Expression and Clinical Values of Serum miR-155 and miR-224 in Chinese Patients with HCV Infection.

BACKGROUND: To investigate the expression of serum miR-155 and miR-224 among patients with hepatitis C virus (HCV) infection and analyze their clinical values. METHODS: A total of 116 patients suffering from HCV infection admitted to our hospital and 70 healthy subjects were selected. According to the diagnostic results, patients with HCV infection were divided into 48 cases of chronic hepatitis C (CHC), 43 cases of liver cirrhosis and 25 cases of hepatocellular carcinoma (HCC). The expression signature for miR-155 and miR-224 was detected in serum samples. ROC curve and Pearson correlation test were conducted to investigate their diagnostic value and correlation. RESULTS: The expression extent for serum miR-155 and miR-224 increased along with the increase of malignancy (all P < 0.05). According to ROC curve, the area under the curve (0.918, 95% CI: 0.856-0.974) of miR-155 and miR-224 combined in the diagnosis of HCC was the largest, and its sensitivity and specificity were 93.0% and 86.2%. There is a positive relationship for expression level between miR-155 and miR-224 in CHC and HCC group (all P < 0.001). CONCLUSION: miR-155 and miR-224 are remarkably increased in patients suffering from HCV infection. The combination of miR-155 and miR-224 has a good diagnostic value for HCC caused by HCV infection.

Automatic Acetowhite Lesion Segmentation via Specular Reflection Removal and Deep Attention Network.

Automatic acetowhite lesion segmentation in colposcopy images (cervigrams) is essential in assisting gynecologists for the diagnosis of cervical intraepithelial neoplasia grades and cervical cancer. It can also help gynecologists determine the correct lesion areas for further pathological examination. Existing computer-aided diagnosis algorithms show poor segmentation performance because of specular reflections, insufficient training data and the inability to focus on semantically meaningful lesion parts. In this paper, a novel computer-aided diagnosis algorithm is proposed to segment acetowhite lesions in cervigrams automatically. To reduce the interference of specularities on segmentation performance, a specular reflection removal mechanism is presented to detect and inpaint these areas with precision. Moreover, we design a cervigram image classification network to classify pathology results and generate lesion attention maps, which are subsequently leveraged to guide a more accurate lesion segmentation task by the proposed lesion-aware convolutional neural network. We conducted comprehensive experiments to evaluate the proposed approaches on 3045 clinical cervigrams. Our results show that our method outperforms state-of-the-art approaches and achieves better Dice similarity coefficient and Hausdorff Distance values in acetowhite legion segmentation.

MicroRNA-19a mediates neuroprotection through the PTEN/AKT pathway in SK-N-SH cells after oxygen-glucose deprivation/reoxygenation injury.

Ischemic stroke is one of the most common public health problems worldwide. The aim of the present study was to investigate the role of miR-19a and its possible target genes in SK-N-SH cells subjected to oxygen-glucose deprivation/re-oxygenation (OGD/R) injury. SK-N-SH cells are a suitable model for host transfection. SK-N-SH cells were transfected with miR-19a mimic or inhibitor and PTEN-small interfering (si) RNA in order to alter the expression of miR-19a, PTEN and AKT. The expression changes in acute cerebral ischemic injury (ACII) were verified using RT-qPCR and Western blotting. Expression changes and the association between miR-19a and PTEN following OGD/R were also assessed using a double luciferase analysis. In addition, cell viability and apoptosis were measured using an MTT and flow cytometry. miR-19a was downregulated; however, PTEN was markedly increased following OGD/R injury. miR-19a mimics increased cell viability, decreased cell apoptosis of SK-N-SH cells following OGD/R, which effects was similar to PTEN siRNA; however, miR-19a inhibitor had the opposite roles with miR-19a mimics. The present study provides novel information about the cell apoptosis and invasion mechanisms associated with the miR-19a/PTEN/AKT pathway and may present a potential therapeutic approach for OGD/R injury.

Association between TLR2 Arg677Trp polymorphism and tuberculosis susceptibility: A meta-analysis.

BACKGROUND: Toll-like receptor 2 (TLR2) is an important member of TLRs, which is significant in the initial of inflammatory response against bacteria. Several studies have been conducted to investigate the TLR2 Arg677Trp polymorphism and Tuberculosis (TB) susceptibility. Unfortunately, the results of previous studies were inconsistent. OBJECTIVES: The aim of present study was to investigate the relationship between TLR2 Arg677Trp polymorphisms and TB susceptibility. METHODS: The association between the TLR2 Arg677Trp polymorphism and TB susceptibility was assessed by odds ratios (OR) together with their 95% confidence intervals (95%CI). RESULTS: Six case-control studies were enrolled in the meta-analysis. Overall, significant association between TLR2 Arg677Trp polymorphism and TB risk were found neither under allele contrast (C vs. T: OR = 0.59, 95%CI = 0.28-1.23, P = 0.158) nor under recessive genetic model (CC vs. CT/TT: OR = 0.43, 95%CI = 0.12-1.51, P = 0.188). CONCLUSION: We conclude that TLR2 Arg677Trp polymorphism is not associated with TB susceptibility.

Association between the IL-10 and IL-6 polymorphisms and brucellosis susceptibility: a meta-analysis.

BACKGROUND: Brucellosis is a quite normal zoonotic infection, which is caused by immediate contact with animals infected with Brucella or its products. IL-10 (- 1082 G/A, - 819 C/T, - 592C/A) and IL-6 -174 G/C polymorphisms have a great relationship with IL-10 and IL-6 production, which brings about Brucellosis pathogenesis and development. So far, the results of published literatures were controversial. Now, we perform a meta-analysis in different ethnic populations to get a more precise estimate of above polymorphisms with Brucellosis susceptibility. METHODS: Both OR and corresponding 95%CI were enrolled to make an assessment of the association strength through extracting genotyping frequency of cases and controls. The χ2-test based Q-statistic and I2 statistics were applied. If there was no evident heterogeneity, the fixed-effects model would be applied. If not, the random-effects model would be used. RESULTS: The significant associations were only found in Asian population of - 819 loci under three genetic models as follows: (Allele model: OR = 0.60, 95%CI = 0.44-0.82, P = 0.001), (homozygote comparison: OR = 0.24, 95%CI = 0.09-0.62, P = 0.003), (recessive genetic model: OR = 0.22, 95%CI = 0.05-0.91, P = 0.036). CONCLUSION: In conclusion, IL-10 - 819 loci polymorphism contributes no risk to Caucasian population but may be associated with decreased risk in Asian population. And IL-10 -1082 G/A, 592 loci and IL-6 -174 G/C polymorphism are not associated with Brucellosis risk.

Automatic CIN Grades Prediction of Sequential Cervigram Image Using LSTM With Multistate CNN Features.

Cervical cancer ranks as the second most common cancer in women worldwide. In clinical practice, colposcopy is an indispensable part of screening for cervical intraepithelial neoplasia (CIN) grades and cervical cancer but exhibits high misdiagnosis rate. Existing computer-assisted algorithms for analyzing cervigram images have neglected that colposcopy is a sequential and multistate process, which is unsuitable for clinical applications. In this work, we construct a cervigram-based recurrent convolutional neural network (C-RCNN) to classify different CIN grades and cervical cancer. Convolutional neural networks are leveraged to extract spatial features. We develop a sequence-encoding module to encode discriminative temporal features and a multistate-aware convolutional layer to integrate features from different states of cervigram images. To train and evaluate the performance of C-RCNN, we leveraged a dataset of 4,753 real cervigrams and obtained 96.13% test accuracy with a specificity and sensitivity of 98.22% and 95.09%, respectively. Areas under each receiver operating characteristic curves are above 0.94, proving that visual representations and sequential dynamics can be jointly and effectively optimized in the training phase. Comparative analysis demonstrated the effectiveness of the proposed C-RCNN against competing methods, showing significant improvement over only focusing on a single frame. This architecture can be extended to other applications in medical image analysis.

Association between the IL-10-1082G/A, IL-10-819T/C and IL-10-592A/C polymorphisms and Brucellosis susceptibility: a meta-analysis.

Brucellosis is a widespread zoonosis caused by small bacteria of the genus Brucella. The promoter polymorphisms of IL-10 (-1082 loci, -819 loci and -590 loci) are closely related to the production of IL-10, leading to the alteration of development and pathogenesis of Brucellosis. However, the previous results were controversial. In the present study, we conduct the meta-analysis to get a more precise result of IL-10 polymorphisms with Brucellosis risk. The quality of the studies was assessed according to a predefined scale. The odds ratio (OR) and 95% confidence interval (CI) were counted to evaluate the association strength. No significant association was found between position -1082 loci or -590 loci polymorphism and Brucellosis risk. The significant association was found in Asian population of position -819 (T vs. C: OR 0.60, 95% CI 0.44-0.82, P = 0.001), homozygote comparison (TT vs. CC: OR 0.24, 95% CI 0.09-0.62, P = 0.003) and recessive genetic model (TT vs. TC/CC: OR 0.22, 95% CI 0.05-0.91, P = 0.036). The present meta-analysis demonstrates that IL-10-819 loci polymorphism is not associated with Brucellosis risk of Caucasian population but may contribute a decreased risk to Asian population. And neither IL-10-1082 loci nor -592 loci polymorphism is associated with Brucellosis risk.

Quantitative assessment of the association between IL-10 -592 A/C polymorphism and Kawasaki disease risk in Chinese population: evidence from a meta-analysis.

BACKGROUND: IL-10, as a proinflammatory and anti-inflammatory cytokine, has been thought to have an important role in the development of Kawasaki disease. Variation in the IL-10 gene might lead to altered protein production, which may result in Kawasaki disease. Several studies have been performed to investigate the IL-10 -592 A/C polymorphism and Kawasaki disease risk. Unfortunately, the results of previous studies were inconsistent. Therefore, we performed a meta-analysis to derive a more precise estimation of the association between the IL-10 -592 A/C polymorphism and Kawasaki disease risk. METHOD: The association between the IL-10 -592 A/C polymorphism and Kawasaki disease risk was assessed by odds ratios (ORs) together with their 95% confidence intervals (CIs). Six studies were enrolled in the present meta-analysis. RESULTS: Overall, no significant association between IL-10 -592 A/C polymorphism and Kawasaki disease risk was found under allele contrast (A versus C: OR=0.95, 95% CI=0.77-1.18, p=0.668), homozygote comparison (AA versus CC: OR=0.86, 95% CI=0.56-1.31, p=0.475), heterozygote comparison (CA versus CC: OR=0.88, 95% CI=0.65-1.19, p=0.479), recessive genetic model (AA versus CA/CC: OR=0.96, 95% CI=0.73-1.28, p=0.801), or dominant genetic model (AA/CA versus CC: OR=0.85, 95% CI=0.64-1.13, p=0.275). CONCLUSIONS: We conclude that IL-10 -592 A/C polymorphism was not associated with Kawasaki disease risk in the Chinese population. However, more primary large-scale and well-designed studies are still required to further evaluate the interaction of IL-10 -592 A/C polymorphism with Kawasaki disease risk.

Association between TLR2 + 2477G/A polymorphism and bacterial meningitis: a meta-analysis.

Toll-like receptor 2 (TLR2) is a key member of TLRs, which is crucial in the initial inflammatory response against bacteria. TLR2, is also the initial barrier against bacterial infection and plays an important role in recognising a variety of bacterial lipoproteins. Several studies have been performed to investigate the TLR2 + 2477G/A polymorphism and bacterial meningitis susceptibility. Unfortunately, the results of previous studies were controversial. Therefore, we performed a meta-analysis to derive a more precise estimation of the association. The association between the TLR2 + 2477G/A polymorphism and bacterial meningitis susceptibility was assessed by odds ratios together with their 95% confidence intervals (CI). Six studies were enrolled in the present meta-analysis. Overall, no significant association between TLR2 + 2477G/A polymorphism and bacterial meningitis risk were found under allele contrast (A vs. G: OR = 1.15, 95% CI = 0.93-1.43, P = 0.202), recessive genetic model (AA vs. AG/GG: OR = 1.12, 95% CI = 0.90-1.41, P = 0.313). The significant association was found between TLR2 + 2477G/A polymorphism and pneumococcal meningitis risk under allele contrast (A vs. G: OR = 1.54, 95% CI = 1.01-2.36, P = 0.046), recessive genetic model (AA vs. AG/GG: OR = 1.63, 95% CI = 1.03-2.57, P = 0.035). We conclude that TLR2 + 2477G/A polymorphism is not associated with meningococcal meningitis risk but contributes an increased risk of pneumococcal meningitis.

Association of GPIa and COX-2 gene polymorphism with aspirin resistance.

OBJECTIVE: This study aimed to explore the association between GPIa, COX-2 gene polymorphisms and aspirin resistance in the ischemic stroke patients from the southern part of Jiangsu province. METHODS: In all, 97 patients with acute ischemic stroke were enrolled in the study. GPIa gene polymorphism at 807C>T (rsl126643) locus and COX-2 gene polymorphism at -765G>C (rs20417) locus were genotyped by PCR pyrosequencing technology. Patients were divided into the aspirin sensitivity (AS) group and aspirin resistance (AR) group according to the platelet aggregation rate. The relationship between the two gene polymorphisms and aspirin resistance was investigated and analyzed. RESULTS: The distribution of the genotype (CC, CT, TT, CT + TT, and CC) and the frequency of allele T of GPIa gene at 807C>T locus were significantly different in AS and AR groups in female patients (P < .05). Logistic regression analysis showed that the genotype of CT+TT at 807C>T locus was significantly correlated with AR after adjustment for relative factors (P = .047, OR = 4.856, 95% CI: 1.020-23.108). There were no significant differences in the genotype distribution and allele frequency of the COX-2 gene -765G>C site between two groups (P > .05). CONCLUSION: GPIa gene polymorphism at 807C>T locus was associated with AR in Chinese Han females, and the expression of allele T increased the incidence of AR. The gene polymorphism of COX-2 gene at -765G>C locus was not significantly correlated with AR.

High-resolution melting analysis for rapid and sensitive NOTCH1 screening in chronic lymphocytic leukemia.

Chronic lymphocytic leukemia (CLL) is a biological and clinical heterogeneous disease. Activating mutations of NOTCH1 have been implicated to be associated with adverse prognosis in CLL. The objective of the present study was to develop an effective high-resolution melting (HRM) assay for detecting NOTCH1 mutations. Genomic DNA (gDNA) extracted from 133 CLL patients was screened by HRM assay, and the results were compared with the data obtained using direct sequencing. The relative sensitivity of the HRM assay and direct sequencing was evaluated using diluted gDNA with different NOTCH1 mutational frequencies. The HRM assay was able to detect and discriminate samples with NOTCH1 mutations from the wild-type template in CLL. Eight of the 133 CLL patients (6.02%) were scored positively for NOTCH1 mutations in the HRM assay. The results of the NOTCH1 mutations detected by HRM analysis achieved 100% concordance with those determined from direct sequencing. HRM had a higher sensitivity (1%) and shorter turn-around time (TAT), compared to direct sequencing. In conclusion, the HRM assay developed by us was confirmed to be a rapid, sensitive, and promising approach for high-throughput prognostic NOTCH1 screening in CLL. It enables real-time NOTCH1 evaluation, which is of great significance in clinical practice and may facilitate the decision-making of clinicians in CLL.

The NF-κB subunit RelB regulates the migration and invasion abilities and the radio-sensitivity of prostate cancer cells.

NF-κB subunits play important roles in carcinogenesis of a variety of human malignancies and response to cancer therapy; however, the contribution of an individual subunit has not been thoroughly defined. Constitutive activation of the canonical NF-κB subunit is a critical event in prostate carcinogenesis. Recent findings point out that RelB, which contributes to the non-canonical NF-κB activity, functions importantly in the prostate cancer progression. Here, we investigated systemically the functional roles of RelB in prostate cancer and examine its significance as a therapeutic target. Targeting RelB using short hairpin RNA approach in androgen-independent DU145 prostate cancer cells interfered with various biological behaviors of cells. We observed that RelB knockdown inhibited prostate cancer cell growth, migration, and invasion, and enhanced proteasome inhibitor sensitivity. The altered expression of anti-apoptotic gene Bcl-2 played critical roles in regulating both spontaneous and radiation-induced apoptosis in the presence of RelB knockdown. For the first time, we showed that RelB knockdown significantly attenuated the migration and invasion of DU145 prostate cancer cells, due to the reduction of integrin ß-1. Collectively, we provided evidence that RelB functioned as an oncogene in prostate cancer. Developing a RelB-targeted therapeutic intervention, is valuable in treating advanced, metastatic prostate cancer.

[Evaluation of a new method and instrument for detection platelet aggregation function and its clinical application].

This study was purpose to evaluate a new method and instrument for detecting platelet aggregation function, establish the reference intervals for PL-11 platelet analyzer, and evaluate its clinical application. The evaluation was based on the guidelines of Clinical and Laboratory Standards Institute (CLSI or NCCLS) and Clinical Laboratory Improvement Amendment 88. Intravenous blood samples anticoagulated with sodium citrate were detected by PL-11 platelet analyzer. The reference intervals were defined after statistic analysis. The clinical diagnostic significance of the PL-11 platelet analyzer was evaluated by testing the change rate of platelet maximum aggregation rate (MAR) of acute cerebral infarction (ACI) patients in the department of Neurology who took clopidogrel 7 d before and after. The result showed that all the parameters meet the standard of CLIA'88. The platelet MAR of 247 healthy volunteers which was induced by PLR-06, PLR-07, PLR-09 and PLR-10, was detected by the PL-11 platelet analyzer, respectively. The MAR is 58.8 ± 10.1 (%), 61.2 ± 11.8 (%), 51 ± 10.2 (%), 53.1 ± 9.2 (%), respectively. The MAR of ACI patients is significantly lower than that after taking clopidogrel. It is concluded that the PL-11 platelet analyzer is an ideal platelet function detector for early warning and diagnosis of thromboembolic disease, which is worthy to be extended and applied.

Inhibition of MCF-7 breast cancer cell-induced platelet aggregation using a combination of antiplatelet drugs.

Cancer metastasis is a highly coordinated and dynamic multistep process in which cancer cells interact with a variety of host cells. Morphological studies have documented the association of circulating tumor cells with host platelets. Tumor cell-induced platelet aggregation (TCIPA) contributes significantly to hematogenous metastasis; however, the molecular mechanisms involved in breast cancer TCIPA are poorly characterized. In this study, MCF-7 metastatic human breast cancer cells induced dose-dependent aggregation of washed platelets. Four major platelet activation pathways, glycoprotein (GP)-Ib-IX, GPIIb/IIIa, thromboxane (TX)-A2 and adenosine diphosphate (ADP) were activated during TCIPA and were inhibited by their respective inhibitors, 7E3, SZ-1, aspirin and apyrase. Pretreatment of platelets with 7E3, SZ-1 or apyrase significantly inhibited TCIPA, while pretreatment with aspirin had no effect. Moreover, combined pretreatment of platelets with 7E3, SZ-1 and apyrase significantly inhibited TCIPA, compared to single inhibitors. Combinations of antiplatelet drugs may represent a promising strategy to prevent cancer metastasis.

A high-throughput biotin-avidin-ELISA for studying expression of platelet membrane glycoproteins and its clinical application.

Platelet membrane glycoproteins (GPs) are critical for normal platelet adhesion, activation and aggregation. To define the abnormalities in surface GP expression on circulating platelets and provide a better biomarker of bleeding and thrombotic disorders, we have developed a accurate, time-saving and high-throughput biotin-avidin enzyme-linked immunosorbent assay (BA-ELISA) with the monoclonal antibodies (mAbs), 7E3 against the complex of GPIIb and GPIIIa (GPIIb/IIIa), SZ-51 against P-selectin, and SZ-2 against GPIb, respectively. The levels of P-selectin and GPIIb/IIIa were measured in patients with acute myocardial infarction (AMI), intracerebral hemorrhage (ICH), or diabetes mellitus (DM)) and healthy subjects. Inhibition of GP expression was evaluated with SZ-21, an inhibitory mAb against GPIIIa and aspirin, respectively. The sensitivity of BA-ELISA is high enough to detect platelet count as low as 3.13 x 10(9)/L in platelet-rich plasma (PRP). Both the inter-assay and intra-assay coefficient variation are less than 10%. Adenosine diphosphate (ADP)-induced or non-ADP-induced expression of P-selectin and GPIIb/IIIa was significantly higher in AMI, ICH or DM than that in controls (P < 0.01 for each). Either SZ-21 or aspirin can inhibit the ADP-induced expression of P-selectin and GPIIb/IIIa. Importantly, a high correlation was detected between BA-ELISA and flow cytometry methods. These observations indicate that BA-ELISA is a sensitive and high-throughput assay for evaluating platelet GP expression. The newly developed BA-ELISA can be popularized in community hospitals, because it does not require sophisticated equipments and reagents. This method is suitable for screening inhibitors of platelet activation and has a potential in use for diagnostic purposes.