MTHFR gene polymorphism as a risk factor for diabetic nephropathy and macrovascular disease in NIDDM patient / 대한내과학회지

BACKGROUND: Diabetic nephropathy develops in 20-30% of patients with non-insulin dependent diabetes mellitus (NIDDM). Poor glycemic control, hypertension and duration of diabetes are known as risk factors for the development of diabetic nephropathy and there is high prevalence of diabetic nephropathy in the patients who have familial history of diabetic nephropathy, so it has been assumed that genetic factor is associated with the background of its occurences. Recently it has been observed that a cytosine to thymidine substitution of the methylenetetrahydrofolate reductase (MTHFR) gene at nucleotide 677 (C677T) was related to diabetic nephropathy in patients with NIDDM and MTHFR gene polymorphism was also known to predispose to vascular disease. This study was performed to investigate whether MTHFR gene polymorphism is associated with the development of diabetic nephropathy and macrovascular disease in NIDDM patients. METHODS: The study population consisted of 243 NIDDM patients (duration> OR = 10 years). Nephropathy was defined by 24 hour urinary protein excretion of more than 500 mg. The MTHFR gene fragment was extracted using the polymerase chain reaction. The presence of the mutation was identified by HinfI digestion, which cuts at the mutation site, followed by 2.5% metaphore agarose electrophoresis and ethidium bromide staining. Statistical differences in genotype distribution and allele frequencies among the groups were assessed by the chi-square test. RESULTS: There was no difference in clinical characteristics except the prevalence of hypertension and diabetic retinopathy between nephropathy group and non-nephropathy group. The data do not show any difference of genotype distribution or allele frequencies between patients with or without diabetic nephropathy and macrovascular disease CONCLUSION: With the above results, it is assumed that there are no significant relationships among MTHFR gene polymorphism, diabetic nephropathy, and macrovascular disease.

Rapid Molecular Detection of Escherichia coli O157:H7 / 감염

BACKGROUND: Sorbitol fermenting Escherichia coli O157 were reported. And E. coli O157:H7 produce various Shiga toxin (Stx) such as Stx1, Stx2, or variants of Stx2. In this study, we tried to establish laboratory methods that detect E. coli O157:H7 quickly and precisely by analyzing sensitivity of colony hybridization test and PCR technique. METHODS: Stx1-producing E. coli ATCC 43890, Stx2-producing E. coli ATCC 43889, and Stx2vha- producing E. coli ATCC 51435 were tested. Three strains of E. coli were diluted with 0.1 g of diarrheal stools from 107 CFU to 101 CFU respectively. The stool samples were incubated overnight in MacConkey agar plates. A mean of 63 colonies were hybridized by stx1- and stx2-specific oligonucleotide probes. PCR for stx1 gene and stx2 gene was done after overnight- incubation of stool samples in the LB broth with vancomycin (6 ug/mL). Positive colonies by colony hybridization were confirmed by PCR for stx1 gene and stx2 gene. RESULTS: Colony hybridization test could detect Stx1-producing E. coli at 103 CFU per 0.1 g of stool, Stx2-producing E. coli at 105 CFU per 0.1 g of stool, and Stx2vha-producing E. coli at 104 CFU per 0.1 g of stool. PCR technique after enrichment in LB broth with vancomycin (6 ug/mL) could detect stx1-, stx2-, and stx2vha-containing E. coli at 10 CFU per 0.1 g of stool respectively. CONCLUSOIN: A combination of colony hybridization and PCR after enrichment in broth with vancomycin (6 ug/mL) is useful for the rapid and precise diagnosis of infections of Shiga toxin-producing E. coli O157:H7.

Does Albumin Infusion Enhance the Diuretic Action of Furosemide in NePhrotic Syndrome? / 대한신장학회잡지

There have been controversies on the effect of albumin in treating edema in nephrotic syndrome patients. We evaluated the additive diuretic effect of coadministration of furosernide with albumin in the six patients with nephrotic syndrome. We administered 160mg of furosemide intravenously for 1 hour with 100rnl of 20% albumin or 5% dextrose by random cross-over design. The urine and plasma furosemide concentrations were measured by HPLC. After the administration of furosemide alone, urine volume, urinary excretions of sodium and chloride were increased significantly compared to those of basal state (P<0.05). But, coadministration of furose-mide with albumin did not increase significantly the urine voume (2285+/-445ml vs. 3023+/-715ml), urinary excretions of sodium (194+/-58rnmol/day vs. 282+/-85 mmol/day) and chloride (213+/- 54mmoVday vs. 286+/- 74mmoVday) comparing to those of furosemide only cases. Addition of albumin to furosemide did not significantly changed pharmacokinetic parameters such as AUC (28.3+/-5.5ug/ml hr vs 36.0+/-6.7ug/ml hr), total plasma clearance (115+/-30mVmin vs 108+/-41ml/min), volume of distribution (0.13+/-0.02L/kg vs 0.10+/- 0.01L/kg), elirnination half life (1.4+/-0.3hr vs 1.5+/-0.3hr), and urine furosemide excretion (44+/-8% vs 43+ 10%). We concluded that albumin infusion did not enhance the diuretic action of furosemide pharmacodynamically and pharmacokinetically in patients with nephrotic syndrome.

Imaging of Hyperparathyroidism / 대한내분비학회지

No abstract available.

Changes in White Blood Cells after Transplantation of Vascular Smooth Muscle Cells Retrovirally Transduced with Granulocyte Colony-Stimulating Factor Gene in Rats / 대한혈액학회지

BACKGROUND: The granulocyte colony-stimulating factor (G-CSF) is crucial in neutrophil regulation. Since recombinant G-CSF became clinically available, it has been widely used in the treatment of neutropenia. Ex vivo therapy of recombinant G-CSF, however, requires large dose and frequent administration, which brings financial burden on the patients. To overcome disadvantages of ex vivo therapy, we have tried to make an in vivo G-CSF delivery system in rat using gene therapy technique. METHODS: We have tried to make an in vivo G-CSF delivery system using transduced vascular smooth muscle cells with G-CSF gene in a rat model. Retroviral vector plasmid containing rat G-CSF gene was made employing LXSN and LNFX plasmid. Recombinant retrovirus was produced from PA317 packaging cells. Infection of the vascular smooth muscle cells with the virus and selection with G418 was done in vitro. These transduced cells were transplanted to the balloon-injured carotid arteries of Fisher 344 rats, and complete blood count as well as differentials were measured in sequence. RESULTS: The virus titer was three times greater in case of LNFG than LGSN, whereas G-CSF production from infected vascular smooth muscle cell was lower in LNFG vector (0.1ng/106cells/day) than in LGSN vector (0.4ng/106cells/day). The increment of WBC count was observed until 25 days after transplantation, being 9,600 +/- 1,000/uL on seventh day after transplantation, which was significantly higher than that of controls, 7,300 +/- 540/uL. The levels of neutrophil increased gradually after transplantation, reached to the peak after 1 week (3,250 +/- 1,099/uL in case of neutrophil count and 30 +/- 10% in case of differentials). The duration of increment, however, was relatively short, neutrophil count being decreased to the basal level within 4 weeks. CONCLUSION: The effective increase of neutrophil count with low dose of G-CSF produced from vascular smooth muscle cells could make this gene therapy feasible in the clinical settings only if the problem of short duration of effect could be solved.

Gene Therapy / 대한내분비학회지

No abstract available.

Retrovirus-Mediated Herpes Simplex Virus Thymidine Kinase Gene Therapy for the Prevention of Stenosis in Rat Carotid Artery Injury Model

BACKGROUND: Herpes simplex virus thymidine kinase (HSVtk) phosphorylates the prodrug ganciclovir to a nucleoside analog that inhibits DNA synthesis, causing cell death. Neighbouring nontransfected cells may be affected through a 'bystander effect', thereby amplifying the antiproliferative actions. This study was carried out to determine whether retrovirus-mediated HSVtk gene therapy could reduce intimal hyperplasia and prevent stenosis following balloon injury of the rat carotid artery. METHODS: A replication-defective recombinant retroviral vector containing HSVtk cDNA (LtkSN) was constructed. Cultured primary rat smooth muscle cells (SMCs) infected with this vector (SMC/LtkSN) were transplanted to the balloon injured rat right carotid artery. One week after transplantation, HSVtk gene therapy group was administered a 2-week treatment of ganciclovir (30 mg/kg/d). Three weeks after balloon injury and SMC/LtkSN transplantation, carotid arteriography was performed and carotid arteries were perfusion-fixed for histologic examination. RESULTS: Carotid arteriographic evaluation comparing with the uninjured left carotid artery showed that the mean luminal diameter of HSVtk gene therapy group (n=5, 85+/-3%) was significantly larger than that of balloon injury only group (n=5, 65+/-5%). The neointimal mass of HSVtk gene therapy group was less than that of balloon injury only group. SMC/LtkSN transplantation without ganciclovir treatment group (n=3) showed asymmetric intimal proliferation probably because of gravitational pooling of seeding. There were inflammatory cell infiltrations at the gravity dependent portion of HSVtk gene therapy group. CONCLUSION: Retrovirus-mediated HSVtk gene therapy following balloon injury of the rat carotid artery reduced neointimal expansion and arteriographic stenosis.

Gene Therapy Using Retroviral Vector Containing Rat Erythropoietin / 대한혈액학회지

BACKGROUND: Erythropoietin (EPO) is crucial to red blood cell regulation. Since the recombinant EPO was clinically available, it has been widely used in the treatment of anemia associated with disorders such as chronic renal failure, cancer and acquired immunodeficiency syndrome. Ex vivo therapy of recombinant EPO, however, requires large dose and frequent administration, which gives a financial impact to the patients. To make in vivo delivery system of EPO will be valuable to the patients who need EPO for a long time. METHODS: We have tried to make an in vivo EPO delivery system using transduced vascular smooth muscle cells with EPO gene in a rat model. Recombinant retroviral vector containing EPO gene was made employing LXSN and LNFX plasmid. Recombinant retrovirus was produced from PA317 packaging cell. Infection of the vascular smooth muscle cells with the virus and selection with G418 was done in vitro. These transduced cells were transplanted to the balloon-injured carotid artery, and hemoglobin and hematocrit as well as reticulocyte were measured in sequence. RESULTS: The virus titer was ten times greater in case of LNFEp than LEpSN, whereas EPO production from infected vascular smooth muscle cell was similar between LEpSN and LNFEp vectors (67mU/106cell/day and 68mU/106cell/day, respectively). The levels of hemoglobin and hematocrit increased gradually after transplantation of the transduced cells with LEpSN, reached to peak after 3 weeks (18.4+/-0.63gm/dL in case of hemoglobin and 50.7+/-0.62% in case of hematocrit) and remained thereafter. The percentage of reticulocyte was elevated from the 4th day to the 14th day after transplantation and returned to normal. CONCLUSION: The low dose required to the elevation of RBC mass and long sustained effect of transduced smooth muscle cell could make this gene therapy feasible to the clinical conditions.

A Case of DiGeorge Syndrome Associated with Complex Cardiovascular Anomalies / 대한내과학회지

DiGeorge syndrome is the developmental anomalies of the third and fourth pharngeal pouches. Recently, damages or abnormal development of the neural crest is suggested as a possible pathogenetic factor, because neural crest cells play a crucial role in development of pharyngeal pouch derivatives, e.g. thymus and parathyroid glands, as well as the aortic arches and conotruncal part of the heat. Most cases have abnormal findings of chromosome 22 and are sporadic, but familial cases have been described. Typical features of DiGeorge syndrome are congenital heart disease, aplasia or hypoplasia of the thymus and parathyroid glands and facial dysmorphism. The main problems and cause of death are severe congestive heart failure due to cardiac anomlies, hypocalcemic complications or immunocompromised conditions. As these results, most cases were expired at infantal period or early childhood. Recently, we have a case of Digeorge syndrome which was associated with complex cardiovascular anomalies(tetralogy of Fallot, atrial septal defect, right aortic arch, left hemitruncus), severe hypocalcemia, aplasia of thymus and facial dysmorphism.

Effect of Antiliyolytic Agents on Glueose Metabolism in Thyrotoxic Patients / 대한내분비학회지

Decreased glucose tolerance is often found in patients with thyrotoxicosis but the pathogenetic mechanisms are poorly understood. Since the concentrations of free fatty acid are usually elevated due to increased lipolysis in thyrotoxicosis, the preferential oxidation of the free fatty acids may explain the decreased glucose tolerance in hyperthyroidism. The aim of this study was to investigate whether lowering plasma free fatty acid(FFA) by acipimox, a long-acting antilipolytic agent, could affect glucose metabolism in thyrotoxicosis. We performed intravenous glucose tolerance test with acipimox or placebo in 6 untreated thyrotoxicmen and 6 age-and body mass index(BMI)-matched controls. The following results were obtained.1) The basal plasma FFA concentration in thyrotoxic patients were significantly higher than those in controls(997.0+-303.4 uEq/L vs. 290.5+-169.1 uEq/L; p<0.01). 2) Plasma FFA concentrations decreased rapidly with acipimox ingestion in both controls and thyrotoxic patients.3) Plasma glucose concentrations were significantly lower with acipimox ingestion than with placebo in thyrotoxic patients from 17min after intravenous glucose load and to the end of the study.4) Plasma insulin concentrations in thyrotoxic patients with acipimox ingestion were higher at 5, 7 min after iv glucose load.5) In thyrotoxic patients, glucose disappearance rate(K_glucose) in acipimox treatment was significantly higher than that in placebo treatment(2.44+-0.84 vs. 1.58+-0.37;p<0.05). 6) K_glucose values were inversely correlated with basal FFA concentrations(r=-0.58, p<0.05). In summary, in thyrotoxic patients with elevated plasma FFA levels, acipimox lowered plasma FFA, which in turn improved glucose tolerance.

Insulin Resistance, Body Fat Distribution, and Sex Hormones in Healthy Men and Premenopausal Women / 대한내분비학회지

It is well known that obesity central obesity is associated with insulin resistance and some studies reported that sex hormones were associated with insulin resistance. Recently, low levels of sex-hormone binding globulin(SHBG), an indirect index of androgenicity, have been observed to predict the development of non-insulin dependent diabetes mellitus(NIDDM) in women and SHBG has been proposed as a marker for insulin resistance. In contrast to findings in women, decreased SHBG did not predict the occurrence of NIDDM in men, so it is suggested that sex hormones may have a different role for insulin resistance between men and women. To investigate the difference of the associations among the body fat distribution, sex hormone and insulin sensitivity index in men and women, we measured body-mass index(BMI) and waist to hip circumference ratio(WHR) and concentrations serum SHBG, total testosterone, free testosterone, and dehydroepiandrosterone sulfate(DHEA-S) concentrations in 29 healthy adults(men:19, women:10) who showed normal glucose tolerance. Insulin sensitivity index(M/I) was measured by euglycemic hyperinsulinemic clamp. There were no differences in age, BMI, fasting plasma glucose, insulin and free fatty acid levels between men and women. WHR of men is higher than that of women(0.82+-0.01 vs. 0.73+-0.01, p=0.002). Insulin sensitivity index(M/I) is similar in men and women(7.80+-0.71 mg/kg/min/uU/ml X 100 vs. 9.74+-0.89 mg/kg/min/uU/ml X 100, p=0.196).In Pearson's correlation, M/I was significantly correlated with BMI(r=-0.69, p<0.01) and WHR(r=-0.68, p<0.01) in men and DHEA-S(r=-0.68, p<0.05) and SHBG(r=0.61, p=0.056) concentrations in women.In multiple regression analysis, M/I had the most significant association with BMI(R

Cell density dependence of growth characteristics of rat thyroid cells(FRTL-5) stimulated by TSH and IGF-I / 대한내분비학회지

No abstract available.

A case of functioning adrenal rest tumor of liver / 대한내과학회지

No abstract available.

Combined estrogen-progesterone(Divina@) treatment in postmenopausal women with special reference to serum lipoprotein patterns / 대한내분비학회지

No abstract available.