Spider angiomas in Patients with liver Cirrhosis:Role of vascular endothelial growth factor and basic fibrobalast growth factor

AIM: To investigate whether vascular endothelial growth factor (VEGF) and basic fibroblastic growth factor (bFGF) are associated with spider angiomas in patients with liver cirrhosis.METHODS: Eighty-six patients with liver cirrhosis were enrolled and the number and size of the spider angiomas were recorded. Fifty-three healthy subjects were selected as controls. Plasma levels of VEGF and bFGF were measured in both the cirrhotics and the controls.RESULTS: Plasma VEGF and bFGF were increased in cirrhotics compared with controls (L22±13 vs. 71±11 pg/mL, P=0.003for VEGF; 5.1±0.5 vs. 3.4-±0.5 pg/mL, P=0.022 for bFGF). In cirrhotics, plasma VEGF and bFGF were also higher in patients with spider angiomas compared with patients without spider angiomas (185±28 vs. 90±10 pg/mL, P=0.003 for VEGF;6.8±1.0 vs. 4.1±0.5 pg/mL, P=0.017 for bFGF). Multivariate logistic regression showed that young age and increased plasma levels of VEGF and bFGF were the most significant predictors for the presence of spider angiomas in cirrhotic patients (odds ratio [OR]=6.64, 95 % confidence interval [CI]=2.02-21.79, P=0.002; OR=4.35, 95 % CI=1.35-14.01,P=0.014; OR=5.66, 95 % CI=1.72-18.63, P=0.004, respectively).CONCLUSION: Plasma VEGF and bFGF are elevated in patients with liver cirrhosis. Age as well as plasma levels of VEGF and bFGF are significant predictors for spider angiomas in cirrhotic patients.     

Expression of plasma vascular endothelial growth factor in patients with hepatocellular carcinoma and effect of transcatheter arterial chemoembolization therapy on plasma vascular endothelial growth factor level

AIM:To investigate the expression level of plasma vascularendothelial growth factor(P-VEGF)in patients withhepatocellular carcinoma(HCC)and its relationship withthe clinicopathologic characteristics,and to examine thechanges of P-VEGF in the course of transcatheter arterialchemoembolization(TACE).METHODS:Peripheral blood samples were taken from 45HCC patients before and 1,3,7 d,and 1 mo after TACE.Plasma VEGF level was measured with the quantitativesandwich enzyme-linked immunosorbent assay(ELISA).Twenty patients with benign liver lesions and 17 healthycontrol subjects were also included in this study.RESULTS:Plasma VEGF levels in HCC patients weresignificantly elevated as compared to those in patients withbenign liver lesions(P=0.006)and in the normal controls(P=0.003).Significant differences were observed whenP-VEGF was categorized by tumor size(P=0.006),portalvein thrombosis(P=0.011),distant metastasis(P=0.017),arterial-portal vein shunting(P=0.026),and InternationalUnion Against Cancer(UICC)TNM stage(P=0.044).Therewas no correlation between plasma level of VEGF and thelevel of alpha fetoprotein(α-FP) (r=0.068,P=0.658)andweakly correlated with the number of platelets(r=0.312,P=0.038).P-VEGF levels increased significantly andreached the peak value on the first day after TACE,and thendecreased gradually.The change rate of P-VEGF concentration(one month post-TACE/pre-TACEx100%)was correlatedwith the retention rate of lipiodol oil(rs=0.494,P=0.001)and the tumor volume change(rs=0.340,P=0.034).The patients who achieved a partial or complete responseto TACE therapy showed significantly less pre-treatmentP-VEGF than those nonresponders(P=0.025).A high pre-therapeutic P-VEGF level was associated with poor responseto treatment(P=0.018).CONCLUSION:A high pre-treatment P-VEGF level is auseful marker for tumor progression,especially for vascular invasion.TACE increases the level of P-VEGF onlytemporarily which may be associated with tumor ischemia.P-VEGF may be useful in predicting treatment response,monitoring disease course after TACE and judging the effectof different TACE regimens.     

Association of CT perfusion imaging with plasma levels of TGF-β1 and VEGF in patients with NSCLC

Objective: To study the association of CT perfusion imaging parameters with plasma level of transforming growth factor-β1(TGF-β1) and vascular endothelial growth(VEGF) in patients with non small cell cancer(NSCLC). Methods: A total of 67 patients with NSCLC(NSCLC group) and 64 patients with benign lesion(control group) were given with CT perfusion imaging to obtain blood flow, blood volume, mean transit time, time to peal and permeability surface through CT perfusion software. The plasma levels of TGF-β1 and VEGF were tested by ELISA. The relationship between plasma levels of TGF-β1, VEGF and CT perfusion imaging parameters were analyzed. Results: CT perfusion imaging parameters and the plasma levels of TGF-β1 and VEGF of NSCLC group were significantly higher than the control group(P0.05), while CT perfusion parameters and the levels of TGF-β1 and VEGF in NSCLC group showed significant difference in different tumor node metastasis stages(P0.05). Correlation analysis showed that the level of plasma TGF-β1 and VEGF were positively correlated with blood flow, blood volume, and mean transit time(P0.05), and negatively correlated with time to peal(P0.05). There was no significant correlation between TGF-β1 and VEGF with the permeability surface. Conclusions: CT perfusion imaging parameters in patients with NSCLC is closely associated with plasma TGF-β1, VEGF and its biological characteristics. CT perfusion imaging is a convenient method to detect tumor blood perfusion.     

Effect of lamivudine treatment on plasma levels of transforming growth factor β1, tissue inhibitor of metalloproteinases-1 and metalloproteinase-1 in patients with chronic hepatitis B

AIM: Transforming growth factor (TGF)-β1, metalloproteinase (MMP)-1 and its tissue inhibitor (TIMP)-I are considered predictive biomarkers of chronic hepatitis activity and fibrosis.The aim of this study was to evaluate the effect of lamivudine treatment on the plasma levels of these peptides in patients with chronic hepatitis B.METHODS: TGF-β1, MMP-1 and TIMP-1 plasma concentrations were measured with an enzyme immunoassay in 40 patients treated with lamivudine for 48 wk. Elimination of HBV-DNA and HBV antigens was evaluated 24 wk after treatment completion.RESULTS: Baseline TGF-β1(29.6±2.2 ng/mL) and TIMP-1(1 578±93 ng/mL) significantly exceeded normal values(18.3±1.6 ng/mL and 1 102±67 ng/mL respectively). Lamivudine treatment resulted in a significant decrease of TGF-β1 and TIMP-1 during treatment with an increase after 24 wk of treatment. Pretreatment MMP-1 levels (6.7±0.7 ng/mL) were significantly lower than normal values (11.9±0.9 ng/mL) and increased during treatment and follow-up. A significant correlation was noted between TGF-β1 or TIMP-1 and aminotransferases as well as fibrosis scored in liver biopsy specimens. There were no statistically significant differences of TGF-β1, TIMP-1 and MMP-1 between four groups at baseline, 24 and 48 wk of treatment. TGF-β1 and TIMP-1 levels increased significantly in non-responders and normalized in responders at wk 72. MMP-1 also normalized in responders and decreased to values significantly lower than normal in non-responders.CONCLUSION: These findings support the role of TGF-β1,TIMP-1 and MMP-1 in the pathogenesis of chronic hepatitis B.Because of their association with hepatic injury and antiviral treatment efficacy, determination of these peptides may be useful in disease management.     

Angiogenic synergistic effect of basic fibroblast growth factor and vascular endothelial growth factor in an in vitro quantitative microcarrier-based three-dimensional fibrin angiogenesis system

AIM:To develop an in vitro three-dimensional(3-D)angiogenesis system to analyse the capillary sprouts inducedin response to the concentration ranges of basic fibroblastgrowth factor(bFGF)and vascular endothelial growth factor(VEGF)and to quantify their synergistic activity.METHODS:Microcarriers(MCs)coated with humanmicrovascular endothelial cells(HMVECs)were embeddedin fibrin gel and cultured in 24-well plates with assaymedia.The growth factors bFGF,or VEGF,or both wereadded to the system.The wells(n=8/group)were digitallyphotographed and the average length of capillary-like sprouts(ALS)from each microcarrier was quantitated.RESULTS:In aprotinin-stabilized fibrin matrix,humanmicrovascular endothelial cells on the MCs invaded fibrin,forming sprouts and capillary networks with lumina.Theangiogenic effects of bFGF or VEGF were dose-dependent inthe range from 10 to 40 ng/mL.At d 1,10 ng/mL of bFGF andVEGF induced angiogenesis with an ALS of 32.13±16.6 μmand 43.75:1:27.92 IJm,respectively,which were significantlyhigher than that of the control(5.88±4.45 μm,P<0.01),and the differences became more significant as the timeincreased.In addition,the combination of 20 ng/mL ofbFGF and VEGF each induced a more significant effect thanthe summed effects of bFGF(20 ng/mL)alone and VEGF(10 ng/mL)alone when analyzed using SPSS system forgeneral linear model(GLM)(P= 0.011),and that also exceededthe effects by 20 ng/mL of either bFGF or VEGF.CONCLUSION:A microcarrier-based in vitro three-dimensional angiogenesis model can be developed in fbrin.It offers a unique system for quantitative analysis ofangiogenesis.Both bFGF and VEGF exert their angiogeniceffects on HMVECs synergistically and in a dose-dependentmanner.     

Effect of pegylated interferon alpha 2b plus ribavirin treatment on plasma transforming growth factor-β1,metalloproteinase-1,and tissue metalloproteinase inhibitor-1 in patients with chronic hepatitis C

AIM: To evaluate the effect of antiviral treatment on plasma levels of transforming growth factor-β1 (TGF-β1), metalloproteinase 1 (MMP-1), and tissue inhibitor of metalloproteinase-1 (TIMP-1) in patients with chronic hepatitis C. METHODS: TGF-β1, MMP-1, and TIMP-1 plasma concentrations were measured by an enzyme immunoassay in 28 patients, during 48 wk of treatment with pegylated interferon-alpha 2b (PEG-IFN-α2b) plus ribavirin (RBV) and after 24 wk of follow-up. Patients were divided into two groups: responders (R) and non-responders (NR) related to achieved sustained virologic response. Normal values were evaluated in plasma samples of 13 healthy volunteers. RESULTS: Baseline plasma concentrations of TGF-βl and TIMP-1 (30.9±3.7 and 1 506±61 ng/mL respectively) measured in all subjects significantly exceeded the normal values (TGF-β1: 18.3±1.6 ng/mL and TIMP-1: 1 102±67 ng/mL). In contrast, pretreatment MMP-1 mean level (6.5±0.9 ng/mL) was significantly lower than normal values (11.9±0.9 ng/mL). Response to the treatment was observed in 12 patients (43%). TGF-β1 mean concentration measured during the treatment phase decreased to the control level in both groups. However at wk 72, values of NR patients increased and became significantly higher than in R group. TIMP-1 concentrations in R group decreased during the treatment to the level similar to normal. In NR group, TIMP-1 remained significantly elevated during treatment and follow-up phase and significant difference between both groups was demonstrated at wk 48 and 72. MMP-1 levels were significantly decreased in both groups at baseline. Treatment caused rise of its concentration only in the R group, whereas values in NR group remained on the level similar to baseline. Statistically significant difference between groups was noted at wk 48 and 72. CONCLUSION: These findings support the usefulness of TGF-β1, TIMP-1, and MMP-1 in the management of chronic hepatitis C. Elevated TIMP-1 and low MMP-1 plasma concentrations during antiviral therapy may indicate medication failure.     

Diagnostic value of platelet derived growth factor-BB, transforming growth factor-β1,matrix metalloproteinase-1, and tissue inhibitor of matrix metaUoproteinase-1 in serum and peripheral blood mononuclear cells for hepatic fibrosis

AIM: Noninvasive diagnosis of hepatic fibrosis has become the focus because of the limited biopsy, especially in the surveillance of treatment and in screening hepatic fibrosis. Recently, regulatory elements involved in liver fibrosis, such as platelet derived growth factor-BB (PDGF-BB), transforming growth factor-β1(TGF-β1), matrix metalloproteinase-1 (MMP-1), and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1), have been studied extensively. To determine whether these factors or enzymes could be used as the indices for the diagnosis of hepatic fibrosis, we investigated them by means of receiver operating characteristic (ROC) curve. METHODS: Serum samples from sixty patients with chronic viral hepatitis B and twenty healthy blood donors were assayed to determine the level of PDGF-BB, TGF-β1, MMP-1, and TIMP-1 with ELISA, and HA, PCIII, C-IV, and LNlevel with RIA. The message RNA (mRNA) expression of TIMP-1 and MMP-1 in peripheral blood mononuclear cells (PBMCs) was detected by RT-PCR and Northern blot hybridization. Liver biopsy was performed in all patients. The biopsy samples were histopatholocjically examined. The trial was double-blind controlled. RESULTS: The serum level of PDGF-BB, TIMP-1, the ratio of TIMP-1 and MMP-1 (TIMP-1/MMP-1), mRNA expression of TIMP-1 (TIMP-lmRNA), and the ratio of TIMP-lmRNA and MMP-lmRNA (TIMP-lmRNA/MMP-lmRNA) in patients was significantly higher than those in the healthy blood donors (t=2.514-11.435, P=0.000-0.016). The serum level of PDGF-BB, TIMP-1, TIMP-1/MMP-1, and TIMP-lmRNA was positively correlated with fibrosis stage and inflammation grade (r=0.239-0.565, P=0.000-0.033), while the serum level of MMP-1 was negatively correlated with fibrosis stage and inflammation grade, and TIMP-lmRNA/MMP-lmRNA was positively correlated with inflammation grade. Through the analysis by ROC curve, serum PDGF-BB was the most valuable marker, and its sensitivity was the highest among the nine indices. The markers with the highest specificity were TIMP-lmRNA and TIMP-lmRNA/MMP-lmRNA in PBMCs. The area under the curve (AUC) of PDGF-BB, TIMP-lmRNA, TIMP-lmRNA/MMP-lmRNA, TIMP-1/MMP-1, HA, PCIII, TIMP-1, C-IV, and LN was 0.985, 0.876, 0.792, 0.748, 0.728, 0.727, 0.726, 0.583, and 0.463, respectively. The sensitivity and the specificity in the parallel test was 99.0% and 95.0 % when serum PDGF-BB, TIMP-lmRNA and TIMP-lmRNA/MMP-lmRNA was detected simultaneously. CONCLUSION: Serum level of PDGF-BB, TIIVIP-lmRNA, TIMP-lmRNA/MMP-lmRNA in PBMCs, and serum level of TIMP-1 and TIMP-1/MMP-1 can be used as the indices for the diagnosis of hepatic fibrosis, but the former three are more useful. The combination of serum PDGF-BB, TIMP-lmRNA and TIMP-lmRNA/MMP-lmRNA in PBMCs is even more efficient in screening liver fibrosis.     

Decreased plasma urotensin Ⅱ levels inversely correlate with extent and severity of coronary artery disease

Objective To determine the plasma urolensin Ⅱ(UII) levels in various types of coronary heart disease and to clarify how the plasma UII levels correlate with the clinical presentation, extent and severity of coronary artery atherosclerosis (CAD). Methods: One hundred and three aged patients undergoing elective diagnostic coronary angiography for proven or clinical suspected coronary heart disease were enrolled in this study. The extent and severity of coronary artery disease were evaluated by vessel score and Gensini score, respectively. Plasma UII levels were measured by radioimmunoassay. Results: The plasma UII levels in the patients with modest to severe coronary stenosis (3.03±0.34 pg/ml, 1.83±0.67 pg/ml) were significantly lower than that in subjects with normal coronary artery (4.80±1.11 pg/ml, P<0.001). The plasma UII levels in patients with coronary heart disease were also significantly lower than that in patients with insignificant coronary stenosis (P < 0. 001). Compared to patients with stable angina pectoris, plasma UII levels in patients with acute coronary syndrome were significantly decreased (1.89±0.51 pg/ml vs 2.42±0.77 pg/ml, P < 0.001). Plasma UII levels were found to be negatively correlated with the severity of coronary artery stenosis (r = -0.488, P<0.001), as well as the vessel score (r = -0.408, P<0.05) in the patients with CAD. Conclusion: Significant inverse correlations exist between the plasma UII levels, and the extent and severity of coronary artery stenosis. These findings suggest that plasma UII contribute to the development and progression of coronary artery stenosis, and may be a novel marker to predict clinical types, as well as the extent and severity of coronary artery disease in the patients.     

Relationship between serum cytokine levels and histopathological changes of liver in patients with hepatitis B

AIM: To investigate whether there was a relationship between the liver functions and fibrosis scores of hepatitis B patients and their TNF-α, IFN-γ,IL-4, and TGF-β1 serum levels based on the studies of liver biopsies. METHODS: Thirty patients with chronic hepatitis B (CHB) receiving no treatment and 30 healthy individuals with negative hepatitis serology and normal values of liver biochemistry were studied. After serum samples of the patients were collected, liver needle biopsy was performed on each patient. Cytokine levels were studied by ELISA. The biopsy materials were scored based on Knodell's histological activity index. RESULTS: In comparison of cytokine levels between CHB patients and control group, TNF-α,IL-4, and TGPβ1 levels of the patients were higher in CHB patients than in the controls, while IFN-γ level was lower in the patients than in the controls. There were significant differences between the groups in TNF-α, IL-4, TGF-β1, and IFN-γ(P<0.005, 0.03, 0.002, 0.0001,respectively).There was a negative correlation between TGF-β1 and IL-4 and IFN-γ(P<0.05), TNF-α and the other cytokines and IFN-γ and IL-4 were not correlated (P>0.05). TGF-β1 was correlated with fibrosis (P<0.05).Liver necroinflammatory activity and fibrosis and TNF-α, IL-4, and IFN-γ were not correlated (P>0.05). CONCLUSION: In the course of HBV infection and its chronic progress, cytokines play an important role. IL-4 and IFN-γ are effective in the chronic progression, while TGF-β1 is effective in the development of fibrosis. Serum cytokine levels may be effective tools in the estimation of chronic progression and fibrosis development.     

Effect of lamivudine treatment on plasma levels of transforming growth factor β_1,tissue inhibitor of metalloproteinases-1 and metalloproteinase-1 in patients with chronic hepatitis B

AIM:Transforming growth factor (TGF)-β_1,metalloproteinase(MMP)-1 and its tissue inhibitor (TIMP)-1 are consideredpredictive biomarkers of chronic hepatitis activity and fibrosis.The aim of this study was to evaluate the effect of lamivudinetreatment on the plasma levels of these peptides in patientswith chronic hepatitis B.METHODS:TGF-β_1,MMP-1 and TIMP-1 plasma concentrationswere measured with an enzyme immunoassay in 40 patientstreated with lamivudine for 48 wk.Elimination of HBV-DNAand HBV antigens was evaluated 24 wk after treatmentcompletion.RESULTS:Baseline TGF-β (29.6±2.2 ng/mL) and TIMP-1(1 578±93 ng/mL) significantly exceeded normal values(18.3±1.6 ng/mL and 1 102±67 ng/mL respectively).Lamivudinetreatment resulted in a significant decrease of TGF-β_1 andTIMP-1 during treatment with an increase after 24 wk oftreatment.Pretreatment MMP-1 levels (6.7±0.7 ng/mL) weresignificantly lower than normal values (11.9±0.9 ng/mL) andincreased during treatment and follow-up.A significantcorrelation was noted between TGF-β_1 or TIMP-1 andaminotransferases as well as fibrosis scored in liver biopsyspecimens.There were no statistically significant differencesof TGF-β_1,TIMP-1 and MMP-1 between four groups atbaseline,24 and 48 wk of treatment.TGF-β_1 and TIMP-1levels increased significantly in non-responders and normalizedin responders at wk 72.MMP-1 also normalized in respondersand decreased to values significantly lower than normal innon-responders.CONCLUSION:These findings support the role of TGF-β_1,TIMP-1 and MMP-1 in the pathogenesis of chronic hepatitis B.Because of their association with hepatic injury and antiviraltreatment efficacy,determination of these peptides may beuseful in disease management.     

C-reactive protein,procalcitonin,interleukin-6,vascular endothelial growth factor and oxidative metabolites in diagnosis of infection and staging in patients with gastric cancer

AIM:The current study was to determine the serum/pLasmalevels of VEGF,IL-6,malondialdehyde (MDA),nitric oxide(NO),PCT and CRP in gastric carcinoma and correlation withthe stages of the disease and accompanying infection.METHODS:We examined the levels of serum VEGF,IL-6,PCT,CRP and plasma MDA,NO in 42 preoperative gastriccancer patients and 23 healthy subjects.There were infectionanamneses that had no definite origin in 19 cancer patients.RESULTS:The VEGF levels (mean±SD; pg/mL) were478.05±178.29 and 473.85±131.24 in gastric cancer patientswith and without infection,respectively,and these valueswere not significantly different (P>0.05).The levels of VEGF,CRP,PCT,It-6,MDA and NO in cancer patients weresignificantly higher than those in healthy controls and thelevels of CRP,PCT,It-6,MDA and NO were statisticallyincreased in infection group when compared with non-infection group (P<0.001).CONCLUSION:Although serum VEGF concentrations wereincreased in gastric cancer,this increase might not be relatedto infection.CRP,PCT,IL-6,MDA and NO have obviousdrawbacks in the diagnosis of infections in cancer patients.These markers may not help to identify infections in theprimary evaluation of cancer patients and hence to avoidunnecessary antibiotic treatments as well as hospitalization.According to the results of this study,IL-6,MDA,NO andespecially VEGF can be used as useful parameters todiagnose and grade gastric cancer.     

Helicobacter pylori eradication lowers serum homocysteine level in patients without gastric atrophy

AIM: To determine whether Helicobacter pylori (H pylori) infection caused hyperhomocysteinemia by altering serum vitamin B_(12), serum folate and erythrocyte folate levels and whether eradication of this organism decreased serum homocysteine level. METHODS: The study involved 73 dyspeptic H pylork positive patients, none of them had gastric mucosal atrophy based on rapid urease test and histology. Out of 73 patients, 41 (56.2%) showed a successful eradication of H pylori 4 wk after the end of treatment. In these 41 patients, fasting serum vitamin B_(12) folate and homocysteine levels, and erythrocyte folate levels before and 4 wk after H pylori eradication therapy were compared. RESULTS: The group with a successful eradication of H pylori had significantly higher serum vitamin B_(12) and erythrocyte folate levels in the post-treatment period compared to those in pre-treatment period (210±97 pg/mL vs 237±94 pg/mL,P＜0.001 and 442±212 ng/mL vs 539±304 ng/mL, P=0.024, respectively), but showed no significant change in serum folate levels (5.6±2.6 ng/mL vs 6.0+2.4 ng/mL, P=0.341). Also, the serum homocysteine levels in this group were significantly lower after therapy (13.1±5.2 μmol/L vs 11.9±6.2 μmol/L, P=0.002). Regression analysis showed that serum homocysteine level was positively correlated with age (P=0.01) and negatively with serum folate level before therapy (P=0.003). CONCLUSION: Eradication of H pylori decreases serum homocysteine even in patients who do not exhibit gastric mucosal atrophy. It appears that the level of homocysteine in serum is related to a complex interaction among serum vitamin B_(12), serum folate and erythrocyte folate levels.     

Effect of lamivudinein in BeAg-positive chronic hepatitis B: Discordant effect on HBeAg and HBV DNA according to pretreatment ALT level

AM: To clarify differences in antiviral effect of the drug in patients with different ALT levels, we examined the changes in HBV markers in patients with high or low ALT levels with or without lamivudine treatment. METHODS: Thirty-seven HBeAg-positive patients were studied. Ten patients with ALT levels higher than 200 IU/L (group 1) and 8 patients with ALT below 200 IU/L (group 2) were treated orally with 100 mg/d of lamivudine. As untreated control, 9 patients with ALT above 200 IU/L (group 3) and 10 patients with ALT below 200 IU/L (group 4) were examined. ALT level, HBeAg/HBeAb status, and HBV DNA level were examined monthly for 11.9±0.4 mo. RESULTS: The ALT level normalized in all 10 patients of group 1, 7/8 of group 2, 4/9 of group 3, and 1/10 of group 4 within 6 mo (groups 1 vs2, P= NS; groups 1 vs 3, P= 0.002; groups 1 vs4, P<0.0001). HBV DNA fell below the detection limit in all 10 patients of group 1, 7/8 of group 2, 0/9 of group 3, and 0/10 of group 4 within 6 mo (groups 1 vs 2, P - NS). HBeAg became seronegative in 7/10 patients of group 1, 1/8 of group 2, 3/9 of group 3, and 0/10 of group 4 within 12 mo (groups 1 vs2, P= 0.02; groups 1 vs 3, P= NS). CONCLUSION: Our data suggest that HBeAg-positive patients with higher ALT levels can be considered good candidates for lamivudine therapy, probably because lamivudine accelerates the natural seroconversion of HBeAg, accompanied by HBV DNA loss, in these patients.     

Correlation between anti-fibrotic effect of baicalin and serum cytokines in rat hepatic fibrosis

AIM： To investigate the correlation between the antifibrotic effect of baicalin and serum cytokine production in rat hepatic fibrosis, METHODS： Forty male Sprague-Dawley rats were divided randomly into four groups： normal control group, model group, baicalin-treated group, and colchicine-treated group. Except for the normal control group, all rats in the other groups were administered with carbon tetrachloride to induce hepatic fibrosis. At the same time, the last two groups were also treated with baicalin or colchicine. At the end of the 8 wk, all animals were sacrificed. Serum alanine aminotransferase （ALl＇）, aspartate aminotransferase （AST）, transforming growth factor （TGF）-β1, tumor necrosis factor （TNF）-α, interleukin （IL）-6 and IL-10 were measured. Liver index, hepatic hydroxyproline content and the degree of liver fibrosis were also evaluated. RESULTS： The levels of ALT, AST and liver index in the baicalin-treated group were markedly lower than those in the model group （ALT： 143.88 ± 14.55 U/L vs 193.58± 24.35 U/L; AST： 263.66 ± 44.23 U/L vs 404.37± 68.29 U/L; liver index： 0.033 ± 0.005 vs 0.049± 0.009, P 〈 0.01）. Baicalin therapy also significantly attenuated the degree of hepatic fibrosis, collagen area and collagen area percentage in liver tissue （P 〈 0.01）. Furthermore, the levels of serum TGF-β1, TNF-α and IL-6 were strikingly reduced in the baicalin-treated group compared with the model group, while the production of IL-10 was up-regulated： （TGF-β1：260.21 ± 31.01 pg/mL vs 375.49 ± 57.47 pg/mL; TNF-α： 193.40±15.18 pg/mL vs 260.04 ± 37.70 pg/mL; IL-α：339.87 ± 72.95 pg/mL vs 606.47 ± 130.73 pg/mL; IL-10：506.22 ± 112.07 pg/mL vs 316.95 ± 62.74 pg/mL, P 〈 0.01）. CONCLUSION： Baicalin shows certain therapeutic effects on hepatic fibrosis, probably by immunoregulating the imbalance between profibrotic and antifibrotic cytokines.     

Regulatory polymorphism of CXCL10 rs1439490 in seronegative occult hepatitis C virus infection

AIM To examine the relationship between the single nucleotide polymorphism CXCL10 rs1439490 and seronegative occult hepatitis C virus(HCV) infection(OCI).METHODS One hundred and three cases of seronegative OCI and 155 cases of seropositive chronic HCV infection(CHC) were diagnosed at five Liver Centers in Northeastern China, from 2012 to 2016. CXCL10 rs1439490, rs1440802, and IL-28 B rs12979860 were analyzed by sequencing. Serum CXCL10 was measured by ELISA. Intrahepatic CXCL10 was determined by quantitative PCR and immunohistochemical semi-quantitative scoring. Liver necroinflammation and fibrosis were scored according to the METAVIR system.RESULTS CXCL10 rs1439490 G/G was more prevalent in OCI patients(n = 93/103; 90.3%) than in CHC patients(n = 116/155; 74.8%; P = 0.008). OCI patients had lower serum CXCL10 levels than CHC patients(192.91 ± 46.50 pg/mL vs 354.78 ± 102.91 pg/mL, P 0.0001). Of IL-28 B rs12979860 C/C patients, OCI patients with rs1439490 G/G had lower serum and liver levels of CXCL10 and lower levels of liver necroinflammation and fibrosis than non-G/G patients. OCI patients had higher alanine aminotransferase normalization rates after Peginterferon treatment than CHC patients(P 0.05) and serum CXCL10 decreased significantly(P 0.0001). Liver necroinflammation and fibrosis were alleviated in 8 OCI patients after treatment. Multivariate analysis indicated that rs1439490 G/G significantly influenced the occurrence of OCI in HCV infection(OR = 0.31, 95%CI: 0.15-0.66, P = 0.002).CONCLUSION CXCL10 rs1439490 G/G is positively associated with OCI in HCV infection and antiviral outcome.     

Blockage of transforming growth factor β receptors prevents progression of pig serum-induced rat liver fibrosis

AIM: To test the hypothesis that introduction of antisense TβR Ⅰ and TβR Ⅱ eukaryotic expressing plasmids into a rat model of immunologically induced liver fibrosis might block the action of TGF-β1 and halt the progression of liver fibrosis. METHODS: RT-Nest-PCR and gene recombination techniques were used to construct rat antisense TβR Ⅰ and TβR Ⅱ recombinant plasmids which could be expressed in eukaryotic cells. The recombinant plasmids and empty vector (pcDNA3) were encapsulated by glycosyl-poly-L-lysine and then transducted into rats of pig serum-induced liver fibrosis model. Expression of exogenously transfected gene was assessed by Northern blot, and hepatic expressions of TβR Ⅰ and TβR Ⅱ were evaluated by RT-PCR and Western blot.We also performed ELISA for serum TGF-β1, hydroxyproline of hepatic tissues, immunohistochemistry for collagen types Ⅰ and Ⅲ, and VG staining for pathological study of the liver tissues. RESULTS: The exogenous antisense TβR Ⅰ and TβR Ⅱ plasmids could be well expressed in vivo, and block mRNA and protein expression of TβR Ⅰ and TβR Ⅱ in the fibrotic liver at the level of mRNA respectively. These exogenous plasmid expressions reduced the level of TGF-β1 (antisense TβR Ⅰ group 23.998+3.045 ng/mL, antisense TβR Ⅱ group 23.156+3.131 ng/mL, disease control group 32.960+3.789 ng/mL; F-=-38.19, 36.73, P&lt;0.01). Compared with disease control group, the contents of hepatic hydroxyproline (antisense TβR Ⅰ group 0.169+0.015 mg/g liver, antisense TβR Ⅱ group 0.167+0.009 mg/g liver, disease control group 0.296+0.026 mg/g liver; F=14.39, 15.48, P&lt;0.01) and the deposition of collagen types Ⅰ and Ⅲ decreased in the two antisense treatment groups(antisense TβR Ⅰ group, collagen type Ⅰ 669.90+50.67, collagen type Ⅲ 657.29+49.48; antisense TβR Ⅱ group, collagen type Ⅰ 650.26+51.51, collagen type Ⅲ 661.58+55.28; disease control group, collagen type I 1209.44+116.60, collagen type Ⅲ 1175.14+121.44; F=15.48 to 74.89, P&lt;0.01). Their expression also improved the pathologic classification of liver fibrosis models (compared with disease control group, X^2=17.14, 17.24, P&lt;0.01). No difference was found in the level of TGF-β1, the contents of hepatic hydroxyproline and collagen types Ⅰ and Ⅲ and pathologic grade between pcDNA3 control group and disease control group or between the two antisense treatment groups (F=0.11 to1.06, X^2=0.13 to 0.16, P&gt;0.05). CONCLUSION: Antisense TβR Ⅰ and TβR Ⅱ recombinant plasmids have certain reverse effects on liver fibrosis and can be used as possible candidates for gene therapy.     

Increased osteopontin and liver stiffness measurement by transient elastography in biliary atresia

AIM: To analyze plasma osteopontin levels and liver stiffness using transient elastography in postoperative biliary atresia (BA) children compared with healthy controls. METHODS: Thirty children with postoperative BA and 10 normal controls were enrolled. The patients were categorized into two groups according to their jaundicestatus. Plasma levels of osteopontin were determined using commercially available enzyme-linked immunosorbent assay. Liver stiffness was measured by using transient elastography (Fibroscan). Ten validated Fibroscan measurements were performed in each patient and control with the result expressed in kilopascals (kPa). RESULTS: Plasma osteopontin was significantly elevated in BA children compared with that of healthy controls (47.0 ± 56.4 ng/mL vs 15.1 ± 15.0 ng/mL, P = 0.01). The liver stiffness measurement was markedly elevated in the patients with BA compared with that of controls (26.9 ± 24.6 kPa vs 3.9 ± 0.7 kPa, P = 0.001). Subgroup analysis showed that the BA patients with jaundice had more pronounced plasma osteopontin levels than those without jaundice (87.1 ± 61.6 ng/mL vs 11.9 ± 6.1 ng/mL, P = 0.001). Furthermore, the mean liver stiffness was significantly greater in the jaundiced BA patients compared with non-jaundiced patients (47.7 ± 21.8 kPa vs 8.7 ± 3.0 kPa, P = 0.001). Additionally, plasma osteopontin was positively related to serum total bilirubin (r = 0.64, P < 0.001). There was also a correlation between plasma osteopontin and liver stiffness values (r = 0.60, P < 0.001). CONCLUSION: High plasma osteopontin positively correlated with degree of hepatic fibrosis and could be used as a biochemical parameter reflecting disease severity in postoperative BA children.