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Inflammatory activation of hepatic macrophages plays a primary role in drug-induced liver injury (DILI). However, the exact mechanism underlying DILI remains unclear.A total of 328 DILI patients and 80 healthy individuals were prospectively enrolled in this study. The DILI patients were categorized into subgroups based on either disease severity or histopathological patterns. Plasma soluble CD163 (sCD163) and hepatic CD163 were examined to determine hepatic macrophage activation, and CD8, CD20, and MUM-1 were assessed to determine cellular immunity using immunohistochemistry. The lipopolysaccharide (LPS) pathway proteins [e.g. LPS, soluble CD14 (sCD14), and LPS-binding protein (LBP)] were measured using enzyme-linked immunosorbent assay.Plasma sCD163 levels were nine-fold higher in DILI patients than in healthy controls at the baseline, but significantly decreased at the 4-week follow-up visit after treatment. The numbers of hepatic macrophages, B cells, and plasma cells were significantly higher in the liver tissues from DILI patients than those from healthy controls. Furthermore, the baseline levels of LPS pathway proteins in the DILI patients were significantly higher than those in the controls. Notably, these proteins significantly decreased at the 4-week follow-up visit but remained significantly higher than the levels for the controls.Hepatic inflammation in DILI involves the activation of hepatic macrophages and cellular immunity, in which the LPS pathway likely plays a role, at least in part. As such, this study has improved our understanding of the pathological mechanisms for DILI and may facilitate the development of better treatments for patients with DILI.

Primary biliary cholangitis (PBC) is an autoimmune disease characterized by chronic destruction of the bile ducts. A major unanswered question regarding the pathogenesis of PBC is the precise mechanisms of small bile duct injury. Emperipolesis is one of cell‐in‐cell structures that is a potential histological hallmark associated with chronic hepatitis B. This study aimed to clarify the pathogenesis and characteristics of emperipolesis in PBC liver injury. Sixty‐six PBC patients, diagnosed by liver biopsy combined with laboratory test, were divided into early‐stage PBC (stages I and II, n = 39) and late‐stage PBC (stages III and IV, n = 27). Emperipolesis was measured in liver sections stained with haematoxylin‐eosin. The expressions of CK19, CD3, CD4, CD8, CD20, Ki67 and apoptosis of BECs were evaluated by immunohistochemistry or immunofluorescence double labelling. Emperipolesis was observed in 62.1% of patients with PBC, and BECs were predominantly host cells. The number of infiltrating CD3+ and CD8+ T cells correlated with the advancement of emperipolesis (R 2 = 0.318, P

Soluble cluster of differentiation 40 (sCD40) is proteolytically cleaved from membrane-bound CD40 and binds to CD154, thereby inhibiting CD40-CD154-mediated immune responses. The aim of the present study was to clarify the role of sCD40 in chronic hepatitis B (CHB). The sCD40 levels in sera from 132 patients with CHB and 33 healthy individuals were retrospectively measured. sCD40 concentrations in patients with CHB were higher than those in healthy controls, and sCD40 levels correlated positively with serum levels of the liver dysfunction biomarkers alanine transaminase (ALT) and aspartate transaminase (AST). sCD40 concentrations increased with a rise in the severity of liver necroinflammation and fibrosis. Patients with >75% liver tissue staining positive for hepatitis B virus (HBV) antigen expression showed significantly lower sCD40 levels than those who stained negative for the HBV antigen. The area under the receiver operating characteristic curve of sCD40 was greater than that of ALT and AST; thus, sCD40 levels have a high diagnostic accuracy for detecting severe liver inflammation in patients with CHB, and could serve as an immunological marker of hepatic tissue injury.

Objectives: MicroRNA-122 has been shown to be crucial for efficient HCV RNA replication in vitro. Pretreatment intrahepatic microRNA-122 levels in chronic hepatitis C (CHC) patients have been associated with the outcomes of interferon therapy. Here, we determined microRNA-122 serum levels in CHC patients and healthy donors using an absolute quantification approach and evaluated the correlation with liver inflammation grades and serum alanine aminotransferase (ALT) levels. Methods: Serum samples were collected from 105 treatment-naive CHC patients, 11 acute hepatitis patients, and 33 healthy donors. Serum microRNA-122 was measured using the TaqMan RT-qPCR. The cycle threshold values were converted to copy numbers by drawing a standard curve using a chemical synthetic standard. For accurate quantification, copy numbers were further normalized according to the recovery ratios of spiked-in cel-miR-39. Results: Serum levels of microRNA-122 were significantly higher in acute hepatitis and CHC patients than in healthy donors (p

Objective To explore the expression of cyclin D1, p21WAF1, p53 and Ki-67 protein in hepatocellular carcinoma (HCC) and its relationship with prognosis of HCC patients. Methods Liver specimens were collected from 80 HCC patients who received hepatectomy in the General Hospital of PLA from Jan. 2000 to Jan. 2005, and the expressions of cyclin D1, p21WAF1, p53 and Ki-67 protein were determined by immunohistochemical staining (EliVision method) to investigate the relationship between the protein expressions and the clinicopathologic characteristics of HCC, and their relationship with the survival rate of the patients was analyzed. Results The positive expression rates of cyclin D1, p21WAF1, p53 and Ki-67 in HCC were 38.8%, 40.5%, 65.4% and 80.0% respectively, and they were significantly higher than those in matched normal tissues (19.0%, 11.5%, 0.0% and 6.3% respectively, P

Background and Aim Liver transplantation (LT) for hepatitis B virus (HBV)-related disease can be complicated by HBV recurrence. The aim of this study was to evaluate the risk factors, prophylaxis treatment, and histological characteristics of HBV recurrence after LT when using long-term, low-dose hepatitis B immunoglobulin (HBIG) plus nucleoside analog (lamivudine [LAM] or entecavir [ETV]). Methods Retrospective data from 253 adult LT patients using long-term, low-dose HBIG plus nucleoside analog after LT, for a mean treatment duration of 1–72 months, were collected from a single center in Beijing, China. Univariate analyses were conducted to determine the association among gender, age, hepatocellular carcinoma, hepatitis B e antigen-positive status, HBV-DNA level and tyrosine-methionine-aspartate-aspartate (YMDD) mutations on HBV recurrence in these patients. Results Overall, the HBV recurrence rate was 6.32% (16/253). There was no significant difference in the survival rate between the HBV recurrence and non-recurrence groups. Risk factors for HBV recurrence were: hepatitis B e antigen positivity, HBV-DNA > 105 copies/mL, hepatocellular carcinoma, and YMDD mutation. Sixteen patients receiving LAM had HBV recurrence (16/169; mean treatment duration: 61.8 ± 18.3 months). No HBV recurrence occurred in patients receiving ETV after LT (0/84; mean treatment duration: 57.1 ± 15.9 months). Differences in rate of mortality and HBV recurrence were not significant between the two groups. Conclusions LT is an effective treatment for HBV-related end-stage liver disease. The combination of ETV and intramuscular HBIG for HBV recurrence prophylaxis after LT was more effective than LAM, especially in Chinese patients with HBV recurrence risk factors.

Objective To explore epidemiological, serological and histopathological (by liver biopsy) features of liver diseases, and clinical manifestations in patients of the Chinese armed forces. Methods The clinical data of 231 cases of military patients with liver diseases in our hospital were retrospectively analyzed in terms of their age, gender, location of enlistment, services, official rank, clinical manifestation, and laboratory examination, and also pathological characteristics of liver biopsy. Results Among the 231 hospitalized military patients, 202 were male and 29 were female. The age at onset of the disease ranged from 18 to 73 years (mean age 29.7±9.1). Higher morbidity (48.1%) was found in the 18-25 year age bracket, while lower (only about 7.4%) in above 55-year-old age bracket. Virus infection accounted for 68.0% and non-virus infection accounted for 32.0%. About 64.9% of the patients suffered from chronic liver disease, while 35.1% from acute liver disease. In addition, the prevalence of liver disease was as high as 47.2% in the soldiers, slightly higher than that in the officers (about 38.1%). Transmission of the disease between comrades in arms accounted approximately for 14.0%. Conclusions The mean age of onset of liver disease in military personnel is younger, ranging from 18 to 25 years old predominantly, and the incidence is gradually decreased along with the age. The prevalence of liver disease may be higher in soldiers than in officers. There is a higher percentage of virus infection-associated liver ailment and chronic liver ailment. For acute liver ailment, pathological diagnosis by liver biopsy should be made, and timely therapeutic measures should be taken to prevent transformation of acute to chronic stage.

Objective To establish a method of detecting HBV covalently closed circular DNA(cccDNA) in micro-formalin fixed paraffin imbedding(FFPE) liver biopsy samples.Methods FFPE liver biopsies from 37 patients with chronic hepatitis B were studied.The intrahepatic HBV DNA was extracted and pre-treated with plasmid-safe ATP-dependent DNAse(PSAD),and then amplified by rolling circular amplification(RCA).The HBV cccDNA was quantitatively detected by Taqman real-time PCR with primers located on both sides of the gap of HBV DNA.The human β-actin gene served as the internal control.The sensitivity was tested by serially diluting the DNA templates with known concentrations.The repeatability and stability were evaluated with inter-assay and intra-assay.The level of intrahepatic HBV cccDNA,HBV total DNA,serum HBV DNA and ALT were also compared to find the relations between them.Results The quantitative detection method of cccDNA in micro-FFPE liver samples was successfully set up with considerable sensitivity,stability and specificity.The intrahepatic cccDNA level was significantly higher in the HBeAg-positive patients than in the HBeAg-negative patients(P < 0.05).The intrahepatic HBV cccDNA level was positively correlated with the serum and intra-hepatic HBV DNA level(r=0.539,P=0.001).Conclusion The assay established by present study is fit for the detection of HBV cccDNA in micro-FFPE liver biopsies.

Objective To investigate the relationship of the proliferation and activation of immunocytes in gastric mucosa with the disease progression in HIV-infected individuals.Methods Thirty-six HIV-infected individuals,and 10 persons without HIV infection serving as control,were enrolled in present study,and biopsy of gastric mucosal tissues was performed by fiberoptic gastroscopy.Immunohistochemistry(SP) was used to detect the expressions of CD38 and Ki-67 in gastric mucosa.Masson stain was used to observe collagen deposition in gastric mucosa.Results Compared with the non-infected controls,the expression of CD38 remarkably increased in HIV-infected individuals(97.384±11.573 vs 86.215±12.396,P < 0.01),but there was no difference between asymptomatic HIV carriers and AIDS patients(82.954±10.725,79.982±12.443,P > 0.05).In HIV-infected individuals,Ki-67 positive cells were mainly located in gastric foveola,lamina propria gland and stroma.As compared with the non-infected controls,Ki-67 positive immunocytes were remarkably decreased in stroma(20.933±2.719 vs 29.091±21.949,P < 0.01),but there was also no significant difference between asymptomatic HIV carriers and AIDS patients(27.985±4.650,29.328±7.343,P > 0.05).The Ki-67 positive cells were mostly lymphocytes in the lymphoid follicle of gastric mucosa in asymptomatic HIV carriers,while connective tissue cells such as fibroblasts were predominant in AIDS patients.There was more abundant collagen deposition in stroma and lymphatic follicles in AIDS patients than in asymptomatic HIV carriers.In AIDS patients,the follicle fibrous septum was found to be increased and widened in some region.Conclusion After HIV infection,the disequilibrium of activation and proliferation of immunocytes,as well as collagen deposition in mucosa,are probably the main pathogenetic factors of AIDS.

Objective To investigate the clinical and pathological characteristics of hemochromatosis(HC),and provide references for HC diagnosis and treatment.Methods Liver specimens were obtained via needle biopsy from 20 cases of HC.Histological specimens were stained with haematoxylin eosin.Pathological changes of liver tissues were analyzed together with the clinical data.Results Ten cases of hereditary hemochromatosis(HHC) and 10 cases of secondary hemochromatosis(SHC) were randomly selected.Fatigue(18/20),hepatomegalia(18/20) and splenomegalia(17/20)were the common clinical manifestations.The 20 HC cases characterized by iron overload and fibrosis may be divided into HHC type(17 cases) and non-HHC type(3 cases) according to the region of iron deposition.All the 10 cases of HHC showed HHC type,while 7 of the 10 SHC cases showed HHC type,and the other 3 SHC cases showed non-HHC type.Steatosis,eosinophile granulocyte infiltration and vacuolus nucleus were also observed frequently in the liver tissues of HC,and their distribution coincided with the region of iron deposition.Statistically,fibrosis was significantly associated with iron deposition and serum iron in HHC patients(P < 0.05),but not associated with steatosis and duration of HHC.Additionally,fibrosis was not associated with iron deposition,serum iron,steatosis and duration of SHC in SHC patients.Conclusions The final diagnosis of HC depends mainly on histological changes in liver tissues.Meanwhile,it is necessary to distinguish HHC from SHC according to case history and biochemical detection.HHC might be a metabolic disease with multi-organ damage due to the disruption of homeostasis by iron overload.To avoid multi-organ failure,patients with HHC should be diagnosed and treated as early as possible.

Aims: This study attempts to characterize the feature of immunologically competent cells (ICCs) and evaluate its clinical implication in patients with acute-on-chronic liver failure (ACLF) in relation to chronic hepatitis B virus (HBV) infection. Methods: Circulating ICCs were examined in ACLF patients (n = 75), as well as in patients with hepatitis B (CHB, n = 31), CHB-related liver cirrhosis (LC, n = 36), and normal controls (NC, n = 30). Intrahepatic ICCs in some patients were further analyzed via immunohistochemical and flow cytometric assays. Results: Total lymphocytes, CD4+ T cells, CD8+ T cells, and NK cells in circulation were numerically lower in the ACLF and LC groups compared to the CHB and NC groups. Importantly, the number of these cells was significantly lower in non-surviving ACLF patients compared with surviving ACLF patients. In comparison to NC, ACLF patients displayed a significantly higher ratio of liver-infiltrating CD4+ T-cell frequency than its circulating counterpart, suggesting that the possiblility of the ICCs compartmentalization from the peripheral blood into the liver in ACLF. Immunohistochemical analysis showed that intrahepatic CD4+ cells, CD8+ cells, and CD56+ cells were significantly higher in the ACLF group compared with the other three groups, suggesting a stronger cellular immune response-mediated inflammation in ACLF group than other patient groups. Conclusions: The abnormal prevalence of circulating and intrahepatic ICCs possibly acts as an important factor that may drive the progression of HBV-related ACLF.

Goals This study attempts to determine expressions of intrahepatic proinflammatory and anti-inflammatory cytokines and their secreting immunocytes to evaluate their roles in the pathogenesis of acute-on-chronic liver failure (ACLF) in chronically hepatitis B virus (HBV)-infected patients. Background ACLF generally affects patients with established, compensated chronic liver diseases who develop an acute deterioration in liver function. In China, HBV-associated ACLF patients account for more than 80% of ACLF patients owing to a high prevalence of chronic HBV infection. Clinical observation showed that the deterioration of this disease may correlate with host immune responses, but related underlying mechanism remains largely unknown. Study In situ expressions of interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), interleukin-10 (IL-10), and their secreting CD4, CD8 T cells, and Kupffer cells (KCs) were analyzed in the livers of patients with ACLF, chronic hepatitis B (CHB), and normal controls (NC) using immunohistochemistry. Results Intrahepatic proinflammatory IFN-gamma and TNF-alpha expressions were markedly up-regulated in ACLF compared with CHB and NC. However, similar anti-inflammatory IL-10 expressions were observed in ACLF and CHB. IFN-gamma overexpression correlated significantly with increased CD4 and CD8 T-cell accumulation. TNF-alpha up-regulation also correlated significantly with increased KCs. Conclusions The imbalanced expression of proinflammatory and anti-inflammatory cytokines and increased accumulation of CD4, CD8 T cells, and KCs may contribute to immunopathogenesis in HBV-infected ACLF.

The ongoing Ebola outbreak poses an alarming risk to the countries of West Africa and beyond. On August 8, 2014, the World Health Organization (WHO) declared the cross-country Ebola outbreak a Public Emergency of International Concern. China has had no confirmed cases of Ebola. In this paper, virologic characteristics, pathogenesis, clinical manifestations, laboratory examination and prophylactic vaccines and therapeutic drugs of Ebola are summarized. Importantly, active responses and actions from China are introduced. Moreover, the key issues in the future prevention and control of Ebola were also addressed.

Objective To explore the clinical and pathological features of congenital hepatic fibrosis with Caroli’s disease.Methods The clinical characteristics of 6 patients with congenital hepatic fibrosis and Caroli’s disease were analyzed,and the histological features of hepatic tissue were observed by using hematoxylin and eosin staining and histochemical staining.Results The mean age of the 6 patients was 7.8 years,and the sex ratio(male to female) was 2:4.The main clinical manifestations were hematemesis(5/6),hypodynamia(5/6) and hepatosplenomegaly(5/6).Three of the 6 patients were complicated with portal hypertension,2 cases complicated with diffuse parenchyma lesion in the double kidneys,and 1 case showed abnormal hepatic function.The main histological features were fibrosis of periportal area,epithelial tissue proliferation of interfibrous bile ducts,capsular ectasia of intrahepatic bile ducts,cholestasis and inflammatory infiltration.Pseudolobule formation in hepatic tissue may observed,but most of the hepatic plates arranged still in order.Conclusions Congenital hepatic fibrosis with Caroli’s disease is presented in adolescent.The diagnosis of the disease is mainly based on periportal fibrosis and capsular ectasia of intrahepatic bile ducts combined with clinical data.

Objective To investigate the clinical and pathological characteristics of vanishing bile duct syndrome(VBDS),and provide references for clinical diagnosis.Methods Twenty four children with VBDS diagnosed by liver biopsy were collected in 302 Hospital of PLA from 2007 to 2009.Liver tissues were stained with haematoxylin eosin staining.The clinical and pathological characteristics were analyzed.Results The mean age of all patients(mean SD) was 4.08±3.11 years and the sex ratio(male to female) was 2.4:1.The most common symptoms were jaundice and pruritus in 24 children with VBDS.The mean levels of ALT,AST,ALP,GGT and TBil were 203.45 239.42U/L,238.54 224.11U/L,524.04 300.96U/L,242.17 220.86U/L and 242.17 220.86 μmol/L,respectively.Nineteen of 24 cases were positive for anti-CMV IgM,anti-CMV IgG or CMV PP65.Five cases were undergone the operation of biliary atresia.The main pathological characteristic of VBDS was atresia or disappearance of interlobular bile ducts to various degrees.Moreover,histological changes varied with respect to different causes.Obvious inflammatory infiltration in the portal tracts and hepatocytes damage were more common in VBDS caused by cytomegalovirus(CMV) infection.Conclusions The incidence of VBDS is higher in boys than in girls.CMV infection plays an important role in the progress of VBDS.Different pathological characteristics may exist in VBDS caused by different etiological factors.

Objective To explore the histological morphous,immunohistochemical and clinical features,differential diagnosis and prognosis of hepatic inflammatory angiomyolipoma(AML).Methods The clinical and pathological data of 5 patients with inflammatory hepatic AML were retrospectively analyzed followed by follow-up.The typical tissue was observed by immunohistochemical staining.Results Five patients(1 male and 4 females) aged from 21 to 48 years.Tumors were situated in left lobe of liver in 3 patients,and in right lobe in 2 patients.The tumors had a diameter of 5.5-10.0cm averaged 7.5cm.Generally,the tumors were grey-white on cut surface,without hemorrhage,necrosis and cystic degeneration.Microscopically,the tumors were consisted of spindle and epithelioid myocytes,irregular thick-walled blood vessels and adipose tissue in different proportion.A prominent infiltration of inflammatory cells,including lymphocytes,plasmocytes and histiocytes,in the background was noted in all cases.No obvious atypia and mitotic figure,and no hemorrhage,necrosis and cystic degeneration were noted in the tumor cells.It was immunohistochemically revealed that the tumor cells of all the 5 patients were positive in HMB-45,Melan-A,SMA and Vimentin,while negative in CD21,CD35,CK18,CK19,hepatocyte paraffin-1,AFP,EMA and CK.All the patients were undergone local hepatectomy without recurrence during a 3 to 9 years follow-up.Conclusion Inflammatory hepatic AML is one of the sub-types of AML,demonstrating a similar benign biologic behavior with classical AML.

Background This study aimed to investigate the relationships of intrahepatic cccDNA with serum HBsAg and with HBV DNA in treatment-naive patients throughout acute and chronic HBV infection. Methods A total of 120 patients who had a liver biopsy were enrolled, including 19 with acute hepatitis B (AHB), and 101 patients with chronic HBV infection (CHB) of whom were 10 in immune-tolerant (IT) phase, 59 in immune-clearance (IC) phase, 8 in low-replicative (LR) phase, and 24 in HBeAg-negative hepatitis (ENH) phase. Intrahepatic cccDNA, serum HBsAg and serum HBV DNA levels were comparatively analyzed. Results The median intrahepatic cccDNA levels were 0.18 4.80, 3.81, 0.22 and 0.97 copies/cell for patients with AHB, CHB-IT, CHB-IC, CHB-LR, and CHB-ENH, respectively. In AHB patients, intrahepatic cccDNA was positively correlated with serum HBsAg (r = 0.665, P = 0.003), as well as serum HBV DNA (r = 0.536, P = 0.022). In CHB patients, intrahepatic cccDNA was positively correlated with serum HBsAg in the IC phase (r = 0.392, P = 0.005), and with serum HBV DNA in the IC phase (r = 0.301, P = 0.036) and ENH phase (r = 0.588, P = 0.013). HBV replicative efficiency, defined as the ratio of serum HBV DNA to intrahepatic cccDNA, was obviously lower in AHB and CHB-LR patients than in CHB-IT, CHB-IC and CHB-ENH patients (0.70 and 0.53 vs. 1.12, 1.09 and 0.99, P

Liver transplantation (LT) for hepatitis B virus (HBV)-related disease can be complicated by HBV recurrence. The aim of this study was to evaluate the risk factors, prophylaxis treatment, and histological characteristics of HBV recurrence after LT when using long-term, low-dose hepatitis B immunoglobulin (HBIG) plus nucleoside analog (lamivudine [LAM] or entecavir [ETV]).Retrospective data from 253 adult LT patients using long-term, low-dose HBIG plus nucleoside analog after LT, for a mean treatment duration of 1-72 months, were collected from a single center in Beijing, China. Univariate analyses were conducted to determine the association among gender, age, hepatocellular carcinoma, hepatitis B e antigen-positive status, HBV-DNA level and tyrosine-methionine-aspartate-aspartate (YMDD) mutations on HBV recurrence in these patients.Overall, the HBV recurrence rate was 6.32% (16/253). There was no significant difference in the survival rate between the HBV recurrence and non-recurrence groups. Risk factors for HBV recurrence were: hepatitis B e antigen positivity, HBV-DNA 10(5) copies/mL, hepatocellular carcinoma, and YMDD mutation. Sixteen patients receiving LAM had HBV recurrence (16/169; mean treatment duration: 61.8 ± 18.3 months). No HBV recurrence occurred in patients receiving ETV after LT (0/84; mean treatment duration: 57.1 ± 15.9 months). Differences in rate of mortality and HBV recurrence were not significant between the two groups.LT is an effective treatment for HBV-related end-stage liver disease. The combination of ETV and intramuscular HBIG for HBV recurrence prophylaxis after LT was more effective than LAM, especially in Chinese patients with HBV recurrence risk factors.

To explore whether the cellular apoptosis susceptibility (CAS) protein could serve as a pathologic marker for HCC diagnosis and the roles of CAS expression in HBV infection associated HCC.The expression of CAS protein in HCC and its paracarcinoma tissues, non-tumor liver cirrhosis and hepatitis tissues were detected by immunohistochemistry. Meanwhile, HBsAg, HBcAg and HBV DNA in HCC tissues with HBV infection were examined by immunohistochemistry and in situ hybridization respectively.The expression of CAS protein was significantly higher in HCC than in its paracarcinomas tissues (P0.01), and higher in paracarcinomas tissues than in non-tumor liver cirrhosis and hepatitis tissues (P0.01). Poorly differentiated tumors immunochemically stained stronger than moderately or well differentiated (P0.01). CAS protein expression was significantly higher in HBV-infected HCC tissues than that of in non-HBV infection (P0.01). Meanwhile, in HBV-infected HCC tissues, the staining intensity score of CAS protein in HBV DNA positive HCC tissues was significantly higher than HBV DNA negative tissues (P0.05).Higher expression of CAS protein is found in HCC tissues,and the intensity of CAS protein expression is related closely to tumor differentiation. We suggested that CAS protein might serve as a marker for HCC diagnosis and differentiation estimation, and deduced that CAS might play an important role in the initiation of HBV infection associated HCC through upregulating expression of CAS.

To quantitatively detect intrahepatic HBV covalently closed circular DNA (cccDNA) and serum HBsAg; and to analyze the relationship between the two parameters and with serum HBV DNA level.Intrahepatic cccDNA (copies/cell) was quantitated by plasmid-safe ATP-dependent Danes (PSAD) digestion in combination with rolling circle amplification and gap-spanning selective real-time PCR assay using formalin fixed paraffin-embedded liver biopsy samples. HBsAg was measured by chemiluminescence's reagent manufactured by Abbott Company using sera sampled at time-point of liver biopsy.Intrahepatic cccDNA level was positively correlated with serum HBsAg level (r = 0.459, P0.001), but not correlated with serum HBV DNA level. Serum HBsAg level was positively correlated with serum HBV DNA level (r = 0.328, P = 0.015), and reversely correlated with HBV replicative efficiency defined as the ratio of serum HBV DNA to cccDNA (r = -0.373, P = 0.005).In patients with chronic hepatitis B, intrahepatic cccDNA level is correlated with serum HBsAg level. The two parameters combined with serum HBV DNA may comprehensively reflect HBV replication activity and help evaluation of antiviral therapeutic efficacy.

In the title compound, C21H25NO4, the dihydropyridine ring adopts a flattened boat conformation. The N atom and the sp3 C atom deviate in the same direction from the mean plane of the other four C atoms, by 0.269 (6) and 0.111 (6) Å, respectively. This mean plane is inclined to the 4-methoxyphenyl ring by 87.3 (5)°. The cyclohexenone ring has a sofa conformation with the C atom bearing the methyl groups deviating from the mean plane through the other five C atoms by 0.628 (6) Å. There is a short C—H...O hydrogen bond in the molecule. In the crystal, molecules are linked by an N—H...O hydrogen bond to form chains propagating along the c-axis direction.

To investigate the frequencies and distribution of CD4+ T cells and CD8+ T cells as well as the changes of immune activation in gastric mucosa of HIV-infected individuals.42 HIV-infected individuals were recruited into this investigation, and 36 patients had definite diagnosis of clinical stage. Biopsy of gastric mucosal tissues was performed by fiberoptic gastroscope including 10 normal people as a control group. Then, immunohistochemistry was used to detect expression of CD4, CD8 and CD38 in gastric mucosa, and the distinctions among three groups were analyzed with LEICA Qwin image analysis system.(1) Compared with asymptomatic HIV carriers and control group, CD4 T cells remarkably decreased in the gastric mucosa of AIDS patients (P0.01). In gastric mucosa of asymptomatic HIV carriers, there were still some CD4+ T cells in lymphoid follicles and stroma where CD4+ T cells were unevenly distributed, the frequency of CD4+ T cells was not significantly different between asymptomatic HIV carriers and control group (P0.05); (2) Phenomenon of CD8+ T cells infiltrating mucosal epithelium and gland was general in HIV-infected individuals. CD8+ T cells took on local excessive hyperplasia in gastric mucosa of some individuals. As compared with control group, CD8+ T cells markedly increased in gastric mucosa of infected individuals (P0.01), but the distinction of asymptomatic HIV carriers and AIDS patients was not significant (P0.05); (3) CD38-expressing cells mainly distributed over gastric mucosal surface to superficial layer(1/3-2/3 layer) of HIV-infected individuals, and was more intensive than control group (Por = 0.01), but there was not noticeable difference between asymptomatic HIV carriers and AIDS patients (P0.05).The frequencies and distribution of gastric mucosal CD4+ T cells of HIV-infected individuals were closely correlated with progression of disease. Disfunction of mucosal immune system which was resulted from HIV infection and injury of CD4+ T cells could be an important cause of CD8+ T cells increasing and CD38-expressing enhancement.

To understand the level and clinical significance of soluble CD40 in patients with chronic hepatitis B.Detecting the concentration of sCD40 from 176 cases with chronic hepatitis B by ELISA and analyzing its relationship with different grades of inflammation and necrosis in liver tissue.sCD40 from patients with chronic hepatitis B was significantly higher than those from healthy. And that the concentration of sCD40 was positive correlation with severe clinical disease and liver inflammation and necrosis. In patients whose ALT lower than 80 IU/L and sCD40 higher than 80 pg/ml, it showed that 65.85% cases have high grade of liver inflammation and necrosis, which was significantly higher than patients with sCD40 lower than 80 pg/ ml.The concentration of sCD40 is positively related with the grade of liver inflammation and necrosis. This information could help us to evaluate the status of chronic hepatitis B as an immunological index.

To establish a new grading system to evaluate liver inflammation and necrosis in patients with chronic hepatitis B through clinical biochemical assays.Clinical and pathological data were collected from 255 cases with chronic hepatitis B. 19 biochemical items were analyzed and 5 items were selected for our grading system. Each of the five items was scored 0 to 4 based on the different values. The extent of liver inflammation and necrosis was evaluated according to the total score.ALT, AST, ChE, GGT and TBA were selected for our grading system. The grade of liver inflammation and necrosis was considered less than 2.0 if total score lower than 6, and higher grade was considered with higher total score. The estimated results shared an identity of 82.8% with the real grades of liver inflammation and necrosis. When this grading system was applied to patients with liver inflammation and necrosis equal to or higher than grade 2.0, it exhibited a sensitivity of 83.8%, a specificity of 81.2%, a positive prediction value of 88.6%, a negative prediction value of 74.2% in all cases, and 88.0%, 84.7%, 90.6%, 82.4% respectively in patients older than 12 years.Our data suggest that the grading system can be used to evaluate the extent of liver inflammation and necrosis in patients with chronic hepatitis B.

Hepatitis C virus is a hepatotrophic flavivirus infecting 170 million people worldwide and 3.2% people in China. We aim to investigate the clinical outcomes and hepatic histology of patients with hepatitis C in the north of China.Nine patients with hepatitis C, 4 males and 5 females, aged 44 +/- 7 years old, who were infected during transfusion or blood donation underwent follow-up for 13-27 years. We performed liver histological examination by biopsy, ultrasonography, and biochemical examination of alanine transaminase (ALT), aspartate transaminase, and total bilirubin. The serum anti-HCV was detected by ELISA, and HCV RNA quantification by PCR.ALT were higher than the normal value at every time point and HCV RNA remained positive (3.57 x 10(5) - 7.21 x 10(6) copies/mL) in all patients. Ultrasonography showed mild and moderated hepatitis (3 and 6 cases respectively). The modified histological activity indices (HAI) were 5.0-8.5 and fibrosis scores were 1-4. The ALT value was higher in the moderate cases than in the mild cases, and higher in those with higher HAI. The viral load was higher in the patients infected by the virus of the genotype 1b than in the patients infected by the virus of genotype 2.The clinical outcomes and hepatic histology of patients with hepatitis C in north of China seem not very severe.

To study the clinical and pathological features of drug-induced liver injury (DILI).Liver specimens were obtained through needle biopsies from 100 patients with DILI. The histological preparations of the specimens were stained with haematoxylin eosin, several histochemistry methods, and immunohistochemistry stains. The pathological changes of the livers were analyzed together with the patients's clinical data. The patients were divided into two groups, an acute DILI group (n=39) and a chronic DILI group (n=61), based on their clinical courses and histological changes in their livers. In the chronic DILI group, the clinical courses were longer than 6 months and/or fibrosis or cirrhosis occurred in their liver tissues.Among our cases the leading cause of DILI was Chinese herb medicine, accounting for 21% of the 100 cases; steroids induced cases were 11% of the total. 78% of the patients presented elevated serum transaminases and/or jaundice. The degree of transaminases elevation and the frequency of jaundice happening in the acute group were significantly higher than those in the chronic group (P less than 0.05). The histopathological liver changes in these DILI cases included: (1) necrosis commonly occurred in acinar zone 3, (2) abundant neutrophil and/or eosinophil infiltrations, (3) hepatocytic and/or canalicular cholestasis with little or no inflammation, (4) microvesicular steatosis mixed with macrovesicular steatosis, and (5) presentation of epitheloid cell granuloma. There were no significant differences in liver histopathology between the acute and the chronic DILI groups, except that the fibrosis and the ductular proliferation were different.DILI has become a notable liver disease in mainland China, and the use of Chinese herbal medicine must be improved, standardized and regulated more closely.

CD4+CD25+ regulatory T cells (Treg) have been shown to maintain immune tolerance against self and foreign Ags, but their role in persistent viral infection has not been well-defined. In this study, we investigated whether and where CD4+CD25+ Treg contribute to the development of chronic hepatitis B (CHB). One hundred twenty-one patients were enrolled, including 16 patients with acute hepatitis B, 76 with CHB, and 29 with chronic severe hepatitis B. We demonstrated that in chronic severe hepatitis B patients, the frequencies of CD4+CD25+ Treg in both PBMC and liver-infiltrating lymphocytes were significantly increased and there was a dramatic increase of FoxP3+-cell and inflammatory cell infiltration in the liver compared with healthy controls. In CHB patients, circulating CD4+CD25+ Treg frequency significantly correlates with serum viral load. In acute hepatitis B patients, circulating CD4+CD25+ Treg frequency was initially low and with time, the profile reversed to exhibit an increased number of circulating Treg in the convalescent phase and restored to normal levels upon resolution. In PBMC taken from infected patients, depletion of CD4+CD25+ Treg led to an increase of IFN-γ production by HBV-Ag-stimulated PBMC. In addition, CD4+CD25+ Treg were capable of suppressing proliferation of autologous PBMC mediated by HBV Ags, which probably reflects the generation of HBV-Ag-specific Treg in circulation and in the liver of HBV-infected patients. Together, our findings suggest that CD4+CD25+ Treg play an active role not only in modulating effectors of immune response to HBV infection, but also in influencing the disease prognosis in patients with hepatitis B.

To investigate the relation of the viral markers in serum and those expressed by hepatocytes to pathological lesions of hepatic tissue in patients with chronic hepatitis B.The relation of viral markers including HBsAg, HBsAb, HBeAg, HBeAb, HBcAb and HBV DNA in serum of 647 patients with chronic hepatitis B and HBsAg, HBcAg expressed by hepatocytes in 418 of these patients to pathological lesions of hepatic tissue was determined.Viral markers in serum and those expressed by hepatocytes in patients with chronic hepatitis B were closely correlated with pathological lesions of hepatic tissue.The degree of inflammation and fibrosis in hepatic tissue is milder in serum HBsAg, HBeAb, HBcAb positive and HBV DNA negative patients but more serious in those with negative hepatocytic expression of HBsAg and HBcAg. HBV DNA is not significantly associated with pathological lesions of hepatic tissue.

To investigate the clinical outcomes and hepatic histology of patients with hepatitis C.Nine patients with hepatitis, 4 males and 5 females, aged 44 +/- 7, who were infected during transfusion or blood donation underwent follow-up for 13 approximately 14 years: liver histological examination by biopsy, ultrasonography, biochemical examination of alanine transaminase (ALT), apartate aminotransferase (AST), and total bilirubin (TBIL), serum anti-HCV by ELISA, and HCV RNA quantification by PCR.ALT were higher than the normal value at every time point and HCV RNA remained positive (3.57 x 10(8) approximately 7.21 x 10(9) copies/L) in all patients. Ultrasonography showed mild and moderate hepatitis (3 and 6 cases respectively). The modified histological activity indices (HAI) were 5.0 approximately 8.5 and the fibrosis scores were 1 approximately 4. The ALT value was higher in the moderate cases than in the mild cases, and higher in those with higher HAI. The viral load was higher in the patients infected by the virus of the genotype 1b than in the patients infected by the virus of the genotype 2.The clinical outcomes and hepatic histology of patients with hepatitis C seem not very severe.

To investigate the expression dynamics and significance of matrix metalloproteinase-2 (MMP-2) membrane type-matrix metalloproteinase-2 (MT-MMP-2) in hepatic fibrosis and its reversal counterpart.An experimental CCl4 induced hepatic fibrosis rat model was established by intraperitoneal administration of carbon tetrachloride for 2, 4, 6, 8, 10 weeks, and normal rats were used as a control group. The immunohistochemical methods and in situ hybridization were used to detect MMP-2,MT-MMP-2 mRNA and related antigens in the liver.MMP-2,MT-MMP-2 mRNA and related antigens were expressed in mesenchymal cells and parts of hepatocytes besides active pathological changes, especially in the fibrous septum and portal area. Expression of MMP-2,MT-MMP-2 mRNA and related antigens were increased in hepatic fibrosis and decreased gradually in its reversal counterpart.This study suggested that mesenchymal cells are the main cellular origins of MMPs. The levels of MMP-2 and MT-MMP-2 antigens and gene expression were closely related to hepatic fibrosis. MMP-2 and MT-MMP-2 may play important roles in hepatic fibrosis and its reversal counterpart.

To explore the cut-off period of subclassification and pathological features of severe hepatitis (SH).Based on combined clinical and pathological analyses, the complete clinical and biopsy or autopsy liver tissues data from 196 cases of patients with severe hepatitis were investigated. Meanwhile, proliferative hepatocytes, cholangioepithelia and collagens were identified by a panel of monoclonal antibodies such as those against albumin, cytokeratin 18,19 and collagen I, III with immunohistochemical method.The clinical and pathological analyses indicated the cut-off periods of acute, subacute and chronic SH (ASH,SSH and CSH) were (13.4+/-7.2) d, (77.4+/-69.3) d and (80.5+/-63.2) d, respectively. Among all SH cases, one case of ASH patient presented clinical manifestation and pathological changes of ASH for 21 days, however, one patient with SSH was demonstrated 12 day course by histological examination. The time of cut-off period between ASH and SSH in child cases was shorter than that in adult cases. Histologically, ASH liver tissues showed massive and/or submassive necrosis caused by one attack, with congestive sinusoid frameworks and proliferative cholangioepithelium-like hepatocytes, while SSH liver tissues presented combined fresh and old submassive or massive necrosis caused by multiple attacks, accompanied by obviously proliferative bile ducts and sinusoid framework collapse.However, the pathological changes of CSH showed ASH- or SSH-like lesions on the background of chronic liver injury.Our data indicated that the cut-off period between ASH and SSH is in accordance with the Scheme of Viral Hepatitis Prevention and Therapy, China, published in 2000, but excluded a part of child SH cases. In our study, the authors found a few pathological features in ASH and SSH.

To explore the pathological features and pathogenesis of severe acute respiratory syndrome (SARS) to provide evidence for the clinical treatment and prevention of SARS.Pathological features of 2 cases of full autopsy and 4 cases of needle biopsy tissue samples from the patients who died from SARS were studied by light and electron microscopy. The distribution and quantity of lymphocyte subpopulations in the lungs and immune organs from SARS patients were analyzed by immunohistochemistry. The location and semi-quantitative analysis of SARS coronavirus in the tissue specimens were studied by electron microscopy, in situ hybridization and immunohistochemistry.In total of 6 cases, diffuse alveolar damage and alveolar cell proliferation were common. The major pathological changes of 2 autopsy cases of SARS in lung tissues were acute pulmonary interstitial and alveolar exudative inflammation, and 2 autopsy and one biopsy lung tissues showed alveolar hyaline membrane formation. Terminal bronchiolar and alveolar desquamation of lung tissues in one autopsy and 2 biopsy cases were noted. Among 6 cases, 2 biopsy cases presented early pulmonary fibrosis and alveolar organization. Meanwhile, the immune organs, including lymph nodes and spleens from 2 autopsy cases of SARS whose disease courses were less than 12 days showed extensive hemorrhagic necrosis, reactive macrophage/histocyte proliferation, with relative depression of mononuclear and granulocytic clones in the bone marrows. However, spleen and bone marrow biopsy tissue samples from 4 dead SARS cases whose clinical course lasted from 21 to 40 days presented repairing changes. SARS coronaviruses were mainly identified in type I and II alveolar epithelia, macrophages, and endothelia; meanwhile, some renal tubular epithelial cells, cardiomyocytes, mucosal and crypt epithelial cells of gastrointestinal tracts, parenchymal cells in adrenal glands, lymphocytes, testicular epithelial cells and Leydig's cells were also detected by electron microscopy combined with in situ hybridization. The semi-quantitative analysis of lymphocyte subpopulations revealed that the proportion of CD8+ T lymphocytes were about 80% of the total infiltrative inflammatory cells in the pulmonary interstitium, with a few CD4+ lymphocytes CD3+, CD4+, CD8+ or CD20+ lymphocyte subpopulations were obviously decreased and there was imbalance in number and proportion, while CD57+, CD68+, S-100+ and HLA-DR+ cells were relatively increased in lymph nodes and spleens.Histologically, the pulmonary changes could be divided into acute inflammatory exudative, terminal bronchiolar and alveolar desquamative and proliferative repair stages or types during the pathological process of SARS. SARS coronavirus was found in multi-target cells in vivo, which means that SARS coronavirus might cause multi-organ damages which were predominant in lungs. There were varying degrees of decrease and imbalance in number and proportion of lymphocyte subpopulations in the immune organs of the patients with SARS. However, these changes may be reversible. It was found that cellular immune responses were predominant in the lungs of SARS cases, which might play an important role in getting rid of coronaviruses in infected cells and inducing immune mediated injury.

Liver biopsy plays an important role in accurate diagnosis of various liver diseases in children and liver damages caused by systemic illnesses. This study was designed to evaluate the value of liver biopsy in diagnosis of liver diseases in children and explore the relationship between their pathological changes and clinical manifestations.One-second liver biopsy was performed in 1023 pediatric patients with liver diseases at our department from 1983 to 2000. Diagnosis of viral hepatitis was based on the diagnostic criteria formulated by the Chinese Society of Infectious and Parasitic Diseases in 1995. Inflammatory changes of the liver were graded from 0 to 4 (G0-4).Liver biopsy was performed successfully in 1020 patients including 135 infants and young children, of whom 90% were hospitalized patients with chronic liver diseases. Hepatitis virus was the leading cause for chronic liver diseases, among which hepatitis B was detected in 75.4% of the patients. Sixty-nine patients showed liver impairment induced by disorders relevant to that metabolism, Wilson's disease, and glycogen storage disease. Liver inflammatory injury (G2) was found in 76.4% of the patients with hepatitis B in contrast to 61.2% of adult patients. It was aggravated with age in the patients with hepatitis B and C and peaked at their schooling age. Moderate or severe liver injuries were not seen in infants with chronic hepatitis B. Two infants had chronic hepatitis C. Patients with non-viral hepatitis showed specific, non-specific histological changes and liver cirrhosis.Liver inflammatory injuries are more common in children with hepatitis B than in adult patients, and severe inflammatory changes are seen in children with hepatitis B and C at their school age. Liver injuries induced by non-viral factors seem to be increasing, and liver biopsy in children is safe and feasible in the diagnosis of liver diseases.

To explore the clinical and pathological features and the pathogenesis of primary biliary cirrhosis (PBC) in Chinese Mainland.30 PBC patients were divided into the early group (Scheuer stage I and II, 19 patients) and the late group (Scheuer stage III and IV, 11 patients). The data of clinics and serology were analyzed, and the pathological features of the liver tissues were characterized. The changes of dendritic cells (DCs) and hepatic stellate cells (HSCs) were studied by immunohistochemistry.In all the PBC patients, the rate of the male to the female was 1 to 5, and the average age was 40.6 years. The mean levels of TBiL, ALP and GGT in the sera were (95.9+-88.5) micromol/L, (537.2+-339.2) U/L, and (582.0+-351.2) U/L, respectively. 73.3% patients showed AMA positive, and the level of GGT was positively correlated with the AMA level according to the result of statistical analysis (r=0.778, P=0.000). The symptoms of jaundice and hepatomegaly were presented more commonly in the late group than those in the early group (chi2=5.182, P0.05; chi2=13.659, P0.01, respectively). The main changes of morphology of PBC located in portal tracts. The liver tissues in the early stage of PBC showed the damage of bile ducts and obvious proliferation of small bile ducts. The granulomas, the lymphoid follicles and the foamy cells were found in the liver tissues of PBC (2/19 patients, 12/19 patients, and 10/19 patients in the early stage respectively, while 0/11 patients, 4/11 patients, and 3/11 patients in the late stage respectively). There was significant difference between the early stage and the late stage in presence of the lymphoid follicles and the foamy cells (t=4.489, P0.05; t=4.019, P0.05, respectively). The biliary pigmentary particles were mainly accumulated in the liver cells around the portal tracts in 90.0% PBC patients, and the accumulation of copper and iron increased, compared with that in normal specimens. The DCs and HSCs located mainly in the portal tracts, especially around the damaged bile ducts.There are some clinical and pathological characteristics in the patients with PBC. The level of AMA has no direct relationship with the level of transaminase or bilirubin. The proliferated bile ductules may express the antigens which maybe the target of immune attack. As an antigen-presenting cell, DCs may play an important role in the pathogenesis of PBC.

Investigate the features of outbreak epidemic, clinical disease progression of the first SARS cases in Beijing and evaluate the efficacy of therapeutic regimen.Twenty-nine patients (11 men and 18 women, 20 - 74 years old age range) were diagnosed with infectious SARS and admitted in our hospital from the March 5th to April 14th, 2003 in this study. The data of clinical presentation and disease progression of all the patients including index subject as the infectious SARS resource patient, her family infected members and 21 health care workers were abstracted.The first SARA outbreak in Beijing was characterized with the cluster feature of resource patient family members and health care providers. The incubation period ranged from 2 to 14 days. All the patients had a fever (temperature38 degrees C for over 24 hours) and other manifestations as reported before. Serial chest radiographs showed progressive pathologic air-space disease. Twenty patients showed the severe syndrome with various time ranged from 1 day to 14 days. Two patients died of progressive acute respiratory distress disease. The histologic analysis of one death patient showed diffuse alveolar damage in the two lungs. Twenty-six patients receiving the combined therapy including use of corticosteroid, antiviral ribavirin agents after the onset of symptoms and showed they had an acute self-limited disease course. The oldest patient (74 year old, male) received the healthy convalescent plasma infusion (50 ml) from recovered SARS subject and completely recovered within 21 days, having a shorter disease course.SARS is a kind of new self-limited and acute infectious disease. Early diagnosis, early isolation, early antiviral therapy for patients and efficient prevention for health care providers are urgently recommended. In particular, a combinational therapy of use of antiviral agents, preventive antibacterial antibiotics and pulsed dosage of corticosteroid can efficiently raise the clinical recovery rate and decrease mortality of SARS patients.

To study the clinical, laboratory, and radiologic features of 34 cases of severe acute respiratory syndrome (SARS) in Beijing.All patients were admitted to the isolation wards. Their demographic, clinical, laboratory, and radiologic characteristics were analyzed. Univariate and multivariate analyses were performed.Eight patients came from a family, and 15 patients were medical staff. The mean age of patients was (33.4 +/- 13.4) years. The latent period varied from 2 to 14 days (median 4 days). The most common symptoms were fever (100%), palpitation (91.7%), myalgia (79.2%), headache (70.8%), diarrhea (73.9%) and cough (58.3%). The mean leucocyte count was (4.6 +/- 1.4) x 10(9)/L, and the mean lymphocyte ratio was 0.27 +/- 0.11. 68.4% of the patients had lymphopenia (absolute lymphocyte count1.3 x 10(9)/L). Other common findings included elevated levels of serum alanine aminotransferase, lactate dehydrogenase and erythrocyte sedimentation (76.2%, 28.6% and 47.8%, respectively), and decreased levels of serum iron and albumin (63.2% and 47.8%, respectively). Thirty-two cases had abnormal chest radiographs. In 2 cases in whom typical lung opacities could not be found on the initial plain chest radiographs, thoracic CT proved to be useful. Postmortem examination of 1 patient revealed marked edema with foci of hemorrhage and hyaline membrane formation in the lungs, hemorrhage necrosis and a obvious decline of cells in lymph glands. In a multivariate analysis (Stata 7.0), the independent predictor of an adverse outcome was advanced age (odds ratio per decade of life, 1.6; 95% CI, 1.08 to 2.63; P = 0.007).Fever, lymphopenia, low serum iron and chest radiograph are helpful to diagnose SARS early; age is the independent predictor of an outcome.