Mechanism of cleft palate in C57BL/6N mice induced by retinoic acid / 中华口腔医学杂志

Objective@#To investigate the mechanism of cleft palate in mice induced by excessive all-trans retinoic acid (atRA).@*Methods@#The pregnant mice were randomly divided into atRA-treated group (n=27) and control group (n=27). atRA-treated group was given by gavage a single dose of atRA (100 mg/kg) at 8: 00 AM on gestation day 10 (GD10) and the control group was given by gavage the isopyknic corn oil. At GD13-GD15, the fetal mice palate development was observed by HE staining. The mouse embryonic palatal mesenchymal cell proliferation was detected by 5-bromo-2-deoxyuridine (BrdU) immunohistochemistry. The expressions of Smad2, phospho-Smad2 (p-Smad2), Smad4 and Smad7 in mouse embryonic palatal mesenchyme were analyzed by Western blotting.@*Results@#At GD13-GD15, compared with the control, the ratio of BrdU-positive cells in the palatal mesenchyme of atRA-treated fetuses decreased significantly (P<0.05), especially at GD14, atRA inhibition rate was (65.4±1.7)%. Moreover, atRA decreased the levels of p-Smad2 and Smad4 in embryonic palate mesenchymal cells, whereas the expression of Smad7 was significantly increased at GD14 and GD15.@*Conclusions@#atRA may lead to cleft palate by inhibiting the activation of Smad signaling pathway and affecting the proliferation of palatal mesenchymal cells.

Association between environmental dioxin-related toxicants exposure and adverse pregnancy outcome: systematic review and meta-analysis

Dioxin-related compounds are associated with teratogenic and mutagenic risks in laboratory animals, and result in adverse pregnancy outcomes. However, there were inconsistent results in epidemiology studies. In view of this difference, we conducted a systematic review and meta-analysis to examine this association and to assess the heterogeneity among studies. Comprehensive literature searches were performed to search for relevant articles published in English up to 15 May 2012. In total, we identified 15 studies which included 9 cohort and 6 case control studies. The Cochrane Q test and index of heterogeneity [I2] were used to evaluate heterogeneity. In either cohort studies [I2=0.89, p<0.0001] or case control studies [I2=0.69, p=0.02], significant heterogeneity of risk estimates were observed. Subgroup analyses found no significant increased risk of adverse pregnancy outcome with air dioxin-related compounds exposure [RR=0.99, 95% CI:0.85-1.16], no significant increased risk of spontaneous abortion [SAB] with exposure to food dioxin-related compounds [RR=1.05, 95% CI:0.80-1.37], higher significant risks of low birth weight [LBW] with exposure to food dioxin-related compounds [RR=1.55, 95% CI:1.24-1.94], and higher significant risks of birth defects with maternal solid contaminants dioxin exposure [OR=1.24, 95% CI:1.19-1.29]. In conclusion, more evidences are needed to confirm the association between environmental dioxin-related compounds exposure and pregnancy outcome

Association between maternal MTHFR polymorphisms and nonsyndromic cleft lip with or without cleft palate in offspring, a meta-analysis based on 15 case-control studies

The methylenetetrahydrofolate reductase [MTHFR] is thought to be involved in the development of nonsyndromic cleft lip with or without cleft palate [NSCL/P]. However, conflicting results have been obtained when evaluating the association between maternal MTHFR C677T and A1298C polymorphisms and the risk of NSCL/P. In light of this gap, a meta-analysis of all eligible case-control studies was conducted in the present study. A total of 15 case-control studies were ultimately identified after a comprehensive literature search and Hardy-Weinberg equilibrium [HWE] examination. Cochrane's Q test and index of heterogeneity [I[2]] indicated no obvious heterogeneity among studies. Fixed or random-effects models were used to calculate the pooled odds ratios [ORs]. The results showed that the TT genotype in mothers increased the likelihood of having NSCL/P offspring 1.25 times [95% CI: 1.047-1.494] more than the CC homozygotes. Meanwhile, maternal TT genotype increased the risk of producing NSCL/P offspring in recessive model [OR=1.325, 95% CI: 1.124-1.562]. However, the CT heterozygote and the CT+TT dominant models had no association with NSCL/P offspring compared with the CC wild-type homozygote model. Subgroup analyses based on ethnicity indicated that maternal TT genotype increased the likelihood of having NSCL/P offspring in Whites [OR=1.308, 95% CI: 1.059-1.617] and Asians [OR=1.726, 95% CI: 1.090-2.733] in recessive model. Also, subgroup analyses based on source of control showed that mothers with the 677TT genotype had a significantly increased susceptibility of having NSCL/P children in hospital based population [HB] when compared with CC homozygotes [OR=1.248, 95% CI: 1.024-1.520] and under the recessive model [OR=1.324, 95% CI: 1.104-1.588]. Furthermore, maternal A1298C polymorphism had no significant association with producing NSCL/P offspring [dominant model OR=0.952, 95% CI: 0.816-1.111, recessive model OR=0.766, 95% CI: 0.567-1.036]. MTHFR C677T polymorphism is associated with the risk of generating NSCL/P offspring, and being a 677TT homozygote is a risk factor. MTHFR A1298C polymorphism was not associated with generating NSCL/P offspring. However, further work should be performed to confirm these findings

STUDY OF THE COMBINED PREVENTIVE EFFECT OF FOLIC ACID AND VITAMIN B_(12) ON THE DEVELOPMENTAL TOXICITY OF ALCOHOL / 营养学报

Objective: To investigate the combined preventive effect of folic acid (FA) and vitamin B12 (VB12) on the developmental toxicity of alcohol. Methods: To build animal model with the developmental toxicity by giving 5g/kg bw alcohol (25% ethanol) intragastrically (IG) to ICR mice during gestational day (GD) 6-15. In addition to alcohol, three groups were given FA 60 mg/kg bw, VB12 1 mg/kg bw, FA 60 mg/kg bw + VB12 1 mg/kg bw respectively by IG during GD1-GD16. An alcohol model group and a negative control group were set. All dams were killed at GD18. Results: Compared to the alcohol model group, the pregnant mice of FA + VB12 combined intervention group put on more weight during pregnancy; the live fetal rate; the fetal weight, body length and tail length were all increased; the abnormal ossification rate of sternum, occipital bone, and four limbs dropped (P

Effects of genistein on proliferation of osteoblast of post-menopausal patients and the likely molecular mechanisms / 中华老年医学杂志

Objective This study was designed to explore the effects of phytoestrogen genistein on proliferation of post-menopausal osteoblast and the likely molecular mechanisms. Methods Osteoblast cultures were prepared from the upper femur of postmenopausal patients. Osteoblast proliferation was determined by Methyl Thiazolyl Tetrazolium(MTT)assay and cell cycle distribution by cytometry. The protein expressions of cyclin D and E were examined using Western-blot. Results Genistein (10~(-8) to 10~(-6) mol/L) stimulated cell viability of postmenopausal osteoblasts and increased the distribution ratio of the cells at the G2/M and S phases (P

Effects of zinc on bone development in mouse embryo in vitro / 中华预防医学杂志

<p><b>OBJECTIVE</b>This study was designed to explore the effects of zinc on bone development.</p><p><b>METHODS</b>Forelimbs of mice with 16 d gestation were cultured by a rotating device.</p><p><b>RESULTS</b>Contents of OC and (45)Ca and activities of AKP in the bone tissues cultured at zinc-deficiency media and at media with 120 micro mol/L of Zn(2+) decreased significantly. Synthesis of OC, absorption of calcium and activities of AKP in the bone tissues cultures at media with 45 and 70 micro mol/L of Zn(2+) increased significantly. Radiograph of bone tissues showed that length of long bone cultured at zinc-deficiency media and at media with 120 micro mol/L of Zn(2+) shortened and their density reduced, and those cultured at media with 45 and 70 micro mol/L of zinc increased, as compared with self-control bone. Histological analysis showed the death of bone cells and loss of stroma in the bone tissues cultured at media with 120 micro mol/L of Zn(2+), and active proliferation and differentiation of bone cells, and secretion and synthesis of osteoid increased in the bone tissues cultured at media with 45 and 70 micro mol/L of zinc.</p><p><b>CONCLUSIONS</b>Adequate supplementation of zinc could promote formation and development of bone tissues and deficiency or excess of zinc could alter their growth and development and normal metabolism.</p>

Effects of Environmental Estrogens on Apoptosis of Ovarian Cancerous PEO4 Cells / 环境与健康杂志

Objective To study the effects of dietary estrogens genistein(GS)and zearalenone(ZEA)on apoptosis in-duced by estrogen depletion in PEO4cells.Methods The monolayer ovarian cancer cell line PEO4cells were cultured in DMEM medium containing10%bovine serum.Before the addition of the testing compounds the cells were washed in phosphate-buffered saline and the medium was displaced with a phenol red-free DMEM medium containing5%dextral charcoal-stripped FBS and the cells were cultured for5days in order to exhaust the estrogen stored in the cells,and then cells were divided into5groups,including solvent control group,estrogen control group,anti-estrogen control group and2experimental groups.After treatment the apoptotic features of the cells were observed by cellular morphology,DNA fragmentation and location and height of cell hypodiploid were indicated by flow cytometry.Results The typical characteristics of apoptosis in PEO4cells were observed after estrogen deletion and then disappeared following exposure of the PEO4cells to32?10 -9 mol/L and96?10 -9 mol/L ZEA for72hrs.32?10 -6 mol/L and96?10 -6 mol/L GS could significantly aggravate apoptosis in PEO4cells.Conclusion Zearalenone is a kind of mycoestrogen that has estrogenic activity to inhibit apoptosis in PEO4cells.Genistein is a kind of phytoestrogen that has anti-estrogen activity(tamoxifen-like)to promote apoptosis in PEO4cells with the high doses range.

ISOFLAVONES INDUCED APOPTOSIS IN PROSTATE CANCER CELL PC-3 / 营养学报

Objective: To explore the mechanisms of the effect of isoflavone in reducing prostate cancer incidence throught studying the effects of isoflavone on apoptosis in PC-3 cell. Methods: Prostate cancer cell PC-3 was grown in RPMI 1640 medium supplemented with 10% bovine serum and 10 000 U/ml of penicillin/streptomycin in an incubator maintained at 5% CO 2 95% air and 100% humidity at 37 ℃. The respective test compound was added in fresh medium and the control cell received only the vehicle (MDSO). Apoptosis of PC-3 cells was analyzed by cell morphology under light microscope, agarose gel electrophoresis and flow cytometry. Results: Exposure of PC-3 cell to 50 ?mol/L GS, 75 ?mol/L DA and 75 ?mol/L GL after 72 h, the cell morphology indicated typical features commonly used to define apoptosis; agarose gel electrophoresis demonstrated laddered electrophoretic profiles of oligonucleosomal DNA fragments indicative of apoptosis, and flow cytometric analysis revealed a hyperdiploid population in the tested cells. Conclusion: Isoflavones could induce apoptosis in prostate cancer cells PC-3 and it may be the main cause of their role in cancer inhibition.

EFFECT OF PHYTOESTROGENS ON THE PROLIERATION OF MCF-7 IN VITRO / 营养学报

8 ?mol/L)was markedly restrained. Zearalenone, like estradiol, could markedly increase the proliferation of MCF-7 cells. The effects were time-dependent and dose-dependent. Conclusion: The tested phytoestrogens are biologically active, and they can differently affect the proliferation of human breast cancer cells in vitro. These data suggest that genistein may have preventive and therapeutic applications against breast tumors.

IN VITRO STUDY ON EFFECTS OF ZINC ON BONE DEVELOPMENT / 营养学报

Objective: To explore the effects of zinc on bone development in vitro. Methods: Mice of 16 gestation were killed by snapping cervical vertebra, and the front limbs were cut off and cultured in basic medium. Which contained 20 ?mol/L Zn 2+ (control group). In Zn deprived group, the basic medium was added TPEN to final concentration 20 ?mol/L. In other three groups, the basic medium was added ZnSO 4 to final concentration of Zn 2+ at 45, 70, and 120 ?mol/L respectively. Results: 1. Compared with control limb(uncultivated),the test limb(cultivated for 6 d)showed longer bone length and higher bone density.Histological analysis showed active bone cell differentiation, proliferation, increased bone trabecula.2.Compared with control group, the Zn-deprived and Zn 2+ 120 ?mol/L group expressed decreased osteocalcin (OC) and 45 Ca content and lower activity of AKP; Zn 2+ 45 ?mol/L and 70 ?mol/L group presented increased OC and 45 Ca content and higher activity of AKP.3.Compared with the control group, Zn-deprived and Zn 2+ 120?mol/L group showed shorter bone length and lower bone density; Zn 2+ 45 ?mol/L and 70?mol/L group showed longer bone length and higher bone density. Conclusion: Zinc is involved in bone metabolism and growth; both zinc-deficiency and zinc-excess can alter bone growth and normal metabolism.

EFFECTS OF ALL-TRANS RETINOIC ACID ON PALATAL FUSION AND ITS MOLECULAR MECHANISMS / 营养学报

Objective To investigate the effect of excessive all-trans retinoic acid (atRA) on mouse embryonic palatal fusion and the mechanism. Method Palatal shelves from embryonic D 13 embryonic mice were cultured in BGJb medium and treated with vehicle control only or 5 ?mol/L atRA for 72 h. Palatal fusion was examined by hemagglutinin esterase. Apoptosis and laminin were detected by TUNEL and immunohistochemistry, respectively. The level of Smad2 phosphorylation (pSmad2) was analyzed by Western blot. Results atRA led to failure of palatal fusion and inhibited the migration and apoptosis of medial edge epithelial cells (MEE) and degradation of basal lamina within, compared with control palatal shelves in cultures. Additionally, apoptosis was detected in mesenchyme of atRA-treated palatal shelves. Further experiment revealed that pSmad2 was abrogated by atRA. Conclusion atRA induced failure of palatal fusion through inhibition of apoptosis of the MEE cell and degradation of basal lamina within medial edge epithelial seam. Inhibition of pSmad2 may account for the failure of palatal fusion by atRA.

EFFECTS OF GENISTEIN ON TUMOR-ASSOCIATED GENE EXPRESSION IN MCF-7 CELLS / 营养学报

Objective: To characterize the effects of genistein (GS) on tumor-associated gene expression in MCF-7 cells and to explore the mechanisms. Method: MCF-7 cells were treated with GS (75?10-6 mol/L) or with vehicle (0.1% ethanol, EtOH) for 72 h. The cells were collected and total RNA was extracted, marked by two different fluorescence dyes (Cy3 and Cy5) using reverse transcriptional reaction, respectively. The derived cRNA was hybridized to human tumor-associated microarrays. Data were processed using the Scan Array 3000 and ontological analysis was performed using the Ima Gene 3.0 software. Results: At 72 h, 18 transcripts were downregulated compared to EtOH vehicle control by greater than 2- fold and 4 were upregulated by less than 0.5-fold. The predominant ontological groupings were oncogenes, cell proliferation, apoptosis, and estrogen-regulation genes. Conclusion: GS treatment is associated with significant changes in gene expression in several functional categories in MCF-7 cells, and this might account for the role of genistein in prevention of breast cancer.

THE MECHANISMS OF PROLIFERATION AND APOPTOSIS IN MCF-7 CELLS BY DIETARY ESTROGENS / 营养学报

Objective: This study was designed to investigate the molecular mechanisms of proliferation and apoptosis by genistein and zearalenone through regulation of mRNA and protein expression of PCNA, bcl-2 and bax in breast cancer MCF-7 cells. Methods: The cells were maintained in DMEM medium with 10% fetal bovine serum. Five days before the beginning of experiments, the cells were seeded in phenol red-free DMEM medium containing 5% charcoal dextran–treated FBS. The cells were harvested and seeded in 6-well culture plates or in 75 ml flacks. After various concentrations of genistein and zearalenone treatments for 72 h, the cells were harvested and mRNA and protein expression of proliferation cell nuclear antigen(PCNA), bcl-2 and bax were detected by reverse transcription-polymerase chain reaction and immunohistochemistry, respectively. Results: At the concentration of 75 ?mol/L, GS could significantly down-regulate bcl-2 and PCNA mRNA expression and up-regulate bax mRNA expression, and zearalenone indicated an opposite result. These results were further confirmed by following immunohistochemistry. Conclusion:PCNA, bcl-2 and bax pathway might be involved in cell proliferation and apoptosis events regulated by dietary estrogens genistein and zearalenone in breast cancer MCF-7 cells.

EFFECTS OF GENISTEIN ON APOPTOSIS IN HUMAN OSTEOBLAST CELLS / 营养学报

Objective:To investigate the effects of genistein (Gen) on apoptosis in human osteoblast. Methods:Human periosteum was maintained in DMEM medium containing 10% newborn calf serum through four passages to isolate and proliferate osteoblast. Then, cells were maintained in phenol red-free DMEM supplemented with 5% charcoal-stripped FCS for 4 d before addition of treatments. Flow cytometer and DNA fragment were used to explore apoptosis in osteoblast, and immunohistochemistry was used to study protein expression of Bax, Bcl-2 and caspase-3. Results:Estrogen depletion could induce apoptosis in human osteoblast. With the similarity of 10-8 mol/L E2, at the conentrations of 10-6 mol/L and 10-5 mol/L, Gen could inhibit apoptosis induced by estrogen depletion. Compared with vehicle control, Gen significantly decreased protein expression of Bax and caspase-3 and increased protein expression of Bcl-2. Conclusion:Genistein could inhibit apoptosis induced by estrogen depletion in human osteoblast.