3D printing technique applied in vaginoplasty: A case report.

Background: We present the first case report of the treatment of congenital vaginal atresia by 3D-printed patient-specific vaginal scaffold from China. Case presentation: A 17-year-old female patient was referred to our department for treatment of congenital vaginal atresia and complications arising from previous failed operations. Pelvic examination was conducted to understand the morphological characteristics and severity of stenosis, and based on which we designed our prototypes of vaginal scaffold using software UG NX10.0. We finally obtained our patient-specific mold, which was 50 mm in length, 28 mm in diameter, 2 mm of thickness with a whole weight of 7.6 g, and it was made of polycaprolactone. After removing scar tissues caused by vaginal stenosis, an 8 cm long artificial tunnel was created, and then the polycaprolactone (PCL) vaginal mold was placed and sutured. The patient had no discomfort after surgery and was discharged 3 days after the surgery. Follow-up for 1 year after surgery, through hysteroscopy and colposcopy, it was found that the cervix was smooth, the vaginal wall was covered with stratified squamous epithelium, and the vaginal wall was soft and lubricated, which was close to a normal vagina. The incompletely absorbed mold was taken out one year after the operation. Hysteroscopy and colposcopy were performed one year and two years after the mold was taken out. The vagina was unobstructed and the length was about 12 cm. The appearance of the vaginal wrinkles was normal. The patient's quality of sexual life was good. Conclusion: Our team tried to treat congenital vaginal atresia by 3D-printed patient-specific vaginal scaffold, which can effectively reduce patient complications and reduce patient pain. Through long-term follow-up, we found that this technique has achieved favorable results and improved the patient's quality of sexual life.

Suppression of microRNA-18a expression inhibits invasion and promotes apoptosis of human trophoblast cells by targeting the estrogen receptor α gene.

The purpose of the present study was to gain further understanding of the function of microRNA-18a (miR-18a) expression in the JEG-3 human trophoblast cell line. JEG-3 cells were transfected with pre-miR-18a mimics or miR-18a inhibitors. The effects of miR-18a on trophoblast cell invasion and apoptosis, and on the expression of estrogen receptor α (ESRα) were analyzed using a Transwell invasion assay, flow cytometry, reverse transcription-polymerase chain reaction, western blot analysis and a luciferase assay. The results of the present study suggested that miR-18a expression suppression led to a decrease in JEG-3 cell invasion and an increase in JEG-3 cell apoptosis, by inducing ESRα expression. The present study provides evidence for the involvement of miR-18a in the pathogenesis of pre-eclamptic pregnancies.

Role of HLA-G1 in trophoblast cell proliferation, adhesion and invasion.

Trophoblast cells are important in embryo implantation and fetomaternal tolerance. HLA-G is specifically expressed at the maternal-fetal interface and is a regulator in pregnancy. The aim of the present study was to detect the effect of HLA-G1 on trophoblast cell proliferation, adhesion, and invasion. Human trophoblast cell lines (JAR and HTR-8/SVneo cells) were infected with HLA-G1-expressing lentivirus. After infection, HLA-G1 expression of the cells was detected by western blotting. Cell proliferation was detected by the BrdU assay. The cell cycle and apoptosis of JAR and HTR-8/SVneo cells was measured by flow cytometry (FCM). The invasion of the cells under different conditions was detected by the transwell invasion chamber assay. HLA-G1 didn't show any significant influence on the proliferation, apoptosis, adhesion, and invasion of trophocytes in normal culture conditions. However, HLA-G1 inhibited JAR and HTR-8/SVneo cells invasion induced by hepatocyte growth factor (HGF) under normal oxygen conditions. In conditions of hypoxia, HLA-G1 couldn't inhibit the induction of cell invasion by HGF. HLA-G1 is not an independent factor for regulating the trophocytes. It may play an indirect role in embryo implantation and formation of the placenta.

Differential expression of estradiol and estrogen receptor α in severe preeclamptic pregnancies compared with normal pregnancies.

The purpose of this study was to investigate the expression of estradiol and estrogen receptor α (ESRα) in severe preeclamptic (sPE) pregnancies compared with normal pregnancies. Sera and placentas were obtained from i) patients with sPE (n=25) and ii) a normal control group (n=25) who underwent elective Cesarean deliveries. Estradiol expression was assessed by enzyme-linked immunosorbent assays (ELISAs). ESRα expression was assessed by reverse transcription polymerase chain reaction (RT-PCR) analysis and western blot analysis. In preeclamptic pregnancies, estradiol was underexpressed (P<0.05), however, ESRα mRNA and protein levels were increased significantly in comparison with normal pregnancies (P<0.05). These results show that estradiol and ESRα are deregulated in preeclamptic pregnancies, which in turn suggests the involvement of these molecules in the pathogenesis of preeclampsia.

Matrix metalloproteinase-9 is a prognostic marker for patients with cervical cancer.

Cervical cancer remains one of the most common malignancies in women. Previous study proved MMP-9 might be prognostic marker for multiple human malignancies. The present study was to investigate the protein expression of MMP-9 in cervical cancer and its association with clinicopathological characteristics as well as prognosis of patients. Cervical cancer specimens from 225 cases who had not received chemotherapy or radiotherapy prior to surgery were collected. Immunochemistry assays were utilized to investigate MMP-9 protein expression. Results showed that MMP-9 expression was increased in cervical cancer and associated with stromal invasion, FIGO stage, lymph node metastasis, and vascular invasion. Kaplan-Meier analysis showed that patients with cervical cancer of positive MMP-9 staining tend to have worse overall survival. In multivariate analysis stratified for known prognostic variables, MMP-9 was proved to be an independent prognostic factor. The present study confirmed that MMP-9 expression in cervical cancer was an independent prognostic factor of patients, which might be a potential diagnostic and even therapeutic target of cervical cancer.

Expression of human leukocyte antigen-G during normal placentation and in preeclamptic pregnancies.

OBJECTIVES: This study was undertaken to investigate the expression pattern of human leukocyte antigen-G (HLA-G) during normal placentation and determine whether altered expression of HLA-G is associated with severe preeclampsia. METHODS: We investigated HLA-G protein levels in first (n = 27), second (n = 7), and third trimester placentas (n = 10) from normal pregnancies, and determined HLA-G levels in term placentas from normal (n = 15) and severe preeclamptic pregnancies (n = 14) using real-time RT-PCR and western blot analysis. RESULTS: In normal placentas, HLA-G protein expression reached a peak level at gestational weeks 6 and 7, then gradually decreased from week 8 to third trimester (p < 0.05). In preeclamptic placentas, both HLA-G mRNA and protein levels were decreased significantly in comparison with normal term placentas (p < 0.05). CONCLUSION: HLA-G may contribute to placentation during early and mid-term pregnancy, and participate in maintaining gestation during term pregnancy. The reduced level of HLA-G may be associated with pathogenesis of preeclampsia.

Rapid and efficient reprogramming of human amnion-derived cells into pluripotency by three factors OCT4/SOX2/NANOG.

Reprogramming human somatic cells to pluripotency represents a valuable resource for research aiming at the development of in vitro models for human diseases and regenerative medicines to produce patient-specific induced pluripotent stem (iPS) cells. Seeking appropriate cell resources for higher efficiency and reducing the risk of viral transgene activation, especially oncogene activation, are of significance for iPS cell research. In this study, we tested whether human amnion-derived cells (hADCs) could be rapidly and efficiently reprogrammed into iPS cells by the defined factors: OCT4/SOX2/NANOG. hADCs from normal placenta were isolated and cultured. The 3rd passage cells were infected with the lentiviral vectors for the delivery of OCT4, SOX2, and NANOG. Afterwards, the generated iPSCs were identified by morphology, pluripotency markers, global gene expression profiles, and epigenetic status both in vitro and in vivo. The results showed that we were able to reprogram hADCs by the defined factors (OCT4/SOX2/NANOG). The efficiency was significantly high (about 0.1%), and the typical colonies appeared on the 9th day after infection. They were similar to human embryonic stem (ES) cells in morphology, proliferation, surface markers, gene expression, and the epigenetic status of pluripotent cell-specific genes. Furthermore, these cells were able to differentiate into various cell types of all three germ layers both in vitro and in vivo. These results demonstrate that hADCs were an ideal somatic cell resource for the rapid and efficient generation of iPS cells by OCT4/SOX2/NANOG.

Overexpression of miR-152 leads to reduced expression of human leukocyte antigen-G and increased natural killer cell mediated cytolysis in JEG-3 cells.

OBJECTIVE: The purpose of this study was to gain a further understanding of the relationship between miR-152 and human leukocyte antigen (HLA)-G in human trophoblast cell line (JEG-3). STUDY DESIGN: The JEG-3 cells were transfected with pre-miR-152. The effect of the overexpressed miR-152 on HLA-G expression, trophoblast invasion, and natural killer (NK) cell-mediated cytolysis were assessed by reverse-transcription polymerase chain reaction (RT-PCR) and Western blot analysis, transwell invasion assay, and NK cell cytotoxicity assay, respectively. RESULTS: The miR-152 repressed HLA-G expression but exerted no effect on JEG-3 cell invasion, and overexpression of miR-152 led to increased NK cell-mediated cytolysis in JEG-3 cells. CONCLUSION: The data indicate that miR-152 may function as an immune system enhancer through up-regulating NK cell-mediated cytolysis of host cells.

Differential expression profile of microRNAs in human placentas from preeclamptic pregnancies vs normal pregnancies.

OBJECTIVE: The purpose of this study was to perform a comprehensive analysis of the microRNA expression profile in placentas from preeclamptic pregnancies vs normal placentas. STUDY DESIGN: Placentas were obtained from patients with (1) mild preeclampsia (n = 8) and (2) severe preeclampsia (n = 15) and (3) in a normal control group (n = 11) with elective cesarean delivery. The microRNA expression profile was assessed by microRNA microarray and real-time reverse transcriptase-polymerase chain reaction analysis. RESULTS: Thirty-four microRNAs were expressed differentially in preeclamptic placentas, compared with normal placentas. Of these, 11 microRNAs were overexpressed, and 23 microRNAs were underexpressed in preeclamptic pregnancies. Notably, several microRNA clusters on human chromosome 19q13.42, 13q31.3, Xq26.2, Xq26.3, and 14q32.31 (a human imprinted region) were expressed differentially in preeclamptic placentas. These results were confirmed with the use of real-time polymerase chain reaction for selected microRNAs (miR-210, -152, -411, and so on). CONCLUSION: The results show that 34 microRNAs are deregulated in preeclamptic pregnancies, which suggests the involvement of these microRNAs in the pathogenesis of preeclampsia.

Determination of microRNAs in mouse preimplantation embryos by microarray.

Expression profile of microRNA (miRNA) in mouse oocytes and preimplantation embryos has been revealed by a novel high throughput microarray assay. A total of 97 (43 "new" and 54 known) including mouse, human, and predicted miRNAs have been discovered in the preimplantation mouse embryos which can be classified into developmental stage-dependent groups and non-stage-dependent group according to the statistical analysis of the expression patterns. Potential gene targets of each group of miRNAs are estimated by TargetsScan system and significantly changed signaling pathways and biological processes underlying these gene targets are searched by PANTHER classification system between the stage-dependent miRNAs and the non-stage-dependent miRNAs. Expression of some miRNAs is confirmed by reverse transcriptase-polymerase chain reaction. It is shown that dynamic synthesis and degradation of miRNAs coexists in the preimplantation development of mouse embryos. However, the overall quantity of miRNAs and percentage of the stage-dependent miRNAs increase as the preimplantation embryos develop.

[Expression of hypoxia inducible factor-1alpha in placenta of pregnancy induced hypertension].

OBJECTIVE: To investigate the expression of hypoxia inducible factor-1alpha (HIF-1alpha) in placentas of pregnancy induced hypertension (PIH). METHODS: Placentas of 49 PIH pregnancies (PIH group 1) and 26 normotensive pregnancies (control group 1) were investigated for HIF-1alpha protein expression using microarray and immunohistochemistry. The expression of HIF-1alpha mRNA in placentas of 12 PIH pregnancies (PIH group 2) and 12 normal pregnancies(control group 2) was respectively detected by RT-PCR. RESULTS: (1) There was significant difference in the positive and slight-moderate-positive spot proportions of HIF-1alpha protein between the PIH group 1 (63.1%, 23.3%) and the control group 1 (20.9%, 7.0%, P < 0.01). (2) The level of HIF-1alpha mRNA between PIH group 2 and control group 2 (0.96 +/- 0.37 vs 0.95 +/- 0.20, P > 0.05) was not statistically different. CONCLUSIONS: (1) There was no significant difference in HIF-1alpha mRNA level between the PIH and normal placentas, but there was a remarkable difference in protein level between the two groups. (2) HIF-1alpha regulate the pathological and physiological progress of PIH at protein level rather than at transcription level.

Successful rescuing a pregnant woman with severe hepatitis E infection and postpartum massive hemorrhage.

AIM: To sum up the experience of the successful therapy for the severe hepatitis of pregnant woman with postpartum massive hemorrhage. METHODS: The advanced therapeutic methods including the bilateral uterine artery embolism, hemodialysis and artificial liver support therapy were performed with comprehensive medical treatments and the course of the successful rescuing the patient was analyzed. RESULTS: Through the hospitalization of about two mouths the patient and her neonatus had gotten the best of care in our department and pediatric department separately. Both of them were discharged in good condition. CONCLUSION: The key points for a successful therapy of the pregnant woman with severe hepatitis are termination of the pregnancy and the control of their various complications. It was suggested that the proper combination of these measures of modern therapy would race against time for renewing of hepatic and renal functions.