Objective To study the disease spectrum,the features of clinical pathology,epidemiology and prognosis of liver diseases in Chinese populations from Jan.1980 to Jun.2008.Methods Twenty-five thousand nine hundred and forty-six patients with liver diseases in a wide spectrum of inhabitants(including 1 448 military patients) from 31 provinces or cities of all over mainland China in recent 28 years were involved in the present study for a retrospective study regarding their clinical,pathological and epidemiological features,including laboratory re-examinations and pathological examination.1 322 patients with liver disease(course lasted from 6 months to 18 years) were followed-up by more than 2 liver biopsies to study the outcome of chronic hepatitis B.Results For all the patients involved,the sex ratio of male to female was 3 to 1,with mean age of 32.3?14.4 years ranging from 41 days to 91 years,and the diseases occurred predominantly between the age of 18 to 37 years.Hebei,Henan,Beijing,Shandong and Shanxi provinces(city) ranked at the fore in the endemic distribution of the diseases.The spectrum of liver disease covered more than 100 kinds of liver diseases,of which 73.05% were infectious liver disease.As a whole,the incidence of both infectious and non-infectious,especially the non-infectious liver diseases became more prevalent since 2000.It was shown that the chronic hepatitis B was the most predominant factor which caused liver failure,liver cirrhosis and liver cancer,and chronic hepatitis C was the second factor.Chronic infection pathological changes were found in the liver tissues in 0.26% patients with hepatitis A and 0.51% patients with hepatitis E.For all the 1 322 followed-up patients with chronic hepatitis B,the incidence of liver cirrhosis and hepatocellular carcinoma was respectively 15.36%(203 cases) and 1.06%(14 cases),and the average progressive period for the changes in pathology was 46.37?16.93 months and 60.29?39.15 months,respectively.Meanwhile,the degree of liver fibrosis increased more than one stage in 188 patients(14.22%),decreased more than one stage in 441 patients(33.36%),and no change in 476 patients(30.01%).Conclusions The liver disease spectrum during recent 28 years in Chinese populations has been essentially identified by a retrospective analysis of a large number of clinical pathological data.The clinical features of predominant liver diseases have been illustrated,and the outcome and transition time of chronic hepatitis B has been elucidated in present study.

Objective To explore the clinical and pathological features of the autoimmune hepatitis (AIH) and primary biliary cirrhosis (PBC) overlap syndrome (AIH-PBC overlap syndrome). Methods The clinical data and liver biopsy specimens from 17 cases of the AIH-PBC overlap syndrome were analyzed and compared with 117 cases of AIH and 85 cases of PBC. The pathological features were as analyzed by histological observation and immunohistochemical staining for CK19, CD3, CD4, CD8, CD20, CD25, CD57 and CD68 in the biopsy liver tissues. Results Of the cases of autoimmune liver disease in this study, 8.4% were diagnosed as AIH-PBC overlap syndrome based on clinical, biochemical, serological and pathological examinations. Among the overlap syndrome cases, the females were in predominance (male∶female was 1∶8.5), and the median age of the patients presenting the clinical onset signs was 39.3 years, and the serological double-positive autoantibodies of ANA and AMA-M2 occupied 52.9%(9/17). Serum levels of ALT, ALP, ?-Glo, AST, TBIL, IgG and IgM were higher in patients with overlap syndrome than those in patients with AIH or PBC, respectively. The pathological findings in the biopsy liver tissues from the patients with overlap syndrome included moderate to severe interface hepatitis with plasma cell predominated mixed-inflammatory cells infiltration in hepatic and portal tracts, as well as the damage and obvious reactive proliferation of small bile ducts. Meanwhile, the amounts of CD3, CD4, CD8, CD20, CD25, CD57 and CD68 positive mononuclear cells increased in the overlap syndrome liver tissues compared with those in single PBC or AIH cases. Conclusion AIH-PBC overlap syndrome is not frequent (8.4%) in this study, but exist among autoimmune liver diseases in China. This overlap syndrome presented both clinical and pathological features of AIH and PBC, and cellular immune mediated injuries might be pivotal role in pathogenesis.

Objective To investigate the expressions of pivotal cytokines such as TNF-? and TGF-?_1, and leptin in human nonalcoholic lipoid hepatitis (NALH), and to explore their potential roles in the pathogenesis of NALH. Methods Liver tissue obtained from 59 patients pathologically diagnosed as nonalcoholic lipoid hepatitis and 10 healthy adult liver tissues were examined in the present study. The respective expression of TNF-?, TGF-?_1 and Leptin was qualitatively and semi-quantitatively determined at the protein expression levels by using immunohistochemical method and image analysis. Results In NALH liver tissue the expression of TNF-? appeared mainly in inflammatory cells and lining cells of liver sinusoids in zone 3, lipogranulomas and portal tract, where obvious inflammation was found. The expression was parallel to different degrees of inflammation in NALH, significant correlation was found between degree of inflammation and fibrosis and extent of expression (P

Objective To explore the expression of leptin and its receptors in liver tissue of nonalcoholic steatohepatitis(NASH)patient and its significance in pathogenesis.Methods Samples of 59 liver biopsies of NASH patients were divided into three groups G,S and F according to their degree of inflammation and steatosis and stage of fibrosis.The expression of leptin and its receptors including Ob-R,the short form of Ob-R(Ob-Ra)and the long form of Ob-R(Ob-Rb)were semi-quantitatively determined at the protein and/or mRNA expression levels by using immunohistochemistry,in situ hybridization and image analysis.Results The expressions of leptin in NASH liver tissues were found in KC,HSC and MNC,which were mainly situated in zone 3 near steatotic hepatocytes and perisinusoidal or portal fibrosis areas.Leptin mRNA(Ob)positive cells were distributed in hepatocytes in zone 3.The expression of leptin at protein and mRNA levels in the NASH liver tissues was significantly increased in NASH compared with normal liver tissues(P

Objective To explore the pathological features and the causative particles of severe acute respiratory syndrome (SARS) for providing evidences of SARS prevention and clinical treatment. Methods A dead case of SARS of China was studied by light microscopy, electron microscopy, histochemical and immunohistochemical stain. Results The major pathological changes of lung in the SARS case were acute pulmonary interstitial exudative and leakage inflammation, with predominant lymphocyte infiltration. The hyaloid membranes were formed in 20%～30% pulmonary alveoli. The diffuse pulmonary epithelial injury was observed, and virus like inclusions were found in about 30% of total alveolar epithelia by histochemical stain, but chlamydia like inclusions were found occasionally. Meanwhile, the extra pulmonary organs, such as lymph nodes and spleen, showed extensive haemorrhagic necrosis inflammation, accompanied macrophage/histocyte reactive proliferation with erythrocytophage. The double adrenal glands also presented focal haemorrhagic necrosis inflammation. Under the electron microscopy observation, virus like particles with 100 ～150 nm diameter and halo or garland envelopes were found in more than 30 % alveolar epithelial cells, endothelial cells in lung tissues, and also in a part of cardiomyocytes, lymphocytes and macrophages in lymph nodes. The virus like particles were mainly located in cytoplasm and dilated reticular endoplasm. In contrast, chlamydia like particles were commonly visualized in multiple extra lung organs such as liver, but very few in the lung. Immunohistochemistry showed the positive reactions in the lung tissues with the serum IgG and/or IgM from the dead case himself and other SARS convalescent stage cases from Guangdong province of China. Conclusion In the severe SARS case, predominant acute interstitial exudative and leakage inflammation, often with the formation of hyaloid membranes in pulmonaryalveoli, and the haemorrhagic necrosis inflammation of immune organs might be pathological features of SARS. According to the structures, diameter and location of the virus like particles found in this case, combined with the pathological changes, we should consider that those virus like particles might be a new kind of coronavirus, and this kind of virus might be the main causative agent of SARS. However, the chlamydia like particles frequently observed in extra lung organs also suggested the potential new kind of coronavirus might be coexist and synergicallly cause SARS. Our findings in this study provide several evidences for SARS clinical therapy such as application of corticosteroid and enhancement of immune ability and combination of anti virus and anti chlamydium drugs.

Objective To study the pathological changes in organs remote from the lung in SARS patient. Methods The pathological changes in extra lung organs and potential coronavirus infection were studied by using light and electron microscopic examinations as well as special virus inclusion stains in the tissues obtained from an autopsy of a patient who died of SARS. Results Besides the lesions in the lung, pathological changes were found also in the central nervous system (CNS), including the cerebrum, cerebellum, thalamus, pons, and medulla oblongata, such as widening of the Virchow Robin′s space, infiltration of a few lymphocytes and macrophages in the parenchyma, vasodilatation and congestion. However, no significant neuron degeneration or necrosis was identified. Vasodilatation in the lamina propria of mucosa and submucosa of the digestive tract with some lymphocytes infiltration, and epithelial nuclear vacuolation, and occasional apoptosis were observed in the mucosal epithelial and glandular cells, as well as focal hemorrhage in segments of the small intestine. Mesenchymal edema and infiltration of a few lymphocytes in the pancreas were noted. Very mild lymphocyte infiltration, but no viral inclusions, was found in the convoluted seminiferous tubules of the testis. The patient who died of SARS was proved to have arteriosclerosis of the coronary arteries, and coronavirul particles were identified in the blood vessels under electron microscopic examination, however no coronavirul particles were found in the brain or the testis of the patient. Conclusion There were mild hypoxic changes in the tissue of CNS in the patient with severe SARS without invasion of the virus. It was confirmed that there were coronavirul particles in the blood of the patient at the acute stage of SARS. Since the patient who succumbed to the disease had a history of coronary arteriosclerosis, it was inferred that cardiovascular disease might be a contributory factor of mortality in this patient with severe SARS.

Objective To explore the status of immune responses in the lungs and the changes in lymphocyte subgroups in the immune organs in a patient having been suffered from severe acute respiratory syndrome (SARS). Methods The distribution and number of lymphocyte subgroups in the lungs and immune organs from an autopsy case of SARS were analyzed by using immunochemical staining with an array of monoclonal antibodies including CD3, CD4, CD8, CD20, CD57, CD68, S-100 and HLA-DR. Healthy spleen and lymph nodes were used as normal controls. Results CD8 + T lymphocytes constituted the major component of infiltration of inflammatory cells in the pulmonary interstitium. A semi-quantitative analysis of lymphocyte subgroups revealed that the percentage of CD3 +, CD4 +, CD8 + or CD20 + lymphocyte in a total of 31 thoracic lymph nodes of the SARS case were decreased by 74.2%, 67.7%, 74.2%, and 83.9%, respectively, compared with healthy controls. However, the percentages of lymphocyte subgroups in the celiac lymph nodes were less decreased than those in thoracic lymph nodes. The numbers of CD20 + , CD3 +, CD4 + and CD8 + lymphocytes were also decreased. CD20 + lymphocyte were notably decreased in the spleen, while CD57 +, CD68 +, S-100 + and HLA-DR + cells were increased relatively in the lymph nodes and spleen. Conclusions The results suggested that cellular immune responses were predominant in the lung of SARS patient, and it might play an important role in getting rid of coronaviruses in the infected cells and inducing immune mediated injuries to the lungs. There might be a decrease in number and imbalance in various degrees in the proportion of lymphocyte subgroups in the immune organs of the patients with severe SARS, and these changes might have a tendency to be more remarkable in lymphatic tissue situated closer to the lungs.

Objective To explore the target cells of SARS coronavirus infection in vivo and to provide the evidence of multi organ injuries produced by SARS coronavirus infection. Methods Three biotin labeling oligonucleotide probes were synthesized according to the published gene sequence of SARS coronavirus. The location, distributtion and quantity of SARS coronavirus in 2 autopsy cases of SARS were studied by in situ hybridization and electron microscopic examination. Results SARS coronavirus particles were identified in multiple organs. In lungs, SARS coronaviruses were located predominantly in the cytoplasm of bronchiolar and alveolar epithelial cells, in a part of macrophages and endothelial cells as well as a few infiltrated lymphocytes. In situ hybridization showed that in target cells SARS coronavirus distribution presented a cytoplasmic or inclusive pattern, and the mean number of positive cells in the pulmonary tissue was 80?25 per 200? field. Electron microscopic examination showed that the coronaviral particles were 100～150 nm in diameter, with low density electron cores with halo or garland envelopes. About 15% of renal tubular epithelial cells harbored SARS coronavirus, and a few parenchymal cells and sinusoid capillary endothelial cells of adrenal glands were hybridization positive. In the gastro intestinal tract, SARS coronaviruses were seen in the cytoplasm of mucosal and crypt epithelial cells, mostly in 2/3 of superficial mocosa. Under both electron microscopy and in situ hybridization observation, SARS coronaviruses were found focally distributed in some cardiomyocytes. The SARS coronavirus positive particles were also noted in macrophages/histocytes, sinusoid endothelial cells, as well as a few lymphocytes in thoracic and celiac lymph nodes. In addition, coronavirus particles were also seen in a few testicular epithelial cells and Leydig's cells. Conclusion SARS coronavirus could attack multiple target cells, implicating that SARS might cause multi organ damages, with lungs as the predominant organ of injury.

Objective To explore the significance of cellular apoptosis induced by severe acute respiratory syndrome cronovirus (SARS-CoV) in the pathogenesis of SARS. Methods TdT-mediated biotinylated-dUTP nick end ladelling method (TUNEL), and the double immunochemical staining with cytokeratin and CD3, CD8, CD20 and CD68 monoclonal antibodies were used to study the cellular apoptosis in tissue specimens from six patients who died from SARS. Meanwhile, the expression of Fas, Fas ligand (FasL), P53 and Bcl-2 proteins was detected by immunohistochemical method. Results The number of cellular apoptosis was obviously increased in multiple organs from the six patients died from SARS. The cellular apoptosis occurred predominantly in cytokeratin-positive pneumoncytes, terminal bronchiolar epithelium, CD3 + and CD8 + lymphocytes, as well as a part of CD20 + lymphocytes and CD68 + macrophages. Fas protein was mainly expressed in the infiltrated mononuclear cells, while FasL was chiefly expressed in SARS-CoV target cells, especially in the apoptotic cells. In the lung and immune organs, down-regulation of P53 and Bcl-2 expression was found. Conclusion The occurrence of increased and rapid cell apoptosis induced by SARS-CoV might be the main cause of the injuries to the lung and immune system. That the activated lymphocytes which expressed Fas and FasL attack SARS-CoV target cells might be the underlying mechanism of cell apoptosis in SARS. Down-expression of Bcl-2 and P53 proteins might also participate in cell apoptosis induced by SARS-CoV.

Objective To explore the clinical pathological features of Wilson's disease and the pathogenesis of liver fibrosis. Methods The clinical data and liver biopsy specimens obtained from 48 children with Wilson's disease were analysed. The pathological changes were studied with light microscopy, electron microscopy, combined with rhodanine and rubeanic acid for copper staining, Gordon-Sweet's staining for reticular fibers and Masson's staining for collagen fibers. Meanwhile, immunohistochemistry was used to investigate the expression of tissue inhibitors of metalloproteinase (TIMP) -1 and TIMP-2. The apoptosis of activated hepatic stellate cells (HSC) in liver tissues was illustrated with in situ end labeling (ISEL)(TUNEL POD method) and ?-SMA double staining. Results In all the cases, the mean onset age was 10.0?3.8 years, and the positive rates of family history, Kayser-Fleischer’s ring and decreased serum ceruloplasmin level were 29.2%, 68.8% and 93.0%, respectively. The levels of serum ALT, AST, ALP and ?-Glo were 3.6, 3.0, 2.7 and 2.0 fold of normal cutoff values. The major pathological changes in childhood patients with Wilson’s disease presented various chronic inflammatory changes in hepatic acini and portal tracts, interface hepatitis, focal or diffuse vesicular/ microvesicular steatosis, with large and irregular apoptotic bodies, Mallory's bodies, glycogenated nuclei, and eosinophilc granular hepatocytes. Among all the cases, 77.0% of liver specimens were positive for rhodanine and rubeanic acid staining for copper in hepatocyts, especially in the zone I of acinus. Ultrastructural observation showed swollen and unusual giant mitochondria, increased lysosomes and vesicular inclusions in hepatocytes. The incidence of hepatic fibrosis was 100%, presenting expanded portal tracts in the early, fibrotic septa in the moderate and cirrhosis in the late stage. The extent of TIMP-1 and TIMP-2 expression and number of activated HSC were increased in various degrees in all the liver specimens, while apoptotic HSCs were obviously decreased in the majority of cases. Conclusions The clinical and pathological changes of children with Wilson’s disease are varied and relatively obscure, and liver fibrosis appears early and progressive. Excessive activation and proliferation of HSC stimulated by liver injury and inflammation due to copper deposition and the decrease in activity of matrix degradation enzymes might be the important mechanisms underlying the occurrence and progression of hepatic fibrosis in Wilson's disease.