Background: Acute gastroenterophathy usually caused by the Rota virus for children under 5 years old. Objectives: To present various types of data on epidemiology of ROTA virus diarrhea in Ho Chi Minh city from 12/2006-11/2007. Material and method: The data were collected from 500 stool specimens of diarrhea diagnosed chilren hosptalised at Thuy Dien Pediatric hospital 1, Ho Chi Minh city from December/2006 to November /2007. Results:There were 322 rotavirus-positive specimens, representing 64.4%. The proportions of monthly distribution of cases with diarrhea due to rotavirus were 90.1%, 54.39%, 85.37%, 74.51%, 72.92%, 41.67%, 26.67%, 58.33%, 79.31%, 52.63%, 69.05% and 57.78%, respectively. The numbers of rotavirus-positive cases in male and female were 216 (65.26%) and 106 (62.72%), respectively. The proportions of Rota virus positive children compared to total number of diarrheal cases with age 0-3, 3-6, 6-12, 12-24, 24-36 and over 36 months were 2.80%, 7.76%, 40.06%, 40.68%, 5.28% and 3.42%, respectively.\r\n', u'The results of typing identification indicated that the phenotypes of 98 among 100 specimens were identified (98%) in which there were sixty-one specimens of G1P8 (61%), one specimen of G2P8 (1%), fourteen specimens of G3P8 (14%), four of specimens of G4P8 (4%), eighteen specimens of GmixedP8 (18%). There were only two specimens of GnontypeableP8 (2%). Conclusion: Further studies should be carried out to clear this issue.\r\n', u'
Rotavirus ; gel type.

BACKGROUND: Polymerase chain reacation (PCR)-based methods have been described for rapid detection and identification of Candida spp. Multiplex PCR assay was developed using internal transcribed spacers and topoisomerase II gene for the accurate identification of Candida species. METHODS: We designed Dual Specificity Oligo (DSO) primers for multiplex PCR. Multiplex PCR was followed by agarose gel electrophoresis to test 8 type strains (C. albicans, C. parapsilosis, C. glabrata, C. tropicalis, C. krusei, C. guilliermondii, C. lusitaniae, C. dubliniensis) and 96 clinical isolates (C. albicans 51 isolates, C. parapsilosis 10 isolates, C. glabrata 10 isolates, C. tropicalis 9 isolates, C. krusei 6 isolates, C. guilliermondii 5 isolates, C. lusitaniae 5 isolates) of Candida spp. RESULTS: With multiplex PCR using DSO primers, the eight Candida type strains each could be easily differentiated and all 96 clinical isolates were identified as the same species as were identified by the conventional method. CONCLUSION: Multiplex PCR followed by electrophoresis can be useful for the simple and rapid identification of Candida species in routine laboratories.
Candida* ; DNA Topoisomerases, Type II ; Electrophoresis ; Electrophoresis, Agar Gel ; Multiplex Polymerase Chain Reaction* ; Sensitivity and Specificity

We report a case of type III hyperlipoproteinemia which is called a broad-beta disease. A 53 year old female patient visited our clinic for the evaluation of multiple yellowish papules on extremities and eyelids. The patient showed various types of xanthoma includiiig eruptive, tuberous, tendinous xanthomas and xanthelasma palpebrarum, xanthoma striatum palmare. The blood chemistry revealed a marked elevstion of cholesterol and triglycerides and agarose gel electrophoresis showed a single peak at prebeta and beta portion without separation. On histopathologic studies, typical foam cells were showen.
Chemistry ; Cholesterol ; Electrophoresis, Agar Gel ; Extremities ; Eyelids ; Female ; Foam Cells ; Humans ; Hyperlipoproteinemia Type III* ; Hyperlipoproteinemias ; Middle Aged ; Triglycerides ; Xanthomatosis

Among the fifty-three clinical isolates of Haemophilus influenzae, nineteen isolates including eight isolates of each biotype I-VIII, six of serotype b (Hib) strains and five of nontypeable strains were characterized by SDS-PAGE about outer membrane protein (OMP), restriction enzyme analysis (REA) and rRNA gene restriction pattems. OMP patterns showed to common band patterns in each H. influenzae isolate. Based on the two major proteins, 31KDa-38KDa, isolated strains were classified into 7 subtypes. In the OMP patterns about biotype and serotype, the specific pattern of each biotype was not distinguishable, but all of the serotype b strains were shown identical unique pattern, therefore it made distinctive difference with nontypeable strains. The digested genomic DNAs with EcoRI were identical result with rRNA gene restriction. It was more subdivided into 10 ribotypes. The most common ribotype I and serotype 1 accounted for 6 strains (31.6%) and 7 strains (36.8%) of the 19 clinical isolates, respectively. Hib isolates that were both OMP subtype 1 and ribotype I accounted for 2 strains (10.5%). In the epidemiologically unrelated strains, the putative association between the subtypes could not be confirmed. According to these results, the three methods were discriminatory and appropriate techniques for epidemiological studies of H. influenzae.
DNA ; Electrophoresis, Polyacrylamide Gel* ; Genes, rRNA* ; Haemophilus influenzae type b ; Haemophilus influenzae* ; Haemophilus* ; Influenza, Human ; Membrane Proteins ; Restriction Mapping* ; Ribotyping

Cartilage is commonly used autogenous material for aesthetic and reconstructive surgery and major donor sites of cartilage are ear, nasal septum, and rib. As the cartilage correlates with ossification and can be used for joint reconstruction. Many growth factors influencing growth and differentiation of chondrocytes have been reported, and matrix composition produced by chondrocytes may vary in types and quantity according to culture duration. Initially the chondrocytes in culture aggregate, then secrete type I collagen. Type II collagen is produced during differentiation process, and synthesis of type X collagen is the last step. In this study, chondrocytes were isolated from ear cartilage of the New Zealand white rabbit weighing 400 gm. We performed high density culture using penicylinder and pellet method. The cells were polygonal in morphology and viable under the inverted microscope. This experiment was designed to evaluate the effect of IGF-I, TGF- p, and b- FGF on the synthesis of collagen in chondrocyte culture. Optimal concentration of growth factors was determined using H-thymidine incorporation into DNA. After the addition of optimal concentration of each growth factors in experimental groups, the uptake of H-proline was measured. Only IGF-I showed a statistically significant increase of collagen synthesis. We observed how subtypes of collagen were influenced by growth factors in two culture methods and by differing the addition timing of growth factors. SDS-PAGE was adopted for subtyping of collagen. All subtypes of collagen were found in both culture methods and all growth factors facilitated the production of type II and type X collagen and may be devoted to the differentiation of chondrocytes. Immunohistochemical staining for type I, and type II collagen was examined to confirm the above result. In pellet culture, type II collagen was stained densely in response to the addition of three kinds of growth factors. The results of penicylinder culture showed similar outcome to those from pellet cultured group. From the above results, we concluded as follows; First, IGF-I generally influence the synthesis of type I and II collagen. Second, TGF beta increased the synthesis of collagen. Third, b-FGF increased the synthesis of type II and type X collagen. We concluded that IFG-I is the only growth factor which is effective regardless of culture duration and method. TGF- beta and b-FGF, which are potent mitogen, facilitate the secretion of collagen.
Cartilage ; Chondrocytes* ; Collagen Type I ; Collagen Type II ; Collagen Type X ; Collagen* ; DNA ; Ear ; Ear Cartilage ; Electrophoresis, Polyacrylamide Gel ; Humans ; Insulin-Like Growth Factor I ; Intercellular Signaling Peptides and Proteins* ; Joints ; Nasal Septum ; New Zealand ; Ribs ; Tissue Donors

Cartilage is one of the most commonly manipulated tissue in esthetic and reconstructive surgery. Cartilage has an important role in longitudinal bone growth. Anabolic hormones and locally produced peptide growth factors are known to influence this process Matrix composition changes through proliferation, maturation, and differentiation of chondrocytes, and endochondral ossification thereafter. Defined cartilage matrix is synthesized during the maturation of chondrocytes where the major change is the increment of type II collagen. Variable sulfated mucololysaccharides and hyaluronic acid are also synthesized during this maturation. IGF-I(insulin like growth factor-I), so called somatomedin C, is a prominent growth factor in serum. IGF-I is known to be involved in long growth. IGF-I is affected by pituitary growth hormone. There are few studies done on IGF-I effect in cartilage matrix formation and possible changes of collagen subtypes. This experiment was designed to see the IGF-I effect on the colagen synthesis of cultured chondrocytes. Optimal concentration of IGF-I for the experiment was determined using H3-thymidine incorporation into DNA. The IGF-I effect on collagen synthesis was studied using H3-proline. The IGF-I effect on the synthesis of subtypes of collagen was studied using SDS-PAGE and immunocytochemical staining. Chondrocytes were isolated from the ears of New Zealand white rabbit and cultured in 2 X 10(5) cells/300 microgram density. IGF-I increased DNA synthesis, and optimal concentration of IGF-I was determined by dose-relationship curve as 10ng/ml. Collagen synthesis was increased by IGF-I. Type II collagen was increased on SDS-PAGE with IGF-I and this gel electrophoresis showed type X collagen, also. The increase in type II collagen was confirmed with immunocytochemical staining, the reaction becoming stronger with the addition of IGF-I. Type I collagen was not changed with IGF-I on immunocytochemistry. We conclude that IGE-I is an important modulator influencing not only proliferation and maturation but also terminal different-iation of chondrocytes.
Bone Development ; Cartilage ; Chondrocytes* ; Collagen Type I ; Collagen Type II ; Collagen Type X ; Collagen* ; DNA ; Ear ; Electrophoresis ; Electrophoresis, Polyacrylamide Gel ; Growth Hormone ; Hyaluronic Acid ; Immunohistochemistry ; Insulin-Like Growth Factor I* ; Intercellular Signaling Peptides and Proteins ; New Zealand

BACKGROUND: Trichophyton rubrum is the most common dermatophyte isolated from human and has ability to invade the tissues such as stratum comeum, nail and hair. The potential role of proteinases as virulence factors of F rMSrMm has been discussed at length. OBJECTIVE: As a first step towards assessing its virulence role, we report on the purification and characterization of proteinase from T. rubrum isolate culture filtrates. METHODS: An extracellular serine proteinase has been purified from culture filtrates of Trichophyton rubrum HP-9 by ultrafiltration, gel filtration chromatography, and affinity column chromatography. Azocoll and keratin azure were employed as the substrates of enzyme activities. Peak of proteolytic activity was analyzed by gelatin co-polymerized gel electrophoresis. RESULTS: The molecular weight of the purified enzyme was approximately exhibited to 14.0 kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The optimum pH and molality of 14.0 kDa proteinase activity was 6.0 and 100mM, respectively. The activity was inhibited by serine proteinase inhibitor, phenylmethylsulfonyl fluoride (PMSF). The proteinase degraded gelatin, collagen type VI, and keratin from human epidermis but not hemoglobin. CONCLUSION: The 14,000 Mr extracellular serine proteinase purified from T. rubrum NP-9 culture filtrates has neutral pH optimum 6.0 and activities against gelatin, collagen type VI, and keratin.
Arthrodermataceae ; Chromatography ; Chromatography, Gel ; Collagen Type VI ; Electrophoresis ; Electrophoresis, Polyacrylamide Gel ; Epidermis ; Gelatin ; Hair ; Humans ; Hydrogen-Ion Concentration ; Molecular Weight ; Peptide Hydrolases ; Phenylmethylsulfonyl Fluoride ; Serine Proteases ; Sodium Dodecyl Sulfate ; Trichophyton* ; Ultrafiltration ; Virulence ; Virulence Factors

BACKGROUND: Low-density lipoprotein (LDL) cholesterol has been shown to be a major risk factor for coronary artery disease (CAD) in animal studies, clinical trials, and observational epidemiologic studies. It has a hydrated density of 1.019 to 1.063 kg/L, a diameter of 20 to 30 nm, and displays beta-mobility on paper or agarose gel electrophoresis. With technique such as density gradient ultracentrifugation and gel electrophoresis, it is possible to separate lipoproteins accurately on the basis of their density, charge, and particle size. Further, it has been shown that a smaller LDL is associated with an increased risk of coronary artery disease, even when total cholesterol level is only slightly raised. The aim of this study was to analyze LDL particle size distribution in patients with angiographically confirmed coronary artery disease and in control subjects, using nondenaturating gradient polyacrylamide gel electrophoresis, and to investigate the relationship between LDL particle size and the other traditional coronary risk factors. METHODS: Baseline characteristics such as age, sex, body mass index, history of hypertension or NIDDM, smoking habits, and plasma lipoprotein profiles were obtained in 33 and 27 subjects with and without CAD angiographically confirmed, respectively. We determined LDL peak particle diameter (LDL-PPD) using nondenaturating gradient polyacrylmide gel electrophoresis in CAD and control group. 4% to 12% polyacrylamide gradient gels were used for this assay, and the diameters of LDL subclass peaks were calculated by comparison with a standard calibration curve. This procedure permits the assignment according to distribution of particle diameters as exhibiting pattern A, B, or INT. RESULTS: Traditional coronary risk factors (age, sex, body mass index, history of hypertension or NIDDM, smoking habits, plasma triglyceride, HDL-cholesterol, and total to HDL-cholesterol ratio) were found to be significantly different between two groups, except the plasma total cholesterol and LDL-cholesterol. The mean value of LDL-PPD in patients with CAD was significantly lower than that in control subjects (26.110.4 nm versus 27.011.9 nm, p=0.006). LDL-PPD showed relatively strong associations with plasma triglyceride (r= - 0.536, p<0.01), HDL-cholesterol (r=0.497, p<0.01), and total to HDL-cholesterol ratio (r= - 0.516, p<0.01), but showed no relation to total cholesterol (r= - 0.168) or LDL-cholesterol (r= - 0.028). CONCLUSION: These results suggest an association between small LDL and the presence of CAD and also suggest that LDL-PPD may be associated with the plasma lipid levels.
Animals ; Body Mass Index ; Calibration ; Cholesterol ; Coronary Artery Disease* ; Coronary Vessels* ; Diabetes Mellitus, Type 2 ; Electrophoresis ; Electrophoresis, Agar Gel ; Electrophoresis, Polyacrylamide Gel ; Epidemiologic Studies ; Gels ; Humans ; Hypertension ; Lipoproteins* ; Particle Size* ; Plasma ; Risk Factors ; Smoke ; Smoking ; Triglycerides ; Ultracentrifugation

The amino group PEGylation of rhIFNomega with monomethoxy polyethylene glycol succinimidyl succinate (mPEG-SS, 20 000) was investigated, and the modified mixture was separated and purified by ion exchange chromatography and gel filtration chromatography. Under the optimized purification conditions, the average content ofmono PEG-rhIFNomega in the collect liquid reached 182 microg x mL(-1). The average purified yield of mono PEG-rhIFNomega exceed to 22%, and the purity of mono PEG-rhIFNomega was greater than 98% by SDS-PAGE and RP-HPLC. Relative molecular mass of mono PEG-rhIFNomega was 43 790 detected by MALDI-TOF MS. The apparent molecular mass measured by SDS-PAGE was about 60 810. The purified PEG-rhIFNomega has the characteristics of typical PEGylated protein. Activity reservation rate of mono PEG-rhIFNomega was 15.0%, while the antigenicity decreased by at least 64 folds. In addition, the acid stability, thermal stability and stability in serum and trypsin solution of mono PEG-rhIFNomega were markedly better than those of the rhIFNomega. The pharmacological properties of mono PEG-rhIFNomega were significantly improved. The prepared PEG-rhIFNomega might be developed to a novel safe and long-acting interferon.
Animals ; Antigen-Antibody Reactions ; Chromatography, Gel ; Chromatography, High Pressure Liquid ; Chromatography, Ion Exchange ; Drug Stability ; Electrophoresis, Polyacrylamide Gel ; Interferon Type I ; chemistry ; immunology ; Molecular Weight ; Polyethylene Glycols ; chemistry ; Rabbits ; Recombinant Proteins ; chemistry ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Fibronectin is involved in various cellular activities such as adhesion, proliferation and migration as a substratum. Since the metabolic turnover of fibronectin is much slower than other cellular components, it may be affected by the oxygen free radicals produced in the aging process. Hydroxyl radicals are known to be generated during inflammation and aging process. The purpose of this study is to investigate the influence of fibronectin treated with hydroxyl radicals generated by the H2O2-Cu2+ systems on osteoblast. The bone nodule formation, alkaline phosphatase activity, cell proliferation study and secretion of type I collagen were compared among non-coated, fibronectin-coated and hydroxyl radical treated fibronectin-coated groups. SDS-PAGE and Western blot analysis showed that fibronectin was degradated with hydroxyl radical treatment. The bone nodule formation in the fibronectin-coated group was significantly greater than that in non-coated or hydroxyl radical treated fibronectin-coated group. Alkaline phosphatase activity and cell proliferation were reduced in hydrokyl radical treated fibronectin coated group. In hydrosyl radical treated fibronectin-coated group, mRNA level of type I collagen by RT-PCR was also decreased. This study suggested that fibronectin played important roles in osteoblast activity and fibronectin damaged by the hydrokyl radicals reduced the bone formation of osteoblast through the inhibition of cell proliferation, differentiation and calcification processed.
Aging ; Alkaline Phosphatase ; Animals ; Blotting, Western ; Cell Proliferation ; Collagen Type I ; Electrophoresis, Polyacrylamide Gel ; Fibronectins* ; Free Radicals ; Hydroxyl Radical ; Inflammation ; Osteoblasts* ; Osteogenesis ; Oxygen ; Rats* ; RNA, Messenger