Role of miR-146a in ultraviolet A-induced photoaging of human skin fibroblasts and its mechanism / 中华皮肤科杂志

Objective To investigate miR-146a-Smad4 expression during ultraviolet A(UVA)-induced photoaging of human skin fibroblasts (HSFs), and to evaluate effects of up-regulation of miR-146a expression on its target gene Smad4 and cell photoaging. Methods HSFs were isolated from the prepuce, and subjected to primary culture and maintained up to 10th passage. Then, the HSFs were classified into 4 groups: blank control group receiving no treatment, UVA group irradiated with 10 J/cm2 UVA, miR-146a group transfected with a lentiviral vector expressing miR-146a, UVA+ miR-146a group transfected with the lentiviral vector expressing miR-146a followed by UVA radiation. Real time PCR was performed to measure miR-146a expression in HSFs in the UVA group on day 0, 3, 7 and 14 after UVA radiation.Fluorescence microscopy was carried out to estimate transfection efficiency on day 7 and 14 in the miR-146a group after transfection, and real time PCR was performed to quantify miR-146a expression in these cells. Methyl thiazolyl tetrazolium (MTT)assay was conducted to evaluate proliferative activity of HSFs, real time PCR to quantify mRNA expressions of photoaging-related genes p53, p21 and p16, and Western blot analysis to measure Smad4 protein expression in these cells. Statistical analysis was carried out by using repeated-measures analysis of variance and factorial design analysis of variance. Results Repeated-measures analysis of variance showed that the expression of miR-146a decreased over time in both the UVA group and blank control group(F = 213.840, P 0.05). Factorial design analysis of variance showed that UVA radiation had an inhibitory effect on the proliferative activity of HSFs (P 0.05). Real time PCR and Western blot analysis both revealed that UVA radiation could increase the expressions of p53, p21 and p16 mRNAs as well as Smad4 protein(all P 0.05). Conclusion The expression of miR-146a is inhibited in UVA-induced photoaged HSFs, and its up-regulation may counteract cell photoaging by suppressing Smad4 expression in, and promoting proliferation of, photoaged HSFs.

Status analysis on child neglect of rural pupils aged 6-11 year-old in China / 中华预防医学杂志

<p><b>OBJECTIVE</b>To analyze pupils' neglect status aged 6-11 years in China's rural.</p><p><b>METHODS</b>According to the principle of multi-stage stratified cluster sampling, to extract 7,943 pupils aged 6-11 years for the survey from 56 primary schools in 28 counties in nine provinces, from December 2012 to March 2013. Proceed questionnaire survey by the scale from "Neglect Evaluation Norms for pupils aged 6-8 years in Rural Areas of China" and "Neglect Evaluation Norms for pupils aged 9-11 years in Rural Areas of China". And analyze neglect rates and neglect degrees of the different grades, gender, family types and different levels of neglect.</p><p><b>RESULTS</b>The total neglect rate of China's rural pupils aged 6-8 and 9-11 years was 40.2% (1,258/3,130) and 42.5% (1,498/3,526) respectively, which differences had no statistical significant (χ2=3.59, P=0.058); the total neglect degree was 46.04±8.87 and 44.78±10.43 respectively, which differences had statistical significant (t=5.01, P<0.001); the differences of neglect rates and neglect degrees between male (41.4% (657/1,587) and 46.28±8.76) and female (39.0% (601/1,543) and 45.78±8.97) of aged 6-8 were all no statistical significant (χ2=1.87, P=0.171; t=1.49, P=0.136); the neglect rates and neglect degrees of male (46.3% (816/1,763) and 45.53±10.11) were higher than female (38.7% (682/1,763) and 44.06±10.69) in the group of aged 9-11, which differences were all statistical significant (χ2=20.84, t=3.97, P<0.001); the male neglect rate in the group of aged 6-8 in social neglect (11.7% (198/1,691)) and the neglect degree in educational neglect (48.09±9.70) were higher than female (9.4% (155/1,648) and 47.37±9.89), which differences were all statistical significance (χ2=14.55, P<0.001, t=2.22, P=0.026), the male neglect rate in the group of aged 9-11 in physical neglect (20.4% (398/1,954)) was higher than female (16.7% (326/1 957)), which differences had statistical significance (χ2=8.92, P=0.003).</p><p><b>CONCLUSION</b>The neglect status of Chinese pupils aged 6-11 years in rural was serious, and we should find out risk factors and provide efficient prevention measures.</p>

Impact factors analysis on child neglect of children aged 3-6 year-old in rural areas of China / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the major influence factors of child neglect among rural areas children aged 3-6 years in China.</p><p><b>METHODS</b>According to multi-stage stratified cluster sampling, distribution characteristics and the level of economic development, we randomly sampled 10 provinces, 26 cities (8 capital cities, 16 prefecture-level cities and 2 municipalities) using lottery method. Depending on the different level of economic and cultural, we sampled one medium county from each city (municipalities sampled two counties), a total of 28 counties. All towns were divided into high, medium and low three levels in accordance with economic and cultural, each level sampled one town, each conty sampled three towns, a total of 84 towns were sampled. Each town sampled 40 children, including 20 boys and 20 girls (including each 10 children aged 3 to 6, half and half boys and girls). 4 096 rural children aged 3-6 years old were sampled and surveyed of China. Based on "the Neglect Norms for Children Aged 3 to 6 Years Old in Rural Areas of China" explored the risk factors of child neglect. SPSS 18. 0 for windows was employed for statistics analysis. Multifactorial analysis was conducted through multivariate Logistic Regression.</p><p><b>RESULTS</b>The total neglected rate of the rural children aged 3 to 6 was 53.7% (2,047/3,810), and the total neglected degree was 44.42±7.57. The multivariate logistic regression analysis showed that, neglected risk of the boys was higher than the girls (OR=1.30, 95% CI: 1.13-1.49), neglected risk of children were higher, when their mothers were migrant workers (OR=2.18, 95% CI: 1.59-3.00); the lower educational level of mothers, the higher neglected risk of children, the children in families that mothers only received primary education or was uneducated were most likely to be neglected (OR=2.92, 95% CI: 1.91-4.47), the second was the families that mothers received primary education (OR=1.86, 95% CI: 1.29-2.69); the lower the household income, the easier to be neglected the children, the easiest one was the household income less than 5000 RMB (OR=2.85, 95% CI: 2.14-3.79), the second was 5000-9,999 RMB (OR=1.76, 95% CI: 1.40-2.20); the relationship between children and mothers was distanced, neglect risk of children was higher (OR=3.88, 95% CI: 1.31-11.52); left behind children were easier to be neglected (OR=1.30, 95% CI: 1.09-1.54).</p><p><b>CONCLUSION</b>The influence factors among children aged 3-6 years in the rural areas of China were boys, mother's occupation, maternal education level, the relationship between children and mothers, household income and left-behind children.</p>

Expression and clinical significance of miR-34b in children with acute leukemia / 中华实用儿科临床杂志

Objective To research the expression,methylated regulation and clinical significances of miR-34b in chidren with acute leukemia(AL).Methods The methylation status of miR-34b promoter CpG islands were detected with methylation-specific polymerase chain reaction (MSP) in patients with AL.Then the expression of miR-34b was compared,which was detected by Taqman real-time fluorescence quantitative polymerase chain reaction (qRT-PCR),between the AL patients and normal group,in order to analyze their relationship with the clinical indicators.Resuits In 8 leukemia cell lines (U937,HL-60,MV4-11,M2R,K562,Raji,CCRF,DAMI) showed methylation,the positive rate of the methyl was 100％.Thirty-one acute lymphocytic leukemia(ALL) pediatric patients were newly diagnosed,and 24 cases showed methylation,the methylation positive rate 77.42％ (24/31).Nineteen acute myeloid leukemia(AML) patients were newly diagnosed,and 8 cases showed methylation,the methylation positive rate was 42.11％ (8/19 cases).There was no methylation in the 23 cases of normal children.The relative expression levels of miR-34b in the normal group,the group of leukemia cell lines,the group of ALL pediatric patients with newly diagnosed,the AML group and the group with mixed lineage leukemia gene rearrangement MLL+ were 5.22 ± 1.15,0.03 ± 0.03,1.65 ± 0.69,0.18 ± 0.06,0.64 ± 0.34,respectively.The findings indicated that there were significant differences in the relative expression levels of miR-34b between the normal group and the group of leukemia cell lines,the ALL group,the AML group,and the MLL+ group.The relative expression and methylated level of miR-34b had no statistically difference in gender,age at diagnosis,WBC count,chromosome,fusion gene,MLL gene rearrangement,and the minimal residual disease(LDH) levels in newly diagnosed AML patients(all P ＞ 0.05).And the relative expression and methylated level of miR-34b had no statistically difference in gender,age at diagnosis,WBC count,immunophenotype,chromosome,fusion gene,MLL gene rearrangement,TEL/AML1 gene,risk stratification,the minimal residual disease (MRD) in thirty-three days and the LDH levels in newly diagnosed ALL patients(all P ＞ 0.05).But as for the response to prednisone experiment,there was a significant difference between the sensitive group and the non sensitive group(P ＜ 0.05).Conclusions The expression level of miR-34b in AL was significantly lower and it was regulated by methylation mechanism,which implies that miR-34b may play a role of a tumor suppressor gene in the pathogenesis of leukemia.MiR-34b may affect the early treatment response of ALL patients,and it may be an indicator of risk stratification and poor prognosis in pediatric ALL.

Clinical analysis of 129 children with lead poisoning / 中华实用儿科临床杂志

Objective To assess the distribution of blood lead levels and the sources of lead exposure in children with lead poisoning,and thus to offer recommendations for clinical diagnosis and treatment of childhood lead poisoning.Methods The clinical data of 129 patients with lead poisoning was collected and analyzed at the Out-patient Department of Lead Poisoning Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine during Sep.2012 and Aug.2013 were collected and analyzed.All children were required to fill out theoutpatient questionnaire on heavy metal (including children's demographic data,growth assessment,frequency of hand-mouth behavior,and the behavior of washing hands before eating,dietary,sources of lead exposure,and the family environment,etc.).Other data of medical history,physical examinations (height,weight,growth and development) were also collected.Blood samples were collected to measure blood lead level by graphite furnace atomic absorption spectrometry.Results (1) The above 129 patients were from 14 provinces (municipalities and autonomous regions),including 64 cases in Zhejiang (49.6％),30 cases in Shanghai (23.0％),13 cases in Jiangsu (10.1％),6 cases in Jiangxi (4.7％),5 cases in Hebei (3.9％),2 cases in Anhui and Guangdong (1.6％) ;and 1 case in Fujian,Henan,Hunan,Jilin,Inner Mongolia,Heilongjiang and Shandong (0.8％),respectively.(2) In the patients,the blood lead level was 17.0-892.0 μg/L[(251.5 ±155.8) μg/L] and the median was 235.0 μg/L.(3)The mean age of the children was 4.3 years.Fifteen cases were less than 1 year old,and the mean blood lead level was (367.8 ± 137.7) μg/L.Thirty-seven cases were 1-3 years old children,and the mean blood lead level was (250.5 ± 116.3) μg/L.Fifty cases were 3-6 years old children,and the mean blood lead level was (237.7 ± 179.7) μg/L.Twenty-seven cases were over 6 years old,and the mean blood lead level was (213.9 ± 141.8) μg/L.(4) One hundred and eleven cases of the children could find the definite sources of lead exposure,mainly from industrial pollution (35.7％) and domestic pollution (64.3％).The blood lead levels in 18 cases were less than 100 μg/L,and their definite lead pollution source was not found.(5) Most of the children had the symptoms of inattention,hyperactivity,aggressive behavior,constipation and abdominal pain,and so on.(6)Logistic regression analysis of children with blood lead levels ≥ 235.0 μg/L showed that lead pollutants and age were the main risk factors for lead poisoning.Conclusions Industrial pollution are associated with higher blood lead levels among children aged 0-6 years old (occupying one-third of the pollutants).The younger children tend to have higher blood lead levels,and the data also suggest that greater attention should be paid to children who used red powder.

Development of Neglect Evaluation Scale for primary school students aged 6-11 years old in rural areas of China / 中华预防医学杂志

<p><b>OBJECTIVE</b>To develop a standard Neglect Evaluation Scale which is suitable for rural primary school students in China.</p><p><b>METHODS</b>According to the principle of multi-stage stratified cluster sampling, a field investigation was conducted among primary school students in 28 counties from 7 provinces and 2 municipalities. The questionnaires were self-designed, and determined by multiple rounds of pilots and revisions. Among 5 862 students, 2 792 (6-8 years' old) and 3 070 (9-11 years' old) were investigated by using two kinds of Neglect Evaluation questionnaires, respectively. After project analysis, factor analysis, reliability and validity analysis, the reliability and stability of the scale were tested. Then percentile method was used to determine the evaluation standard to develop and finalize the formal scale.</p><p><b>RESULTS</b>The numbers of the remaining items for 6-8 year-old group and 9-11 year-old group is 69 and 58, respectively, both of which contain six levels of neglect, including body, emotion, health care, education, security, and society. 4 times of factor analysis were conducted in both of the two groups. The factor loadings in these two groups were 0.290-0.700 and 0.276-0.729 respectively. Reliability test results showed that the two kinds of scales' Cronbach alpha coefficient were 0.924 and 0.929 respectively, split-half reliability were 0.891 and 0.904 respectively, the retest reliability were 0.559 and 0.892 respectively, the differences were statistically significant (P < 0.05). The retest reliability among 6-8 year-old group in medical neglect level had no statistical significance, but was close to the cut-off point (P = 0.054). The test results of external validity indicated that both of the two scales could reflect the neglect status of the subjects (P < 0.05), but the subjective and objective evaluation towards neglect were not consistent. The cut-off points for judging whether the children were neglected or not among 6-8 year-old and 9-11 year-old groups were 159 and 137 respectively; and the adjusted values were 160 and 135 respectively.</p><p><b>CONCLUSION</b>The two norm scales developed by this study showed good discriminability, reliability, validity, and stability. The norms developed on the basis of the scales was suitable for the situation of rural students in primary schools in China.</p>

Effect of DNA methylation on the proliferation of leukemia cells regulated by miR-34b / 中华儿科杂志

<p><b>OBJECTIVE</b>To study the expression level and CpG island methylation status of miR-34b in leukemia cell lines and to research the effect of DNA methylation on the proliferation of leukemia cells regulated by miR-34b.</p><p><b>METHOD</b>Taqman real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was carried out to detect the relative expression of miR-34b in control group (bone marrow cells of 20 children without blood disease) and 8 leukemia cell lines (U937, HL-60, MV4-11, M2R, K562, Raji, CCRF, DAMI). Methylation-specific polymerase chain reaction (MSP) was carried out to detect the methylation differences of miR-34b in control group (Bone marrow cells of 23 children without blood disease), 8 leukemia cell lines. HL-60 and K562 were treated with methyltransferase inhibitor (5-aza-2-dC) for further detection of its methylation status and expression of miR-34b. Hsa-miR-34b mimics was transfected into K562 cell by liposome, the transfection efficiency was detected by flow cytometry. The cell proliferation of hsa-miR-34b transfected group in each stage was measured with CCK-8 assay, and then compared with non-transfected group and negative control group.</p><p><b>RESULT</b>The relative expression level of miR-34b in the group of children without blood disease and the group of leukemia cell lines were 5.22 ± 1.15, 0.03 ± 0.03. The results showed that, the group of leukemia cell lines was significantly different from the control group (t = 4.538, P < 0.01) . Eight leukemia cell lines showed methylation, the positive rate of the methyl was 100%. There was no methylation in the 23 cases of control group. After leukemia cell lines HL-60 and K562 were treated with 5-aza-2-dC, the methylated bands became obviously weakened, and the relative expression levels of miR-34b substantially increased 49.5 times and 18.8 times respectively. After hsa-miR-34b mimics was transfected into K562 cell by liposome, its transfection efficiency detected by flow cytometry was 61% and the cell proliferation was measured with CCK-8 assay from which it was found that the cell proliferation was significantly suppressed compared with the control group at 48 h (t = 9.303, P < 0.01), 72 h (t = 65.617, P < 0.01), 96 h (t = 36.878, P < 0.01) and 120 h (t = 18.748, P < 0.01) in hsa-miR-34b transfected group, with the inhibition rate of 12.2% (48 h), 45.7% (72 h), 32.5% (96 h) and 22.9% (120 h).</p><p><b>CONCLUSION</b>The hypermethylation of promoter leads to decrease in the expression levels of miR-34b in leukemia cell lines, which attenuate mechanism of proliferative inhibition may be one of the reasons of occurrence or development of childhood leukemia.</p>

Environmental factors associated with developmental coordination disorder in preschool children in urban area of Suzhou city / 中华儿科杂志

<p><b>OBJECTIVE</b>A population-based study on developmental coordination disorder (DCD) was conducted in Suzhou to explore the impacts of family and kindergarten environment on pre school children with DCD so as to provide a basis for etiological research and early intervention.</p><p><b>METHOD</b>Stratified clustered sampling was used to select 160 classes from randomly selected 15 public nursery schools distributed throughout the five main districts in Suzhou city. A total of 4 001 children were included in the study. The family environment scale on motor development for urban preschool children (FESMDPU) which was established by our study group and early childhood environment rating scale-revised (ECERS-R) which has been applied well in China were used to assess the family and kindergarten's environment. The multilevel logistic regression was used to analyze the risk factors of DCD when kindergarten environment were considered as "context variables" and the family environment as "individual variables".</p><p><b>RESULT</b>According to DSM-IV criteria, a total of 330 children were diagnosed as DCD. The prevalence of DCD was 8.3%. However, there were differences between the two groups in age, gender and Kaup index (all P < 0.05). The results of ECERS-R and FESMDPU showed that the scores of "class space and faculty" "class activity" "class interaction" "family material environment" "family rearing environment" in DCD group were 48.00, 51.00, 49.00, 39.00, and 30.00, respectively, which were higher than those of control group (45.00, 50.00, 47.00, 41.00, 31.00) with statistical significance (U = 455 446.000, 550 787.000, 508 109.000, 543 159.000, and 490 119.000, P < 0.05 for all comparisons) . The score of ECERS-R and FESMDPU were grouped into different levels using the method of K-MEANS. The school and family environment were compared after the clustering. The results showed that the distribution of the rates in different levels between the DCD and control group were different with statistical significance (χ(2) = 51.091, 9.295, 35.464, 15.174, 13.500, P < 0.05 for all comparisons) . There was no significant difference between DCD and non-DCD children in parents' schooling years and family per-capita income of every month (all P > 0.05). The results of the multilevel logistic regression model showed that when children's gender, age and Kaup index were controlled, "class space and faculty" "class activity" "class interaction" "family material environment" "family rearing environment" were included in the model (OR was 3.486, 1.840, 1.623, 1.531, and 1.379, respectively, P < 0.05 for all) .</p><p><b>CONCLUSION</b>The prevalence of DCD in study area was higher than that reported by European and American countries. The family and kindergarten environment may affect the incidence of DCD in preschool children. Parents and preschool educational and child health care practitioners should conduct the early prevention and intervention on DCD based on the risk factors of environment in preschool children.</p>

Prenatal and perinatal risk factors for developmental coordination disorder in children / 中华流行病学杂志

Objective To explore the prenatal and perinatal risk factors for developmental coordination disorder (DCD) in children so as to provide a basis for etiological studies and early diagnosis on DCD.Methods Stratified clustered sampling was used to select 160 classes from randomly selected 15 public nursery schools which were distributed throughout the five main districts in Suzhou city.A total of 4 001 children were included in the study.Multilevel logistic regression and structural equation model (SEM) were used to analyze the risk factors of DCD.Results Results from the multilevel logistic regression model showed that when factors as:class internal clustering,children' s gender,age and Kaup value etc.,were under control,factors as acute fetal distress,premature and newborn pathological jaundice would be included in the model (OR values were 7.030,3.374,1.993,with all their Ps＜0.001),and showing these variables were related to DCD.Results of SEM showed that the related coefficient between ‘Minimal Brain Dysfunction' and ‘Motor coordination skill' was-0.18 with statistical significance (P＜0.05).It also indicated that the minimal brain dysfunction would negatively impact on the children' s motor skill.Conclusion There were risk factors of DCD noticed during the prenatal and peri-natal periods.Practitioners working at the maternity and child health care areas should improve the skills in assessing and monitoring the prenatal and peri-natal risk factors related to DCD,and to better carry out early screening,diagnosis,treatment and prevention of DCD.

An investigation in 370 cases of child neglect / 中国组织工程研究

BACKGROUND: Child neglect is one of the most important aspects of child abuse and has become a serious problem influencing children's development all over the world. Therefore, it is necessary to nderstand the current conditions and influential factors of child neglect and establish effective evaluation methods in China.OBJECTIVE: To investigate the current conditions and influential factors of urban child neglect in Suzhou city.DESIGN: A comparative analysis of sampling survey.SETTING: Department of Children's Health Care, Soochow University Affiliated Children's Hospital.PARTICIPANTS: This study was conducted in one public kindergarten and two private kindergartens of urban areas in Suzhou city between March and October 2003. Totally 370 children (196 boys and 174 girls) aged between 4 to 6 years old without obvious physical and mental diseases were voluntarily investigated, and they had been living with the person that answered the questionnaire regarding to child neglect in China in the recent 1 year. METHODS: Questionnaire regarding to child neglect in China was sent to each volunteer.he questionnaire evaluates the current conditions of child neglect by 20 items of 5 aspects including physical care (buy fruit for child, consider the nutritional need of child, care about the body height and body mass of child), emotional care (chat with child, play and tour with child, encourage child to do the things whatever they can, listen to what the child feels, introduce the child to the guests when they come to your home, let him or her stop immediately when child cries, care about the activities in kindergarten), education care (buy books for child, tell stories to child, recognize and correct the bad behavior of child, answer the questions asked by child seriously), safety care (protections against fire and electricity at home, warn child against matches, knives and other sharp things, teach child how to cross the road) and health care (prepare the common medicine for child at home, urge child to brush teeth, correct the bad posture). The volunteers were asked to answer each question objectively according to the conditions of child within the recent 1 year, and each question should be answered by selecting only one choice (never, occasionally, usually, always), and anyone should not be omitted. Meanwhile,the educational background and occupation of the parents were also investigated.MAIN OUTCOME MEASURES: The general status of child neglect and the effects of different family educational background and occupation of parents on child neglect were observed. RESULTS: Totally 370 questionnaires were sent out, and 370 qualified ones were recollected, the effective rate was 100%. ① In Suzhou city, the children were given better regards and good care in the aspects of education, medicine and safety, but neglects on children's physical and mental care still exists in a few families. ② Families of different educational background could think highly of the medical care, safety and psychical care of children, but neglects on education and emotional development more commonly existed in the families with the educational background under university. ③ All the families with different occupation could think highly of the medical care, safety and physical care of the children, but education and emotional development were better in the cadre families than in the ordinary families.CONCLUSION: In Suzhou city, the children are given better regards and good care in the aspects of education, medicine and safety, but some bad manifestations of child neglect exist in education and emotion, which are associated with the educational background and occupation of the parents.Further studies about the evaluation, prevention and intervention are needed for child neglect in China.