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Stress might exaggerate the compulsion and impair the working memory of patients with obsessive-compulsive disorder (OCD). This study evaluated the effect of stress on the cognitive neural processing of working memory in OCD and its clinical significance using a “number calculation working memory” task. Thirty-eight patients and 55 gender- and education-matched healthy controls were examined. Stress impaired the performance of the manipulation task in patients. Healthy controls showed less engagement of the medial prefrontal cortex and striatum during the task under stress versus less stress, which was absent in the patients with OCD. The diagnosis × stress interaction effect was significant in the right fusiform, supplementary motor area, precentral cortex and caudate. The failure of suppression of the medial prefrontal cortex and striatum and stress-related hyperactivation in the right fusiform, supplementary motor area, precentral cortex, and caudate might be an OCD-related psychopathological and neural response to stress.
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To detect the levels of miR-146a and miR-155 in different samples from chronic hepatitis B (CHB), reveal whether there is a correlation between the 2 miRNAs in different samples, and to provide a theoretical basis for sample choice of miRNA research in liver. Methods: Real-time PCR was conducted to examine the expression of miR-146a and miR-155 in the plasma, peripheral blood mononuclear cell (PBMC), and liver tissues from 41 CHB patients who underwent nucleoside analogues antiviral therapy for 104 weeks. Correlations between the levels of miR-146a and miR-155 among the 3 samples were analyzed. Results: The expressions of miR-146a and miR-155 in the plasma, PBMC and liver tissues were significantly down-regulated at the 104th week than those at the baseline (all P0.05). Conclusion: Compared with PBMC, miR-146a and miR-155 from plasma can better reflect the expression in the liver tissues, suggesting that plasma can be applied in the mechanism research on miR-146a and miR-155 in the liver diseases instead of liver tissues.
Subject(s)
Humans , Hepatitis B, Chronic , Genetics , Leukocytes, Mononuclear , MicroRNAs , Genetics , Real-Time Polymerase Chain ReactionABSTRACT
Objective:To investigate expression profiles of the plasma exosomal miRNAs of the chronic hepatitis B (CHB) patients with persistently normal alamine aminotransferase (PNALT) for the first time and try to find exosomal miRNAs which could reflect liver inflammation better.Methods:Five CHB patients with liver tissue inflammation grade ≥A2 of PNALT and 5 CHB patients with liver tissue inflammation grade ＜A2 of PNALT were enrolled and their blood samples were collected.The exosomes were extracted from these blood samples and measured by electron microscope to determine the extraction effect.The exosomal miRNAs were extracted and sent for high throughput sequencing,and the expression of exosomal miRNAs in the 2 groups of patients was analyzed.Results:Under the electron microscope,exosomes were small membranous vesicles with 30-100 nm in diameter.The peak value of particle size ranged from 10 to 100 nm.High throughput sequencing showed that there were 591 differentially expressed exosomal miRNAs between the 2 groups.Compared with the control group,18 exosomal miRNAs were up-regulated and 6 exosomal miRNAs were down-regulated in PNALT patients with the liver tissue inflammation grade ≥ A2.Conclusion:Exosomal miRNAs in the CHB patients with PNALT who have the different grades of liver inflammation are differently expressed.Some of the differently expressed exosomal miRNAs are expected to be sensitive biomarkers for timely assessment of liver inflammation in the CHB patients with PNALT.
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Objective To evaluate the accuracy and feasibility of time-resolved immunofluorometric assay (TRI FA) for detection of HBsAg based on Abbott automated chemiluminescence immunoassay(CMIA),so as to carry out this project in primary hospitals,and provide reference for individual antiviral strategy and prediction of therapeutic effect.Methods Serum of 157 patients infected with hepatitis B virus were detected with CMIA and TRIFA,specimens with HBsAg titers exceeding the detection limit were firstly diluted,then performed quantitative analysis.HBsAg levels were divided into 4 groups:≤100 IU/mL,101-1 000 IU/mL,1 001-20 000 IU/mL,and ＞ 20 000 IU/mL,quantitative correlation between two methods was analyzed.Results The linear regression equation of two methods was Y=2.323X-896.3,correlation coefficent r=0.943,P＜0.001.CMIA was as a reference,4 groups were divided for analysis,results showed that when detected specimens was at low concentration of HBsAg,TRIFA value was low compared with CMIA method,while detected specimens was at high concentration of HB sAg,CMIA value was high,two reagents had good consistency in the detection of different concentrations of HBsAg(both P＜0.05),when concentration was at 1 001-20 000 IU/mL,consistency was the best.Conclusion The accuracy of two reagents in the quantitative detection of HBsAg is similar,and the best correlation of detection value is 1 000-20 000 IU/mL.TRIFA assay has wide application for its low-cost and easy to be operated,which is especially suitable for primary hospitals.
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Objective@#To investigate the expression of microRNA-30a (miR-30a) in hepatocellular carcinoma (HCC) and related molecular mechanisms in regulating HCC cell proliferation.@*Methods@#A total of 30 pairs of HCC and adjacent tissue samples were collected, and quantitative real-time PCR and Western blot were used to measure the mRNA and protein expression of forkhead-box protein A1 (FOXA1). Methyl thiazolyl tetrazolium (MTT) assay was used to evaluate the proliferation of HCC cells, luciferase reporter gene assay was performed to verify the target relationship between miR-30a and FOXA1, and MTT assay and Western blot were used to measure the proliferation of HepG2 cells and the protein expression of FOXA1 after miR-30a transfection. The t-test was used for comparison of data between two groups, and a one-way analysis of variance was used for comparison of data between multiple groups. P < 0.05 was considered statistically significant.@*Results@#HCC tissue had significantly lower relative expression of miR-30a than adjacent tissue (1.049 ± 0.380 vs 1.982 ± 1.013, t = 3.985, P < 0.001). At 72 hours after miR-30a overexpression, there was a significant difference in the proliferative capacity of HepG2 cells between the blank control group, the miR-30a-NC group, and the miR-30a group (0.821 ± 0.006 vs 0.816 ± 0.013 vs 0.546 ± 0.020, F = 3.396, P < 0.05), suggesting that miR-30a overexpression significantly inhibited the proliferation of HepG2 cells. FOXA1 was a target gene of miR-30a and its protein expression was negatively regulated by miR-30a, and there was a significant difference in luciferase activity between wild-type and mutant FOXA1-3’UTR vectors (1.221 ± 0.024 vs 2.658 ± 0.031, F = 6.737, P < 0.05). In HepG2 cells, miR-30a overexpression significantly inhibited the protein expression of FOXA1, and there was a significant difference in the relative expression of FOXA1 between the blank control group, the miR-30a-NC group, and the miR-30a group (1.019 ± 0.016 vs 1.022 ± 0.017 vs 0.227 ± 0.021, F = 45.43, P < 0.05). Upregulating the protein expression of FOXA1 reversed the inhibitory effect of miR-30a on the proliferation of HepG2 cells, and there was a significant difference in the proliferative capacity of HepG2 cells between the miR-30a group and the miR-30a+FOXA1 group (0.524 ± 0.023 vs 0.843 ± 0.019, t = 2.507, P < 0.05).@*Conclusion@#MiR-30a exerts its inhibitory effect on the proliferation of HCC cells by negatively regulating the expression of FOXA1.
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OBJECTIVE@#To determine whether time-resolved immunofluorescence assay (TRIFA) shares the similar accuracy and specificity with automatic chemiluminescence immunoassay (CMIA) in analyzing HBeAg levels in hepatitis B. @*METHODS@#A total of 157 serum samples were collected from outpatients with infection of HBV in Xiangya Hospital, Central South University. CMIA and TRIFA were used to analyze HBeAg quantitation and HBeAg/HBeAb qualitative detection, respectively. @*RESULTS@#The linear regression equation for the two methods was Y=0.72779X-4.0551 (r=0.712, P<0.001). Compared with the CMIA, the sensitivity and specificity in detection of HBeAg by TRIFA were 89.89% and 100%, respectively, and the coincidence rate of HBeAg was 94.27% by two assays. Similarly, the sensitivity and specificity in detection of HBeAb by TRIFA were 100% and 95.45%, respectively. The coincidence rate was 97.45% by two assays. @*CONCLUSION@#TRIFA has similar accuracy, sensitivity, and specificity with CMIA in quantitative detection of HBeAg, and their coincidence rate in detection of HBeAg/HBeAb is high.
Subject(s)
Humans , Feasibility Studies , Fluorescent Antibody Technique , Hepatitis B , Hepatitis B Antibodies , Hepatitis B e Antigens , Hepatitis B virusABSTRACT
OBJECTIVE@#To investigate the expression of miR-33b in hepatocellular carcinoma (HCC) and to explore regulatory mechanism of miR-33b for cell proliferation of HCC. @*METHODS@#HCC tissues and adjacent non-tumor tissues were collected for this study (n=32 for each). Real-time PCR and Western blot were conducted to examine the mRNA and protein expression, respectively. MTT assay was used to detect the cell proliferation. Luciferase reporter gene assay was performed to verify the target relationship between miR-33b and Sal-like 4 (SALL4). @*RESULTS@#MiR-33b was significantly downregulated in HCC tissues compared with adjacent non-tumor tissues. Overexpression of miR-33b decreased the proliferation of HCC LH86 cells. SALL4 was identified as a target gene of miR-33b, and its protein expression was negatively regulated by miR-33b. Overexpression of SALL4 reversed the suppressive effect of miR-33b on LH86 cell proliferation. SALL4 was significantly upregulated in HCC tissues compared with adjacent non-tumor tissues. @*CONCLUSION@#The miR-33b suppresses HCC cell proliferation through down-regulation of SALL4.
Subject(s)
Humans , Carcinoma, Hepatocellular , Chemistry , Genetics , Cell Proliferation , Genetics , Physiology , Down-Regulation , Gene Expression Regulation, Neoplastic , Genetics , Physiology , Liver Neoplasms , MicroRNAs , Genetics , Physiology , RNA, Messenger , Real-Time Polymerase Chain Reaction , Transcription Factors , Genetics , Physiology , Tumor Cells, Cultured , Up-RegulationABSTRACT
Objective To evaluate the clinical application of a novel hepatitis B virus YMDD mutation DNA diagnostic kit (magnetic beads method kit).Methods A total of 324 HBV clinical serum samples was tested with the magnetic beads method kit and another kind of fluorescence diagnostic kit (boiling method).Accuracy, specificity, and sensitivity were compared.Results The consistency of positive detection rate of two kits was 100％ (95％ CI : 98.0％ ～ 100％), negative consistency was 97.12％ (95％ CI : 92.8％ ～99.2％) and the total consistency was 98.76％ (95％ CI : 96.9％ ～99.7％).Four cases of discrepant samples were confirmed by sequencing, and statistical analysis performed by Kappa test (Kappa =0.975) shows good consistency between the two methods.Conclusions The magnetic beads method kit has good consistency compared to the regular boiling method kit, and the polymerase chain reaction (PCR) detection system contains an internal positive control (internal control) to avoid a false negative resuit, which is more suitable for clinical diagnosis.
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Objective To investigate the clinical effect of acellular allograft dermal tissue patch used in transvaginal rectovaginal fistula repair.Methods From Jan.2008 to Dec.2011,22 patients with rectovaginal fistula undergoing treatment in Chinese People's Liberation Army General Hospital were studied retrospectively.Twelve patients treated by tissue patch were classified into study group matched with 10 patients with general surgery as controls.Results In study group,11 patients were successfully repaired by their first surgery; one patient was successfully fixed by the second surgery.The successful rate of first operation was 11/12 in study group and 4/5 in recurrent transvaginal rectovaginal fistula.In control group,7 patients were fixed successfully in their first surgeries,the successful rate of first surgery was 7/10.Two primary patients and 1 recurrent patient were successfully fixed by their second surgeries.All of the patients were followed up for (9.0 ± 2.0) months,and no recurrence diseases were observed.Conclusion The transvaginal rectovaginal fistula fixed using acellular allografi dermal tissue patch could get less trauma and higher cure rate.
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<p><b>BACKGROUND</b>To evaluate the response, adverse effects and survival of MVP regimen and TVP regimen.</p><p><b>METHODS</b>Sixty six patients with advanced non-small cell lung cancer were randomized into two groups:MVP arm (32 patients, mitomycin C 6-8 mg/m² d1, vindesine 2-3 mg/m² d1 and d8, cisplatin 70-80 mg/m² d1) and TVP arm (34 patients, pirarubicin 40-50 mg/m² d1, vindesine and cisplatin were the same as arm MVP). Characteristics of the patients were similar in two arms. All patients received two to four cycles of chemotherapy.</p><p><b>RESULTS</b>The overall responses were 34% (11/32) in the MVP arm and 56% (19/34) in the TVP arm. There were 1 complete response, 10 partial responses in the MVP arm and 1 complete response, 18 partial responses in the TVP arm. TVP regimen appeared to have a higher objective response, but no statistically significant difference in the response was observed between two regimens (Chi-square=2.269, P=0.132). Main side effects were hematological toxicities. Grade III+IV hematological toxicities were significantly higher in the patients of arm TVP than arm MVP, especially neutropenia (79% vs 44%, Chi-square=7.458, P=0.006). Median survival time was 12 months vs 8 months, and 1-, 2-, 3-year survival rates were 53% vs 24% (Chi-square=4.943, P=0.026), 17% vs 6%, 6% vs 0, for arm TVP and arm MVP, respectively..</p><p><b>CONCLUSIONS</b>MVP regimen has a lower response rate and longer survival time but less hematological toxicities than TVP regimen. The results suggest MVP regimen is a safe and active regimen for advanced non-small cell lung cancer.</p>