Individual mortality risk predictive system of patients with acute-on-chronic liver failure based on a random survival forest model / 中华医学杂志(英文版)

BACKGROUND@#The basis of individualized treatment should be individualized mortality risk predictive information. The present study aimed to develop an online individual mortality risk predictive tool for acute-on-chronic liver failure (ACLF) patients based on a random survival forest (RSF) algorithm.@*METHODS@#The current study retrospectively enrolled ACLF patients from the Department of Infectious Diseases of The First People's Hospital of Foshan, Shunde Hospital of Southern Medical University, and Jiangmen Central Hospital. Two hundred seventy-six consecutive ACLF patients were included in the present study as a model cohort (n = 276). Then the current study constructed a validation cohort by drawing patients from the model dataset based on the resampling method (n = 276). The RSF algorithm was used to develop an individual prognostic model for ACLF patients. The Brier score was used to evaluate the diagnostic accuracy of prognostic models. The weighted mean rank estimation method was used to compare the differences between the areas under the time-dependent ROC curves (AUROCs) of prognostic models.@*RESULTS@#Multivariate Cox regression identified hepatic encephalopathy (HE), age, serum sodium level, acute kidney injury (AKI), red cell distribution width (RDW), and international normalization index (INR) as independent risk factors for ACLF patients. A simplified RSF model was developed based on these previous risk factors. The AUROCs for predicting 3-, 6-, and 12-month mortality were 0.916, 0.916, and 0.905 for the RSF model and 0.872, 0.866, and 0.848 for the Cox model in the model cohort, respectively. The Brier scores were 0.119, 0.119, and 0.128 for the RSF model and 0.138, 0.146, and 0.156 for the Cox model, respectively. The nonparametric comparison suggested that the RSF model was superior to the Cox model for predicting the prognosis of ACLF patients.@*CONCLUSIONS@#The current study developed a novel online individual mortality risk predictive tool that could predict individual mortality risk predictive curves for individual patients. Additionally, the current online individual mortality risk predictive tool could further provide predicted mortality percentages and 95% confidence intervals at user-defined time points.

Prevalence and clinical significance of metabolic disorders of lipids, glucose and uric acid at different fibrosis stages of patients with chronic hepatitis B / 中华传染病杂志

Objective To explore the prevalence and clinical significance of metabolic disorder of lipids, glucose and uric acid at different fibrosis stages of patients with chronic hepatitis B (CHB).Methods From January 2006 to December 2014, 1 812 CHB patients in Department of Infectious Diseases, Shunde Hospital of Southern Medical University were retrospectively enrolled and analyzed.All biochemistry indexes were obtained by automatic biochemical instrument.Polymerase chain reaction was used for detecting serum hepatitis B virus (HBV) DNA, and particles immune detection kit was used for detecting hepatitis B e antigen (HBeAg).In statistical analyses, chi-square test, nonparametric test and Logistic analysis were used.Results The metabolic disorder prevalence in 1 812 CHB patients was as follows, 455 cases (25.1%) with decreased high density lipoprotein, 435 cases (24.0%) with increased uric acid, 342 cases (18.9%) with increased total cholesterol, 254 cases (14.0%) with increased triglyceride, 171 cases (9.4%) with decreased apolipoprotein A, 165 cases (9.1%) with increased apolipoprotein B, 162 cases (8.9%) with increased low density lipoprotein and 117 cases (6.5%) increased fasting blood glucose.Patients who had mild liver fibrosis tended to have metabolic disorders of uric acid (26.4%), total cholesterol (22.8%) and high density lipoprotein cholesterol (20.5%).Patients who had moderate liver fibrosis tended to have metabolic disorders of high density lipoprotein (27.2%) and uric acid (20.9%).Patients who had severe liver fibrosis tended to have metabolic disorders of high density lipoprotein (33.6%) and uric acid (22.2%).Multivariate Logistic regression analysis showed that inflammation activty (OR=17.31, 95% CI: 13.410-22.336, P=0.001), age (OR=1.019, 95%CI:1.005-1.035, P=0.010), sex (OR=1.497, 95% CI: 1.061-2.111, P=0.022), apolipoprotein A (OR=0.50, 95% CI: 0.281-0.892, P=0.019) and HBV DNA (OR=0.904, 95% CI: 0.858-0.952, P=0.001) may be independent predictors of moderate and severe liver fibrosis.Conclusions CHB patients with mild liver fibrosis tend to have metabolic disorders of uric acid, total cholesterol and high density lipoprotein cholesterol;patients with moderate liver fibrosis tend to have metabolic disorders of high density lipoprotein and uric acid;and patients with severe liver fibrosis tend to have metabolic disorders of high density lipoprotein and uric acid.

Rac1+ cells distributed in accordance with CD 133+ cells in glioblastomas and the elevated invasiveness of CD 133+ glioma cells with higher Rac1 activity / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Recent studies have suggested that cancer stem cells are one of the major causes for tumor recurrence due to their resistance to radiotherapy and chemotherapy. Although the highly invasive nature of glioblastoma (GBM) cells is also implicated in the failure of current therapies, it is not clear how glioma stem cells (GSCs) are involved in invasiveness. Rac1 activity is necessary for inducing reorganization of actin cytoskeleton and cell movement. In this study, we aimed to investigate the distribution characteristics of CD133+ cells and Rac1+ cells in GBM as well as Rac1 activity in CD133+ GBM cells, and analyze the migration and invasion potential of these cells.</p><p><b>METHODS</b>A series of 21 patients with GBM were admitted consecutively and received tumor resection in Tianjin Medical University General Hospital during the first half of the year 2011. Tissue specimens were collected both from the peripheral and the central parts for each tumor under magnetic resonance imaging (MRI) navigation guidance. Immunohistochemical staining was used to detect the CD133+ cells and Rac1+ cells distribution in GBM specimens. Double-labeling immunofluorescence was further used to analyze CD133 and Rac1 co-expression and the relationship between CD133+ cells distribution and Rac1 expression. Serum-free medium culture and magnetic sorting were used to isolate CD133+ cells from U87 cell line. Rac1 activation assay was conducted to assess the activation of Rac1 in CD133+ and CD133 - U87 cells. The migration and invasive ability of CD133+ and CD133 - U87 cells were determined by cell migration and invasion assays in vitro. Student's t-test and one-way analysis of variance (ANOVA) test were used to determine statistical significance in this study.</p><p><b>RESULTS</b>In the central parts of GBMs, CD133+ cells were found to cluster around necrosis and occasionally cluster around the vessels under the microscope by immunohistological staining. In the peripheral parts of the tumors, CD133+ cells were lined up along the basement membrane of the vessels and myelinated nerve fibers. Rac1 expression was high and diffused in the central parts of the GBMs, and the Rac1+ cells were distributed basically in accordance with CD133+ cells both in the central and peripheral parts of GBMs. In double-labeling immunofluorescence, Rac1 was expressed in (83.14 ± 4.23)% of CD133+ cells, and CD133 and Rac1 co-expressed cells were located around the vessels in GBMs. Significantly higher amounts of Rac1-GTP were expressed in the CD133+ cells (0.378 ± 0.007), compared to CD133- cells (0.195 ± 0.004) (t = 27.81; P < 0.05). CD133+ cells had stronger ability to migrate (74.34 ± 2.40 vs. 38.72 ± 2.60, t = 42.71, P < 0.005) and invade (52.00 ± 2.28 vs. 31.26 ± 1.82, t = 30.76, P < 0.005), compared to their counterpart CD133- cells in transwell cell migration/invasion assay.</p><p><b>CONCLUSIONS</b>These data suggest that CD133+ GBM cells highly express Rac1 and have greater potential to migrate and invade through activated Rac1-GTP. The accordance of distribution between Rac1+ cells and CD133+ cells in GBMs implies that Rac1 might be an inhibited target to prevent invasion and migration and to avoid malignant glioma recurrence.</p>

Glioma stem cells enhanced angiogenesis and its relationship with microvessel / 中华外科杂志

<p><b>OBJECTIVES</b>To dynamically observe how glioma stem cells promote the tumor formation and angiogenesis, and to study the correlation between the distribution of glioma stem cells and microvessels within different growth stages of subcutaneous tumor.</p><p><b>METHODS</b>Stem cell medium culture and magnetic activated cell sorting were carried out to obtain CD133+ cells from C6 rat glioma cell line. Sprague Dawley (SD) rat ears model were established to observe glioma stem cells promoting blood vessel formation. Subcutaneous glioma model of C6 and immunohistochemical staining of hypoxia inducible factor-1α (HIF-1α) and CD133 were used to investigate the relationship between distribution of glioma stem cells and microvessels. Expressions of CD133 protein in each stage of the subcutaneous tumor were detected by Western blot.</p><p><b>RESULTS</b>Isolation and identification of glioma stem cells deprived from C6 glioma cell line successfully, the establishment of ears model showed real-time dynamic observation of CD133+ cells involved in angiogenesis and tumor formation. SD rat model of subcutaneous glioma showed the initial of tumor formation, CD133+ cells scattered. With tumor growth, CD133+ cells began to tend to capillaries, in late distributed clusters in perivascular. Meanwhile as tumor growth, CD133 protein expression was gradually increased: the values of Western blot analysis of CD133 expression on 6, 9, 12, 15, 20 d were 0.208±0.004, 0.282±0.003, 0.360±0.004, 0.564±0.135, 0.756±0.007, the differences were significant between different groups (F=2601.681, P<0.01). At a high magnification, the CD133 scores with immunohistochemical staining on 6, 9, 12, 15 d were 0.8±0.4, 2.4±0.5, 4.0 ± 0.7, 6.0±0.7; HIF-1α scores were 0.8±0.4, 2.8±0.8, 5.0±0.7, 6.8±0.4. By Spearman rank correlation analysis found that the relationship between CD133 and HIF-1α expression was positively correlated (r=0.921, P<0.01).</p><p><b>CONCLUSIONS</b>Glioma stem cells promote angiogenesis more than non-stem cells; HIF-1α and its downstream gene product might mediate the distribution of glioma stem cells around the perivascular.</p>

Enhanced invasion in vitro and the distribution patterns in vivo of CD133+ glioma stem cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Recent studies have suggested that cancer stem cells cause tumor recurrence based on their resistance to radiotherapy and chemotherapy. Although the highly invasive nature of glioblastoma cells is also implicated in the failure of current therapies, it is not clear whether cancer stem cells are involved in invasiveness. This study aimed to assess invasive ability of glioma stem cells (GSCs) derived from C6 glioma cell line and the distribution patterns of GSCs in Sprague-Dawley (SD) rat brain tumor.</p><p><b>METHODS</b>Serum-free medium culture and magnetic isolation were used to gain purely CD133(+) GSCs. The invasive ability of CD133(+) and CD133(-) C6 cells were determined using matrigel invasion assay. Immunohistochemical staining for stem cell markers and luxol fast blue staining for white matter tracts were performed to show the distribution patterns of GSCs in brain tumor of rats and the relationship among GSCs, vessels, and white matter tracts. The results of matrigel invasion assay were estimated using the Student's t test and the analysis of Western blotting was performed using the one-way analysis of variance (ANOVA) test.</p><p><b>RESULTS</b>CD133(+) GSCs (number: 85.3 ± 4.0) were significantly more invasive in vitro than matched CD133(-) cells (number: 25.9 ± 3.1) (t = 14.5, P < 0.005). GSCs invaded into the brain diffusely and located in perivascular niche of tumor-brain interface or resided within perivascular niche next to white fiber tracts. The polarity of glioma cells containing GSCs was parallel to the white matter tracts.</p><p><b>CONCLUSIONS</b>Our data suggest that CD133(+) GSCs exhibit more aggressive invasion in vitro and GSCs in vivo probably disseminate along the long axis of blood vessels and transit through the white matter tracts. The therapies targeting GSCs invasion combined with traditional glioblastoma multiforme therapeutic paradigms might be a new approach for avoiding malignant glioma recurrence.</p>

Expression and clinical significance of HBsAg and HBcAg in hepatocytes in chronic hepatitis B / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the expression of HBsAg and HBcAg in hepatocytes in CHB patients, and analyze the correlation among the expression of HBsAg and HBcAg, the quantity of HBV DNA in serum, the pathology of liver tissue and the clinical manifestation.</p><p><b>METHODS</b>Quantitative polymerase chain reaction was used to assay the quantity of HBV DNA in serum in 351 CHB patients. Furthermore pathological diagnosis was performed using liver biopsy to assay the expression of HBsAg and HBcAg in hepatocytes by an immunohistochemical staining technique.</p><p><b>RESULTS</b>The positive expression rate of HBsAg and HBcAg in hepatocytes was 92.3% and 76.9% respectively. Cytoplasm-membrane HBcAg expression type (75.6%) was observed in the CHB with more active inflammation, while Nucleus HBcAg expression type (24.4%) was observed in the CHB with more sedative one (P < 0.0001). The expression of HBsAg was correlated with the quantity of HBV DNA in serum (rp = 0.24, P = 0.0129), while inversely correlated with the inflammation and the fibrillation of liver tissue (rp = -0.22, P = 0.0279; rp = -0.23, P = 0.0186). The expression of HBcAg was correlated with the quantity of HBV DNA in serum (rp = 0.52, P < 0.0001), while was inversely correlated with the inflammation and the fibrosis of liver (rp = -0.33, P < 0.0001; rp = -0.34, P < 0.0001).</p><p><b>CONCLUSION</b>Cytoplasm-membrane HBcAg expression type was observed in the CHB with more active inflammation, while Nucleus HBcAg expression type was observed in the CHB with mild change. In the immunopathogenesis of the liver damage in CHB, HBcAg might be a main target antigen. HBsAg might be a sensitive index to screen HBV infection; HBcAg might probably be a reliable index to evaluate the replication of HBV</p>

The value of gamma-glutamyltransferase in the diagnosis of chronic hepatitis B / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To explore the change of serum gamma-glutamyltransferase (GGT) and its diagnosis value in chronic hepatitis B (CHB) patients with different degrees of liver damage.</p><p><b>METHODS</b>Alanine-aminotransferase (ALT), aspartate-aminotransferase (AST) and GGT were measured in 221 CHB patients. Liver biopsy was conducted simultaneously to determine the inflammation grade and fibrosis stage of the liver tissues.</p><p><b>RESULTS</b>The rate of normal GGT in pathologically diagnosed mild and severe CHB patients was 90.4% and 12.3%, respectively (P<0.01). Increased level of GGT was parallel to the degree of liver pathological change (P<0.01). In active CHB patients, GGT rose with the ALT increase with a positive linear correlation between them (r=0.464, P<0.001). In pathologically diagnosed mild CHB patients, GGT had a tendency of rapidly declining to normal levels with ALT. In moderate CHB patients, GGT fluctuated at a relatively high level, and in severe CHB patients GGT exhibited a deviation from GGT.</p><p><b>CONCLUSIONS</b>GGT is conducive to improve the coincident rate between the clinical and pathological diagnosis of CHB.</p>

Evaluation of the value of quantitative assay of hepatitis B e antigen in diagnosis and therapy of chronic hepatitis B / 中华传染病杂志

Objective To evaluate the value of Microparticle Enzyme Immunoassay(MEIA),a quantitative assay, in the diagnosis and treatment of chronic hepatitis B (CHB). Methods MEIA, Enzyme linked immunoabsorbent assay (ELISA), and quantitative PCR were used to detect e antigen, hepatitis B serum markers, and the quantity of HBV DNA in 249 CHB patients and 32 non CHB patients respectively. Expression of HBcAg in Liver tissue was detected by using S P method. These measures were used to illuminate the relationships among serum e antigen quantity, hepatitis B serum markers, the quantity of HBV DNA, the quantity of HBcAg in hepatocytes, and pathologic diagnosis of liver tissues. Results 1. The sensitivity of MEIA (80.28%) to detect serum e antigen is higher than that of ELISA (69.01%)( ? 2=9.312, P =0.002).2. The serum e antigen quantity is positively correlated with the logarithm of the quantity of HBV DNA ( r =0.411, P =0.000). 3. The level of serum e antigen is positively correlated with the semi quantitative of HBcAg in hepatocytes ( r =0.646, P =0.000). 4. The serum e antigen quantity is negatively correlated with pathologic degree of liver tissues ( r =-0.172, P =0.006). Conclusions MEIA is a sensitive, stable, and reliable method to assay the serum e antigen quantity which could be used to evaluate the replication of HBV and the degree of liver damage.