In the present study, comparison have been made between two kinds of double labelling methods; single side labelling and both sides labelling. To make the different groups be comparable, all sections were continuously cut from the same specimen and all labelling processes were carried out at the same ti- me. The groups were divided as follows: A, one side anti-?-amylase labelling and one side antitrypsin labelling; B, single side labelling of anti-?-amylase and antitrypsin; C, one incubation with anti-?-amylase, followed by two incu- bations with protein A-gold (PAG) complexes of varied size, C1: 7nm and 20nm, C2: 10nm and 20nm, C3: 20nm and 7nm; D, single side labelling of anti- ?-amylase and an unrelated antiserum (antichathepsin D), applying free protein A between two labellings; E, as C, but with free protein A between two PAG incubations; F, as control. Group A and B showed that the two labelling me- thods had almost the same sensitivity.Group C indicated that the interaction of single side labelling were resulted from the combination of the second PAG with the free Fc region of the first antiserum. To decrease the interaction, it was necessary for the second PAG to be much larger than the first one.Group D de- monstrated that the interaction between the second antiserum and the first PAG was very feeble. Group E proved that free protein A could completely prevent the interaction of single side labelling method. The both sides labelling method avoids interaction, but mistakes resulting from the ultrastructural differences on two sides of the sections may happen. Which method to be selected is dependent upon what to be labelled.

Purpose　To study the correlation of integrinα4β1 and its two ligands (vascular cell adhesion molecule-1 and fibronectin)with mast cell（MC）recruitment around the rat hepatocarcinoma. Methods　Eighteen male Wistar rats with liver tumor were divided into three different groups in terms of mast cell numbers in the surroundings of liver tumor. Eight normal Wistar rats were used as control. Integrin VLA-4 expression in rat peritoneal mast cells was analyzed by indirect immunofluorescence and flow cytometry. Immunohistochemistry was also used to investigate whether endothelial cell VCAM-1 and fibronection were expressed positively. Results　There were markedly different in mast cell numbers around rat liver tumor. Mast cells express high levels of integrin α4β1 on their surfaces. And the more mast cells around liver tumor, the higher levels of integrin VLA-4. We also found that endothelial cells expressed VCAM-1 and there are a number of fibronectin deposition around rat hepatocarcinoma. Conclusions　Integrin α4β1/VCAM-1 and fibronectin play an important role in mast cell recruitment mechanism around liver tumor. The expression levels of integrin α4β1 are paralleled by mast cell accumulation in the surroundings of hepotocarcinoma.

To comparatively investigate ultrastructural characteristics and expressions of AFP (alpha-fetoprotein) and Tn (Thomsen-Friedenreich-related antigen) protein in AFP negative (AFP-) and AFP positive (AFP+) primary hepatocellular carcinoma. Fourty-three cases of AFP- and AFP+ hepatocellular carcinoma (HCC) tissues and five cases of normal liver tissues were divided into three groups: control group (normal liver tissue, n=5); AFP+ HCC group (the serum AFP level was higher than 10 ng/ml, n = 22); AFP- HCC group (the serum AFP level was lower than 10 ng/ml, n=21). The ultrastructural morphology was studied by transmission electron microscopy, the expressions of AFP and Tn protein were detected by immunohistochemistry and cell image analysis. 1. The immunohistochemical study showed that (1) the expression intensity and positive rate of Tn protein in AFP- HCC group were markedly higher than that in AFP+ HCC group (P<0.01); (2) The expression intensity of AFP in AFP- HCC group was lower than that in AFP+ HCC group (P<0.01). 2. The transmission electron microscopy demonstrated that some AFP- HCC cells linked closely with each other, others dispersed loosely just as cultured cells, the remarkable morphologic features in AFP- HCC cells were simple organelles, but they were abundant in the free polyribosomes. In AFP+ HCC group, all the HCC cells linked closely together and were rich organelles in their cytoplasm, especially the rough endoplasmic reticula. In addition, mitochondria and Golgi complex were obviously observed. (1) The AFP and Tn protein had discrepancy distribution in AFP- and AFP+ HCC tissues, Tn protein may be one of the early diagnostic indicators in AFP- HCC; (2) The synthetic locations of the AFP and Tn protein were different in hepatocarcinoma cells by ultrastructural observation.

In the present study, stereological analysing methods were used to measure the nucleus, cytoplasm and secretive granules of mast cells in connective tissue and mucosa. It was found that there existed significant difference between them in nucleus/cytoplasm ratio, volume density, size and sphericity of granules. These quantitative results might be served as the main morphological parameters of distinguishing the two types of mast cells.

To comparatively investigate ultrastructural characteristics and expressions of AFP (alpha-fetoprotein) and Tn (Thomsen-Friedenreich-related antigen) protein in AFP negative (AFP-) and AFP positive (AFP+) primary hepatocellular carcinoma. Fourty-three cases of AFP- and AFP+ hepatocellular carcinoma (HCC) tissues and five cases of normal liver tissues were divided into three groups: control group (normal liver tissue, n=5); AFP+ HCC group (the serum AFP level was higher than 10 ng/ml, n = 22); AFP- HCC group (the serum AFP level was lower than 10 ng/ml, n=21). The ultrastructural morphology was studied by transmission electron microscopy, the expressions of AFP and Tn protein were detected by immunohistochemistry and cell image analysis. 1. The immunohistochemical study showed that (1) the expression intensity and positive rate of Tn protein in AFP- HCC group were markedly higher than that in AFP+ HCC group (P<0.01); (2) The expression intensity of AFP in AFP- HCC group was lower than that in AFP+ HCC group (P<0.01). 2. The transmission electron microscopy demonstrated that some AFP- HCC cells linked closely with each other, others dispersed loosely just as cultured cells, the remarkable morphologic features in AFP- HCC cells were simple organelles, but they were abundant in the free polyribosomes. In AFP+ HCC group, all the HCC cells linked closely together and were rich organelles in their cytoplasm, especially the rough endoplasmic reticula. In addition, mitochondria and Golgi complex were obviously observed. (1) The AFP and Tn protein had discrepancy distribution in AFP- and AFP+ HCC tissues, Tn protein may be one of the early diagnostic indicators in AFP- HCC; (2) The synthetic locations of the AFP and Tn protein were different in hepatocarcinoma cells by ultrastructural observation.
Antigens, Tumor-Associated, Carbohydrate ; biosynthesis ; genetics ; Biomarkers, Tumor ; Carcinoma, Hepatocellular ; metabolism ; ultrastructure ; Female ; Humans ; Immunohistochemistry ; Liver Neoplasms ; metabolism ; ultrastructure ; Male ; Tumor Cells, Cultured ; alpha-Fetoproteins ; biosynthesis ; genetics

The effects of A-L tonic capsule on DNA content in rat experimental hepatocarcinogenesis induced by diethylnitrosamine (DENA) were observed. The experimental rats were divided into 4 groups. With exception of group D in which the rats were only administered with DENA, the rats in the groups A, B, C were previously, simultaneously and subsequently fed with A-L tonic capsule respectively while they were administered with DENA. The DNA content of all rat livers was measured using automatic image analysis technique 20 weeks after administration of DENA. The results showed that the highest and lowest DNA contents were respectively seen in the groups D and A. There was significant difference in DNA contents between the groups A or B or C and D, and also between the groups A and B or C (both P < 0.01). 4 components (4C) and > or = 5C cells were predominant in the group D, while 2C cells were the minority. The number of 2C cells in the groups A, B, C was significantly higher than that in the group D, and the number of > or = 5C cells in the groups A, B, C was markedly lower than that in the group D (P < 0.01). Also, there was very significant difference in the number of 2C and > or = 5C cells between group A and B or C (P < 0.01). It was concluded that A-L tonic capsule could effectively inhibit the increase of DNA content of hepatocytes and improve the distribution of DNA content in rat hepatocarcinogenesis, especially in group A.
Animals ; Astragalus membranaceus ; chemistry ; Capsules ; Carcinogenicity Tests ; DNA, Neoplasm ; analysis ; Diethylnitrosamine ; Drugs, Chinese Herbal ; pharmacology ; Hepatocytes ; chemistry ; Ligustrum ; chemistry ; Liver Neoplasms, Experimental ; chemically induced ; chemistry ; genetics ; Male ; Rats ; Rats, Sprague-Dawley