Research progress in kidney dendritic cells / 浙江大学学报·医学版

Kidney dendritic cells(DC) play important roles in the pathogenesis of kidney diseases. Kidney DC presents anti-inflammatory effects in certain kidney diseases, sometimes presents pro-inflammation in other diseases, and sometimes their effects are changing in different stages of the disease, suggesting that the differentiation and function of kidney DC may be influenced by microenvironment. This article reviews the origin and distribution of kidney DC subsets and their roles in the pathogenesis of kidney diseases such as lupus nephritis and pyelonephritis, and the functional regulation of kidney DC by proximal tubule epithelial cells.

Progress in TcpC research / 浙江大学学报·医学版

TcpC is a homolog of the Toll/interleukin-1 receptor (TIR) domain and is secreted by uropathogenic E. coli strain CFT073. TcpC can bind to MyD88, hereby exerting inhibitory effects on macrophages. TcpC represents an important virulence factor that promotes bacterial survival and pathogenicity. TcpC plays a critical role in urinary tract infection, particularly in the pathogenesis of pyelonephritis. In this review,the progress and prospects in TcpC research are discussed.

TcpC induces apoptosis of macrophages through promoting ROS production / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of Toll/interleukin 1 receptor domain-containing protein(TcpC)on macrophages and its mechanisms.</p><p><b>METHODS</b>Murine macrophage J774A cells were co-cultured with TcpC producing wild type E. coli strain CFT073 (TcpC(wt)) or tcpc gene-deleted CFT073 mutant (TcpC(mut)) in Transwell system, respectively. Apoptosis of J774A cells co-cultured with TcpC(wt) or TcpC(mut) was analyzed by Annexin/PI double staining. The levels of reactive oxygen species (ROS) in J774A cells were determined by DCFH-DA staining after treatment with TcpC(wt) or TcpC(mut) at 6 h, 12 h,24 h or 36 h. After the ROS was scavenged by N-acetylcysteine (NAC), the changes of J774A cell apoptosis were also examined. The expression of caspase-3 in J774A cells co-cultured with TcpC(wt) or TcpC(mut) in the presence or absence of 0.1 mmol NAC was detected by Western blot.</p><p><b>RESULTS</b>J774A cells co-cultured with TcpC(wt) for 24 h or 36 h showed significantly increased apoptosis (27.39% ± 4.05% and 28.45% ± 4.55%,respectively) when compared to control group (7.96% ± 1.63% and 10.55% ± 1.44%,P<0.01) or TcpC(mut) group (11.45% ± 2.77% and 19.26%± 2.89%,P<0.01). Levels of ROS in J774A cells treated with TcpC(wt) for 24 h (108.8 ± 9.73) or 36 h (100.3 ± 10.11) were significantly higher than those in control group (56.8 ± 4.11 and 52.8 ± 4.42,P<0.01) or TcpC(mut) (69.7 ± 5.66 and 62.6 ± 4.56, P < 0.01). The pro-apoptotic effects of TcpC(wt) on J774A cells were reversed by 0.1 or 1 mMol NAC treatment. Expression of caspase-3 in J774A cells co-cultured with TcpC(wt) (0.43 ± 0.04) decreased significantly when compared to control group (0.75 ± 0.08,P<0.05) or TcpC(mut) group (0.80 ± 0.12,P<0.05). However,total caspase-3 expression was restored in J774A cells co-cultured with TcpC(wt) in the presence of 0.1 mmol NAC (0.80 ± 0.09).</p><p><b>CONCLUSION</b>TcpC can promote ROS production in macrophages,hereby inducing macrophage apoptosis.</p>

TcpC induces apoptosis of human vascular endothelial cells and its mechanisms / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of TcpC on human umbilical vascular endothelial cells (HUVECs) and its mechanisms.</p><p><b>METHODS</b>HUVECs were co-cultured with TcpC secreting wild-type E. coli strain CFT073 (TcpC(wt)) or tcpc gene-deleted CFT073 mutant strain (TcpC(mut)) in transwell system,respectively. Apoptosis of HUVECs was analyzed by Annexin-V/PI double staining. Mitochondrial membrane depolarization was detected by JC-1 staining. Expression of apoptosis-related proteins in HUVECs was determined by Western blot.</p><p><b>RESULTS</b>HUVECs showed morphological changes after co-cultured with TcpC(wt) for 24 h: the cells became detached and cell debris increased,and cell number was also decreased when compared to HUVECs co-cultured with TcpC(mut). The apoptosis of HUVEC cells co-cultured with TcpC(wt) for 24 h significantly increased,compared to that of control group and TcpC(mut) group (60.1% 9.7% compared with 9.0% 1.3% and 16.9% 0.4%,respectively, P<0.05); meanwhile the mitochondrial depolarization of HUVECs co-cultured with TcpC(wt) was significantly increased,compared to that in control and TcpC(mut) groups (64.5% 0.9% compared with 14.5% 2.1% and 15.6% 3.3%, respectively,P<0.05). Cleavage of PARP and inhibition of Mcl-1 and XIAP expression were seen in HUVECs co-cultured with TcpC(wt),but not in groups of control and TcpC(mut).</p><p><b>CONCLUSION</b>TcpC secreted from CFT073 can induce apoptosis of HUVECs through mitochondrial pathway, in which PARP is cleaved and Mcl-1 and XIAP expressions are inhibited.</p>

Bone alkaline phosphatase and N-MID osteocalcin in monitoring of osteoporosis treatment with recombinant human parathyroid hormone 1-34 / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To evaluate the application of bone turnover markers bone alkaline phosphatase (BALP) and N-MID osteocalcin (N-MID) in monitoring of osteoporosis treatment with recombined parathyroid hormone 1-34 (rhPTH1-34).</p><p><b>METHODS</b>The bone mineral density (BMD) of the lumbar spine L2-L4 and the proximal femur were examined by dual energy X-ray absorptiometry (DXA) before and 6 and 12 months after rhPTH 1-34 treatment. Meanwhile, serum levels of BALP and N-MID were detected by electro-chemiluminescence assay.</p><p><b>RESULTS</b>Six months after rhPTH 1-34 treatment, the BMD of proximal femur remained unchanged, and the BMD of the lumbar L2-L4 spine increased from 0.753 g/cm(2) to 0.781 g/cm(2) (P<0.05); while serum levels of N-MID increased from 15.46 ng/ml to 27.07 ng/ml(P<0.01), BALP from 14.05 μg/ml to 24.31 μg/ml(P<0.01). Twelve months after drug administration, no significant changes were observed in BMD of proximal femur, and the BMD of the lumbar spine L2-L4 increased from 0.753 g/cm(2) to 0.807 g/cm(2)(P<0.01) while serum levels of N-MID and BALP increased from 15.46 ng/ml and 14.05μg/ml to 49.38 ng/ml and 33.99 μg/ml, respectively (both P<0.01).</p><p><b>CONCLUSION</b>Serum levels of BALP and N-MID are more sensitive than BMD. Combination of two methods may provide better indicators for monitoring of osteoporosis treatment.</p>

Development of national neglect norm for urban primary school students of China / 中华预防医学杂志

<p><b>OBJECTIVE</b>To develop the national neglect norms for urban primary school students in China.</p><p><b>METHODS</b>According to multi-stage stratified cluster sampling principle, 24 cities of 13 provinces (municipalities) in China were selected during December 1 to 31, 2008. A total of 1491 students in grade 1 - 3 and 2236 students in grade 4 - 6 were selected. Questionnaire was designed by authors and the final norms were determined through several statistical analysis methods, such as item analysis method, factor analysis method, reliability analysis method. The reliability analysis and validity analysis were used to test the stability and reliability of the norms. The evaluation criteria of the scale was determined by the percentile method, then the initial development of the norm was completed.</p><p><b>RESULTS</b>The two questionnaires of grade 1 - 3 and grade 4 - 6 students consisted of 55 and 57 items, respectively, whose item loadings were ranged from 0.301 to 0.687 and 0.321 to 0.730, which met the statistical requirements. For grade 1 - 3 students, the scale's total Cronbach α coefficients was 0.914, the total split-half reliability coefficients was 0.896, the Cronbach α coefficients of four level was above 0.737 except medical and social neglect, split-half reliability was ranged from 0.461 to 0.757; for grade 4-6 students, the scale's total Cronbach α coefficients was 0.916, split-half reliability was 0.883, except social neglect, the Cronbach α coefficients of other level was ranged 0.457 to 0.856, split-half reliability was ranged from 0.500 to 0.798. The total neglect cut-off score of the two scales grade 1-3 and 4-6 were 125 and 155, respectively.</p><p><b>CONCLUSION</b>The structure of two norms was reasonable. The scales have good stability and reliability.</p>

Development of the National Norms of Negligence on children aged 0 to 35 months in rural areas of China / 中华流行病学杂志

Objective To develop the National Norms of Negligence (NNN) for rural children aged 0 to 35 months.Methods According to multi-stage stratified cluster sampling principle,10 provinces or municipalities (Jilin,Shaanxi,Shanxi,Beijing,Anhui,Jiangsu,Hunan,Hubei,Yunnan,Chongqing) in China were selected.A national research group was formed collaboratively.A questionnaire was designed by ourselves.According to several statistical analysis methods,such as item,factor and reliability analysis etc.we determined the norm.The evaluation criteria of the scale were determined by percentile method.Finally,the reliability and validity of the norm were evaluated.Results In total,2310 children were surveyed,in which the effective sample were 2227,with an effective rate as 96.4％.The scale consisted of 6 neglected dimensions and 65 items in total.The total Cronbach's α coefficient of the scale was 0.903,with the split-half reliability coefficient as 0.829,the parallel reliability as 0.720 and the re-test reliability as 0.678,respectively.The total neglect cut-off score of this scale was 139.Conclusion The scale seemed to have perfect stability and reliability and all the statistical indicators met the psychometric demands.

Comparative study on the situation of neglected children aged 3-6 year-olds between urban and rural areas of China / 中华流行病学杂志

Objective To investigate and analyze the situation of urban and rural neglected children aged 3-6,in China,so as to provide basis for the analysis and comparison on relevant risk factors.Methods 1163 urban children aged 3-6 (with 49.6％ males and 4.5％ with minority ethnicity) were investigated from 25 cities of 14 provinces,autonomous regions and municipalities in the whole country.Multi-stage stratified cluster sampling method was used.Again,using the same sampling method,4096 rural children (of whom 50.6％ were males with 6.2％ as minorities) were chosen from 26 cities of 10 provinces or municipalities.Identification of children being neglected was based on “Child Neglect Evaluation Norms of Children Aged 3-6 Years in Urban/Rural China”.SPSS-Windows 13.0 was employed for data analysis.Scores,frequency/degrees,age,sex and types (physical,emotional,educational,safety,medical and social) of children under negligence on every group of the regions,were calculated.x2 test (Chi-Square) and Analysis of variance (ANOVA) were processed to determine the significance of their differences.Results The overall frequencies of negligence were 28.0％ and 53.7％ respectively among the urban and rural children aged 3-6,while the total degrees of negligence were 42.2 and 44.4 respectively.Significant difference was found between children from the urban and the rural areas (P＜0.05).Significant difference was also found between urban and rural children on every age group (P＜0.05).The frequencies of negligence among males were 32.6％ and 55.9％ respectively in urban and rural areas while among females,the figures appeared to be 23.7％ and 51.6％ respectively.The degrees of negligence were 42.7 and 44.6 among male while 41.8 and 44.3 among female children,in the urban or rural areas.Significant differences were found on male or female between urban and rural groups (P＜0.05).Frequencies of negligence in urban children aged 3-6 for the six types were from 5.1％ to 12.9％,with the frequency in rural areas as 13.1％-26.6％.Significant difference was found between urban and rural group for any other type (P＜0.05),in addition to the safety type.The degrees of negligence in urban children aged 3-6 for the different type were between 39.4 and 43.4,while in the rural areas as from 36.5 to 48.2,with significant difference for every type (P＜0.05).The degrees of negligence related to education,emotion,or physical strength were more serious on children from the urban than from the rural areas.The highest frequency of child negligence was seen in the single-parent families on both urban and rural groups (42.9％ and 60.0％ respectively),with no significant difference found (P＞0.05).The urban and rural children aged 3-6 were mainly involved in single item of negligence,with incidence rates as 16.5％ and 22.7％ and proportions as 58.9％ and 45.1％ respectively,despite the factors as age or sex.Conclusion There were large differences on the situation of negligence between the urban and rural children aged 3-6.The frequencies and degrees of negligence in every age group and different sex for children living in the rural areas were higher than those urban children.The frequency of negligence among boys was higher than girls for both urban and rural areas.The rural children had suffered more serious negligence than the urban children at any other type,in addition to the ‘ safety'.Both urban and rural children had the highest frequency of negligence in single-parent family,and were mainly suffered from single item of negligence.

Analysis on the current situation of neglected rural children aged 0-6 vears and its impact factors in the western areas of China / 中华流行病学杂志

Objective To learn the current neglected situation and its impact factors on rural children in two provinces in Western China.Methods The investigation was conducted by using multistage stratified cluster sampling method in some parts of the Shaanxi province and the city of Chongqing.Results 1488 subjects,aged from 0 to 6,were recruited in the present study.Results showed that the total prevalence rates of neglected rural children in the two research sites were 31.59％ and 48.32 respectively.No significant difference was found on the prevalence of neglected for boys and girls ( x2 =0.86,U=1.51,P ＞ 0.05 ).The degrees of negligence in the older children showed a significant increase than in the younger children (x2 =13.36,F=33.45,P＜ 0.05 ).The prevalence and degree of negligence in families with three generations were lower than that of a single-parent families and remarried families (H=10.03,F=2.83,P＜0.05).The one-child-families were significantly lower than multiple children families,both in terms of prevalence and degree of negligence.Our data demonstrated the degrees of negligence in children whose parents were away from home,were higher than the children whose parents stayed with them ( x2=30.30,U=6.76,P＜0.05).The degree of negligence in Shaanxi was lower than that in Chongqing (x2=9.42,U=8.40,P＜0.05).Based on multivariate non-conditional logistic regression analysis,the determinants of stunting among the stranded children were as follows:parents were away from home (OR=1.54,95％ CI:1.20-1.97) ;structure of the family (OR=1.38,95％ CI:1.16-1.65) ; father' s occupation (OR=0.87,95％ CI:0.78-0.97) ; schooling of the children' s mother (OR=1.27,95％CI:1.07-1.52) ; relationship between children and their fathers (OR =1.43,95％ CI:1.07-1.91 ) etc.Conclusion The situation of negligence in children living in the rural areas,were serious in Shaanxi and Chongqing provinces,which called for the strengthening on the publicity and education of the issue.School and the society should also pay more attention to this problem.

Development of national neglect norms for children aged 3 to 6 years in rural areas of China / 中华预防医学杂志

<p><b>OBJECTIVE</b>To develop the national neglect norms for rural children aged 3 to 6 years, which are suitable for Chinese situations.</p><p><b>METHODS</b>According to the multi-stage stratified cluster sampling principle, 84 towns of 10 provinces or municipalities were selected in China. Children aged 3 to 6 years were surveyed in November 2010, the sample of analysis were 3240 (of whom males were 49.6% (1608/3240) and the Han nationality were 93.3% (3023/3240)). Questionnaire was designed by authors and deleted items that did not meet the requirements through several statistical analysis methods, such as item analysis method, factor analysis method, reliability analysis method. The reliability analysis and validity analysis were used to test the stability and reliability of the norm. The evaluation criteria of the scale was determined by the percentile method, then the initial development of the norm completed.</p><p><b>RESULTS</b>After deleting inappropriate items by statistical processing, finally, the scale consisted of 57 items, and included 6 neglected dimensions (physical neglect, emotional neglect, educational neglect, safe neglect, medical neglect and social neglect). Its item loadings ranged from 0.359 to 0.789, which met the statistical requirements. The scale's total Cronbach α coefficients 0.904, the total split-half reliability coefficients were 0.820, the 6 neglect dimensions' Cronbach α coefficients ranged from 0.620 to 0.815, the 6 neglect dimensions' split-half reliability coefficients ranged from -0.034 to 0.789, the scale's parallel reliability were 0.785 and it's re-test reliability were 0.613. After construct validity, external validity and content validity testing, the result showed that this scale could effectively reflect the real neglected status of children investigated. The total neglect cut-off score of this scale were 121.</p><p><b>CONCLUSION</b>The scale has good stability and reliability. And it adapts Chinese conditions and it's convenient to operate.</p>

The status of child neglect among rural areas children aged 0 - 6 years in China / 中华预防医学杂志

<p><b>OBJECTIVE</b>To explore the status of child neglect among rural areas children aged 0 - 6 years in China.</p><p><b>METHODS</b>A total of 7411 rural children aged 0 - 6 years old who were composed of two age groups (3315 children aged 0 - 2 years old and 4096 children aged 3 - 6 years old) were multistage stratified randomly sampled from 84 villages which were representative of 10 provinces of China, in accordance with sex and age in November 2010. To identify the child neglect based on the Neglect Norms for Children Aged 0 to 2 and 3 to 6 Years Old in Rural Areas of China, SPSS 13.0 was employed for analyzing neglect frequency and degree for every group of different age, sex and neglect type (including physical, emotional, educational, medical, safety and social neglects). χ(2) test and analysis of variance were also used.</p><p><b>RESULTS</b>The degree of child neglect for the children aged 0 - 2 years old was 45.01 ± 7.56, the neglect frequency was 54.9% (1819/3315); the degree of child neglect for the children aged 3 - 6 years old was 44.42 ± 7.57, the neglect frequency was 53.8% (2203/4096). The neglect frequency of children aged 0, 1, 2 years old were 58.5% (654/1117), 52.2% (597/1144), 53.9% (568/1054) (P < 0.05). For children aged 3 - 6 years old, the degrees of emotional and safety neglect for males (44.60 ± 7.86, 36.82 ± 9.03) were higher than females (44.03 ± 7.72, 36.25 ± 9.05) (P < 0.05); and the frequencies of emotional and social neglect for males (16.8% (349/2072), 28.3% (586/2072)) were also higher than females (14.1% (286/2024), 24.8% (503/2024)) (P < 0.05). All children of two age groups suffered neglect mainly on one of the six neglect types (incidences were 20.6% (683/3315) and 22.7% (931/4096)). For 0-2 age groups, the higher neglect frequencies happened in the single-parent family and the remarried family (62.5% (15/24) and 63.2% (12/19)), but for children aged 3 - 6 years old groups, it happened in the single-parent family (60.0%, 27/45).</p><p><b>CONCLUSION</b>Degree and frequency of child neglect among children aged 0 to 6 years old in the rural areas of China are high, and we should find out risk factors and provide efficient prevention measures.</p>

The status of child neglect for children aged 3 - 17 years in China / 中华预防医学杂志

<p><b>OBJECTIVE</b>To explore the status of child neglect among urban areas children and adolescents aged 3 - 17 years in China.</p><p><b>METHODS</b>Using multi-stage stratified cluster sampling method, 8001 children and adolescents were recruited from 30 cities of 14 provinces in December 2008. The investigation was carried based on The 3 - 6 years old children Neglect Norms of city in China and The 6 - 17 years old children Neglect Norms of city in China. SPSS 13.0 was employed for analyzing neglect rate and degree for group of areas, age, sex and neglect type (including neglect of physical, emotional, educational, medical, safety and social).</p><p><b>RESULTS</b>The degrees of child neglect for urban children aged 3 - 6 (children aged 6 were not in school), 6 - 8 (children aged 6 were school children), 9 - 11, 12 - 14 and 15 - 17 were 42.2 ± 6.9, 42.8 ± 8.4, 42.1 ± 9.1, 46.4 ± 9.2 and 49.7 ± 8.4(P < 0.05), the neglect rates were 28.0% (326/1163), 28.8% (431/1496), 27.2% (543/1962), 22.4% (373/1664) and 32.8% (563/1716) (P < 0.05), respectively. There were differences in six neglect types for each group; the neglect degrees and rates were high in the group aged 12 - 14 and 15 - 17 (46.4 ± 9.2 and 49.1 ± 8.4, 22.4% (373/1664) and 32.8% (563/1716), respectively); for each neglect type, the neglect degree of children aged 15 - 17 were highest (52.8 ± 8.0, 47.3 ± 11.6, 49.5 ± 10.8, 42.4 ± 10.3, 52.2 ± 16.0, 56.0 ± 10.1). The degree and rate of neglect in West were highest, Middle was next, the East was lowest (the neglect degrees were 45.4 ± 9.1, 45.0 ± 9.3 and 44.0 ± 8.8, P < 0.05;the rates were 30.7% (1340/4361), 25.0% (756/3024) and 21.2% (131/616), P < 0.05). Degree and rate of child neglect for children without siblings were lower than children with siblings (the neglect degrees were 44.2 ± 9.0 and 47.6 ± 9.1, P < 0.01; the rates were 26.1% (1572/6017) and 33.0% (655/1984), P < 0.01). Neglect degree of school children aged 6 years old (43.1 ± 7.3) was higher than children not in school (40.7 ± 7.8) (P < 0.01), but neglect rate was lower (the rates were 35.4% (60/169) and 21.3% (84/394), P < 0.01).</p><p><b>CONCLUSION</b>Neglect degree and rate of urban children and adolescents aged 3 - 17 years in China were both high, and neglect deserved more attentions.</p>

The current situation of children neglect and its influencing factors for rural children aged 0 - 6 years in Chongqing / 中华预防医学杂志

<p><b>OBJECTIVE</b>To learn the current situation of children neglect and its influencing factors among children aged from 0 - 6 years in rural Chongqing.</p><p><b>METHODS</b>Scales and evaluation methods in the "Chinese rural child neglected evaluation model" were used in this study. The investigation was conducted by using multistage stratified cluster sampling method in December 2010, and 822 rural children aged 0 to 6 years of three districts in Chongqing were randomly select for the survey, with neglect rate and the neglect degree to describe their neglect status, using logistic regression analysis to analyze factors affecting children neglect.</p><p><b>RESULTS</b>The total neglect rate and the neglect degree in the rural children were 35.64% (293/822) and 49.69 ± 6.45, respectively. The neglect degree in boys was higher than that of girls (boys: 50.23 ± 6.40 vs girls: 49.09 ± 6.47, P < 0.05). Further, Our data demonstrated both the neglect rate and the neglect degrees in children whose parents away from home were higher than that of the children whose parents stayed with them (neglect rate: 40.48% (202/499) vs 28.17% (91/323), P < 0.05; neglect degree: 50.64 ± 6.29 vs 48.23 ± 6.44, P < 0.05). Based on multivariate unconditional logistic regression analysis, the determinants of children neglect were: parents were away from home (OR = 1.61, 95%CI: 1.17 - 2.21), single mother and remarried family (OR = 1.39, 95%CI: 1.10 - 1.76), maternal occupation(OR = 0.81, 95%CI: 0.73 - 0.90), estrangement relationship between children and their mothers (OR = 1.42, 95%CI: 1.02 - 1.97).</p><p><b>CONCLUSION</b>The neglect situation of rural children is serious in Chongqing, and the main factors are parents away from home, single mother and remarried family, maternal occupation, and estrangement relationship between children and their mothers.</p>

The situation of children neglect between left-behind children and living-with-parents children in rural areas of two western provinces of China / 中华预防医学杂志

<p><b>OBJECTIVE</b>To understand the children neglect situation of left-behind children (children who do not grow up with their parents) and non-left-behind children in China's western rural and its influencing factors.</p><p><b>METHODS</b>Scales and evaluation methods in the "Chinese rural child neglected evaluation model" were used in this study. The investigation was conducted by using multistage stratified cluster sampling method; three countries were sampled randomly in Shanxi and Chongqing in November 2010, respectively. And, in every county, children from rural area aged from zero to six were randomly selected, with neglect rate and degree to describe their neglected status, using logistic regression analysis to analyze factors affecting the neglect rate.</p><p><b>RESULTS</b>Among 1568 subjects (859 left-behind and 709 living-with-parents), the total neglect rates were 29.78% (467/1568), and the total neglect degrees were 48.51 ± 6.49; the neglect rates for left-behind ones and living-with parents ones were 34.34% (295/859) and 24.26% (172/709) (P < 0.05); the neglect degree were 49.59 ± 6.54 and 47.19 ± 6.18 (P < 0.05). The neglect degree among left-behind ones and living-with parents ones between 0 to 2 years old were 48.59 ± 6.33 and 45.78 ± 5.94 (P < 0.05); in 3 to 6 years old group, which were 50.43 ± 6.60 and 48.25 ± 6.16(P < 0.05). The degrees in boy's group of these two kinds of children were 49.83 ± 6.67, 47.36 ± 6.28(P < 0.05) and girl's were 49.32 ± 6.38, 47.01 ± 6.08 (P < 0.05). On the other side, the neglect rate of left-behind and non-left-behind children between 0 to 2 years old were 39.33% (153/389) and 18.54% (56/302) (P < 0.05). The rates of boy's group were 34.91% (162/464) and 25.13% (94/374) (P < 0.05), and girl's were 33.67% (133/395) and 23.28% (78/335) (P < 0.05). Results showed that if the left-behind child's father was with lower education background, and the child and his mother feel stranger to each other, which led to the conclusion there was more chance for them to be neglected (OR values were 1.29 and 1.55, P < 0.05). If the non-neglected child's father was farmer or migrant worker and the relationship between the mother and father was poor, then there was more chance for them to be neglected (OR values were 0.85 and 1.92, P < 0.05).</p><p><b>CONCLUSION</b>The situation of children neglect in the western rural regions in China is serious. Both the neglect rate and degree among left-behind children are higher than those living-with-parents children. The influencing factors of neglect rate are different in the two groups.</p>

Construction, expression and identification of recombinant plasmid encoding bifunctional protein sflk1-IFN-gamma / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To construct, express and identify the recombinant plasmid pcDNA3.1(+)/sflk1-IFN-gamma encoding bifunctional protein sflk1-IFN-gamma (soluble fetal liver kinase 1 and interferon-gamma).</p><p><b>METHODS</b>sflk1 and IFN-gamma gene fragments were cloned by RT-PCR, and then inserted into pcDNA3.1(+) plasmid between BamHI-EcoRI and XhoI-XbaI restriction sites to form the recombinant plasmid pcDNA3.1(+)/sflk1-IFN-gamma. The recombinant sflk1-IFN-gamma transiently expressed in COS-7 cells was detected by ELISA and Western blotting. Bioactivities of sflk1-IFN-gamma fusion protein were identified by proliferation inhibition assay with H5V cells and NK activity assay.</p><p><b>RESULTS</b>pcDNA3.1(+)/sflk1-IFN-gamma can be effectively expressed in COS-7 cells. Concentrations of sflk1 and IFN-gamma in culture supernatants of pcDNA3.1(+)/sflk1-IFN-gamma transfected COS-7 cells were (20.85+/-2.48) ng/ml and (1.08+/-0.09) ng/ml, respectively. Western blotting showed that the molecular weight of sflk1-IFN-gamma fusion protein was about 130 kDa, while that of sflk1 was 115 kDa. The supernatants of transfected cells significantly inhibited the proliferation of H5V cells stimulated by mouse VEGF 164 and enhanced the NK activity of splenocytes, demonstrating that sflk1-IFN-gamma fusion protein possessed the bioactivities of both sflk1 and IFN-gamma.</p><p><b>CONCLUSION</b>The constructed plasmid pcDNA3.1(+)/sflk1-IFN-gamma can be effectively expressed in eukaryotes. The expressed sflk1-IFN-gamma fusion protein has the biological activities of both sflk1 and IFN-gamma.</p>

Research progress on expression and function of Toll-like receptors in dendritic cells / 浙江大学学报·医学版

Dendritic cells (DCs), the most potent antigen-presenting cells, recognize pathogen by Toll-like receptors (TLRs) and serve as the bridge between the innate and the adaptive immune responses. TLRs-mediated signal transduction plays a crucial role in the functional maturation of DCs. This review summarizes the research progress on the expression and function of TLRs in DCs.

Anti-metastatic effect of vascular endothelial growth factor receptor 2 extracellular domain gene-modified dendritic cell vaccination in murine model with experimental pulmonary metastasis / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the anti-metastatic effect of vascular endothelial growth factor receptor 2 extracellular domain gene-modified dendritic cell (DC-sVEGFR-2) vaccination.</p><p><b>METHODS</b>Dendritic cells (DC) were electroporated with pcDNA3. 1/sVEGFR-2 plasmid DNA. Expression of sVEGFR-2 was determined by ELISA. For immunization, C57BL/6 mice were intravenously injected three times with 1 x 10(5) cells per mouse of DC, pcDNA3. 1-transfected DC (DC-vector) , DC-sVEGFR-2, or 100 microl of PBS at 7-day intervals. At 10 days after the last immunization, the immunized mice were subjected to assessment of cytotoxic T lymphocyte ( CTL) response to VEGFR-2, alginate bead analysis of tumor cell-induced angiogenesis, and observation of the anti-metastatic effect in B16 melanoma metastasis model. CTL activity was determined by a standard 4-h 51Cr release assay against VEGFR-2 + vascular endothelial cell line H5V, 3LL cells stably transfected with pcDNA3. 1/sVEGFR-2 (3LL,-sVEGFR-2), and VEGFR-2- cell lines EL-4 and 3LL. Monoclonal antibodies GK1.5 anti-CD4 and 2.43 anti-CD8 were used to deplete in vivo CD4 + T cells and CD8' T cells, respectively.</p><p><b>RESULTS</b>DC-sVEGFR-2 could effectively express sVEGFR-2, whereas DC-vector and DC could not. Immunization of mice with DC-sVEGFR-2 significantly induce CTL activity against VEGFR-2 + cell lines H5V and 3LL-sVEGFR-2, however, no significant CTL activity was observed when VEGFR-2- syngeneic cell lines EL-4 and 3LL. were used as target cells, implying this CTL activity was VEGFR-2 specific. Alginate bead analysis of in vivo neoangiogenesis showed that the inhibition reached 50% in mice vaccinated with DC-sVEGFR-2 compared with mice vaccinated with DC, DC-vector or PBS. Anti-metastatic experiment showed that profound reduction in pulmonary metastases was found in mice immunized with DC-sVEGFR-2, while mice immunized with PBS, DC, DC-vector developed extensive pulmonary metastases. The number of tumor nodules on lung surface decreased by 81.9% in mice immunized with DC-sVEGFR-2 when compared with mice immunized with DC-vector (49.7+/-12.7 vs. 9.0+/-3.2). In vivo T cell subset depletion experiments showed that the anti-metastatic effect of DC-sVEGFR-2 vaccination was abrogated in CD8 + T cell-depleted but not in CD4+ T cell-depleted mice.</p><p><b>CONCLUSION</b>Immunization of mice with DC-sVEGFR-2 could break self-tolerance and induce a significant CTL response to VEGFR-2, leading to profound inhibition of tumor-cell induced angiogenesis and metastasis. This anti-metastatic effect is mainly mediated by CD8+ T cells.</p>

Study on the current situation and influential factors of child neglect among aged 3-6 year-olds in the urban areas of China / 中华流行病学杂志

<p><b>OBJECTIVE</b>Children aged 3 - 6 years in the urban areas of China were surveyed for the first time to find out the state of child neglect (CN) as well as the major relevant risk factors so as to provide evidence for developing intervention measures.</p><p><b>METHODS</b>1163 children (of whom 49.6% were males and 4.5% were minority nationality) were randomly sampled under multistage stratification, from 25 cities which representing 15 provinces of China. Based on the Child Neglect Norms used by China, prevalence of CN was identified and SPSS-Windows 11.0 was employed for statistical analysis. Scores, frequency/degrees, age, sex and 5 types (physical, emotional, educational, medical and safety) of CN on every group of the regions, were calculated. Multifactorial analysis was conducted through Binary Logistic Regression and multiple linear regression to determine the relevant risk factors.</p><p><b>RESULTS</b>(1) The average degree of CN for the 3 - 6 year-olds was 42.2, with its prevalence as 28.0%. Degrees of CN for the groups of 3, 4, 5, 6-year-olds were 41.7, 42.2, 42.1 and 43.1 (F = 0.988, P > 0.05), with frequencies of 25.0%, 25.3%, 27.9% and 35.4% (chi(2) = 4.798, P > 0.05), respectively. Degrees for CN in males and females were 42.7 and 41.8 (F = 2.502, P > 0.05) with the frequencies as 32.6% and 23.7% (chi(2) = 6.585, P < 0.05), respectively. Degrees of CN for the five types were 39.4-43.4 with the frequencies as 5.1%-12.9%, respectively. No significant difference was found in the frequency of the types (with an exception on 'physical neglect') between males and females (P > 0.05). The highest frequency (42.9%) of CN was seen in the single-parent families and the lowest in large family with three generations (25.5%). (2) According to monofactorial chi(2) test, the possible risk factors of CN would include: educational background, occupation and decrease of income of the parents during last year, etc. (3) Binary Logistic regression analysis showed that the influential factors to the occurrence of CN would include: father's educational background, sex of the child and mother's occupation, etc. (4) Multiple linear regression showed that the influential factors to the degree of CN were: family structure, number of supporting family members, relationship between parents and children, etc.</p><p><b>CONCLUSION</b>The degree and frequency of CN among children aged 3 to 6 in the urban areas of China were high but similar among the four age groups. Male children had a higher frequency of neglect than females, but with similar degree. Children in single-parent families had the highest frequency. The major influential factors of CN would include: educational background, occupation, family structure, family income of the parents which were similar to the results reported from foreign literature.</p>

Construction of recombinant plasmid containing mouse vasoactive intestinal peptide gene and its expression in COS-7 cells / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To construct the eukaryotic expression plasmid containing mouse vasoactive intestinal peptide(VIP) gene with biological activities.</p><p><b>METHODS</b>VIP cDNA including the sequences of signal peptide was cloned from mouse thymus by RT-PCR, and then inserted into the mammalian expression vector pcDNA3.1 between Hind III and EcoR I restriction sites. COS-7 cells were transfected with pcDNA3. 1-VIP using liposome, the expression of VIP was identified by Western blot and ELISA. Supernatant of transfected cell culture was added to LPS-stimulated macrophages and the TNF-alpha production in cell medium was observed by ELISA.</p><p><b>RESULTS</b>The cloned VIP cDNA was confirmed by enzyme digestion and DNA sequencing. The expression of VIP was detected in the pcDNA3. 1-VIP transfected COS-7 cells by Western blot and ELISA. The VIP in culture supernatant potently inhibited TNF-alpha production by LPS-induced Macrophages in vitro.</p><p><b>CONCLUSION</b>The eukaryotic expression plasmid that expresses biological active murine VIP has been constructed successfully.</p>

Investigation and analysis of child neglect in 1163 urban children aged 3 - 6 years / 中华儿科杂志

<p><b>OBJECTIVE</b>Child abuse and neglect (CNS) increasingly have become a serious problem in modern society all over the world. Children neglected and abused are at risk for maladaptation in behavioral, emotional, social, cognitive, and physical functioning. The purpose of this investigation was to develop the Child Neglect Scale for children aged 3 - 6 years in Chinese urban areas and to test the reliability and validity of this norm.</p><p><b>METHODS</b>According to the principle and procedure of developing scales, a child neglect questionnaire was designed. Multistage sampling was used in the study in accordance with sex, age, the levels of economy and kindergartens, a random sample of 1,465 children aged 2 - 6 years was selected from 25 cities. Of the 1,465 subjects who completed the Child Neglect Questionnaires again 2 weeks later, 66 had completed data on each of the two occasions. The Child Neglect Questionnaires were completed by parents and caregivers. The data were analyzed using items analysis, factor analysis, internal consistency analysis and the reliability and validity of the norm were tested. The factor structure of the Child Neglect Scale was determined using an oblique rotation. The internal consistency was reflected in Cronbach's alpha. Since no existing categorization scheme on child neglect could be found in the literature, one was formulated in an attempt to efficiently and meaningfully group similar types of caregiver behaviors. The initial pool of potentially neglecting caregiver behaviors was drawn from literature. The most common patterns of parental behaviors were grouped according to six neglect subscale schemes found in the literature, with modifications made to increase item specificity and reduce overlap. The initial Child Neglect Questionnaire contained a 25-item physical neglect subscale, a 59-item emotional neglect subscale, a 23-item educational neglect subscale, a 20-item safety neglect subscale, a 10-item medical neglect subscale, a 9-item social neglect subscale.</p><p><b>RESULTS</b>Completed Child Neglect Questionnaires were obtained from 1,457 subjects, 726 males and 731 females. The overall findings of this study came from 1,163 subjects, because children aged 2 years (294 subjects) were removed from subjects by statistical method. The 91-item Child Neglect Scale was presented to the respondent as a child neglect questionnaire with a 9-item social neglect subscale and some items in other subscales were removed from the initial questionnaire by using statistical method. The questionnaire contained 17 physical neglect items, 40 emotional neglect items, 17 educational neglect items, a 10 safety neglect items, and 7 medical neglect items. The internal consistency of the overall Child Neglect Scale as reflected in Cronbach's alpha was 0.94, the split-half reliability was 0.88, the test-retest reliability was 0.92. For physical, emotional, educational, safety, medical neglect subscale, Cronbach's alpha, respectively was 0.80, 0.88, 0.84, 0.68, 0.60, the split-half reliability was 0.54 - 0.88, the test reliability was 0.89, 0.94, 0.87, 0.80, 0.77, respectively. The construct validity, face validity and exterior validity of the norm were good. For physical, emotional, educational, safety, medical neglect subscale and overall child neglect scale, the scores of percentiles 90 were used as cut-point. They were 35, 80, 40, 20, 15, 190, respectively.</p><p><b>CONCLUSIONS</b>The norm was consistent with China's economic-cultural background and reality. It not only may be used for research purpose but also may be useful in assessing parent's and child-oriented individual's behaviors and attitude as a measuring instrument.</p>