Maternal vitamin B1 is a determinant for the fate of primordial follicle formation in offspring.

The mediation of maternal-embryonic cross-talk via nutrition and metabolism impacts greatly on offspring health. However, the underlying key interfaces remain elusive. Here, we determined that maternal high-fat diet during pregnancy in mice impaired preservation of the ovarian primordial follicle pool in female offspring, which was concomitant with mitochondrial dysfunction of germ cells. Furthermore, this occurred through a reduction in maternal gut microbiota-related vitamin B1 while the defects were restored via vitamin B1 supplementation. Intriguingly, vitamin B1 promoted acetyl-CoA metabolism in offspring ovaries, contributing to histone acetylation and chromatin accessibility at the promoters of cell cycle-related genes, enhancement of mitochondrial function, and improvement of granulosa cell proliferation. In humans, vitamin B1 is downregulated in the serum of women with gestational diabetes mellitus. In this work, these findings uncover the role of the non-gamete transmission of maternal high-fat diet in influencing offspring oogenic fate. Vitamin B1 could be a promising therapeutic approach for protecting offspring health.

[Relationship of S100B protein expression and the pathogenesis of early-onset and late-onset preeclampsia].

OBJECTIVE: To investigate the relationship of S100B protein expression and the pathogenesis of early-onset and late-onset preeclampsia. METHODS: Sixty patients with preeclampsia who received caesarean section at Qingdao Municipal Hospital from October 2010 to September 2011 were enrolled in this study. Thirty cases were early-onset preeclampsia (referred as early-onset preeclampsia group, < 34 weeks), and the other 30 cases were late-onset preeclampsia (referred as late-onset preeclampsia group, ≥ 34 weeks). Thirty women who received caesarean section because of pelvic structural deformities, breech presentation, macrosomia and social factors were included as the control group. The expression of S100B mRNA in the placenta was detected by reverse transcription (RT)-PCR. The expression of S100B protein in the placenta was detected by immunohistochemistry. RESULTS: (1) S100B mRNA was expressed in the trophoblasts of preeclampsia and control groups. The expression of S100B mRNA in early-onset preeclampsia group (0.73 ± 0.11) was significantly higher than the control group (0.58 ± 0.08) and late-onset preeclampsia group (0.64 ± 0.10, P < 0.05). There was no significant difference between late-onset preeclampsia group and the control group (P > 0.05). (2) S100B protein was expressed in the plasma membrane and cytoplasm of the trophoblasts, correlated positively with the brownish yellow and brown particles inside the cells. It was expressed in all the three groups. Immunohistochemistry revealed that the expression of S100B protein in the placenta of early-onset preeclampsia group was 100% (30/30), significantly higher than those of late-onset preeclampsia group and the control group, in which the positive rate were 70% (21/30) and 63% (19/30) respectively (P < 0.05). There was no difference between late-onset preeclampsia group and the control group (P > 0.05). CONCLUSION: Early-onset and late-onset preeclampsia may have different etiology and pathogenesis. S100B may be a factor in the pathogenesis of early-onset preeclampsia.

[Effects of transient receptor potential V6 silence on proliferation and apoptosis of trophoblasts].

OBJECTIVE: To investigate the effects of the transient receptor potential V6 (TRPV6) gene silencing on the proliferation and apoptosis of trophoblasts HTR-8/SVneo cells. METHODS: siRNA sequences targeting the TRPV6 gene were constructed and then transfected into HTR-8/SVneo cells mediated by liposome. The cells were divided three groups, including blank control (add the reagent of transfenction), negative control groups (transfecting nonspecific siRNA) and experimental groups (transfecting TRPV6-siRNA). Those cells in every group were collected at 24, 48, 72 hours after transfecting. The expression levels of TRPV6 mRNA were detected by reverse transcription (RT) PCR at different times after transfecting. The effects of siRNA on the proliferation and apoptosis of the cells were assayed by methyl thiagolyl tetragolium (MTT) and flow cytometry at different times after transfecting. RESULTS: siRNA TRPV6 transfection could inhibit the expression of TRPV6 mRNA in the HTR-8/SVneo cells. The expression was decreased with the extension of time, by 0.72 ± 0.02, 0.54 ± 0.02 and 0.29 ± 0.01 after 12, 48 and 72 hours of siRNA transfection as compared with the blank control and the negative control groups (P < 0.01). The rates of proliferation inhibition were (19.29 ± 1.23)%, (32.12 ± 1.35)% and (46.51 ± 1.42)% at 24, 48 and 72 hours respectively when compared with the blank control (2.12 ± 0.03)%, (2.42 ± 0.02)%, (3.13 ± 0.04)% and the negative control groups (2.37 ± 0.01)%, (2.61 ± 0.05)%, (2.93 ± 0.03)% (P < 0.01). The apoptosis rates of HTR-8/SVneo cells was 16.21% at 48 hours after transfected with siRNA TRPV6, which were significantly higher than 3.27% in the blank control and 5.34% in the negative control groups (P < 0.05). CONCLUSION: Silenceing of TRPV6 genen could inhibit the proliferation and increase the apoptosis of extravillous trophoblas of human placenta.

Altered expression of claudin-3 and claudin-4 in ectopic endometrium of women with endometriosis.

OBJECTIVE: To investigate the expression of claudin-3 and claudin-4 in the eutopic and ectopic endometrium of women with endometriosis and to evaluate the role of claudin-3 and claudin-4 in the pathogenesis of endometriosis. DESIGN: Cross-sectional measurement of gene expression levels of claudin-3 and claudin-4 on endometriotic tissue. SETTING: Academic. PATIENT(S): Thirty-five patients with endometriosis and 35 healthy women who were free of endometriosis were recruited for the study. INTERVENTION(S): Expression of claudin-3 and claudin-4 were investigated with immunohistochemical analysis, Western blot, and real-time polymerase chain reaction. Morphologic change of tight junction was also observed in different kinds of endometria. MAIN OUTCOME MEASURE(S): The expression levels of claudin-3 and claudin-4 in epithelial cells from 35 ectopic endometrial tissues, 27 eutopic endometrial tissues from women with endometriosis, and 35 normal endometrial tissues from women without endometriosis. RESULT(S): Expression of claudin-3 and claudin-4 was significantly lower in the ectopic endometriotic tissue than in the eutopic endometrium from women with endometriosis and normal controls at both the messenger RNA and protein levels. No significant difference was found between eutopic endometrium from women with endometriosis and normal endometrium from women without endometriosis. CONCLUSION(S): Down-regulated expression of claudin-3 and claudin-4 in ectopic endometrium suggests that claudin-3 and claudin-4 might play a pathogenic role in the formation of endometriosis.

[Expression of claudin-4 in eutopic and ectopic endometrium of women with endometriosis].

OBJECTIVE: To investigate the expression of claudin-4 in the eutopic and ectopic endometrium of women with endometriosis and evaluate the role of claudin-4 in the pathogenesis of endometriosis. METHODS: Thirty-five women with endometriosis and 35 controls were studied. Expression of claudin-4 was investigated using immunohistochemistry, western blot and RT-PCR, respectively. Morphologic change of tight junction was also observed in different kinds of endometria RESULTS: (1) Glandular epithelial cells of control endometrium and eutopic endometrium showed intact tight junctions in electron micrographs, whereas the morphology of tight junctions in ovarian endometriotic tissue was disrupted and collagen bundles could be easily detected. (2) The immunohistochemical staining of claudin-4 was localized to the glandular epithelial cell membrane. Deficient or weak staining was found in ovarian endometriotic tissues. In control endometrium, eutopic and ectopic endometrium of women with endometriosis, the expression of claudin-4 protein was 89 +/- 24, 84 +/- 22 and 27 +/- 14, respectively. Relative expression of claudin-4 mRNA was 14.5 +/- 6.8, 13.8 +/- 9.5 and 2.6 +/- 2.5, respectively. Expression of claudin-4 was significantly lower in the ectopic endometriotic tissue than in the eutopic endometrium and the control at both mRNA and protein levels (P<0.05). No significant difference was found between eutopic endometrium from women with endometriosis and control endometrium from women without endometriosis (P>0.05). CONCLUSION: Down-regulated expression of claudin-4 might play a pathogenic role in the formation of endometriosis.

[Influence of 4-aminopyridine on cell proliferation through inhibiting voltage-gated K+ channel in ovarian cancer].

OBJECTIVE: To study the inhibition of cell proliferation by 4-aminopyridine (4-AP) in the human ovarian cancer cell SKOV3. METHODS: The expression of voltage-gated K(+) channel on the SKOV3 cell line was detected by reverse transcription polymerase chain reaction. Inhibition of voltage-gated K(+) current by 4-AP through the whole-cell patch-clamp technique on SKOV3 cell line was recorded. The influence on the cell-cycle of the SKOV3 cell line by 4-AP was observed by flow cytometry and the inhibition of the cell proliferation was studied using methyl thiazolyl tetrazolium (MTT) method. RESULTS: Voltage-gated K(+) channel was expressed in SKOV3 cell. Exposure of the SKOV3 cell to 5 mmol/L 4-AP reduced K(+) current by 52.5%. The 4-AP at 5 mmol/L significantly effected the progression of cell cycle with a 72 h treatment of SKOV3 cell. Exponentially growing cells without inhibitors had a distribution of 39.7% in G(0)/G(1) phase, 57.3% in S phase, and 3.0% in G(2)/M phase. In 4-AP-treated cells, the proportion of G(0)/G(1) cells increased significantly to 62.3% (P < 0.05), while there was a significant decrease in S phase cells (36.2%, P < 0.05) and G(2)/M phase (1.4%, P < 0.05). Incubation of the SKOV3 cells with 0.1, 1, 5, 10, 15, 20 mmol/L 4-AP resulted in a concentration-dependent reduction in the number of viable cells as compared with the control, and the inhibition rate was 17.5%, 35.0%, 54.6%, 69.1%, 71.2%, 72.8% respectively (vs control, P < 0.01). CONCLUSION: The voltage-gated K(+) channels expressed by SKOV3 play an important role in SKOV3 cell proliferation.