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.

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.

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.

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.

[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.

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.

Sortase A-mediated modification of the Streptococcus mutans transcriptome and virulence traits.

Sortase A contributes to adhesion and biofilm formation of Streptococcus mutans by anchoring surface proteins like P1 onto the cell wall, and few other functional characterization has been annotated to this protein and its coding gene srtA. In this study we investigated that whether srtA deletion would affect S. mutans virulence determinants in addition to adhesion and further explored whether these effects were caused due to changes in S. mutans genomic transcription. We used acid-killing assays, glycolytic rate assessments, and exopolysaccharide (EPS) formation tests to detect whether srtA deletion influenced S. mutans acid tolerance/production and glucan formation. Comparisons between RNA-sequencing data from both the exponential and stationary phases of UA159 and the srtA-deleted strain were made to determine the impact of srtA knockout on S. mutans genomic transcription. Results of our assays indicated that S. mutans aciduricity was enhanced in the srtA deleted strain when bacterial cells were directly subjected to pH 2.8, but the enhancement was repressed when the acid tolerance response was induced in advance. The srtA mutation strain exhibited reduced EPS formation in mature biofilms. SrtA deletion led to pleiotropic changes in the S. mutans transcriptome with a growth phase-dependent pattern. The affected genes mainly included those involved in aciduricity, carbohydrate transport, and EPS formation. It was concluded that S. mutans srtA exhibited multiple effects on the virulence traits of this pathogen, including acid tolerance and glucan formation, and that these alterations could be partially due to transcriptional changes upon loss of srtA.

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.

[Link between sortase A function and cariogenicity of Streptococcus mutans: a preliminary metabolomics analysis].

OBJECTIVE: This study intends to explore the mechanism underlying the support of sortase A (SrtA) of the cariogenicity of Streptococcus mutans (S. mutans). METHODS: We performed a metabonomics study based on ¹H nuclear magnetic resonance spectroscopy (NMR), in which we compared the extracellular metabolites of wild-type S. mutans UA159 with those of its SrtA-deficient strain. Metabolite differences among strains were identified using a combination of principal component analysis and orthogonality partial least square discriminant analysis. RESULTS: Several differences corresponding mostly to unknown metabolites were identified. Some amino acids such as leucine and valine (δ 0.92×10⁻6-1.20×10⁻6), lactic acid ( δ1.28×10⁻6), oxoglutaric acid (δ 3.00×10⁻6), and glycine (δ 3.60×10⁻6) differed among strains. CONCLUSIONS: This work establishes the feasibility of using ¹H NMR-based metabonomics to provide leads for research into molecular factors that promote caries. The database of microbial metabolites should be also improved in further studies.

Establishment of an miR-137-knockout cell model using CRISPR/Cas9 genome editing.

MicroRNA-137 (miR-137) has been reported to be abnormally expressed in a variety of types of cancer, including ovarian cancer. However, the roles served by miR-137 in cancer are not fully understood. In the present study, 3 single guide RNAs (sgRNAs) were designed, synthesized and inserted into pXPR001 plasmids. The pXPR001-sgRNA plasmids were verified using sequencing and integrated into the genome of the ovarian cancer cell line, A2780, through lentiviral transduction, puromycin selection and single-cell culture. PCR products amplified from single-cell cultures using primers covering miR-137 targeting sites were sequenced to identify clones with miR-137 gene disruption. Genome editing was detected in 72% of the clones derived from sgRNA2, 4% from sgRNA3 and 0% from sgRNA1. Of the clones from sgRNA2, 32% contained 1 edited miR-137 allele and 40% contained 2 edited miR-137 alleles. The expression of miR-137 in clones #2 and #3 could not be detected by reverse transcription-quantitative polymerase chain reaction. In addition, an MTT assay demonstrated that clones #2 and #3 exhibited enhanced proliferation. In conclusion, an miR-137-knockout cell model was successfully established in A2780 cells using CRISPR/Cas9 technology.

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.

Effects of human adipose-derived mesenchymal stem cells combined with estrogen on regulatory T cells in patients with premature ovarian insufficiency.

OBJECTIVE: To investigate the effects of human adipose-derived mesenchymal stem cells (hADSCs) combined with estrogen on regulatory T cells (Tregs) in patients with premature ovarian insufficiency (POI). METHODS: hADSCs were isolated by enzymatic digestion and identified by flow cytometry. Peripheral blood mononuclear cells (PBMCs) were isolated from POI patients and healthy controls. PBMCs were cultured in the following experimental groups: the control group, hADSC group, estrogen group and combined group. The PBMCs in the hADSC group were co-cultured with hADSCs at concentrations of 1×104, 2×104, or 1×105 cells/well, and the estrogen group was co-cultured with 10-9, 10-8, or 10-7mol/L 17ß-estradiol. Cell proliferation was measured with the CCK-8 assay. The percentage of CD4+ CD25+ Foxp3+ Tregs was measured by flow cytometry. The expression levels of Foxp3, TGF-ß1 and IFN-γ were measured by real-time PCR. RESULTS: Treatment with hADSCs, estrogen and their combination promoted Tregs differentiation of PBMCs from POI patients and healthy controls. An increase in the percentage of CD4+ CD25+ Foxp3+ Tregs was observed when PBMCs were co-cultured with hADSCs, 17ß-estradiol and their combination. Foxp3 and TGF-ß1 mRNA expression was higher and IFN-γ mRNA expression was lower in the hADSCs, estrogen and combined groups than in the control group. CONCLUSION: Combined treatment with hADSCs and estrogen played an immunomodulatory role by promoting Tregs proliferation, thereby potentially improving impaired ovarian function.

MicroRNA-17 downregulates expression of the PTEN gene to promote the occurrence and development of adenomyosis.

The aim of the current study was to evaluate the expression of microRNA (miR)-17 in the endometrial tissues of patients with adenomyosis (AM) and determine its biological function in the occurrence and development of the disease. A total of 45 fresh endometrial tissues of AM patients and 32 normal endometrial tissues were collected from healthy controls. The expression of miR-17 was evaluated using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The miR-17-targeting gene phosphatase and tensin homolog (PTEN) was predicted using bioinformatics and its expression was evaluated with RT-qPCR and western blot analysis. Endometrial cells were isolated from patients with AM and healthy controls. They were cultured in vitro and transfected with antagomiR-17 to downregulate miR-17 expression, subsequently cell viability and apoptosis were measured using MTT and flow cytometry. The expression of PTEN and cell cycle- and apoptosis-related proteins were evaluated using western blot analysis. Endometrial cells that stably overexpressed PTEN were screened in vitro by co-culture with G418. A dual-luciferase reporter assay was conducted to verify whether miR-17 was directly bound to PTEN mRNA. The results demonstrated that expression of miR-17 was significantly increased in the endometrial tissues of patients with AM compared with control patients (P<0.05). PTEN mRNA and protein expression were significantly lower in the AM group compared with the control group (P<0.05). When the expression of miR-17 in the cells was downregulated, the expression of PTEN was significantly increased (P<0.05). In addition, expression of Bcl-2 protein was significantly decreased and that of Bax protein significantly increased compared with the negative control (both P<0.05). The expression of cyclins E1 and D1 were also significantly downregulated (P<0.05). When PTEN was overexpressed or miR-17 was downregulated, the viability of endometrial cells significantly decreased and cell apoptosis significantly increased (all P<0.05). A dual-luciferase reporter assay indicated that miR-17 could directly bind to the PTEN mRNA 3'-untranslated region to regulate its expression. Thus the current study indicates that expression of miR-17 was increased in the endometrial tissues of patients with AM and may influence cell apoptosis and cyclin expression through the targeted regulation of PTEN. These results suggest that miR-17 promotes the occurrence and development of AM.

Overexpression of miR-21 in stem cells improves ovarian structure and function in rats with chemotherapy-induced ovarian damage by targeting PDCD4 and PTEN to inhibit granulosa cell apoptosis.

BACKGROUND: Chemotherapy-induced premature ovarian failure (POF) is a severe complication affecting tumor patients at a childbearing age. Mesenchymal stem cells (MSCs) can partially restore the ovarian structure and function damaged by chemotherapy. miR-21 is a microRNA that can regulate cell apoptosis. This study discusses the repair effect and mechanism of MSCs overexpressing miR-21 on chemotherapy-induced POF. METHODS: Rat MSCs and granulosa cells (GCs) were isolated in vitro. MSCs were transfected with miR-21 lentiviral vector (LV-miR-21) to obtain MSCs stably expressing miR-21 (miR-21-MSCs). The microenvironment of an ovary receiving chemotherapy was mimicked by adding phosphamide mustard (PM) into the cellular culture medium. The apoptosis rate and the mRNA and protein expression of target genes PTEN and PDCD4 were detected in MSCs. Apoptosis was induced by adding PM into the culture medium for GCs, which were cocultured with miR-21-MSCs. The apoptosis rate and the mRNA and protein expression of PTEN and PDCD4 were detected. The chemotherapy-induced POF model was built into rats by intraperitoneal cyclophosphamide injection. miR-21-MSCs were transplanted into the bilateral ovary. The rats were sacrificed at 15, 30, 45, and 60 days after the last injection. The ovarian weights, follicle count, estrous cycle, and sex hormone levels (estradiol (E2) and follicle-stimulating hormone (FSH)) were detected. Apoptosis of GCs was determined by TUNEL assay. The miR-21 and mRNA and protein expression of PTEN and PDCD4 were determined. RESULTS: The apoptosis decreased in MSCs transfected with miR-21. The mRNA and protein expression of target genes PTEN and PDCD4 was downregulated. GCs cocultured with miR-21-MSCs showed a decreased apoptosis, an upregulation of miR-21, and a downregulation of PTEN and PDCD4. Following the injection of miR-21-MSCs, the ovarian weight and follicle counts increased; E2 levels increased while FSH levels decreased, with less severe apoptosis of GCs. The miR-21 expression in the ovaries was upregulated, while the mRNA expression and protein expression of PTEN and PDCD4 were downregulated. CONCLUSIONS: Overexpression of miR-21 in MSCs promoted efficacy against chemotherapy-induced POF and its improvement of the repair effect was related to the inhibition of GC apoptosis by targeting PTEN and PDCD4.

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.

Treatment of Early Stage Endometrial Cancer by Transumbilical Laparoendoscopic Single-Site Surgery Versus Traditional Laparoscopic Surgery: A Comparison Study.

To compare the outcomes of transumbilical laparoendoscopic single-site surgery (TU-LESS) versus traditional laparoscopic surgery (TLS) for early stage endometrial cancer (EC).We retrospectively reviewed the medical records of patients with early stage EC who were surgically treated by TU-LESS or TLS between 2011 and 2014 in a tertiary care teaching hospital. We identified 18 EC patients who underwent TU-LESS. Propensity score matching was used to match this group with 18 EC patients who underwent TLS.All patients underwent laparoscopic-assisted vaginal hysterectomy, bilateral salpingo-oophorectomy, and systematic pelvic lymphadenectomy by TU-LESS or TLS without conversion to laparoscopy or laparotomy. Number of pelvic lymph nodes retrieved, operative time and estimated blood loss were comparable between 2 groups. Satisfaction values of the cosmetic outcome evaluated by the patient at day 30 after surgery were significantly higher in TU-LESS group than that in TLS group (9.6 ±â€Š0.8 vs 7.5 ±â€Š0.7, P < 0.001), while there was no statistical difference in postoperative complications within 30 days after surgery, postoperative hospital stay, and hospital cost.For the surgical management of early stage EC, TU-LESS may be a feasible alternative approach to TLS, with comparable short-term surgical outcomes and superior cosmetic outcome. Future large-scale prospective studies are needed to identify these benefits.

MicroRNA-29b inhibits TGF-ß1-induced fibrosis via regulation of the TGF-ß1/Smad pathway in primary human endometrial stromal cells.

Transforming growth factor (TGF)­ß1 has a key role in the regulation of fibrosis and organ dysfunction. During the pathogenesis and progression of vital organ fibrosis, the microRNA (miR)­29 family is irregularly downregulated and exogenous supplementation of miR­29b has a strong anti­fibrotic capacity. However, whether TGF­ß1 is able to provoke endometrial fibrosis, and the role of miR­29 in endometrial fibrosis remain unclear. In the present study, RT­qPCR, immunocytochemistry, western blot analysis, scanning electron microscopy, immunofluorescence staining, cell proliferation assay and flow cytometric analysis were employed. The results demonstrated that the expression levels of collagen, type 1, alpha 1 (COL1A1), α­smooth muscle actin (α­SMA) and phosphorylated (p)­Smad2/3 were increased, whereas miR­29b and maternally expressed gene 3 (MEG3) were decreased in primary endometrial stromal cells (ESCs) in response to TGF­ß1 stimulation, in a time and dose­dependent manner. Furthermore, overexpression of miR­29b markedly reduced the expression levels of COL1A1 and α­SMA, and decreased the expression and nuclear accumulation of p­Smad2/3. In addition, ectopic overexpression of miR­29b increased the expression levels of MEG3, inhibited myofibroblast­like cell proliferation and induced apoptosis. These findings indicated that miR­29b may have a significant anti­fibrotic role, and may attenuate TGF­ß1­induced fibrosis in ESCs. Therefore, exogenous miR­29b may serve as a potential therapeutic agent for the treatment of endometrial fibrosis.

Interceed and Estrogen Reduce Uterine Adhesions and Fibrosis and Improve Endometrial Receptivity in a Rabbit Model of Intrauterine Adhesions.

Intrauterine adhesions (IUA) remain a major cause of infertility. Interceed, a regenerated cellulose adhesion barrier, is used to prevent adhesions in abdominal cavity. This study aimed to determine whether Interceed could reduce adhesions and tissue fibrosis and improve endometrial receptivity (ER) in rabbit. Rabbits were randomized into 6 groups: sham operation, Interceed control, IUA model, Interceed therapy, estrogen therapy, and combination therapy. Four rabbits per group were euthanized to evaluate adhesion severity on the day before intervention and day 7, 14, and 28 after intervention. Number of endometrial glands and degree of endometrial fibrosis acted as markers for adhesion severity. Pseudopregnancy was induced in the remainder, and 8 rabbits per group were killed for assessing ER on days 6, 7, and 8 of pseudopregnancy by ανß3 integrin and pinopode. We found that Interceed or estrogen therapy led to significant improvement in the adhesion severity on day 28 after intervention, respectively, compared to IUA model group (all P < .05). However, after combination therapy, such improvement achieved comparable to sham operation group as early as day 14 after intervention (glands, P = .711, fibrosis, P = .154). Among the IUA models treated, ER was highest after combination therapy on day 7 of pseudopregnancy, similar to sham operation group (integrin, P = .352, pinopode, P = .154). In conclusion, Interceed and estrogen reduce adhesions and tissue fibrosis and improve ER in a rabbit model and may be novel therapeutic approaches for infertility resulting from IUA.

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.

MicroRNA-29b Inhibits Endometrial Fibrosis by Regulating the Sp1-TGF-ß1/Smad-CTGF Axis in a Rat Model.

Intrauterine adhesions (IUAs), which are characterized by endometrial fibrosis, increase the risk of secondary infertility and recurrent miscarriage. MicroRNA-29 (miR-29) is a potent inhibitor of TGF-ß1/Smad signaling. In this study, we investigated the therapeutic potential of agomir-29b, an miR-29b mimic, in endometrial fibrosis induced by dual injury (uterine curettage and lipopolysaccharide treatment) in a rat model of IUA and explored the underlying mechanism. We found that injured rats developed endometrial fibrosis characterized by increased COL1A1 and α-smooth muscle actin expression and decreased E-cadherin expression, associated with a loss of miR-29b. Overexpression of miR-29b before injury prevented endometrial fibrosis including collagen accumulation and epithelial-mesenchymal transition. Delay of agomir-29b treatment until endometrial fibrosis was established on day 4 also halted the progression of disease. Further experiments indicated that miR-29b inhibited endometrial fibrosis via blockade of the Sp1-TGF-ß1/Smad-CTGF pathway. In conclusion, agomir-29b may act as a novel and effective therapeutic agent against IUAs.