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Objective To investigate the effects of interferon alpha?2b(IFNα?2b)on serum Hepcidin in hepatitis C patients and its mechanism. Methods Hepatitis C patients were divided evenly into treatment group and control group according to whether they had received treatment with IFNα?2b in the past 3 months. The serum hepci?din was compared between the two groups. HepG2 cells and LO2 cells were treated for 24 hours at varied levels of IFNα?2b(0,50,100,200,400μL)and real?time PCR was used to detect the hepcidin,interleukin?6(IL?6)and signal transduction and transcription activator 3(STAT3)mRNA expression of cells. The protein levels of STAT3 and phosphorylated STAT3(pSTAT3)were measured by Western blot. The changes of these indexes were observed with the gradual increase of IFNα?2b levels. Results Serum Hepcidin level in the treatment group was significantly lower than the control(P<0.05). IFNα?2b inhibited the Hepcidin mRNA in HepG2 cells and LO2 cells. pSTAT3 was significantly decreased with the increased levels of IFNα?2b(P<0.05),and the expression of IL?6 and STAT3 had no significant changes with the increase of IFNα?2b. Conclusion The serum Hepcidin levels can be decreased because IFNα?2b suppresses the expression of Hepcidin,and its mechanism may be related with inhibited STAT3 pathway activation.
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Objective To investigate the diagnostic value of laboratory evaluation of renal injury in early diagnosis index. Methods Eighty-six patients in in-patient and out-patient of the department of urology were tested in serum C (Cys C), serum creatinine (Cr) and serum UREA nitrogen (UREA). According to the diagnostics (sixth edition), we evaluated their values for diagnosis of renal injury by analyzing the characteristics of the work curve (ROC). Results According to the renal function indexes, 3 groups were divided such as group A (normal renal function control Ccr 80 mL/min) or the Cys C, Cr, and UREA in (0.47 ± 0.24) mg/L, (85 ± 14) μmol/L, and (4.55 ± 1.33) mmol/L, group B (a decrease in renal function reserves, and renal insufficiency 20 mL/min or less Ccr Cr > UREA. Their correspounding ROC curve (AUC) were in the area of 0.908, and 0.785 and 0817. Cys C had the highest AUC (0.908), and achieved a good diagnosis effect. Conclusion Renal damage laboratory indexes of in the early diagnosis, Cys C has a higher sensitivity and specificity to be worth of clinical promotion.
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Objective To investigate the influence of miR-106b on Wnt/β-catenin pathway in HCC cells. Methods QGY-7703 and HepG2 cells were transfected with miRNA mimics or inhibitors. TOP/FOP luciferase ratio assay was used to test the Wnt/β-catenin pathway activity. The expression of downstream targeted genes of Wnt/β-catenin pathway were examined by Real-time PCR. The accumulation of β-catenin in nuclears were measured by Western blotting. Results Ectopic expression of miR-106b dramatically increased the average TOP/FOP ratio and the mRNA expression of downstream targeted genes in QGY-7703 and HepG2 cells. Compared with that in control cells , miR-106b over-expression promoted the nuclear β-catenin accumulation in QGY-7703 cells. Clonclusion MiR-106b activated Wnt/β-catenin pathway in HCC cells.
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<p><b>OBJECTIVE</b>To investigate the effects of miR-106b expression on the proliferation of human hepatocellular carcinoma cells.</p><p><b>METHODS</b>Real-time PCR was used to detect the expression of miR-106b in human hepatocellular carcinoma (HCC) cell lines and normal liver epithelial THLE3 cells. Over-expression of miR-106b was transfected by miR-106b mimics, and inhibition of miR-106b expression was transfected by miR-106b inhibitors. The effects of miR-106b expression on the proliferation of HCC cells were detected by MTT, clone formation assay and anchorage-independent growth ability assay. Bromodeoxyuridine labeling and flow cytometry analysis were used to examine the effects of miR-106b expression on the cell cycle distribution of the HCC cells.</p><p><b>RESULTS</b>Compared with that in the normal liver epithelial THLE3 cells, the relative expression of miR-106b in HepG2, QGY-7703, BEL-7402, MHCC97H, HCCC-9810, Hep3B, MHCC97L and Huh7 cell lines were 5.37 ± 0.35, 8.45 ± 0.75, 19.22 ± 1.74, 11.93 ± 1.26, 17.03 ± 0.97, 4.19 ± 0.67, 7.94 ± 1.35 and 3.82 ± 0.87, respectively (P < 0.05 for all). Three days after transfection, the miR-106b over-expression was accelerated, while miR-106b inhibitor suppressed the proliferation of HCC cells. The numbers of clones formed were (4.13 ± 0.75) and (3.78 ± 0.47) times higher than that of control cells, and (147.73 ± 15.56) and (138.87 ± 15.58) clones in diameter >1.0 mm were formed by miR-106b-overexpressing cells. When the miR-106b expression was inhibited in the HepG2 and QGY-7703 cells, the clone numbers were (0.18 ± 0.05) and (0.24 ± 0.07) times of that in the controls, and the numbers of clones formed were (23.29 ± 7.14) and (20.60 ± 8.07) (P < 0.05 for all). The positive rates of BrdU labeled miR-106b-overexpressing HepG2 and QGY-7703 cells were (51.89 ± 4.91) % and (54.74 ± 6.10) %, those of the miR-106b-inhibited cells were (6.48 ± 3.15) % and (7.52 ± 2.03) %, significantly different from that in the control cells (P < 0.05 for all). The S phases were dramatically increased from 29.93% and 31.04% to 56.76% and 57.22% in the miR-106b-overexpressing HepG2 and QGY-7703 cells, while they were 19.43% and 19.92% in the miR-106b-inhibited HepG2 and QGY-7703 cells.</p><p><b>CONCLUSIONS</b>MiR-106b overexpression may promote the proliferation and metastasis of HCC cells. The mechanism of this effect may be related to promoting cells into S phase and inhibiting cell apoptosis.</p>
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Humans , Apoptosis , Carcinoma, Hepatocellular , Cell Line, Tumor , Cell Proliferation , Gene Expression Regulation, Neoplastic , MicroRNAs , Metabolism , Real-Time Polymerase Chain Reaction , TransfectionABSTRACT
Objective To explore the value of combined use of laboratory indicators for diagnosis of early renal functional damage. Methods Eighty-six patients with various kidney diseases were enrolled in the Second Affiliated Hospital of Guangzhou Medical University. On admission,the serum Cystatin C(Cys C),creatinine(Cr), Urea,etc were determined. The value of using combined laboratory indicators in the diagnosis of renal functional damage was obtained through the analysis of the receiver operating characteristic curve(ROC curve);multiple variable indicators were grouped to establish multiple logistic regression models to be compared and evaluated. Results In the early and late renal injury groups(group B of 32 cases and group C of 12 cases),the serum levels of Cys C,Cr, and Urea were significantly higher than those in the normal renal function control group(group A of 42 cases),the elevation in level in group C being the most significant〔Cys C(mg/L):3.47±0.75 vs. 1.59±1.29,Cr(μmol/L):669±466 vs. 214±173,Urea(mmol/L):21.22±13.10 vs. 11.04±8.24,PCys C+Urea>Cys C>Cr+Urea(0.920=0.920>0.911>0.908>0.809). In this sequence,the AUC made by Cys C+Cr+Urea and Cys C+Cr were equal,both 0.920, whose sensitivity was 75.0%,specificity 100.0%,positive predictive value 100.0%,negative predictive value 80.0%and diagnostic accuracy rate 87.5%. So,Cys C + Cr combination could be used to substitute Cys C + Cr + Urea, and the former clinical diagnostic effect was the best,much higher than that by using AUC whose curve was made by Cys C alone. Conclusion The value of using only one laboratory indicator for diagnosis of patients with early renal functional damage is not high,while applying Cys C+Cr combination can improve the diagnostic effect greatly,and its sensitivity and specificity are higher.
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Morphological examination of blood cells is an important part of the hematology examination course. In order to enrich teaching resources, network of bilingual education resource was established and put into application by the Department of Hematology in Guangzhou Medical College. The repository improved teaching quality of cell morphology, and played a role in training personnel of hematology examination with solid basic skills.
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Objective To detect the distribution of H.pylori ureA, vacA s1 gene and cagA subtype(ABC, ABD, ABAB, AAD, et al) in patients with digestive diseases in Guangzhou and investigate the relationship with the pathological findings of gastric mucosa.Methods A total of 227 randomly selected gastric mucosa from patients with digestive diseases were enrolled in the research, including 46 without pathological changes, 130 with chronic gastritis, 29 with peptic ulcer, 15 with atrophic gastritis and 7 with gastric cancer.Real-time PCR assay were used to detect Helicobacter pylori ureA gene and vacA s1 gene.EPIYA motifs in the 3′ region of cagA were amplified by conventional PCR followed by subtype sequencing. The conserved gene ureA was used to detect H.pylori infection.Results Among the 227 patients with digestive diseases, 50.7% (115/227) patients were H.pylori positive, in which 91.3%(105/115) carried vacA s1 and 78.3% (90/115) carried cagA. Four types of cagA-EPIYA subtype were detected, including ABC 17.8%(16/90), ABD 78.9%(71/90), AAD 2.2%(2/90) and ABAB 1.1%(1/90).In the non-pathological change group, 32.6% (15/46) were H.pylori positive, in which 28.3% (13/46) carried vacA s1 and 26.1% (12/46) carried cagA;in chronic gastritis group, it was 48.5% (63/130), 43.8% (57/130) and 36.2% (47/130), respectively;in ulcer group, it was 72.4% (21/29), 65.5% (19/29) and 55.2% (16/29), respectively;in atrophic gastritis group, it was 66.7% (10/15), 66.7% (10/15) and 66.7% (10/15), respectively;in gastric cancer group, it was 85.7% (6/7), 85.7% (6/7) and 71.4% (5/7), respectively.The distribution of H.pylori among the 4 groups had statistical significance (χ2=16.72;P<0.01). H.pylori was more prevalent in ulcer, atrophic gastritis and cancer group than in inflammation group and non-pathological change group (χ2=16.02;P<0.01).In patients infected by H.pylori, there was no significant difference in the distribution of vacA s1 gene as high virulence factors among non-pathological change, inflammation, ulcer, atrophic gastritis and cancer group (χ2=2.00;P=0.74), as well as cagA (χ2=3.44;P=0.49) and EPIYA subtypes (χ2=3.66;P=0.45).Conclusions H.pylori infection is significantly associated with the pathological change of gastric mucosa for patients with digestive diseases in Guangzhou, while the relationship with the pathogenicity of H.pylori with high virulence genotype is not significant.More samples and diseases reclassification are needed to make an advanced analysis of the effect of H.pylori with high virulence in gastrointestinal diseases development.