Methyltransferase inhibitor BIX01294 promotes the migration and inhibits decidualization of mouse uterine stromal cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of BIX01294 (BIX), a methyltransferase inhibitor, on the migration and decidualization of the stromal cells in mouse uterus.</p><p><b>METHODS</b>Mouse endometrial stromal cells were isolated and cultured from the uterus of pregnant mice on day 3.5 of gestation. The migration and decidualization of mouse endometrial stromal cells treated with BIX at different concentrations were observed with wound healing assay and real-time PCR.</p><p><b>RESULTS</b>The migration distance of mouse endometrial stromal cells increased as the BIX concentration increased within the range below 15 µmol/L. Compared with the control cells, the cells treated with BIX (15 µmol/L) showed significantly increased migration distances, but increasing BIX concentration to 20 µmol/L did not further increase the cell migration distance and began to cause cell death. Compared with the control cells, the BIX-treated stromal cells exhibited significantly down-regulated expression of Ehmt2 mRNA, and 15 µmol/L BIX caused inhibition of decidualization in the stromal cells.</p><p><b>CONCLUSION</b>Within a defined concentration range, BIX promotes the migration and inhibits decidualization of mouse uterine stromal cells by inhibiting the expression of Ehmt2 mRNA.</p>

Expression of FABP7 in mouse placenta tissue and human trophoblast HTR-8/Svneo cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect the expression of FABP7 in the placenta of pregnant mice and in HTR-8/Svneo cells.</p><p><b>METHODS</b>Real-time PCR and immunofluorescence were used to detect FABP7 mRNA and protein expressions in the uterine and placental tissue of pregnant mice at different days of gestation. FABP7 expression was also detected in cultured HTR-8/Svneo cells using immunofluorescence assay. The mice were treated with E, Por their combination for 6 and 24 h and Fabp7 mRNA level in the uterus was detected with real-time PCR.</p><p><b>RESULTS</b>At 7.5-10.5 days of gestation, the pregnant mice showed positive expressions of Fabp7 mRNA in the uterus and placenta, and FABP7 protein was detected in the decidualized cells and trophoblast giant cells. The expressions of FABP7 were detected at both the mRNA and protein levels in cultured HTR-8/Svneo cells. In mice treated with P4 alone or with E+Pfor 6 and 24 h, the expression level of Fabp7 mRNA was upregulated in the uterus. Fabp7 upregulation was observed in mice at 24 h following Etreatment but not at 6 h.</p><p><b>CONCLUSION</b>FABP7 is expressed in trophoblast giant cells and decidual cells in the placental tissue of mice and in cultured HTR-8/Svneo cells, suggesting the involvement of FABP7 in placental development and in maintenance of pregnancy. Eand Pcan regulate the expression of FABP7 in mouse uterus.</p>

Protective Effects of Curcumin against Sodium Arsenite-induced Ovarian Oxidative Injury in a Mouse Model / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Excessive reactive oxygen species (ROS) may lead to a number of reproductive diseases such as polycystic ovary syndrome. This study aimed to establish an animal model of ovarian oxidative stress and to assess the protective effect of curcumin against oxidative injury.</p><p><b>METHODS</b>Ovarian oxidative stress was induced in female Kunming mice (n = 40) with intraperitoneal injection of 8 mg/kg sodium arsenite (As) once every other day for 16 days; meanwhile, they were, respectively, treated by intragastric administration of 0, 100, 150, or 200 mg/kg (n = 10/group) curcumin once per day for 21 days. Ten normal mice were used as control. Then, the mice were injected intraperitoneally with BrdU and sacrificed; the right ovaries were collected for hematoxylin and eosin (HE) staining and BrdU immunohistochemistry, and the left ovaries for enzyme-linked immunosorbent assay (ELISA) and Western blotting analyses.</p><p><b>RESULTS</b>The ELISA results showed that ROS (11.74 ± 0.65 IU/mg in 8 mg/kg AS + 0 mg/kg curcumin group vs. 10.71 ± 0.91 IU/mg in control group, P= 0.021) and malondialdehyde (MDA) (0.32 ± 0.02 nmol/g in 8 mg/kg AS + 0 mg/kg curcumin group vs. 0.27 ± 0.02 nmol/g in control group, P= 0.048) increased while superoxide dismutase (SOD) (3.96 ± 0.36 U/mg in 8 mg/kg AS + 0 mg/kg curcumin group vs. 4.51 ± 0.70 U/mg in control group, P= 0.012) and glutathione peroxidase (17.36 ± 1.63 U/g in 8 mg/kg AS + 0 mg/kg curcumin group vs. 18.92 ± 1.80 U/g in control group, P= 0.045) decreased in the ovary after injection of As, indicating successful modeling of oxidative stress. Curcumin treatment could considerably increase SOD (4.57 ± 0.68, 4.49 ± 0.27, and 4.56 ± 0.25 U/mg in 100 mg/kg, 150 mg/kg, and 200 mg/kg curcumin group, respectively, allP < 0.05) while significantly reduce ROS (10.64 ± 1.38, 10.73 ± 0.71, and 10.67 ± 1.38 IU/mg in 100 mg/kg, 150 mg/kg, and 200 mg/kg curcumin group, respectively, allP < 0.05) and MDA (0.28 ± 0.02, 0.25 ± 0.03, and 0.27 ± 0.04 nmol/g in 100 mg/kg, 150 mg/kg, and 200 mg/kg curcumin group, respectively; bothP < 0.05) in the ovary. HE staining and BrdU immunohistochemistry of the ovarian tissues indicated the increased amount of atretic follicles (5.67 ± 0.81, 5.84 ± 0.98, and 5.72 ± 0.84 in 100 mg/kg, 150 mg/kg, and 200 mg/kg curcumin group, respectively, all P < 0.05), and the inhibited proliferation of granular cells under oxidative stress would be reversed by curcumin. Furthermore, the Western blotting of ovarian tissues showed that the p66Shc expression upregulated under oxidative stress would be lowered by curcumin.</p><p><b>CONCLUSION</b>Curcumin could alleviate arsenic-induced ovarian oxidative injury to a certain extent.</p>

Strategies of sperm collection from men with temporary penile erectile dysfunction on the day of oocyte retrieval and the outcomes of IVF-ET / 中华男科学杂志

<p><b>OBJECTIVE</b>To search for the optimal strategies for sperm collection from the patient with temporary penile erectile dysfunction (ED) on the day of oocyte pick-up ( OPU) in in vitro fertilization embryo transfer (IVF-ET).</p><p><b>METHODS</b>We retrospectively analyzed 93 cases of temporary ED on the OPU day of IVF-ET from January 2011 to May 2014, with fresh semen for 45 cases (group A), cryopreserved sperm before oocyte retrieval for 30 cases (group B), and frozen oocytes for 18 cases (group C). Group A was again subdivided into A1 (n = 18) and A2 n = 27) , the former intervened with oral sildenafil while the latter left untreated. We compared the rates of fertilization, high-quality embryo, and pregnancy among different groups.</p><p><b>RESULTS</b>No statistically significant differences were found among groups A, B and C in the age of the males and females, duration of infertility, numbers of obtained and mature oocytes, and rates of cleavage, or in the percentages of normal fertilization (80.78% vs 80.43% vs 84.77%), high-quality embryo (53.27% vs 52.97% vs 47.69%) and pregnancy (60.00% vs 56.77% vs 44.44%) (all P > 0.05). The rate of 3PN was markedly lower in group C (0.63%) than in A (9. 61%) and B (4.34%) (P < 0.05). There were no significant differences between groups A1 and A2 in the age of the males and females, duration of infertility, numbers of obtained and mature oocytes, and the rates of fertilization, cleavage, high-quality embryo, and pregnancy (all P > 0.05).</p><p><b>CONCLUSION</b>On the OPU day of IVF-ET, oral sildenafil can help temporary ED men to achieve penile erection and ejaculation without affecting the outcomes of assisted reproduction. Cryopreserved sperm can be used in case of predicted temporary ED and frozen oocytes can also be employed if sperm retrieval fails. However, to avoid puncture injury to the epididymis or testis, fresh semen should be the first choice.</p>

Curcumin inhibits endometriosis endometrial cells by reducing estradiol production

Endometriosis is a complex estrogen-dependent disease that is defined as the presence of endometrial gland and stroma outside the uterine cavity. Although the exact mechanism for the development of endometriosis remains unclear, there is a large body of research data and circumstantial evidence that suggests a crucial role of estrogen in the establishment and maintenance of this disease. This study is an attempt to assess the effect of curcumin on inhibiting endometriosis endometrial cells and to investigate whether such an effect is mediated by reducing estradiol production. Endometriotic stromal cells, normal endometrial stromal cells, endometriotic epithelial cells and normal endometrial epithelial cells were isolated and cultured. E[2] value of cells and the effect of curcumin on cell proliferation were evaluated. Finally, effect of curcumin on E[2] assay was detected. Electrochemiluminescence immunoassay results showed that E[2] value of endometriotic epithelial cells was higher than the endometriotic stromal cells [p=0.037], while the expression of E[2] in normal endometrial stromal and epithelial cells was extremely low. WST-8 result showed, compared with endometrial stromal cells, ectopic endometriotic stromal cells had a higher growth rate. After intervene with curcumin [10micromol/L, 30micromol/L and 50micro mol/L] for 0-96h, the number of endometriotic stromal cells was reduced and cells growth slowed, compared with 0micromol/L group. Compared with 0micromol/L group, E[2] level was lower after treatment with curcumin, especially in 30micromol/L and 50micromol/L group. In summary, in this study we found that E[2] is important in ectopic endometrium, and epithelial cell is in dominant position with E[2] secretion. Curcumin was able to suppress the proliferation of endometrial cells by reducing the E2 value.

Association between single nucleotide polymorphisms of 5'-untranslated region of GPx4 gene and male infertility / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the association between the single nucleotide polymorphisms (SNPs) of the 5'-untranslated region (5'-UTR) of phospholipid hydroperoxide glutathione peroxidase (GPx4 or PHGPx) gene and oligo- or asthenozoospermic male infertility.</p><p><b>METHODS</b>The 5'-UTR region of the GPx4 gene was amplified from infertile men and controls using the polymerase chain reaction and was analyzed for polymorphisms by direct sequencing.</p><p><b>RESULTS</b>A total of 9 SNPs were present in the cohort, however there were no significant differences in these 9 SNPs between the case and control groups. According to the results of linkage disequilibrium analysis and haplotype construction, one haplotype (rs757229-rs757230-rs4588110-rs3746165-rs3746166: C-G-G-T-A) was present only in the control men, and significant difference was detected(P< 0.01).</p><p><b>CONCLUSION</b>The SNPs of 5'-UTR region of the GPx4 gene might not be associated with oligo- or asthenozoospermic male infertility. However, the haplotype (rs757229-rs757230-rs4588110- rs3746165-rs3746166: C-G-G-T-A) might be a protective haplotype.</p>

Establishment of immortalized cell lines and genetic stability evaluation for a family with spinocerebellar ataxia type 2 / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>Immortalized cell lines of spinocerebellar ataxia type 2 (SCA2) with Parkinson disease symptoms were established in order to provide experimental material for future study.</p><p><b>METHODS</b>The immortalized cell lines were constructed by using Epstein Barr virus and cyclosporine A. Microsatellite markers were detected to see whether there is any change between the cell lines and the original blood samples, and the genetic stability of the cell lines were evaluated.</p><p><b>RESULTS</b>Twenty-five immortalized cell lines were established successfully from the family and the microsatellite markers were unchanged.</p><p><b>CONCLUSION</b>The karyotypes of the immortal cell lines were normal and the cell lines were genetically stable.</p>

Collection of a Chinese pedigree with Parkinson's disease and linkage analysis of nine susceptibility genes / 中国医学科学院学报

<p><b>OBJECTIVE</b>To analyze the susceptibility genes of a Parkinson's Disease (PD) family.</p><p><b>METHODS</b>The blood samples of a four-generation classic idiopathic PD family were collected. Two-point LOD score method was applied to analyze the linkage disequilibrium between the disease locus and microsatellite markers.</p><p><b>RESULTS</b>We studied 13 markers near the 9 genes that had been reported to be associated with PD. No obvious evidence showed that the selected markers had anything correlation with PD locus.</p><p><b>CONCLUSION</b>These 9 genes are not the susceptibility genes of PD in this family.</p>