Objective To prepare VP1 protein vaccine of Coxsackievirus A16(CA16) and evalu-ate immunngenicity the subunit vaccines of Coxsackievirus (VP1), and to establish foundation for studying CA16 vaccine. Methods CA16 VP1 was amplified by RT-PCR and cloned into pFastBac HT A plasmid, recombinated with Bacmid DNA by transposition reaction and then transfected Sf9 cell, mixed with adjuvant AI(OH)_3. After immunization BALB/c mice, evaluating immune effectiveness after booster injections 2 weeks. Results The expressed protein was analyzed by SDS-PAGE and Western blot, mice immunized with CA16 (VP1) both induced specific IgG antibody and neutralization antibody. The best immunization antigen was 20 μg, IgG antibody was 1: 1600, neutralization antibody was 1:250, typical Th1/Th2 immune response was determined by lymphocyte proliferation assay and cytokine analysis. Conclusion The CA16 VP1 gene was cloned successfully and expressed in Sf9 insect cells, CA16 VP1 protein vaccine induced both humoral and cellular immune response, to lay solid foundation for further study on CA16 vaccine.

Objective: To investigate the effect of caffeine citrate administering at different time on outcome and neurodevelopment of premature infants. Methods: A total of 113 preterm infants with gestational age less than 32 weeks and birth weight less than 1 500 g who were hospitalized and treated in the neonatal intensive care unit from January 2018 to June 2018 were enrolled.According to the time when caffeine citrate treatment was started, they were divided into early treatment group(≤1 days) with 53 infants and late treatment group(1 to 10 days) with 60 infants.A retrospective analysis was performed for their clinical data.The perinatal conditions, treatment process and clinical outcomes of the two groups were compared and the neurological development was followed up at 12 months old. Results: Compared with the late treatment group, the early treatment group had a significantly shorter durations of mechanical ventilation time, oxygen therapy time, hospitalization days and a significantly lower incidence of bronchopulmonary dysplasia, patent ductus arteriosusand intraventricular hemorrhage or periventricular leukomalacia, and there were significant differences between two groups(P<0.05, respectively). The neonatal behavioral neurological assessment score in the early treatment group was higher than that in the late treatment group at 40 weeks of gestational age, and there was significant difference between two groups(P<0.05). The mental developmental index at 3 months of corrected age, the mental developmental index and psychomotor developmental index at 12 months were significantly better in the early treatment group than those in the late treatment group, and there were significant differences between two groups(P<0.05, respectively). Conclusion Early use of caffeine citrate can improve the outcome of premature infants and improve the prognosis of nervous system.

Objective: To examine the prospective effect of cesarean section on pubertal development after controlling the polygenic susceptibility for early puberty in boys and girls,and to provide a reference for choosing a delivervary way beneficial to pubertal development. Methods: Cluster sampling method was used to select 997 students of grade 1 to 3 from 2 priwary schools in Bengbu of Anhui in 2016 to establish the puberty development cohort. Delivery mode,birth weight,gestational age,early teeding way,family income and parental educational background were derived from parental questionnaire. Breast Tanner stage and testicular volume of 997 children were annually assessed. The polygenic risk score (PRS) was computed based on 17 SNPs derived from published genome-wide association studies for early pubertal timing. Kaplan-Meier analyses was used to examine associations between cesarean section with pubertal development in boys and girls. Results: The average age of 997 children was 8.01±0.84 years old, and average BMI was (17.96±2.96) kg/m2. The prevalence of cesarean section for boys and girls was 43.30% and 32.47%, respectively. At baseline, the detection of thelarche in girls delivered by cesarean section (39.89%) was higher than that of girls delivered naturally (4.39%) (χ2=118.65, P<0.01). Similar result was observed during the last follow-up, the detection of thelarche in girls born by cesarean section (95.21%) was higher than girls born vaginally (68.48%), respectively (χ2=51.42, P<0.01). After adjusting for confounding factors (age, BMI, birthweight, infancy feeding, gestational age, household monthly income and parental education), girls delivered by cesarean section had maturation at significantly younger than girls who delivered naturally (Hazard Ratio, HR=1.98，95%CI=1.67-2.34，P<0.01). Conclusion Caesarean section might increase the risk of earlier age at pubertal onset in a sex-specific manner. Whether natural delivery could be one of the protective measures to prevent early pubertal timing in children warrants further investigation.

During an individual's life, puberty is not only a crucial phase for physical development, but a key period for neuroendocrine transformation. As a major neuroendocrine axis, the influence of hypothalamic-pituitary-adrenal (HPA) axis's changes during puberty on mental and physical health, as well as emotional symptoms, is causing a growing attention. However, information of its developing pattern and circadian variation is limited. Recent research has demonstrated that certain developing pattern of HPA axis in puberty is closely related to the adolescent emotional disorders, which highlights the recognition of HPA functions developing paths under both resting and stress state and the evaluation of its prediction effect for the adolescent emotional disorders. Generally, cortisol awakening response is utilised to assess HPA functions. Elaborating the variation of HPA axis functions from the puberty to the later developing process provides much guiding significance for the early screening, intervention and treatment of adolescent emotional disorders.

Objective: To determine the possible relations between hair cortisol concentration (HCC) as a biomarker of chronic stress and childhood overweight and obesity. Methods: Children from grade 1 to 3 from 2 primary schools in Bengbu were invited to participate in the study; and those who suffer from mental disorders, endocrine diseases and those who took hormone drugs were excluded. Parental questionnaire was implemented to collect information on physical activity, screen time, sleep duration, sugar-beverage consumption, as well as socio-economic status. Height and weight were examined. Eligible hair samples from 1 263 children (598 boys and 665 girls) were obtained, and the cortisol content was determined. Multivariate logistic regression model analysis was recruited to examine the association between HCC quartiles and overweight and obesity among boys and girls, respectively. Results: HCC in P₅₀ (P₂₅-P₇₅) among girls and boys was separately 11.86 (10.57-13.15) and 11.71 (10.54-13.09) µg/kg (Z=-0.886, P=0.376); HCC in P₅₀ (P₂₅-P₇₅) among non-overweight, overweight and obesity boys was separately 11.62 (10.45-12.82), 12.24 (10.88-13.55) and 12.30 (10.99-13.53) µg/kg (χ²=8.24, P=0.016); HCC in P₅₀ (P₂₅-P₇₅) among non-overweight, overweight and obesity girls was separately 11.35 (10.06-12.62), 12.07 (11.04-13.21) and 12.59 (11.12-13.63) µg/kg (χ²=36.16, P<0.001). After age, physical activity, screen time, sleep duration, sugared beverages and socio-economic status adjusted, compared with HCC in Q1 level, girls whose HCC in Q2, Q3 Q4 level had higher rates in overweight, with OR (95%CI) at 1.10 (1.04-3.32), 2.59 (1.51-4.45) and 3.16 (1.79-5.57), respectively. Girls with HCC in Q3 and Q4 level had higher rates in obesity, with OR (95%CI) at 2.22 (1.16-4.26) and 5.62 (3.04-10.41). Among boys, highest quartiles in HCC was risk factor for overweight, OR (95%CI) was 2.14 (1.18-3.90). Conclusion Hair cortisol had a significant relation with overweight and obesity in 6-9 years old childhood especially among girls.

Objective: To investigate the longitudinal association between peer bullying victimization and internalizing symptoms, and to examine whether parental warmth buffers against this effect,so as to provide a reference for improving the mental health of the special group of children. Methods: Students in grade 4 and grade 5 from three primary schools in Ma’anshan, Anhui Province were selected in Oct. 2017 with informed consent. Height, weight and pubertal development were objectively assessed. Internalizing symptoms and peer bullying victimization were evaluated at baseline and 1-year follow-up by using the MacArthur Health and Behavior Questionnaire-Child. Children were classified into three groups as non-bullying, non-consistent and consistent bullying victimization group. Multiple linear regression model was used to analyze whether the association between peer bullying victimization and internalizing symptoms varied across different parental warmth groups. Results: Compared with non-bullying victimization group, consistent-and non-consistent bullying victimization had higher internalizing symptoms at baseline and 1-year follow-up(P<0.01). Students reported consistent, non-consistent bullying victimization under moderate parental warmth had lower internalizing symptoms at baseline and 1-year follow-up than those reported under low parental warmth group(P<0.01). Multiple liner regression showed that consistent bullying victimization [β=0.66(95%CI=0.33-0.99), 0.37(95%CI=0.15-0.59), 0.58(95%CI=0.31-0.84), P<0.05] and non-consistent bullying victimization[β=0.33(95%CI=0.18-0.47), 0.28(95%CI=0.13-0.41), 0.29(95%CI=0.08-0.51),P<0.05] were associated with higher 1-year follow-up internalizing symptoms in non-consistent, moderate-and low parental warmth group, while such association was not observed in consistent parental warmth group. Abstract The experience of peer bullying victimization increases the risk of internalizing symptoms and this finding suggests that parental warmth help to buffer children from the internalizing symptoms associated with bullying victimization.

Objective: To explore the impact of adverse childhood experiences on pubertal development,and to provide a reference for conducting the targeted intervention in the early stage. Methods: A cluster sampling method was used to randomly select students of grades 3 to 4 from three primary schools in Ma’anshan municipality of Anhui province in October 2017. Self-reported childhood adverse experiences (ACEs), including physical, emotional and sexual abuse, as well as physical and emotional neglect were collected. Testicular volume and breast Tanner stage were assessed. A follow-up survey was conducted one year later to assess ACEs and pubertal development. ACEs of each dimension were divided into non-exposure group, transient exposure group and sustained exposure group according to ACEs exposure at baseline and follow-up. Multivariate linear regression model was used to analyze the relationship between ACEs exposure and pubertal development after 1-year follow-up. Results: Sustained emotional abuse was positively associated with increased breast Tanner stage and testicular volume (breast: β=0.36, 95%CI=0.09-0.63, P<0.01; testicular volume: β=1.07, 95%CI=0.47-1.66, P<0.01); sustained physical abuse was positively associated with decreased testicular volume (β=-0.83, 95%CI=-1.58--0.08, P<0.05); sustained and transient sexual abuse were significant positively associated with decreased breast Tanner stages (sustained: β=-1.43, 95%CI=-2.86--0.02, P<0.05; transient: β=-0.45, 95%CI=-0.73--0.19, P<0.05); sustained emotional neglect was positively associated with increased breast Tanner staging (β=0.33, 95%CI=0.06-0.61, P<0.05) and decreased testicular volume (β=-0.19, 95%CI=-1.38--0.19, P<0.01). Conclusion Sustained emotional abuse was associated with early pubertal development in boys and girls,and the specitic mechanisam differences of different ACES puberty development effects and the gender differences of the same ACES puberty development effects need to be furthur explored.

Objective: To determine possible relations between early adiposity rebound and adolescent development. Methods: Prospective children cohort from 2 kindergartens selected through clustering convenience sampling method in Anhui Province was established since Sep. 2010. Participants were classified as Cohort 1 (2010), Cohort 2 (2011) and Cohort 3 (2012) according to the recruiting year. Till Sep. 2015, a toal of 802 girls were included in this study, and received follow-up till primary school. During kindergarten period, physical examination was carried out every 3 months, 8 times in total. In primary school, physical examination was carried out annually; till Sep.2015, the cohort 1, 2, 3 took physical examination for 12, 11, 10 times, respectively. Information on household economic status and child physical activity was acquired through parents questionnaire survey, and breast development were assessed through visual inspection and palpation. Adiposity rebound was determined according to Rolland-Cachera's method for each girl. Differences between early adiposity rebound and normal adiposity rebound groups were compared by using t test and χ² test. Multivariate regression analysis was applied to explore the association between early adiposity rebound and breast development. Results: The average age of participants was (8.90±0.87) years old and the BMI was (17.48±2.70) kg/m². The average age at adiposity rebound was (6.16±0.90) years old and the BMI was (15.33±1.82) kg/m². Premature breast development was found significantly higher in girls in early adiposity rebound group (27.8%, 54/802) than it in normal adiposity rebound group (13.7%) (P<0.001). After current adiposity, age, household economic status, childhood physical activity adjusted, the OR of premature beast development in early adiposity rebound group was 2.41(95%CI: 1.41-4.12). Conclusion Early adiposity rebound increases the risk of premature puberty in girls.

Objective:To examine the prospective association of pubertal timing and tempo with depressive symptoms in adolescents.Methods:Since 2013, 2 084 students in grade 1-3 were selected from two primary schools in Bengbu, Anhui Province were selected by using convenience sampling method to establish the adolescence pubertal development cohort. Followed up for 6 years, physical examination, secondary sexual development evaluation (testicular volume for boys and breast development for girls) and depressive symptoms were evaluated biennially. Non-linear growth model was used to estimate pubertal timing and tempo for boys and girls respectively. Depressive symptoms were interviewed by using the Short Mood & Feeling Questionnaire (SMFQ) at baseline and Mood & Feeling Questionnaire (MFQ) during follow-up for students in grade 1-2. Children Depression Inventory (CDI) was used for students in grade 3 at baseline and during follow-up. Depressive symptom scores were standardized by using the Z-score method. Multivariate linear regression model was used to analyze the predictive effects of modeling pubertal timing and tempo on depressive symptoms of adolescence boys and girls. Results:There were 1 909 students with complete questionnaire and puberty development information, including 1 052 boys (59.19%) and 857 girls (43.81%), with average age about (13.94±0.87) years and 91.60 percent follow-up rate. The average modeling pubertal timing of girls (11.25 years) was earlier than that of boys (12.70 years), and the average pubertal tempo of girls about 1.47 Tanner stage/year was faster than that of boys about 1.28 Tanner stage/year. After controlling for depressive symptoms, maternal education and adverse childhood experiences at baseline and age, body mass index (BMI) classification and sleep time during follow-up, this predictive effect of pubertal timing and tempo on depressive symptoms was only significant among girls. Compared with girls with on time pubertal timing, girls in the delay timing group had a lower level of depressive symptoms (β=-0.19, 95% CI:-0.34,-0.01). Compared with girls in average pubertal tempo group, the fast tempo group associated with an increasing risk of depressive symptoms (β =0.23, 95% CI: 0.05, 0.40), while the slow tempo group associated with an decreasing risk of depressive symptoms (β =-0.21, 95％ CI:-0.39,-0.03). Insignificant effects were found in puberty timing and tempo on depressive symptoms of boys ( P>0.05). Conclusion:Fast pubertal tempo increases the risk of development of depressive symptoms of adolescent girls. There is no predictive effect of pubertal timing and tempo on depression symptoms of adolescent boys.

Objective:To examine the prospective association of pubertal timing and tempo with depressive symptoms in adolescents.Methods:Since 2013, 2 084 students in grade 1-3 were selected from two primary schools in Bengbu, Anhui Province were selected by using convenience sampling method to establish the adolescence pubertal development cohort. Followed up for 6 years, physical examination, secondary sexual development evaluation (testicular volume for boys and breast development for girls) and depressive symptoms were evaluated biennially. Non-linear growth model was used to estimate pubertal timing and tempo for boys and girls respectively. Depressive symptoms were interviewed by using the Short Mood & Feeling Questionnaire (SMFQ) at baseline and Mood & Feeling Questionnaire (MFQ) during follow-up for students in grade 1-2. Children Depression Inventory (CDI) was used for students in grade 3 at baseline and during follow-up. Depressive symptom scores were standardized by using the Z-score method. Multivariate linear regression model was used to analyze the predictive effects of modeling pubertal timing and tempo on depressive symptoms of adolescence boys and girls. Results:There were 1 909 students with complete questionnaire and puberty development information, including 1 052 boys (59.19%) and 857 girls (43.81%), with average age about (13.94±0.87) years and 91.60 percent follow-up rate. The average modeling pubertal timing of girls (11.25 years) was earlier than that of boys (12.70 years), and the average pubertal tempo of girls about 1.47 Tanner stage/year was faster than that of boys about 1.28 Tanner stage/year. After controlling for depressive symptoms, maternal education and adverse childhood experiences at baseline and age, body mass index (BMI) classification and sleep time during follow-up, this predictive effect of pubertal timing and tempo on depressive symptoms was only significant among girls. Compared with girls with on time pubertal timing, girls in the delay timing group had a lower level of depressive symptoms (β=-0.19, 95% CI:-0.34,-0.01). Compared with girls in average pubertal tempo group, the fast tempo group associated with an increasing risk of depressive symptoms (β =0.23, 95% CI: 0.05, 0.40), while the slow tempo group associated with an decreasing risk of depressive symptoms (β =-0.21, 95％ CI:-0.39,-0.03). Insignificant effects were found in puberty timing and tempo on depressive symptoms of boys ( P>0.05). Conclusion:Fast pubertal tempo increases the risk of development of depressive symptoms of adolescent girls. There is no predictive effect of pubertal timing and tempo on depression symptoms of adolescent boys.