etc. was used for inter-group pre-post intervention comparison. Results:The STPAC group included 30 children (22 males and 8 females, aged (23.9±2.2) months), and the control group included 30 children (20 males and 10 females, aged (24.2±2.6) months). Before the intervention, there were no statistically differences in GDS-C development quotient (DQ) and CSBS-DP-ITC scores between groups (all P>0.05). After 1-year intervention, GDS-C DQ in personal-social, hearing-language, hand-eye coordination, performance domains of STPAC group and GDS-C DQ in personal-social, hearing-language domains of control group were all increased (all P<0.01). After 1-year intervention, CSBS-DP-ITC scores of both groups were all improved in socia, language, symbolic behavior, and total scores (all P<0.001). GDS-C DQ changes before and after 1 year of intervention in hearing-language, hand-eye coordination, performance domains of the STPAC group were all higher the those of control group (34(15, 48 vs. 10(-4, 39), 11±20 vs. -1±19, 23±25 vs. 8±22, all P<0.05). CSBS-DP-ITC scores changes before and after 1 year of intervention in social and total scores of the STPAC group were both higher the those of control group (10(5, 30) vs. 3(1, 7), 26±17 vs. 11±8, both P<0.001). Conclusion:Compared with the community routine interventions, the STPAC better improves the language, hand-eye coordination, visual-spatial, social communication, and play skills in ASD toddlers.

Objective:To establish tumor specific protein (SP70) targeted tumor cell enrichment technology and to assess applicational value of next-generation sequencing (NGS) analysis for enriched circulating tumor cell (CTC) in precision medicines of non-small cell lung cancer (NSCLC).Methods:The monoclonal antibody NJ001 was covalently coupled to the surface of magnetic beads to build targeted magnetic bead enrichment technology based on SP70. The limit of detection, coincidence rate, interference experiment, recovery test and clinical performance were evaluated. From March 2016 to August 2017, NGS analysis with or without pre-treatment of targeted enrichment for serous fluids of 43 NSCLC in the First Affiliated Hospital with Nanjing Medical University were compared (Kappa or Fisher exact test).Results:The CTC enrichment technology based on SP70 targeted immunomagnetic beads can specifically enrich tumor cells. The limit of detection was 10 4 SPC-A1 cells/L, and the coincidence rate, sensitivity and specificity were 100% (3/3). The endogenous interfering substances such as red blood cells, hemoglobin, white blood cells, epithelial cells and triglycerides had no interfering effects, as well as the exogenous interfering substances such as EDTA-K2, cefoxitin, carboplatin and paclitaxel. The recovery rate was 56.0% (56 000/100 000). A total of 30 gene mutations including 65 loci were found in 43 NSCLC under SP70 targeted enrichment, with a higher detection rate compared with unenrichment method [95.0% (19/20) vs 65.0% (13/20), χ 2=5.625, P=0.044]. Conclusion:In this study, SP70-targeted enriched CTC liquid biopsy method was established, with higher sensitivity and specificity of NGS detection than unenrichment method.

Objective:To evaluate the efficacy of a cetyl alcohol-containing moisturizing cream (referred to as the emollient) in improving clinical symptoms and preventing disease relapse during the remission phase of mild to moderate infantile atopic dermatitis (AD).Methods:A single-center, investigator-blinded, randomized controlled clinical trial was conducted from February 2022 to October 2023. Sixty participants aged >28 days to ≤2 years with mild to moderate AD (Investigator′s Global Assessment (IGA) score 2 or 3) and controlled disease (IGA 0 or 1) were enrolled. Participants were randomized 1∶1 to either the experimental group (topical emollient applied twice daily combined with a moisturizing cleanser for daily bathing) or the control group (moisturizing cleanser alone). Assessments at weeks 2, 4, 8, and 12 included AD relapse rates, clinical severity scores (IGA and Eczema Area and Severity Index (EASI)), and adverse events.Results:Fifty-six participants were included in the full analysis set. The median time to relapse was 12.0 (6.0-72.0) days in the experimental group versus 8.5 (3.0-19.0) days in the control group (log-rank test: χ2=3.079, P=0.213). Relapse rates were 80.77% (21/26) in the experimental group and 95.83% (23/24) in the control group ( χ2=2.682, P=0.101). The experimental group showed significantly lower IGA scores at weeks 2 and 12 ( P<0.05) and lower EASI scores at weeks 4 and 12 ( P<0.05). Adverse event rates were 8.0% (2/25) and 7.7% (2/26) in the experimental and control groups, respectively ( χ2=0.002, P>0.05) Conclusion:The combination of the cetyl alcohol- containing moisturizing cream and moisturizing cleanser during AD remission may improve clinical symptoms and modestly delay relapse, with a low incidence of adverse events.