Foxf2 and Smad6 co-regulation of collagen 5A2 transcription is involved in the pathogenesis of intrauterine adhesion.

The replacement of normal endometrial epithelium by fibrotic tissue is the pathological feature of intrauterine adhesion (IUA), which is caused by trauma to the basal layer of the endometrium. COL5A2 is a molecular subtype of collagen V that regulates collagen production in fibrotic tissue. Here, we investigated the roles of Foxf2 and Smad6 in regulating the transcription of COL5A2 and their involvement in the pathogenesis of IUA. Small interference-mediated Foxf2 (si-Foxf2) silencing and pcDNA3.1-mediated Smad6 (pcDNA3.1-Smad6) up-regulation were performed in a TGF-ß1-induced human endometrial stromal cell line (HESC) fibrosis model. Assessment of collagen expression by Western blotting, immunofluorescence and qRT-PCR showed that COL5A2, COL1A1 and FN were significantly down-regulated in response to si-Foxf2 and pcDNA3.1-Smad6. Transfection of lentivirus vector-Foxf2 (LV-Foxf2) and pcDNA3.1-Smad6 into HESCs and qRT-PCR showed that Foxf2 promoted COL5A2 expression and Smad6 inhibited Foxf2-induced COL5A2 expression. Co-immunoprecipitation, chromatin immunoprecipitation and dual-luciferase reporter assays to detect the interaction between Foxf2 and Smad6 and their role in COL5A2 transcription showed that Foxf2 interacted with Smad6 and bond the same promoter region of COL5A2. In a rat IUA model, injection of ADV2-Foxf2-1810 and ADV4-Smad6 into the uterine wall showed that Foxf2 down-regulation and Smad6 up-regulation decreased fibrosis and the expression of COL5A2 and COL1A1, as detected by haematoxylin/eosin, Masson trichrome staining and immunohistochemistry. Taken together, these results suggested that Foxf2 interacted with Smad6 and co-regulated COL5A2 transcription in the pathogenesis of IUA, whereas they played opposite roles in fibrosis.

Exosomal transfer of miR-124 inhibits normal fibroblasts to cancer-associated fibroblasts transition by targeting sphingosine kinase 1 in ovarian cancer.

OBJECTIVE: The interaction between tumor microenvironment and tumor cells plays a key role in tumor progression. However, the mechanisms by which this interaction promotes the transdifferentiation of normal fibroblasts (NFs) to cancer-associated fibroblasts (CAFs) are still unclear. The aim of this study was to investigate whether ovarian cancer (OvCa) cells-derived microRNAs were involved in the transition of resident fibroblasts to CAFs, and in promoting tumorigenesis. METHODS: CAFs and NFs were isolated from the same ovarian site in OvCa and noncancerous prophylactic oophorectomy specimens. The effect of exosomes on the motility of CAFs or NFs was analyzed by wound healing and Transwell assays. The expression of CAFs marker α-smooth muscle actin (α-SMA) and fibroblast activated protein (FAP) were determined by quantitative real-time PCR and Western blotting. A luciferase reporter assay was used to test the interaction between miR-124 and sphingosine kinase 1 (SPHK1). RESULTS: NFs with downregulated miR-124 displayed the characteristics of CAFs, including overexpression of α-SMA and FAP and increased migratory and invasive ability. Overexpression of miR-124 in CAFs reversed some traits of NFs. Human ovarian surface epithelial cells-secreted miR-124 could be transferred via exosomes to CAFs and resulted in decreased α-SMA and FAP expression and attenuated cell motility. Moreover, our finding showed that the expression of SPHK1, a potential target of miR-124, was significantly elevated in CAFs. CONCLUSIONS: The present study provides important and novel perspective into OvCa CAF differentiation and extracellular matrix remodeling, which trigger the tumor progression.

MicroRNA-16 Promotes Ovarian Granulosa Cell Proliferation and Suppresses Apoptosis Through Targeting PDCD4 in Polycystic Ovarian Syndrome.

BACKGROUND/AIMS: Several miRNAs have been reported to be involved in the pathogenesis of polycystic ovarian syndrome (PCOS). However, the biological roles of miR-16 and its molecular mechanisms in PCOS development remain to be elucidated. METHODS: qRT-PCR was performed to detect the expression levels of miR-16 and programmed cell death protein 4 (PDCD4). GCs proliferation, cell cycle distribution and apoptosis were examined by MTT assay and flow cytometry analysis. Luciferase reporter assay and RIP assay were applied to confirm the regulatory relationship between miR-16 and PDCD4. Western blot was applied to measure the protein levels of PDCD4, PCNA and caspase-3. ELISA kits were used to determine the serum levels of steroids. RESULTS: miR-16 expression was down-regulated in ovarian cortex tissues and serums of PCOS patients. PDCD4 expression was up-regulated in ovarian cortex tissues of PCOS patients. miR-16 overexpression facilitated cell proliferation, induced cell cycle progression, and inhibited apoptosis in GCs. Moreover, PDCD4 was a direct target of miR-16. Also, enforced expression of PDCD4 abated the effects of miR-16 on GCs growth and apoptosis. Additionally, testosterone resulted in a decrease of miR-16 expression and an increase of PDCD4 expression, thus blocking cell growth and enhanced apoptosis in GCs. Furthermore, miR-16 overexpression alleviated PCOS in vivo by regulating PDCD4. CONCLUSIONS: miR-16 promoted ovarian GCs proliferation and inhibited apoptosis through directly targeting PDCD4 in PCOS, contributing to a better understanding of the molecular mechanism of GCs dysregulation and providing a promising target in PCOS.

Oxidized, Regenerated Cellulose Adhesion Barrier Plus Intrauterine Device Prevents Recurrence After Adhesiolysis for Moderate to Severe Intrauterine Adhesions.

STUDY OBJECTIVE: To compare the efficacy of an oxidized, regenerated cellulose adhesion barrier (Interceed; Ethicon, Somerville, NJ) combined with an intrauterine device (IUD) versus an IUD alone for preventing adhesion recurrence following hysteroscopic adhesiolysis for moderate to severe intrauterine adhesions (IUAs). DESIGN: Retrospective case series (Canadian Task Force classification III). SETTING: Tertiary care teaching hospital. PATIENTS: Patients undergoing treatment for moderate to severe IUAs. The severity of IUA was determined based on the American Fertility Society scoring system (mild, moderate, or severe). INTERVENTIONS: All cases of hysteroscopic adhesiolysis were reviewed. MEASUREMENTS AND RESULTS: Seventy-six women with moderate to severe IUAs treated between March 2009 and August 2015 were included. After hysteroscopic adhesiolysis, 35 patients were treated with an IUD alone (group 1), and 41 patients were treated with Interceed plus an IUD (group 2). A second hysteroscopy was performed in all cases three months after the initial hysteroscopy and both groups achieved significant reduction in adhesion scores and grade, especially in group 2 (scores, p < .001; grade, p = .039). Compared with group 1, menstruation dysfunction, pregnancy rate, and live birth rate in group 2 improved with no statistical difference (menstruation improvement, p = .764; pregnancy rate, p = .310; live birth rate, p = .068). However, an adhesion-free uterine cavity was regained significantly owing to the fewer operations in group 2 compared with group 1 (median, 3 vs 4; p = .001). The interval from initial hysteroscopy to conception was significantly shorter in group 2 (median, 12 months vs 51 months; p < .001). CONCLUSIONS: For moderate to severe IUAs, Interceed combined with an IUD may be an alternative approach for reducing adhesion recurrence after hysteroscopic adhesiolysis.

MicroRNA-155 promotes apoptosis in SKOV3, A2780, and primary cultured ovarian cancer cells.

MicroRNAs (miRNAs) are a large group of small non-coding RNAs that can negatively regulate gene expression at the post-transcriptional level. The deregulation of miRNAs has been associated with tumorigenesis, drug resistance, and prognosis in cancers. Deregulated miR-155 has been reported in numerous cancers; however, its function remains unclear. 4',6-Diamidino-2-phenylindole (DAPI) staining and terminal-deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) techniques were used to determine the effects of a miR-155 mimic or inhibitor on the apoptotic ratio of ovarian cancer cells induced by cisplatin. Bioinformatic predictions, the dual-luciferase reporter assay, and western blot analysis were used to detect how miR-155 regulates X-linked inhibitor of apoptosis protein (XIAP). We demonstrated that a miR-155 mimic could decrease the IC50 value of cisplatin in SKOV3 ovarian cancer cells. Subsequently, gain- and loss-of-function analyses with a miR-155 mimic and inhibitor showed that miR-155 sensitizes ovarian cancer cells to cisplatin. Furthermore, the results from the luciferase assays and western blot analysis identified XIAP as the direct target of miR-155. In addition, introducing XIAP cDNA without a three prime untranslated region (3'-UTR) rescued the miR-155 promotion of apoptosis. These results indicate that miR-155 mediates cisplatin-induced apoptosis by targeting XIAP in ovarian cancer cells and that miR-155 could be a potential therapeutic target to increase the efficiency of ovarian cancer interventions.

[Resource investigation of wild Codonopsis pilosula in Tanchang county of Gansu].

Tanchang county is the distribution of wild medicinal plant resource-rich region, in order to ascertain Tanchang county Codonopsis pilosula wild resources and reserves of the status quo, according to the fourth national Chinese medicine resources survey technology solutions, using sets of plots and investigating combined route survey method, the county wild C. pilosula var. modesta and C. pilosula resources were investigated by a comprehensive survey designed to reveal the distribution of the county's wildlife resources and herbs C. pilosula reserves. The results showed that in Tanchang county seven ecological zones 53 plots, wild C. pilosula distributed in there were 6 ecological zones 11 plots, accounting for 85.71% of the survey area, wild C. pilosula var. modesta was found only in an ecoregions plots, overlapping with C. pilosula region, accounting for 14.29% of the survey area. C. pilosula herbs reserves were calculated as about 461.85 t, economic capacity of 254.02 t, annual amount of acceptance 25.40 t. C. pilosula var. modesta herbs reserves were calculated as 67.75 t, economic capacity of 36.16 t, acceptance annual amount 3.62 t. The total ash C. pilosula was 3.25%, alcohol-soluble extract was 63.86%, while the C. pilosula var. modesta total ash was 3.69%, alcohol-soluble extract was 68.32%. C. pilosula is suitable for broad range, but wild resource is scarce, C. pilosula var. modesta is suitable for relatively narrow scope, and wild resource is scarce, it is recommended to strengthen the protection of wild resources and the rational development and utilization.

Human umbilical cord mesenchymal stem cell transplantation restores damaged ovaries.

Ovarian injury because of chemotherapy can decrease the levels of sexual hormones and potentia generandi of patients, thereby greatly reducing quality of life. The goal of this study was to investigate which transplantation method for human umbilical cord mesenchymal stem cells (HUMSCs) can recover ovarian function that has been damaged by chemotherapy. A rat model of ovarian injury was established using an intraperitoneal injection of cyclophosphamide. Membrane-labelled HUMSCs were subsequently injected directly into ovary tissue or tail vein. The distribution of fluorescently labelled HUMSCs, estrous cycle, sexual hormone levels, and potentia generandi of treated and control rats were then examined. HUMSCs injected into the ovary only distributed to the ovary and uterus, while HUMSCs injected via tail vein were detected in the ovary, uterus, kidney, liver and lung. The estrous cycle, levels of sex hormones and potentia generandi of the treated rats were also recovered to a certain degree. Moreover, in some transplanted rats, fertility was restored and their offspring developed normally. While ovary injection could recover ovarian function faster, both methods produced similar results in the later stages of observation. Therefore, our results suggest that transplantation of HUMSCs by tail vein injection represents a minimally invasive and effective treatment method for ovarian injury.

Comparison of laparoscopy and laparotomy in the surgical management of early-stage ovarian cancer.

OBJECTIVE: The aim of this study is to investigate the safety and efficacy of laparoscopy in the treatment of early-stage ovarian cancer (EOC). METHODS: We retrospectively analyzed the clinical data of patients who underwent laparoscopy (35 patients) or laparotomy (40 patients) for the comprehensive surgical staging of EOC in Zhujiang Hospital during the period of 2002 to 2010 and compared the 2 surgical approaches in operative time, intraoperative blood loss, number of dissected lymph nodes, tumor rupture rate, length of hospital stay, time of gastrointestinal function recovery, wound healing condition, complication rate, upstaging rate, rate of postoperative chemotherapy, and postoperative follow-up condition. RESULTS: The laparoscopy group had significantly shorter hospital stay and time of first postoperative flatus and had significantly lower rate of poor wound healing than the laparotomy group. The 2 groups did not show significant differences in operative time, intraoperative blood loss, number of dissected lymph nodes, tumor rupture rate, complication rate, upstaging rate, and rate of postoperative chemotherapy. CONCLUSIONS: Laparoscopy is safe and effective for the comprehensive surgical staging of EOC and has the advantages of shorter hospital stay, faster recovery of gastrointestinal function, and good wound healing.

IGF2BP3 enhances lipid metabolism in cervical cancer by upregulating the expression of SCD.

Cervical cancer (CC) is the most common gynecologic malignancy, which seriously threatens the health of women. Lipid metabolism is necessary for tumor proliferation and metastasis. However, the molecular mechanism of the relationship between CC and lipid metabolism remains poorly defined. We revealed the expression of IGF2BP3 in CC exceeded adjacent tissues, and was positively associated with tumor stage using human CC tissue microarrays. The Cell Counting Kit-8, colony formation assay, 5-ethynyl-2'-deoxyuridine assay, transwell assays, wound-healing assays, and flow cytometry assessed the role of IGF2BP3 in proliferation and metastasis of CC cells. Besides, exploring the molecular mechanism participating in IGF2BP3-driven lipid metabolism used RNA-seq, which determined SCD as the target of IGF2BP3. Further, lipid droplets, cellular triglyceride (TG) contents, and fatty acids were accessed to discover that IGF2BP3 can enhance lipid metabolism in CC. Moreover, RIP assay and methylated RNA immunoprecipitation experiments seeked the aimed-gene-binding specificity. Lastly, the IGF2BP3 knockdown restrained CC growth and lipid metabolism, after which SCD overexpression rescued the influence in vitro and in vivo using nude mouse tumor-bearing model. Mechanistically, IGF2BP3 regulated SCD mRNA m6A modifications via IGF2BP3-METTL14 complex, thereby enhanced CC proliferation, metastasis, and lipid metabolism. Our study highlights IGF2BP3 plays a crucial role in CC progression and represents a therapeutic latent strategy. It is a potential tactic that blocks the metabolic pathway relevant to IGF2BP3 with the purpose of treating CC.

Creation of a female rabbit model for intrauterine adhesions using mechanical and infectious injury.

BACKGROUND: Intrauterine adhesions (IUA) are associated with secondary amenorrhea, infertility, and recurrent pregnancy loss. An IUA animal model would contribute to research on the pathogenesis and pathologic changes of IUA and the exploration of new treatments. MATERIALS AND METHODS: Eighty female rabbits were randomly divided into five groups: mechanical injury (16), infectious injury (16), dual injury (16), experimental control (16), and normal (16). Three methods were applied to establish the model: uterine curettage, uterine cavity left alone, lipopolysaccharide surgical suture in place for 48 h, and suture retention for 48 h after curettage. A sterile surgical suture was left in the uterine cavity for 48 h in the experimental control group. Histologic changes were monitored at 0, 24, 48, and 72 h and 7, 14, and 28 d after operation. RESULTS: The experiments revealed that endometrium injured by simple curettage or infection could be repaired. Although endometrial regeneration was severely impaired by dual injury, the ratio of the area with endometrial stromal fibrosis to total endometrial area significantly increased (P < 0.01), and the number of endometrial glands was significantly reduced (P < 0.01). CONCLUSIONS: The method of dual injury can establish a stable rabbit IUA model.

Characterization of a subtilisin-like protease with apical localization from microsporidian Nosema bombycis.

The microsporidian Nosema bombycis is the pathogen causing pébrine leading to heavy economic loss in sericulture. Little is known of the proteases of microsporidia that are important for both parasite development and pathogenesis. Here we identified a subtilisin-like serine protease NbSLP1 which contains an inhibitor_I9 and a peptidase_S8 domain. Three dimensional modeling of the catalytic domain of the NbSLP1 exhibited a typical 3-layer sandwich structure with S1 pocket substituted by Y(359). Phylogenetic analysis confirms that subtilisin-like serine proteases of microsporidia fall into two clades: SLP1 and SLP2, suggesting the initial subtilisin gene duplication events preceded microsporidia speciation. Furthermore, transcripts of Nbslp1 were detected in the midgut of Bombyx mori infection by N. bombycis by RT-PCR. Antibodies against NbSLP1 recognized both the precursor and mature enzyme by 2D Western blotting. Besides, indirect immunofluorescence assay revealed that the NbSLP1 is mainly localized at the two poles of spore which make the spore look like "safety pins". Remarkably, the mature protease is only detected in the apical region of the spore after germination. These studies demonstrate that NbSLP1 is a conserved subtilisin protease in microsporidia and suggest that NbSLP1 play a significant role in polar tube extrusion process.

The traditional uses, phytochemistry, pharmacology and toxicology of Bergenia purparescens: A review comments and suggestions.

Bergenia purpurascens (B. purpurascens, Saxifragaceae) has been used to treat several diseases in different countries, such as lung diseases, stomach problems, rheumatic pains, boosting immunity etc. However, the information on phytochemistry, pharmacology and toxicology of this plant has rarely been comprehensively and critically reported. This paper aims to study and evaluate its therapeutic potential, including the traditional uses and all the latest information of phytochemistry, pharmacology and toxicology. The main components of this plant are phenols compounds and the characteristic substance is bergenin.The results about modern pharmacology have shown that its pharmacological effects include antibacterial, antiviral, cough relieving, anti-inflammatory and so on. In addition, it could inhibit diabetic neuropathy, restore insulin secretion, treat cancer, protect liver and prevent Alzheimer's disease (AD). Thus, its therapeutic fields may be cancer, diabetic and AD in the future. The information will help to further update and study pharmacologic effect and action mechanism of this herb, which is more widely, effectively, and safely used in clinic.

Overexpression of the ß subunit of human chorionic gonadotropin promotes the transformation of human ovarian epithelial cells and ovarian tumorigenesis.

Ovarian carcinoma is the most lethal gynecologic malignancy, however underlying molecular events remain elusive. Expression of human chorionic gonadotropin ß subunit (ß-hCG) is clinically significant for both trophoblastic and nontrophoblastic cancers; however, whether ß-hCG facilitates ovarian epithelial cell tumorigenic potential remains uncharacterized. Immortalized nontumorigenic ovarian epithelial T29 and T80 cells stably overexpressing ß-hCG were examined for alterations in cell cycle and apoptotic status by flow cytometry, expression of proteins regulating cell cycle and apoptosis by Western blot, proliferation status by MTT assay, anchorage-independent colony formation, and mouse tumor formation. Immunoreactivity for ß-hCG was evaluated using mouse xenografts and on human normal ovarian, fallopian tube, endometrium, and ovarian carcinoma tissues. T29 and T80 cells overexpressing ß-hCG demonstrated significantly increased proliferation, anchorage-independent colony formation, prosurvival Bcl-X(L) protein expression, G2-checkpoint progression, elevated cyclins E/D1 and Cdk 2/4/6, and decreased apoptosis. Collectively, these transformational alterations in phenotype facilitated increased xenograft tumorigenesis (P < 0.05). Furthermore, ß-hCG immunoreactivity was elevated in malignant ovarian tumors, compared with normal epithelial expression in ovaries, fallopian tube, and endometrium (P < 0.001). Our data indicate that elevated ß-hCG transforms ovarian surface epithelial cells, facilitating proliferation, cell cycle progression, and attenuated apoptosis to promote tumorigenesis. Our results further decipher the functional role and molecular mechanism of ß-hCG in ovarian carcinoma. ß-hCG may contribute to ovarian cancer etiology, which introduces a new therapeutic intervention target for ovarian cancer.

[Biological features and ultrastructure of human umbilical cord mesenchymal stem cells].

OBJECTIVE: To isolate and culture human umbilical cord mesenchymal stem cells (MSCs) and explore their biological features and ultrastructure. METHODS: After isolating MSCs from the human umbilical cord, the proliferation, cycle, and apoptosis were observed. The cell ultrastructure was observed under transmission electron microscope. The cytokines including vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), and insulin-like growth factor-1 (IGF-1) were detected using enzyme-linked immunosorbent assay. RESULTS: Human umbilical cord MSCs had fibroblast-like morphology and increased proliferation capability. Ultrastructural analysis showed that the MSCs had active cellular metabolism and strong migration and differentiation capabilities. Meanwhile, they could secrete anti-apoptotic cytokines such as VEGF, IGF-1, and HGF. CONCLUSION: Human umbilical cord MSCs can secrete many anti-apoptotic cytokine and have good biological features.

Lactoferrin/Calcium Phosphate-Modified Porous Ti by Biomimetic Mineralization: Effective Infection Prevention and Excellent Osteoinduction.

The surface modification of titanium (Ti) can enhance the osseointegration and antibacterial properties of implants. In this study, we modified porous Ti discs with calcium phosphate (CaP) and different concentrations of Lactoferrin (LF) by biomimetic mineralization and examined their antibacterial effects and osteogenic bioactivity. Firstly, scanning electron microscopy (SEM), the fluorescent tracing method, X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR), energy dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), and the releasing kinetics of LF were utilized to characterize the modified Ti surface. Then, the antibacterial properties against S. sanguis and S. aureus were investigated. Finally, in vitro cytological examination was performed, including evaluations of cell adhesion, cell differentiation, extracellular matrix mineralization, and cytotoxicity. The results showed that the porous Ti discs were successfully modified with CaP and LF, and that the LF-M group (200 µg/mL LF in simulated body fluid) could mildly release LF under control. Further, the LF-M group could effectively inhibit the adhesion and proliferation of S. sanguis and S. aureus and enhance the osteogenic differentiation in vitro with a good biocompatibility. Consequently, LF-M-modified Ti may have potential applications in the field of dental implants to promote osseointegration and prevent the occurrence of peri-implantitis.

Analysis of endometrial microbiota in intrauterine adhesion by high-throughput sequencing.

BACKGROUND: Intrauterine adhesions (IUA) arise from scar tissue formation between the endometrial surfaces in response to mechanical or infectious injuries. However, the potential role of endometrial microbiota in IUA remains unclear. We aimed to explore the composition of endometrial microbiota and its potential role in IUA. METHODS: We retrospectively enrolled 46 patients diagnosed with IUA and 21 infertility patients without intrauterine lesions, as control subjects. All cases were diagnosed with hysteroscopy and endometrial tissues were taken from the intrauterine cavity using a hysteroscopic cutting ring without electricity study. After endometrial samples were collected, DNA was extracted and amplified for barcoded Illumina high-throughput next-generation sequencing targeted to the 16S rRNA V4 region for microbiota. Microbiota data were compared between two groups using α-diversity, ß-diversity and Nonmetric Multidimensional Scaling based on Weighted Unifrac distance. RESULTS: At the phyla level, the dominant bacteria included Proteobacteria, Firmicutes, Bacteroidetes and Actinobacteria. Proteobacteria accounted for more than 64.48%. At the genus level, the proportions of Klebsiella, Shewanella, and Lactobacillus were higher in patients with IUA than in non- IUA participants (20.67% and 8.77%, P=0.006, 13.37% and 4.53%, P=0.175, 12.74% and 6.95%, P=0.882; respectively). The proportion of Acinetobacter was significantly lower in patients with IUA than in non- IUA participants (P=0.005). CONCLUSIONS: Endometrial microbiota differ between patients with IUA and infertility patients without intrauterine lesions, and the potential variation of endometrial microbiota might cause IUA.

si-SNHG5-FOXF2 inhibits TGF-ß1-induced fibrosis in human primary endometrial stromal cells by the Wnt/ß-catenin signalling pathway.

BACKGROUND: Intrauterine adhesions (IUAs) are manifestations of endometrial fibrosis characterized by inflammation and fibrinogen aggregation in the extracellular matrix (ECM). The available therapeutic interventions for IUA are insufficiently effective in the clinical setting for postoperative adhesion recurrence and infertility problems. In this study, we investigated whether si-SNHG5-FOXF2 can serve as a molecular mechanism for the inhibition of IUA fibrosis ex vivo. METHODS: FOXF2, TGF-ß1 and collagen expression levels were measured by microarray sequencing analysis in three normal endometrium groups and six IUA patients. We induced primary human endometrial stromal cells (HESCs) into myofibroblasts (MFs) to develop an IUA cell model with various concentrations of TGF-ß1 at various times. Downstream target genes of FOXF2 were screened by chromatin immunoprecipitation combined with whole-genome high-throughput sequencing (ChIP-seq). We investigated ECM formation, cell proliferation and Wnt/ß-catenin signalling pathway-related proteins in primary HESCs with FOXF2 downregulation by quantitative reverse transcription-polymerase chain reaction (qRT-PCR), western blotting (WB), immunohistochemistry (IHC), flow cytometry, ethylenediurea (EdU) and CCK8 assays. We identified long noncoding RNAs (lncRNA) SNHG5 as the upstream regulatory gene of FOXF2 through RNA immunoprecipitation (RIP), RNA pulldown and fluorescence in situ hybridization (FISH). Finally, we examined FOXF2 expression, ECM formation, cell proliferation and Wnt/ß-catenin signalling pathway-related proteins in primary HESCs upon FOXF2 downregulation. RESULTS: FOXF2 was highly expressed in the endometrium of patients with IUA. Treatment of primary HESCs with 10 ng/ml TGF-ß1 for 72 h was found to be most effective for developing an IUA cell model. FOXF2 regulated multiple downstream target genes, including collagen, vimentin (VIM) and cyclin D2/DK4, by ChIP-seq and ChIP-PCR. FOXF2 downregulation inhibited TGF-ß1-mediated primary HESC fibrosis, including ECM formation, cell proliferation and Wnt/ß-catenin signalling pathway-related protein expression. We identified lncRNA SNHG5 as an upstream gene that directly regulates FOXF2 by RIP-seq, qRT-PCR, WB and FISH. SNHG5 downregulation suppressed FOXF2 expression in the IUA cell model, resulting in synergistic repression of the Wnt/ß-catenin pathway, thereby altering TGF-ß1-mediated ECM aggregation in endometrial stromal cells ex vivo. CONCLUSIONS: Regulation of the Wnt/ß-catenin signalling pathway and ECM formation by si-SNHG5-FOXF2 effectively inhibited the profibrotic effect of TGF-ß1 on primary HESCs. This finding can provide a molecular basis for antagonizing TGF-ß1-mediated fibrosis in primary HESCs.

Circular RNA hsa_circ_0003204 promotes cervical cancer cell proliferation, migration, and invasion by regulating MAPK pathway.

Cervical cancer (CC) is the second most common malignancy in women worldwide. The mechanism underlying CC development remains unclear. Recently, Circular RNAs (circRNAs)have attracted attention because of its role in tumorigenesis. To investigate circRNAsin CC, RNA sequencing was employed to characterize circRNA expression profile between CC tissues and matched adjacent normal tissues. The expression of hsa_circ_0003204 was examined in CC tissues and cell lines by real-time PCR. Migration assay and invasion assay were used to verify the effect of hsa_circ_0003204 on migration and invasion ability in CC cell lines. Tumor formation assay in nude mice was used to analyze the effect of hsa_circ_0003204 on the tumorigenicity of CC cell lines in vitro. Western blotting analyzes were performed to investigate the role of hsa_circ_0003204 in the regulation of MAPK signaling activation. We found that circRNA hsa_circ_0003204 was significantly upregulated in CC tissues. The function and potential molecular mechanisms of hsa_circ_0003204 were also investigated in vitro and in vivo. Hsa_circ_0003204 knockdown reduced cell growth, migration, and invasion but promoted cells apoptosis. However, the over-expression of hsa_circ_0003204 had the opposite effect. The MAPK pathway was different in hsa_circ_0003204 over-expression or down-expression cells, compared to parental cells. In addition, over-expression of hsa_circ_0003204 significantly increased tumor volume and tumor weight in vivo.Taken together, results indicated hsa_circ_0003204 may serve as a potential therapeutic target for patients with CC.

Aberrantly enhanced melanoma-associated antigen (MAGE)-A3 expression facilitates cervical cancer cell proliferation and metastasis via actuating Wnt signaling pathway.

BACKGROUND: The over-expression of melanoma-associated antigen (MAGE)-A3 in cervical cancer (CC) has been observed in our previous study, suggesting that it possibly take a vital role during the development and metastasis of CC. The present study aimed to investigate the biological function of MAGE-A3 in the progression of CC and explore how it executes its roles. METHODS: The mRNA expression of MAGE-A3 in End1/E6E7 and CC cell lines (HeLa, SiHa and C33A) was measured by real-time quantitative reverse transcription PCR (qRT-PCR). Loss- and gain-of-function methods were used to assess the effect of MAGE-A3 on the proliferative, migratory and invasive abilities of HeLa and SiHa cells. Western blot was performed to measure the expression levels of proteins related to epithelial-mesenchymal transition (EMT) and proteins in the Wnt signaling pathway. In vivo tumorigenesis assay was conducted to evaluate the effect of MAGE-A3 on tumor growth. RESULTS: MAGE-A3 expression was significantly up-regulated in CC cell lines (HeLa, SiHa and C33A) compared with that in End1/E6E7 cell line. Knockdown of MAGE-A3 could significantly suppress migration, invasion and proliferation in HeLa cells; whereas, overexpression of MAGE-A3 in SiHa cells presented the opposite results. Moreover, knockdown of MAGE-A3 presented a suppressive effect on the activation of EMT and Wnt signaling pathway in HeLa cells, whilst up-regulation of MAGE-A3 exhibited the opponent outcomes in SiHa cells. Through in vivo tumorigenesis assay, we further verified that MAGE-A3 acted as a facilitator in tumor growth. CONCLUSIONS: MAGE-A3 is overexpressed in CC cells and possibly facilitates the viability and motility of CC cells via modulating EMT and Wnt signaling. This study implied that MAGE-A3 might be a potential therapeutic target as well as a prognosis predictor for patients with CC.

[Analysis of Human Herpes Viruses-Activated Infection Spectra in Patients with Various Immunodeficiencies].

OBJECTIVE: To study the epidemiologic characteristics of human herpes virus (HHV) activated infection in the diseases of blood system and patients received allo-HSCT by statistically analyzing the screening results of 8 human herpes viruses (HHVs) of 4164 patients in Hebei Yanda LU Dao-Pei Hospital from 2012 to 2017. METHODS: PCR was used to screen 8 HHVs. RESULTS: Two thousand and fifty-two patients (49.28%) were HHV-positive among 4164 patients screened. Among these patients screened, the infection spectra of 8 human HHVs in hematological diseases as well as patients received allogeneic hematopoietic stem cell transplantation of totally 2994 patients were summarized as follows: the positive rate of EBV (29.49%) was the highest, that of HCMV (23.15%), HHV-6 was 18.77% and HHV-7 was 17.64%, while the remaining 4 HHVs all≤2.1%. The rate of co-infection of various HHVs was significantly higher than that of single infection of HHV among all these disease groups except familial hemophagocytic lymphohistiocytosis, for which single EBV infection was the most common. The differences of positive rates among these 8 human HHVs in hematological diseases as well as patients received allogeneic hematopoietic stem cell transplantation were statistically significant by Chi-square test of R*C tables (χ2=54.99, P＜0.05). For each HHV, the differences of positive rates among the above-mentioned disease groups were also statistically significant except HHV-8 (P＜0.05). CONCLUSION: The patients with various blood diseases have different activated infection spectra of HHVs. EBV, HCMV, HHV-6 and HHV-7 are most common in HHVs infection. Different HHVs infections correlate with different hematologion diseases.