RESUMEN
ObjectiveTo investigate the effect of oxaliplatin on the activation of hepatic stellate cells （HSCs）， as well as the association of oxaliplatin with microRNA-30a-5p and autophagy. MethodsHSC-LX2 cells were cultured and divided into groups according to the following three protocols： control group， PDGF treatment group， oxaliplatin treatment group， oxaliplatin+PDGF treatment group； control group， microRNA-30a-5p transfection group， PDGF treatment group， microRNA-30a-5p transfection+PDGF treatment group； control group， 3-MA group， microRNA-30a-5p inhibitor group， microRNA-30a-5p inhibitor+3-MA group. Western Blot was used to measure the expression of HSC activation-related proteins （Collagen-I and alpha-smooth muscle actin ［α- SMA］） and HSC autophagy-related proteins （Beclin-1， P62， and LC3B）； LysoTracker staining and immunofluorescence assay were used to measure the expression of LC3B autophagosomes； RT-PCR was used to measure the expression level of microRNA-30a-5p； bioinformatics techniques were used to predict the potential targets of microRNA-30a-5p in HSCs. The independent-samples t test was used for comparison of normally distributed continuous data between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsAfter the cells were treated with oxaliplatin， RT-PCR results showed that the oxaliplatin treatment group had a significantly higher expression level of microRNA-30a-5p than the control group （P<0.01）； Western Blot showed that the oxaliplatin treatment group had significant reductions in the expression levels of the HSC activation-related proteins α-SMA and Collagen-Ⅰ and the autophagy-related proteins Beclin 1 and LC3BⅡ/Ⅰ （all P<0.001）； immunofluorescence assay showed that the oxaliplatin treatment group had a significantly lower number of autophagosomes than the control group （P<0.05）. After HSC-LX2 cells were transfected with microRNA-30a-5p mimic， compared with the control group， the microRNA-30a-5p mimic group had significant reductions in the expression levels of the autophagy-related proteins Beclin 1 and LC3BⅡ/Ⅰ （P<0.05） and the HSC activation-related protein Collagen-Ⅰ （P<0.001）； after HSC-LX2 cells were transfected with microRNA-30a-5p inhibitor， Western Blot showed that compared with the control group， the microRNA-30a-5p inhibitor group had significant increases in the expression levels of the HSC activation-related proteins Collagen-Ⅰ and α-SMA and the autophagy-related protein Beclin 1 （t=2.41， 2.32， and 4.57， all P<0.05）. Western Blot showed that compared with the control group， the microRNA-30a-5p inhibitor group had significant increases in the expression levels of the HSC autophagy-related protein Beclin 1 and the HSC activation-related protein α-SMA （both P<0.05）， and after the treatment with the autophagy inhibitor 3-MA， there were no significant differences in the expression of these proteins between the two groups （P>0.05）. The bioinformatics analysis using TargetScan， PicTar， and miRanda databases showed that the autophagy-related protein Beclin-1 might be a potential target of miRNA-30a-5p. ConclusionOxaliplatin can inhibit the activation of HSCs by upregulating the expression of microRNA-30a-5p， which provides new ideas and a new target for the treatment of liver fibrosis.
RESUMEN
Objective:To analyze and screen microRNA (miRNA) related to the prognosis of gastric cancer(GC) by bioinformatics analysis, and to construct and validate a risk score model.Methods:The human genome miRNA sequencing data and corresponding clinicopathological data of the 491 samples (446 GC tissue samples and 45 normal gastric tissue samples) were downloaded from the cancer genome atlas (TCGA) database. The differentially expressed microRNA (DEM) was analyzed with edgeR package of R 4.0.2 software and the obtained DEM’s profile was randomly divided into training set and test set according to the ratio of 1∶1. The miRNA related to prognosis were analyzed and screened with univariate Cox regression and least absolute shrinkage and selection operator (LASSO) regression, and multivariate Cox regression analysis was further performed to analyze the screened prognostic-related miRNA and then the prognostic risk score model was constructed. Kaplan-Meier curve, receiver operating characteristic curve (ROC), and dynamic area under the ROC were drawn to evaluate the predictive power of the model.Results:A total of 175 DEM in GC tissues were screened out based on the cut-off criteria of |log2 Fold Change|>1.5 and P<0.01. Six DEMs related to the overall survival rate of patients with GC were screened out by univariate Cox regression and LASSO regression analysis, and then a five-miRNA risk score model was successfully constructed by multivariate Cox regression. The risk score=0.183×hsa-miRNA-184+ 0.086×hsa-miRNA-675-0.231×hsa-miRNA-2115+ 0.548×hsa-miRNA-3943-1.455×hsa-miRNA-1246. In the training set, test set and overall data set, the cumulative survival rates of the patients with higher risk score were lower than those of the patients with lower risk score, respectively, and the differences were statistically significant ( χ2=18.90, 9.50 and 26.70, all P<0.05). The prediction power of the model was better than that of TNM stage. And the results of stratified analysis showed the predictive ability of the model in patients with early GC. The results of univariate Cox regression and multivariate Cox regression demonstrated that the risk score of the model, gae and M stage were independent risk factors for poor prognosis in patients with GC (hazard ratio(95% confidence interval)1.19(1.07 to 1.32), 1.20(1.06 to 1.40), 1.50(1.01 to 2.23), 1.90(1.28 to 2.90), 1.34(1.15 to 1.57), 2.10(1.05 to 4.40); all P<0.05). Conclusion:The 5-miRNA risk score model based on 5 miRNAs which was an independent prognostic factor had high accuracy in predicting the prognosis of patients with GC.
RESUMEN
The pathological basis of liver fibrosis is the deposition of extracellular matrix (ECM), and myofibroblasts are the main source of ECM. Epithelial-mesenchymal transition (EMT) is one of production mechanisms of myofibroblasts. At present, a large number of studies have shown that intervention of key EMT molecules and signaling pathways as targets can reduce liver fibrosis. Based on literature review, this article summarizes the signaling pathways associated with EMT, important regulatory molecules, and drugs targeting EMT in the treatment of liver fibrosis, so as to provide new ideas for the treatment of liver fibrosis.
RESUMEN
Chronic liver diseases have various etiologies and often have poor long-term prognosis in clinical practice. Y-box binding protein-1 (YB-1) is a multifunctional protein, and in-depth studies in recent studies have found that it plays a key role in the development and progression of chronic liver diseases such as liver fibrosis and hepatocellular carcinoma (HCC). This article summarizes the role of YB-1 in chronic liver diseases such as liver fibrosis, HCC (proliferation, apoptosis, metastasis, prognosis, and drug resistance), and liver failure, so as to provide a theoretical basis for the diagnosis and treatment of chronic liver diseases.
RESUMEN
ObjectiveTo screen out the microRNAs (miRNAs) associated with the prognosis of hepatocellular carcinoma (HCC) through data mining of miRNA transcriptome data of HCC downloaded from The Cancer Genome Atlas (TCGA) database, to establish a miRNA risk score model, and to investigate its value in predicting the prognosis of HCC. MethodsThe miRNA expression data and clinical data of HCC samples were downloaded from TCGA database and R language was used to screen out differentially expressed miRNAs between HCC tissue and adjacent tissue, which were randomly divided into training set and testing set after being integrated into clinical data. Univariate Cox regression analysis and least absolute shrinkage and selection operator (LASSO) Cox regression analysis were performed for the training set to screen out the miRNAs associated with the prognosis of HCC, and then a miRNA risk score model was established. The Kaplan-Meier method was used to evaluate the robustness of the model and whether it could predict the prognosis of patients in the same clinical stage. Finally, the receiver operating characteristic (ROC) curve was plotted and the area under the ROC curve (AUC) was calculated to compare the predictive accuracy of the model versus TNM staging in the training set, the testing set, and the entire set. ResultsA total of 300 differentially expressed miRNAs were screened out and the LASSO Cox regression analysis revealed that hsa-miR-139-5p, hsa-miR-1180-3p, hsa-miR-1269b, hsa-miR-3680-3p, hsa-miR-509-3-5p, and hsa-miR-31-5p were associated with the prognosis of HCC. The risk score was calculated for each sample according to the established miRNA risk score model, and the samples were divided into high-risk group and low-risk group according to the median risk score. The Kaplan-Meier curve showed that in both training and testing sets, the high-risk group had a significantly lower survival rate than the low-risk group (P＜0.05). The ROC curve was used to evaluate the prediction efficiency of this model, and the results showed that in the training set, the testing set, and the entire set, the miRNA model had an AUC of 0.817, 0.808, and 0.814, respectively, while TNM staging had an AUC of 0.667, 0.665, and 0.663, respectively. The results of independent prognostic analysis also showed that this miRNA score model could be used as an independent prognostic factor for HCC (P＜0.05). ConclusionHsa-miR-139-5p, hsa-miR-1180-3p, hsa-miR-1269b, hsa-miR-3680-3p, hsa-miR-509-3-5p, and hsa-miR-31-5p are associated with the prognosis of HCC, and the miRNA risk score model has a better prediction accuracy than TNM staging in the training set, the testing set, and the entire set. The stratified analysis also shows that the model can predict the prognosis of patients within the same TNM stage, and therefore, it has a certain reference value in clinical practice and can be used as an independent model for predicting the prognosis of HCC patients.
RESUMEN
Objective To study the changes of plasma endotoxin and procalcitonin in patients with esophagogastric varices and provide a theoretical basis for prophylactic antibiotics after endoscopic treatment. Methods Fifty cases of patients with esophageal and gastric varices accepted the endoscopic treatment.The patients were divided into antibiotic group (32 cases)and non-antibiotic group (18 cases).The plasma endotoxin and procalcitonin were measured before and on the first day and 7th day after endoscopic treatment.Results The plasma levels of endotoxin and procalcitonin were not significantly different on the first and 7th day after endoscopic treatment compared with preoperative levels in antibiotic group.But in non-antibiotic group,the levels significantly increased on 7th day after endoscopic treatment compared with preoperative levels (P <0.05).And in patients of Child-Pugh A grade,the level of plasma procalcitonin significantly increased on 7th day after endoscopic treatment compared with preoperative levels (P <0.01), but the procalcitonin was not significantly different on the first and 7th day after operation.And in patients of Child-Pugh B and C grades,the levels of plasma endotoxin and procalcitonin significantly increased on the 7th day(P <0.01).Conclusion The levels of plasma endotoxin and procalcitonin in non-antibiotic group increase after endoscopic treatment,which suggests the risk of infection.Prophylactic antibiotics after endo-scopic treatment should be considered for the patients of Child-Pugh B and C grades.
RESUMEN
Objective To explore the expression of heme oxygenase-1 (HO-1) in gastric mucosa of patient with portal hypertensive gastropathy (PHG),and its role in the pathogenesis of PHG.Methods The specimens were obtained from the gastric mucosa of 22 healthy subjects,20 portal hypertension (PHT) without PHG,and 22 PHG patients.Histological changes of the gastric mucosa were detected,and portal venous flow (PVF) was measured.The expression of HO-1 protein in gastric mucosal specimens was assessed by immunohistochemistry and Western blot analysis,respectively.The relationship between HO-1 expression and PHG severity score,HO-1 expression and PVF,PHG severity score and clinical parameters were investigated.Results HO-1 protein expression in gastric mucosa of PHG and PHT was significantly higher than that of the control (P ＜ 0.05),while there was no significant difference in this variable between PHG and PHT (P ＞ 0.05).The positive correlation was detected between HO-1 expression and PHG severity score in PHG patients (r =0.459,P ＜0.05),however,PHG severity score was irrelevant to severity of esophageal varice (r =0.059,P ＞ 0.05) or to Child-Pugh classification (r =-0.001,P ＞ 0.05).Of all the patients with PHT and PHG,no significant correlation was found between HO-1 expression and PVF (r =0.071,P ＞ 0.05).Conclusion HO-1 protein is up-regulated in gastric mucosa of PHG patients,which may contribute to gastric circulation disorder of PHG.
RESUMEN
Objective To investigate hepatic expressions and significances of cannabinoid receptor 1 (CB1) and cannabinoid receptor 2(CB2) in C57 mice with experimental cirrhosis.Methods Thirty C57 mice were randomly divided into three groups,i.e.normal control group,model control group and model colchicine group.Hepatic fibrosis model was prepared by intraperitoneal injection of carbon tetrachloride.The expressions of CB1 and CB2 in liver tissue of mice were observed by immunohistochemistry.The scores of inflammation grade (G) and fibrosis stage (S) were simultaneously performed.Results The scores of G and S in model control group and model colchicine group were significantly higher than those in normal control group( F =125.41,P =0.00; F =99.18,P =0.00).The scores of G and S in model control group were significantly higher than those in model colchicine group(P ＜0.01 ).The scores of CB1 and CB2 expressions in model control group and model colchicine group were significantly higher than those in normal control group ( F =29.27,P =0.00; F =36.99,P =0.00).The scores of CB1 and CB2 in model control group were significantly higher than those in model colchicine group( P ＜ 0.05 or P ＜ 0.01 ).There were significant relationships among scores of CB1,CB2,G and S in model control group and model colchicine group(Ps ＜0.05).As the scores of G and S became higher,the expressions of CB1 and CB2 gradually became more intensive.Conclusion The hepatic expressions of CB1 and CB2 in C57 mice with experimental cirrhosis increased significantly and have significant relationship with the grades of liver tissue inflammation and fibrosis.
RESUMEN
Objective To explore the dynamic expression of ROCK-Ⅰ,p-MBS Thr-697,α-SMA protein and their mRNA in the hepatic fibrogenesis and the changes of actin cytoskeleton.Methods ROCK-Ⅰ and p-MBS Thr-697 protein in liver were determined by Western blot and their mRNA was examined by reverse transcription-polymerase chain reaction (RT-PCR),while the distribution of ROCK-Ⅰ and α-SMA in liver was assessed immunohistochemis-tically.The change of actin cytoskeleton was shown by fluorescence.Results With the development of hepatic fi-brosis,the positive areas in model groups at week 1 to 4 of ROCK-Ⅰ and α-SMA of the rat livers were larger than that in control group respectively(P <0.05).ROCK-Ⅰ,p-MBS Thr-697 ,α-SMA protein and mRNA were increased than that in control group respectively.ROCK-Ⅰ mRNA expression correlated with α-SMA (r =0.718,P <0.05).With the development of liver fibrosis,the images of fluorescence were inhanced.Conclusion With the develop-ment of liver fibrosis,both protein and mRNA of ROCK-Ⅰ increased.
RESUMEN
Objective The primary aim of this study was to examine the proportion and natural history of Helicobacter pylori (Hp) negative bleeding peptic ulcers. Methods The study was designed as a multiple-center, case-control study conducted in 14 endoscopy centers in China from April 2006 to March 2007. Each center was expected to recruit 30 peptic ulcer patients with bleeding ( PUB group) and 30 without (PU group). All screened patients with upper gastrointestinal bleeding received endoscopy within 24 hours of admission. Biopsy specimens were taken from the antrum to determine Hp infection by rapid urease test and pathology. Patients with negative Hp infection at first examination were asked to receive urease breathe test (UBT) one month later. Results A total of 617 patients were enrolled with 263 in PUB group and 354 in PU group. There is no significant difference in demographic characters between 2 groups ( P ＞0. 05). The rate of Hp infection in PUB group ( 161/263, 61.2% ) was significantly lower than that in PUgroup (311/354, 87. 9%, P ＜0. 001 ). The incidence of complex ulcer in Hp positive PUB patients was 7.5% ( 12/161 ), which is significantly higher than that in Hp negative PUB patients ( 1/102, 1.0% , P =0. 018). In PUB group, no significant differences were found between Hp positive and negative patients in regarding of age, sex, rates of haematemesis, duodenal ulcer and gastric ulcer, and size of ulcer ( P ＞0. 05 ). Among 102 Hp negative cases in PUB, no positive case was found in UBT one month later. Conclusion We have demonstrated a rise in the incidence of Hp negative bleeding ulcers in China. The idiopathic ulcer was not rare, and might have a higher tendency to cause bleed.
RESUMEN
Objective To investigate the effects of over-expression of wild-type PTEN gene and its mutant (G129E) on apoptosis and proliferation of activated hepatic stellate cells (HSC) in vitro and its potential mechanisms. Methods The activated HSC cells were cultured in vitro and transfected with PTEN gene and G129E gene via adenoviral vector. The apoptosis of HSC was measured by MTT , and its proliferation was assessed by TUNEL and flow cytometry (FCM) . Western blotting and real-time fluorescent quantitation PCR were used to detect expression of PTEN in HSC. And the changes of Bcl-2 and Bax expression were tested by Western blotting. Results The wild type PTEN gene and G129E gene were successfully transducted into HSC, which resuted in elevated expression of Bax and reduced expression of Bcl-2 (P<0.01). After transduction, HSC proliferation was markedly inhibited with inhibitory rates of 14.03% and 23.12% at 48 and 72 hours in Ad-PTEN ,respectively, as well as 9.52% and 12.63% in Ad-G129E, respectively. Apoptotic rate of HSC exposed to Ad-PTEN or Ad-G129E for 72 hours increased significantly (P<0.01). Furthermore, wild type PTEN was more powerful than G129E for above-mentioned effects. Conclusions Over-expression of wild type PTEN and its mutant G129E can inhibit the proliferation of activated HSC, and induce HSC apoptosis through the Bcl-2/Bax pathway. In addition, the effect of wild type PTEN is more powerful than that of G129E.
RESUMEN
Objective To investigate the expression of TLR2 in colon mucosa of ulcerative colitis (UC) and to analyze the correlation with clinical activity and endoscopic grading. Methods The biopsies from 47 UC patients and 13 healthy controls were collected, and the expression of TLR2 protein and mRNA in colonic mucosa was determined by Western Blot and semi-quantitative RT-PCR, respectively. The patients were graded according to endoscopic and clinical findings. Results The expressions of TLR2 protein and TLR2 mRNA in UC patients were significantly increased than those in healthy controls, which was correlated with the progression of the disease. Conclusion The expressions of TLR2 protein and TLR2 mRNA in colon mucosa from UC patients might be used as a marker for disease activity.
RESUMEN
Objective To evaluate the effect of osteopontin(OPN) on the adhesion and migration of hepatic stellate cells(HSC) and focal adhesion kinase(FAK)phosphorylation.Methods The proliferation of HSCs was determined by MTT.The adhesion rates were shown by toluidine blue colorimetric assay.The HSC migration rates were examined by HE staining.The expression of FAK phosphorylation was examined by Western blotting analysis.Results ①Compared with control group,OPN at concentrations of 8 mg/L,16 mg/L and 32 mg/L promoted the adhesion of HSCs(P
RESUMEN
Objective To observe the expressive changes of RhoA and phosphorylated myosin light chain in rat liver tissue undergoing hepatic fibrogenesis.Methods The liver histopathological changes were evaluated by hematoxylin and eosin staining and Masson's trichrome method.Western blot was used to determine the expressions of RhoA and p-MLC(Thr18/Ser19),and reverse transcription-polymerase chain reaction(RT-PCR) to determine the expression of RhoA mRNA.Results With the development of hepatic fibrogenesis,the protein expressions of RhoA and p-MLC(Thr18/Ser19) and the mRNA expression of RhoA were significantly increased.RhoA and p-MLC(Thr18/Ser19)correlated with ?-SMA positively,respectively.Conclusion Rho/ROCK signaling pathways are changed in the process hepatic fibrogenesis.
RESUMEN
Objective By observing the changes in NGF and its receptor P75 in liver of CCl4-induced toxic rats and to evaluate the role of NGF in the mechanism of hepatic fibrosis.Methods The expressions of NGF mRNA and its receptor P75 were detecteded by both hemi-quantitative RT-PCR and Western blot analysis.Results The expressions of NGF mRNA and its receptor P75 in liver of CCl4-induced toxic rats at 24th hour group were higher than that in control group(P
RESUMEN
Objective To identify the genotype of clinical stenotrophomonas maltophilia(SMA) isolates and investigate the characteristics of SMA in nosocomial infections.Methods Totally 165 strains of SMA were clinically isolated during the period of 2004 to 2007.Disc diffusion test(K-B method) was used for antibiotic susceptibility.qacE△1 gene was detected by polymerase chain reation(PCR).The gene homology in the SMA strains was analyzed by pulsed field gel electrophoresis(PFGE).Results Among the tested SMA strains 87.9% sourced from low respiratory tract infection.The antibiotics with more than 80% of sensitive rate against SMA were minocycline,trimethoprim-sulfamethoxazole and levofloxacin.The positive rate of qacE△1 gene was 13.3% in 60 tested strains.The analysis of gene homology for the 11 clinical strains showed that two genotypes from identical clone were found in both respiratory ICU and emergency ICU respectively.Conclusions SMA was an important pathogenic bacterium in nosocomial infections.The treatment for SMA infection is very difficult since its multi-drug resistance.More attention for effective sterilization and isolation of patients must be paid to prevent the transmission of SMA from same clone.
RESUMEN
OBJECTIVE To find out the clinical characteristics of nosocomial infections with Stenotrophomonas maltophilia(SMA) and investigate the disinfectant resistant gene of qacE△1,improve the diagnosis and decrease of nosocomial infection cases.METHODS Toally 165 strains of SMA were clinically isolated from 2004 to 2007.The gene of qacE△1 was analyzed by polymerase chain reation(PCR) and homology of the strains was analyzed by the method of by pulsed field gel electrophoresis(PFGE) genotype.RESULTS Susceptible factors were old age,seriousness of underlying disease,prolonged hospitalization and invasive operation with infection of SMA.The lower respiratory tract infection was mostly common with SMA,rate of which was 87.9%.Antibiotic sensitive rate more than 80% against SMA was minocycline,trimethoprim-sulfamethoxazole and levofloxacin.The positive rate of gene qacE△1 was 13.3% in 60 strains tested.Two genotypes in SMA strains from respiratory ICU(RICU) were with the same clone.This result proved that clone transmission occurred in RICU.CONCLUSIONS SMA is an important nosocomial infection pathogen.With the multi-drug resistance,the therapy of infection with SMA is very hard in clinic.As result of gene of qacE△1 and the same clone transmission,clinicians should play an important role of surveillance of effect of disinfection.
RESUMEN
Objective: To investigate the management of NH~-_3 burned cases with inhalation injury. Methods:The clinic treatments of 15 NH~-_3 burned cases complicated with inhalation injury in his hospital between January 1998 and April 2004 were analyzed retrospectively.His management included reasonable fluid therapy,prompt and efficient usage of antibiotics ,tracheotomy after diagnosis,mechanical ventilation,bronchoalveolar lavage using fibrobrochoscope after the shock period and short-time usage of glucocorticoid. Results: Of the 15 cases,14 patients were cured and 1 patient was improved without any death.The cure rate was 93.3%. Conclusion: All of the managements should be performed as quickly as possible.Paying much attention to prevent the complications could improve the cure rate.Tracheotomy and bronchoalveolar lavage in the early stage are vital to patients.
RESUMEN
0.05),but positively on PBD 7(r= 0.890,P
RESUMEN
Objective:To investigate the distribution and evolution of immunocytes in infantile hemangioma(IH).Methods:Fifty-two infantile hemangioma samples were investigate.The distribution of CD3+T cells,CD8+T cells and S-100+dendritic cell(DC) in IH was observed with.Results:CD3+T cell was not found among the earliest IH.In the middle proliferating stage,the number of CD3+T cells increased;But the CD8+T cells were still scare.In the late proliferating stage,there were many CD3+T cells and the number of CD8+T cells also increased.In the early involuting stage,there were still a number of CD3+ and CD8+ T cells around the microvessels.In the middle involuting and involuted stage,only a few of CD3+T cells and CD8+ T cells existed.In the early proliferating stage,there were some DC in IH.During the middle and late proliferating stage,the number of DC increased significantly.Since the early involuting stage,the DCs decreased rapidly and disappeared.Conclusion:The distribution of T cells and its subsets and DC have close relationship with the pathologic evolution of infantile hemangioma.