Downregulation of HS6ST2 Inhibits Cervical Cancer Cell Migration and Invasion in Vivo and in Vitro.

BACKGROUND: Emerging evidence indicates the importance of heparan sulfate 6-O-sulfotransferase 2 (HS6ST2) in a number of developmental processes. Little is known regarding its biological function in regulating cervical cancer (CC) progression. In this study, we aim to explore the role of HS6ST2 in CC progression. METHODS: The transcriptome sequencing data of CC tissues from three databases, GSE64217, GSE138080, and GSE63514, was examined for genes with significant changes. The expression profile for HS6ST2 within CC tissue was then assessed through fluorescence quantitative PCR and immunohistochemistry and compared to data from patients with clinicopathological features. A multivariate survival analysis was performed using the COX regression. The real-time quantitative PCR assessed the HS6ST2 expression profile within CC cellular cultures. The results of knocking down HS6ST2, considering the proliferative activity and invasiveness of CC cultures in vitro, were detected through cell viability assay, clonogenic assessment, tumorsphere formation analysis, 3D invasion experiment and transwell assay. The impact of HS6ST2 knockdown in CC proliferation was also evaluated in vivo using a nude mice model. RESULTS: HS6ST2 was severely upregulated within CC tissues across the three explored databases (GSE64217, GSE138080, and GSE63514). Fluorescent quantitative PCR and immunohistochemistry experiments identified HS6ST2 as highly upregulated within patients CC tissues. Survival analysis taking into account the parameters of lymph node metastasis, Federation of Gynecology and Obstetrics (FIGO) stage, depth of invasion, pathological grade, and HS6ST2 expression level demonstrated that individuals with downregulated HS6ST2 exhibited considerably extended progression-free survival (PFS) and overall survival (OS) in comparison to upregulated HS6ST2 cases. According to the findings of COX univariate analysis, the parameters lymph node metastasis, FIGO stage, depth of invasion, pathological grade, and HS6ST2 expression level, all showed a statistically significant correlation with effect upon prognosis of CC patients. The FIGO stage, depth of invasion and expression level of HS6ST2 were identified as independent risk variables influencing CC case prognosis within subsequent COX multivariate analysis. Cell function experiments proved that HS6ST2 knockdown can considerably diminish the proliferative potential, stemness and invasive traits of CC cells. Tumor formation experiments in nude mice in vivo demonstrated that knocking down HS6ST2 can significantly thwart CC cellular proliferative properties within animal models. CONCLUSIONS: The clinicopathological features and the survival time of the patients significantly correlate with the level of HS6ST2 expression in CC tissue samples.

FBXO22 inhibits proliferation and metastasis of cervical cancer cells by mediating ubiquitination-dependent degradation of GAK.

Cervical cancer is one of the recognized malignant tumors of female reproductive system. At present, the research and development of biomarkers has attracted increasing attention, and the wide application of clinical cervical cancer screening strategies has significantly reduced its morbidity and mortality. A member of the F-box protein family, FBXO22, is involved in cell cycle, DNA damage repair and many other processes. Dysregulation of FBXO22 plays an important role in the occurrence and development of various tumors, including ovarian cancer, liver cancer and lung cancer. Nevertheless, the effect of FBXO22 in cervical cancer needs further investigation. We found that FBXO22 inhibited cervical cancer cell proliferation, migration and invasion. The results of proteomics studies suggested FBXO22 appears to target the Cyclin G Associated Kinase (GAK) for degradation. The combined results of analysis of cultured cells with altered abundance of FBXO22 by depletion or over-expression in the presence or absence of proteasomal inhibitor, comparison of protein decay rate, as well as cellular ubiquitination, support a hypothesis that FBXO22 mediates the ubiquitin-dependent degradation of GAK. Taken together, our data suggest that FBXO22 has a protective role in cervical cancer.

TRIM16 E121D variant affects the risk and prognosis of hepatocellular carcinoma by modulating the Wnt/ß-catenin pathway.

TRIM16 has been identified as a tumor suppressor in hepatocellular carcinoma (HCC). This study aimed to investigate whether there are genetic variants in TRIM16 influencing HCC risk and/or prognosis and explore the mechanisms. We performed a gene-wide single-nucleotide polymorphism (SNP) mining in TRIM16. The associations of SNPs with both HCC risk and prognosis were assessed through two independent cohorts respectively. Functional experiments were performed to investigate the underlying mechanisms. A missense variant rs2074890 (G > T, resulting in an amino acid substitution from glutamate to aspartate at code 121, E121D) of TRIM16 was found to be associated with both HCC risk (odds ratio = 0.806, p = 0.023) and prognosis (hazard ratio = 0.44, p = 0.034). Compared to the rs2074890 G allele (corresponding to TRIM16121E ) homozygote carriers, the rs2074890 T allele (corresponding to TRIM16121D ) carriers showed lower HCC risk and better overall survival. Mechanistically, TRIM16121D has stronger ability to inhibit proliferation, migration, and invasion of HCC cells. Furthermore, TRIM16121D could bind to ß-catenin better and mediate K48-linked ubiquitination to degrade ß-catenin, which leads to inhibition of Wnt/ß-catenin pathway. In conclusion, TRIM16 E121D variant impacts both risk and prognosis of HCC via regulation of Wnt/ß-catenin pathway, which may lead to better understanding the pathogenesis of HCC.

Long non-coding RNA RMST serves as a diagnostic biomarker in patients with carotid artery stenosis and predicts the occurrence of cerebral ischemic event: A retrospective study.

OBJECTIVES: The purpose of this retrospective study is to explore the diagnostic and prognostic roles of serum RMST in carotid artery stenosis (CAS). METHODS: Serum levels of RMST were detected in CAS patients, and the relationship between degree of carotid stenosis and RMST levels was analyzed. The ROC curve was drawn to evaluate RMST value in predicting the risk of CAS. Then, all CAS patients received a 5-year follow-up. K-M curve was used to analyze the significance of RMST on prognosis of CAS patients. Multi-factor cox logistic regression analysis was conducted to evaluate independent factors for outcome of CAS patients. RESULTS: An increased RMST expression was certified in CAS patients when compared with healthy controls. The increase of serum RMST expression was related to high degree of carotid stenosis. In addition, serum RMST was a possible diagnosis and an independent influencing factor of prognosis in patients with CAS. CONCLUSIONS: Raised serum RMST level was found in patients with CAS. Detecting RMST expression levels was of high value for predicting the occurrence and outcomes in CAS.

Circular RNA circ_0004488 Increases Cervical Cancer Paclitaxel Resistance via the miR-136/MEX3C Signaling Pathway.

Circular RNAs have been proven to play a pivotal role in cervical cancer development, progression, and treatment resistance. However, it is unclear how these RNAs influence chemoresistance in cervical cancer, particularly cancer stem cell (CSC)-like properties. In this study, we found that circRNA circ_0004488 was highly expressed in CSC-enriched subsets of cervical cancer cell lines. The expression of circ_0004488 was upregulated in cervical cancer cells that were resistant to paclitaxel. When circ_0004488 expression was high, the prognosis was poor. Specifically, we discovered that knocking down circ_0004488 greatly decreased the development of cervical cancer cells in vivo by decreasing cell proliferation, invasion, and sphere formation. By blocking cir_0004488, cervical cancer cells become more sensitive to paclitaxel. In cervical cancer cells, circ_0004488 acted as a microRNA-136 (miR-136) sponge, increasing the expression of MEX3C (a direct target gene of miR-136) using dual-luciferase reporter assays. Moreover, MEX3C downregulation significantly reduced cell proliferation, invasion, sphere formation, and paclitaxel resistance. In conclusion, circ_0004488 was shown to induce CSC-like features and paclitaxel resistance through the miR-136/MEX3C axis. Therefore, circ_0004488 might be a good therapeutic target for treating cervical cancer.

The Clinical Significance of Lncrna GAS5 And Mir-222-3p in Carotid Artery Stenosis.

BACKGROUND: Carotid artery stenosis (CAS) is the major pathogen of cerebral infarction and brain death. Early detection and risk prediction could help the diagnosis and improve the outcome of patients. The clinical significance of lncRNA GAS5 (GAS5) and miR-222-3p in the diagnosis and prognosis of CAS was evaluated in this study to explore novel effective biomarkers of CAS. METHODS: A total of 72 CAS patients and 63 healthy individuals (control) were enrolled in this study. The expression levels of GAS5 and miR-222-3p in study subjects were detected using PCR. The ROC, Kaplan-Meier, and Cox regression analyses were carried out to estimate the diagnostic and prognostic value of GAS5 and miR-222-3p in CAS. The interaction between GAS5 and miR-222-3p was disclosed by the dual-luciferase reporter. RESULTS: The reduced expression of GAS5 and elevated expression of miR-222-3p were observed in CAS patients compared with the healthy controls, and a significant correlation between their expression levels in CAS was revealed. GAS5 and miR-222-3p could discriminate CAS patients from the healthy controls with high sensitivity and specificity. The GAS5 downregulation and miR-222-3p upregulation could predict the poor prognosis of CAS patients and may be associated with the severe development of patients. In human vascular smooth muscle cells, miR-222-3p could negatively regulate the luciferase activity of GAS5. CONCLUSION: Both GAS5 and miR-222-3p served as the diagnostic and prognostic biomarkers of CAS. The function of GAS5 might result from the regulation of miR-222-3p, which needs further validation.

Oncogenic circular RNA circ_0007534 contributes to paclitaxel resistance in endometrial cancer by sponging miR-625 and promoting ZEB2 expression.

Circular RNAs (circRNAs) and epithelial to mesenchymal transition (EMT) have been implicated in the development of human cancer and paclitaxel resistance. CircRNA circ_0007534 has been described as a key oncogenic circular RNA that is upregulated in a variety of cancer tissues. However, whether circ_0007534 causes EMT and paclitaxel resistance in endometrial cancer is still unknown. In this work, we revealed that circ_0007534 levels were significantly higher in endometrial cancer tissues, and that high circ_0007534 expression was associated with poor differentiation, advanced tumor stage, cancer invasion, cancer metastasis, and poor prognosis in endometrial cancer patients. Overexpression of circ_0007534 boosted endometrial cancer cell proliferation, invasion, EMT, and paclitaxel resistance. Knockdown of circ_0007534 restored paclitaxel sensitivity and reversed EMT in endometrial cancer cells. We also showed that circ_0007534 enhanced endometrial cancer aggressiveness, progression, and paclitaxel resistance by sponging microRNA-625 (miR-625) and subsequently increasing the expression of the miR-625 target gene ZEB2. Our cell functional studies demonstrated that inhibiting miR-625 or increasing ZEB2 mimicked the effects of circ_0007534 overexpression. Consequently, our data show that circ_0007534 plays a crucial role in EMT and paclitaxel resistance through miR-625/ZEB2 signaling. Targeting the circ_0007534/miR-625/ZEB2 pathway might be an effective strategy for overcoming paclitaxel resistance in endometrial cancer.

High-intensity focused ultrasound ablation combined with transcatheter arterial chemoembolization improves long-term efficacy and prognosis of primary liver cancer.

BACKGROUND: To investigate the clinical efficacy of high-intensity focused ultrasound (HIFU) combined with transcatheter arterial chemoembolization (TACE) in the treatment of primary liver cancer (PLC) and its effect on the prognosis of patients. METHODS: A total of 132 patients with PLC admitted to our hospital were selected for the study, among whom 68 patients received TACE combined with HIUF and were assigned to the observation group (OG), whereas the remaining 54 patients were treated with TACE alone and were assigned to the control group (CG). The factors influencing the patients' prognosis were also evaluated by multivariate analysis. RESULTS: The total effective rate of the OG was 83.82%, which was significantly higher than that of 55.56% of the CG (P < .05). No significant difference was found in incidence of adverse reactions between the two groups (P > .05). After treatment, the increases of CD3+, CD4+, CD4+/CD8+, and NK cells in the OG were more significant than those in the CG (P < .05). However, the decrease of CD8+ cells was more significant in the OG than that in the CG (P < .05). The 3-year survival rate of patients in the OG was 61.76%, which was significantly higher than that of 40.74% in the CG (P < .05). CONCLUSION: The application of TACE combined with HIFU is effective in treating PLC, which can prolong the life expectancy and improve the prognosis of patients with PLC without increasing the incidence of adverse reactions.

TP53 R249S mutation detected in circulating tumour DNA is associated with Prognosis of hepatocellular carcinoma patients with or without hepatectomy.

BACKGROUND AND AIMS: Somatic mutation R249S in TP53 is highly common in hepatocellular carcinoma (HCC). We aim to investigate the effects of R249S in ctDNA on the prognosis of HCC. METHODS: We analysed three cohorts including 895 HCC patients. TP53 mutation spectrum was examined by direct sequencing of genomic DNA from tissue specimens in HCC patients with hepatectomy (Cohort 1, N = 260). R249S and other recurrent missense mutations were assessed for their biological functions and associations with overall survival (OS) and progression-free survival (PFS) of HCC patients in Cohort 1. R249S within circulating tumour DNA (ctDNA) was detected through droplet digital polymerase chain reaction (ddPCR) and its association with OS and PRS was analysed in HCC patients with (Cohort 2, N = 275) or without (Cohort 3, N = 360) hepatectomy. RESULTS: In Cohort 1, R249S occupied 60.28% of all TP53 mutations. Overexpression of R249S induced more serious malignant phenotypes than those of the other three identified TP53 recurrent missense mutations. Additionally, R249S, but not other missense mutations, was significantly associated with worse OS (P = .006) and PFS (P = .01) of HCC patients. Consistent with the results in Cohort 1, HCC patients in Cohorts 2 and 3 with R249S had worse OS (P = 8.291 × 10-7 and 2.608 × 10-7 in Cohorts 2 and 3, respectively) and PFS (P = 5.115 × 10-7 and 5.900 × 10-13 in Cohorts 2 and 3, respectively) compared to those without this mutation. CONCLUSIONS: TP53 R249S mutation in ctDNA may serve as a promising prognosis biomarker for HCC patients with or without hepatectomy.

Serum heart-type fatty acid-binding protein as a predictor for the development of sepsis-associated acute kidney injury.

Background: We analyzed the correlation between heart-type fatty acid-binding protein (HFABP) and the development of acute kidney injury (AKI) in patients with sepsis and estimated the predictive capacity of HFABP for sepsis-associated acute kidney injury (SAKI). Methods: In this retrospective observational study, we screened 2,452 patients who received the HFABP test in the emergency department. 442 admitted patients with sepsis were finally enrolled. Based on the diagnostic criteria for AKI in Kidney Disease: Improving Global Outcomes (KDIGO) guidelines, patients were divided into the no-AKI group (n = 317) and AKI group (n = 125). We analyzed the correlation between HFABP and SAKI occurrence by logistic regression analysis and evaluated the predictive ability of HFABP to SAKI using c-index, net reclassification improvement index (NRI) and integrated discrimination improvement index (IDI). Results: Patients in the AKI group with significantly higher the level of HFABP and in-hospital mortality. HFABP concentration is an independent risk factor for SAKI (OR: 11.398; 95% CI: 6.218-20.891, P < 0.001), but not for in-hospital mortality (OR: 1.189, 95%CI: 0.954-2.607, P = 0.076). The addition of HFABP to the prediction model significantly improved the ROC area (0.867 vs 0.755, P < 0.001), NRI 25.03% (95% CI 9.72-38.51%) and IDI 14.33 (95% CI 11.04-17.62). Conclusion: Serum HFABP is correlated with SAKI development and could become a potential predictive biomarker.

Gefitinib induces non-small cell lung cancer H1650 cell apoptosis through downregulating tumor necrosis factor-related apoptosis-inducing ligand expression levels.

Non-small cell lung cancer (NSCLC) presents severe threats to the lives of patients. Gefitinib is one of the first-line drugs available for the treatment of NSCLC in the clinical setting. The present study investigated the effects of gefitinib on NSCLC H1650 cell viability and apoptosis via MTT assays and flow cytometry. Western blot analysis was employed to detect tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) expression levels in H1650 cells. In the present study, H1650 cells were treated with TRAIL siRNA or an empty plasmid vector control, followed by gefitinib treatment to investigate apoptosis. Gefitinib treatment markedly inhibited H1650 cell viability, induced apoptosis and reduced TRAIL expression levels. TRAIL interference enhanced H1650 cell apoptosis induced by gefitinib. TRAIL overexpression suppressed gefitinib-induced H1650 cell apoptosis. In addition, gefitinib induced NSCLC H1650 cell apoptosis by downregulating TRAIL expression levels.

Epigallocatechin gallate attenuates overload­induced cardiac ECM remodeling via restoring T cell homeostasis.

It has previously been demonstrated that Epigallocatechin gallate (EGCG) has regulatory effects on cellular immunity. The present study explored whether EGCG inhibits the overload­induced cardiac extracellular matrix (ECM) remodeling through targeting the balance of T cell subpopulations. Sprague­Dawley rats were subjected to either transverse aortic constriction (TAC) or sham operation. TAC rats were treated with EGCG or valsartan (Val) for 6 weeks. The administration of EGCG or Val ameliorated the overproduction of cardiac collagen, inhibited matrix metalloproteinase (MMP) activity, decreased the expression of tissue inhibitor of MMP­2, atrial natriuretic peptide and brain natriuretic peptide. EGCG regulated the population of effector T cells and naïve T cells, restored the balance of T helper (Th) cell 17/regulatory T cells, via modulating the downstream regulator signal transducer and activator of transcription (STAT3) and STAT5. Furthermore, the ratio of interferon­Î³/interleukin (IL)­10 which indicates the balance of Th1/Th2, was restored by the treatments at varying degrees. EGCG and Val administration rescued IL­7 production, and decreased the level of IL­15 in TAC rats. EGCG has positive therapeutic potential in inhibiting cardiac ECM remodeling. Regulation of the balance of T lymphocyte subsets may be one of the underlying mechanisms responsible for this effect.

MicroRNA-134 reverses multidrug resistance in human lung adenocarcinoma cells by targeting FOXM1.

Multidrug resistance (MDR) is the primary barrier to the success of chemotherapy for lung adenocarcinoma. MicroRNA (miR)-134, which is downregulated in lung adenocarcinoma, influences cell proliferation, apoptosis and invasion of lung adenocarcinoma. However, the function of miR-134 in the MDR of lung adenocarcinoma remains unclear. In the present study, it was identified that miR-134 expression is significantly downregulated in A549/cisplatin MDR lung adenocarcinoma cells, as compared with A549 parental cells. miR-134 regulates the sensitivity of lung adenocarcinoma cells to certain anticancer drugs. Furthermore, it was demonstrated that forkhead box M1 and multidrug resistance-associated protein 1 are functional targets of miR-134. These data revealed an important role for miR-134 in the regulation of MDR in lung adenocarcinoma.

miR-326 reverses chemoresistance in human lung adenocarcinoma cells by targeting specificity protein 1.

Cisplatin resistance is a major obstacle in the treatment of lung adenocarcinoma (LAD), and its mechanism has not been fully elucidated. Here, we report that miR-326 is downregulated in cisplatin-resistant A549/CDDP cells compared with parental A549 cells. Overexpression of miR-326 reversed cisplatin chemoresistance of LAD cells in vitro and in vivo. Moreover, we identified the specificity protein 1 (SP1) gene as a novel direct target of miR-326. Knockdown of SP1 revealed similar effects as that of ectopic miR-326 expression. Decreased miR-326 expression was also detected in tumor tissues sampled from LAD patients treated with cisplatin-based chemotherapy and was proved to be correlated with high expression of SP1 and decreased sensitivity to cisplatin. Furthermore, we show that the long noncoding RNA HOTAIR repression reverses chemoresistance of LAD cells partially through modulation of miR-326/SP1 pathway. In summary, we unveil a branch of the HOTAIR/miR-326/SP1 pathway that regulates chemoresistance of LAD cells.

LncRNA MALAT1 exerts oncogenic functions in lung adenocarcinoma by targeting miR-204.

Accumulating evidence indicates that the lncRNAs play a critical role in cancer progression and metastasis. In this study, we found that MALAT1 upregulation was associated with larger tumor size and lymph-node metastasis, and also correlated with shorter overall survival of lung adenocarcinoma patients. Furthermore, MALAT1 promotes EMT and metastasis of lung adenocarcinoma cells in vitro and in vivo. In particular, MALAT1 upregulated the expression of miR-204 target gene SLUG through competitively 'spongeing' miR-204. In summary we unveil a branch of the MALAT1/miR-204/SLUG pathway that regulates the progression of lung adenocarcinoma.

Rate at 120/min provides qualified chest compression during cardiopulmonary resuscitation.

OBJECTIVES: The quality of cardiopulmonary resuscitation (CPR) is a very important prognostic factor for cardiac arrest. Chest compression is thought to be one of the most important aspects of high-quality CPR. Recent studies have prompted that there may be an interaction between chest compression rate and other factors related to the quality of chest compression. We aimed to investigate the effect of different compression rates on chest compression depth, recoil, and rescuers' fatigue point during CPR. METHODS: Participants performed 2 minutes of chest compression-only CPR after the guiding sounds, at 3 rates (100, 120, and 140 compressions/min) in random sequence. A repeated-measures analysis of variance was used to compare the average chest compression depth and other factors related to the quality of chest compression among the groups. RESULTS: As the chest compression rate increases through all the 3 rates, the fractions of chest compressions with complete release and the fractions of chest compressions with sufficient depth were deteriorated at the rate of 140 compressions/min (P < .05), although the average compression depth was above the recommended 2010 guideline depth of 5 cm(P > .05). Of note, the fatigue point at 140 compressions/min happened significantly (P < .05) sooner. CONCLUSION: Our study supported the concern of some that there may be a risk of increasing recommended chest compression rate without providing an upper limit. An appropriate choice may be 120 compressions/min.

[Monitoring the prevalence of schistosomiasis in Erlang Village, Yueyang County from 2005 to 2012].

OBJECTIVE: To understand the dynamics of schistosomiasis situation so as to provide the evidence for formulating the prevention and control scheme in Erlang Village, Yueyang County, Hunan Province. METHODS: The schistosomiasis prevalence and Oncomelania snail status were investigated from 2005 to 2012. RESULTS: The schistosome infection rates of the residents decreased from 2.76% in 2005 to 0.54% in 2012 and the decline rate was 80.43%; the infection rates of the livestock decreased from 8.70% in 2005 to 0 in 2012. No snails were found in the inner embankment over the eight-year period. The density of the infected snails was 0.0005 per 0.1 m(2) in 2005, but no positive snails were found from 2009 to 2012 outside the embankment. CONCLUSION: The schistosome infections of people and livestock and the status of the intermediate host snails have achieved the criteria of schistosomiasis transmission control.

Synthetic peptides containing B- and T-cell epitope of dengue virus-2 E domain III provoked B- and T-cell responses.

Our previous work applied a combination of bioinformatics approaches and in vitro assays to identify the dengue-2 virus (DENV-2)-specific B- and T-cell epitopes. In this report, we first evaluated the antigenicity of both B- and T-cell epitopes reacting with different sera against DENV-2 by ELISA as well as the ability of T-cell epitope to activate CD4(+) T-cell producing IFN-γ using ELISPOT, which showed a specific reactivity between either B- or T-cell epitope and DENV-2 antisera, and a significant increase of IFN-γ producing cells in DENV-2 infected mice. Then, a multi-epitope peptide containing the above B-, T-cell epitopes of envelope domain III (EDIII) of DENV-2 and pan-DR epitope (PADRE) was bioinformatically designed and synthesized. The verification of its immunogenicity and protective effect was performed in in vitro and in vivo experiments. The results showed that a high level of antibody in mice elicited by the multi-epitope peptide was detected by ELISA and the anti-peptide sera binding to the vero cells infected with DEN-2 was observed with immunofluorescence test. More importantly, the peptide could induce lymphoproliferation in vitro and a predominant Th1 type of immune response was examined by flow cytometry. We also found that the virus replication in the mice vaccinated with the multi-epitope peptide was obviously less than that of the control groups. These results may provide some important information for the development of dengue vaccine.