Clinical data-based modeling of IVF live birth outcome and its application.

BACKGROUND: The low live birth rate and difficult decision-making of the in vitro fertilization (IVF) treatment regimen bring great trouble to patients and clinicians. Based on the retrospective clinical data of patients undergoing the IVF cycle, this study aims to establish classification models for predicting live birth outcome (LBO) with machine learning methods. METHODS: The historical data of a total of 1405 patients undergoing IVF cycle were first collected and then analyzed by univariate and multivariate analysis. The statistically significant factors were identified and taken as input to build the artificial neural network (ANN) model and supporting vector machine (SVM) model for predicting the LBO. By comparing the model performance, the one with better results was selected as the final prediction model and applied in real clinical applications. RESULTS: Univariate and multivariate analysis shows that 7 factors were closely related to the LBO (with P < 0.05): Age, ovarian sensitivity index (OSI), controlled ovarian stimulation (COS) treatment regimen, Gn starting dose, endometrial thickness on human chorionic gonadotrophin (HCG) day, Progesterone (P) value on HCG day, and embryo transfer strategy. By taking the 7 factors as input, the ANN-based and SVM-based LBO models were established, yielding good prediction performance. Compared with the ANN model, the SVM model performs much better and was selected as the final model for the LBO prediction. In real clinical applications, the proposed ANN-based LBO model can predict the LBO with good performance and recommend the embryo transfer strategy of potential good LBO. CONCLUSIONS: The proposed model involving all essential IVF treatment factors can accurately predict LBO. It can provide objective and scientific assistance to clinicians for customizing the IVF treatment strategy like the embryo transfer strategy.

Upregulated long noncoding RNA Linc00261 in pre-eclampsia and its effect on trophoblast invasion and migration via regulating miR-558/TIMP4 signaling pathway.

Pre-eclampsia (PE) is a leading cause of maternal and perinatal morbidity and mortality but the exact underlying mechanisms of PE pathogenesis remain elusive. Accumulated data suggested that the long noncoding RNAs (lncRNAs) play important roles in the pathogenesis of PE. The present study identified the changes of lncRNA Linc00261 in PE and its effects on trophoblasts invasion and migration. Our results showed that the expression of Linc00261 was upregulated in placental tissues of PE women compared with those of healthy pregnant women. Overexpression of Linc00261 suppressed cell invasion and migration, induced cell apoptosis, and caused cell-cycle arrest at G0 /G1 phase of HTR-8/SVneo cells; while knockdown of Linc00261 had the opposite effects on the HTR-8/SVneo cells. Mechanistic studies showed Linc00261 functioned as a competing endogenous RNA for miR-558 in HTR-8/SVneo cells, and miR-558 was negatively regulated by Linc00261. The expression level of miR-558 in the PE group was significantly lower than the control group, and the expression level of Linc00261 was negatively correlated with the expression level of miR-558 in the placental tissues of women with PE. Furthermore, miR-558 was found to negatively regulate the expression of TIMP metallopeptidase inhibitor 4 (TIMP4) via targeting the 3' untranslated region in the HTR-8/SVneo cells. Overexpression of miR-558 increased HTR-8/SVneo cell invasion and migration, which was attenuated by TIMP4 overexpression. More importantly, both overexpression of miR-558 and knockdown of TIMP4 partially reversed the suppressive effects of Linc00261 overexpression on cell invasion and migration of HTR-8/SVneo cells. Collectively, our results for the first time showed the upregulation of Linc00261 in the placental tissues of severe PE patients. The mechanistic results indicated that Linc00261 exerted the suppressive effects on the trophoblast invasion and migration via targeting miR-558/TIMP4 axis, which may involve in the pathogenesis of PE.

Transcriptomic changes and potential regulatory mechanism of intrauterine human chorionic gonadotropin co-cultured with peripheral blood mononuclear cells infusion in mice with embryonic implantation dysfunction.

BACKGROUND: This study aimed to explore whether intrauterine infusion of peripheral blood mononuclear cells (PBMCs) could induce favorable transcriptomic changes in the endometrium for embryo implantation and the potential mechanism. METHODS: Twenty-one mice were randomly divided to five groups, including a normal pregnancy (NP) group, an embryo implantation dysfunction (EID) group, an EID with human chorionic gonadotropin (hCG) group, an EID with PBMCs group, and an EID with hCG co-cultured with PBMCs group. The endometrium in the implantation window from mice were collected and determined by RNA sequencing (RNA-Seq), and the expression of significantly different genes with high degree of coincidence was recommended and validated by quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: There were totally 1,366 up-regulated and 1,374 down-regulated genes in the EID mice compared with the normal pregnant mice. We selected (fold change ≥2, P<0.05) and verified the candidate genes associated with embryo implantation, immune response and other reproductive processes in previous reports by qRT-PCR. Leukemia inhibitory factor (LIF), solute carrier family 15 member 2 (SLC15A2), retinoic acid receptor responder 1 (RARRES1), vascular cell adhesion molecule 1 (VCAM1) were down-regulated and musculin (MSC), chemokine (C-X-C motif) ligand 14 (CXCL14) were up-regulated significantly in EID group (P<0.05), and the synergistic effects of hCG were seen. In addition, the expression of glucocorticoid receptor (GR)-ß in PBMCs of NP mice was higher than that of EID mice, and up-regulated GR-ß in EID mice could significantly increase the expression of LIF, SLC15A2, RARRES1 and VCAM1, and decrease the expression of CXCL14 and MSC, which indicated GR-ß might be a transcriptional factor of the six genes above. CONCLUSIONS: Intrauterine PBMCs perfusion might improve the performance of impaired endometrial receptivity by regulating LIF, SLC15A2, RARRES1, VCAM1, MSC as well as CXCL14, and hCG could enhance the effect of PBMCs. In addition, GR-ß, as a transcriptional factor, could regulate the six genes in PBMCs.

Effect of immunotherapy on patients with unexplained recurrent spontaneous abortion.

BACKGROUND: At present, it is generally believed that immune factors account for 60% of unexplained recurrent spontaneous abortion (URSA). The treatments used for URSA depend on immunomodulation for their effects, and paternal immunization, intravenous immunoglobulin, and the use of growth factors such as granulocyte-colony stimulating factor (filgrastim) have been shown to have a beneficial effect on patients with a poor prognosis. However, these treatment schemes and effects remain controversial. This study aimed to evaluate the effect of immunotherapy using lymphocyte active immunotherapy (LAI) on patients with URSA, and to provide evidences for the clinical effect of this treatment. METHODS: The detailed data of total 619 patients with URSA were collected and analyzed, of which 465 patients (LAI group) with immunotherapy and 154 patients (control group) without immunotherapy. RESULTS: After 77.6% of all the patients in LAI group received the immunotherapy, the maternal blocking antibody (BA) was changed from negative to positive. The conversion rate of maternal BA was increased as the increase of active immunization (>4 times, P<0.05). The pregnancy rate of LAI Group was higher than that of the control group (P<0.05), and there were significant differences of live rate and abortion rate (P<0.05). In addition, compared with the natural pregnancy, the live rate was higher, and the abortion rate was lower in in vitro fertilization (IVF) patients after active immunization, although the difference was not significant (P>0.05). CONCLUSIONS: After lymphocyte immunotherapy, most of the patients with unexplained recurrent spontaneous abortion had the positive BA instead of negative BA. Whether the BA was converted or not, the pregnancy rate and live rate were increased, and the abortion rate was decreased after immunotherapy. Therefore, active immunotherapy could improve the pregnancy outcome of the patients with unexplained recurrent abortion.

[Expression of estrogen receptor, progesterone receptor and integrin alphaVbeta3 in endometrium of patients with luteinized unruptured follicle.].

OBJECTIVE: To investigate endometrium receptivity in patients with luteinized unruptured follicle (LUF) by measuring the expression of estrogen receptor (ER), progesterone receptor (PR) and integrin alphaVbeta3 in the endometrium. METHODS: From May 2007 to Nov. 2007, 17 infertile women with LUF were selected as LUF group matched with 13 infertile cases with normal ovulation as control group. They all underwent frozen-thawed embryo transfer in Reproductive Medicine Center, Renmin Hospital of Wuhan University. Endometrial tissue in anterior and posterior wall of uterus of LUF group and control group were biopsied by a small curettage between 7 and 11 days after luteinizing hormone (LH) surge. The expression of ER, PR and integrin alphaVbeta3 in endometrium were detected by immunohistochemistry staining. The level of estrogen and progesterone were measured by chemiluminescence assay. Then, the relationship between alphaVbeta3 expression in endometrium and the level of estrogen/progesterone were analyzed in LUF patients. RESULTS: (1) There was no remarkable difference in the level of estrogen between LUF [(656 +/- 299) pmol/L] and control group [(727 +/- 275) pmol/L, P > 0.05]. However, the level of progesterone were (23 +/- 8) nmol/L in LUF group and (35 +/- 10) nmol/L in control group, which reached statistical difference (P < 0.01). (2) The expression of ER, PR in endometrium of LUF patients were 183.9 +/- 2.4 and 168 +/- 3, which were significantly higher than 109.4 +/- 6.3 and 106 +/- 4 in control group (P < 0.01). The expression of integrin alphaVbeta3 in endometrium of 115 +/- 11 in LUF group were significantly lower than 191 +/- 9 in control group (P < 0.01). (4) In LUF group, the expression of alphaVbeta3 in endometrium was correlated positively with the level of progesterone (r = 0.77, P < 0.01) and irrelevant with the level of estrogen (r = 0.01, P > 0.05). CONCLUSION: The higher expression of estrogen and progesterone and lower expression of integrin alphaVbeta3 might confer impaired receptivity of endometrium and interfere with embryo implantation.

Long non-coding RNAs in diseases related to inflammation and immunity.

Long non-coding RNAs (lncRNAs) have been key regulators of gene expression in innate and adaptive immunity. Although lncRNAs have been reported to be associated with some diseases, its expression and function in diseases related to inflammation and immunity are still unknown. We reviewed how lncRNA regulated transcription and controlled the function and balance of the cells in the immune response. In addition, we discussed the impacts and challenges of lncRNAs on immunity in diseases.

The Increased lncRNA MIR503HG in Preeclampsia Modulated Trophoblast Cell Proliferation, Invasion, and Migration via Regulating Matrix Metalloproteinases and NF-κB Signaling.

BACKGROUND: Preeclampsia (PE) is a pregnancy-related syndrome characterized by hypertension and proteinuria after the 20th week of gestation. The long noncoding RNAs (lncRNAs) have been recently discovered for their roles in the pathogenesis of PE. This study is aimed at determining the expression of lncRNA MIR503 host gene (MIR503HG) in PE placental tissues and exploring the molecular mechanism underlying MIR503HG-mediated trophoblast cell proliferation, invasion, and migration. METHODS: The expression level of MIR503HG in placental tissues, HTR-8/SVneo, and JEG3 cells was determined by quantitative real-time PCR; western blot detected the relevant protein expression levels in HTR-8/SVneo and JEG3 cells; flow cytometry determined cell apoptosis and cell cycle of HTR-8/SVneo and JEG3 cells; trophoblast cell proliferation, invasion, and migration of HTR-8/SVneo and JEG3 cells were measured by CCK-8, transwell invasion, and wound healing assays, respectively. RESULTS: The highly expressed MIR503HG was detected in PE placental tissues compared to normal placental tissues. MIR503HG overexpression suppressed cell proliferation, invasion, and migration of HTR-8/SVneo and JEG3 cells, while knockdown of MIR503HG increased trophoblast cell proliferation, invasion, and migration. Flow cytometry results showed that MIR503HG overexpression induced apoptosis and caused cell cycle arrest at the G0/G1 phase, while MIR503HG knockdown had the opposite actions in HTR-8/SVneo and JEG3 cells. Western blot assay results showed that MIR503HG overexpression suppressed the matrix metalloproteinase-2/-9 and the snail protein expression and increased the E-cadherin expression in trophoblast cells. In addition, MIR503HG overexpression suppressed the NF-κB signaling pathway by inhibiting the phosphorylation of IκBα and the nuclear translocation of NF-κB signaling subunit p65. On the other hand, MIR503HG knockdown played an opposite role in these protein expression levels. CONCLUSION: Our results showed that MIR503HG inhibited the proliferation, invasion, and migration of HTR-8/SVneo and JEG3 cells, which may be related to the pathogenesis of PE.

The high-risk HPV oncogene E7 upregulates miR-182 expression through the TGF-ß/Smad pathway in cervical cancer.

Accumulating experimental evidence has shown that the aberrant expression of microRNAs (miRNAs) is involved in the development and progression of human cervical cancer. Previously, we identified miR-182 as an oncomiRNA in cervical cancer. However, the mechanism by which miR-182 is regulated and the interaction between human papillomavirus (HPV) and miR-182 in cervical cancer development remains unknown. In the present study, we explored the link between HPV E7 and miR-182 and verified that high-risk HPV E7 upregulated miR-182 expression through TGF-ß/Smad4 signaling pathway in cervical cancer. By contrast, low-risk HPV E7 did not affect the expression of TGF-ß and miR-182. Mechanistically, as high-risk HPV E7 bound to pRb, E2F was released from the complex and bound to the TGF-ß promoter region, resulting in TGF-ß overexpression. Furthermore, the Smad4 signaling pathway was activated upon TGF-ß overexpression, which led to an interaction between Smad4 and the miR-182 promoter region, subsequently inducing the upregulation of miR-182 in both cervical cancer cells and the surrounding normal cells. In conclusion, this newly identified high-risk HPV E7/TGF-ß/miR-182 regulatory network might inform the development of specific therapeutic strategies for cervical cancer.