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Objective:To improve the early diagnosis of hepatocellular carcinoma(HCC),a reversed-phase high-performance liquid chromatography(RP-HPLC)was employed to identify the differential proteins of serum related to HCC. Methods:The sera from 20 healthy volunteers and 8 HCC patients were collected. After the depletion of six serum proteins of the highest abundance with the Multiple Affinity Removal System (MARS) technologies from Agilent,the sera were subjected to RP-HPLC.The chromatographic conditions were: analytical column:ZORBAX 300SB-C18 (250 mm?4.6 mm,5 ?m);column temperature,25℃;the elution was performed with linear gradient elution of A:0.1% trifluoroacetic acid water solution and B:0.09% trifluoroacetic acid ACN solution at a flow rate of 0.75 ml/min. The detection wavelength was 280 nm. The time procedure was:1%B was held 3 min,then the gradient ran from 1 to 100% B in 100 min, then was held for another 10 min,at last,the gradient ran from 100 to 1%B in 5 min and was held for another 5 min. Results:There was a group of differential proteins of strong hydrophobicity in the serum between healthy persons and patients. Conclusion:This method can be useful in detecting protein expression alteration resulting from the carcinogenesis and development of HCC,and newly discovered biomarkers might be an aid in the early diagnosis and therapy of HCC.
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<p><b>OBJECTIVE</b>To detect the feasibility and theoretic basis for treatment with hyperbaric oxygenation (HBO) in chronic hepatitis and to compare the changes in hepatic function, immunity, pathologic morphology, ultrastructure and HBV in hepatic tissues before and after treatment.</p><p><b>METHODS</b>Sixty cases of chronic hepatitis were randomly selected and divided into two groups: the experiment (n = 30) and control groups (n = 30). Patients in the experimental group were treated with HBO for 6 courses. Patients in the control group were treated for 60 days with the usual drugs used in the clinic. The function and bloodstream graph of liver were examined and liver biopsies were made before and after treatments. Routine paraffin sections were stained with HE and observed under the light microscope. Ultra thin slides from paraformaldehyde and glutaraldehyde fixed liver tissue were stained with lead citrate and observed with the transmission electric microscope. HBsAg and HBcAg in liver of the experimental group were detected with ABC immunohistochemistry method before and after treatment.</p><p><b>RESULTS</b>For the experimental group, ALT, SB, gamma-GT, AKP, IgG, and IgM in blood and the degeneration and necrosis of hepatocytes were remarkably decreased (P < 0.05 ), the mean contractive wave of bloodstream in liver and the bloodstream in right ramus of janitrix were remarkably increased (P < 0.05), and the swelling of mitochondria, increase of lysosomes, generation of Kupffer cells, infiltration of lymphocytes in portal area and capillary generation were all remarkably all eviated (P < 0.05) after treatment with HBO. There were significant differences between the experimental and control groups after treatment with different methods (P < 0.05). For patients in the experimental group, the fibrosis and fat-storing cells in the liver were not reduced (P > 0.05), and the expression of HBsAg and HBcAg in liver was not weakened (P < 0.05) after treatment.</p><p><b>CONCLUSIONS</b>Treatment with HBO for chronic hepatitis was effective and recommendable, but it could not reverse liver fibrosis. However, it might be able to delay or prevent the liver from fibrosis, so it might be more effective at the early and middle stages of chronic hepatitis. HBO could not inhibit the HB virus. So we consider that treatment with HBO should be simultaneous with anti HBV therapy.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Hepatitis, Chronic , Pathology , Therapeutics , Hyperbaric Oxygenation , ImmunohistochemistryABSTRACT
Objective:To develop a method for the determination of vincristini sulfa(sVCR) in tumor cells culture media and intracellular fluid by high performance liquid chromatography(HPLC).Methods:VCR can be separated on C18(4.6 mm?250 mm,5?m)column by using 0.02 mol/L potassium dihydrogen phosphate:methano(l30:70)(pH=6.6)as mobile phase with UV detector at a wavelength of 267 nm.The flow rate was 0.8 ml/min.Results:The standard curve was linear in the concentration range 12.5～160.0?g/m(lr=0.997 4, n=7)and 0.125～12.5?g/m(lr=0.998 5,n=6).The average extracting recovery and average recovery of VCR in tumor cells culture media, RSD of intra-day and inter-day was 73.84%,99.83%,3.14%and 3.99%respectively.The RSD of intra-day and inter-day in intracellular fluid was 2.15%and 1.77%respectively.The detection limit was 0.04?g/ml.Conclusion:This method is simple,rapid,accurate and selective for the determination of VCR.