| PrefaceHepatic fibrosis is the common result of many chronic hepatic diseases. It will result in hepatic cirrhosis and descend liver functional reserve, so it has been one severe disease threatening people health.It' s important and significant to research the development process & pathogenic mechanism of hepatic fibrosis and evaluate fibrosis degree objectively. This study quantitatively detected the relationship between hepatic fibrosis and TGF 1 & FN, which was carried out through several special histologic staining of hepatic puncturing specimen and computer assisted digital image analysing system, and we hope to enrich the study methods of hepatic fibrosis biopsies and determine the dependability of meamure method with computer.Material and Method1.1 Experiment Object:57 hepatitis B virus carriers and chronic hepatitis B patients (2002. 10 ~ 2003. 12 ) , 47 males & 10 females(age 16 ~53, mean 32.5 years) ,including 9 carriers ,23 mild chronic hepatitis B patients ,20 moderate and 5 severe ones. The clinical diagnosis standard accords to<>. We got at least 1cm specimens with rapid liver puncture method, paraffin imbedded and got 4 pieces of 4 m sections. Every sections was carried on HE staining ,Masson staining, TGF 1 and FN staining respectively.1.2 Experiment Reagents: TGF l & FN reagent boxes1.3 Experiment instruments: Meta Morph/DP10/BX41 colour image analy-sing apparatus ( UIC/OLYMPUS. US/JP). OLYMPUS C3040-ADU 3J 10641 colour image photograph importing apparatus.2. Experiment Methods;2. 1 Pathological grades; In blind situation with the patients information, two pathological experts drew out two sections in four at random with HE stain and Masson stain. According with {virus hepatitis preventing project} , the sections were graded to determine the hepatic fibrosis degree. If the experts diverge in opinions , the third one would appointed as the arbiter.2.2 Experiment groups; According with the hepatic fibrosis degree these sections were separated into 5 groups ;S0(n=7) ;S1(n = 12);S2(n = 15);S3 (n = 16) ; S4(n =7). The two left sections were stained and analysed with TGF 1 & FN immunohistochemical method. TGF 1 immunohistochemical results; positive products lied in cell plasma, and the positive standard was defined as brown-yellow granules. FN immunohistochemical results; positive products lied outside the cell, which were stained brown-yellow colour. Under the 40 multiple microscope, the observed results were transferred into Meta Morph/ DP10/BX41 colour image analysing apparatus with OLYMPUS C3040-ADU colour image photograph importing apparatus. In every sections the core and four outer regions were chosen, and we measure the positive reaction area ratio in the most positive products amassed, which was called mean light density value, and we got the means. Masson stain; The collagenous fiber was stained blue. This method was to measure the summation of blue-stain pixels, and the value was divided by the total pixel value of section, so we got the area ratio of collagenous fiber. These three positive area ratio values expressed the actual degree.2.3 Statistics calculations; The quantity datum were expressed with x s. SPSS 10. 0 software and Spearman Test were used to analysis the data. We thought it' s significant when P less than 0.05.Results1. Pathologic results; The result was grade 0; 7 cases; grade 1; 12 cases; grade 2; 15 cases; grade 3; 16 cases; grade 4: 7 cases.2. TGFpl level; In the early stage of fibrosis TGFpl expression increased, and its expression increased with the degrees of fibrosis. The TGFpl expressions had notable correlation with the hepatic fibrosis degree, and the correlation coefficient was 0.902(P = 0.00) .3. FN level: In the early stage of fibrosis FN expression increased, and its expression increased with the degrees of fibrosis. But the expression decreased in hepatic cirrhosis. They had no significant correlation, and correlation coefficient was 0. 188(P<0.05).
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