FOXO4 maintains senescence in human umbilical cord mesenchymal stem cells by repressing apoptosis / 生理学报

The purpose of the present study was to investigate the effects of forkhead box O4 (FOXO4) on the senescence of human umbilical cord-derived mesenchymal stem cells (hUC-MSCs). The hUC-MSCs were induced to senescence by natural passage, and FOXO4 expression was inhibited by lentiviral shRNA transfection. The hallmark of cell senescence was analyzed by β-galactosidase staining, and the cell viability was assayed by CCK-8 method. Flow cytometry was used to investigate the apoptosis of hUC-MSCs. The expression levels of Bcl-2, Bax, FOXO4, interleukin 6 (IL-6) and cleaved Caspase-3 were detected by qPCR and Western blot. Immunofluorescence staining was used to detect FOXO4 expression. The amount of IL-6 secreted by hUC-MSCs was detected by ELISA. The results showed that, compared with the passage 1, senescent hUC-MSCs showed up-regulated expression levels of Bax and FOXO4, down-regulated expression levels of Bcl-2 and cleaved Caspase-3, and increased IL-6 mRNA expression and secretion. FOXO4 inhibition in senescent hUC-MSCs promoted cell apoptosis, reduced cell viability, and inhibited the mRNA expression and secretion of IL-6. These results suggest that FOXO4 maintains viability and function of senescent hUC-MSCs by repressing their apoptosis response, thus accelerating senescence of the whole cell colony.

Current status of neglect among children aged 3-6 years in rural areas of Urumqi, China and risk factors for child neglect / 中国当代儿科杂志

OBJECTIVE@#To investigate the current status of neglect in children aged 3-6 years in the rural areas of Urumqi, China and risk factors for neglect.@*METHODS@#Stratified random cluster sampling was performed to select 1 326 children, aged 3-6 years, from 6 kindergartens in the rural areas of Urumqi. A self-designed questionnaire and "Chinese Neglect Evaluation Questionnaire for Rural Children Aged 3-6 Years" were used for investigation.@*RESULTS@#Of the 1 326 children, the neglect rate was 51.89% (688 cases), and the degree of neglect was 49±6. There were significant differences in the neglect rate and the degree of neglect between the children in different age groups (P<0.01). The single-parent families and the remarried families had significantly higher child neglect rate and degree of neglect than the families of three generations under one roof and the core families (P<0.01). The children in left-behind status had a significantly higher degree of neglect (P<0.01). The multivariate logistic regression analysis showed that younger children, mothers with low education level, employment status of the mother as a migrant worker, single-parent and remarried families, low-income families, and left-behind status were risk factors for child neglect (P<0.05).@*CONCLUSIONS@#The neglect of children aged 3-6 years in the rural areas of Urumqi is serious, and more attention should be paid to younger children, children whose mothers with low education level, left-behind children, and children from single-parent families, remarried families and low-income families.

Current status of social anxiety among primary school students with myopia in Urumqi, China and risk factors for myopia / 中国当代儿科杂志

OBJECTIVE@#To investigate the prevalence of social anxiety among the fourth-, fifth- and sixth-grade primary school students with myopia in Urumqi, China and the risk factors for social anxiety.@*METHODS@#Stratified cluster random sampling was used to select 552 fourth-, fifth- and sixth-grade primary school students with myopia from four primary schools in Urumqi. A self-designed questionnaire, a social anxiety scale for children and a self-esteem scale were used to investigate the general demographic data and the current status of social anxiety and self-esteem. A multivariate logistic regression analysis was used to investigate the risk factors for social anxiety in primary school students with myopia.@*RESULTS@#Of the 552 children, 173 (31.3%) were found to have social anxiety. The multivariate logistic regression analysis showed that a higher grade, female sex, autocratic family, high myopia, low self-esteem and wearing glasses for more than 2 years were risk factors for social anxiety (P<0.05).@*CONCLUSIONS@#There is a serious problem of social anxiety among the fourth-, fifth- and sixth-grade primary school students with myopia in Urumqi. The development of social anxiety is associated with age, sex, degree of myopia, time of wearing glasses, parental education style and self-esteem level.

Molecular mechanism of polygonatum sibiricum polysaccharide in the prevention and treatment of postmenopausal osteoporosis / 中国组织工程研究

BACKGROUND: Previous studies have found that polygonatum sibiricum polysaccharide (PSP) exhibits anti-osteoporosis effect, but its therapeutic effect in ovariectomized osteoporotic rats and the molecular mechanisms are poorly understood. OBJECTIVE: To investigate the effect of administration of PSP on the bone microstructure, bone mineral density as well as osteoblast- and osteoclast-related gene expression in rats. METHODS: Twenty-five infertile female Sprague-Dawley rats aged 3 months were randomly allotted into five groups (n=5 per group): sham operation (same volume normal saline), model, zoledronate (0.2 mg/kg?d), high-dose PSP (800 mg/kg?d) and medium-dose PSP (400 mg/kg?d) groups. All rats were subjected to ovariectomy except sham operation group. The administration was intragastrically given every 2 days beginning at 7 days after modeling and lasted 12 weeks. Then, the rats were sacrificed, and the uterus was weighed. The bilateral tibias were removed, one side for histomorphometric analysis by micro-CT, and the other one for RNA detection by qualified PCR. RESULTS AND CONCLUSION: Compared with the sham operation group, the rat body mass in the model group was significantly increased and the weight of uterus was significantly decreased (P < 0.05). Compared with the model group, zoledronate and high-dose PSP could significantly alleviate the excessive increase in body mass (P < 0.05). The bone mineral density in the model group was decreased by 63% compared with the sham operation group (P < 0.01), Compared with the model group, after 12-week high-dose PSP and zoledronate administration, the bone mineral density was increased by 44% and 38%, respectively (P < 0.01); the trabecular bone volume fraction and trabecular number rose significantly(P<0.05),while the trabecular separation decreased significantly(P<0.05).In vivo,PSP could significantly promote the expression levels of osteoblast-related genes (alkaline phosphatase, RUNX2, Col1a1 and osteocalcin), and significantly inhibit the expression levels of osteoblast-related genes (ACP5 and CTSK) (P < 0.05). These results imply that high-dose PSP can reduce bone loss and decrease of bone mineral density, improve the destruction of bone microstructure, as well as promote osteoblast-related genes but inhibit osteoclast-related gene mRNA expression in the ovariectomized rats.

Current status of social anxiety and depression among primary school students in Urumqi, China / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the current status of social anxiety and depression among the fourth-, fifth-, and sixth-grade students in Urumqi, China, and to provide data support for mental health education for primary school students in the future.</p><p><b>METHODS</b>Stratified cluster random sampling was performed to select 919 fourth-, fifth-, and sixth-grade students from four primary schools for the Han and minority ethnic groups in Urumqi as research subjects. Social Anxiety Scale for Children and Children's Depression Inventory were used to evaluate the current status of social anxiety and depression.</p><p><b>RESULTS</b>Among the 919 fourth-, fifth-, and sixth-grade students, the detection rate of social anxiety was 28.5% (262 students). The fourth-grade students had significantly lower scores on each subscale of social anxiety and total score of social anxiety than the fifth- and sixth-grade students (P<0.05). Girls had significantly higher score on the subscale of fear of negative evaluation and total score of social anxiety than boys (P<0.05). The Uyghur students had significantly higher scores on each subscale of social anxiety and total score of social anxiety than the Han students and the students of other minority ethnic groups (P<0.05). Among these fourth-, fifth-, and sixth-grade students, the detection rate of depression was 11.2% (103 students). The fourth-grade students had significantly lower scores on the subscales of negative self-esteem, ineffectiveness, and interpersonal problems and total score of depression than the fifth- and sixth-grade students (P<0.05). The Uyghur students had significantly higher scores on the subscales of anhedonia and negative mood and total score of depression than the Han students and the students of other minority ethnic groups (P<0.05).</p><p><b>CONCLUSIONS</b>There are significant differences in the status of social anxiety and depression among the primary school students with different grades, sexes or ethnic groups in Urumqi. The fifth- and sixth-grade students, girls, and Uyghur students tend to have more serious psychological problems, which should be taken seriously by parents, schools, and the society.</p>

Current status of parental feeding behaviors in Urumqi, China, and its association with body mass index of children / 中国当代儿科杂志

OBJECTIVE@#To investigate the association between parental feeding behaviors and body mass index (BMI) of children, and to provide a scientific basis for the prevention and intervention of overweight and obesity in children.@*METHODS@#Stratified random cluster sampling was used to select 976 children from 7 kindergartens in Xinshi District of Urumqi, and a questionnaire survey and physical measurement were performed to obtain related data.@*RESULTS@#A total of 976 questionnaires were distributed and 924 (94.7%) usable questionnaires were collected. The overall detection rates of underweight, overweight, and obesity in children were 3.1% (29 children), 9.2% (85 children), and 6.7% (62 children), respectively. The highest level of parental feeding behavior was monitoring, followed by restriction and pressure to eat. The parents of the Han children had significantly higher levels of restriction and pressure to eat than those of the Uygur children (P<0.01). As for parental feeding behaviors, restriction to eat was positively correlated with BMI of the Han and Uygur boys (P<0.01); pressure to eat was negatively correlated with BMI of the Han boys and girls (P<0.01) and was positively correlated with BMI of the Uygur boys and girls (P<0.01); monitoring was negatively correlated with BMI of the Han and Uygur boys and girls (P<0.05). Among the parents of the Han and Uygur boys, the parents of the overweight/obese children had significantly higher scores of restriction to eat than those of the children with normal weight (P<0.05); among the parents of the Uygur boys and girls, the parents of the overweight/obese children had significantly higher scores of pressure to eat than those of the children with normal weight (P<0.01); among the parents of the Uygur and Han boys and girls, the parents of the overweight/obese children had significantly lower scores of monitoring than those of the children with normal weight (P<0.01).@*CONCLUSIONS@#The parental feeding behavior is good in Urumqi, and the parental feeding behavior of the Uygur children is a little better than that of the Han children. Parental feeding behavior is closely correlated with BMI of children, and such correlation differs across ethnic groups and sexes. A high level of monitoring and low levels of restriction and pressure to eat may help to prevent and control the development of overweight and obesity in children.

Regulation of SIRT1 by SREBP-1 in Adipogenic Differentiation of Mesenchymal Stem Cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the regulation of SIRT1 by transcription factor SREBP-1 in adipogeneic differentiation of bone marrow mesenchymal stem cells (BMMSC).</p><p><b>METHODS</b>Oil red O staining was used to identify the adipogenic differentiation of BMMSC; the mRNA transcription levels of AP2, LPL, SREBF-1, SIRT1 gene were detected by RT-PCR; the expession level of SREBP-1 was determined by Western-blot. The chromatin immunoprecipitation (ChIP) assay was used to investigate the binding of SREBP-1 to SIRT1 promoter.</p><p><b>RESULTS</b>BMMSC exposed to adipogenesis inducing medium become mature adipocytes at day 14; the mRNA transcription levels of AP2, LPL, SREBF-1, SIRT1 genes were up-regulated in adipocyte differentiation of BMMSC; the protein level of SREBP-1 was higher obviously; SIRT1 gene sequences was succesfully amplified from the genomic DNA immunoprecipitated by SREBP-1 antibody.</p><p><b>CONCLUSION</b>SREBP-1 can bind to the promoter region of the SIRT1 gene in adipogenesis of BMMSC, and may be involved in the transcriptional regulation of the SIRT1 gene.</p>

Effects of Leukemia Inhibitory Factor Combined with Basic Fibroblast Growth Factor on Self-maintenance and Self-renewal of Human Umbilical Cord Mesenchymal Stem Cells In Vitro / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the effects of LIF combined with bFGF on the proliferation, stemness and senescence of hUC-MSC.</p><p><b>METHODS</b>Experiments were divided into 4 groups: control group, in which the cells were treated with complete medium (α-MEM containing 10% FBS); group LIF, in which the cells were treated with complete medium containing 10 ng/ml LIF; group bFGF, in which the cells were treated with complete medium containing 10 ng/ml bFGF; combination group, in which the cells were treated with complete medium containing 10 ng/ml LIF and 10 ng/ml bFGF. The growth curves of hUC-MSC at passage 4 in different groups were assayed by cell counting kit 8. Cellular morphologic changes were observed under inverted phase contrast microscope; hUC-MSC senescence in different groups was detected by β-galactosidase staining. The expression of PCNA, P16, P21, P53, OCT4 and NANOG genes was detected by RT-PCR.</p><p><b>RESULTS</b>The cell growth curves of each group were similar to the S-shape; the cell proliferation rate from high to low as follows: that in the combination group > group bFGF > group LIF > control group. Senescence and declining of proliferation were observed at hUC-MSC very early in control group; the cells in group LIF maintained good cellular morphology at early stage, but cell proliferation was slow and late senescence was observed; a few cells in group bFGF presented signs of senescence, but with quick proliferation; the cells in combination group grew quickly and maintained cellular morphology of hUC-MSC for long time. The LIF and bFGF up-regulated the expression of PCNA, OCT4 and NANOG, while they down-regulated the expression of P16, P21, P53, and their combinative effects were more significant.</p><p><b>CONCLUSION</b>LIF combined with bFGF not only can promote the proliferation and maintenance of stemness of hUC-MSC, but also can delay the senescence of hUC-MSC.</p>

Fingerprint analysis of Anoectochili Roxburghii Gemma Terminalis by HPLC / 中草药

Objective: To establish the chromatographic fingerprint analysis for the quality of Anoectochili Roxburghii Gemma Terminalis. Methods: The separation was performed on an Agilent TC-C18 column (250 mm × 4.6 mm, 5 μm) with a gradient elution composed of methanol and water. The detective wavelength was at 260 nm. The flow rate was 1.0 mL/min and the column temperature was set at 30℃. Results: Twenty-two characteristic peaks were established in the fingerprint. The mutual model of Anoectochili Roxburghii Gemma Terminalis was established and the similarities were calculated. Seven chromatographic peaks among twenty-two common peaks were identified by LC-MS/MS. Ten batches of Anoectochili Roxburghii Gemma Terminalis were determined and the similarities were above 0.9. Conclusion: The method is accurate with good reproducibility. It can be used for the quality control of Anoectochili Roxburghii Gemma Terminalis.

Expression and significance of EZH2 and DNMT1 in triple-negative breast cancer / 解放军医学杂志

Objective To probe into the expression features of EZH2 and DNMT1 in triple-negative breast cancer (TNBC), and the relations between the expression and the main clinical pathological parameters of TNBC. Methods The clinical and pathological data of 209 cases of breast cancer (including 131 cases of TNBC and 78 cases of non-TNBC) in Guangzhou Kingmed Center for Clinical Laboratory from Jan. 2008 to May 2013 were retrospectively analyzed. Immunohistochemistry SP assay was performed to determine the expressions of EZH2 and DNMT1 in 209 paraffin-embedded specimens of breast cancer and 65 specimens of normal tissue (more than 4cm away from tumor). The differences of protein expression in the specimens were compared, and correlation analysis was performed to disclose the relationship between protein expression and clinico-pathological indices. Multivariate logistic regression analysis was applied to analyze the positive expression-related factor of EZH2 and DNMT1. Spearman rank correlation analysis was used to analyse the interrelation of EZH2 and DNMT1 in TNBC. Results The positive expression rates of EZH2 in TNBC, non-TNBC and adjacent breast tissue were 86.3%, 89.7% and 40.0%, respectively, while the positive expression rates of DNMT1 were 63.4%, 66.6% and 44.6%, respectively. The positive expression rates of EZH2 and DNMT1 were higher in both TNBC and non-TNBC tissues than in adjacent breast tissue (P0.05), but the positive expression of EZH2 in TNBC was related to the pathological grade, tumor size and lymph node metastasis (P<0.05), and the positive expression of DNMT1 was related to the pathological grade and lymph node metastasis (P<0.05). Multivariate logistic regression analysis demonstrated that the pathological grade of tumor was the main factor affecting the expressions of EZH2 and DNMT1, and the expressions of EZH2 and DNMT1 in TNBC were positively correlated (r=0.34, P<0.01). Conclusion The high expressions of EZH2 and DNMT1 are correlated with the differentiation, growth and metastasis of TNBC. Combined detection of the expressions of EZH2 and DNMT1 may be more valuable in determination of prognosis of patients with TNBC.

Tissue culture regeneration and genetic transformation of Coptis chinensis / 中草药

Objective: To establish the system of tissue culture regeneration and gene transformation for Coptis chinensis. Methods: The cotyledon and hypocotyl of C. chinensis were used as explants, and the effects of different basic media with different plant growth regulators on in vitro tissue culture regeneration were compared; The embryogenic calli of C. chinensis were used as recipients for genetic transformation by particle bombardment method. Results: The induction rates of cotyledon and hypocotyl were 86.31% and 54.34%, respectively; The 6, 7-V medium was more suitable for callus induction and proliferation rate of C. chinensis than the MS medium. The optimal hormone combination for callus induction was 0.5 mg/L 2, 4-D + 0.5 mg/L KT in MS medium; Either the appropriate concentration of cytokinin alone or the combination of both cytokinin and auxin could induce the continuous proliferation of calli; However, only the hormone combination 0.5 mg/L KT + 0.5 mg/L IAA and 1.0 mg/L 6-BA + 0.5 mg/L NAA could generate the somatic embryos. 6, 7-V medium in absence of hormone could maintain the continuous growth of the embryogenic calli. The somatic embryos could germinate and grow up to plantlets on 6, 7-V medium containing 1 mg/L GA3 + 0.5 mg/L IBA. The embryogenic calli were transformed by particle bombardment method and screened under 3 mg/L Basta, then the activity of β-glucuronidase was detected. PCR analysis showed that the bar gene was successfully transferred to the regenerated plants. Conclusion: The tissue culture regeneration and genetic transformation system of C. chinensis is established, which lays the foundation for its genetic improvement.

Effects of nandrolone phenylpropionate on ultrastructure of aorta in rats with or without movement training / 上海第二医科大学学报

Objective To observe the influence of nandrolone phenylpropionate(NP) on the ultrastructure of aorta in rats with or without movement training,and investigate the side effects of NP on the cardiovascular system. Methods Forty male SD rats were randomly divided into sedentary control group,sedentary+medicine group,exercise control group and exercise+medicine group.For the groups with medical treatment,NP of 10 mg/kg one time every three days was injected into the rats via gluteus for eight weeks.For the exercise groups,rats were trained to run on treadmill five days per week for eight weeks.The aortae were sampled and specimens were obtained for transmission electron microscopy. Results The ultrastructure of aorta was normal in sedentary control group.For sedentary+medicine group,mitochondrial swelling,vacuolated cytoplasm and lysis of endothelial cells were observed,disruption of intercellular conjunctions,widening of subendothelial spaces and furcation and breakage of internal elastic lamina were found,and smooth muscle cells changed into synthesis type.For exercise control group,no obvious morphologic change was observed,except that part of the internal elastic lamina disrupted.In exercise+medicine group,breakage and lysis of endothelial cells were observed,widening of subendothelial spaces and lysis of internal elastic lamina were found,and autophagosome and myelinoid body were seen in smooth muscle cells. Conclusion NP may lead to the impairment of endothelial cells and the change of smooth muscle cells into synthesis type.Exercise with NP administration may result in more severe impairment in vessel wall.

Expression of CDK4 and cyclin D1 proteins in gastric carcinoma / 上海第二医科大学学报

Objective To investigate the expression and significance of CDK4 and cyclin D1 in gastric carcinoma,and to discuss their correlation with WHO histological classification,TNM stages and lymph node metastasis.Methods Forty paraffin wax specimens from patients with gastric carcinoma were collected and expression of CDK4 and cyclin D1 proteins were detected by immunohistochemistry.Results Immunohistochemistry demonstrated that the positive rates for CDK4 and cyclin D1 protein were 65% and 55%,respectively.There was positive correlation between CDK4 and cyclin D1 proteins(P0.05).Conclusion CDK4 and cyclin D1 proteins are frequently overexpressed in gastric carcinoma and there is a positive relationship between the two proteins,which may be involved in the development of gastric carcinoma.