ABSTRACT
<p><b>OBJECTIVE</b>To explore cell culture techniques for amplification of oval cells with preservation simultaneously of the stem cell characteristics.</p><p><b>METHODS</b>Oval cell line OC3 was cultured in RPMI 1640 supplemented with 15% fetal bovine serum and 20 µg/L EGF. Cells were harvested every 5 passages and were examined with biomarkers including OV-6, c-kit, gamma-glutamyl transpeptidase, placental form of glutathione-S-transferase (GST-P), pyruvate kinase M₂, pyruvate kinase L and albumin using techniques including RT-PCR, immunocytochemistry, and enzymo-cytochemistry.</p><p><b>RESULTS</b>OC3 cell lines could be amplified abundantly in-vitro associating with expression of infant liver cell markers at various level, including OV-6, c-kit, gamma-glutamyl transpeptidase, GST-P, pyruvate kinase M₂, but no expression of mature hepatocyte markers detected including pyruvate kinase L and albumin.</p><p><b>CONCLUSIONS</b>Amplification of OC3 cells with preservation of the stem cell phenotype and high proliferation index can be achieved up to the 79(th) passages by culturing in RPMI 1640 supplemented with 15% fetal bovine serum and 20 µg/L EGF.</p>
Subject(s)
Animals , Rats , Antigens, Differentiation , Metabolism , Cell Culture Techniques , Cell Differentiation , Cell Line , Culture Media , Glutathione Transferase , Metabolism , Hepatocytes , Cell Biology , Metabolism , Liver , Cell Biology , Phenotype , Proto-Oncogene Proteins c-kit , Metabolism , Pyruvate Kinase , Metabolism , Rats, Sprague-Dawley , Stem Cells , Cell Biology , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the clinical manifestation and pathological features of chronic rejection (CR) and the management of CR after orthotopic liver transplantation (OLT).</p><p><b>METHODS</b>From January 2004 to December 2006, there were 516 patients who had undergone OLT. All the clinical and pathological data were collected and retrospectively studied. Clinical manifestation, histopathological feature, diagnosis and anti-rejection treatment of CR were summarized and analyzed.</p><p><b>RESULTS</b>The incidence of CR was 2.3% (12/516), including 7 cases with early phases of CR and 5 cases with late phases of CR. The main pathological changes of CR were the vanishing bile duct syndrome and obliterative arteriopathy;and the early stage of CR were the damage of inter lobular bile duct, necrotic inflammation in central lobule, and inflammatory cells infiltration in portal area. Among 12 patients with CR, 7 cases with early CR were reversed by methylprednisolone (MP) pulse treatment and adjusting immunosuppressant dose, including 2 cases of whom were prescribed OKT3 treatment and 2 cases treated by ATG, and 5 cases with late CR underwent liver retransplantation (re-LT). Two patients died from infection, 1 case died from multiple organ failure in perioperative period after re-LT, another 2 cases were cured by re-LT, and the CR related mortality was 25.0% (3/12).</p><p><b>CONCLUSIONS</b>Chronic rejection following OLT is lack of typical clinical manifestation and pathological features, and the pathological changes can overlap and coexist. Post-transplant liver biopsy and graft specimen after re-LT is still "gold standard" to CR diagnosis. Some of early CRs can be reversed by early diagnosis and early treatment; for late CR recipient, re-LT should be considered.</p>
Subject(s)
Adolescent , Adult , Aged , Child , Female , Humans , Male , Middle Aged , Young Adult , Chronic Disease , Graft Rejection , Diagnosis , Pathology , Therapeutics , Liver Transplantation , Retrospective StudiesABSTRACT
<p><b>BACKGROUND</b>Acute rejection is one of the most important factors for prognosis following liver transplantation. With the use of potent immunosuppressants, acute rejection does not always present typical manifestations. Moreover, other complications often occur concomitantly after liver transplantation, which makes early diagnosis of acute rejection more difficult. Acute rejection is best diagnosed by liver biopsy. Differentiation of clinical manifestations and pathological features plays an important role in achieving individualized immunosuppressive treatment and prolonging long term survival of patients given orthotopic liver transplants.</p><p><b>METHODS</b>From January 2004 to December 2006, 516 orthotopic liver transplantations were performed at the First Affiliated Hospital, Sun Yat-sen University. For patients who suffered acute rejection, clinical manifestations, histopathological features, diagnosis and anti-rejection treatment were summarized and analyzed.</p><p><b>RESULTS</b>In 86 cases (16.7%), of the 516 recipients, 106 episodes of acute rejection occurred, which included 9 with histopathological borderline changes, 36 Banff I rejections, 48 Banff II and 13 Banff III. Among these, 36 were cured by adjusting the dose of immunosuppressant and 65 were reversed by methylprednisolone pulse treatment. Five were methylprednisolone resistant, 3 of whom were given OKT3 treatment and 2 underwent liver retransplantation.</p><p><b>CONCLUSIONS</b>Due to potent immunosuppressive agents, acute rejection following an orthotopic liver transplantation lacks typical clinical manifestations and pathological features. Acute rejection is best diagnosed by liver biopsy. Designing rational individualized immunosuppressive regimen based on clinical and pathological features of acute rejection plays an important role in prolonging long term survival of patients.</p>
Subject(s)
Adolescent , Adult , Aged , Child , Female , Humans , Male , Middle Aged , Young Adult , Graft Rejection , Drug Therapy , Pathology , Immunosuppressive Agents , Therapeutic Uses , Liver Transplantation , Methylprednisolone , Therapeutic UsesABSTRACT
<p><b>OBJECTIVE</b>To characterize the biologic featrues of hepatic oval cells and their protein expression profiles during induced differentiation in vitro.</p><p><b>METHODS</b>Rat hepatic oval cells were treated with epidermal growth factor (EGF) and hepatocyte growth factor (HGF) in vitro, followed by morphological and molecular marker assessment by electromicroscopy, immunocytochemistry, RT-PCR and protein expression chip technology.</p><p><b>RESULTS</b>Ten weeks after induction, the levels of GST-P mRNA and M2-PK mRNA were significantly reduced, whereas those of ALB and CK18 were elevated. Significant variations of expression was seen in 8 protein species during the course of the induced differentiation.</p><p><b>CONCLUSION</b>Combined EGF and HGF treatment in vitro induces cell differentiation of hepatic oval cells, a process in which 8 protein species may play some regulatory roles.</p>
Subject(s)
Animals , Rats , Albumins , Metabolism , Cell Differentiation , Epidermal Growth Factor , Pharmacology , Glutathione Transferase , Genetics , Hepatocyte Growth Factor , Pharmacology , Hepatocytes , Cell Biology , Metabolism , Immunohistochemistry , Keratin-18 , Metabolism , Protein Array Analysis , Pyruvate Kinase , Genetics , RNA, Messenger , Metabolism , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
<p><b>OBJECTIVE</b>To study the effects of enhanced external counterpulsation (EECP) on the vascular morphology, and endothelial function using experimentally induced hypercholesterolemic pigs.</p><p><b>METHODS</b>Thirty five male pigs were randomly divided into three groups: 7 normal control animals, 11 hypercholesterolemic animals, and 17 hypercholesterolemic animals receiving EECP. Serum cholesterol was measured. The coronary arteries and aortas were sampled for histopathologic and ultrastructural examination. The NF-kappaB protein expression of porcine coronary arteries was investigated by immunofluorescence.</p><p><b>RESULTS</b>Compared with the normal controls, serum cholesterol levels were significantly higher in the hypercholesterolemic animals with or without EECP. The plaque/intimal area ratio of the aorta decreased significantly in animals receiving EECP [(3.33 +/- 2.40)%, versus (12.03 +/- 7.12)% in those without EECP, P < 0.05]. Lipid deposition, endothelial damage and proliferation of smooth muscle cells were less severe in animals receiving EECP than those not. Moreover, activation and expression of NF-kappaB also decreased significantly (P < 0.05) in animals receiving EECP.</p><p><b>CONCLUSIONS</b>EECP improves the morphology and function of vascular endothelium, and retards the development and progression of atherosclerosis, likely through the inhibition of NF-kappaB signaling pathway.</p>
Subject(s)
Animals , Male , Aorta, Abdominal , Metabolism , Pathology , Atherosclerosis , Blood , Metabolism , Pathology , Cholesterol , Blood , Coronary Vessels , Metabolism , Pathology , Counterpulsation , Methods , Endothelial Cells , Metabolism , Pathology , Hypercholesterolemia , Blood , Metabolism , Pathology , Lipoproteins, LDL , Blood , Microscopy, Confocal , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Muscle, Smooth, Vascular , Metabolism , Pathology , NF-kappa B , Metabolism , Random Allocation , SwineABSTRACT
Objective To report a peliosis hepatic in child and review literature and discuss.Methods Case history was inquired.Physical,labtoratory,imagement and histopathology of liver biopsy(HE staining) were examed.Results A 4-year old girl appeared dermatitis with erythema and herpes at local skin where was bit by insect before onset.The girl appeared fever,cough,then abdominal pain,hepatomegaly,pleural effusion and ascites.Lab examination revealed slight elevation of aspartate transaminase,?-glutamyltranspeptidase and alkaline phosphatase.The liver B-mode ultrasonography and CT scan revealed hepatomegaly with density heterogeneity of the parenchyma.The liver biopsy revealed many small capsule filled with blood cells.Conclusions Clinical characteristics of the disease are fever,upper abdomen pain,janundice,ascites and hepatomegaly.The diagnosis shall be combined with the pathologic biopsy of liver.