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 effect and mechanisms of celecoxib on right heart function in mice with acute high-altitude hypoxia exposure.Methods Male C57BL/6J mice(7 weeks old)were housed in a hypobaric chamber simulating an altitude of 5 800 m for 2 d to establish an animal model of acute hypobaric hypoxia.①Eighteen mice were randomly assigned to plain+saline(P+S),high-altitude hypoxia exposure+saline(H+S),and high-altitude hypoxia exposure+celecoxib(H+Cel).Body weight and routine blood indicators were measured,and cardiac ultrasound examination were performed for heart rate(HR),pulmonary artery acceleration time to ejection time ratio(AT/ET),tricuspid annular plane systolic excursion(TAPSE),tricuspid annular systolic velocity(S'),and left ventricular ejection fraction(LVEF)and fractional shortening(FS).Targeted metabolomic profiling was applied to detect the cardiac arachidonic acid(AA)metabolite levels.The contents of 12,13-dihydroxy-9Z-octadecenoic acid(12,13-diHOME)in the heart,liver,brown adipose tissue,and plasma were quantified by ELISA.② Eighteen mice were randomly assigned into plain+saline(P+S),high-altitude hypoxia exposure+saline(H+S)and high-altitude hypoxia exposure+12,13-diHOME(H+di)groups.Body weight,routine blood tests,and echocardiography were performed as above.③ Thirty-two mice were randomly divided into high-altitude hypoxia exposure+saline(H+S),high-altitude hypoxia exposure+celecoxib(H+Cel),high-altitude hypoxia exposure+soluble epoxide hydrolase inhibitor(sEHI)(H+sEHI),and high-altitude hypoxia exposure+sEHI+celecoxib(H+sEHI+Cel)groups.Body weight,routine blood tests,and echocardiography were performed as above.Cardiac and plasma contents of 12,13-diHOME and epoxyeicosatrienoic acids(EETs)were measured by ELISA.Results ① Compared to the P+S group,the H+S group exhibited significantly reduction of cardiac 12,13-diHOME level(P<0.001),increased counts of white blood cells(WBC)and neutrophils(P<0.01)and decreased TAPSE,S'and AT/ET both at resting state and under stress(P<0.01,P<0.001).Compared to the H+S group,the H+Cel group exhibited significantly increase of cardiac 12,13-diHOME level(P<0.05),reduced WBC and lymphocyte counts(P<0.01,P<0.05)and improved TAPSE and S'levels at resting state and under stress(P<0.01,P<0.001).② Compared to the H+S group,the H+di group demonstrated significantly improvement of TAPSE at basal and under stress(P<0.001)and a trend towards improved TAPSE at resting state(P=0.0532),but no obvious differences was observed in WBC and neutrophil counts between the H+di group and the H+S group.③ Compared to the H+Cel group,both the H+sEHI and H+sEHI+Cel groups exhibited significantly reduction of cardiac 12,13-diHOME level(P<0.01,P<0.05)though no statistical changes in cardiac function indicators.Compared to the H+S group,WBC counts and lymphocyte were decreased,and serum EETs level was incrased in the H+Cel group,H+sEHI group and H+sEHI+Cel group(P<0.01,P<0.001).Conclusion Celecoxib can elevate cardiac level of 12,13-diHOME and improves right heart function in mice after acute high-altitude hypoxia exposure through the CYP450-sEH metabolic pathway.

Objective To assess stromal fibrosis in epidermal growth factor receptor(EGFR)mutant lung adenocarcinoma and its association with resistance to targeted therapy and patient prognosis.Methods Medical records of 207 patients diagnosed with EGFR-mutant advanced lung adenocarcinoma who received treatment at a hospital between January 2021 and December 2022 were reviewed.A total of 86 patients were ultimately included based on their prognosis and survival duration.These patients were categorized into a resistance group(32 cases)and a non-resistance group(54 cases),depending on whether they developed resistance to targeted therapy within one year.Additionally,patients were classified into mild,moderate,and severe fibrosis groups according to the extent of fibrosis observed.Clinical and pathological characteristics,as well as fibrosis levels,were compared between the two groups.Factors influencing the development of resistance to targeted therapy in patients with EGFR-mutant lung adenocarcinoma were analyzed,and the survival outcomes of patients with varying degrees of fibrosis were evaluated during follow-up.Results In the resistance group,the prevalence of EGFR exon 20 insertion mutations,elevated CA125 levels,and the presence of moderate-to-severe fibrosis were significantly higher compared to the non-resistance group(P<0.05).Multivariate logistic regression analysis revealed that EGFR exon 20 inser-tion mutation(OR=3.691,95%CI:1.043～13.057),elevated CA125 levels(OR=4.104,95%CI:1.160～14.517),and moderate-to-severe fibrosis(OR=3.959,95%CI:1.410～11.115)were independent risk factors associated with resistance to targeted therapy among patients with EGFR-mutant lung adenocarcinoma(P<0.05).The Cox proportional hazards model demonstrated a C-index of 0.72(95%CI:0.65～0.79),with area under the curve(AUC)values for 1-year and 2-year survival predictions of 0.781 and 0.734,respectively.EGFR exon 20 insertion mutation(HR=3.691),moderate-to-severe fibrosis(HR=3.959),and elevated CA125 levels(HR=4.104)were identified as independent prognostic factors for overall survival in these patients following targeted therapy.The median progression-free survival(PFS)for patients with mild,moderate,and severe fibrosis was 10.5 months,7.2 months,and 3.9 months,respectively,while the median overall survival(OS)was 21.4 months,16.1 months,and 11.5 months,respectively.Statistically significant differences in both PFS and OS were observed across the three fibrosis severity groups.(P<0.05).Conclusion The extent of stromal fibrosis in EGFR-mutant lung adenocarcinoma influences resistance to targeted therapy,and the progression of fibrosis is correlated with an unfavorable prognosis.

Objective:To construct a sizeable naive human Fab phage display antibody library to screen high-affinity specific antibodies in vitro. Methods:Total RNA was extracted from peripheral blood mononuclear cells (PBMCs) of 126 healthy individuals, subsequently reverse-transcribed into cDNA, and used as a template. PCR amplification was performed to obtain the V H from IgG, IgM and light chain κ, λ, separately, with the initial PCR products serving as templates for a second round of PCR. Overlap extension PCR was employed to generate fragments of the κ and λ light chains. These fragments were ligated with the phage vector pNC3, which harbors the variable region 1 of the heavy chain, to construct a recombinant phage plasmid. This plasmid was then electroporated into competent Escherichia Coli TG1 cells to establish a naive human Fab phage display antibody library. One hundred clones were randomly selected for identification and sequencing, and antibody gene polymorphisms were analyzed using the IMGT database and MAFFT software. Recombinant α-hemolysin from Staphylococcus aureus was utilized to screen Fab antibody fragments through biopanning of the antibody library, followed by random selection of phage ELISA-identified clones. The positive clones (antigen A450∶blank control A450≥2.1) were sequenced. Results:Two large naive Fab phage display antibody libraries were successfully constructed, in which the capacity of κ and λ chain antibody libraries were 1.25×10 11 and 1.54×10 11, respectively. The titers for two antibody libraries were 6.04×10 13 CFU/ml and 3.50×10 13 CFU/ml. The positive transformation insertion rates for κ and λ chain antibody libraries were 96% (96/100) and 100% (100/100), respectively. Sequence analysis revealed that all antibody sequences were unique. The amino acid sequences in the skeletal region were relatively conserved. In contrast, significant variations in the length of the complementarity determining region (CDR) were found, and the diversity of amino acid sequence of the complementary determining region was high, especially the CDR3. Analysis using the IMGT database indicated that the sequences exhibited a broad distribution across variable-diversity-joining gene families. After six rounds of panning, specific phage antibodies enrichment targeting α-hemolysin were achieved. A total of 142 monoclonal antibodies were sequenced, yielding 8 distinct Fab antibody sequences. Conclusion:This study successfully constructed two naive human Fab phage display antibody libraries with large capacity and good diversity, which can be used for screening human antibodies for serum epidemiology.

This paper focuses on the application of health big data in cancer screening. Firstly, the sources and characteristics of health big data are introduced, then the commonly used epidemiological designs and analytical techniques in hospital-based cancer screening studies are summarized and the application scenarios of such studies are described. Finally, the challenges and future development in the application of health big data are analyzed to provide reference for the future studies.

Objective:To investigate the risk factors for severe postpartum hemorrhage (SPPH) during vaginal delivery of twin pregnancy.Methods:A retrospective analysis was conducted on clinical data from twin pregnancies ≥28 weeks′ gestation undergoing vaginal delivery at Peking University Third Hospital between January 2016 and December 2023. The twin pregnant women were divided into the SPPH group (postpartum hemorrhage ≥1 000 ml within 24 hours) with 22 cases and the non-SPPH group with 171 cases. The differences between the two groups were compared and the risk factors for SPPH were analyzed.Results:(1) The incidence of SPPH during vaginal delivery in twin pregnancies was 11.4% (22/193). The causes of SPPH included 12 cases (54.5%, 12/22) of simple uterine atony, 4 cases (18.2%, 4/22) of uterine atony combined with vaginal lacerations after forceps delivery, and 6 cases (27.3%, 6/22) of uterine atony combined with placental factors. (2) The age and postpartum hospital stay in the SPPH group were significantly higher than those in the non-SPPH group (all P<0.05). Compared to the non-SPPH group, the proportion of hypertensive disorders in pregnancy, accreta placenta implantation, and anemia in the SPPH group were significantly increased, and the birth weight of newborn 1st, the sum of the birth weights of two newborns, the duration of the second stage of labor, and the proportion of labor followed induction were also significantly increased (all P<0.05). (3) Multivariate analysis showed that age ≥38 years ( OR=16.785, 95% CI: 2.679-105.166; P=0.003), the second stage of labor ≥90 minutes ( OR=9.670, 95% CI: 2.532-36.930; P=0.001), hypertensive disorders in pregnancy ( OR=5.945, 95% CI: 1.702-20.761; P=0.005), and anemia ( OR=8.048, 95% CI: 2.086-31.049; P=0.002) were independent risk factors for SPPH in twin pregnancies during vaginal delivery. Conclusions:Anemia should be actively corrected during twin pregnancy. For twin pregnant women with advanced age, hypertensive disorders in pregnancy, or other risk factors of SPPH, if vaginal delivery is chosen, attention should be paid to the management of labor duration, dynamic assessment of the risk of postpartum hemorrhage, and proactive measures should be taken to ensure a smooth vaginal delivery and effectively reduce the incidence of SPPH.

Objective:To investigate the risk factors for severe postpartum hemorrhage (SPPH) during vaginal delivery of twin pregnancy.Methods:A retrospective analysis was conducted on clinical data from twin pregnancies ≥28 weeks′ gestation undergoing vaginal delivery at Peking University Third Hospital between January 2016 and December 2023. The twin pregnant women were divided into the SPPH group (postpartum hemorrhage ≥1 000 ml within 24 hours) with 22 cases and the non-SPPH group with 171 cases. The differences between the two groups were compared and the risk factors for SPPH were analyzed.Results:(1) The incidence of SPPH during vaginal delivery in twin pregnancies was 11.4% (22/193). The causes of SPPH included 12 cases (54.5%, 12/22) of simple uterine atony, 4 cases (18.2%, 4/22) of uterine atony combined with vaginal lacerations after forceps delivery, and 6 cases (27.3%, 6/22) of uterine atony combined with placental factors. (2) The age and postpartum hospital stay in the SPPH group were significantly higher than those in the non-SPPH group (all P<0.05). Compared to the non-SPPH group, the proportion of hypertensive disorders in pregnancy, accreta placenta implantation, and anemia in the SPPH group were significantly increased, and the birth weight of newborn 1st, the sum of the birth weights of two newborns, the duration of the second stage of labor, and the proportion of labor followed induction were also significantly increased (all P<0.05). (3) Multivariate analysis showed that age ≥38 years ( OR=16.785, 95% CI: 2.679-105.166; P=0.003), the second stage of labor ≥90 minutes ( OR=9.670, 95% CI: 2.532-36.930; P=0.001), hypertensive disorders in pregnancy ( OR=5.945, 95% CI: 1.702-20.761; P=0.005), and anemia ( OR=8.048, 95% CI: 2.086-31.049; P=0.002) were independent risk factors for SPPH in twin pregnancies during vaginal delivery. Conclusions:Anemia should be actively corrected during twin pregnancy. For twin pregnant women with advanced age, hypertensive disorders in pregnancy, or other risk factors of SPPH, if vaginal delivery is chosen, attention should be paid to the management of labor duration, dynamic assessment of the risk of postpartum hemorrhage, and proactive measures should be taken to ensure a smooth vaginal delivery and effectively reduce the incidence of SPPH.

Objective:To investigate the prognostic factors of ultrasound-guided microwave ablation (MWA) in the treatment of papillary thyroid carcinoma.Methods:The medical records of 97 patients with papillary thyroid carcinoma treated in Shangqiu First People’s Hospital from Jun. 2019 to Dec. 2021 were retrospectively analyzed.All of them were treated with ultrasount-guided MWA,including 38 males and 59 females,aged (48.24±7.86) years. The patients were followed up until Oct. 2024, and the prognosis of the patients was statistically analyzed.According to whether there was new lymph node metastasis or recurrence,the patients were divided into good prognosis group and poor prognosis group.Statistical software SPSS26.0 was used to process the baseline data of the two groups,and the factors affecting the prognosis of papillary thyroid carcinoma were analyzed.Results:Among 97 patients,3 cases were lost to follow-up,73 cases were in good prognosis group and 21 cases were in poor prognosis group.The incidence of multiple lesions,diameter 5-10 mm,close envelope and BRAFV600E mutations in poor prognosis group were 52.38%, 76.19%, 61.90% and 57.14%, respectively, which were higher than 27.40%, 49.32%, 20.55% and 31.51% in good prognosis group ( P<0.05) .TSH level of (2.94±0.61) mlU/L was higher than that of good prognosis group (2.67±0.52) mlU/L ( P<0.05) .Multivariate Logistic regression analysis showed that multiple lesions ( OR=2.915,95%CI:1.073-7.916) ,diameter 5-10mm ( OR=3.289,95% CI:1.090-9.920) , close to the envelope ( OR=6.283,95% CI:2.203-17.917) and BRAFV600E mutations ( OR=2.899,95% CI:1.071-7.843) were independent risk factors for poor prognosis in ultrasound-guided MWA treatment of isthmic papillary carcinoma ( P<0.05) .The ROC curve showed that the AUC value of the four combinations was 0.895,which was significantly higher than the number of lesions (0.625) , tumor size (0.634) , close envelope (0.707) and BRAFV600E mutation (0.628) . Conclusion:Multiple lesions,5-10mm in diameter,close envelope and BRAFV600E mutation are the factors affecting the poor prognosis of patients with isthmic papillary carcinoma treated with ultrasound-guided MWA,and the combination of the four factors is more effective in predicting the prognosis of patients with papillary thyroid carcinoma.

This study aims to simultaneously detect three antibiotic resistance genes(blaNDM,mcr-1 and cfr).A triplex fluorescence quantitative PCR method was established.Plasmids,primers and probes were designed and optimized.The method could specifically detect blaNDM,mcr-1 and cfr,but not other antibiotic resistance genes.The R2 of the standard curves of the three antibiotic re-sistance genes were all greater than 0.999,and the coefficients of variation were all lower than 1％.The lowest detection limits of the plasmids were 1 × 102 copies/μL.This method was used to de-tect 800 bacterial samples.The results showed that 32 samples contained mcr-1 gene,40 samples contained blaNDM gene,2 samples contained cfr gene,8 samples contained both mcr-1 and blaNDM genes.There were no samples carrying three antibiotic resistance genes detected.The results indica-ted that the triplex fluorescence quantitative PCR method established in this experiment had the advantages of high sensitivity,specificity and stability.It was suitable for rapid detection of blaNDM,mcr-1 and cfr antibiotic resistance genes in clinical practice.It provided a convenient and quick method basis for the detection of antibiotic resistance genes.

Objective:To construct a sizeable naive human Fab phage display antibody library to screen high-affinity specific antibodies in vitro. Methods:Total RNA was extracted from peripheral blood mononuclear cells (PBMCs) of 126 healthy individuals, subsequently reverse-transcribed into cDNA, and used as a template. PCR amplification was performed to obtain the V H from IgG, IgM and light chain κ, λ, separately, with the initial PCR products serving as templates for a second round of PCR. Overlap extension PCR was employed to generate fragments of the κ and λ light chains. These fragments were ligated with the phage vector pNC3, which harbors the variable region 1 of the heavy chain, to construct a recombinant phage plasmid. This plasmid was then electroporated into competent Escherichia Coli TG1 cells to establish a naive human Fab phage display antibody library. One hundred clones were randomly selected for identification and sequencing, and antibody gene polymorphisms were analyzed using the IMGT database and MAFFT software. Recombinant α-hemolysin from Staphylococcus aureus was utilized to screen Fab antibody fragments through biopanning of the antibody library, followed by random selection of phage ELISA-identified clones. The positive clones (antigen A450∶blank control A450≥2.1) were sequenced. Results:Two large naive Fab phage display antibody libraries were successfully constructed, in which the capacity of κ and λ chain antibody libraries were 1.25×10 11 and 1.54×10 11, respectively. The titers for two antibody libraries were 6.04×10 13 CFU/ml and 3.50×10 13 CFU/ml. The positive transformation insertion rates for κ and λ chain antibody libraries were 96% (96/100) and 100% (100/100), respectively. Sequence analysis revealed that all antibody sequences were unique. The amino acid sequences in the skeletal region were relatively conserved. In contrast, significant variations in the length of the complementarity determining region (CDR) were found, and the diversity of amino acid sequence of the complementary determining region was high, especially the CDR3. Analysis using the IMGT database indicated that the sequences exhibited a broad distribution across variable-diversity-joining gene families. After six rounds of panning, specific phage antibodies enrichment targeting α-hemolysin were achieved. A total of 142 monoclonal antibodies were sequenced, yielding 8 distinct Fab antibody sequences. Conclusion:This study successfully constructed two naive human Fab phage display antibody libraries with large capacity and good diversity, which can be used for screening human antibodies for serum epidemiology.