The effect of China's birth policy changes on birth defects-A large hospital-based cross-sectional study.

A retrospective analysis of birth data hospital-based obtained from 14 monitoring areas in the Huaihe River Basin from 2009 to 2019 was conducted. Trend in the total prevalence of birth defects (BDs) and subgroups were analyzed using the Joinpoint Regression model. The incidence of BDs increased gradually from 118.87 per 10,000 in 2009 to 241.18 per 10,000 in 2019 (AAPC = 5.91, P < 0.001). Congenital heart diseases were the most common subtype of BDs. The proportion of maternal age younger than 25 decreased but the age 25-40 years increased significantly (AAPC<20=-5.58; AAPC20-24=-6.38; AAPC25-29 = 5.15; AAPC30-35 = 7.07; AAPC35-40 = 8.27; All P < 0.05). Compared with the one-child policy period, the risk of BDs was greater for groups among maternal age younger than 40 years during the partial and universal two-child policy period (P < 0.001). The incidence of BDs and the proportion of women with advanced maternal age in Huaihe River Basin is increasing. There was an interaction between changes in birth policy and the mother's age on the risk of BDs.

DietaryAdvancedGlycationEnd Products-InducedCognitive Impairment in Aged ICR Mice: Protective Role of Quercetin.

SCOPE: Dietary advanced glycation products (dAGEs) have been reported to induce cognitive impairment while quercetin possesses potential neuroprotective effects. The aim is to explore whether dAGEs would induce similar cognitive impairment from both young and aged ICR mice, and the protective effects of quercetin. METHODS AND RESULTS: A total of 32 aged ICR mice (15-month-old) and 16 young ICR mice (3-month-old) are randomly assigned into the following six groups: Young mice control group, young mice fed with AGEs diet group, old mice control group, old mice fed with AGEs diet group, old mice with quercetin supplemented diet group, old mice fed with AGE diet supplemented with quercetin group. Dietary AGEs induced cognitive impairment only in aged, but not in young, ICR mice, while quercetin intervention is capable of reversing dAGEs-induced cognitive dysfunction. This may be since quercetin 1) increased miR-219, miR-15a, and miR-132 expression, inhibited p-ERK1/2, and tau phosphorylation; and 2) improved gut microbiota richness and diversity, inhibited phylum Tenericutes and Proteobacteria, and elevated butyric acid from cecum. CONCLUSION: Prolonged application of quercetin may be beneficial in the elderly, especially for those with high consumption of dAGEs.

Grape seed proanthocyanidin extract alleviates high-fat diet induced testicular toxicity in rats.

The present study aimed to investigate the protective effects of grape seed proanthocyanidin extract (GSPE) on high-fat diet (HFD) induced testicular damage, oxidative stress, and apoptotic germ cell death. Male rats (n = 40) were randomly divided into four groups: the control group (treated with physiological saline), HFD group, HFD + GSPE (100 mg kg-1) group and HFD + GSPE (300 mg kg-1) group. Compared with the HFD group the rats of the GSPE-treated group showed improved serum testosterone levels, sperm quality and histological appearance of the testis tissue. Significant elevation of antioxidant enzyme (SOD, GSH, and GSH-Px) activities and remarkable reduction in MDA were also observed by GSPE administration, indicating that GSPE can decrease testicular oxidative stress. Finally, a significant reduction in spermatogenic cell apoptosis was detected by TUNEL assay. In summary, these results indicated that GSPE can suppress testicular dysfunction and this effect may be attributed to its antioxidant and anti-apoptotic properties. The current study indicates that GSPE can be considered a promising candidate for use as a drug or a food supplement to alleviate HFD-induced testicular dysfunction.

Maternal Lead Exposure Induces Down-regulation of Hippocampal Insulin-degrading Enzyme and Nerve Growth Factor Expression in Mouse Pups.

Lead exposure is a known potential risk factor for neurodegenerative diseases such as Alzheimer's disease (AD). Exposure to lead during the critical phase of brain development has been linked with mental retardation and hypophrenia in later life. This study was aimed to investigate the effects of lead exposure of pregnant mice on the expressions of insulin-degrading enzyme (IDE) and nerve growth factor (NGF) in the hippocampus of their offspring. Blood samples were collected from the tail vein, and after anesthetizing the pups, the brain was excised on postnatal day 21. Lead concentrations were determined by graphite furnace atomic absorption spectrophotometry, and the expressions of IDE and NGF were determined by immunohistochemistry and Western blotting. Results showed that the reduction in IDE and NGF expression in the hippocampus of pups might be associated with impairment of learning and memory and dementia induced by maternal lead exposure during pregnancy and lactation.

[Effect of pueraria crude extreact and puerarin on ethanol-induced expression of heat shock protein 70 in embryonic mouse hippocampal cultures].

OBJECTIVE: To study if the Pueraria crude extreact (CP) and standard preparation of pure puerarin (SP) possess the same neuroprotective effects on the expression of heat shock protein (HSP) 70 in the embryonic mouse hippocampal cells. METHODS: The hippocampus of 18-days-old mouse embryo was taken out and suspension of single cells was cultured. Ethanol was added to cause HSP70 mRNA expression. Solvent, ethanol of different concentrations (50, 200, and 300 mmol/L), SP + ethanol, and SP + ethanol were added respectively. Western blotting was used to detect the expression of the expression of HSP70 mRNA. RESULTS: Ethanol of different concentrations increased the expression of HSP70 mRNA and the protein in comparison with the solvent control group. SP and CP inhibited the expression of HSP70 mRNA and protein. CONCLUSION: With identical effect of anti-oxidative stress, both SP and CP inhibit the increase of expression of HSP70 mRNA and protein, thus demonstrating I vitro anti-oxidative neuroprotection.

[Retinoic acid induced cell cycle arrest and apoptosis in mouse embryonic palatal mesenchymal cells].

OBJECTIVES: To investigate the effect of all-trans retinoic acid (atRA) on proliferation activity and cell cycle distribution in mouse embryonic palatal mesenchymal (MEPM) cells and the underlying molecular mechanisms. METHODS: MEPM cells were prepared from palate shelves of mouse fetal on gestation day 13. Cell viability was determined by MTI assay. Cell cycle distribution and subdiploid population were analyzed by cytometry. The expression of cyclin D and E and phosphorylation of retinoblastoma protein was examined using Western-blot. RESULTS: atRA remarkably inhibited the growth of MEPM cells in a dose-dependent manner. atRA also caused an increase in the proportion of cells in G0/G1 and a decrease in the proportion of cells in S phase. atRA inhibited expression of cyclins D and E at protein level. Furthermore, atRA treatment reduced phosphorylated Rb. CONCLUSION: These data suggested that atRA had antiproliferative activity by modulating G1/S cell cycle regulators and by inhibition of Rb phosphorylation in MEPM cells, which might account for the pathogenesis of cleft palate induced by retinoic acid.

[Effects of genistein on proliferation and apoptosis in HT-29 cells].

OBJECTIVE: The study is to explore the effects of genistein on proliferation and apoptosis in human colon cancer HT-29 cells and the likely underlying molecular mechanisms. METHODS: HT-29 cultures were maintained in DMEM containing 10% fetal bovine serum. Cell proliferation was determined by MTT assay and cell cycle distribution by cytometry. Apoptosis was detected by the Cell Death Detection ELISA and cytometry. The expressions of bax, bcl-2, and PCNA were examined using reverse transcriptase-polymerase chain reaction (RT-PCR) and Western-blot both at mRNA and protein levels, respectively. RESULTS: Genistein inhibited proliferation and induced G2/M phase arrest and apoptotic death in colon cancer HT-29 cells. We investigated the effects of genistein on molecules that regulate apoptosis and cell cycle progress. Genistein increased expression of bax and significantly reduced PCNA with a slightly decrease in bcl-2 expression both at mRNA and protein level. CONCLUSION: Our results demonstrated that genistein inhibited the viability of human colon cancer HT-29 cell via induction of apoptosis mainly through regulation of PCNA and Bax/Bcl-2 expression. These data suggested a role of genistein in prevention of colon tumor and might reduce colon tumor growth.

[Potential antiviral drug Pueraria crude extract and puerarin protect against ethanol-induced cytotoxicity in embryonic mouse hippocampal cultures].

OBJECTIVE: To examine whether Chinese medical herb Pueraria crude extract (CP) and standard of pure puerarin (SP) possess the same neuroprotective effects during concomitant ethanol (EtOH) treatment. METHODS: Hippocampus cultures were prepared from mice at gestational age of 18 day. Cell viability was measured by MTT assay. RT-PCR was employed to determine mRNA expression of superoxide dismutase (SOD). RESULTS: As measured by MTT assay, supplementation with 15 mg/L CP or 10 mg/L SP afforded neuroprotection against all EtOH concentrations (50, 200 and 350 mmol/L, respectively) in embryonic hippocampal culture system. In addition, both 15 mg/L CP and 10 mg/L SP could decrease expression of SOD at mRNA level. CONCLUSION: This study suggests that CP and SP could decrease oxidative stress induced by ethanol treatment by the decreased expression of SOD at mRNA level, and demonstrates antioxidative neuroprotective effect of CP and SP against developmental ethanol exposure in vitro.

[Effects of zearalenone on proliferation and apoptosis in MCF-7 cells].

OBJECTIVE: To explore the effects of zearalenone (ZEA) on proliferation and apoptosis in estrogen-dependent human breast cancer MCF-7 cells and the likely underlying molecular mechanisms. METHODS: Cell viability was determined by MTT assay and cell cycle distribution by cytometry. Apoptosis was detected by Cell Death Detection ELISA and cytometry, respectively. The expressions of bax and bcl-2 were examined using multiple RT-PCR and Western-blot both at mRNA and protein level, respectively. RESULTS: The current study confirmed the previous studies that ZEA could stimulate proliferation in MCF-7 cells with inducing a profound increase in S phase and a modest increase in G(2)/M phase that was accompanied by a decrease in G(0)/G(1) phase. ZEA could inhibit apoptosis in MCF-7 cells following estrogen ablation at a range of concentrations of 2 nmol/L -96 nmol/L. Western blot and RT-PCR analysis revealed that the anti-apoptotic bcl-2 was upregulated at both protein and mRNA level, together with the downregulation of pro-apoptotic bax. CONCLUSION: ZEA should have possessed comparative estrogenic activity and could promote the progression of MCF-7 cells through the cell cycle by a decreasing in the G(0)/G(1) phase and by a significant increasing in S-phase. The pro-proliferative activity of ZEA was due to inhibition of apoptosis through regulation of bax/bcl-2 expression.

[The effects of three plastic additives on the proliferation of MCF-7 cell].

OBJECTIVE: To explore the effect of environmental estrogens on the proliferation of breast cancer cell line MCF-7. METHODS: The tested compounds were n-4-nonyphenol, Bisphenol A and dibutylphthalate. Human estradiol-dependent MCF-7 breast cancer cells were grown in DMEM medium containing 10% bovine serum. Five days before the addition of the test compounds, the cells were washed by phosphate-buffered saline, and the medium was substituted with a phenol red-free DMEM medium containing 5% dextral charcoal-stripped FBS. The respective test compound was added in fresh medium and the control cell received only the vehicle (ethanol). The proliferation of MCF-7 was analyzed by the MTT assay, (3)H-TdR incorporation assay and flow cytometry. RESULTS: Compared with the ethanol control, the proliferation and DNA synthesis of the test cells treated with n-4-nonyphenol (8 x 10(-7) mol/L, 96 h), Bisphenol A (8 x 10(-7) mol/L, 96 h) or dibutylphthalate (32 x 10(-6) mol/L, 96 h) treatment was markedly enhanced in a time-dependent and dose-dependent manner. CONCLUSION: n-4-Nonyphenol, Bisphenol A and dibutylphthalate enhanced the proliferation of human breast cancer cell in vitro, which may demonstrate an estrogenic activity.

[Effects of genistein and zearalenone on proliferation of PEO4].

OBJECTIVE: The objective of this study was to investigate the estrogenic activity of genistein and zearalenone through their effects on the proliferative capacity of human ovarian PEO4. METHODS: Estrogen receptor-positive PEO4 cell was grown in DMEM medium containing 10% bovine serum. Five days before the addition of the test compounds, the cells were washed in phosphate-buffered saline, and the medium was substituted with a phenol red-free DMEM medium containing 5% dextran charcoal-stripped FBS. The respective test compound was added in fresh medium and the control cell received only the vehicle (ethanol). Cell proliferation was detected respectively by MTT assay, (3)H-TdR incorporation and flow cytometry. RESULTS: Compared with vehicle control, 96 x 10(-6) mol/L GS significantly inhibited PEO4 cell proliferation and DNA synthesis as measured by MTT and (3)H-TdR incorporation after treatment for 24 h. Alao, 32 x 10(-6) mol/L GS could exert inhibition on PEO4 cell growth as time extension to 48 h. 32 x 10(-6) mol/L approximately 96 x 10(-6) mol/L GS induced G(2)/M arrest. At low dose (< 8 x 10(-6) mol/L=, GS promoted proliferation in PEO4 cells. ZEA enhanced proliferation, promoted DNA synthesis and increased the S phase population in PEO4 cells. CONCLUSIONS: Genistein possess estrogenic activity and zearalenone have anti-estrogenic activity. They play different effects on the proliferation of human ovarian cancer cell. Genistein enhanced the proliferation of PEO4. Zearalenone inhibited its the proliferation. These results implied that genistein and zearalenone elicit different signal-transduction channel.

[Effects of environmental estrogens on apoptosis induced by estrogen depletion in T47D cells].

OBJECTIVE: To explore the effects of environmental estrogens (n-4-noniphenol, NP; bisphenol, BisA; and dibutylphthalate, DBP) on apoptosis induced by estrogen depletion in breast cancer T47D cells. METHODS: Human T47D breast cancer cells were grown in DMEM medium containing 10% bovine serum. Four days before adding the test compounds, the cells were washed in phosphate-buffered saline, and the medium was substituted with a phenol red-free DMEM medium containing 5% dextral charcoal-stripped FBS. Respective test compound was added in fresh medium and the control cell received only the vehicle (ethanol). Apoptotic features in T47D cell were analyzed by light microscope that was commonly used to define apoptosis. DNA integrity of T47D cells was examined by agarose gel electrophoresis. Hypodiploid population was detected by flow cytometry. RESULTS: The typical characters of apoptosis in T47D cells were observed after estrogen deletion and then disappeared following exposure to T47D cells at 32 x 10(-7) mol/L Np and 32 x 10(-7) mol/L BisA respectively. Inhibition of apoptosis at 32 x 10(-6) mol/L DBP was not shown in our study. CONCLUSION: N-4-noniphenol and Bisphenol A could inhibit apoptosis induced by estrogen deletion in breast cancer T47D cells. This result suggests that these environmental estrogens might involve in signal transduction connected with apoptosis.

Effect of zinc on bone metabolism in fetal mouse limb culture.

OBJECTIVE: To determine the effects of zinc-deficiency and zinc-excess on bone metabolism. METHODS: We developed the culture model of fetal mouse limbs (16th day) cultivated in self-made rotator with continuing flow of mixed gas for six days in vitro. The cultured limbs were examined by the techniques of 45Ca tracer and X-roentgenography. RESULTS: The right limbs cultivated had longer bone length, higher bone density than the left limbs uncultivated from the same embryo; and histologically, the right limbs had active bone cell differentiation, proliferation, increased bone trabecula, clearly calcified cartilage matrix, and osteogenic tissue. Compared with the control group, the zinc-deficient group and zinc-excess (Zn2+ 120 mumol/L) group contained less osteocalcin (BGP) and 45Ca content, and lower AKP activity; whereas zinc-normal (Zn2+ 45 mumol/L and Zn2+ 70 mumol/L) groups contained more BGP and 45Ca contents, and higher AKP (alkaline phosphatase) activity. CONCLUSION: Both zinc-deficiency and zinc-excess can alter bone growth and normal metabolism. The results indicate that the culture model of fetal mouse limbs (16th day) in vitro can be used as a research model of bone growth and development.