To investigate the earlier cellular alterations(Glucose-6-Pase activity and morphologic features) caused by a hepatotoxin, thioacetamide (TAA), a single dose of the agent (200mg per kg of body weight) was given intraperitoneally to mice, which were sacrificed at intervals of 4, 8 or 16 hours after corresponding treatments. For histochemical study of glucose-6-phosphatase (G6Pase) activity, unfixed frozen sections were incubation of the Wachstein and Meisel medium and stained. The smallest pieces of liver tissue were fixed in glutaraldehyde and osmic acid, and stained by the routine electron-microscopic techniques. Some pieces of liver were fixed in 10% formalin, embedded in paraffin, and stained with hematoxylin and eosin. There was a rapid and progressive loss of G6Pase activity, in an orderly time sequence, in the experimental group. There were also morphologic changes: loss of cytoplasmic basophilia, cell infiltration and necrosis in the centrilobular and intermediate zones, and an increase of sER, small vesicles and ribosomes in the cytoplasm of hepatocytes, the marked changes of nuclei and nucleoli, and a slight increase of lipid droplets in the cytoplasm at 16 hours after intoxication. The correlation between these cellular alterations was discussed in view of mechanisms in the hepatotoxic action.
Acetamides/adverse effects* ; Animal ; Glucose-6-Phosphatase/metabolism* ; Liver/drug effects* ; Liver/enzymology ; Liver/ultrastructure ; Male ; Mice ; Thioacetamide/adverse effects* ; MH - ; Substances: ; Acetamides ; Thioacetamide ; Glucose-6-Phosphatase

To investigate the earlier cellular alterations(Glucose-6-Pase activity and morphologic features) caused by a hepatotoxin, thioacetamide (TAA), a single dose of the agent (200mg per kg of body weight) was given intraperitoneally to mice, which were sacrificed at intervals of 4, 8 or 16 hours after corresponding treatments. For histochemical study of glucose-6-phosphatase (G6Pase) activity, unfixed frozen sections were incubation of the Wachstein and Meisel medium and stained. The smallest pieces of liver tissue were fixed in glutaraldehyde and osmic acid, and stained by the routine electron-microscopic techniques. Some pieces of liver were fixed in 10% formalin, embedded in paraffin, and stained with hematoxylin and eosin. There was a rapid and progressive loss of G6Pase activity, in an orderly time sequence, in the experimental group. There were also morphologic changes: loss of cytoplasmic basophilia, cell infiltration and necrosis in the centrilobular and intermediate zones, and an increase of sER, small vesicles and ribosomes in the cytoplasm of hepatocytes, the marked changes of nuclei and nucleoli, and a slight increase of lipid droplets in the cytoplasm at 16 hours after intoxication. The correlation between these cellular alterations was discussed in view of mechanisms in the hepatotoxic action.
Acetamides/adverse effects* ; Animal ; Glucose-6-Phosphatase/metabolism* ; Liver/drug effects* ; Liver/enzymology ; Liver/ultrastructure ; Male ; Mice ; Thioacetamide/adverse effects* ; MH - ; Substances: ; Acetamides ; Thioacetamide ; Glucose-6-Phosphatase

It is interesting and in important to study histochemical changes of glucose-6-phosphatase (G-6-Pase) activity by electron microscopy in order to promote the knowledge needed for diagnosis and prognosis in such liver diseases as von Gierke's disease, hepatoma and various other hepatocellular alterations of different origins. Since we had not accomplished the electron microscopic demonstration of G-6-Pase, although light microscopic studies on changes of the enzyme activity were done in this laboratory, this investigation was planned to obtain a satisfactory technique for ultrastructural demonstration of the enzyme activity. Unfixed frozen sections (80 micro thick) of mouse liver were washed for 2~3 minutes in a 0.4M sucrose solution (pH6.8) containing 4 mM lead nitrate and then incubated for 15~20 minutes at 32~37 degrees C in several different media to which 0.4M sucrose solution was added: A) a modification of the original Chiquoine medium, B) the first modification of the Wachstein-Meisel medium C (the second modification; the 2% lead nitrate solution was reduced in amount to 1.5 m1 instead of 3.0 ml in the medium-B). After incubation, these sections were fixed in 1% osmic acid containing sucrose, followed by embedding in Epon, ultrathin-section, mounting and staining with uranyl acetate and/or lead nitrate. By incubating the sections in the medium (B or C), satisfactory preparations were obtainable for its electron microscopic demonstration. The granular deposits of reaction products were found characteristically on the membranous component of the rough-and smooth-surfaced endoplasmic reticulum and unclear envelope. Occasional deposits were observed within cisternae or vesicles, in the nucleus, and immediate1y adjacent to the cisternal membrane and glycogen areas.
Animal ; Glucose-6-Phosphatase/metabolism* ; Liver/enzymology* ; Liver/ultrastructure ; Male ; Mice ; Microscopy, Electron

Prednisolone, a cortisol analogue, was given intraperitoneally to rats with 5, 10 or 15 doses of 5 mg. per kg. of body weight per day. Sacrificing the animals 24 hours after the last injection, unfixed frozen sections from small pieces of liver tissue were incubated and stained by a modification from the method of Wachstein and Meisel(1965) for glucose-6-phosphatase (G-6-Pase) activity. Some of the tissue blocks were processed for staining with hematoxylin and eosin for histopathological observation. Glucose-6-phosphatase activity, being represented histochemically by brownish-black deposits, was progressively increased after administrations 5 or 10 times. With 15 doses of prednisolone the general histochemical picture of G-6-Pase activity appeared to be similar to that of the control group, except for a different distribution of hepatocytes possessing strong activity. In prednisolone treated rats, the swollen hepatocytes showed a marked, cytoplasmic vacuolization and nuclear pyknosis, particularly in the periportal and midzonal areas of hepatic lobules. Some discussion of the G-6-Pase corticosteroids are presented in terms of their metabolic effects.
Animal ; Glucose-6-Phosphatase/analysis* ; Liver/drug effects ; Liver/enzymology* ; Male ; Prednisolone/pharmacology* ; Rats

Among ~20 RBC enzyme deficiencies causing hereditary hemolytic anemia (HRA), deficiencies involving three RBC enzymes such as glucose-6-phosphatase, pyruvate kinase and pyrimidine 5'-nucleodiase were known to be relatively common. The methods that have been used for RBC enzyme analysis are based on the kinetic spectrophotometry. This method, however, usually requires multiple step reactions and manual manipulations which are labor-intensive and time-consuming, and carry a greater risk of error due to their complexity. To solve this problem, we had successfully developed the multiplex enzyme analysis for galactose using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). We are now trying to adopt this method to other RBC enzymes associated with HRA. The devised method will allow simple, rapid, sensitive and reproducible quantification of RBC enzymes and should be helpful for the confirmatory diagnosis of HRA caused by RBC enzyme deficiencies.
Anemia, Hemolytic, Congenital ; Galactose ; Glucose-6-Phosphatase ; Mass Spectrometry ; Pyrimidines ; Pyruvate Kinase ; Spectrophotometry

Objective: To determine whether the adenine base editor (ABE7.10) can be used to fix harmful mutations in the human G6PC3 gene. Methods: To investigate the safety of base-edited embryos, off-target analysis by deep sequencing was used to examine the feasibility and editing efficiency of various sgRNA expression vectors. The human HEK293T mutation models and human embryos were also used to test the feasibility and editing efficiency of correction. Results: ①The G6PC3(C295T) mutant cell model was successfully created. ②In the G6PC3(C295T) mutant cell model, three distinct Re-sgRNAs were created and corrected, with base correction efficiency ranging from 8.79% to 19.56% . ③ ABE7.10 could successfully fix mutant bases in the human pathogenic embryo test; however, base editing events had also happened in other locations. ④ With the exception of one noncoding site, which had a high safety rate, deep sequencing analysis revealed that the detection of 32 probable off-target sites was <0.5% . Conclusion: This study proposes a new base correction strategy based on human pathogenic embryos; however, it also produces a certain nontarget site editing, which needs to be further analyzed on the PAM site or editor window.
Humans ; Gene Editing ; CRISPR-Cas Systems ; Adenine ; HEK293 Cells ; Mutation ; Glucose-6-Phosphatase/metabolism*

This study investigated the therapeutic effects of Inonotus obliqua extract on blood glucose, insulin, and other biochemical parameters in genetically diabetic mice (C57BL/KsJ-m+/+Lepr(db)). The mice were divided into four groups - control, Chaga 1 (dose of 0.09 mg/kg of body weight), Chaga 5 (5 times of Chaga 1), and Chaga 10 (10 times of Chaga 1)- according to supplemented dose. Inonotus obliqua extract was orally administered to the animals for 6 weeks. The body and organ (liver and kidney) weights were not different among groups. Fasting blood glucose level was significantly lower in the Chaga 5 group compared with the control (p < 0.05). Hemoglobin A1c content was significantly lower in the Chaga 5 group compared with either the control and Chaga 1 group (p < 0.05). There was no significant difference in serum insulin level among groups. The glucose-6-phosphatase activity in liver was significantly the lowest in Chaga 10 group and was significantly lower in Chaga 5 group as compared with those of control and Chaga 1 groups. Therefore, the results of this study demonstrate that Inonotus obliqua extract alleviates many of the symptoms of diabetes in genetically obese mice and may offer a possibility as a therapeutic supplement for the normalization of blood glucose levels in human with hyperglycemia and have beneficial effects in patients with noninsulin-dependent diabetes mellitus.
Animals ; Blood Glucose* ; Diabetes Mellitus, Type 2 ; Fasting ; Glucose-6-Phosphatase ; Humans ; Hyperglycemia ; Insulin ; Liver ; Mice* ; Mice, Obese ; Weights and Measures

BACKGROUND/OBJECTIVES: The goal of this study was to examine the effect of Sargassum coreanum extract (SCE) on blood glucose concentration and insulin resistance in C57BL-KsJ-db/db mice. MATERIALS/METHODS: For 6 weeks, male C57BL/KsJ-db/db mice were administrated SCE (0.5%, w/w), and rosiglitazone (0.005%, w/w). RESULTS: A supplement of the SCE for 6 weeks induced a significant reduction in blood glucose and glycosylated hemoglobin concentrations, and it improved hyperinsulinemia compared to the diabetic control db/db mice. The glucokinase activity in the hepatic glucose metabolism increased in the SCE-supplemented db/db mice, while phosphoenolpyruvate carboxykinase and glucose-6-phosphatase activities in the SCE-supplemented db/db mice were significantly lower than those in the diabetic control db/db mice. The homeostatic index of insulin resistance was lower in the SCE-supplemented db/db mice than in the diabetic control db/db mice. CONCLUSIONS: These results suggest that a supplement of the SCE lowers the blood glucose concentration by altering the hepatic glucose metabolic enzyme activities and improves insulin resistance.
Animals ; Blood Glucose ; Glucokinase ; Glucose ; Glucose-6-Phosphatase ; Hemoglobin A, Glycosylated ; Humans ; Hyperglycemia* ; Hyperinsulinism ; Insulin Resistance* ; Insulin* ; Male ; Metabolism ; Mice* ; Phosphoenolpyruvate ; Sargassum*

Glycogen storage disease is a rare metabolic disorder of significant to the anesthesiologist. The term "glycogen storage disease" is applied to a group of congenital and familial disorders characterised by depostion of abnormally large or small quantities of glycogen in the tissues. 13 types of glycogen storage diseases have been described, classified on the basis of enzyme deficiencies. Type l glycogen storage disease (von Gierke's Disease) is the most common of this constellation of syndromes. The basic defect is a deficiency of enzyme, glucose-6-phosphatase. The patient has hepatomegaly, renomegaly, stunted growth, a tend toward severe hypoglycemia and acidoais. The adverse effect of the combined anesthetic and surgical procedure during operation was reflected in a deterioration of the patients's biochemical parameters. A cardiac arrest after tonsillectomy of the patient with Von Gierke's disease was reported and this fact cmphasizes serious anesthetic problems during operation. Anesthetic management of these patients should focus on prevention of hypoglycemia and lactic acidosis. The careful frequent measuring of the acid-base status is highly recommended and is essential prior to and during andy surgical procedure. We report a case of anesthetic management for a patient with Von Gierke's desease ane review anesthetic problems for these patients.
Acidosis, Lactic ; Glucose-6-Phosphatase ; Glycogen ; Glycogen Storage Disease ; Glycogen Storage Disease Type I ; Heart Arrest ; Hepatomegaly ; Humans ; Hypoglycemia ; Tonsillectomy

Glycogen storage disease type Ia is caused by a deficiency of glucose-6-phosphatase (G6PC), which leads to glycogen accumulation in many organs including liver. We could diagnose a case of glycogen storage disease type Ia with molecular genetic analysis. A 17-year-old man visited Yeungnam university hospital because of abdominal discomfort. Clinical features were characterized by short stature, hepatosplenomegaly, accompanying hypoglycemia, hypercholesterolemia, hyperuricemia. Liver needle biopsy disclosed compatible findings of glycogen storage disease. Molecular genetic analysis of the G6PC gene was performed by direct sequencing method. We identified two mutations within the exon 5 of the G6PC gene, 727G>T and 743G>A. We report this rare case with a review of the literature.
Adolescent ; Biopsy, Needle ; Exons ; Glucose-6-Phosphatase ; Glycogen Storage Disease* ; Glycogen* ; Humans ; Hypercholesterolemia ; Hyperuricemia ; Hypoglycemia ; Liver ; Molecular Biology*