白细胞介素-17肝内表达与慢性乙型肝炎病毒感染所致肝纤维化的相关性

Objective To explore the relationship between intrahepatic expression of interleukin-17 (IL-17) and liver fibrosis in patients with chronic hepatitis B virus infection. Methods IL-17 expressions in livers with different inflammation activity grades and hepatic fibrosis stages from patients with chronic hepatitis B virus carriers (n= 30), chronic hepatitis B (CHB, n = 55), liver cirrhosis (LC, n=20) were measured by immunohistochemistry. Serum IL-17 and liver fibrosis indices of haluronic acid (HA), laminin (LN), type Ⅲ procollagen (PC Ⅲ ) and type Ⅳ collagen ( Ⅳ C) were determined by enzyme linked immunosorbent assay (ELISA). The differences between groups were compared by Kruskal-Wallis test, and the Mann-Whitney test, and the correlation analysis was done by Spearman test. Results Intrahepatic IL-17 expression in LC group was significantly higher than CHB group (x2 =25. 3982, P=0. 004), and that in CHB group was higher than chronic hepatitis B virus carriers group (x2 = 11. 5056, P= 0. 001). The inflammation activity grade and hepatic fibrosis stage were both positively correlated with IL-17 expression (r= 0.718, 0. 693, respectively; both P＜0.01). IL-17 mainly located in portal area and the expression was positively correlated with serum levels of HA, LN, PCⅢ and ⅣC (r=0. 793, 0. 834, 0. 722, 0. 883, respectively; all P＜0.01).Conclusion Intrahepatic IL-17 expression is closely correlated with liver inflammation activity grade and hepatic fibrosis stage.     

丙氨酸转氨酶正常的慢性乙型肝炎病毒感染者的肝脏病理学分析

Objective To study the liver histological changes in chronic hepaitits B (CHB) patients with normal serum alanine aminotransferase (ALT) levels and the related factors. Methods Six hundred and thirty-two CHB patients with normal ALT levels had undergone ultrasound guided percutaneous liver biopsies. All specimen were examined by HE staining, collagen fiber Masson staining and immunohistochemical staining for hepatitis B surface antigen (HBsAg) and hepatitis B core antigen (HBcAg). The Knodell inflammation score and Ishak fibrosis score were both calculated and the relationship with age, serum levels of ALT and hepatitis B virus (HBV) DNA, hepatic expressions of HBsAg and HBcAg were analyzed. The means between two groups were compared by t test and those among groups were compared by one-factor analysis of variance and q test. Enumeration data were analyzed by x2 test. Results Among 632 CHB patients with normal ALT levels, 167 (26.4%) showed moderate necrotic inflammation in liver tissues and 26 (4.1%) showed severe necrotic inflammation; 217 (34. 3 % ) showed moderate fibrosis and 52 (8. 2 % ) showed severe fibrosis (cirrhosis). The Knodell inflammation score and Ishak fibrosis score in high ALT group were higher than low ALT group, those in female high ALT group were higher than male high ALT group and those in patients ＞ 40 years old were higher than ≤20 years old (q= 19.63, P＜0. 05). The liver injuries in patients with active HBV replication were more severe than those with undetectable HBV DNA levels (Knodell score, q=3.87, 2.87, 6.34; Ishak score, q=2.64,2. 64,5.54, all P＜0. 05),while there was no significant difference between patients with high levels and low levels of HBV DNA (F= 1.35, P＞0. 05). There was no significant difference between expressions of HBsAg (F= 1.65,0. 73,respectively; both P＞0. 05) and HBcAg in liver tissues and Knodell inflammation score and Ishak fibrosis score (F=0. 17, 1.29, respectively; both P＞0. 05). Conclusions Liver biopsies should be considered in CHB patients with normal ALT levels and detectable HBV DNA levels, especially those ＞ 40 years old and with ALT of (0.75-1.00) × upper limits of normal (ULN).     

丙氨酸转氨酶正常的慢性乙型肝炎病毒感染者的肝脏病理学分析

Objective To study the liver histological changes in chronic hepaitits B (CHB) patients with normal serum alanine aminotransferase (ALT) levels and the related factors. Methods Six hundred and thirty-two CHB patients with normal ALT levels had undergone ultrasound guided percutaneous liver biopsies. All specimen were examined by HE staining, collagen fiber Masson staining and immunohistochemical staining for hepatitis B surface antigen (HBsAg) and hepatitis B core antigen (HBcAg). The Knodell inflammation score and Ishak fibrosis score were both calculated and the relationship with age, serum levels of ALT and hepatitis B virus (HBV) DNA, hepatic expressions of HBsAg and HBcAg were analyzed. The means between two groups were compared by t test and those among groups were compared by one-factor analysis of variance and q test. Enumeration data were analyzed by x2 test. Results Among 632 CHB patients with normal ALT levels, 167 (26.4%) showed moderate necrotic inflammation in liver tissues and 26 (4.1%) showed severe necrotic inflammation; 217 (34. 3 % ) showed moderate fibrosis and 52 (8. 2 % ) showed severe fibrosis (cirrhosis). The Knodell inflammation score and Ishak fibrosis score in high ALT group were higher than low ALT group, those in female high ALT group were higher than male high ALT group and those in patients ＞ 40 years old were higher than ≤20 years old (q= 19.63, P＜0. 05). The liver injuries in patients with active HBV replication were more severe than those with undetectable HBV DNA levels (Knodell score, q=3.87, 2.87, 6.34; Ishak score, q=2.64,2. 64,5.54, all P＜0. 05),while there was no significant difference between patients with high levels and low levels of HBV DNA (F= 1.35, P＞0. 05). There was no significant difference between expressions of HBsAg (F= 1.65,0. 73,respectively; both P＞0. 05) and HBcAg in liver tissues and Knodell inflammation score and Ishak fibrosis score (F=0. 17, 1.29, respectively; both P＞0. 05). Conclusions Liver biopsies should be considered in CHB patients with normal ALT levels and detectable HBV DNA levels, especially those ＞ 40 years old and with ALT of (0.75-1.00) × upper limits of normal (ULN).     

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.     

Serum interleukin-34 level can be an indicator of liver fibrosis in patients with chronic hepatitis B virus infection

AIM To investigate whether serum interleukin(IL)-34 levels are correlated with hepatic inflammation and fibrosis in patients with chronic hepatitis B virus(HBV) infection. METHODS In this study, serum IL-34 levels were assessed by enzyme-linked immunosorbent assay in 19 healthy controls and 175 patients with chronic HBV infection undergoing biopsy. The frequently used serological markers of liver fibrosis were based on laboratory indexes measured at the Clinical Laboratory of the Second Affiliated Hospital of Anhui Medical University. Liver stiffness was detected by transient elastography with Fibro Touch. The relationships of non-invasive makers of liver fibrosis and IL-34 levels with inflammation and fibrosis were analyzed. The diagnostic value of IL-34 and other liver fibrosis makers wereevaluated using areas under the receiver operating characteristic curves, sensitivity and specificity.RESULTS Serum IL-34 levels were associated with inflammatory activity in the liver, and IL-34 levels differed among phases of chronic HBV infection(P = 0.001). By comparing serum IL-34 levels among patients with various stages of liver fibrosis determined by liver biopsy, we found that IL-34 levels ≥ 15.83 pg/m L had a high sensitivity of 86.6% and a specificity of 78.7% for identifying severe fibrosis(S3-S4). Furthermore, we showed that IL-34 is superior to the fibrosis-4 score, one of the serum makers of liver fibrosis, in identifying severe liver fibrosis and early cirrhosis in patients with HBV-related liver fibrosis in China.CONCLUSION Our results indicate that IL-34, a cytokine involved in the induction of activation of profibrogenic macrophages, can be an indicator of liver inflammation and fibrosis in patients with chronic HBV infection.     

Effects of six months Iosartan administration on liver fibrosis in chronic hepatitis C patients： A pilot study

AIM: To evaluate the safety and efficacy of chronic administration of losartan on hepatic fibrosis in chronic hepatitis C patients. METHODS: Fourteen patients with chronic hepatitis C non-responders (n = 10), with contraindications (n = 2) or lack of compliance (n = 2) to interferon plus ribavirin therapy and liver fibrosis were enrolled. Liver and renal function test, clinical evaluation, and liver biopsies were performed at baseline and after losartan administration at a dose of 50 mg/d during the 6 mo. The control group composed of nine patients with the same inclusion criteria and paired liver biopsies (interval 6-14 mo). Histological activity index (HAI) with fibrosis stage was assessed under blind conditions by means of Ishak's score. Subendothelial fibrosis was evaluated by digital image analyses. RESULTS: The changes in the fibrosis stage were significantly different between losartan group (decrease of 0.5±1.3) and controls (increase of 0.89±1.27; P<0.03). In the treated patients, a decrease in fibrosis stage was observed in 7/14 patients vs 1/9 control patients (P<0.04). A decrease in sub-endothelial fibrosis was observed in the losartan group. No differences were found in HAI after losartan administration. Acute and chronic decreases in systolic arterial pressures (P<0.05) were observed after the losartan administration, without changes in mean arterial pressure or renal function. CONCLUSION: Chronic AT-Ⅱ type 1 receptor (AT1R) blockade may reduce liver fibrosis in patients with chronic hepatitis C.     

Serum vitamin D and vitamin-D-binding protein levels in children with chronic hepatitis B

BACKGROUND Vitamin D is an essential fat-soluble secosteroid hydroxylated by the liver to form the intermediate metabolite calcidiol{25-hydroxy vitamin D[25(OH)D]},which is a reliable indicator to investigate individual vitamin D status.Vitamin-D-binding protein(VDBP)is a multifunctional glycoprotein mainly synthesized in the liver and the major transport protein for vitamin D and its metabolites.Serum vitamin D and VDBP are both associated with hepatitis B.However,few studies have reported the relationship and clinical significance of vitamin D and VDBP with hepatitis B virus(HBV)replication and hepatic fibrosis in children with chronic hepatitis B(CHB).AIM To explore vitamin D and VDBP serum levels in children with CHB and the association of vitamin D and VDBP with HBV replication and hepatic fibrosis.METHODS We enrolled 204 children with CHB admitted to Hunan Children’Hospital in summer and autumn between 2018 and 2019 and 170 healthy controls.CHB patients included:164 hepatitis B e antigen(HBeAg)positive and 40 HBeAg negative;193 hepatitis B surface antigen(HBsAg)positive and 11 HBsAg negative;164 with detectable HBV deoxyribonucleic acid(DNA)and 40 with undetectable HBV DNA;131 with HBV genotype B and 23 with HBV genotype C;and 27 without hepatic fibrosis and 97 with hepatic fibrosis.Serum levels of 25(OH)D,VDBP,liver function markers,and other clinical parameters were collected to analyze their association with vitamin D and VDBP.Mann-Whitney U test,Kruskal-Wallis H test,or t test was used to analyze serum 25(OH)D and VDBP levels in different groups.Spearman rank correlation test was utilized to analyze the correlation of 25(OH)D and VDBP with other markers.Statistically significant factors determined by univariate analysis were further analyzed by binary multivariate logistic regression analysis.P<0.05 was considered statistically significant.RESULTS Children with CHB had lower serum 25(OH)D(56.64±17.89 nmoL/L)and VDBP[122.40(70.74-262.84μg/L)]levels than healthy controls had(P<0.001).Serum 25(OH)D and VDBP levels were significantly different among the different grades of hepatic fibrosis(P<0.05).VDBP levels in children with HBV genotype C,HBsAg,HBeAg,and detectable HBV DNA were significantly lower than those in children with HBV genotype B,no HBsAg,no HBeAg,and undetectable HBV DNA(P<0.05).Serum 25(OH)D level was negatively correlated with age and serum total bilirubin level(r=-0.396 and-0.280,respectively,P<0.001).Serum VDBP level was negatively correlated with HBV DNA(log10 IU/mL)(r=-0.272,P<0.001).Serum 25(OH)D level was not correlated with VDBP level(P>0.05).Univariate(P<0.05)and multivariate logistic regression analysis showed that low level of 25(OH)D(odds ratio=0.951,95%confidence interval:0.918-0.985)and high level of HBV DNA(odds ratio=1.445,95%confidence interval:1.163-1.794)were independently correlated with hepatic fibrosis(P<0.01).CONCLUSION Serum levels of 25(OH)D and VDBP are decreased in children with CHB.Serum VDBP level is negatively correlated with HBV replication.Low level of 25(OH)D is independently associated with hepatic fibrosis in children with CHB.There is no significant association between serum levels of 25(OH)D and VDBP.     

Noninvasive assessment of liver damage in chronic hepatitis B

AIM:To evaluate the efficacy of the aspartate aminotransferase/platelet ratio index(APRI)and neutrophillymphocyte(N/L)ratio to predict liver damage in chronic hepatitis B(CHB).METHODS:We analyzed 89 patients diagnosed with CHB by percutaneous liver biopsy and 43 healthy subjects.Liver biopsy materials were stained with hematoxylin-eosin and Masson’s trichrome.Patients’fibrosis scores and histological activity index(HAI)were calculated according to the Ishak scoring system.Fibrosis score was recognized as follows:F0-1 No/early-stage fibrosis,F2-6 significant fibrosis,F0-4 non-cirrhotic and F5-6 cirrhotic.Significant liver fibrosis was defined as an Ishak score of≥2.APRI and N/L ratio calculation was made by blood test results.RESULTS:The hepatitis B and control group showed no difference in N/L ratios while there was a significant difference in terms of APRI scores(P<0.001).Multiple logistic regression analysis revealed that the only independent predictive factor for liver fibrosis in CHB was platelet count.APRI score was significantly higher in cirrhotic patients than in non-cirrhotic patients.However,this significance was not confirmed by multiple logistic regression analysis.The optimum APRI score cut-off point to identify patients with cirrhosis was 1.01with sensitivity,specificity,positive predictive value and negative predictive value of 62%(36%-86%),74%(62%-83%),29%(13%-49%)and 92%(82%-97%),respectively.In addition,correlation analyses revealed that N/L ratio has a negative and significant relationship with HAI(r=-0.218,P=0.041).CONCLUSION:N/L ratio was negatively correlated with HAI.APRI score may be useful to exclude cirrhosis in CHB patients.     

Expression of platelet-derived growth factor-BB in liver tissues of patients with chronic hepatitis B

AIM:To study the relationship between expression of plateletderived growth factor-BB (PDGF-BB) and fibrogenesis in chronic hepatitis B.METHODS:Hepatic tissues from 43 patients with chronic hepatitis B were embedded in paraffin.The sections were stained with HE and picric acid-sirius red to determine inflammatory activity and fibrosis stages. PDGF-BB expression was detected by immunohistochemistry and assessed semiquantatively. Levels of serum hyaluronic acid (HA),pro-collagen Ⅲ(PCⅢ), collagen Ⅳ (Ⅳ-C) and laminin (LN) were examined by radioimmunoassay (RIA).RESULTS:The expression level of PDGF-BB was found to be positively correlated with inflammatory activity, fibrosis stage and grade of histological findings (τ=0.58,0.55,0.55,P&lt;0.01).The positive correlation was also observed between tissue level of PDGF-BB expression and contents of HA, PCⅢ, Ⅳ-C and LN in the circulation (r=0.52,0.32,0.40,0.33, P&lt;0.05).CONCLUSION: PDGF-BB may play some role in the development and progression of liver fibrosis.     

CT perfusion at early stage of hepatic diffuse disease

AIM: To determine the validity of the non-invasive method of CT perfusion (CTP) in rat model of hepatic diffuse disease. METHODS: Twenty-eight Wistar rats were divided into two groups. Liver diffuse lesions were induced by diethyln-itrosamine in 14 rats of test group. Rats in control group were bred with pure water. From the 1st to 12th wk after the test group was intervened, both groups were studied every week with CTP. CTP parameters of liver parenchyma in different periods and pathologic changes in two groups were compared and analyzed. RESULTS: The process of hepatic diffuse lesions in test groups was classified into three stages or periods according to the pathologic alterations, namely hepatitis, hepatic fibrosis, and cirrhosis. During this period, hepatic artery flow (HAF) of control group declined slightly, mean transit time (MTT), blood flow (BF) and volume (BV) increased, but there were no significant differences between different periods. In test group, HAF tended to increase gradually, MTT prolonged obviously, BV and BF decreased at the same time. The results of statistical analysis revealed that the difference in the HAF ratio of test group to control group was significant. The ratio of BV and BF in test group to control group in stage of hepatitis and hepatic cirrhosis, hepatic fibrosis and early stage of hepatic cirrhosis was significantly different, but there was no significant difference between hepatitis and hepatic fibrosis. The main pathological changes in stage of hepatitis were swelling of hepatic cells, while sinusoid capillarization and deposition of collagen aggravated gradually in the extravascular Disse's spaces in stage of fibrosis and early stage of cirrhosis. CONCLUSION: The technique could reflect some early changes of hepatic blood perfusion in rat with liver diffuse disease and is valuable for their early diagnosis.