ABSTRACT
Objective To localize endogenous alkaline phesphatase in human neutrophils with ELF(enzyme-labeled fluorescence)-97 phosphate,which yields an intensely fluorescent yellow-green precipitate at the site of enzymatic activity. Methods Neutrophils were isolated from human blood and fixed,then histochemistry with the ELF-97 phosphate was performed with the ELF-97 endogenous phosphatase detection kit.All photography was performed with a Nikon labophot fluorescence/DIC microscope. Results Fluorescent yellow-green granules,well-distributed but size-varied,were observed in neutrophils under fluorescence microscope.There were 94.102?3.133 percent rate of positive neutrophil alkaline phosphatase among 51 cases of healthy volunteers.Conclusion ELF-97 endogenous phosphatase detection kit can provide sensitive,high-resolution localization of endogenous phosphatase activity in neutrophils.
ABSTRACT
Male mice of Kunmlng strain were infected with 10 000 tachyzoites intraperitoneally. 2 hours after infection the mice were divided into 2 groups and administered 5 % of amylurn and 200 mg/kg of artemether by gavage for 8 consecutive days. The ultrastructural enzyme cytochemical studles on cytidine monophosphatase (CMP ase) and glucose-6-phosphatase (G-6-P-ase) of the parasites were carried out. CMP-ase was found scattered in the lysosomes of the parasites as well as in the macrophages. No differences were observed in the localization and intensity of CMP-ase activity between the nontreated and treated with drug parasites. G6-P-ase was found surrounding the parasite membrane and scattered in the parasitophorous vacuole in the nontreated parasites. After treatment with artemether, the intensity of G-6-P-ase activity was decreased compared with nontreated control parasites. It is suggested that artemether may exert some action on the G-6-Patase of T. gondii and thus influence the energy metabolism of the parasite.
ABSTRACT
Berbamine ( Ber. ) is a bis-benzyl-isoquinoline alkaloid isolated from the traditional Chinese medical herb, Berberis poiretis, and has Vide pharmacological effects. Clinical studies have demonstrated that Ber. had an effect of ingreasing the number of leukocytes in various leukocytopenia. But, up till now it has not been reported that Ber. take effect on enzymic metablism of inner cell and isoenzyme in the process of increasing leukocyte. The experiment observed the mice dynamic change of LDH in leukocyte and LDH isoenzyme of serum by cytochemistry and plane polyacrylamide gel electrophoresis tech-nigue. The result indicates that LDH enzyme reaction of granulocyte have increased in Ber. group and intensity of enzyme reaction has a positive interralation with the time of administrations. All specimen show 5 zones after LDH isoenzyme pattern analyse. The proportion of LDH-5 was changed and LDH-5 sub-band was detected in control and combination group. The experiment indicated that Ber. can affeet enzymic metablism of inner cell in the process of increasing leukocyte. There was simultaneously with a change in ratio of LDH isoenzyme pattern.
ABSTRACT
The tadpole extract without large molecular protein(T-871) was prepared from dry tadpoles. This extract and RPMI1640 medium were mixed in the ratio of 1:50(V/V) to form T-871 medium. HeLa cells were cultured in the T-871 medium in order to study the possible mechanism of HeLa cell differentiation induced by tadpole extract. We found that there was a decreased acitivity of LDH which may be released through HeLa cell membrane in the 2 day's cultured T-871 medium. After 3 day's culture in the T-871 medium we found that on the HeLa cell membranes, Na-K-ATPase activity reduced and the content of Con A receptors increased. When these cells were analyzed by flow cytometry(FACS-420), the results indicated that HeLa cells accumulated in G_2 and M phases of the cell cycle and the influence on DNA and RNA content of the HeLa cells was insignificant. HeLa cells cultured by RPMI 1640 medium or T-871 medium for 3 days were inoculated into the dorsal hypodermis of 12 nude mice, respectively. Then after 24 hours this extract was injected to each nude mouse at dose of 0.3 ml each day. Our study showed that T-871 could result in decrease of tumor formation such as delay of tumor nodule appearance and reduce of tumor weight. This study suggested that the changes of the cell membrane may be caused by the tadpole extract. It may also reduce malignancy of the HeLa cells.
ABSTRACT
The selective toxicity of the mitochondria-specific cationic fluorescent dye rhodamine 123 (Rh-123) on HeLa cells in culture was studied. In this report, we demonstrate that with continuous exposure, Rh-123 markedly inhibited the growth of HeLa cells but had little effect on normal human kidney fibroblasts. With continuously exposed to Rh-123, the growth rate, colony forming ability, and mitotic index of HeLa cells were decreased. The mechanism of toxicity of Rh-123 on HeLa ceils was investigated by EM and enzyme cytochemistry stain. The mitochondria of carcinoma cells were the main targets for the inhibitory action of Rh-123, since they selectively accumulated the dye. At the dosage of Rh-123 which was toxic to HeLa cells, the structure and function of mitochondria were disrupted, as the mitochondriarelated enzymes, i. e., ATPase, LDH, and SDH were inhibited. The possible mechanism of the action of Rh-123 on HeLa cells is briefly discussed.