Objective To investigate the effects of zalcitabine(ddC),a mitochondrial DNA polymerase γ(POLG)inhibitor,on the migration,invasion,and to preliminarily explore mitochondrial biogenesis of human tri-ple-negative breast cancer MDA-MB-231 cells.Methods The effect of ddC on cell viability was detected using the MTT assay.The migration and invasion abilities of the cells were evaluated using the cell scratch and Transwell in-vasion assays.Cell apoptosis was determined using flow cytometry and a V-FITC/PI cell apoptosis detection kit.The protein expression of POLG,NADH dehydrogenase subunit Ⅰ(NADH1),NADH dehydrogenase subunit Ⅱ(NADH2),ATP synthase subunit 6(ATPase6),cytochrome c oxidase subunit Ⅰ(COX-1)and cytochrome c ox-idase subunit Ⅲ(COX-3)were determined using Western blot.The POLG mRNA level and mtDNA copy number were determined using qPCR.The mitochondrial content and ATP levels were determined using MitoTracker Green fluorescent probe staining and an ATP determination kit.MDA-MB-231 cells were transfected with pcDNA3.1-EG-FP-POLG plasmids to overexpress POLG.The inhibitory effects of ddC on cell migration and invasion were detected in POLG-overexpressed MDA-MB-231 cells.Results POLG expression was higher in MDA-MB-231 cells than in normal mammary epithelial cells(MCF-10A)(P＜0.01).ddC inhibited cell viability in a dose-dependent man-ner.ddC inhibited the migration(P＜0.01)and invasion(P＜0.01)of MDA-MB-231 cells;however,it dis-played no significant inhibitory effects on cell viability in normal mammary epithelial cells(MCF-10A)at the same concentration.ddC downregulated the protein(P＜0.01)and mRNA(P＜0.01)levels of POLG,reduced mtD-NA copy number(P＜0.01)and downregulated mtDNA-coded NADH1,NADH2,ATPase6,COX-1 and COX-3 protein expression(P＜0.01)in MDA-MB-231 cells.Furthermore ddC inhibited mitochondrial content(P＜0.01)and ATP(P＜0.01)levels in MDA-MB-231 cells.POLG overexpression increased the migration(P＜0.05)and invasion(P＜0.05)abilities of MDA-MB-231 cells,while ddC did not significantly inhibit the migra-tion and invasion abilities of MDA-MB-231 cells overexpressing POLG.Conclusion ddC downregulates POLG ex-pression in MDA-MB-231 cells and inhibits mitochondrial biogenesis and ATP levels,thereby inhibiting the migra-tion and invasion of MDA-MB-231 cells.

Objective To investigate the risk factors for portopulmonary hypertension(POPH)in liver cirrhosis,and to construct a noninvasive predictive model.Methods A retrospective analysis was performed for the clinical data of 310 cirrhotic patients with portal hypertension who were hospitalized in The Third Affiliated Hospital of Hebei Medical University from January 2013 to August 2022,and according to whether pulmonary artery systolic pressure was≥40 mmHg on ultrasound,the patients were divided into POPH group with 31 patients and non-POPH group with 279 patients.The independent-samples t test was used for comparison of normally distributed continuous data between two groups,and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups;the chi-square test or the Fisher's exact test was used for comparison of categorical data between two groups.A binary Logistic regression analysis was used to determine the independent risk factors for POPH,and a nomogram prediction model was constructed.The Bootstrap resampling method was used for internal validation,and C-index and calibration curve were used to assess the discriminatory ability and consistency of the model.The rms package was used to plot the nomogram.Results Compared with the non-POPH group,the POPH group had a significantly younger age,a significantly higher proportion of women or patients with hepatic encephalopathy or Child-Pugh class C disease,and significantly higher levels of direct bilirubin,Model for End-Stage Liver Disease score,albumin-bilirubin(ALBI)score,international normalized ratio,prothrombin time,FIB-4 index,LOK score,and Forns index,as well as significantly lower levels of serum albumin,alanine aminotransferase,gamma-glutamyl transpeptidase,hemoglobin,total cholesterol,and triglycerides(all P<0.05).The multivariate analysis showed that sex(odds ratio[OR]=0.172,95%confidence interval[CI]:0.064-0.462,P<0.001),age(OR=0.944,95%CI:0.901-0.989,P=0.016),ALBI score(OR=3.091,95%CI:1.100-8.687,P=0.032),and hepatic encephalopathy(OR=3.466,95%CI:1.331-9.031,P=0.011)were independent risk factors for POPH.A predictive model for POPH in liver cirrhosis was established based on the above independent risk factors,with a C-index of 0.796(95%CI:0.701-0.890),suggesting that the model had good discriminatory ability,and the calibration curve showed that the model had good calibration ability,suggesting that the model had certain predictive efficacy.Conclusion Young female individuals,elevated ALBI score,and comorbidity with hepatic encephalopathy are independent risk factors for POPH in patients with liver cirrhosis,and the predictive model established based on these factors has a certain clinical application value.

Objective To investigate the main dietary patterns of children in urban areas of Maanshan City, and to explore the association between dietary patterns and subthreshold autism trait (SAT), attention-deficit/hyperactivity disorder (ADHD) symptoms and comorbid behaviors in 3-year-old children. Methods Based on the birth cohort of Maanshan Maternal and Infant Health from June 2015 to June 2016，regular physical examinations were conducted from 4 to 2 to 1, and follow-up was conducted until the age of 3. A semi-quantitative food frequency questionnaire was employed to assess dietary intakes. SAT and ADHD symptoms were assessed by Clancy autism behavior scale (CABS) and the 10-item Chinese version of the Conners Abbreviated Symptom Questionnaire (C-ASQ). Social-demographic information was also collected. Results The detection rates of SAT, ADHD symptoms, and comorbidity were 11.03%, 5.28%, and 2.71%, respectively. The older the father, the higher the mother's education level, and the higher the per capita monthly income of the family, the lower the SAT detection rate (P<0.05). The higher the father's educational level, the lower the detection rate of ADHD symptoms (P<0.05). The older the mother and the higher the education level, the lower the detection rate of comorbid behavior in their children (P <0.05). After adjusting for demographic influencing factors, the low intake level was used as a reference. The vegetarian type with moderate intake level was negatively correlated with SAT, while the processed food type with high intake level was positively correlated with SAT. The vegetarian type with high intake level was negatively correlated with ADHD symptoms, while the snack type with high intake level was positively correlated with ADHD symptoms. The vegetarian type with high intake level was negatively correlated with comorbidity. Conclusion Unhealthy dietary patterns are related to children's SAT and ADHD symptoms. Correcting children's unhealthy eating patterns may help reduce children's behavior problems.

Objective:To explore the relationship between the maternal dietary patterns during pregnancy and the early childhood BMI change trajectory.Methods:The subjects were 1 241 pairs of pregnant women and their children in Ma'anshan maternal and infant health cohort. The food frequency questionnaire was used to collect the maternal diet data during pregnancy. The cohort children were followed up at birth, month 3, 6, 12, 18 and 24, respectively. The body height and weight data of the cohort children were collected. The principal component analysis was used to determine the categories of maternal dietary patterns during pregnancy, group-based multi-trajectory modeling was used to fit the early childhood BMI change trajectory, and the multiple classification logistic regression model was used to evaluate the relationship between the maternal dietary patterns during pregnancy and the early childhood BMI change trajectory.Results:The maternal dietary patterns during pregnancy included protein type, healthy type, vegetarian type, processing type and beverage type, which could explain 50.04% of the total dietary variation. Among them, the protein type, main dietary pattern, could explain 21.34% of the total dietary variation. The early childhood BMI change trajectory was from thinnish stature to average stature, then to mild obesity, accounting for 42.9%, 45.6% and 11.5% respectively. After controlling the potential confounding factors, it was found that there was a statistical correlation between healthy type and beverage type of maternal dietary patterns during pregnancy and early childhood BMI change trajectory ( P<0.05). Comparison of change trajectories between thinnish type and average stature type, children in the low-level group of healthy diet pattern tended to have a thinnish type change trajectory in early life ( OR=1.286, 95% CI: 1.002-1.651). Comparison of change trajectories between mild obesity type and average stature type, children in the high-level group of beverage diet pattern tended to have a mild obesity type change trajectory in early life ( OR=0.565, 95% CI: 0.342-0.935). The other dietary patterns had no statistical correlation with the early childhood BMI change trajectory. Conclusions:Maternal dietary patterns during pregnancy can affect the early childhood BMI change trajectory, and the low-level healthy type diet is an independent risk factor for thinnish type change trajectory, and the high-level beverage type diet is an independent risk factor for the mild obesity type change trajectory.

Cadmium exposure during pregnancy is a non-negligible public health problem which may increase the risk of shortened telomere length in newborns and cardiovascular metabolic health damage in children, and has attracted attention from many researchers in recent years. This article reviewed recent studies both domestically and internationally on the associations among cadmium exposure during pregnancy, shortened telomere length in newborns, and cardiovascular metabolic abnormalities in children, and briefly outlined possible mechanisms of shortened telomere length in newborns by cadmium exposure during pregnancy. Current research results showed that cadmium exposure during pregnancy is related to shortened telomere length in newborns and cardiovascular metabolic abnormalities in children, and shortened telomere length in newborns is also related to cardiovascular metabolic abnormalities in children. It suggested that telomere length in newborns may be a biomarker reflecting cardiovascular metabolic abnormalities in children caused by cadmium exposure during pregnancy. In addition, the current potential mechanisms of cadmium exposure during pregnancy accelerating neonatal telomere length shortening include inflammatory reaction, mitochondrial dysfunction, antioxidant consumption/antioxidant enzyme inactivation, and DNA methylation, and these biological mechanisms are associated with cardiovascular metabolic abnormalities through certain factors, such as obesity, elevated blood pressure, impaired fasting blood glucose, and dyslipidemia in children, suggesting that cardiovascular metabolic abnormalities in children may be programmed in early life, but there are still few relevant studies. In the future, research should be conducted on the association among cadmium exposure during pregnancy, telomere length, and offspring cardiovascular metabolism, as well as possible mediating efficacy and related biological mechanisms of telomere length, aiming to provide early-life biological information for the prevention of cardiovascular and metabolic diseases.

Objective: To explore the effect of hypoxia inducible factor 1α(HIF-1α)on tamoxifen resistance in breast cancer MCF-7,and to established a human breast cancer cell line(MCF-7/TR)with 4-hydroxy tamoxifen(4-OHT)resistance. Methods: 4-OHT was an active form of tamoxifenin vivo, resistant breast cancer cells MCF-7/TR were establishedin vitrousing 4-OHT. MTT assay was used to detect the effect of 4-OHT on the MCF-7 and MCF-7/TR cells, and the drug resistance multiple was detected. Western blot was used to detect the expression level of HIF-1α protein in MCF-7 and MCF-7/TR cells. The effects of HIF-1α inhibitor(LW6) or siRNA HIF-1α on MCF-7/TR cells and 4-OHT on MCF-7 cells treated with HIF-1α stabilizer(FG-4592) were detected by MTT and flow cytometry AV/PI staining. Results: The results showed that the MCF-7/TR was successfully constructed, and the drug resistance ratio was(5.56±0.80). Compared with MCF-7 cells, MCF-7/TR cells had higher expression of HIF-1α protein and it was up-regulated after tamoxifen treatment. After giving LW6 or silencing the expression of HIF-1α,the down-regulation of HIF-1α expression enhanced the inhibitory effect of 4-OHT on MCF-7/TR cells. After treatment of FG-4592, the expression level of HIF-1α in breast cancer cells MCF-7 was up-regulated, and hindered the inhibition effect of tamoxifen on MCF-7 cells. Conclusion The above results indicate that HIF-1α plays an important role in 4-OHT resistant breast cancer, and targeting HIF-1α may be an effective way to increase the sensitivity of MCF7/TR cells to tamoxifen.

Objective : To investigate the association between adiposity rebound and metabolic abnormalities in pre- schools． Methods : A prospective cohort study was designed on the basis of the Maanshan birth cohort． Venous blood samples were collected at 5 to 6 years of age to detect metabolic indicators．2022 children aged 0 to 6 years with ≥8 consecutive measurements were enrolled. χ2 test and Logistic regression model were used to analyze the data. Results : The detection rate of abnormal metabolism in preschool children was 16. 9% ，and the risk of meta- bolic abnormalities in preschool children with high BMI level at the AR time point and earlier AR time phase was 2. 59 and 1. 82 times that of the normal group respectively． Conclusion High AR level and earlier AR phase can increase the prevalence of metabolic abnormalities in preschool children.

Objective:To investigate the effects of maternal subclinical hypothyroidism (SCH) on preschoolers′ behavioral problems.Methods:Based on the Ma′ anshan Birth Cohort, pregnant women who had their first antenatal checkup in Maternal and Child Health Center in Ma′ anshan were recruited from May 2013 to September 2014. Data on demographic, obstetric information, and maternal exposure were collected. Women′s fasting venous blood in the first, second, and third trimesters of pregnancy was collected. The levels of thyroid hormones [thyrotropin (TSH), free thyroxine (FT 4)] and thyroid autoantibodies [thyroid peroxidase antibody (TPOAb), thyroglobulin antibody (TgAb)] in maternal blood were retrospectively detected by electrochemiluminescence immunoassay. Preschoolers′ behavioral problems were assessed by Achenbach Child Behavior Checklist (CBCL/1.5-5). Poisson regression models were adopted to examine the effect of maternal SCH on preschoolers′ internalizing and externalizing problems and the critical period. Results:In this study, the reference of maternal thyroid indexes was established (between 2.5 th and 97.5 th percentile). The reference of TSH in the first, second, and third trimester of pregnancy was 0.04-4.90 μIU/mL, 0.75-6.08 μIU/mL, and 0.58-5.59 μIU/mL respectively; and the reference of FT 4 was 13.19-23.27 pmol/L, 9.14-15.32 pmol/L, and 9.53-17.45 pmol/L respectively. In the first, second, and third trimester of pregnancy, the prevalence of SCH was found to be 2.0% (25/1 224), 1.6% (19/1 218), and 1.7% (21/1 220), respectively. After adjusting for confounding factors, maternal SCH in the first trimester was associated with the risk of anxiety and depression in preschool children ( OR=3.06, 95% CI 1.05-8.98). Maternal SCH in the second trimester was found to be associated with the risk of overreaction in preschool children ( OR=2.65, 95% CI 1.13-6.21). Conclusions:The establishment of thyroid hormones reference range for pregnant women in Ma′ anshan area is beneficial to the screening, diagnosis, and treatment of thyroid diseases during pregnancy in this area. Maternal SCH during pregnancy is associated with increased risk of behavioral problems in preschool children. In the first trimester, maternal SCH was associated with preschoolers′ anxiety and depression, and in the second trimester, maternal SCH was associated with preschoolers′ emotional reactivity.

Objective:To explore the protective effects of breastfeeding on behavioral problems at 4 years in children born to mothers with gestational diabetes mellitus (GDM).Methods:Based on the Ma' anshan Birth Cohort (MABC) study, 305 GDM women and their children were recruited in this study from Ma' anshan Maternal and Child Health Care Hospital from May 2013 to September 2014. Total breastfeeding duration was followed up at 42 d, 3, 6, 12, and 18 months postpartum as well as the breastfeeding intensity within 6 months. All the subjects were divided into breastfeeding group ( n=256, including exclusive breastfeeding and mixed feeding) or bottle feeding group ( n=49). Internalizing and externalizing behavioral problems at age 4 were assessed using Achenbach Child Behavior Checklist (CBCL/1.5~5) and their association with breastfeeding were analyzed using robust Poisson regression. Controlling false discovery rate was applied for multiple test correction. Results:Compared with bottle feeding, breastfeeding was a protective factor for depression in children ( RR=0.23, 95% CI: 0.05-0.98, q=0.048) when the duration was 4-5 months; for somatic complaints ( RR=0.36, 95% CI: 0.14-0.95, q=0.047) and anxiety ( RR=0.19, 95% CI: 0.06-0.62, q=0.010) with a breastfeeding duration of 6-11 months; and for depression ( RR=0.46, 95% CI: 0.25-0.83, q=0.039) and anxiety ( RR=0.12, 95% CI: 0.03-0.49, q=0.006) with a breastfeeding duration of 12 months and above. Compared with bottle feeding within 6 months, mixed feeding had a protective effect on somatic complaints ( RR=0.29, 95% CI: 0.13-0.64, q=0.026) and anxiety ( RR=0.18, 95% CI:0.07-0.52, q=0.002). Conclusions:The findings suggested that breastfeeding had a protective effect on behavioral problems at age 4 in children exposed to GDM. Women with GDM should be encouraged to breastfeed.

Objective:To investigate the influence and critical period of pregnancy-related anxiety during pregnancy on the neurobehavioral development of infants.Methods:The subjects of this study were derived from the Ma′anshan Birth Corhot. From May 2013 to September 2014, a total of 3 474 pregnant women who registered in Ma ′anshan Maternal and Child Health Care Center were enrolled in the study. A total of 2 242 mother-infant pairs who completed three times assessments of maternal anxiety and at least once assessment of infants′ neurobehavioral development were included in the final analysis. Maternal pregnancy-related anxiety was assessed by the Pregnancy-Related Anxiety Questionnaire during the first, second and third trimesters of pregnancy. When their children were at 6 and 18 months, their neurobehavioral development was evaluated using the Ages & Stages Questionnaire-China. The influence of maternal pregnancy-related anxiety on the neurobehavioral development of infants was analyzed by bi-nominal logistic regression.Results:The age of 2 242 pregnant women was (26.62±3.65) years, and the proportion of boys, low birth weight and exclusive breastfeeding for 6 months was 50% (1 120/2 242), 1.7% (38/2 242) and 11.5% (252/2 191), respectively. The detection rates of pregnancy-related anxiety during the first, second and third trimester were 24.9% (558), 28.6% (642) and 30.3% (674), respectively. After controlling confounding variables and other two trimester′s anxiety, only pregnancy-related anxiety during the third trimester (not first or second trimester) significantly increased the risk of developmental delay in the domain of communication (relative risk, RR = 3.52, 95% confidence interval, CI: 1.89-6.58) and personal-social ( RR=2.46, 95% CI: 1.10-5.49) at the 6 months of age, as well as in the domain of fine motor ( RR=2.07, 95% CI: 1.11-3.85), problem-solving domains ( RR=2.31, 95% CI: 1.24-4.31). Conclusion:Maternal pregnancy-related anxiety was associated with the risk of neurobehavioral development of infants, and the third trimester may be the critical period.