OBJECTIVEDespite the causes for melanin increase, the increased gene expression of TYR is a common pathological process. Based on this viewpoint, antisense-S-Oligo of TYR was designed and synthesized to regulate synthesis of melanin in order to explore the treatment for skin pigmentation.

METHODSThe cultured melanocytes were divided into 3 groups. The group 1 was treated with endothelin, group 2 treated with ultraviolet ray and group 3 was used as the control. In each group, the 5' antisense-S-Oligo, the 3' antisense-S-Oligo, the mixed antisense-S-Oligo of TYR or Dotap only was added. The melanin content and TYR gene expressions were examined.

RESULTSThe 5' antisense-S-Oligo, the 3' antisense-S-Oligo and the mixed antisense-S-Oligo significantly inhibited the increase of melanin content and TYR gene expression, which were caused by endothelin or ultraviolet ray treatment. Of the three treatments, the 3' antisense-S-Oligo showed the strongest effect.

CONCLUSIONAntisense-S-Oligo has significant regulating effects on TYR gene expression and melanin content. The 3' antisense-S-Oligo is more effective than the 5' antisense-S-Oligo.

3' Flanking Region ; genetics ; 5' Flanking Region ; genetics ; Endothelins ; pharmacology ; Gene Expression ; Melanins ; biosynthesis ; Melanocytes ; drug effects ; metabolism ; radiation effects ; Oligodeoxyribonucleotides, Antisense ; genetics ; pharmacology ; Tyrosine ; genetics ; metabolism ; Ultraviolet Rays

5' and 3' flanking region of ovine BLG were amplified from sheep genomic DNA according to the published whole sequence of ovine BLG and cloned to pGEM-T vector correspondently. By partially sequencing, the sequences of BLG 5' and 3' flanking were the same as that of publication completely. The recombinant structure used to direct exogenous gene especially to express in mammary gland was constructed by joining 4.2 kb 5' flanking with 2.1 kb 3' flanking. In order to assess the efficiency of BLG regulatory elements, green fluorescent protein (GFP) gene as a reporter was fused with BLG construct and transfected the mammary epithelial cells (TD47). Through observation under UV microscope and detection by fluorometer, it is demonstrated that the GFP has been successfully expressed in TD47 cell line. By virtue of direct observation and quantitative analysis, the BLG-GFP construct can be served as a model for the quick assessment of mammary gland expression construct.
3' Flanking Region ; genetics ; 5' Flanking Region ; genetics ; Animals ; Breast ; cytology ; Cell Line ; Cloning, Molecular ; Gene Expression Regulation ; Genes, Reporter ; Green Fluorescent Proteins ; Lactoglobulins ; biosynthesis ; genetics ; Luminescent Proteins ; biosynthesis ; genetics ; Sheep

Lipoprotein lipase (LPL) plays an essential role in the regulation of high-density lipoprotein cholesterol (HDLC) and triglyceride levels, which have been closely associated with cardiovascular diseases. Genetic studies in European have shown that LPL single-nucleotide polymorphisms (SNPs) are strongly associated with lipid levels. However, studies about the influence of interactions between LPL SNPs and lifestyle factors have not been sufficiently performed. Here, we examine if LPL polymorphisms, as well as their interaction with lifestyle factors, influence lipid concentrations in a Korean population. A two-stage association study was performed using genotype data for SNPs on the LPL gene, including the 3' flanking region from 7,536 (stage 1) and 3,703 (stage 2) individuals. The association study showed that 15 SNPs and 4 haplotypes were strongly associated with HDLC (lowest p = 2.86 x 10(-22)) and triglyceride levels (lowest p = 3.0 x 10(-15)). Interactions between LPL polymorphisms and lifestyle factors (lowest p = 9.6 x 10(-4)) were also observed on lipid concentrations. These findings suggest that there are interaction effects of LPL polymorphisms with lifestyle variables, including energy intake, fat intake, smoking, and alcohol consumption, as well as effects of LPL polymorphisms themselves, on lipid concentrations in a Korean population.
3' Flanking Region ; Alcohol Drinking ; Cardiovascular Diseases ; Cholesterol ; Cross-Sectional Studies ; Energy Intake ; Genotype ; Haplotypes ; Life Style ; Lipoprotein Lipase ; Lipoproteins ; Polymorphism, Single Nucleotide ; Smoke ; Smoking

Lipoprotein lipase (LPL) plays an essential role in the regulation of high-density lipoprotein cholesterol (HDLC) and triglyceride levels, which have been closely associated with cardiovascular diseases. Genetic studies in European have shown that LPL single-nucleotide polymorphisms (SNPs) are strongly associated with lipid levels. However, studies about the influence of interactions between LPL SNPs and lifestyle factors have not been sufficiently performed. Here, we examine if LPL polymorphisms, as well as their interaction with lifestyle factors, influence lipid concentrations in a Korean population. A two-stage association study was performed using genotype data for SNPs on the LPL gene, including the 3' flanking region from 7,536 (stage 1) and 3,703 (stage 2) individuals. The association study showed that 15 SNPs and 4 haplotypes were strongly associated with HDLC (lowest p = 2.86 x 10(-22)) and triglyceride levels (lowest p = 3.0 x 10(-15)). Interactions between LPL polymorphisms and lifestyle factors (lowest p = 9.6 x 10(-4)) were also observed on lipid concentrations. These findings suggest that there are interaction effects of LPL polymorphisms with lifestyle variables, including energy intake, fat intake, smoking, and alcohol consumption, as well as effects of LPL polymorphisms themselves, on lipid concentrations in a Korean population.
3' Flanking Region ; Alcohol Drinking ; Cardiovascular Diseases ; Cholesterol ; Cross-Sectional Studies ; Energy Intake ; Genotype ; Haplotypes ; Life Style ; Lipoprotein Lipase ; Lipoproteins ; Polymorphism, Single Nucleotide ; Smoke ; Smoking

BACKGROUND: Neurofibromatosis type 2 (NF2) is an autosomal dominant syndrome caused by the NF2 tumor suppressor gene. However, the NF2 mutation characteristics in Korean patients are not sufficiently understood. In this study, we conducted a comprehensive mutational analysis in 7 Korean NF2 patients by performing direct sequencing and gene-dosage assessment. METHODS: We analyzed all exons and flanking regions of NF2 by direct sequencing and screened the deletions or duplications involving NF2 by multiplex ligation-dependent probe amplification. RESULTS: Four novel NF2 mutations, including 2 splice-site mutations (c.364-1G>A and c.886-3C>G), 1 frameshift mutation (c.524delA), and 1 missense mutation (c.397T>C; p.Cys133Arg), were identified in our patients. No large deletion or duplication was identified in our series. Subsequently, we identified an abnormal splicing product by using reverse transcription-PCR and direct sequencing in 2 patients with a novel splice-site mutation. The missense mutation c.397T>C was predicted to have harmful effects on protein function. CONCLUSIONS: The detection rate of NF2 mutations in Korean patients (57%) is similar to those in other populations. Our results provided a greater insight into the mutational spectrum of the NF2 gene in Korean subjects.
3' Flanking Region/genetics ; 5' Flanking Region/genetics ; Adult ; Aged ; Amino Acid Sequence ; Asian Continental Ancestry Group/*genetics ; Child, Preschool ; Exons ; Female ; Frameshift Mutation ; *Genes, Neurofibromatosis 2 ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; *Mutation ; Mutation, Missense ; Neurofibromatosis 2/diagnosis/*genetics ; RNA Splice Sites ; Republic of Korea ; Sequence Analysis, DNA ; Young Adult

This study was conducted to assess the value of DNA diagnosis of gestational trophoblastic disease using polymerase chain reaction method. The 3 genetic loci including a variable number of tandem repeats regions were amplified by the PCR method on DNA of lymphocytes separated from the peripheral blood of the 53 uterine myoma patients. The distribution ranges of the APOB/VNTR, COL2A1/VNTR and MCT 118/VNTR were 691~850 bp, 651~720 bp and 501~720 bp respectively. The heterozygosity indices of the APOB/VNTR, COL2A1/VNTR and MCT 118/VNTR were 66.0%, 64.2% and 67.9% respectively. The author used the hypervariable 3' flanking region of the APOB/VNTR locus as target for DNA diagnosis of gestational trophoblastic disease. In 12 cases of hydatidiform mole, 1 case of invasive mole, and 1 case of choriocaricinoma, the target locus was amplified by the PCR method on DNA from lymphocytes of patients and their husbands, on DNA from the tisues. 10 cases of hydatidiform mole revealed DNA segments unique to the paternal APOB allele showing androgenesis. Two of theses androgenetic hydatidiform noles were heterozygous and the others were homozygous. A case of invasive mole showed normal genetic combination and a case of choriocarcinoma showed homozygosity. The heterozygous hydatidifomr moles as well as the homozygous ydatidiform moles were good in prognosis. The PCR method for targeting the APOB/VNTR appeared useful for the early diagnosis of hydatidiform mole.
3' Flanking Region ; Alleles ; Apolipoproteins B ; Choriocarcinoma ; Diagnosis ; DNA ; Early Diagnosis ; Female ; Genetic Loci ; Gestational Trophoblastic Disease* ; Humans ; Hydatidiform Mole ; Hydatidiform Mole, Invasive ; Leiomyoma ; Lymphocytes ; Minisatellite Repeats ; Polymerase Chain Reaction* ; Pregnancy ; Prognosis ; Spouses

OBJECTIVETo determine whether vitamin D receptor(VDR) gene start codon polymorphisms and 3'-end region polymorphisms exerted a combined influence on bone mineral density(BMD) in Han postmenopausal women in Beijing area.

METHODSThe VDR Fok I and 3'-end region genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism in 110 unrelated postmenopausal women. BMD was measured at the lumbar spine (L(2-4)), femoral neck(Neck), Ward's triangle(Ward's) and trochanter (Troch) using duel-energy X-ray absorptiometry.

RESULTSThe frequencies distribution of Fok I, Apa I, Bsm I and Taq I alleles in this cohort all followed the Hardy-Weinberg equilibrium. No significant association of Fok I, Apa I or Taq I genotype with BMD in postmenopausal women was found when these polymorphisms were considered independently, except for Bsm I genotype. When a combined analysis of VDR gene Fok I and 3'-end region polymorphisms was carried out, cross-genotyping Fok I and Apa I polymorphisms was significantly associated with BMD at the L(2-4) (P<0.001), and cross-genotype of Fok I and Taq I was also significantly associated with BMD at the Neck and Troch sites (P<0.05). However, cross-genotyping Fok I and Bsm I polymorphisms was not significantly associated with BMD. Cross-genotyping Apa I and Bsm I or Taq I polymorphisms was not associated with BMD in postmenopausal women, either.

CONCLUSIONAlthough Fok I polymorphisms of VDR gene were not significantly associated with BMD in postmenopausal women, VDR gene Fok I and 3'-region polymorphisms (Apa I and Taq I) had a combined effect on the BMD in postmenopausal women.

3' Flanking Region ; genetics ; Aged ; Analysis of Variance ; Bone Density ; physiology ; China ; Codon, Initiator ; genetics ; DNA ; genetics ; metabolism ; DNA Restriction Enzymes ; metabolism ; Female ; Gene Frequency ; Genotype ; Humans ; Middle Aged ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Postmenopause ; genetics ; physiology ; Receptors, Calcitriol ; genetics

OBJECTIVETo determine whether the muscle-specific glycogen-targeting regulatory subunit of the glucogen bound protein phosphatase 1 (PPP1R3) gene 5 bp deletion/insertion(D/I) within 3'-untranslated region ( 3'-UTR) polymorphism is associated with type 2 diabetes in Chinese Han population in Hefei region of Anhui province.

METHODSThe PPP1R3 gene 3'-UTR 5 bp D/I polymorphism was detected by polymerase chain reaction in 268 patients with type 2 diabetes and 106 normal controls.

RESULTS(1) The distributions of the frequency of three genotypes and two alleles of the PPP1R3 gene 5 bp D/I polymorphism showed no significant difference between the type 2 diabetic cases and the normal controls. (2) In both the cases and controls, there was no significant difference in age at onset, duration of disease, blood glucose, blood lipid profile, blood pressure, insulin sensitive index, body mass index, and waist hip ratio between the three genotypic groups(P 0.05). (3) The PPP1R3 gene 3'-UTR polymorphism in Chinese Han population in Hefei region of Anhui province was found to be similar to that in both Japanese population and Canadian population, and to be different from that in Piman Indians and the Caucasians in Sweden.

CONCLUSIONThe PPP1R3 gene 5 bp D/I within 3'-UTR polymorphism taking on genetic variation among the different races of mankind may not play a critical role in the development of type 2 diabetes mellitus in Chinese Hans of Hefei region in Anhui province.

3' Flanking Region ; genetics ; Aged ; Alleles ; Diabetes Mellitus, Type 2 ; enzymology ; genetics ; pathology ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Mutagenesis, Insertional ; Phosphoprotein Phosphatases ; genetics ; Polymorphism, Genetic ; Protein Phosphatase 1 ; Sequence Deletion

OBJECTIVETo study the polymorphism of c-Ha-ras 3' variable number of tandem repeat (VNTR) and its relationship with development of postmolar tumor.

METHODSThe cases of hydatidiform mole (HM) were retrospectively analyzed by amplified fragment length polymorphism-polyacrylamide gel electrophoresis. The DNA origin of HM was determined by comparison with the parents' DNA amplified results.

RESULTSAmong the samples from 15 cases, DNA from only paternal origin was found in 2 cases. DNA from both parents was in 13 cases, and of these 13 cases, 2 were found to be had balanced DNA origin, 11 had more DNA form paternal origin than DNA from maternal origin.

CONCLUSIONThe HM which has DNA from both parents origin and predominantly from paternal origin developed to postmolar tumor more frequently.

3' Flanking Region ; genetics ; DNA Mutational Analysis ; methods ; DNA, Neoplasm ; genetics ; Electrophoresis, Polyacrylamide Gel ; Female ; Genes, ras ; genetics ; Humans ; Hydatidiform Mole ; genetics ; Male ; Minisatellite Repeats ; genetics ; Nucleic Acid Amplification Techniques ; Polymorphism, Genetic ; Pregnancy ; Retrospective Studies ; Uterine Neoplasms ; genetics