An in vitro investigation of telocytes-educated macrophages: morphology, heterocellular junctions, apoptosis and invasion analysis.

BACKGROUND: Telocytes (TCs), a recently discovered novel type of interstitial cells, were also found in a wide variety of human and mammalian reproductive organs/tissues, including uterus, oviduct and placenta. Previously, we demonstrated that TCs-conditioned media was capable of activating peritoneal macrophages (pMACs) through paracrine effects. This study investigates the hypothesis that direct interaction of TCs with pMACs will also play a significant role in immunoregulation of pMACs. METHODS: TCs and pMACs were derived from the uterus and intraperitoneal cavity of female BALB/c mice, respectively. TCs were identified by immunofluorescence and then co-cultured directly with pMACs for 24 h without added cytokines, to observe the in vitro biological behavior of pMACs. We used histochemical staining to study morphology and mitochondrial metabolism of pMACs, scanning electron microscopy to study heterocellular junctions, flow cytometry to investigate mitochondrial membrane potential (ΔΨm) and apoptosis, and transwell chambers to study invasion ability. Student-t test was used accordingly. RESULTS: Presently, TCs with typical structure and immunophenotype of double CD-34-positive/vimentin-positive were successfully isolated. pMACs co-cultured with TCs showed obviously morphological activation, with enhanced energy metabolism (P < 0.05). Meanwhile, direct physical cell-to-cell interaction promoted the development of heterocellular junctions between TCs and pMACs. Furthermore, TCs treatment markedly reduced the depletion of ΔΨm in co-cultured pMACs (all P < 0.05), and inhibited their apoptosis (P < 0.05). Functionally, pMACs co-cultured with TCs showed enhanced invasion ability (P < 0.05). CONCLUSIONS: Direct physical cell-to-cell interaction promoted the development of heterocellular junctions between TCs and pMACs, presumably responsible for the observed novel efficient way of pMACs activation via mitochondrial signaling pathway. TCs-educated pMACs might be a promising way to restore the defective immunosurveillance in endometriosis (EMs), led to the enhanced treatment efficacy of EMs in a simple and clinically feasible fashion.

Identification of Matrix Metalloproteinase-2 and 9 as Biomarker of Intrahepatic Cholestasis of Pregnancy.

Intrahepatic cholestasis of pregnancy (ICP) is a severe liver disease uniquely occurring during pregnancy. In this study we aimed to identify novel biomarker for the diagnosis of ICP in Chinese population. 50 healthy pregnant women, 50 mild ICP patients and 48 severe ICP patients were enrolled for this study. Liver function tests, including serum total bilirubin, direct bilirubin, alanine transaminase, aspartate aminotransferase and cholyglycine, were performed in all participants. After an overnight fast serum levels of total bile acids (TBA), matrix metalloproteinase (MMP)-2 and MMP-9 were measured, and their correlation with liver function tests were analyzed. The observed increase in serum TBA in ICP patients was not statistically significant which made it unreliable for diagnosis of ICP in Chinese population. On the other hand, both MMP-2 and MMP-9 serum levels exhibited a progressive and significant elevation in mild and severe ICP patients compared with healthy pregnant women, which also positively correlated with liver function tests. Serum levels of both MMP-2 and MMP-9 could be reliably used as laboratory abnormalities for accurate diagnosis and sensitive grading of ICP in Chinese population.

Resveratrol reduces matrix metalloproteinases and alleviates intrahepatic cholestasis of pregnancy in rats.

Intrahepatic cholestasis of pregnancy (ICP) is a severe liver disorder occurring specifically in pregnancy, and matrix metalloproteinase (MMP)-2 and MMP-9 were found to be elevated in ICP patients. Using ethinylestradiol-induced ICP rats as the model, we examined the effect of resveratrol on ICP symptoms such as bile flow rate, serum enzymatic activities, and TBA concentration, as well as MMP levels, and compared with the known ICP drug ursodeoxycholic acid. Both MMP-2 and MMP-9 were upregulated in ICP rats, and resveratrol treatment could inhibit the elevation of both MMPs, whereas ursodeoxycholic acid did not exhibit any effect. Although ursodeoxycholic acid alleviated ICP symptoms, resveratrol treatment in general exhibited better outcome in restoring bile flow rate, serum enzymatic activities, and TBA concentration. Our results for the first instance strongly supported the potential of RE as a new therapeutic agent in treating ICP, possibly through inhibiting MMP-2 and MMP-9.

In vitro morphology, viability and cytokine secretion of uterine telocyte-activated mouse peritoneal macrophages.

Telocytes (TCs), a distinct interstitial cell population, have been identified in the uterus, oviduct and placenta, with multiple proposed potential biological functions. Their unique structure allows them to form intercellular junctions with various immunocytes, both in normal and diseased tissues, suggesting a potential functional relationship with the local immune response. It has been hypothesized that through direct heterocellular junctions or indirect paracrine effects, TCs influence the activity of local immunocytes that are involved in the inflammatory process and in immune-mediated reproductive abnormalities. However, no reliable cytological evidence for this hypothesis is currently available. In this study, we cultured primary murine uterine TCs and collected TC conditioned media (TCM). Mouse peritoneal macrophages (pMACs) were co-cultured for 48 hrs with TCM or with DMEM/F12 or lipopolysaccharide (LPS) as negative and positive controls, respectively. Normal uterine TCs with a typical structure and a CD-34-positive/vimentin-positive/c-kit-negative immunophenotype were observed during culture. Morphologically, TCM-treated pMACs displayed an obvious activation/immunoresponse, in contrast to over-stimulation and cell death after LPS treatment and no sign of activation in the presence of DMEM/F12. Accordingly, a cell counting kit 8 (CCK-8) assay indicated significant activation of pMACs by TCM and LPS compared to DMEM/F12, thus supporting the marked morphological differences among these groups of cells. Furthermore, within a panel of macrophage-derived cytokines/enzymes, interleukin-6 (IL-6) and inducible nitric oxide synthase were significantly elevated in TCM-treated pMACs; tumour necrosis factor α, IL1-R1, and IL-10 were slightly, but significantly, up-regulated; and no changes were observed for transforming growth factor-ß1, IL-1ß, IL-23α and IL-18. Our results indicate that TCs are not simply innocent bystanders but are rather functional players in the activation of pMACs; they trigger and maintain the immune response, likely through indirect paracrine effects. Thus, we provide preliminary in vitro evidence of immunoregulatory and immunosurveillance roles for TCs.

Telocytes damage in endometriosis-affected rat oviduct and potential impact on fertility.

Women with endometriosis (EMs) have unexplained infertility. The recently identified telocytes (TCs) might participate in the maintenance of structural and functional integrity of oviduct tissue, but so far the involvement of TCs in EMs-affected oviduct tissue and potential impact on fertility capacity remain unknown. By an integrated technique of haematoxylin and eosin staining, in situ immunohistochemistry and double-labelled immunofluorescence staining and electron microscopy approach, TCs were studied in the autotransplantation Sprague-Dawley rat model of EMs-affected oviduct tissue and in sham control, respectively, together with determination of iNOS, COX-2, LPO and estradiol. TCs were found in perivascular connective tissue and smooth muscle bundles in sham oviduct, with typical ultrastructural features (a slender piriform/spindle/triangular cell body, and one or more extremely long prolongations, emerged from cell bodies and extend to various directions), and specific immunophenotype of CD34-positive/vimentin-positive/c-kit-negative. However, in EMs-affected oviduct tissue (grade III), extensive ultrastructural damage (degeneration, discontinue, dissolution and destruction), significant decrease or loss of TCs and interstitial fibrosis were observed, together with elevated level of iNOS, COX-2, LPO and estradiol, thus suggestive of inflammation and ischaemia-induced TCs damage. Based on TCs distribution and intercellular connections, we proposed that such damage might be involved in structural and functional abnormalities of oviduct, such as attenuated intercellular signalling and oviduct contractility, impaired immunoregulation and stem cell-mediated tissue repair, 3-D interstitial architectural derangement and tissue fibrosis. Therefore, TCs damage might provide a new explanation and potential target for EMs-induced tubal damage and fertility disorders.

Ultrastructure damage of oviduct telocytes in rat model of acute salpingitis.

Acute salpingitis (AS) is an inflammatory disease which causes severe damage to a subset of classically described cells lining in oviduct wall and contributes to interstitial fibrosis and fertility problems. Telocytes (TCs), a newly discovered peculiar type of stromal cells, have been identified in many organs, including oviduct, with proposed multiple potential bio-functions. However, with recent increasing reports regarding TCs alterations in disease-affected tissues, there is still lack of evidence about TCs involvement in AS-affected oviduct tissues and potential pathophysiological roles. We presently identified normal TCs by their characteristic ultrastructural features and immunophenotype. However, in AS-affected oviduct tissues, TCs displayed multiple ultrastructural damage both in cellular body and prolongations, with obvious loss of TCs and development of tissue fibrosis. Furthermore, TCs lose their interstitial 3-D network connected by homocellular or heterocellular junctions between TCs and adjacent cells. And especially, TCs connected to the activated immunocytes (mononuclear cells, eosinophils) and affected local immune state (repression or activation). Meanwhile, massive neutrophils infiltration and overproduced Inducible Nitric Oxide Synthase (iNOS), COX-2, suggested mechanism of inflammatory-induced TCs damage. Consequently, TCs damage might contribute to AS-induced structural and reproductive functional abnormalities of oviduct, probably via: (i) substances, energy and functional insufficiency, presumably, e.g. TC-specific genetic material profiles, ion channels, cytoskeletal elements, Tps dynamics, etc., (ii) impaired TCs-mediated multicellular signalling, such as homeostasis/angiogenesis, tissue repair/regeneration, neurotransmission, (iii) derangement of 3-D network and impaired mechanical support for TCs-mediated multicellular signals within the stromal compartment, consequently induced interstitial fibrosis, (iv) involvement in local inflammatory process/ immunoregulation and possibly immune-mediated early pregnancy failure.

Analysis of HGF, MACC1, C-met and apoptosis-related genes in cervical carcinoma mice.

To understand the underlying pharmacological basis and the molecular mechanism of Taxol in therapy of cervical carcinoma (CC) disease, we need to explore the effect of Taxol on CC-related genes and pro-apoptosis and anti-apoptosis genes expression. Immunohistochemistry, western blot and reverse transcription-polymerase chain reaction were applied to examine postive expression levels of Bcl-2, Bax and Caspase-3, HGF, MACC1, Caspase-3 and C-met proteins and MACC1 mRNA expression in tumour of CC mice. Results showed that treatment of Taxol could increase the inhibition rate of tumour growth, positive expression levels of Caspase-3, Bax and decrease positive expression levels of Bcl-2 and Bcl-2/Bax, expression levels of HGF, MACC1 and C-met proteins and MACC1 mRNA in tumour tissue of CC mice. It can be concluded that inhibitory activity of Taxol against tumour growth in CC mice is closely associated with its modulating positive expression of Bcl-2, Bax, Caspase-3, expression of HGF, MACC1, Caspase-3 and C-met proteins and MACC1 mRNA in tumour of CC mice. In conclusion, HGF, MACC1 and C-met genes involve into malignant cervical tumors occurrence, development and prognosis, and might become potential molecular target therapy site of cervical cancer. Taxol intervention may serve as a multi-targeted CC therapeutic capable of inducing selective cancer cell death.

LOC644656 promotes cisplatin resistance in cervical cancer by recruiting ZNF143 and activating the transcription of E6-AP.

Cisplatin resistance remains a persistent challenge in cervical cancer (CC) treatment. Molecular biomarkers have garnered attention for their association with cisplatin resistance in various diseases. Long non-coding RNAs (lncRNAs) exert significant influence on CC development. This study explores the role of LOC644656 in regulating cisplatin resistance in CC. Parental and cisplatin-resistant CC cells underwent cisplatin treatment. Functional assays assessed cell proliferation and apoptosis under different conditions. RNA pull-down with mass spectrometry, along with literature review, elucidated the interaction between LOC644656, ZNF143, and E6-AP. Mechanistic assays analyzed the relationship between different factors. RT-qPCR and western blot quantified RNA and protein levels, respectively. In vivo models validated E6-AP's function. Results revealed LOC644656 overexpression in cisplatin-resistant CC cells, exacerbating cell growth. LOC644656 recruited ZNF143 to activate E6-AP transcription, promoting cisplatin resistance in CC. In conclusion, LOC644656 positively modulates E6-AP expression via ZNF143-mediated transcriptional activation, contributing to cisplatin resistance in CC.

Immunohistochemical alterations of cajal-like type of tubal interstitial cells in women with endometriosis and tubal ectopic pregnancy.

PURPOSE: The aim of the study was to observe alterations of pacemaker cells termed cajal-like type of tubal interstitial cells (t-ICC) in oviduct from early-stage EMs and tEP, discuss underlying mechanisms and potential role in tubal factor infertility (TFI). METHODS: Ten patients with early-stage EMs, 10 with unruptured tEP and 10 control subjects were included in this retrospective comparative study, received adnexectomy (salpingectomy) and/or hysterectomy. Paraffin-embedded full-thickness isthmic segment of oviduct specimens received immunohistochemistry with c-kit/CD117 antibody. Network distribution and area density of cells with features of t-ICC were analyzed. RESULTS: t-ICC was detected mainly in lamina propria and smooth muscle layers. t-ICC lost its network integrity, became less densely stained, sparse and almost invisible with relatively very rare connections in EMs and tEP, apparently differing in morphology of t-ICC from control, which demonstrated rich t-ICC immunostaining and intact network. Further quantitative analysis showed the area density of t-ICC decreased significantly in early-stage EMs and tEP compared with the control (73.9 ± 8.8 vs. 156 ± 18.3 mm(2); and 76 ± 7.4 vs. 156 ± 18.3 mm(2); both P < 0.001). CONCLUSIONS: We revealed that t-ICC underwent certain degree of cell damage, suggested that decreased expression of t-ICC network may be involved in early development of EMs and tEP, and might serve as an explanation for TFI in these patients.

PDHB-AS suppresses cervical cancer progression and cisplatin resistance via inhibition on Wnt/ß-catenin pathway.

Cervical cancer (CC) is the most prevalent gynecological malignancy occurring in the cervix. Long non-coding RNAs (lncRNAs) can act as oncogenes or anti-oncogenes in CC development. Here, we investigated the functional role and detailed mechanism of lncRNA pyruvate dehydrogenase E1 subunit beta antisense (PDHB-AS) in CC. At first, we found that PDHB-AS was significantly down-regulated in CC cells. Besides, overexpression of PDHB-AS repressed CC cell malignant behaviors. HKF-derived exosomes carried miR-4536-5p to CC cells and thereby inhibited PDHB-AS expression. Moreover, PDHB-AS inactivated the Wnt/ß-catenin pathway via impeding the nuclear translocation of ß-catenin in CC cells. In addition, miR-582-5p could bind with both PDHB-AS and Dickkopf-1 (DKK1). PDHB-AS recruited poly(A) binding protein cytoplasmic 1 (PABPC1) to inhibit Wnt7b expression. PDHB-AS interacted with RNA-binding motif protein X-linked (RBMX) to regulate cisplatin resistance in CC. Finally, we conducted in vivo experiments to confirm that HKF promoted CC tumor growth whereas PDHB-AS suppressed CC tumor growth. Collectively, PDHB-AS plays a tumor-suppressive role in the progression of CC, which suggests the therapeutic potential of PDHB-AS for CC.

Placental and maternal serum inhibin A in patients with preeclampsia and small-for-gestational-age.

AIM: The purpose of this study was to determine the levels of inhibin A simultaneously in the maternal serum and placental extract in preeclampsia (PE) with or without small-for-gestational-age (SGA) and normal controls at term, and to evaluate the relationship among changes in serum and placental inhibin A according to the severity of PE and PE with or without SGA. MATERIAL AND METHODS: This study involved 40 pregnant women; normal (n = 20), and PE (n = 20), the latter of who were classified into (i) mild (n = 10) and severe PE (n = 10); (ii) PE with SGA (n = 7) and without SGA (n = 13). Inhibin A concentrations were quantified by enzyme-linked immunosorbent assay (ELISA) in the maternal serum and placental extract. Inhibin-α subunit in the placenta was stained by immunohistochemistry (IHC), and its intensity was graded by a semiquantitative scoring method. RESULTS: There was a positive correlation in inhibin A concentrations between the serum and placental extract (r = 0.57, P < 0.001). Both maternal serum and placental inhibin A in PE groups were significantly higher than in controls, but there was no severity-dependent increase of inhibin A when compared with mild and severe PE. There was no difference in inhibin A levels between PE with and without SGA. Moreover, the inhibin-α subunit was predominantly abundant in the cytoplasm of the syncytiotrophoblasts, where the PE groups showed higher staining intensity than the controls (P < 0.000). CONCLUSION: Serum inhibin A level might be a useful biomarker for diagnosis and monitoring of PE.

[A correlative study on Bisphenol A and recurrent spontaneous abortion].

OBJECTIVE: This study was to investigate the association of Bisphenol A and unexplained recurrent spontaneous abortion. METHODS: A hospital-based 1:1 matched case-control study was conducted. Sixty patients with unexplained recurrent abortion were included. Each case was matched with one normal control by age (± 2 years), living district and the same gestational age. The levels of Bisphenol A in urine for 60 cases and 60 controls were detected using high performance liquid chromatography after fluorescent derivatization. The levels of urinary Bisphenol A in case was compared with that in control in education levels, occupation, smoking history. Data was analyzed by means of Wilcoxon-test, Student-Newman-Keuls after rank transform, univariate and multivariate conditional Logistic regression analysis. The software used was SAS 9.1.3. RESULTS: The values of urinary Bisphenol A in cases and controls were (0.10 ± 0.21) µg/ml, (0.03 ± 0.08) µg/ml, respectively. The level of urinary Bisphenol A in cases was significantly higher than that in controls (Z = 3.988, P < 0.0001). The urinary Bisphenol A levels in cases were significant higher than those in controls from senior middle school (the levels were 0.10, 0.06 µg/ml respectively, Z = 1.996, P = 0.0459), college (the levels were 0.14, 0.03 µg/ml respectively, Z = 2.586, P = 0.0097), workers or farmers (the levels were 0.08, 0.03 µg/ml respectively, Z = 2.265, P = 0.0235), businessmen (the levels were 0.10, 0.03 µg/ml respectively, Z = 2.544, P = 0.0109), and no passive smokers (the levels were 0.09, 0.03 µg/ml respectively, Z = 3.767, P = 0.0002). After adjustment by age, body mass index, marital status during pregnancy and other factors, compared to Bisphenol A below 0.06 µg/ml, the adjusted OR was 4.03 (1.67 - 9.74) for Bisphenol A levels between 0.06 µg/ml and 0.20 µg/ml, and was 5.46 (1.95 - 15.27) for Bisphenol A over 0.20 µg/ml. The risk of unexplained recurrent spontaneous abortion increased progressively with the growth of urinary Bisphenol A levels (χ(2) = 13.042, trend test P = 0.0003). There were significant differences on Bisphenol A among controls, two abortions, and three or more abortions (the levels were 0.03 µg/ml, 0.09 µg/ml, 0.21 µg/ml respectively, F = 9.04, P = 0.0002). CONCLUSION: Exposure to Bisphenol A may be associated with unexplained recurrent spontaneous abortion.

Aberrantly increased mRNA expression of betaglycan, an inhibin co-receptor in the ovarian tissues in women with polycystic ovary syndrome.

AIM: To compare the gene expressional levels of receptors for activins and inhibins in ovarian tissues between women with polycystic ovary syndrome (PCOS) and normal controls, and to analyze their biologically relevant associations with serum hormone levels. METHODS: Total RNA of ovarian tissues from PCOS (n = 14) and normal controls (n = 21) were isolated during the follicular phase of the menstrual cycle. Real-time PCRs were performed to examine the relative mRNA expression levels of the activin receptors, including activin receptor type IA (ActRIA), type IB (ActRIB), type IIA (ActRIIA), and type IIB (ActRIIB), and the inhibin receptors, betaglycan and an inhibin binding protein (InhBP/p120). At the same time, the serum levels of estradiol (E(2)), testosterone (T), follicle-stimulating hormone (FSH), luteinizing hormone (LH), inhibin B and activin A were measured. RESULTS: The PCOS patients showed endocrine characteristics with higher concentrations of LH, T and inhibin B, and a lower concentration of activin A. Real-time PCR demonstrated that the relative expression level of betaglycan against that of GAPDH was increased 1.5-fold in the ovarian tissues during the follicular phase of PCOS patients when compared with normal controls, while the activin receptors ActRIA, ActRIB, ActRIIA, ActRIIB, and the inhibin co-receptor InhBP/p120 were unchanged. Moreover, the betaglycan mRNA expression showed biologically relevant associations with serum FSH, LH, E(2) and inhibin B levels in both the PCOS and normal controls. CONCLUSIONS: This is the first report to demonstrate the aberrantly increased expression of betaglycan mRNA in PCOS ovaries. The mechanism by which betaglycan contributes to the pathologic process of PCOS remains to be clarified.

[Differences in the expression of inhibin receptors and activin receptors in normal human ovaries and their significance].

OBJECTIVE: To explore the differences in the expression of inhibin (INH) receptors and activin (ACT) receptors in the follicular/luteinic phase in normal human ovaries and their relationship with female endocrine hormone levels. METHODS: Real time PCR and immunohistochemistry were used to determine the expression of inhibin receptors (INHR) genes, activin receptors (ACTR) genes. Serum estradiol (E2), follicle stimulating hormone (FSH), luteinizing hormone (LH), INHB, ACTA levels were determined by a solid quantitative sandwich enzyme immunoassay technique (Sandwich ELISA) in 21 women during follicular phase and another 21 women during luteinic phase, the correlations between each gene and each hormone were analyzed. RESULTS: (1) ACT type I and II receptors genes (ACTR I A, ACTR I B, ACTR II A, ACTR II B) and INH receptor beta-glycan genes were expressed higher in the follicular phase than in the luteinic phase: ACTR I A (0.50 +/- 0.17 vs 0.36 +/- 0.18; P < 0.05), ACTR I B (0.050 +/- 0.019 vs 0.036 +/- 0.020; P < 0.05), ACTR II A (0.10 +/- 0.04 vs 0.07 +/- 0.04; P < 0.05), ACTR II B (0.28 +/- 0.10 vs 0.19 +/- 0.11; P < 0.05), beta-glycan (0.26 +/- 0.10 vs 0.17 +/- 0.09; P < 0.01). (2) The intensities of ACTR I A, ACTR II A, beta-glycan immunostaining in human normal ovaries in the follicular phase were significantly stronger compared to those in luteinic phase. In the follicular phase beta-glycan expression was positively correlated with serum E2, FSH, LH, INHB levels. The correlation coefficient was 0.53 (P < 0.05), 0.74 (P < 0.01), 0.85 (P < 0.01) and 0.76 (P < 0.01) respectively. CONCLUSION: In normal human ovary in the follicular phase INH and ACT bind their receptors and down-regulate or up-regulate FSH, thus influencing the follicular development.

The clinical value of HPV E6/E7 and STAT3 mRNA detection in cervical cancer screening.

OBJECTIVE: To explore the value of human papillomavirus (HPV) E6/E7 and signal transducer and activator of transcription 3 (STAT3) mRNA detection in the screening of cervical lesions. METHODS: 192 patients with abnormal ThinPrep cytology test (TCT) results and/or high-risk HPV infection were screened to identify possible cervical lesions in cases. Diagnoses were confirmed by histopathology. Fluorescence in situ hybridization (FISH) was performed to detect and qualify the mRNAs of HPV E6/E7, STAT3, and Survivin in cervical exfoliated cells. In addition, the performance of separate and combined mRNA detection methods were compared with TCT, HR-HPV DNA schemes respectively. RESULTS: 1. Compared with HPVE6/E7 and STAT3 mRNA methods, Survivin mRNA assay had poor specificity (Sp), Youden index (YI) and concordance rate. 2. HPV E6/E7, STAT3, and STAT3 + HR-HPV methods had the best Sp, concordance rate and positive predictive value (PPV) for cervical lesions screening and atypical squamous cells of undetermined significance (ASCUS) triage. For screening of high grade squamous intraepithelial lesions or greater (HSILs+), no difference was observed in the Se of mRNA detection methods in comparison with that of TCT, HR-HPV and TCT + HR-HPV, whereas the false positive rate (FPR) decreased by 41.48%/55.99%/17.19% and the colposcopy referral rate reduced by about 20.00%/25.00%/11.17%. For triage of women with ASCUS, no difference was observed in the Se of mRNA detection methods as compared to that of HR-HPV (χ2 = 1.05, P > 0.75), while the FPR decreased by 45.83%/37.50%/41.66% and the colposcopy referral rate reduced by 32.42%/22.60%/25.28%, respectively. The Se, YI, and PPV of the combined methods increased in comparison to each method alone. 3. Compared with the TCT + HR-HPV method, HPV E6/E7 + STAT3 method had perfect Sp (95.92%) and PPV (95.40%) for screening HSILs+, the FPR and colposcopy referral rate decreased by 31.06% and 22.48% respectively. CONCLUSIONS: 1. The expression of HPV E6/E7 and STAT3 mRNA confirmed using FISH assay is expected to be a new method and molecular marker for cervical lesions screening. Survivin mRNA was excluded due to its poor performance. 2. HPV E6/E7, STAT3, and STAT3 +HR-HPV assays could be new approaches for cervical cancer screening and ASCUS triage, and the efficiency of combined screening program was better than that of a separate one. 3. HPV E6/E7 + STAT3 regimen is expected to be a diagnostic strategy for cervical lesions.

HOXD-AS1 Exerts Oncogenic Functions and Promotes Chemoresistance in Cisplatin-Resistant Cervical Cancer Cells.

Long noncoding RNAs (lncRNAs) are important regulators in various human diseases. The lncRNA HOXD-AS1 is a tumor promoter in ovarian cancer, glioma, and lung cancer, but the specific effects of HOXD-AS1 on cervical cancer (CC) chemoresistance remain unclear. Here, the level of HOXD-AS1 in nonmalignant and CC tissues as well as in CC cells and cisplatin-resistant CC cells was determined. qRT-PCR indicated that HOXD-AS1 was overexpressed in CC tissues and cisplatin-resistant CC cells. Loss-of-function assays showed that downregulation of HOXD-AS1 expression suppressed chemoresistance of cisplatin-resistant CC cells. HOXD-AS1 targeted miR-130a-3p, and in gain-of-function assays miR-130a-3p could reverse cisplatin resistance of CC cells. miR-130a-3p in turn targeted zinc finger E-box homeobox 1 (ZEB1). These results collectively show that HOXD-AS1 can act as a competing endogenous RNA to upregulate ZEB1 expression via miR-130a-3p. The effects of the HOXD-AS1-miR-130a-3p-ZEB1 axis on cisplatin resistance of cisplatin-resistant CC cells were supported by rescue assay results. In summary, HOXD-AS1 enhanced chemoresistance of cisplatin-resistant CC cells by modulating miR-130a-3p/ZEB1 axis expression.

AGER promotes proliferation and migration in cervical cancer.

The receptor for advanced glycation end products (AGER) is an oncogenic transmembranous receptor up-regulated in various human cancers. We have previously reported that AGER was overexpressed in squamous cervical cancer. However, mechanisms of AGER involved in the progression of cervical cancer are unknown. In the present study, we investigated the effects of AGER on biological behavior, including proliferation, apoptosis, and migration using multiple biological approaches. AGER protein primarily localized in the cytoplasm and cytomembrane of cervical squamous cancer cells. Blockage of AGER with multiple siRNAs suppressed proliferation, stimulated apoptosis, inhibited migration of cervical squamous cancer cells. Conversely, overexpression of AGER increased cell proliferation, migration, and inhibited cell apoptosis. These results indicate that AGER promotes proliferation, migration, and inhibits apoptosis of squamous cervical cancer and might function as a tumor promoter in cervical cancer. Our study provides novel evidence for a potential role of AGER in bridging human papillomavirus (HPV)-induced inflammation and cervical cancer.

Knockdown of miR-629 Inhibits Ovarian Cancer Malignant Behaviors by Targeting Testis-Specific Y-Like Protein 5.

Ovarian cancer (OC) is the most lethal gynecological cancer. The molecular mechanism of it is complicated, and numerous researches suggest that microRNAs are key regulators for it. This study was to investigate the pivotal role of miR-629 in the progression of OC and to reveal the possible molecular mechanism of its action. Testis-specific Y-like protein 5 (TSPYL5) is a tumor suppressor gene in various cancers, but there is little for its role in OC. OC OVCAR3 cells were transfected with the miR-629 vector, miR-629 inhibitor, and/or small interfering RNA (siRNA) targeting TSPYL5 (si-TSPYL5), respectively. After transfection, cell apoptosis, the ability of migration, and invasion were explored, as well as the level of miR-629 and TSPYL5 protein expression were detected by quantitative polymerase chain reaction and western blot. Compared with the control, there was increasing of miR-629, and decreasing of TSPYL5 and caspase 3 in OC tissue. Overexpression of miR-629 promoted the cell ability of migration and invasion and reduced OC cell apoptosis. In addition, elevated cancer inhibition ability of TSPYL5 induced by the miR-629 inhibitor was significantly blocked by inhibition of TSPYL5 (si-TSPYL5). All the above results suggested that miR-629 could promote OC proliferation, migration, and invasion by directly suppressing TSPYL5 expression, and inhibition of miR-629 might serve as a therapeutic target for OC.

High expression of LAMP1 as a prognostic marker in patients with epithelial ovarian cancer.

Lysosome Associated Membrane Protein-1 (LAMP1), expressed in several functional processes, is a heavily glycosylated lysosomal membrane protein which is able to protect the lysosomal membranes from intracellular proteolysis. Its pro-tumorigenic effects are involved in the development of several types of Malignancy. However, the role of LAMP1 in human Epithelial Ovarian Cancer (EOC) patients remains unclear. To demonstrate its prognostic significance in EOC, the methods of RT-PCR and IHC were used to evaluate LAMP1 expression in both EOC and nonmalignant tissues. What's more, the relationship between LAMP1 and clinicopathological factors was investigated. Survival analysis was used to elaborate its prognostic value. Expression of LAMP1 in tumor cells in EOC was significantly higher than those in noncancerous tissues. LAMP1 over-expression in EOC was significantly related to FIGO stage, metastasis invasion, positive ascites cell and the level of serum CA125. The Kaplan-Meier plot and Cox regression method have shown that LAMP1 over-expression in EOC and FIGO stage was significantly related to malignant features in EOC and EOC patients' unfavorable survival. It was strongly evidenced in our research that LAMP1 is a prognostic factor in EOC. Our results suggest that LAMP1 could be used as a poor prognostic factor and novel therapeutic target for EOC.

Higher urinary bisphenol A concentration is associated with unexplained recurrent miscarriage risk: evidence from a case-control study in eastern China.

BACKGROUND: Evidence about the association between Bisphenol A (BPA) and the risk of recurrent miscarriage (RM) in human being is still limited. OBJECTIVE: We evaluated the association of urinary BPA concentrations with RM in human being. METHODS: A hospital-based 1:2 matched case-control study on RM was carried out in Suzhou and Kunshan in Jiangsu Province in China between August 2008 and November 2011. Total urinary BPA concentrations in 264 eligible urine samples (102 RM patients and 162 controls) were measured using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The Wilcoxon test and conditional logistic regression were used to estimate the differences between the groups and odds ratios (OR) with 95% confidence intervals (CI), respectively. RESULTS: The median ± IQR (interquartile range) (P75-P25) values of non-creatinine-adjusted total urinary BPA levels in the RM patients and the controls were 1.66 ± 3.69 ng/ml and 0.58 ± 1.07 ng/ml, respectively (0.98 ± 2.67 µg/g Cr (creatinine) and 0.40 ± 0.77 µg/g Cr. The adjusted BPA level was significantly higher in the RM patients than in the controls (Wilcoxon test, Z = 4.476, P < 0.001). Higher level of urinary BPA was significantly associated with an increased risk of RM (P-trend < 0.001). Compared to the groups with urinary BPA levels less than 0.16 µg/g Cr, the women with levels of 0.40-0.93 µg/g Cr and 0.93 µg/g Cr or above had a significantly higher risk of RM (OR = 3.91, 95%CI: 1.23-12.45 and OR = 9.34, 95%CI: 3.06-28.44) that persisted after adjusting for confounding factors. The time from recently RM date to recruitment does not significantly influence the urinary BPA level (P = 0.090). CONCLUSION: Exposure to BPA may be associated with RM risk.