Calcium has been implicated as a primary pathogenetic mediator of cellular injury under conditions of oxygen and substrate deprivation in the kidney aswell as other tissues. It has been proposed that calcium entry from the external medium increases intracellular free calcium to toxic levels there-by converting cellular injury from a potentially reversible to an irreversible state. Cellular calcium homeostasis is a fundamental prerequisite for many coordinate cellular functions. The regulation of cellular Ca++ -metabolism is maintained mainly by Ca++ -ATPase. The purpose of the present study was to determine the effects of ischemia on the subcellular specific activity of Ca++ -ATPase of renal tissues in the rabbits. 1. In both normal cortical and medullary tissues, the specific activities Ca++ -ATPase were higher in mitochondria than in cytosol. 2. In cortical tissue, the specific activities of mitochondrial Ca++ -ATPase were increased with duration of ischemic time under normothermia. But the specific activities were decreased comparing to those of control under hypothermia, and the activities were not different with those of control in Verapamil treated group. 3. The specific activities of Ca++ -ATPase of cortical cytosol were increased as in mitochondria under normothermia. And the activities were similar with those of control in hypothermic and Verapamil treated groups. 4. In medullary tissues, the specific activities of mitochondrial Ca++ -ATPase were increased with duration of ischemic time under normothermia. In Verapamil treated group, the activities were decreased comparing to those of control. And there was no difference of activities between hypothermia and control groups. 5. As in cortical tissues, the specific activities of Ca++ -ATPase of medullary, cytosol were increased in normothermic group. And the activities were similar with those of control in hypothermic and Verapamil treated groups. Above results suggested that calcium contents of mitochondria and cytosol were increased with duration of ischemic time under normothermia. Hypothermia and Verapamil treatment seemed to be beneficial effects in prevention of calcium accumulation in the renal tissues.
Adenosine Triphosphatases* ; Calcium ; Cytosol ; Homeostasis ; Hypothermia ; Ischemia* ; Kidney ; Mitochondria ; Oxygen ; Rabbits* ; Subcellular Fractions* ; Verapamil

PURPOSE: To Study the possible mechanisms of change of thiosulfate sulfurtransferase (TST) activity in cholestatic rat liver and serum. METHODS: Rats were divided into seven groups: those receiving a sham operation (Sham group), with a bile duct obstruction (BDO) alone (BDO group), with a BDO plus taurocholic acid (TCA) injection (BDO plus TCA group), with a BDO plus tauroursodeoxycholic acid (TUDCA) injection (BDO plus TUDCA group), a choledocho-caval shunt (CCS) operation (CCS groups), a CCS operation plus TCA injection (CCS plus TCA group) and a CCS operation plus TUDCA injection (CCS plus TUDCA group). The TST activities in the serum and in the hepatic subcellular fractions isolated from above experimental rats were determined. The Km and Vmax values of this hepatic enzyme were measured. RESULTS: The liver cytosolic, mitochondrial and microsomal TSTs activities, as well as the TST Vmax values were found to be significantly decreased in the BDO plus TCA and BDO groups compared to the control group. The activity and Vmax value of the liver cytosolic TST were also found to be significantly decreased in the CCS plus TCA group. Conversely, there was no variation in the Km values of the hepatic enzymes in any of the above experimental groups. The serum TST activities in the CCS plus TCA and BDO plus TCA groups, were significantly increased compared with the control, CCS and BDO groups. However, the serum and hepatic enzyme activities were unchanged in both the CCS plus TUDCA and BDO plus TUDCA groups. CONCLUSION: The above results indicate that TCA represses the biosynthesis of TST in the liver. Also, the elevated TST activity in the serum is most likely due to an increase in the permeability of hepatocytes membrane upon TCA mediated liver cell necrosis.
Administration, Intravenous* ; Animals ; Cholestasis ; Cytosol ; Hepatocytes ; Liver* ; Membranes ; Necrosis ; Permeability ; Rats* ; Subcellular Fractions ; Taurocholic Acid* ; Thiosulfate Sulfurtransferase*

PURPOSE: To Study the possible mechanisms of change of thiosulfate sulfurtransferase (TST) activity in cholestatic rat liver and serum. METHODS: Rats were divided into seven groups: those receiving a sham operation (Sham group), with a bile duct obstruction (BDO) alone (BDO group), with a BDO plus taurocholic acid (TCA) injection (BDO plus TCA group), with a BDO plus tauroursodeoxycholic acid (TUDCA) injection (BDO plus TUDCA group), a choledocho-caval shunt (CCS) operation (CCS groups), a CCS operation plus TCA injection (CCS plus TCA group) and a CCS operation plus TUDCA injection (CCS plus TUDCA group). The TST activities in the serum and in the hepatic subcellular fractions isolated from above experimental rats were determined. The Km and Vmax values of this hepatic enzyme were measured. RESULTS: The liver cytosolic, mitochondrial and microsomal TSTs activities, as well as the TST Vmax values were found to be significantly decreased in the BDO plus TCA and BDO groups compared to the control group. The activity and Vmax value of the liver cytosolic TST were also found to be significantly decreased in the CCS plus TCA group. Conversely, there was no variation in the Km values of the hepatic enzymes in any of the above experimental groups. The serum TST activities in the CCS plus TCA and BDO plus TCA groups, were significantly increased compared with the control, CCS and BDO groups. However, the serum and hepatic enzyme activities were unchanged in both the CCS plus TUDCA and BDO plus TUDCA groups. CONCLUSION: The above results indicate that TCA represses the biosynthesis of TST in the liver. Also, the elevated TST activity in the serum is most likely due to an increase in the permeability of hepatocytes membrane upon TCA mediated liver cell necrosis.
Administration, Intravenous* ; Animals ; Cholestasis ; Cytosol ; Hepatocytes ; Liver* ; Membranes ; Necrosis ; Permeability ; Rats* ; Subcellular Fractions ; Taurocholic Acid* ; Thiosulfate Sulfurtransferase*

We have investigated the effect of cholestasis on the hepatic thiosulfate sulfurtransferase (rhodanese) and UDP-glucuronosyltransferase (UDP-GT) activities in rats. Rhodanese activities in the liver cytosol, mitochondria and microsomal fractions as well as in the rat serum, and UDP-GT activity in the microsome have been investigated for a period of 42 days after common bile duct (CBD) ligation. The cytosolic rhodanese activity showed a significant decrease between the first through the 42nd day, and the mitochondrial activity showed a significant decrease between the 7th through the 42nd day after CBD ligation compared to the activities from the sham operated control, respectively. In the case of microsomal preparation, both rhodanese and UDP-GT also showed significant decrease in their activities after the ligation for the former enzyme between the 14th and the 42nd days, and for the latter enzyme between the third and 42nd days, respectively. On the other hand, the serum rhodanese activity increased markedly soon after the ligation, exhibiting the peak activity after 1 day of CBD ligation with about 4.6-fold increment. The activity subsequently decreased gradually reaching to the control level at the 42nd day post-ligation. Enzyme kinetic parameters of hepatic rhodanese and UDP-GT were analyzed using sodium thiosulfate and p-nitrophenol as substrates, respectively, with the preparations from the 28th day post-ligation. The results indicated that although the K-m values of these enzymes were about the same as the sham-operated control, the V-max values of the both enzymes decreased significantly. These results, therefore, suggest that the biosynthesis of rhodanese and UDP-GT have been reduced in response to cholestasis, and that the elevation of rhodanese activity in the serum is most likely due to leakage from the liver subsequent to CBD ligation.
Animals ; Cholestasis ; Common Bile Duct* ; Cytosol ; Hand ; Ligation* ; Liver* ; Microsomes ; Mitochondria ; Rats* ; Sodium ; Thiosulfate Sulfurtransferase*

PURPOSE: The possible mechanisms of increased thiol me thyltransferase (TMT) activity in cholestatic rat livers and serum were studied. METHODS: Rats were divided into seven groups: rats receiv ing a sham operation, rats with a bile duct obstruction (BDO) alone (BDO group), rats with BDO plus taurocholic acid (TCA) injection (BDO plus TCA group), rats with BDO plus tauroursodeoxycholic acid (TUDCA) injection (BDO plus TUDCA group), rats receiving a choledoco-caval shunt (CCS) operation (CCS groups), rats receiving a CCS operation plus TCA Injection (CCS plus TCA group), and rats receiving a CCS operation plus TUDCA injection (CCS plus TUDCA group). The TMT activities in the serum and in the hepatic subcellular fractions isolated from these experimental rats were determined. The values of Km and Vmax in this he patic enzyme were measured. RESULTS: The activities of liver mitochondrial and microsomal TMTs as well as the Vmax values of TMT were found to be increased significantly in both the CCS plus TCA and the BDO plus TCA groups, compared with the CCS and BDO groups. On the other hand, the Km values of hepatic subcellular TMT were the same in all experimental groups. The serum TMT activity increased significantly in both the CCS plus TCA and the BDO plus TCA groups, compared with the control, CCS and BDO groups. However, these serum and hepatic enzyme activities were the same in the CCS plus TUDCA and the BDO plus TUDCA groups. CONCLUSION: The above results suggest that TCA stimulates the biosynthesis of TMT in the liver. Also, the elevated TMT activity in the serum is thought to be caused by an increase in membrane permeability of hepatocytes from liver cell necrosis caused by TCA.
Administration, Intravenous* ; Animals ; Cholestasis ; Hand ; Hepatocytes ; Liver ; Membranes ; Necrosis ; Permeability ; Rats* ; Subcellular Fractions ; Taurocholic Acid*

PURPOSE: The possible mechanisms of increased arylamine N-methyl- transferase (AMT) activity in cholestatic rat livers and serum were studied. METHODS: Rats were divided into eight groups: rats receiving a sham operation, rats with a bile duct obstruction (BDO) alone (BDO group), rats with a BDO plus taurocholic acid (TCA) injection (BDO plus TCA group), rats with a BDO plus tauroursode oxycholic acid (TUDCA) injection (BDO plus TUDCA group), rats receiving a choledocho-caval shunt (CCS) operation (CCS groups), rats receiving a CCS operation plus TCA injection (CCS plus TCA group), and rats receiving a CCS operation plus TUDCA injection (CCS plus TUDCA group). The AMT activities in the serum and in the hepatic subcellular fractions isolated from the above experimental rats were determined. The values of Km and Vmax in this hepatic enzyme were measured. RESULTS: The activities of liver mitochondrial and microsomal AMTs as well as the Vmax values of AMT, were found to be increased significantly in both the CCS plus TCA group and the BDO plus TCA group compared with the CCS and BDO groups. On the other hand, the values of Km of hepatic subcellular AMT was the same in all experimental groups. The serum AMT activity increased significantly in both the CCS plus TCA group and the BDO plus TCA group compared with control the CCS and BDO group. However, these serum and hepatic enzyme activities were the same in both the CCS plus TUDCA group and the BDO plus TUDCA group. CONCLUSION: The above results suggest that TCA stimulates the biosynthesis of AMT in the liver. Also, the elevated AMT activity in the serum is thought to be caused by an increase in the membrane permeability of hepatocytes from liver cell necrosis caused by TCA.
Animals ; Cholestasis ; Cholestasis, Extrahepatic ; Hand ; Hepatocytes ; Liver ; Membranes ; Necrosis ; Permeability ; Rats* ; Subcellular Fractions ; Taurocholic Acid* ; Transferases

OBJECTIVETo develop a method for extracting cytoskeletons and cytoskeleton-associated proteins for proteomic analysis.

METHODSA subcellular sequential proteome extraction method was exploited. The extraction procedure was optimized and controlled according to observed cell morphology changes and one- and two-dimensional electrophoresis images. The extraction efficiency and selectivity were evaluated by Western blotting and mass spectrometry.

RESULTSFour extracted fractions clearly displayed distinct patterns. Western blotting detected the fraction-marker proteins FAK, integrin-β1, histone H1 and cytokeratin 19 only in their expected fractions. About 90% of the protein spots in the cytoskeleton fraction were identified by mass spectrometry as cytoskeleton and/or its associated proteins.

CONCLUSIONThe subcellular proteome sequential fractionation method facilitates the detection of proteins of low abundance and shows a high reproducibility and selectivity, and thus can serve as an ideal pre-fractionation method prior to two-dimensional electrophoresis.

Cytoskeleton ; chemistry ; Electrophoresis, Gel, Two-Dimensional ; methods ; Humans ; Proteome ; analysis ; Proteomics ; methods ; Subcellular Fractions

In order to provide a theoretical basis for better understanding the function and properties of proteins, we proposed a simple and effective feature extraction method for protein sequences to determine the subcellular localization of proteins. First, we introduced sparse coding combined with the information of amino acid composition to extract the feature values of protein sequences. Then the multilayer pooling integration was performed according to different sizes of dictionaries. Finally, the extracted feature values were sent into the support vector machine to test the effectiveness of our model. The success rates in data set ZD98, CH317 and Gram1253 were 95.9%, 93.4% and 94.7%, respectively as verified by the Jackknife test. Experiments showed that our method based on multilayer sparse coding can remarkably improve the accuracy of the prediction of protein subcellular localization.
Algorithms ; Amino Acid Sequence ; Computational Biology ; Protein Transport ; Proteins ; Subcellular Fractions ; Support Vector Machine

This study was designed to investigate the effects of pyroligneous liquor on oxygen radicals and their scavenger enzy-mes in the liver of Cri/Bgi CD rats (7 rats per group ). Male rats were fed a basic diet prepared in our Lab., PL-0 (Control ), PL-1, PL-25, PL-50 and PL-75 groups were prepared to be 0%, 1%, 25%, 50% and 75%with distilled water using pyrolig-neous liquor (35% of Choa Co. Ltd.), and were administrated orally for 8 weeks. Superoxide radical contents in liver mi-tochondria and microsomes were significantly decreased to 12-14%, 11-15%, respectively, in these PL-25 and PL-50 groups compared with the control group. Hydroxyl radical content in mitochondria and microsomes were markedly decreased to 12-20% and 17%, respectively, in these PL-25 and PL-50% groups compared with the control group. Hydro-gen peroxide content in mitochondria and microsomes were significantly decreased about 15-12% and 22-20% in liver of PL-25 and PL-50 groups compared with the control group. Mn-SOD and Cu/Zn-SOD activities in liver of PL-25 and PL-50 groups were remarkably increased to 15-25%, 11-16%, respectively, compared with the control group. GPx activities in mitochondria and microsomes were significantly increased in the liver of PL-25 and PL-50 groups compared with the control group. CAT activities in mitochondria and cytosol were significantly increased to 12-14%, 15-27%, respectively, in the liver of PL-25 and PL-50 groups compared with the control group. These results suggest that long term administra-tion orally of 25 and 50% pyroligneous liquor may effectively inhibit the formation of oxygen free radicals, and also scavenger enzyme activities significantly increase through the administration orally.
Animals ; Cats ; Cytosol ; Diet ; Free Radicals ; Humans ; Hydroxyl Radical ; Liver* ; Male ; Microsomes ; Mitochondria ; Oxygen* ; Rats* ; Reactive Oxygen Species* ; Superoxide Dismutase ; Superoxides ; Water

OBJECTIVETo investigate the effects of Hedan Tablet () on serum lipid profile, proprotein convertase subtilisin/kexin type 9 (PSCK9) and high-density lipoprotein (HDL) subfractions in patients with hyperlipidemia.

METHODSThirty-seven patients with hyperlipidemia were randomized to treatment with Hedan Tablet 4.38 g/day as Hedan group (18 cases) or placebo (19 cases) as control group for 8 weeks. The lipid profile, PCSK9 and HDL subfractions were determined at day 0 and week 8 in both groups respectively.

RESULTSHedan treatment for 8 weeks mildly decreased serum low-density lipoprotein cholesterol (LDL-C) levels, while no changes were found in total cholesterol (TC), triglycerides (TG) and PCSK9 concentrations. Furthermore, Hedan treatment increased the concentration of large high-density lipoprotein cholesterol (HDL-C) and the percentage of large HDL subfraction, while decreased the concentration of small HDL-C and the percentage of small HDL subfraction without changing serum HDL-C levels in patients with hyperlipidemia.

CONCLUSIONHedan treatment of 4.38 g per day for 8 weeks could confer a favorable effects on serum LDL-C concentration as well as HDL subfractions.

Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Hyperlipidemias ; blood ; drug therapy ; enzymology ; Lipoproteins, HDL ; blood ; Male ; Middle Aged ; Proprotein Convertase 9 ; metabolism ; Subcellular Fractions ; metabolism