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An ultrasensitive electrochemical immunosensor was developed for detection of the cervical squamous cell carcinoma antigen (SCCA) biomarker using graphene-chitosan nanocomposites. Graphene oxide (GO) synthesized by a modified Hummer’s method was functionalized with chitosan biopolymer (CS) to produce CS-GO nanocomposites. Characterization by SEM, FTIR, Raman, XPS confirmed successful grafting of CS onto GO by electrostatic interactions. The CS-GO composite exhibited improved conductivity and biocompatibility. After optimizing the GO:CS ratio to 1:1, the nanocomposite was deposited on glassy carbon electrodes (GCE). Anti-SCCA antibodies were then immobilized covalently using pyrenebutyric acid crosslinker. The immunosensor fabrication was analyzed by EIS, CV, and DPV electrochemical techniques. The conditions including CS-GO ratio, antibody concentration, incubation time, pH, and voltage parameters were systematically optimized using orthogonal experimental design. Under optimal conditions, the immunosensor showed a linear detection range from 1 pg/mL to 100 ng/mL with a sensitivity of 196 μA/(ng/mL/cm2). The low limit of detection of 1 pg/mL was attributed to the high surface area and conductivity of the CS-GO transducer. Analysis of human serum samples spiked with SCCA demonstrated the reliability of the immunosensor for clinical applications. The graphene-nanocomposite electrochemical biosensing platform provides a rapid, low-cost, and ultrasensitive alternative to ELISA for SCCA detection in cervical cancer diagnostics.

The mortality and morbidity rates of ovarian cancer (OC) are high, but the underlying mechanisms of OC have not been characterized. In this study, we determined the role of Rho GTPase Activating Protein 30 (ARHGAP30) in OC progression. We measured ARHGAP30 abundance in OC tissue samples and cells using immunohistochemistry (IHC) and RT-qPCR. EdU, transwell, and annexin V/PI apoptosis assays were used to evaluate proliferation, invasiveness, and apoptosis of OC cells, respectively. The results showed that ARHGAP30 was overexpressed in OC tissue samples and cells. Inhibition of ARHGAP30 suppressed growth and metastasis of OC cells, and enhanced apoptosis. Knockdown of ARHGAP30 in OC cells significantly inhibited the PI3K/AKT/mTOR pathway. Treatment with the PI3K/AKT/mTOR pathway inhibitor buparlisib simulated the effects of ARHGAP30 knockdown on growth, invasiveness, and apoptosis of OC cells. Following buparlisib treatment, the expression levels of p-PI3K, p-AKT, and p-mTOR were significantly decreased. Furthermore, buparlisib inhibited the effects of ARHGAP30 upregulation on OC cell growth and invasiveness. In conclusion, ARHGAP30 regulated the PI3K/AKT/mTOR pathway to promote progression of OC.

Abstract Background Our previous two-dimensional electrophoresis experiment showed that the expression of LASP1 in patients with endometriosis was significantly higher than that of control endometrium. However, the molecular mechanism by which LASP1 is regulated in endometriosis/adenomyosis is unknown. Methods Herein, qPCR was performed to analyze the expression levels of LASP1 and miR-218-5p between endometriosis (Ems) cells and control cells. Fluorescence in situ hybridization was carried out to measure the expression level of miR-218-5p in ectopic endometrium versus normal endometrium. After miR-218-5p mimic or inhibitor were transfected, the transwell experiment was carried out to see the effect of miR-218-5p on the migration of endometrial stromal cells (ESCs). EdU was used to measure cell proliferation rate. Dual-luciferase reporter assay was used to verify the binding of hsa-miR-218-5p to the 3’UTR of LASP1. Western blot and immunofluorescence analysis were carried out to identify the protein expression pattern of LASP1 and EMT markers in endometrial tissue. Results The miR-218-5p is mainly secreted from blood vessels and expressed in the muscle layer around the endometrium, which inhibits the expression level of LASP1 by binding the 3’UTR region of LASP1 in normal ESCs. Overexpression of miR-218-5p impedes the epithelial-to-mesenchymal transition (EMT) and prevents the migration of ESCs and the expression of Vimentin in Ems. Conclusions Our findings revealed that miR-218-5p in endometrial microenvironment prevents the migration of ectopic endometrial stromal cells by inhibiting LASP1.

Abstract Background Given the important roles of the receptor-mediated lysophosphatidic acid (LPA) signaling in both reproductive tract function and gynecological cancers, it will be informative to investigate the potential role of LPA in the development of adenomyosis. The objective of this study was to evaluate the levels of LPA in plasma and the expression of six LPA receptors in the endometrial tissue collected from women with and without adenomyosis. Methods Plasma and endometrial tissue samples were collected form women with and without adenomyosis. The levels of LPA in plasma were determined by using high-performance liquid chromatography electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS). Immunohistochemistry was performed to evaluate the expression of six LPA receptors (LPA1–6) in endometrial tissue samples. The effects of LPA on IL-8 production, VEGF production and cell proliferation in human endometrial stromal cells (ESCs) were also assessed. Results LPA1 staining was localized to the cytoplasm, membrances of the epithelial cells of the endometrial glands, and there was little staining in the stromal cells. LPA2–5 staining were localized to the nuclei of stromal and glandular cells. Plasma levels of LPA were increased in adenomyosis. LPA1, LPA4 and LPA5 immunoreactivity were significantly higher in the adenomyosis group than in the control group, while LPA2 and LPA3 immunoreactivity were significantly lower in the adenomyosis group than in the control group. LPA6 was undetectable in the endometria. LPA induced the release of IL-8 from ESCs but did not affect cell proliferation and VEGF production. Conclusion These results indicate that elevated plasma levels of LPA and aberrant expression of LPA receptors in the endometria may be associated with the development of adenomyosis.

Purpose: To investigate the effect of astragaloside IV administration on the inflammatory response in endometriosis and the underlying mechanism of action. Methods: Mice were divided into two groups: endometriosis (EMs) mice and control mice (n = 12). EMs induction in mice was achieved by transplantation of mouse uterine tissue. The same procedure was performed in control mice except that a separate suture was inserted instead of endometrial tissue. After 5 weeks, EMs mice were treated with or without astragaloside IV (AIV). The tissue lesions in EMs and control mice were stained with hematoxylin and eosin staining. The activation of toll-like receptor 4 (TLR4) and nuclear factor-κB (NF-κB) p65 signaling was evaluated by western blot, while expression of inflammatory cytokines was evaluated by quantitative real-time-polymerase chain reaction (qRT-PCR), and enzyme-linked immunosorbent assay (ELISA). Results: Astragaloside IV repressed the inflammation of murine Ems lesions, and also dampened the activation of TLR4/NF-κB signaling in vivo and vitro (p < 0.01 and p < 0.001, respectively). In addition, the expression levels of inflammatory cytokines (IL-1β, IL-6, Ccl-2, and TNF-α) decreased following AIV treatment in vivo. Conclusion: The results indicate that TLR4/NF-κB signaling pathways are closely related to the inhibition of Ems inflammation by astragaloside IV. Thus, astragaloside IV may be a novel drug for the prevention and treatment of endometrioses. [ABSTRACT FROM AUTHOR]

Introduction: There has a close relationship between the balance of T helper 17 (Th17) cells and Foxp3+ regulatory T cells (Treg) and unexplained recurrent pregnancy loss (URPL). The present study is to investigate the regulatory effect of daphnetin, which is extracted from Daphne odora Var, on the balance of Th17 and Treg cells in the peripheral blood mononuclear cells (PBMCs) from patients with URPL. Material and methods: PBMCs were isolated from 35 pregnant women with URPL and 35 women with normal early pregnancies, respectively and treated with daphnetin for three days. Flow cytometry was performed to measure the proportions of Th17 and Treg cells. The level of expression of IL-6, TGF-β1 and IL-2 were detected using enzyme-linked immunosorbent assay (ELISA) and the level of expression of FoxP3, RORγt, signal transducers and activators of transcriotion 3 (STAT3) and STAT5 were detected by RT-PCR. Results: The concentrations of Th17-type cytokines IL-2 were significantly decreased in the URPL group after treatment (p < 0.01). Treg-type cytokines such as TGF-β1 and IL-6 were significantly increased after treatment (p < 0.01). At the same time, daphnetin may induce a decrease in the ratio of RORγt to Foxp3 and a Treg cell bias, which would be beneficial for pregnancy maintenance. Futhermore the expression level of STAT3 were higher in the URPL patients whereas STAT5 were lower than those in the control subjects. Conclusions: In conclusion, daphnetin may have regulatory effect on the balance of Th17 and Treg cells in the peripheral blood mononuclear cells from patients with URPL.. [ABSTRACT FROM AUTHOR]