MicroRNA-20a promotes non-small cell lung cancer proliferation by upregulating PD-L1 by targeting PTEN.

Non-small cell lung cancer (NSCLC) remains one of the most common malignant tumors worldwide. The aim of the present study was to investigate the possibility of microRNA-20a (miR-20a) as a biomarker and therapeutic target for the diagnosis and treatment of NSCLC. Bioinformatics prediction, together with functional validation, confirmed miR-20a bound to programmed death ligand-1 (PD-L1) 3'-untranslated region to upregulate PD-L1 expression. Both miR-20a and PD-L1 could promote the proliferation of NSCLC cells. The expression level of PD-L1 was controlled by PTEN; however, further upstream regulation of PD-L1 expression was largely unknown. The present study showed that miR-20a could not restore the inhibition of PD-L1 expression levels by PTEN. Knockdown of PTEN expression upregulated the expression level of PD-L1 and promoted the proliferation of NSCLC cells. PTEN negatively regulated the Wnt/ß-catenin signaling pathway by inhibiting ß-catenin and Cyclin D1. Interestingly, PTEN could reverse miR-20a-mediated proliferation of NSCLC cells and the inhibitory effect was similar to that of XAV-939. miR-20a promotes the proliferation of NSCLC cells by inhibiting the expression level of PTEN and upregulating the expression level of PD-L1. It is suggested that miR-20a could be used as a biomarker and therapeutic target for the treatment of NSCLC.

Target induced framework nucleic acid nanomachine with doxorubicin-spherical nucleic acid tags for electrochemical determination of human telomerase activity.

A stable and enzyme-free method is described for highly sensitive determination of telomerase activity. It is based on the use of a framework nucleic acid (FNA) nanomachine and doxorubicin-spherical nucleic acid (DSNA) tags. Upon incubation with telomerase, the primer-tetrahedron becomes elongated to form the handed swing arm. The extended swing arm autonomously moves along the predefined track consisting of entropy-tetrahedron by consecutive strand displacement under the aid of fuel-tetrahedron. As a result, many (entropy-tetrahedron)-(fuel-tetrahedron) complexes are assembled for combining the DSNA tags. This results in an amplified electrochemical signal, typically measured at around -0.63 V (Ag/AgCl). The use of an enzyme-free FNA nanomachine and of DSNA tags warrants outstandingly high stability and sensitivity. The method shows a broad dynamic correlation of telomerase activity in cell extracts. The analytical range extends from 10 to 1.0 × 104 HeLa cells mL-1 with a lower detection limit of 2 cells mL-1. The differences in telomerase activity between different cancer cells can be easily evaluated. The method was further verified by quantifying telomerase activity of cancer cells in accumulated normal cells. Therefore, the sensing method has great potential for clinical application. Graphical abstractSchematic representation of the electrochemical biosensor based on target induced framework nucleic acid nanomachine with doxorubicin-spherical nucleic acids (DSNA) tags, which can be used to the determination of telomerase activity in accumulated normal cells. dNTP: Deoxynucleotide triphosphates; FT: Fuel-tetrahedron.

Correction: MiR-451 as a new tumor marker for gastric cancer.

[This corrects the article DOI: 10.18632/oncotarget.17239.].

The association between MTHFR polymorphism and cervical cancer.

Cervical cancer is an extremely prevalent disease worldwide. The purpose of this study was to illustrate the relationship between methylenetetrahydrofolate reductase (MTHFR) polymorphisms or methionine synthase reductase (MTRR) polymorphisms and cervical cancer. There were 372 women who performed genetic and folic acid assessments. For the MTHFR C677T, there was no significant difference in the distribution of C allele and T allele in the three groups. However, the mutant C allele of MTHFR A1298C was significantly higher in the cancer group than in the normal group. Similarly, the mutant G allele of MTRR A66G was also higher than the normal group. The serum folic acid levels were gradually decreased with the development of cervical lesions. Serum folate levels in 4-9 ng/ml and ≤4 ng/ml were both significantly associated with cervical cancer risk. However, the MTHFR C677T polymorphism was not associated with the risk of cervical cancer or CIN. In contrast, the MTHFR A1298C polymorphism could increase the risk of both cervical cancer and CIN. In addition, the MTRR A66G polymorphism was only associated with the risk of cervical cancer but not CIN.

MicroRNA-466 with tumor markers for cervical cancer screening.

Cervical cancer is the second most common cancer in women in the world. In this study, we explore tumor markers and microRNA-466 combination for cervical cancer screening. Tumor markers were measured by the methods of electro-chemiluminescent immunoassay and enzyme immunoassay. The microRNA-466 was performed by quantitative real-time polymerase chain reaction. Among normal group, hyperplasia group and cancer group, the CEA expression levels were 2.26 ng/ml, 3.85 ng/ml and 16.08 ng/ml, respectively. While the CA125 expression levels were 13.61 u/ml, 27.32 u/ml and 44.93 u/ml, respectively. The SCCA expression levels were 13.61 ng/ml, 27.32 ng/ml and 44.93 ng/ml, respectively. The expression levels of tumor markers were all gradually increased with the development of cervical lesions. The expression levels of microRNA-466 in cervical cancers (0.62) were greater than that in normal (0.076) and hyperplasia (0.24). The expression of microRNA-466 was correlated with lymphnode metastasis (P=0.000). There is a lower overall survival rate of patient with large tumor or lymphnode metastasis. Thus, the combination of tumor markers and microRNA-466 can be useful for early detection of cervical cancer and indicators for advanced stage and prognosis of the disease.

MiR-451 as a new tumor marker for gastric cancer.

Gastric cancer is the second most common malignancy in China. However, the prognosis for gastric cancer patients remains poor. The purpose of this study was to investigate whether miR-451 was a potential prognostic biomarker for gastric cancer. Fresh tissues were immediately frozen in liquid nitrogen until use. The plasma was extracted and quantitative real-time polymerase chain reaction was performed to detect miR-451 expression. The Student's t test analysis and multivariate Cox regression analysis were performed to analyze expression of miR-451. The analysis results showed that the expression level of miR-451 was decreased expression in gastric cancer tissue. The down-regulation of miR-451 tended to be positively correlated with tumor stage, lymphatic metastasis and shorter overall survival of patients. MiR-451 may be a potential biomarker and a potential therapeutic target for the diagnoses and prognosis of gastric cancer.

A New MicroRNA Expression Signature for Cervical Cancer.

BACKGROUND: Cervical cancer is the second most common cancer among women worldwide. The potential of microRNAs as novel biomarkers in cervical cancer is growing. OBJECTIVES: In this study, we investigated the functions and targets of miR-466 in cervical cancer tissues. METHODS: Fresh cervical tissues were obtained from 157 patients with cervical cancer, cervical intraepithelial neoplasia (CIN), and healthy controls, and the tissues were immediately frozen in liquid nitrogen until use. The RNA was extracted and quantitative real-time polymerase chain reaction (PCR) was performed. RESULTS: A total of 157 participants were summarized, including 56 patients with cervical cancer, 60 patients with CIN, and 49 healthy controls. The expression levels of miR-466 in cervical cancers (0.68) were higher than that in healthy controls (0.082) (P < 0.01). The average fold changes of miR-466 in the patients with CIN group and people group were 0.28 and 0.082, respectively (P < 0.01). It was a statistically significant difference in patients with lymph node involvement (P = 0.022). However, the expression of miR-466 was not correlated with International Federation of Gynecology and Obstetrics stages, tumor size, or vascular invasion (P = 0.506, P = 0.667, and P = 0.108, respectively). CONCLUSIONS: Our results indicate that the aberrant expression of miR-466 is closely associated with the occurrence and development of cervical cancer.

Detection of New Delhi Metallo-Beta-Lactamase (Encoded by blaNDM-1 ) in Enterobacter aerogenes in China.

BACKGROUND: The increase in blaNDM-1 in Enterobacteriaceae has become a major concern worldwide. In previous study, we investigated clonal dissemination and mechanisms of resistance to carbapenem in China. METHODS: We carried out retrospective surveillance for blaNDM-1 among carbapenem-resistant enterobacter strains, which were isolated from patients at our hospital by bacterial strains selection, antimicrobial susceptibility testing, species identification, and molecular detection of resistance gene. RESULTS: We found three blaNDM-1 -positive isolates which were identified as Enterobacter aerogenes in clinical patients in China. The blaNDM-1 -positive Enterobacter aerogenes isolates were first found. CONCLUSION: It is important to mandate prudent usage of antibiotics and implement infection control measures to control the spread of these resistant blaNDM-1 -positive strains.

Reference Intervals of Routine Coagulation Assays During the Pregnancy and Puerperium Period.

BACKGROUND: Significant changes occur in the coagulation and fibrinolytic systems during pregnancy and puerperium in the plasma levels. However, reference ranges based on healthy people are not optimal for informing clinical decisions during the pregnancy and puerperium. Therefore, it is essential to explore coagulation assays' reference ranges during the pregnancy and puerperium. METHODS: Prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), fibrinogen (Fib.), and D-dimer were all measured according to the manufacturer's specifications and laboratory standard operating procedure of the STA-R evolution coagulation analyzer. A total of 11,601 women were enrolled in this study. RESULTS: The reference ranges for PT, APTT, TT, Fib., and D-dimer in nonpregnancy period were 10.87-13.76 s, 29.22-44.61 s, 15.39-20.15 s, 1.59-3.97 g/l, and 0-0.56 mg/l, respectively. In early-pregnancy period, the ranges were 11.14-14.07 s, 29.97-44.69 s, 14.92-19.03 s, 1.98-4.13 g/l, and 0-1.67 mg/l, respectively. In midpregnancy period, the ranges were 9.98-12.84 s, 28.53-40.70 s, 13.51-19.82 s, 2.63-5.19 g/l, and 0-2.81 mg/l, respectively. In late-pregnancy period, the ranges were 9.48-12.58 s, 28.61-40.80 s, 14.10-19.61 s, 2.80-5.56 g/l, and 0-27.08 mg/l, respectively. In puerperium period, the ranges were 10.85-13.72 s, 30.51-43.02 s, 15.31-19.64 s, 1.14-5.07 g/l, and 1.27-4.85 mg/l, respectively. CONCLUSION: We presented reference intervals for coagulation assays from the nonpregnancy to puerperium period that can be adopted in other laboratories after further validation.

Newborn screening for congenital hypothyroidism in Henan province, China.

BACKGROUND: Congenital hypothyroidism is the most common congenital endocrine disorder. The study aimed to determine the congenital hypothyroidism incidence by newborn screening programs in Henan Province, China. METHODS: The screening programs for congenital hypothyroidism are based on the measurement of TSH in dried blood spots. The TSH concentration was measured in the dry blood spot specimen using a DELFIA fluoroimmunoassay. The TSH cutoff concentration was 8mU/l. RESULTS: The total coverage and the incidence of congenital hypothyroidism were 24.85% (5,142,148/20,694,441) and 0.37‰ (1992/5,142,148), respectively. The coverage and the incidence of CH were only 0.58% (4526/784,580) and 0.22‰ (1/4526) in 1997, respectively. However, the coverage and the incidence of CH were increased to 74.67% (1,203,278/1,611,582) and 0.32‰ (389/1,203,278). There were no significant differences in the number of congenital hypothyroidism between males and females (P>0.05). The number of congenital hypothyroidism was increased year after year. CONCLUSIONS: The newborn screening program for CH is successful and quite effective.

Performance of the HPV-16 L1 methylation assay and HPV E6/E7 mRNA test for the detection of squamous intraepithelial lesions in cervical cytological samples.

HPV-16 L1 methylation and E6/E7 mRNA have suggested that they had close relationship with cervical neoplastic progression. This study aimed to evaluate the clinical performance of the HPV-16 L1 methylation assay and E6/E7 mRNA test for detecting high-grade cervical lesions (CIN2+). A total of 81 women with liquid-based cytology (LBC) samples, histological results, and positive HPV-DNA test for HPV type 16 only were included in this study. HPV-16 L1 methylation and E6/E7 mRNA levels were measured using methylation-sensitive high resolution melting (MS-HRM) analysis and Quantivirus®HPV E6/E7 RNA 3.0 assay (bDNA), respectively, in the same residue of LBC samples. The current date showed a positive correlation between the HPV-16 L1 methylation and the E6/E7 mRNA levels. The L1 methylation and mRNA levels both increased with disease severity. The mRNA test method showed higher sensitivity and NPV (98.0 and 91.7% vs. 89.8 and 80.8%), while lower specificity and PPV (34.4 and 69.6% vs. 65.6 and 80.0%), than the L1 methylation assay for detecting histology-confirmed CIN2+. When using the detection method of mRNA test combined with L1 methylation assay, we obtained a sensitivity of 89.8% and a specificity of 71.9%. These findings suggest that assessment of HPV-16 L1 methylation testing combined with E6/E7 mRNA testing may be a promising method for the triage of women with HPV type 16 only.

High-resolution melting analysis of HPV-16L1 gene methylation: A promising method for prognosing cervical cancer.

OBJECTIVE: Methylation-sensitive high-resolution melting (MS-HRM) is a new technique for DNA methylation analysis, but it is rarely used for the detection of viral DNA methylation. In this study, we investigated the HPV-16L1 gene methylation that is detected by MS-HRM as a potential biomarker for prognosing cervical dysplasia and cancer. DESIGN AND METHODS: A total of 114 HPV-16 infected patients (normal (17), CIN1 (25), CIN2 (29), CIN3 (32), SCC (11)) who underwent liquid-based cytology test and biopsy were included in this study. 17 cases with HPV-16 infection and negative cytologic and histologic results served as the control group. The HPV-16L1 gene methylation statuses of these samples were investigated using a methylation-sensitive high-resolution melting (MS-HRM) assay after bisulfite modification. RESULTS: The HPV-16L1 gene methylation statuses of all the 114 specimens were successfully detected by MS-HRM, and we observed increasing methylation levels in severe lesions, as determined using histologic assays. In addition, the methylation levels of CIN2+ (CIN2, CIN3 and SCC) were significantly higher than that of CIN2- (normal and CIN1, P<0.001). When taking CIN2+ as the reference, our HPV-16L1 DNA methylation assay achieved 91.7% sensitivity and 59.5% specificity, respectively. CONCLUSIONS: The results of the present work demonstrated that HPV-16L1 gene methylation was closely associated with cervical precancerosis and cancer. Moreover, using MS-HRM to detect HPV-16L1 gene methylation may be a powerful assay for the triage of HPV-16-positive females, which could identify patients with high risk of invasive cancer.

Epidemiology and genotype distribution of human papillomavirus (HPV) in women of Henan Province, China.

BACKGROUND: Human papillomavirus (HPV) infection is the commonest sexually transmitted infection, which is associated with various clinical conditions. This study aimed to determine the distribution of HPV genotypes in the women of Henan Province, China. METHODS: Cervical samples were collected by liquid-based method and consecutively evaluated cervical cytology and the presence of HPV DNA. Cytological classification was made according to the Bethesda 2001 criteria. HPV DNA was tested with xMAP technology by Luminex200™. RESULTS: In cervical abnormalities, the infection rate of HPV was 84.0%, single type was 71.0%, multiple type was 13.0%, high risk HPV was 78.0% and low risk HPV was 8.0%. The most common genotypes found were HPV16, 52, 58, 33, 18, 6 and 39. The most common HPV genotypes were HPV16, 52, 6, 58 and 33 in NILM, HPV16, 52, 18, 58 and 6 in ASCUS, HPV52, 16, 58, 6 and 39 in LSIL, HPV16, 33, 58, 18 and 51 in HSIL, and HPV16, 18, 33, 58 and 52 in ICC, respectively. The prevalence of single HPV and multiple HPV was 64.8% and 13.3%, respectively. Age-specific prevalence of multiple HPV exhibited a "U" shaped curve. CONCLUSIONS: Single HPV genotype infection was predominantly detected in different groups of cervical lesions in Henan Province, and HPV16, 52, 58, 33, 18 and 6 were the priority HPV types.

Quantivirus® HPV E6/E7 RNA 3.0 assay (bDNA) is as sensitive, but less specific than Hybrid Capture 2 test.

Human papillomavirus (HPV) infection is the primary cause of cervical cancer. The Quantivirus(®) HPV E6/E7 RNA 3.0 assay (DiaCarta, CA, USA) detects E6/E7 mRNA of 13 high risk subtypes and 6 low risk subtypes. Cervical specimens collected in PreservCyt were processed for HPV detection. Cervical biopsies were taken only from those women with abnormal colposcopy. 200 out of 272 (73.5%) cases were mRNA positive. The percentage of HPV E6/E7 mRNA positive samples increases with the severity of the cytological diagnosis, but not in histological diagnosis. In 146 patients with both tests, the E6/E7 mRNA assay had significant higher positivity rate than the Hybrid Capture 2 assay (75.3% versus 62.3%). The HPV mRNA assay and the HC2 assay had the same sensitivity of high grade cervical intraepithelial neoplasia (CIN 2+), 82.4% (14/17) (95% confidence interval [CI], 64.3, 100). However, the specificity of CIN 2+ for the HPV mRNA assay was significantly lower than HC2 assay. Receiver operating characteristic curve analysis was used to compare the diagnostic performance of the E6/E7 mRNA and HC2. E6/E7 mRNA achieved 58.8% sensitivity with 74.1% specificity, HC2, achieved 47.1% sensitivity with 70.7% specificity. The overall performance of HPV E6/E7 mRNA assay for detecting CIN 2+ was lower than HC2. This study does not support the use of this assay in screening for cervical cancer prevention alone.

New technology for cervical cancer screening.

BACKGROUND: Cervical cancer is the second most common cancer among women worldwide. With the introduction of organized cervical cytological screening programs, the incidence of cervical cancer has been dramatically reduced. OBJECTIVES: This study aimed to determine the new technology that can potentially afford unique advantages for cervical cancer screening. METHODS: Cervical specimens collected in PreservCyt were processed for ThinPrep cytological test, the new technology test and human papillomavirus detection. RESULTS: The concordance between the new technology and ThinPrep cytological test was 96.34%, with 931 cases positive and 148 cases negative with both tests (κ = 0.857). The sensitivity and the specificity of the new technology were 99.04% (931/940) and 82.22% (148/180), respectively. Youden index was 0.81. The positive predictive value and the negative predictive value were 96.68% (931/963) and 94.27% (148/157), respectively. In the 124 positive cases of the new technology, human papillomavirus DNA test was positive in 109 cases (87.9%) and negative in 15 cases (12.1%). Compared to the histopathological diagnosis, the sensitivity and the negative predictive value of the new technology were 98.57% (69/70) and 95.45% (21/22), respectively. CONCLUSIONS: The screening design will enable evaluation of several competing screening technologies in reducing the incidence of and mortality from cervical cancer. In particular, if the new technology is used as the screening test, it can be a quick screening test and does not depend on the subjective judgment of the doctors. As such, it could potentially afford unique advantages for screening.