BackgroundAttention deficit hyperactivity disorder （ADHD） is a neurodevelopmental disorder characterized by age-inappropriate inattention， excessive activities incongruous with setting， and emotional impulsivity. Subthreshold ADHD （sADHD） is clinically defined as the presence of ADHD symptoms that do not meet the full diagnostic criteria for ADHD. Children with sADHD exhibit deficits in executive function， demonstrate more conduct， learning， and anxiety-related problems compared to typically developing children， and show even poorer working memory performance than children diagnosed with ADHD. Currently， there is limited epidemiological research on sADHD in China， with few studies simultaneously investigating the prevalence of both ADHD and sADHD in children. ObjectiveTo investigate the prevalence of ADHD and sADHD among children aged 6–13 years in Chongqing， analyzing their distribution characteristics within this population， with the aim of providing references for developing preventive measures against both ADHD and sADHD. MethodsFrom October to November 2023， a total of 3 398 students in grades 1–6 from six primary schools in Jiangbei District， Chongqing were selected using a stratified cluster random sampling method. The occurrence of ADHD and sADHD was evaluated by using the short version （18-item version） of the Swanson， Nolan， and Pelham IV rating scales （SNAP-IV） and the Chinese vision of Schedule for Affective Disorder and Schizophrenia for School-aged Children-Present and Lifetime Version （K-SADS-PL）. ResultsThe ADHD detection rate among children in Chongqing was 1.90% （95% CI： 0.014–0.024）. Boys showed a significantly higher ADHD detection rate than girls （χ²=7.733， P=0.005）. No statistically significant differences were found in ADHD detection rates across different grades or age groups （χ²=7.347， 12.362， P>0.05）. The sADHD detection rate was 6.32% （95% CI： 0.054–0.072）. Similarly， boys exhibited significantly higher sADHD detection rates than girls （χ²=21.005， P<0.01）. Significant differences emerged across different grades （χ²=20.559， P=0.001）， while no statistically significant difference was observed in age groups （χ²=12.070， P=0.060）. ConclusionThe ADHD detection rates were comparable across all grade levels and age groups from 6–13 years old. Second-grade children demonstrated notably higher sADHD rates compared to other grades， while boys demonstrated higher prevalence rates than girls for both ADHD and sADHD. ［Funded by Science and Health Joint Medical Research Project in Jiangbei District， Chongqing City in the Second Half of 2023 （number， 2023JBKWLH022）］

Objective To investigate the regulatory effects and molecular mechanisms of extracellular vesicles(EVs)secreted by adri-amycin(ADR)-resistant breast cancer cells on vascular endothelial cells related to tumor microenvironment.Methods The ADR-sen-sitive breast cancer cell line(MCF-7)and its ADR-resistant counterpart(MCF-7-ADR)were established in vitro.Then,the EVs were isolated from the supernatant of cell culture using a commercial kit.The fluorescence labeling and tracking technology using PKH67 was used to validate the internalization of EVs by human umbilical vein endothelial cell(HUVEC).The effects of EVs on the proliferation,migration,and angiogenesis of HUVEC were assessed by the colony formation,scratch wound healing,and tube formation assays,re-spectively.The expression level of CD31 protein was detected by Western blot,and the levels of vascular endothelial growth factor(VEGF)in cell supernatant were determined by ELISA.The microRNA(miRNA)sequencing technology was utilized to identify the differentially expressed miRNAs in EVs secreted by ADR-resistant cells.Results Compared with the ADR-sensitive strain,the EVs secreted by the ADR-resistant strain significantly enhanced the proliferation(colony number increased by 38.7%,P=0.011),migra-tion(scratch healing rate improved by 42.3%,P=0.004),and tube formation ability(tube formation rates increased by 1.8 times and 2.3 times at 4 h and 16 h,respectively,P=0.044 and 0.010)of HUVEC.Western blot analysis showed that the expression level of CD31 in the EVs secreted by the ADR-resistant strain increased by 1.85 times compared with the ADR-sensitive strain(P=0.039).The results of ELISA showed that the expression level of VEGF increased by 2.12 times(P=0.012).The miRNA sequencing identified a differentially expressed miRNA profile.Conclusion The ADR-resistant breast cancer cells significantly enhance the proliferation,migration,and angiogenesis of vascular endothelial cells by secreting EVs rich in specific miRNA,and its mechanism may be related to the up-regulation of CD31/VEGF protein levels mediated by EVs,suggesting that the drug-resistant cells reshaping the tumor vascular microenvironment through EVs may become a new therapeutic target of chemotherapy resistance in breast cancer.

Objective To investigate the regulatory effects and molecular mechanisms of extracellular vesicles(EVs)secreted by adri-amycin(ADR)-resistant breast cancer cells on vascular endothelial cells related to tumor microenvironment.Methods The ADR-sen-sitive breast cancer cell line(MCF-7)and its ADR-resistant counterpart(MCF-7-ADR)were established in vitro.Then,the EVs were isolated from the supernatant of cell culture using a commercial kit.The fluorescence labeling and tracking technology using PKH67 was used to validate the internalization of EVs by human umbilical vein endothelial cell(HUVEC).The effects of EVs on the proliferation,migration,and angiogenesis of HUVEC were assessed by the colony formation,scratch wound healing,and tube formation assays,re-spectively.The expression level of CD31 protein was detected by Western blot,and the levels of vascular endothelial growth factor(VEGF)in cell supernatant were determined by ELISA.The microRNA(miRNA)sequencing technology was utilized to identify the differentially expressed miRNAs in EVs secreted by ADR-resistant cells.Results Compared with the ADR-sensitive strain,the EVs secreted by the ADR-resistant strain significantly enhanced the proliferation(colony number increased by 38.7%,P=0.011),migra-tion(scratch healing rate improved by 42.3%,P=0.004),and tube formation ability(tube formation rates increased by 1.8 times and 2.3 times at 4 h and 16 h,respectively,P=0.044 and 0.010)of HUVEC.Western blot analysis showed that the expression level of CD31 in the EVs secreted by the ADR-resistant strain increased by 1.85 times compared with the ADR-sensitive strain(P=0.039).The results of ELISA showed that the expression level of VEGF increased by 2.12 times(P=0.012).The miRNA sequencing identified a differentially expressed miRNA profile.Conclusion The ADR-resistant breast cancer cells significantly enhance the proliferation,migration,and angiogenesis of vascular endothelial cells by secreting EVs rich in specific miRNA,and its mechanism may be related to the up-regulation of CD31/VEGF protein levels mediated by EVs,suggesting that the drug-resistant cells reshaping the tumor vascular microenvironment through EVs may become a new therapeutic target of chemotherapy resistance in breast cancer.

Objective:To investigate the clinicopathological and molecular genetic characteristics of Crohn′s disease (CD).Methods:A retrospective analysis was conducted on 52 CD patients who underwent surgical resection at the First Affiliated Hospital of Nanjing Medical University between January 2014 and June 2023. Clinical presentations and histopathological features were assessed. Whole-genome sequencing was performed on 17 of the samples, followed by sequencing and pathway enrichment analyses. Immunohistochemistry was used to assess the expression of frequently mutated genes.Results:Among the 52 patients, 34 were males and 18 were females, male-to-female ratio was 1.9∶1.0, with a median age of 45 years at surgery and 35 years at diagnosis. According to the Montreal classification, A3 (51.9%,27/52), B2 (61.5%, 32/52), and L3 (50.0%,26/52) subtypes were the most predominant. Abdominal pain and diarrhea were the common symptoms. Histopathological features seen in all 52 patients included transmural inflammation, disruption of cryptal architecture, lymphoplasmacytic infiltration, varying degrees of submucosal fibrosis and thickening, increased enteric nerve fibers and neuronal proliferation. Mucosal defects, fissure ulcers, abscesses, pseudopolyps, and adenomatous proliferation were also observed in 51 (98.1%), 38 (73.1%), 28 (53.8%), 45 (86.5%), and 28 (53.8%) cases, respectively. Thirty-one (59.6%) cases had non-caseating granulomas, and 3 (5.8%) cases had intestinal mucosal glandular epithelial dysplasia. Molecular analysis showed that 12/17 CD patients exhibited mutations in at least one mucin family gene (MUC2, MUC3A, MUC4, MUC6, MUC12, MUC17), and MUC4 was the most frequently mutated in 7/17 of cases. Immunohistochemical stains showed reduced MUC4 expression in epithelial cells, with increased MUC4 expression in the epithelial surface, particularly around areas of inflammatory cell aggregation; and minimal expression in the lower half of the epithelium.Conclusions:CD exhibits diverse clinical and pathological features, necessitating a comprehensive multidimensional analysis for diagnosis. Mutations and expression alterations in mucin family genes, particularly MUC4, may play crucial roles in the pathogenesis of CD.

Objective To explore the potential value of coagulation indicators and their combinations for metastasis and postoperative recurrence in renal cell carcinoma(RCC)patients.Methods A total of 39 RCC patients with metastasis and postoperative recurrence were enrolled,and the controls groups included 64 RCC patients without metastasis,15 patients with renal vascular leiomyoma/oncocy-toma and 118 patients with renal cysts.All the patients were diagnosed and hospitalized in the Department of Urology of Nanjing Hospi-tal of Chinese Medicine affiliated with Nanjing University of Chinese Medicine and Jinling Hospital affiliated with Nanjing University School of Medicine from January 2018 to June 2024.The coagulation indicators in the blood samples were measured.The levels of plas-ma prothrombin time(PT),activated partial thromboplastin time(APTT),thrombin time(TT)and fibrinogen(Fib)were analyzed by clotting method.The levels of fibrinogen degradation products(FDP)and D-Dimer(DD)were analyzed by immunological turbidim-etry assay,and the levels of antithrombin Ⅲ(AT3)were analyzed by chromogenic substrate method,respectively.The differences of coagulation indicators between RCC patients with metastasis and postoperative recurrence and the controls were assessed by Mann-Whit-ney U test.The potential value of coagulation indicators and their combinations for predicating metastasis and postoperative recurrence in RCC patients was evaluated through Logistic regression and ROC curve analysis.Results The plasma levels of the seven coagulation indicators showed significant differences between the patients with RCC metastatic/recurrent and the controls(all with P＜0.05).The results of ROC curve analysis showed that the plasma levels of all the 7 coagulation index could distinguish the metastatic/recurrent RCC patients from the controls,and the DD level showed the highest value with the AUCROC=0.913(95％CI:0.859 to 0.967).Multi-ple logistic regression analysis revealed that the plasma levels of Fib,FDP,DD,and AT3 were associated with RCC metastasis/recur-rence.Logistic regression analysis combined with ROC curve analysis revealed that the panel of Fib,FDP,DD and AT3 had the best performance for discriminating RCC metastasis/recurrence along with the AUCROC of 0.920(95％CI:0.865 to 0.974).Conclusion Alterations of the levels of plasma Fib,DD,FDP and AT3 may be closely related to postoperative recurrence and metastasis in RCC patients,and the panel of Fib,FDP,DD and AT3 has the potential to serve as the monitoring and evaluation indicators for RCC recur-rence and metastasis.

Objective:To detect the changes in the levels of miR-21-5p and miR-125a-5p modified with 3′-terminal 2′-O-methylation (3′t-2′Ome) in serum extracellular vesicles (EV) of non-small cell lung cancer (NSCLC) patients, and evaluate their value as auxiliary screening molecular markers for NSCLC patients.Method:A retrospective analysis was conducted on the data of 69 NSCLC patients diagnosed at the Eastern Theater Command General Hospital from May 1st to October 31st,2023, as well as 65 age and gender matched healthy controls during the same period. Two real-time fluorescence quantitative PCR (RT-qPCR) techniques, namely stem-loop method and poly (A) tailed method, were used to detect the levels of 3′t-2′Ome-miR-21-5p and 3′t-2′-Ome miR-125a-5p in serum EV of NSCLC patients and controls.Analyze the correlation between the levels of two types of 3′t-2 ′Ome miRNAs and the differences in clinical stage, pathological classification, and other tumor indicators in patients. Receiver operating characteristic (receiveroperating curve, ROC) curves were used to analyze the efficacy of 3′t-2 ′Ome miR-21-5p and 3′t-2′ Ome miR-125a-5p in serum EV, as well as their combination, in diagnosing NSCLC.Result:Compared with the control group, the levels of 3′t-2′Ome-miR-21-5p in serum EV of NSCLC patients increased [(0.30±0.05) vs (0.35±0.09), t=3.32, P=0.001], while the levels of 3′t-2′Ome-miR-125a-5p decreased [(0.33±0.06 vs 0.25±0.06, t=7.45, P<0.001]. The differences were statistically significant. The expression levels of 2′Ome-miR-21-5p in EV were also significantly elevated in the NSCLC patients at 0-Ⅱ stage, adenocarcinoma patients, and squamous cell carcinoma patients, respectively. Notably, the levels of EV 3′t-2′Ome-miR-21-5p was also statistically significant between the adenocarcinoma patients and squamous cell carcinoma patients [(0.34±0.85) vs (0.40±0.12), P<0.05]. ROC curve analysis showed that the levels of 3′t-2 ′Ome miR-21-5p and 3′t-2′ Ome miR-125a-5p in serum EV, as well as their combined AUC for discriminating NSCLC patients, were 0.647(95% CI 0.550-0.743), 0.825(95% CI 0.756-0.894) and 0.860(95% CI 0.797-0.923), respectively. The sensitivity was 92.3%, 80.0%, 89.2%, and the specificity was 46.4%, 73.9%, and 78.3%, respectively. Conclusion:There are changes in the levels of 2 ′Ome miR-21-5p and 2′ Ome miR-125a-5p in the serum EV of NSCLC patients, and the combined detaction has the potential as an auxiliary screening molecular marker of NSCLC patients.

Background Type 2 diabetes mellitus is one of the most prevalent diseases,which imposes a heavy burden on patients' families and the society.Sleep disorders are recognized as risk factors for the development of diabetes,which may affect the onset and development of diabetes through neuro-endocrino-metabolic pathways,so identifying the factors responsible for the sleep quality of diabetic patients is of great importance in improving their sleep quality.Objective To investigate the relationship among fear of disease progression,executive function and sleep quality in patients with type 2 diabetes mellitus,so as to provide references for improvement of sleep quality in patients with type 2 diabetes mellitus.Methods A sample of 197 patients with type 2 diabetes mellitus who were admitted to the Endocrinology Department of the Second Affiliated Hospital of the Air Force Military Medical University from January to May 2023 and met the criteria defined in the Guideline for the Prevention and Treatment of Type 2 Diabetes Mellitus in China(2020 edition)were consecutively selected.All subjects were assessed using Fear of Progression Questionnaire-Short Form(FoP-Q-SF),Behavior Rating Inventory of Executive Function-Adult version(BRIEF-A)and Pittsburgh Sleep Quality Index(PSQI).Then the Process macro for SPSS(Model 4)and Bootstrap technique were applied to examine the mediating effect of executive function on the relationship between fear of disease progression and sleep quality in patients with type 2 diabetes mellitus.Results ①75 patients(38.07%)with type 2 diabetes mellitus were found to have sleep problems.②PSQI score in patients with type 2 diabetes mellitus was positively correlated with FoP-Q-SF score and BRIEF-A score(r=0.159,0.287,P<0.01).③Executive function mediated the relationship between fear of disease progression and sleep quality,the indirect value was 0.076(95%CI:0.022～0.146),accounting for 39.58%of the total effect.Conclusion Sleep disorders are common in patients with type 2 diabetes mellitus,and executive function may play a medicating role in the relationship between fear of disease progression and sleep quality.

Objective To explore the potential value of coagulation indicators and their combinations for metastasis and postoperative recurrence in renal cell carcinoma(RCC)patients.Methods A total of 39 RCC patients with metastasis and postoperative recurrence were enrolled,and the controls groups included 64 RCC patients without metastasis,15 patients with renal vascular leiomyoma/oncocy-toma and 118 patients with renal cysts.All the patients were diagnosed and hospitalized in the Department of Urology of Nanjing Hospi-tal of Chinese Medicine affiliated with Nanjing University of Chinese Medicine and Jinling Hospital affiliated with Nanjing University School of Medicine from January 2018 to June 2024.The coagulation indicators in the blood samples were measured.The levels of plas-ma prothrombin time(PT),activated partial thromboplastin time(APTT),thrombin time(TT)and fibrinogen(Fib)were analyzed by clotting method.The levels of fibrinogen degradation products(FDP)and D-Dimer(DD)were analyzed by immunological turbidim-etry assay,and the levels of antithrombin Ⅲ(AT3)were analyzed by chromogenic substrate method,respectively.The differences of coagulation indicators between RCC patients with metastasis and postoperative recurrence and the controls were assessed by Mann-Whit-ney U test.The potential value of coagulation indicators and their combinations for predicating metastasis and postoperative recurrence in RCC patients was evaluated through Logistic regression and ROC curve analysis.Results The plasma levels of the seven coagulation indicators showed significant differences between the patients with RCC metastatic/recurrent and the controls(all with P＜0.05).The results of ROC curve analysis showed that the plasma levels of all the 7 coagulation index could distinguish the metastatic/recurrent RCC patients from the controls,and the DD level showed the highest value with the AUCROC=0.913(95％CI:0.859 to 0.967).Multi-ple logistic regression analysis revealed that the plasma levels of Fib,FDP,DD,and AT3 were associated with RCC metastasis/recur-rence.Logistic regression analysis combined with ROC curve analysis revealed that the panel of Fib,FDP,DD and AT3 had the best performance for discriminating RCC metastasis/recurrence along with the AUCROC of 0.920(95％CI:0.865 to 0.974).Conclusion Alterations of the levels of plasma Fib,DD,FDP and AT3 may be closely related to postoperative recurrence and metastasis in RCC patients,and the panel of Fib,FDP,DD and AT3 has the potential to serve as the monitoring and evaluation indicators for RCC recur-rence and metastasis.

Objective To examine the expression pattern of miR-135a-5p and miR-135b-5p in seminal plasma as well as the clinical value of their variation in the male patients with infertility.Methods The clinical samples of 5 kinds,including serum,morning urine,pleural effusion,cerebrospinal fluid and seminal plasma,were collected from 30 patients respectively in the Department of Clini-cal Laboratory,Jiangsu Province Hospital of Chinese Medicine from January 2019 to December 2021.In addition,the frozen seminal plasma samples from the patients with non-obstructive azoospermia,asthenozoospermia of 54 cases for each illness statuses,and 54 healthy controls with fertile ability from 2010 to 2019 in Jiangsu Province Hospital of Chinese Medicine and Jinling Hospital were also selected.Fluorescent quantitative RT-qPCR analyses were applied to measure the expression levels of miR-135a-5p and miR-135b-5p in seminal plasma.Receiver operating characteristic curve(ROC)was used to construct the area under the curve(AUCROC)to evalu-ate the ability for discriminating infertility males from healthy controls.Correlation analyses were performed to explore the correlations between the levels of miR-135a-5p and miR-135b-5p in seminal plasma and other semen parameters.Results The concentrations of miR-135a-5p and miR-135b-5p in the samples from five different kinds of body fluid showed significantly difference(H=64.88,P＜0.01 and H=64.7,P＜0.01,respectively).The highest levels were found in serum and seminal plasma,and the lowest levels were in u-rine.The concentrations of miR-135a-5p and miR-135b-5p in seminal plasma were also showed marked difference among the azoosper-mia group,asthenozoospermia group and healthy fertile control group(H=32.14,P＜0.01 and H=21.37,P＜0.01,respectively).Compared to the fertile control group,the levels of miR-135a-5p and miR-135b-5p in seminal plasma were significantly downregulated in azoospermia patients(U=687,P＜0.01 and U=843,P＜0.01,respectively).The contents of the two miRNAs showed no statistical-ly difference between asthenozoospermia patients and healthy fertile controls,but showed markedly difference between the two patients groups(U=635,P＜0.01 and U=779,P＜0.01,respectively).Receiver operating characteristic curve(ROC)analyses results re-vealed that levels of miR-135a-5p and miR-135b-5p in seminal plasma could successfully discriminate the azoospermia patients from fertile controls and asthenozoospermia patients with the AUCROC of 0.764(95％CI:0.675 to 0.854),0.711(95％CI:0.612 to 0.810)and 0.782(95％CI:0.695 to 0.869),and 0.733(95％CI:0.637 to 0.829),respectively.Correlation analyses demonstrated that the concentrations of miR-135a-5p and miR-135b-5p in seminal plasma were significantly associated with sperm concentration,total sperm motility,and percentages of progressive sperm and non-progressive sperm(all with the P-value＜0.01).Conclusion miR-135a-5p and miR-135b-5p are the highly expressed miRNA in seminal plasma,and significantly decreased in azoospermia patients.Both of them may play pivot roles in male infertility.

Objective:To observe the effect of supplementing vacuum sealing drainage with hyperbaric oxygen in the short term treatment of diabetic foot ulcers.Methods:A total of 156 persons diagnosed with diabetic foot ulcers were randomly divided into a control group and a treatment group, each of 78. Both groups received life guidance and active treatment to lower blood sugar and lipids, as well as anti-infection treatment guided by bacterial cultures. Both groups′ wounds were debrided. The wound was then covered with foam, sealed, and negative pressure of -75 to -100mmHg was applied during 1 week of drainage. Two courses of this treatment were applied. In addition, the treatment group received hyperbaric oxygen daily during the two weeks. The exposure pressure was incrased to 0.25MPa over 15min with 100% oxygen. That was inhaled in two 30min sessions with a 10min interval. The pressure then decompressed at a constant rate for 25 minutes. Wound healing, hemorheology, wound granulation tissue staining and any changes in TGF-β1 were observed before as well as after 7 and 14 days of the treatment.Results:The average wound size and symptom score of both groups had improved significantly after the treatment, with the largest effect in the treatment group during the first week. Both groups′ hemorheology had improved significantly after one week, but the treatment group′s improvement was greater. After 2 weeks, however, there was no significant difference in the average hemorheologic indicators for either group compared with before the treatment. Hematoxylin-eosin staining of the wound tissues showed that there were many inflamed cells before the treatment, with relatively little fresh granulation tissue or new blood vessels. After one week of treatment much new granulation tissue was observed under the microscope in both groups, with no significant difference between them. One week later, there was still much granulation tissue in the control group, but slightly less in the treatment group. The ave-rage post-treatment TGF-β1 protein levels in the wound tissues of both groups were significantly higher than before the treatment, but after two weeks the average TGF-β1 protein level had decreased significantly in the treatment group compared with the control group.Conclusions:One week of hyperbaric oxygen treatment can effectively improve the hemorheology of persons with diabetic foot ulcers, promote the proliferation of granulation tissue and fibroblasts, and increase the level of TGF-β1 protein in the wound tissues. However, the effects of hyperbaric oxygen treatment weaken gradually with time.