Biotin-deficient rats were raised on a purified ration containing raw egg white plus avidin. Urea synthesis and excretion were compared between the biotin-deficient and the pair-fed control rats. 24hrurinary urea excretion and the specific activities of carbamylphosphate synthetase, ornithine transcarbamylase, and arginase in the liver mitochondria fraction were no different between these two groups. The net urea production in the liver slice and in the isolated perfused liver of the biotin-deficient rats was similar to that of the pair-fed control. Thus the conclusion must be that biotin is not in urea in mea biosynthesis in the rat.
Animal ; Avitaminosis/metabolism ; Biotin* ; Liver/metabolism* ; Rats ; Urea/biosynthesis* ; MH - ; Substances: ; Urea ; Biotin

To investigate the effects of induction temperature on the expression product and the impact of urea concentration on the purification, HIV-1 Gag inclusion bodies from E. coli induced at 30 degrees C (IB30) and 37 degrees C (IB37) were dissolved with urea of different concentrations. The solubility and yield of refolding were compared. IB30 were dissolved with 2 mol/L and 8 mol/L urea, and then purified with chromatography. IB30 were found easier to be solubilized in low concentration of urea and easier to be refolded than IB37. Furthermore, compared to the IB30 dissolved in 8 mol/L urea, Gag protein solubilized in 2 mol/L urea was purified to higher purity with gel filtration (GF) and ion exchange (IEX) chromatography. Gag inclusion body induced at lower temperature may contain more protein with native-like or reversibly-denatured structures, and solubilization in the presence of low concentrations of urea can help to retain these structures. This study has provided new insights into the purification of proteins from inclusion bodies.
Escherichia coli ; genetics ; metabolism ; HIV-1 ; genetics ; Humans ; Inclusion Bodies ; metabolism ; Protein Denaturation ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Temperature ; Urea ; chemistry ; gag Gene Products, Human Immunodeficiency Virus ; biosynthesis ; genetics

Relative deficiency in production of glycoprotein hormone erythropoietin (Epo) is a major cause of renal anemia. This study planned to investigate whether the hypoxia-regulated system of Epo expression, constructed by fusing Epo gene to the chimeric phosphoglycerate kinase (PGK) hypoxia response elements (HRE) in combination with cytomegalovirus immediate-early (CMV IE) basal gene promoter and delivered by plasmid intramuscular injection, might provide a long-term physiologically regulated Epo secretion expression to correct the anemia in adenine-induced uremic rats. Plasmid vectors (pHRE-Epo) were synthesized by fusing human Epo cDNA to the HRE/CMV promoter. Hypoxia-inducible activity of this promoter was evaluated first in vitro and then in vivo in healthy and uremic rats (n = 30 per group). The vectors (pCMV-Epo) in which Epo expression was directed by a constitutive CMV gene promoter served as control. ANOVA and Student's t-test were used to analyze between-group differences. A high-level expression of Epo was induced by hypoxia in vitro and in vivo. Though both pHRE-Epo and pCMV-Epo corrected anemia, the hematocrit of the pCMV-Epo-treated rats exceeded the normal (P < 0.05), but that of the pHRE-Epo-treated rats didn't. Hypoxia-regulated system of Epo gene expression constructed by fusing Epo to the HRE/CMV promoter and delivered by plasmid intramuscular injection may provide a long-term and stable Epo expression and secretion in vivo to correct the anemia in adenine-induced uremic rats.
Anemia/blood/*therapy ; Animals ; Base Sequence ; Blood Urea Nitrogen ; Cell Hypoxia ; Creatinine/blood ; Erythropoietin/biosynthesis/*genetics/secretion ; Gene Expression Regulation ; Genes, Reporter ; Genetic Therapy ; HeLa Cells ; Humans ; Injections, Intramuscular ; Kidney/pathology ; Luciferases, Firefly/biosynthesis/genetics ; Molecular Sequence Data ; Plasmids/*genetics ; Promoter Regions, Genetic ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins/biosynthesis/genetics/secretion ; Response Elements ; Transcriptional Activation ; Uremia/blood/*therapy

INTRODUCTIONSeizure is the most common paediatric neurological disease which occurs in ten percent of children. In approaching a convulsive patient, finding the causes of seizure is essential, and the patient's history as well as the physical examination are important. The role of routine laboratory tests for children's seizures (except neonates) is undetermined, but checking for serum sodium, glucose, calcium and urea routinely has been advised. The purpose of this study was to determine the diagnostic efficacy of these serum chemistry tests in the seizures of children older than one month of age.

METHODSIn this descriptive, retrospective study, medical records of 302 hospitalised children with seizure were reviewed. Results of laboratory tests, like sodium, calcium, blood glucose and urea levels, pertinent history and physical examination, and the change in patient management based on serum chemistry test results, were analysed. All the children in the study were classified as having seizure with or without fever.

RESULTSIn 302 hospitalised children with seizure, about ten percent of 938 tests were abnormal. 27.7 percent of these abnormal results were seen in 1-12-month-old infants. Only 11 percent of abnormal tests (1.3 percent of total tests) might have caused a seizure. Also, 0.2 percent of the results could not be predicted from the history or physical examination, which was conducted in patients younger than one year of age.

CONCLUSIONRoutine determination of serum chemistry values in seizures of children does not contribute to therapy, and are costly and time-consuming. It may not be helpful and informative unless the patient is less than one year of age.

Blood Chemical Analysis ; methods ; Calcium ; blood ; Chemistry, Clinical ; methods ; Child ; Child, Preschool ; Female ; Glucose ; biosynthesis ; Humans ; Infant ; Male ; Retrospective Studies ; Seizures ; blood ; diagnosis ; Sodium ; blood ; Treatment Outcome ; Urea ; blood

OBJECTIVETo investigate the effects of Shenkang pill on renal function and extracellular matrix secretion on the diabetic rats.

METHODThe diabetic rat models were induced by intraperitoneal injection of streptozotocin (STZ) and randomly divided into 3 groups' model control group; Capoten group and Shenkangwan group. Some normal other rats were used as normal control group. All rats were treated with corresponding drugs for 8 weeks. During and after the treatment, the general state, blood and urine glucose levels, excretion rate of the 24 hour urine protein and albumin, serum creatinine and blood urea nitrogen contents, kidney weight and relative kidney weight were measured. The mRNA of fibronectin(FN) in the kidney also detected by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR).

RESULTDiabetes mellitus and renal lesions occurred in the three model groups. The expression of FN mRNA of the kidney in diabetic rats increased obviously. Shenkang pill could improve the general state and renal function of the diabetic rats, decrease the blood glucose levels and the excretion rate of the 24 hour urine protein and albumin, reduce the expression of FN mRNA in kidney.

CONCLUSIONShenkang pill has a certain protective effect on the diabetic kidney.

Animals ; Blood Glucose ; metabolism ; Blood Urea Nitrogen ; Diabetic Nephropathies ; chemically induced ; metabolism ; pathology ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Fibronectins ; biosynthesis ; genetics ; Glycated Hemoglobin A ; metabolism ; Kidney ; metabolism ; Male ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Wistar ; Streptozocin

OBJECTIVETo investigate the effects of Haikun Shenxi on the expression of platelet-derived growth factor-BB (PDGF-BB) and mRNA in renal tissue of rats with adriamycin nephropathy.

METHODRat model was established by unilateral nephrectomy and injecting adriamycin intraperitoneally. The adriamycin-induced nephrotic rats were randomly divided into 6 groups: normal group, sham operation group, model group, lotensin treatment group, Haikun Shenxi low and high dose treatment groups (0.77, 0.08 mg x kg(-1). Ten weeks later, the 24 hour urine protein and blood biochemistry examinations and renal pathologic changes were observed, and the expression of PDGF-BB and mRNA was measured using immunohistochemical method.

RESULTCompared with model group, proteinuria and the levels of serum creatinine (Scr) , urea nitrogen (BUN) were decreased obviously in both Haikun Shenxi low and high dose groups. The expression of PDGF-BB and mRNA was mostly presented in cytoplasm of renal tubular epithelial cells and mesangial area, and it could be reduced significantly after treatment (P < 0. 05).

CONCLUSIONThe level of PDGF-BB and mRNA is high in renal tissue of adriamycin-induced nephrotic rats. This progress could be effectively inhibited by Haikun Shenxi and the mechanism may be that it can control the excessive expression of PDGF-BB and mRNA.

Animals ; Blood Urea Nitrogen ; Creatinine ; blood ; Doxorubicin ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Gene Expression Regulation ; drug effects ; Glomerular Mesangium ; drug effects ; metabolism ; pathology ; Glomerulosclerosis, Focal Segmental ; chemically induced ; genetics ; metabolism ; Immunohistochemistry ; In Situ Hybridization ; Kidney ; drug effects ; metabolism ; pathology ; Male ; Medicine, Chinese Traditional ; Phaeophyta ; chemistry ; Platelet-Derived Growth Factor ; biosynthesis ; genetics ; Polysaccharides ; chemistry ; isolation & purification ; pharmacology ; Proto-Oncogene Proteins c-sis ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Wistar

The effect of NaCl plus 3% chitosan on the systolic blood pressure of spontaneously hypertensive rats (SHR) were evaluated and compared with NaCl plus KCl (NaCl, 49.36% + KCl 49.36%) and chitosan or NaCl treatment alone. In SHR, administration of NaCl plus chitosan (44 mM Na/day) for two months significantly decreased the systolic blood pressure greater than of NaCl plus KCl and NaCl alone. NaCl plus chitosan resulted, though not statistically significant, in decreased urinary Na+ excretion and decreased blood urea nitrogen levels. Urinary creatinine of NaCl plus chitosan was slightly decreased compared to 3 treated groups. Serum electrolytes levels, however, remained unchanged. The combination of NaCl and chitosan may be superior to the conventional use of NaCl plus KCl or NaCl alone in the prevention of hypertension. Even though these supplementary diets have demonstrated potential anti-hypertensive effects in the experimental animal model, further research is needed before any recommendations can be made.
Angiotensin I/blood ; Angiotensin II/biosynthesis ; Animals ; Blood Pressure/*drug effects/physiology ; Blood Urea Nitrogen ; Body Weight/drug effects ; Chitosan/*administration & dosage ; Chlorides/blood/urine ; Creatinine/urine ; Heart/physiology ; Histocytochemistry ; Hypertension/*prevention & control ; Kidney/physiology ; Male ; Potassium/blood/urine ; Potassium Chloride/administration & dosage ; Random Allocation ; Rats ; Rats, Inbred SHR ; Sodium/blood/urine ; Sodium Chloride, Dietary/*administration & dosage ; Systole/drug effects/physiology