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Volume 24(3); June 2000
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Original Articles
Platelet Aggregability in Type 2 Diabetics.
Chang Hun Lee, Nam Il Cheon, Yeon Sang Lee, Dong Hyeok Cho, Hyun Ho Shin, Jung Min Kim, Dae Ho Lee, Dong Jin Chung, Min Young Chung, Tai Hee Lee
Korean Diabetes J. 2000;24(3):300-309.   Published online January 1, 2001
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AbstractAbstract PDF
BACKGROUND
Increased platelet aggregability is known to be one of the important risk factors for diabetic vascular complications. The relationship between platelet aggregability and diabetic macrovascular complications, varying severity of diabetic microvascular complications and other possible factors was evaluated in Korean adults. METHODS: Platelet aggregability was measured with platelet rich plasma by addition of adenosine diphosphate (ADP) in 45 cases. Normal control group (n=15) was compared with diabetics without macrovascular complications (n=15), diabetics with macrovascular complications (n=15) and several groups divided accoring to the severity of microvascular complications. RESULTS: 1) The mean maximum value of platelet aggregation was 70.3+/-5.3% in control group, and 80.0+/-7.3% in diabetics (p<0.005). 2) The mean maximum value of platelet aggregation was 78.0+/-5.5% in diabetics without macrovascular complications and 83.5+/-7.1% in diabetics with macrovascular complications (p=0.093). 3) The mean maximum value of platelet aggregation was 77.0+/-5.1% in normoproteinuria group, 78.1+/-7.3% in microproteinuria group, and 82.9+/-6.2% in overt proteinuria group (p=0.083). 4) The mean maximum value of platelet aggregation was 77.2+/-6.8% in diabetes without neuropathy group and 82.9+/-6.2% in diabetes with neuropathy group (p=0.114). 5) The mean maximum value of platelet aggregation was 79.3+/-4.9% in diabetes with normal funduscopic findings, 80.2+/-7.3% in diabetes with background retinopathy and 81.6+/-7.9% in diabetes with proliferative retinopathy (p=0.852). 6) Blood glucose showed positive correlations with the mean maximum platelet aggregation ( =0.529, p<0.005). CONCLUSION: The elevated mean maximum value of platelet aggregation was found in diabetics and there were no significant differences between macrovascular complications and between varying severity of retinopathy, neuropathy and proteinuria. Blood glucose showed positive correlations with mean maximum platelet aggregation. Hyperglycemia was a major risk factor affecting platelet aggregation in diabetics and its control may play an important role in prevention of diabetic vascular complications.
In Vitro Expansion and Differentiation of Islet Precursor Cells from Cultured Neonatal Porcine Pancreatic Tissue.
Yu Bae Ahn, Kun Ho Yoon, Sun Hee Seo, Seung Hyun Ko, Ki Ho Song, Je Ho Han, Soon Jip Yoo, Hyun Sik Son, Moo Il Kang, Bong Yun Cha, Kwang Woo Lee, Ho Young Son, Sung Koo Kang
Korean Diabetes J. 2000;24(3):310-322.   Published online January 1, 2001
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AbstractAbstract PDF
BACKGROUND
Neonatal porcine pancreas is an attractive alternative source for islet transplantation because of its growth potential and availability. Porcine neonatal pancreatic cell clusters (NPCCs) consist mainly of protodifferentiated cells expressing both the duct cell marker pancytokeratin and islet hormones. In this study, we investigated to expand and mature the pancreas duct cells contained in porcine NPCCs with extracellular matrix. METHODS: For NPCCs, pancreas obtained from neonatal pigs were minced, digested with collagenase and cultured overnight. Then NPCCs were further dispersed to small cell groups and cultured on HTB-9 extracellular matrix: the tissue attached and formed monolayer patches. At the 3rd and 8th days, tissue was fixed, immunostained for pancytokeratin (panCK), vimentin (VT) and islet hormones. RESULTS: During 5 days culture, the total cell numbers increased 3.2 fold on the matrix, and 1.6 fold on the sticky dish, respectively. Insulin positive cells (Ins+) were 6.0% of total cells at day 3 and increased 1.6 fold in numbers at day 8. There was significant increase in DNA content of NPCCs in monolayers on both sticky dishes and HTB-9 matrix. In contrast, insulin content of both groups decreased during culture periods. Until 8 days of culture after dispersion of porcine NPCC, most duct cells costained with panCK and VT. CONCLUSION: We observed NPCCs were composed of many of duct cells which were known to be endocrine precursor cells and monolayer culture of NPCC withextracellular matrix resulted in the proliferation and differentiation of pancreatic duct cells.
Expression of Gal alpha1,3 Gal Antigen and Galactosyl Transferase mRNA in Porcine Neonatal Pancreatic Tissue.
Kyong Soo Park, Yoon Young Kim, Jeong Mi Kim, Yu Bae Ahn, Kun Ho Yoon, Bong Yun Cha, Ho Young Son, Hong Kyu Lee
Korean Diabetes J. 2000;24(3):323-330.   Published online January 1, 2001
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AbstractAbstract PDF
BACKGROUND
Neonatal porcine pancreatic tissue may be a potential source of islet transplantation in patients with type 1 diabetes. Gal 1,3 Gal antigen (Gal epitope) is a xenoantigen which is responsible for hyperacute xenograft rejection. The aim of this study is to evaluate the expression of Gal epitope and galactosyl transferase mRNA in porcine neonatal pancreatic tissue. METHOD: Porcine neonatal pancreatic cell clusters (NPCCs) were isolated using collagenase and incubated in various culture condition. They were stained with Gal specific lectin for the detection of Gal epitope. Expression of 1,3 galactosyl transferase mRNA was assessed by semiquantitative RT-PCR. RESULTS: Gal epitope was expressed in both neonatal porcine pancreas and cell clusters. Most of Gal epitope expressed cells were endothelial cells and ductal epithelial cells. A small number of cells stained positive for insulin were also positive for Gal epitope. In some area of monolayer culture of porcine neonatal islet cluster, scattered insulin positive cells coexpressed the Gal epitope. The expression of 1,3 galactosyl transferase mRNA were lower in islet than other tissues. Culture using extracelluar matrix or 3D gel increased the expression of 1,3 galactosyl transferase mRNA levels. CONCLUSION: Gal epitope was expressed in ductal epithelial cells and some of beta cells of porcine neonatal pancreatic tissue. Expression of Gal epitope in porcine neonatal pancreatic cell cluster may be a problem that needs to be solved before porcine NPCCs can be used in human.
Metabolic Phenotype of Glycogen Synthase Gene Inhibition in Human Skeletal Muscle Cells.
Jae Joon Koh, Kyong Soo Park, Jeong Mi Kim, Seong Yeon Kim, Hong Kyu Lee, Theodore P Ciaraldi, Robert R Henry
Korean Diabetes J. 2000;24(3):331-339.   Published online January 1, 2001
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AbstractAbstract PDF
BACKGROUND
Glycogen synthase (GS) is the rate-limiting enzyme controlling non-oxidative glucose disposal in skeletal muscle. Reduction in GS activity and impaired insulin responsiveness are characteristic features of skeletal muscle in type 2 diabetes that contribute to glucose intolerance. These properties also exist in human skeletal muscle cell cultures from type 2 diabetic subjects. The aim of study is to determine the effect of an isolated reduction in GS on glucose metabolism and if this change can generate a diabetes-like state. METHODS: Cultured skeletal muscle cells from non-diabetic subjects were treated with antisense oligodeoxynucleotides (ODN) to GS to interfere with expression of the gene for 6 days. GS activity, protein expression, glycogen synthesis and cellular glycogen content were measured. RESULTS: Treatment with antisense ODN reduced GS protein expression by 70% compared to control (scrambled) ODN (p<0.01). Both total GS activity and that measured at 0.1 mM G-6-P were reduced by antisense ODN treatment. Insulin responsiveness of GS was also halved. Basal GS FV0.1 was decreased in both antisense ODN and control ODN treated cells and antisense treated cells did not show increase in GS FV0.1 in response to insulin stimulation. Glucose incorporation into glycogen under basal conditions was unaltered after antisense ODN treatment, though no further stimulation in response to insulin was observed. Yet both cellular glycogen content and glycogen synthesis were lower in antisense ODN treated cells compared to control ODN treated cells. CONCLUSIONS: Reduction in GS expression in human skeletal muscle cell impair GS activity and insulin responsiveness but does not replicate the abnormalities of glycogen synthesis found in cultured diabetic skeletal muscle cells.
The Effect of BCG Vaccine on Recent Onset Type 1 Diabetes Mellitus Patients.
Jeong Heon Oh, Sei Hyun Baik, Kyung Mook Choi, Nan Hee Kim, Ie Byung Park, Dong Seop Choi
Korean Diabetes J. 2000;24(3):340-347.   Published online January 1, 2001
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AbstractAbstract PDF
BACKGROUND
Type 1 diabetes mellitus (Type 1 DM) results from autoimmune destruction of -cells of the pancreas. Many treatments aimed at inducing remission of newly diagnosed type 1 DM or preventing of type 1 DM in high risk group are being conducted. BCG is known to modulate the development of spontaneous diabetes in animal model of type 1 DM. In some studies, single injection of BCG induced clinical remission in recent onset type 1 DM patients. However, the effect of BCG on human is still controversial. Thus, we performed a prospective study to evaluate the effect of BCG on type 1 DM. METHODS: We enrolled a total of 23 type 1 DM patients within 6 months period. Randomly selected 14 patients were injected 0.1 ml BCG intradermally and 9 patients were injected normal saline. Fasting and postprandial 2 hour C-peptides, and insulin requirements were measured in all patients at enrollment and at 6, 12 and 24 months after BCG vaccination. RESULTS: At enrollment, there was no significant difference in age, sex, duration of diabetes, HbA1-C, body mass index, fasting and postprandial 2 hour C-peptides, and insulin requirement between BCG group and control group. During follow-up, there was no significant difference in fasting and postprandial 2 hour C-peptides. However postprandial 2 hour C-peptides in BCG group were higher than those in control group at 12 and 24 months (p-value>0.05). Insulin requirements also were lower in BCG group than in control group at 12 and 24 months (p-value>0.05). Clinical remission has been sustained in 2 BCG vaccinated patients at 6 and 12 months. In one of the two patients, remission was sustained for 36 months. CONCLUSION: BCG vaccine is safe and convenient to use, however, a large study is warranted for the use of BCG as a therapy of type 1 DM.
The Effects of Troglitazone on Vascular Smooth Muscle Cell Proliferation.
Yun Jae Chung, Kyeong Min Min, Eun Young Oh, Jae Hoon Chung, Yong Ki Min, Myung Shik Lee, Moon Kyu Lee, Kwang Won Kim
Korean Diabetes J. 2000;24(3):348-355.   Published online January 1, 2001
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AbstractAbstract PDF
BACKGROUND
Elevated fasting and postprandial insulin levels are frequently observed in patients with obesity and hypertension as well as type 2 diabetes mellitus. This phenomenon has been suggested as an independent risk factors for atherosclerotic cardiovascular diseases. Troglitazone, an insulin-sensitizing antidiabetic agent, has been shown to inhibit atherosclerotic process, but its mechanism of action is not yet elucidated. This study was undertaken to examine the effects of troglitazone, a peroxisome proliferator- activated receptor- (PPAR ) ligand, on vascular smooth muscle cell proliferation. METHODS: Aortic smooth muscle cells were isolated from Sprague-Dawley rats and the effects of several different agonists (insulin, ET-I, IGF-I) on cellular DNA synthesis were measured and compared with the effects of troglitazone. In addition, the mRNA of PPARgamma gene in rat aortic smooth muscle cells(RASMCs) was detected by RT-PCR methods. RESULTS:1. Insulin, endothelin-I and IGF-I significantly stimulated DNA synthesis in RASMCs (p<0.05). 2. Insulin-induced DNA synthesis was not significantly inhibited by coincubation with wortmannin or LY294002 but inhibited by PD98059. 3. Troglitazone significantly inhibited insulin, endothelin-I and IGF-I-induced DNA synthesis in RASMCs (p<0.05, respectively). 4. PPAR mRNA was detected in RASMCs by RT-PCR and its expression did not significantly increase by troglitazone treatment. CONCLUSION: Troglitazone could inhibit agonist-induced proliferation of vascular smooth muscle cells and might be a useful agent for treatment as well as prevention of atherosclerosis.
Insulin Secretion and Insulin Sensitivity in Korean Subjects with Impaired Glucose Intolerance.
Dong Jun Kim, Jong Ryul Hahm, In Kyoung Jeong, Tae Young Yang, Eun Young Oh, Yoon Ho Choi, Jae Hoon Chung, Yong Ki Min, Myung Shik Lee, Moon Kyu Lee, Kwang Won Kim
Korean Diabetes J. 2000;24(3):356-364.   Published online January 1, 2001
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AbstractAbstract PDF
BACKGROUND
Although insulin resistance has been known to be a primary defect causing type 2 diabetes in Pima Indians and Caucasians. However, insulin secretory defect rather than insulin resistance has been speculated and demonstrated to be a more important factor in the development of type 2 diabetes in other ethnic groups. Thus, we undertook this study to investigate the initial abnormality of glucose intolerance in Korean subjects. METHODS: 374 Korean subjects were stratified according to the World Health Organization criteria (normal glucose tolerance [NGT], n = 128; impaired glucose tolerance [IGT], n=128; diabetes, n=118) and subdivided further into the two groups; non-obese (BMI < 25 kg/m2) and obese group (BMI 25 kg/m2). Insulinogenic index (the ratio of the increment of insulin to that of plasma glucose 30 min after glucose load) was used as an index of early-phase insulin secretion. AUC insulin (area under the insulin curve during OGTT) was used as an index of total insulin secretion. Insulin resistance was assessed by HOMA (R), the R value of the Homeostasis model. RESULTS: Insulinogenic index decreased significantly in IGT compared with that in NGT in both non-obese and obese groups, respectively. There was no significant difference in AUC insulin and HOMA (R) between NGT and IGT group. WhereasAUC insulin showed its peak level in the range of IGT (7.7~9.9 mmol/L), insulinogenic index showed the peak level in the range of NGT (5.6~7.7 mmol/lL and decreased progressively with increase of plasma glucose 120 min value. CONCLUSION: Early-phase insulin secretory defect might be the initial abnormality in the development of IGT from NGT in both non-obese and obese Korean subjects.
The Clinical Significance of Anthropometric Measurements of Obesity in Type 2 Diabetics.
Young Sun Choi, Si Hyung Park, Bo Wan Kim
Korean Diabetes J. 2000;24(3):365-374.   Published online January 1, 2001
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AbstractAbstract PDF
BACKGROUND
It is well demonstrated that obesity is associated with many chronic disorders like type 2 diabetes, dyslipidemia, and hypertension. In Korea, there are numerous reports revealed the possible relationship between anthropometric indices of obesity and the aformentioned disorders in nondiabetic healthy population, but the reports in type 2 diabetics are limited. Therefore, in this study we evaluated the relationship among the anthropometric indices and fasting insulin, C-peptide, and serum lipid levels in recently diagnosed type 2 diabetics. METHOD: A total of 160 type 2 diabetics were recruited from the out-patient department of endocrinoloy of Kyungpook National University Hospital from March to June 1999. The following subjects were excluded from the study: duration of type 2 diabetes > 5year, receiving of insulin or lipid lowering agents. For all participants, body mass index, percent of body fat, waist and hip circumference were measured. Fasting plasma insulin, C-peptide, blood glucose, HbA1c, total cholesterol, triglyceride and high-density lipoprotein cholesterol were also assayed. Partial correlation analysis was used to test the relationships between anthropometric indices and laboratory data. RESULTS: 1) In male subjects, fasting plasma insulin level was not significantly correlated with any of the anthropometric variables. However, in female subjects, fasting plasma insulin level was significantly correlated with body mass index, percent of body fat, and waist circumference. 2) Fasting plasma C-peptide level significantly correlated to all of the anthropometric variables in female patients, but in males only significant association was seen between fasting plasma C-peptide level and waist circumference. 3) Total cholesterol level was significantly correlated with percent of body fat and waist/hip ratio in male andfemale subjects respectively. CONCLUSION: Anthropometric indices of abdominal obesity appear to be correlated with insulin production and lipid changes in recently diagnosed type 2 diabetics. Waist circumference is a more useful predictor of hyperinsulinemia than waist/hip ratio in both sexes.
The Effect of Ginkgo Biloba Extract on Diabetic Peripheral Neuropathy - A 12 week, randomized, placebo-controlled, double-blind trial -.
Kyung Mook Choi, Dong Rim Kim, Nan Hee Kim, Sei Hyun Baik, Dong Seop Choi
Korean Diabetes J. 2000;24(3):375-384.   Published online January 1, 2001
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AbstractAbstract PDF
BACKGROUND
In the pathogenesis of diabetic neuropathy, metabolic derangement and ischemic damage have been considered as the major possible mechanisms. Ginkgo biloba extract was known to improve microcirculation by its vasodilator and antiplatelet effects, and used for peripheral and cerebral circulatory disorder. It also acts as free radical scavenger and inhibits oxidative damage. Thus, in this study we evaluate the effects of Ginkgo biloba extract on symptoms and nerve conduction study in patients with diabetic peripheral neuropathy. METHODS: In this study, over 3 months period, we recruited a total of 33 type 2 diabetic patients with peripheral neuropathy. Nineteen patients were randomly assigned to receive placebo, and fourteen patients to receive Ginkgo biloba extract (40 mg tid) for a duration of 12 weeks. We measured fasting blood glucose, postprandial 2 hour blood glucose levels, glycosylated hemoglobin and the lipid profiles. Clinical evaluation included neuropathy symptom score and nerve conduction study, and it was performed before and after the treatment. RESULTS: During the treatment, fasting blood glucose, postprandial 2 hour blood glucose, glycosylated hemoglobin and the lipid profiles were not significantly changed. Furthermore, no significant changes of neuropathy symptom score were observed during the treatment period. However, in Ginkgo biloba extract treatment group, motor nerve conduction velocities of median and ulnar nerve were improved significantly when compared to the placebo group. CONCLUSION: With the 12 weeks Ginkgo biloba extract treatment, we observed some improvement of nerve conduction velocity without any serious side effect.
Evaluation of 25% and 50% -75 gm Oral Glucose Tolerance Test - Animal and Clinical Pilot Study: Emphasis on Glucose Kinetics and Preference Evaluation.
Nam Han Cho, Eun Gyoung Hong
Korean Diabetes J. 2000;24(3):385-392.   Published online January 1, 2001
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AbstractAbstract PDF
BACKGROUND
Oral glucose tolerance test to diagnose diabetes was first introduced by Jarney and Isaacson in 1918. This method was advocated because of its diagnostic accuracy and also provides the two hours glucose changing pattern. Twenty-five percent (296 mL) glucose solution has been used as the standard diagnostic method in Korea. However, large volume of the solution frequently cause vomiting during the tests. Thus, 50% solution (150 mL) was recently introduced, but the potential difficulty of gastric emptying caused by its hyperosmolarity, and degree of diagnostic accuracy was questioned. Therefore, in this study, we evaluated two type of solutions by comparing the followings in both an animal and human models: (1) glucose changing pattern during the two hour oral glucose tolerance test, (2) Preference evaluation in human model. METHODS: Fifteen male Sprague-Dawley rats and 15 human subjects underwent 2 hours OGTT after 10~14 hours fasting. Two grams glucose per kg body weight was feed to the SD rat. In human, 75 gm glucose in 296 mL (25%) and 150 mL (50%) glucose solution was ingested at two different time, but testing was done within 24 hours a part. Five blood samples (fasting, 30, 60, 90, and 120 minutes) were collected and separated for serum. Glucose was assayed using YSI 2300-STAT (Yellow Springs Instrument Co., Ohio, USA) by glucose oxidase method. RESULTS: In animal study, despite the lower fasting glucose level, 30, 60 and 120 minutes glucose level was higher in 50% solution when compared to the 25% but the mean values were not statistically different. The glucose area under the curve (GAUC) in 50% was higher than 25% but not statistically different. The peak glucose level was observed at 60 minutes in both solutions. In human study, although mean values were not statistically different, all glucose values except 30 minutes were higher in 50% solution. Furthermore, GAUC was not statistically different between the two solutions. In preference test, the study subjects significantly (p<0.05) preferred the 50% solution as more favorable amount for the test. No differences in the tolerable level of sweetness, level of thirsty after ingestion, nausea, vomiting, head and stomachache was observed. CONCLUSION: In this study, we found that the gastro-intestinal glucose kinetics of the 25% and 50% glucose solution used during the OGTT was very similar in both an animal and human model. Furthermore, the preference evaluation showed favorable results in 50% solution. The use of 50% solution reconcile vomiting problem during the test but the same diagnostic accuracy was preserved. Therefore, 50% solution merits its scientific value as the diagnostic solution, and hope to contribute to favor the OGTT for diagnosis of diabetes mellitus in the future.

Diabetes Metab J : Diabetes & Metabolism Journal