Background::Heterotaxy (HTX) is a thoracoabdominal organ anomaly syndrome and commonly accompanied by congenital heart disease (CHD). The aim of this study was to analyze rare copy number variations (CNVs) in a HTX/CHD cohort and to examine the potential mechanisms contributing to HTX/CHD.Methods::Chromosome microarray analysis was used to identify rare CNVs in a cohort of 120 unrelated HTX/CHD patients, and available samples from parents were used to confirm the inheritance pattern. Potential candidate genes in CNVs region were prioritized via the DECIPHER database, and PNPLA4 was identified as the leading candidate gene. To validate, we generated PNPLA4-overexpressing human induced pluripotent stem cell lines as well as pnpla4-overexpressing zebrafish model, followed by a series of transcriptomic, biochemical and cellular analyses. Results::Seventeen rare CNVs were identified in 15 of the 120 HTX/CHD patients (12.5%). Xp22.31 duplication was one of the inherited CNVs identified in this HTX/CHD cohort, and PNPLA4 in the Xp22.31 was a candidate gene associated with HTX/CHD. PNPLA4 is expressed in the lateral plate mesoderm, which is known to be critical for left/right embryonic patterning as well as cardiomyocyte differentiation, and in the neural crest cell lineage. Through a series of in vivo and in vitro analyses at the molecular and cellular levels, we revealed that the biological function of PNPLA4 is importantly involved in the primary cilia formation and function via its regulation of energy metabolism and mitochondria-mediated ATP production. Conclusions::Our findings demonstrated a significant association between CNVs and HTX/CHD. Our data strongly suggested that an increased genetic dose of PNPLA4 due to Xp22.31 duplication is a disease-causing risk factor for HTX/CHD.

Aim To investigate the impact of Cinobu-facini on the proliferation,invasion,and apoptosis of HepG2 cells and the underlying mechanism.Methods The proliferation of HepG2 cells was assessed using the CCK-8 method following treatment with Cinobufaci-ni.The invasion capability of HepG2 cells was evalua-ted through Transwell assay after exposure to Cinobufa-cini.The apoptosis rates of HepG2 cells post Cinobufa-cini intervention were measured using flow cytometry,and the expression levels of VEGF in the culture medi-um of HepG2 cells were determined using enzyme-linked immunoassay.Furthermore,qRT-PCR and Western blot analyses were conducted to assess the im-pact of Cinobufacini on mRNA and protein expression levels related to the CXCL5/FOXD1/VEGF pathway.The interaction between CXCL5 and FOXD1 was inves-tigated via co-immunoprecipitation.Results Cinobufa-cini treatment led to a gradual decrease in HepG2 cell viability in a dose-dependent manner compared to the control group(P＜0.05).Moreover,Cinobufacini sig-nificantly suppressed HepG2 cell invasion(P＜0.05)while enhancing cell apoptosis(P＜0.05).Notably,Cinobufacini exhibited inhibitory effects on the CX-CL5/FOXD1/VEGF pathway,as evidenced by re-duced expression of related mRNA and proteins(P＜0.05).FOXD1 was identified as the binding site of CXCL5.Overexpression of CXCL5 resulted in in-creased proliferation and VEGF secretion by HepG2 cells(P＜0.05),and increased expression of FOXD1 and VEGF(P＜0.05).However,Cinobufacini inter-vention effectively inhibited liver cancer cell prolifera-tion and invasion(P＜0.05),promoted apoptosis(P＜0.05),reduced VEGF secretion by HepG2 cells(P＜0.05),and downregulated the expression of CXCL5 and FOXD1 in HepG2 cells(P＜0.05);but com-pared with the unexpressed group of Cinobufacini,its ability to inhibit cell activity was weakened(P＜0.05),and its ability to inhibit the expression of CX-CL5,FOXD1,and VEGF was weakened(P＜0.05).Conclusion Cinobufacini may inhibit HepG2 cell pro-liferation and invasion and promote HepG2 cell apopto-sis by regulating the CXCL5/FOXD1/VEGF pathway.

Objective:To evaluate the optimization effect of transcutaneous electrical acupoint stimulation (TEAS) combined with moderate sedation with propofol (TEAS-propofol balanced anesthesia) for gastrointestinal endoscopy.Methods:This was a single-blind randomized controlled trial. American Society of Anesthesiologists Physical Status classification Ⅰ-Ⅲ patients, aged 18-64 yr, undergoing elective gastrointestinal endoscopy at the Endoscopic Diagnosis and Treatment Center of the Affiliated Hospital of Qingdao University from May to August 2022, were divided into 2 groups using the block random allocation method: conventional anesthesia group (group C) and TEAS-propofol balanced anesthesia group (group TPB). Patients received moderate sedation with propofol plus routine anesthesia with fentanyl 50 μg in group C. In TPB group, TEAS was performed at bilateral Neiguan, Hegu and Zusanli acupoints before surgery until the end of surgery, and patients received propofol for moderate sedation (Modified Observer′s Assessment of Alertness/Sedation scale score was 3). The efficacy and safety of anesthesia and parameters related to outcomes were observed and recorded.Results:In this study, 66 patients were recruited, with 33 in each group, the failure rate of anesthesia in both groups was 3%, and no reflux or aspiration was found. Compared with group C, no significant changes were found in the patients′ satisfaction on the same day, intraoperative pain response score, incidence of intraoperative adverse reactions (tachycardia, hypertension, bucking and body movement), awake time, consumption of propofol, rate of intraoperative awareness and rate of patients hoping to receive the same anesthesia method again postoperatively ( P>0.05), the patients′ satisfaction was significantly increased on the next day ( P<0.05), the incidence of intraoperative respiratory depression, physician satisfaction, and degree of postoperative dizziness and nausea were significantly reduced ( P<0.05), and the discharge time and time to the complete recovery of normal behavior function was significantly shortened in group TPB ( P<0.05). Conclusions:The combination of TEAS at bilateral Neiguan, Hegu and Zusanli acupoints with moderate sedation using propofol for gastrointestinal endoscopy is not only safe and effective, but also beneficial to the postoperative outcome of patients, and the effect is better than that of conventional anesthesia with propofol and fentanyl.

AIM To explore the effects of praeruptorin A from Suhuang antitussive capsules on cough variant asthma(CVA).METHODS The rats were randomly divided into the normal group,the model group,the dexamethasone group(0.5 mg/kg),the Suhuang antitussive capsules group(7 g/kg)and the low,medium and high dose praeruptorin A groups(15,30 and 60 mg/kg).The rat model of CVA was established by intraperitoneal injection of sensitizer(1 mg/mL ovalbumin and 10 mg/mL aluminum hydroxide)and aerosol inhalation of 1%ovalbumin followed by the corresponding dosing of drugs by gavage initiated on the 14th day.Another 14 days later,the rats had their pathological pulmonary changes observed by HE,Masson and PAS stainings;their number of inflammatory cells in bronchoalveolar lavage fluid(BALF)detected by hematology analyzer;and their levels of IL-4,IL-5,IL-13 and MUC5AC in BALF detected by ELISA.The RAW264.7 cell inflammatory model induced by lipopolysaccharide(LPS)was treated with 4,8,16 μmol/L praeruptorin A or 0.25 mg/mL Suhuang antitussive capsules,respectively.And the cells had their NO level detected by Griess method,and their ROS expression observed using fluorescence microscopy.The detections of the pulmonary and cellular mRNA expressions of IL-6,IL-1β,COX-2,iNOS and PPAR-γ by RT-qPCR;and the protein expressions of p-P65,P65,p-IκBα,IκBα,NLRP3,caspase-1(p20)and IL-1β by Western blot were conducted in both the cells and the rats.RESULTS The in vivo result showed that praeruptorin A reduced the cough frequency(P＜0.01);prolonged the cough latency(P＜0.05,P＜0.01);reduced the number of eosinophils and neutrophils in BALF(P＜0.05,P＜0.01);decreased the levels of IL-4,IL-5,IL-13 and MUC5AC in BALF and the pulmonary mRNA expressions of IL-6,IL-1β,COX-2 and iNOS(P＜0.05,P＜0.01);and decreased the phosphorylation of P65 and IκBα protein and NLRP3,caspase-1(p20)and IL-1β protein expressions(P＜0.05,P＜0.01)as well.The in vitro result showed that praeruptorin A inhibited the release of LPS-induced NO and reduce the ROS level(P＜0.01);decreased the mRNA expressions of IL-1β,COX-2 and iNOS(P＜0.05,P＜0.01);increased PPAR-γ mRNA expression(P＜0.05),and decreased the phosphorylation of P65 and IκBα protein and the expression of NLRP3 protein(P＜0.05,P＜0.01).CONCLUSION Praeruptorin A,one of the main antitussive components of Suhuang antitussive capsules,may improve CVA because of its anti-inflammatory and antitussive role by inhibiting the activation of NF-κB signaling pathway and reducing the expression of NLRP3 inflammatory corpuscles.

Objective Using healthy female reproductive-age rhesus macaques as the research subjects,we explored the correlation between menstrual cycle abnormalities and gut microbiota composition by using 16S rRNA metagenomic sequencing.Methods Twenty-seven healthy female rhesus macaques were divided into regular menstrual and irregular menstrual groups.Fecal samples were collected at follicular phase(FP),ovulation phase(OP)and luteal phase(LP)of the two groups.The structure and diversity of bacterial flora in different physiological periods were analyzed and compared between the two groups.Results At the phylum level,Firmicutes,Bacteroidetes,and Proteobacteria dominated the sample flora in the follicular,luteal,and ovulatory phases of the rhesus macaques in both the regular and irregular groups,with a combined percentage of more than 98% .At the genus level,the genus Prevotella_9,Ruminococcaceae_UCG-002,Lactobacillus,Prevotella_2,Phascolarctobacterium,Ruminococcaceae_UCG-005,Streptococcus,Blautia,Prevotellaceae_NK3B31_group,Rikenellaceae_RC9_gut_group were dominant.In the luteal phase the percentage of Firmicutes was higher in the regular group than in the irregular group,while the opposite was true for Bacteroidetes.Spirochaetes were higher in the regular group than in the irregular group at all 3 stages(P＜0.05).Conclusion There were some differences in intestinal microbial composition between the two groups of macaques with regular and irregular menstrual cycles,which provided some reference for the study of intestinal bacteria and ovulation disorders.

Objective:To investigate the differences in multi-b-value apparent diffusion coefficient (ADC) and exponential apparent diffusion coefficient (eADC) between hepatocellular carcinoma (HCC) and paracancerous liver tissue, distant cancerous liver tissue, and background liver tissues by ultra-high field 3.0T diffusion-weighted (DWI) MRI imaging.Methods:Sixty-eight consecutive HCC cases confirmed by surgical pathology from January 2018 to October 2021 were enrolled and divided into a cirrhosis ( n=39) and a non-cirrhosis group ( n=29) according to the presence or absence of cirrhosis.The average ADC and eADC of liver tissues of paracancerous (including proximal and distal), distant cancerous, and background were measured by DWI images with diffusion sensitivity factors (b) of 50, 100, 400, 600 s/mm 2, and 1 000 s/mm 2, respectively. The Kruskal-Wallis H test and Bonferroni method were used to test the differences between the measured values of the five tissues. The statistical differences were used to evaluate the diagnostic efficacy of the five tissues by parametric receiver operating characteristic (ROC) curve and area under the curve (AUC). Results:The comparison of average ADC and eADC among five types of tissues in the liver cirrhosis group showed that the average ADC and eADC measured at b values of 50, 100, 400, and 600 s/mm 2 had statistically significant differences (adjusted P<0.005) between cancerous and proximal paracancerous, distal paracancerous, distant cancerous, and background liver tissue, as well as the average ADC measured at b=1 000 s/mm 2 between cancerous and proximal paracancerous tissue. The average ADC and eADC in the non-cirrhosis group had statistically significant differences (adjusted P<0.005) between cancerous and proximal paracancerous, distant paracancerous, distant cancerous, and background liver tissue measured at b values of 50, 100, and 400 s/mm 2, respectively. The average ADC and eADC measured at b=600 s/mm 2 showed statistically significant differences (adjusted P<0.005) between cancerous and proximal paracancerous, distal paracancerous, and distant cancerous liver tissue, as well as the average ADC measured at b=1 000 s/mm 2 between cancerous and distal paracancerous, and distant cancerous liver tissue. The average ADC and eADC in the cirrhosis group had no statistically significant difference between the proximal paracancerous and the distant cancerous, as well as the background liver tissue measured at b-values of 50, 100, 400, 600, and 1 000 s/mm 2, respectively (adjusted P>0.005), while there were statistically significant differences (adjusted P<0.005) in the average ADC values in the non-cirrhosis group between the proximal paracancerous and the distant paracancerous and background liver tissues at b=50 s/mm 2, as well as the average ADC and eADC values between the proximal paracancerous and the distant liver tissues at b=100 s/mm 2. The average ADC and eADC values measured in the cirrhosis group and non-cirrhosis group had no statistically significant difference between the distant paracancerous, distant cancerous, and background liver tissue (adjusted P>0.005). The efficacy of average ADC and eADC in distinguishing five types of tissues (cancerous and proximal paracancerous, distant paracancerous, distant cancerous, and background liver tissue) showed that in the cirrhosis group, the diagnostic efficacy was best at b=50 s/mm 2. The area under the ROC curve (AUC) of average ADC was 0.815, 0.828, 0.855, and 0.855, respectively, and the AUC of average eADC was 0.815, 0.830, 0.856, and 0.855, respectively. The diagnostic efficacy was best in the non cirrhosis group at b=100 s/mm 2, with average ADC AUCs of 0.787, 0.823, 0.841, and 0.821, and average eADC AUCs of 0.836, 0.874, 0.893, and 0.873, respectively. The AUC of the average ADC in the non-cirrhosis group for distinguishing between proximal paracancerous and distant cancerous liver tissues, as well as proximal paracancerous and background liver tissues, with b=50 s/mm 2, were 0.605 and 0.604, respectively. The average AUC of ADC and eADC for distinguishing between proximal paracancerous and distant liver tissues with b=100 s/mm 2 were 0.619 and 0.620, respectively. Conclusion:The average ADC and eADC measured by multiple b-values are helpful in distinguishing HCC from proximal paracancerous, distal paracancerous, distant-cancerous, and background liver tissues in patients with cirrhosis and non-cirrhosis, while the average ADC and eADC at b=50 s/mm 2 and 100 s/mm 2 exhibit differences between the proximal paracancerous from the distant cancerous liver tissue and background liver tissue in patients with non-cirrhosis.

Objective To construct time-series by adopting autoregressive integrated moving average(ARIMA)model for analyzing the trend and genotype characteristics of single-center human papillomavirus(HPV)infection in Tianjin area.Methods A total of 7 236 female patients who underwent HPV testing in a hospital from January 2018 to December 2022 were selected.HPV infection status and genotype distribution in Tianjin area from 2018 to 2022 were compared.ARIMA model time-series was constructed,and model fitting was analyzed.The number of HPV infections in 2023 was predicted and compared with the actual occurrence,the predictive performance of the model was evaluated.Results HPV infection rate in Tianjin area from 2018 to 2022 was 14.41％,with the highest rate(15.47％)in the age group of 31-40 years.Among the positive specimens,the proportion of single type HPV infection was the highest,accounting for 73.54％(767/1 043),with high-risk HPV being the main type.The highest infection rates of low-risk and high-risk types were type HPV-6(2.59％)and type HPV-16(16.06％),re-spectively.ARIMA model was constructed,and the optimal model was ARIMA(0,1,2)(0,1,1)12,with akaike in-formation criterion(AIC)and bayesian information criterion(BIC)values of 3.877 and 4.005,respectively.There was no statistical significance in Ljung-Box Q=8.828 showed by white noise test(P＞0.05).The number of HPV infection in 2023 was predicted by the model.The overall trend of the actual value and the predicted value was basi-cally consistent,RMSE,MAPE and MAE of the model were 6.289,34.149 and 4.706,respectively,suggesting that the model had a good prediction effect.Conclusion Among the female population in Tianjin area,HPV infec-tion is mainly caused by single,high-risk type,with HPV-16 having the highest infection rate.There is seasonal variation in HPV infection in Tianjin.ARIMA model has good prediction effect on the prevalence trend of HPV in-fection,which is suitable for short-term prediction.

Objective:To investigate the feasibility of a three-dimensional no new U-Net (3D nnU-Net) deep learning (DL) network for the automatic segmentation of colorectal cancer (CRC) based on abdominal CT images.Methods:This was a cross-sectional study. From January 2018 to May 2023, a total of 2180 primary CRC patients, confirmed by pathology at the Guangdong Provincial Hospital of Traditional Chinese Medicine (center 1, n=777), Nanfang Hospital, Southern Medical University (center 2, n=732), and Sun Yat-sen Memorial Hospital (center 3, n=671), were enrolled in this retrospective study. The baseline abdominal CT examination of each patient was conducted using CT equipment from 7 different models across 4 vendors, at the 3 centers, encompassing both the arterial phase (AP) and venous phase (VP). Two radiologists manually delineated the volume of interest to circumscribe the entire tumors in dual-enhanced phase CT images. The CT data of CRC patients from center 1 and center 3 were merged and divided into a training set ( n=1 159) and a validation set ( n=289) using a weighted random method with a ratio of 4∶1. The patients from center 2 were used as an independent external test set ( n=732). The 3D nnU-Net segmentation model was trained and tested. Using manually annotated label data as the benchmark, segmentation performance of the model was evaluated based on different phases and tumor locations. The segmentation coverage rate (SCR), Dice similarity coefficient (DSC), recall (REC), precision (PRE), F1-score, and 95% Hausdorff distance (HD 95) were calculated. The mean manual segmentation time and the mean automatic time were compared using independent samples t-test. Results:In the independent external test set, the performance of the 3D nnU-Net model based on the AP CT images was superior to that based on the VP CT images. On the AP images, the SCR, DSC, REC, PRE, F1-score, and HD 95 were 0.865, 0.714, 0.716, 0.736, 0.714, and 27.228, respectively; on the VP images, they were 0.834, 0.679, 0.710, 0.675, 0.679, and 29.358, respectively. The model achieved the best performance on right-sided colon cancer, with SCR, DSC, REC, PRE, F1-score, and HD95 on the AP CT images at 0.901, 0.775, 0.780, 0.787, 0.775, and 21.793, respectively. Next were left-sided colon cancer and rectal cancer, while the segmentation performance for transverse colon cancer was the worst (SCR, DSC, REC, PRE, F1-score, and HD 95 were 0.731, 0.631, 0.641, 0.630, 0.631 and 38.721, respectively). The automatic segmentation time on a single phase was (1.0±0.3) min, while the manual segmentation time was (17.5±6.0) min ( t=128.24, P<0.001). Conclusions:After training and validating on a dataset from multiple centers with various CT scanner vendors, the 3D nnU-Net DL model demonstrates the capability to automatically segment CRC based on abdominal CT images, while also showcasing commendable robustness and generalization ability.

Objective:To investigate the clinical features and prognosis of male with anti-melanoma differentiation-associated gene 5 (MDA5) autoantibody.Methods:The clinical data of 246 patients with DM and anti-MDA5 autoantibodies hospitalized by Jiangsu Myositis Cooperation Group from 2017 to 2020 were collected and retrospectively analyzed. Chi-square test was performed to compared between counting data groups; Quantitative data were expressed by M ( Q1, Q3), and rank sum test was used for comparison between groups; Single factor survival analysis was performed by Kaplan-Meier method and Log rank test; Cox regression analysis were used for multivariate survival analysis. Results:①The male group had a higher proportion of rash at the sun exposure area [67.1%(47/70) vs 52.8%(93/176), χ2=4.18, P=0.041] and V-sign [50.0%(35/70) vs 30.7%(54/176), χ2=8.09, P=0.004] than the female group. The male group had higher levels of creatine kinase [112(18, 981)U/L vs 57 (13.6, 1 433)U/L, Z=-3.50, P<0.001] and ferritin [1 500 (166, 32 716)ng/ml vs 569 (18, 14 839)ng/ml, Z=-5.85, P<0.001] than the female group. The proportion of ILD [40.0%(28/70) vs 59.7%(105/176), χ2=7.82, P=0.020] patients and the red blood cell sedimentation rate[31.0(4.0, 101.5)mm/1 h vs 43.4(5.0, 126.5)mm/1 h, Z=-2.22, P=0.026] in the male group was lower than that of the female group, but the proportion of rapidly progressive interstitial lung disease (PR-ILD) [47.1%(33/70) vs 31.3%(55/176), χ2=5.51, P=0.019] was higher than that of the female group. ②In male patients with positive anti-MDA5 antibodies,the death group had a shorter course of disease[1.0(1.0, 3.0) month vs 2.5(0.5,84) month, Z=-3.07, P=0.002], the incidence of arthritis [16.7%(4/24) vs 42.2%(19/45), χ2=4.60, P=0.032] were low than those in survival group,while aspartate aminotransferase (AST)[64(22.1, 565)U/L vs 51(14,601)U/L, Z=-2.42, P=0.016], lactate dehydrogenase (LDH) [485(24,1 464)U/L vs 352(170, 1 213)U/L, Z=-3.38, P=0.001], C-reactive protein (CRP) [11.6(2.9, 61.7) mg/L vs 4.95(0.6, 86.4) mg/L, Z=-1.96, P=0.050], and ferritin levels [2 000(681, 7 676) vs 1 125 (166, 32 716)ng/ml, Z=-3.18, P=0.001] were higher than those in the survival group, and RP-ILD [95.8%(23/24) vs 22.2%(10/45), χ2=33.99, P<0.001] occurred at a significantly higher rate. ③Cox regression analysis indicated that the course of disease LDH level, and RP-ILD were related factors for the prognosis of male anti-MDA5 antibodies [ HR (95% CI)=0.203(0.077, 0.534), P=0.001; HR (95% CI)=1.002(1.001, 1.004), P=0.003; HR (95% CI)=95.674 (10.872, 841.904), P<0.001]. Conclusion:The clinical manifestations of male anti-MDA5 antibody-positive patients are different from those of female. The incidence of ILD is low, but the proportion of PR-ILD is high. The course of disease, serum LDH level, and RP-ILD are prognostic factors of male anti-MDA5 antibody-positive patients.

Objective:To identify micronuclei through the cytochalasin B blocking micronucleus method-based assay using scanning microscope, combined with the slide scanning software Metafer 4 and, accordingly, to establish a dose-response relationship between the dose of 60Co γ-rays and the frequency of micronuclei in human peripheral blood lymphocytes using artificial intelligence-based color recognition. Methods:Blood samples were collected from four healthy individuals (two men and two women) and were then exposed to varying doses of 60Co γ-ray radiation (0, 0.25, 0.5, 1, 2, 3, 4, 5 Gy) at a dosage rate of 0.74 Gy/min. Micronucleus slides were prepared as per the GBZ 128-2023 standard. The numbers of binuclear cells and micronuclei were recorded using an artificial intelligence-based color recognition analysis system. The dose-response curve was determined through fitting using the CABAS software. Then, the doses to both independent samples were estimated based on the curve. Results:Within a dose range of 0 to 5 Gy, the fitted micronucleus dose-response curve aligned with a quadratic polynomial model, with a regression equation of y = 0.032 1 D2+ 0.023 7 D+ 0.012 7 ( D denoting the dose, correlation coefficient R2=0.998). The dose estimations from the validation samples closely corresponded to the actual irradiation doses. Conclusions:Establishing the micronucleus dose-response curve provides a feasible method and basis for the rapid and accurate estimation of radiation biological doses in laboratory automation.