Effect of Human Parathyroid Hormone on Hematopoietic Progenitor Cells in NOD/SCID Mice Co-Transplanted with Human Cord Blood Mononuclear Cells and Mesenchymal Stem Cells

PURPOSE: We evaluated the effect of human parathyroid hormone (hPTH) on the engraftment and/or in vivo expansion of hematopoietic stem cells in an umbilical cord blood (UCB)-xenotransplantation model. In addition, we assessed its effect on the expression of cell adhesion molecules. MATERIALS AND METHODS: Female NOD/SCID mice received sublethal total body irradiation with a single dose of 250 cGy. Eighteen to 24 hours after irradiation, 1x107 human UCB-derived mononuclear cells (MNCs) and 5x106 human UCB-derived mesenchymal stem cells (MSCs) were infused via the tail vein. Mice were randomly divided into three groups: Group 1 mice received MNCs only, Group 2 received MNCs only and were then treated with hPTH, Group 3 mice received MNCs and MSCs, and were treated with hPTH. RESULTS: Engraftment was achieved in all the mice. Bone marrow cellularity was approximately 20% in Group 1, but 70-80% in the hPTH treated groups. Transplantation of MNCs together with MSCs had no additional effect on bone marrow cellularity. However, the proportion of human CD13 and CD33 myeloid progenitor cells was higher in Group 3, while the proportion of human CD34 did not differ significantly between the three groups. The proportion of CXCR4 cells in Group 3 was larger than in Groups 1 and 2 but without statistical significance. CONCLUSION: We have demonstrated a positive effect of hPTH on stem cell proliferation and a possible synergistic effect of MSCs and hPTH on the proportion of human hematopoietic progenitor cells, in a xenotransplantation model. Clinical trials of the use of hPTH after stem cell transplantation should be considered.

Association study of LEP and LEPR gene polymorphisms and the development of type 2 diabetes mellitus in Korean women with a history of gestational diabetes / 대한산부인과학회지

OBJECTIVE: The history of gestational diabetes (GDM) is a high risk for the development of type 2 diabetes mellitus (T2DM). The purpose of this study is to investigate the genetic association of LEP and LEPR gene polymorphisms and the development of T2DM in Korean women of history of GDM. METHODS: Women diagnosed as GDM during pregnancy from January 1992 to December 2002 were recruited. Those women with a T2DM at the time of study were classified as T2DM positive group, and without T2DM, as T2DM negative group. 2 genes (LEP and LEPR genes) and 8 SNPs (LEP-632G>A, +4950G>A, +4998A>C, and LEPR-141013T>C, -186A>G, +5193G>A, +7187A>C, +27265A>G) were selected. The TaqMan assay for genotyping and the statistical analysis for phenotypic and genetic factors between 2 groups were analyzed. RESULTS: A total of 54 women, T2DM positive (n=20) and T2DM negative (n=34) were enrolled. At the time of diagnosis of GDM, HbA1c, 50 g and 100 g oral glucose tolerance test, and insulin level were significantly associated between T2DM positive and negative groups (P<.05). In analysis of genetic risk to T2DM, the significant association related with any SNPs was not shown between T2DM positive and negative groups. CONCLUSION: In Korean women having past history of GDM, there was no relationship between 2 genes and the development to T2DM. To clarify a effect of candidate genes related with development of T2DM, there will need more samples and genes.

A De Novo Centric Fission of Chromosome 11 in a Patient with Recurrent Miscarriages

We report on a de novo centric fission of chromosome 11 in a healthy female referred for chromosome analysis due to recurrent miscarriages. Both fission products were mitotically stable. This centric fission of chromosome 11 appears to have no clinical significance for this patient other than recurrent miscarriages.

Identification of marker chromosomes by reverse painting fluorescence in situ hybridization and comparative genomic hybridization / 대한산부인과학회지

OBJECTIVE: Although marker chromosome is defined as an abnormal chromosome in which no part can be identified, derivative chromosomes with structural abnormalities of unknown origin are also called as marker chromosomes conventionally. The clinical significance of a marker chromosome is determined according to the origin of marker chromosome. In this study reverse painting fluorescence in situ hybridization (FISH), and comparative genomic hybridization (CGH) methods were employed to elucidate the origin of marker chromosomes in 5 clinical cases. METHODS: Reverse painting probes were generated from five copies of each marker chromosomes microdissected with micromanipulator, amplified with DOP-PCR, and labeled with fluorochromes. The probes were hybridized to normal metaphases. For CGH, normal control and patients' DNA were directly labeled with spectrum-red-dUTP and spectrum-green-dUTP by CGH nick translation kit, and hybridized to normal reference metaphases. The CGH images were captured with a computer controlled fluorescence microscope equipped with a CCD camera and analyzed by Cytovision workstation. RESULTS: Five marker chromosomes were identified as follows (1) derivative chromosome 15 inducing partial trisomy of 15pter->q21, (2) isochromosome of 18p causing 18p tetrasomy, (3) short arm of chromosome 5 causing 5p trisomy (4) small accessory chromosome originated from centromeric region of chromosome Xq11->q12 (5) der(17) with inverted duplication of the short arm of chromosome 17. In all cases the origin of each marker chromosomes were identified successfully with reverse painting FISH, and these results were concordant with the CGH profiles. CONCLUSION: Our results indicate that combined reverse painting FISH and CGH is a rapid, convinient and powerful tool to identify the origin of marker chromosomes and derivative chromosomes caused by various chromosome abnormalities such as translocation, duplication, deletion.

Chromosomal Aberrations in Korean Hepatocellular Carcinomas / 한국간담췌외과학회지

BACKGROUND/AIMS: The pattern of chromosomal gains and losses in HCC with hepatitis B in Korean patients is very complex and involves virtually every site in the genome. This study was done to know the chromosomal aberrations in hepatocellular carcinoma with HBV and relationship between these lesions and previously known oncogenes and tumor suppressor genes. METHODS: DNA changes in 23 hepatocellular carcinomas (HCC) associated with hepatitis B virus (HBV) were analyzed by CGH technique. RESULTS: Eighteen of the 23 cases showed genetic alterations. The remaining 5 cases showed no detectable aberrations. The losses of chromosome regions 17p (74%), 4q (57%), 16p (52%), 16q (48%), 8p (43%) and 13q (43%) were detected in the order of decreasing frequency. In cases of multiple losses of chromosomes, a combination of 17p, 16p, 16q, 4q, and 8p losses was found in 5 cases (30%). On the other hand, chromosomal gains occurred on 1q (65%), 8q (52%), 20p (48%) and 20q (43%) in the order of decreasing frequency. And the simultaneous genomic gains of these 4 chromosomes were found in 9 cases (40%). Moreover, the combination of 5 genomic losses (17p, 16p, 16q, 4q, & 8p) and 4 genomic gains (1q, 8q, 20p, & 20q) was observed in 4 cases (23%). CONCLUSIONS: The pattern of chromosomal gains and losses in HCC with hepatitis B in Korean patients is very complex and involves virtually every site in the genome. This indicates a high genomic instability. This could possibly be explained either as the result of random chromosomal changes during early development of tumor, or as the highly variable and random pattern of integration of HBV in the HCC. The hepatocarcinogenesis may be the result of cumulative effects rather than those orders of occurrence of those genomic changes. The sites of cellular DNA at which HBV integrates frequently undergo rearrangements, resulting in translocations, inverted duplications, deletions, and possibly recombinational events. But, CGH only detects changes of chromosomal copy number but could not identify translocations, inversions, and other aberrations of chromosome. The chromosomal analysis of HCC with HBV in Korean patients by CGH technique confirms the presence of complex and sporadic, but nonrandom genetic changes in the chromosome. In the future, more detailed oncogenic study could be carried out on the chromosomes which showed abnormal aberrations through this study.

CYP2D6 and NAT2 Polymorphism in HBV Associated in Hepatocellular Carcinoma Patients in Korea

PURPOSE: CYP2D6 and N-acetyltransferase (NAT2) are polymorphic enzymes which are expressed in the hepatocyte in a genotype-determined manner. They are known to be involved in the inactivation and activation of various mutagens and carcinogens, respectively. The activities of the two enzyme systems are associated with the genetic susceptibility of many human cancers. METHODS: This study was performed to determine the genotype frequencies of the two enzyme systems in primary hepatocellular carcinoma patients and healthy controls. One hundred healthy controls and 55 liver cancer patients were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: In the healthy controls, the CYP2D6 wild type allele frequency was 0.985 and the CYP2D6*4 frequency was 0.015. No CYP2D6 poor-metabolizer was detected. No significant differences were found in the hepatocellular carcinoma patients group. Among the controls, the frequencies of F, S1, S2 and S3 alleles of the NAT2 system were 0.725, 0.01, 0.14 and 0.125, respectively. The genotype frequencies were found to be 0.91 for the rapid acetylator and 0.09 for the slow acetylator. No significant differences were found in the hepatocellular carcinoma group. CONCLUSION: The distribution of CYP2D6 and NAT2 polymorphism is very unique in the Korean population, as characterized by the extremely low frequency of CYP2D6 poor-metabolizer and NAT2 slow acetylator. CYP2D6 and NAT2 polymorphisms did not seem to play an important role in the hepatic carcinogenesis in the Korean population.

A Case of Acute Promyelocytic Leukemia with 46, XX, del(5)(q23)/47, XX, del(5)(q23), +8 but Without (15;17) Translocation / 대한혈액학회지

Acute promyelocytic leukemia (APL/AML- M3) is a distinct subtype of acute myelogenous leukemia, which is characterized by unique morphologic, cytogenetic, molecular, and clinical features. In almost all APL patients, a characteristic t(15;17)(q22;q21) is found, resulting from the fusion of the PML gene and retinoic acid receptor alpha (RAR ) gene. This chromosomal translocation in APL may present variant translocations, and may be associated with secondary chromosomal abnormalities. The most frequent accompanying karyotypic aberration is trisomy 8 in APL. We are reporting a case of a 17-year-old woman who was diagnosed with APL. Cytogenetic study revealed that 46, XX, del(5)(q23)/47, XX, del(5)(q23), +8 chromosomal abnormality but without t(15;17). However, the presence of PML/RAR chimera was found with reverse transcriptase PCR. It is well known that the association of trisomy 8 on top of t(15;17) in APL cases. However, in our review, the mosaicism of del(5)(q23) with trisomy 8 in APL might be the first case. Whether this patient will behave the typical APL cases having good prognosis or not will be interesting to see.

Diagnosis of trichomoniasis by polymerase chain reaction

The clinical usefulness of polymerase chain reaction (PCR) for the diagnosis of trichomoniasis was evaluated in comparison with other conventional tests. PCR was used for specific detection of Trichomonas vaginalis by primers based on the repetitive sequence cloned from T. vaginalis (TV-E650). Between June 1996 and August 1997, 426 patients visited the department of obstetrics and gynecology, Hanyang University Kuri Hospital and were examined for trichomoniasis using wet mount examination, Papanicolaou (Pap) smear, culture and PCR. One hundred and seventy-seven patients (group A) visited with the symptoms of vaginal discharge and 249 patients (group B) visited for regular cervical Pap smear with no vaginal symptoms. From group A (n = 177), 3 infections (2.0%) were detected by wet mount, 6 infections (3.3%) by Pap smear and culture, and 17 infections (10.4%) by PCR. From group B (n = 249), 4 patients (1.6%) were found to have T. vaginalis by culture and 6 infections (2.4%) were detected by PCR. Therefore, in both groups, PCR for T. vaginalis showed a higher detection rate compared with conventional wet mount, Pap smear or culture. The detection by PCR was specific for T. vaginalis since no amplification was detected with DNAs from other protozoa and Candida albicans. The sensitivity and specificity of PCR were 100%. This method could detect T. vaginalis in vaginal discharge at a concentration as low as 1 cell per PCR mixture. These results indicate that PCR could be used as a specific and sensitive diagnostic tool for human trichomoniasis.

Microsatellite Instability and Its Clinicopathologic Significance in Korean Gastric Cancer Patients

BACKGROUND: Genomic instability has been reported as a novel mechanism of tumorigenesis. Microsatellites are short tandemly repeated nucleotide sequences throughout the human genome. Widespread somatic mutations in these sequences due to the loss or gain of one or more repeated units are termed as microsatellite instabilites (MI). MI have been found to be the result of numerous replication errors due to mutations in mismatched repair genes. Recently, MI have been recognized as the causes of the increased mutation rates of cancer cells. An elevated mutation rate manifested by MI may also affect various genes that are essential for normal cell function and growth, thus contributing to tumor initiation, promotion, and progression. We investigated the frequency of MI in a series of 44 gastric carcinomas in an attempt to clarify the role of these genetic alterations in gastric carcinogenesis and to see whether the cases with MI displayed any clinical significance and morphologic features and/or whether they showed distinctive relations with any clinicopathologic characteristics. METHODS: We analyzed 44 gastric carcinomas and paired samples from non-neoplastic mucosa of Korean patients who had undergone a gastrectomy. The samples were immediately frozen in liquid nitrogen and stored at 70oC until use. High molecular weight DNAs were isolated by standard methods only from cases whose were composed of more than 50% tumor cells. Ten loci of microsatellites were used in this study: D3S1766, D3S1339, D3S1029, D9S162, D9S171, INF-a, D11S925, D11S1818, D11S35, and D11S1284. The MI analysis was performed by Polymerace Chein Reaction (PCR) with 33P-labelled primers. PCR products were separated on 6% polyacrylamide gel containing 7M urea, and autoradiographed. H-E stained sections were used to review the pathologic features. The relationships between MI incidence and clinicopathologic findings were statistically tested. RESULT: Analysis of the 44 cases at the 10 loci of microsatellites allowed us to identify 19 instabilities (43%): 15 cases at 1 locus, 2 cases at 2 loci, and 2 cases at multiple loci. The incidence of MI was remarkably increased in cases with poor differentiation and node metastasis (p=0.067 and p=0.007, respectively). Comparing the histologic type (diffuse vs. intestinal type), the diffuse type of carcinoma had a higher incidence; however, there was no statistical significance (55% vs. 33%, p=0.108). There was no significant correlation between MI and other prognostic variables including location, tumor size, and distant metastasis. CONCLUSIONS: These results confirm that the mutator phenotype plays in a certain role gastric carcinogenesis. The gastric cancers with MI were correlated with the worse prognostic variables of poor differentiation and node metastasis, suggesting that MI appear to be useful as possible indicators of the worst prognosis and represent important genetic alterations associated with tumor progression in gastric carcinogenesis.

A study on the chromosomal aberrations in Korean intrahepatic cholangiocarcinomas with comparative genomic hybridization / 한국간담췌외과학회지

AIMS AND METHOD: Comparative genomic hybridization serves as a screening test for regions of copy number changes in tumor genomes. I have applied the technique to map DNA losses and gains in 13 cases of frozen cholangiocarcinomas. RESULTS: All of the 13 cases showed genetic alterations. Loss of short arm of chromosome 19 (92%) was the most common changes observed. 22q(62%), 1p(54%), 17p(54%) and 19q(54%) also showed nonrandom patterns of genomic losses with high frequencies. Among the genomic gains, 13q was revealed as the most common site (69%), and 8q (46%) and 12q (46%) also showed relatively high frequencies of genomic gains. Genomic amplifications were detected on 5p13, 10q21.1 and 18q11.3 in 3 different cases, respectively. CONCLUSION: This study represents the first analysis of intrahepatic cholangiocarcinomas by CGH, and it confirms the presence of nonrandom genetic changes occur in the pathogenesis of cholangiocarcinomas. These findings should lead to the characterization of new loci involved in cholangiocarcinoma pathogenesis.

Sister-Chromatid Exchanges in Lymphocytes of Medical Students Exposed to Formaldehyde / 대한산업의학회지

Sister-chromatic exchanges measured in the peripheral lymphocytes of 15 non-smoking medical students after exposure to formaldehyde during a 24-week anatomy class showed a small but significant (p=0.0468) increase when compared with samples obtained from the same individuals immediately before exposure. Mean frequencies of sister-chromatic exchange of cultured peripheral lymphocytes were 5.40+/-0.24 from the samples before exposure and 5.87+/-0.22 from the same samples after exposure. Breathing-zone air samples collected by formaldehyde monitoring kit with digital colorimeter (SKC) showed a mean concentration of 0.72+/-0.02 ppm formaldehyde.

Distinct Mutation of p53 Gene in Bladder Cancer of Smoking and Nonsmoking Patients / 대한비뇨기과학회지

PURPOSE: It is well known that smoking is one of the most important etiologic factor in bladder cancer and mutations of p53 tumor suppressor gene are the important step in carcinogenesis of urinary bladder. In this study, we investigated the difference in pattern and rate of p53 gene mutation between smoker and non-smoker MATERIALS AND METHODS: Of 26 bladder transitional cell carcinoma, 16 cases were smoker and 10 cases were non-smoker. We evaluated mutation of the p53 gene concentrated on axon 5 through 8, using polymerase chain reaction- single strand conformation polymorphism(PCR-SSCP) with radioisotope. RESULTS: 3 cases(18.7%) of 16 smoker were found to have p53 gene mutation, but none of 10 non-smoker was found. 2 of 3 cases of p53 gene mutation were found in exon 5 and 1 in exon 7. The pattern of p53 gene mutation was different in 3 cases. CONCLUSIONS: Although the more cases will be needed in this study, we think that a mutation of p53 in bladder cancer may be associated with cigarette smoking.

A Study on the p53 Mutations in Korean Breast Cancer Tissues / 대한암학회지

PURPOSE: The role of mutation of p53 gene on the carcinigenesis was studied since 1991. There were some relationships of p53 mutation and clinicopathologic factors. This sutudy was designed for the clinicopathologic and genetic factor relation in Korean breast cancer. MATERIALS AND METHOD: A retrospective study on the clinicopathologic findings such as age, menopausal status, TNM stage, histologic grade, estrogen receptor, DNA ploidy and S-phase fraction was camed out on 47 breast cancer tissues which had been resected at the Department of Surgery, Hanyang University Hospital. Forty-seven tissues were grouped into 3 groups. Group 1 was ductal carcinoma in situ, Group 2 was invasive ductal carcinoma without axillary lymph node metastasis and Group 3 was invasive ductal carcinoma with axillary lymph node metastasis. The numbers of tissues in each groups were 14, 15 and 18, respectively. Mutation screening on the p53 tumor suppressor gene was also performed with PCR-SSCP-direct sequencing method from the genomic DNA extracted from formalin fixed and paraffin-embedded pathologic tissue blocks. The results were as followings; RESULT: p53 mutations were detected in 12 cases(25.5%) of 47. In Group 1, 4 cases(28.6%) had mutations, and in Group 2, 5 cases(33.3%), and in Group3, 3 cases(16.7%). There was no significant differences in mutation rate between three groups. In 12 mutations detected, 6 cases were transition, 5 of which were missense mutation in coding sequences, and one of which was splicing mutation at acceptor site. One case was transversion and five cases were deletions or insertions of various lengths resulting in frameshift mutations. There was no statistically significant difference between groups and clinicopathologic factors except the strong relationship between the negative estrogen receptor and p53 mutation(p< 0.001). CONCLUSIONS: From the above findings, p53 gene could be considered to be inactivated at the all stage of multistep carcinogenesis processes. The nature of mutations and genetic background of Korean breast cancers may be somewhat different from those of Caucasians. And the p53 mutation status may be used as one of the useful prognostic factors in addition to the estrogen receptor status.

Microsatellite Polymorphisms in Intron 13 and Intron 22 of Factor VIII Gene / 대한산부인과학회잡지

The factor VIII gene comprises 26 exons spanning 185kb of DNA located at the distal end of the long arm of the X-chromosome, Defects in this gene cause hemophilia A, a bleeding disorder affecting 1/10,000 males. Linkage analysis is known as an efective method for the prenatal diagnosis and for the identification of carrier status. Several polymorphic markers had been studied to establish the diagnostic procedure for hemophilia A in Korea, and heterozygosity of 96% could be expected with 4 markers such as St14.1/Taq I, intron 18/Bcl I, intron 22/Xba I and DX13/Bal II. But in some families, above markers were not informative, and it was required another polymorphic markers should be applied for the diagnosis. Two recently identified microsatellite polymorphisms in intron 13 and intron 22 of FVIII gene were investigated to increase the heterozygosity and to diagnose previously uninformative families. Intron 13(CA)n repeats polymorphism showed 7 alleles with expected heterozygosity of 0.5336. Intron 22(CA)n(TC)n repeats polymorphism showed 4 alleles with expected heterozygosity of 0.5146. With the two microsatellite polymorphisms we could expect the heterozygosity of 0.6756. And we could successfully perform prenatal diagnosis previously uninformative family with intron 13 microsatellite polymorphism. With 4 polymorphisms detected by polymerase chain rection(intron 13 and intron 22 microsatellite polymorphisms, intron 18/Bcl I and St14.1 VNTR/Taq I), about 97% of hemophilia A family in Korea would be diagnosed by linkage analysis.

Analysis of p53 and Retinoblasoma(Rb) Gene Polymorphisms in Relation to Lung Cancer in Koreans / 결핵

BACKGROUND: The p53 and retinoblastoma(Rb) tumor suppressor genes are associated with the pathogenesis of several types of human cancer. Substantial. proportion of the primary lung cancers or cell lines have been reported to have the p53 and/or the Rb gene mutations. But so far there is no report on the analysis of the Rb gene polymorphism as one of the genetic susceptibility marker. This study was undertaken to establish the gene frequencies of the polymorphic genotypes of the p53 and Rb genes in Koreans to evaluate the possible involvement of these genotypes as a risk factor of lung cancer. METHODS: In this study 145 controls without previous and present tumor history and 128 lung cancer patients were subjected to analysis The two intragenic polymorphisms of the p53 gene(exon 4/AccII, intron 6/MspI) and one intron 17/XbaI polymorphism of the Rb gene were analysed by the method of polymersae chain react lion-restriction fragment length polymorphisms(PCR-RFLPS). The genotype of the intron 3/16 bp repeat polymorphism of p53 was determined by PCR and direct gel electrophoresis. RESULTS: There were no significant differences in the genotype distributions of the p53 gene between lung cart cert patients and controls. But heterozygotes(Arg/Pro) of the exon 4/AcclII polymorphisms were slightly over-represented than controls, especially in the Kreyberg type I cancer, which was known 13 be associated with smoking. The intron 3/16 bp duplication and the intron 6/MspI polymorphisms were in complete linkage disequilibrium. About 95% of the individuals were homozygotes of the common alleles both in the 16 duplication and MspI polymorphisms, and no differences were deteced in the genotype distributions between lung cancer patients and controls. Overall genotype distributions of the Rb gene polymorphisms between lung cancer patients and controls were not significantly different However, the genotype distributions in the Kreyberg type I cancer were significantly different from those of controls(p=0.0297) or adenocarcinomas(p=0.0008). It was noticeable that 73.4% of the patients with adenocarcinomas were heterozygotes(r1/r2) whereas 39.2% of the Kreyberg type I cancer were heterozygous at this polymorphisms. In the lung cancer patients, significant differences were a]so noted between the high dose smokers and low dose smokers including non-smokers(p=0.0258). The relative risk to Kreyberg type I cancer was significantly reduced in the individuals with the genotype of r1/r2(odds ratio=0.46, 95%C.I.=0.25-0.86, p=0.0124). The combined genotype distribution of the exon 4/AccII of the p53 and the intron 17 Rb gene polymorphisms in Kreyberg type I cancers were significantly different from dose of controls or adenocarcinomas. The highest odds ratio were observed in the individuals with the genotypes of Arg/pro and r2/r2(odds ratio=1.97, 95%C.I.=0.84-4.59) and lowest one was in the patients with Arg/Arg, r1/r2 genotype(odds ratio=0.54, 95%C.I.=0.25-1.14). CONCLUSION: The p53 and the Rb gene polymorphisms modulate the risk of smoking induced lung cancer development in Koreans. However, the exact mechanism of risk modulation by these polymorphism remains to be determined. For more discrete clarification of associations between specific genotypes and lung cancer risk, the evaluations of these polymorphisms in other ethnics and more number of patients will be needed.

Amplification of Cyclin D1 in Head and Neck Squamous Cell Carcinomas / 대한이비인후과학회지

BACKGROUND: Amplification of the chromosome 11q13 region has been observed in a variety of human cancers, including head and neck squamous cell carcinoma and carcinomas of breast, esophagus, lung, bladder, and liver. The chromosome 11q13 region has various putative oncogenes, of which cyclin D1 is most consistently amplfied and overexpressed. OBJECTIVES: To estabilsh the frequency and clinicopathologic correlations of cyclin D1 amplification in head and neck squamous cell carcinomas. MATERIALS AND METHODS: Fresh tissue samples were obtained from 26 patients with head and neck cancers undergoing surgery or biopsy. Amplification of cyclin D1 was evaluated in 26 head and neck squamous cell carcinomas by Southern blotting. The presence or absence of amplification was correlated with anatomic site, tumor stage, and differentiation pattern. RESULTS: Five tumors of 26(19%) showed a twofold to 4-fold amplification of cyclin D1 compared with beta-actin control prove. Amplified and nonamplified groups revealed no differences in anatomic primary site, stage, N stage, and pathologic differentiation. CONCLUSION: We showed a significant incidence of cyclin D1 amplification in head and neck squamous cell carcinomas, but cannot demonstrate an association with clinical presentation and pathologic findings.

Detection of the c-m c Oncogene Amplification in Ovarian Carcinomas by Differential Polymerase Chain Reaction

The amplification of c-myc oncogene was evaluated in 42 cases of ovarian carcinomas to correlate with clinical parameters. Using oligonucleotide primers, sequences from the c-myc exon-3 gene and from a control gene, tissue plasminogen activator (tPA), were amplified simultaneously by polymerase chain reaction (PCR). After the products of differential PCR (d-PCR) were electrophoresed, slot blot hybridization was performed, and hybridized with P32 dATP-labeled myc and tPA oligonucleotide probes and then autoradiographed. The signal intensities of the two products were quantitated by densitometry and the ratios of two products (c-myc/tPA) were measured. The ovarian carcinomas showed significantly increased amplification of c-myc oncogene Oligonucleoti compared to normal control group (p<0.05). 15 of 42 cases (35.7%) showed various degrees of the MYC gene amplification up to 27 folds in various histologic types of ovarian carcinomas. No significant differences of the MYC gene amplification according to histologic subtypes, tumor action) grades and clinical stages of ovarian carcinomas were present.

The CTG Repeat Polymorphisms of Myotonic Dystrophy (DM) Gene in Korean Population

Myotonic dystrophy(DM) is caused by the expansion of CTG trinucleotide repeat near the 3' end of the gene encoding for a member of protein kinase gene family (DMPK). The normal range of the CTG repeat was determined in 178 nomal individuals (141 unrelated individuals and 37 of 9 families) by polymerase chain reaction (PCR), polyacrylamide gel electrophoresis and silver staining method. And the expansion of the CTG repeats in a DM family was analyzed with Southern analysis. In normal population, the range of CTG repeat is between 5 and 34 and 19 different alleles were obserbed in that range, and (CTG)11-14 alleles were predominant. 4 members of an affected family showed the 0.5 - 2.0 kb size expansion of CTG repeats. In this study we could predict the incidence of DM in Korea as 1 in 20,000 and we could establish the diagnostic procedure for myotonic dystrophy.

A study on the patterns of expression of the DAZ and HSP genes in the testicular tissue of men with azospermia

In order to examine whether microdeletions on the Y chromosome exist or not and observe the aspects of expression of DAZ which is suggested to be essential in spermatogenesis in testicular tissue, tissues of 21 patients with azospermia were analyzed by using PCR methods and reverse transcription-PCR. The primers used for the analysis of the microdeletions on the Y chromosome were gene-specific. According to the results of the PCR with genomic DNA of the peripheral blood extracted from each patient, of the 21 men with azospermia 2 displayed microdeletions of the DAZ gene in the Y chromosome but none of HSP70A and HSP70B. And the reverse transcription-PCR of the RNA extracted from the testicular tissue of the patients gave results which found no amplified products of the mRNA of DAZ in the patients with microdeletions of that gene as expected, and confirmed patterns of expression of the mRNA of HSP70B in 5 whereas none could be seen with regard to HSP70A. Furthermore, the sperm of these 5 men were discovered t In accordance with the results previously described, one can see that the microdeletions of DAZ are associated with spermatogenesis and contemplate that HSP70B plays an important part in the maturation process of sperm. But it is considered that there is no correlation between the genes DAZ and HSP and since factors associated with the process of spermatogenesis are being continously discovered more studies on this should be advanced.

Molecular genetic study of St14.1(DXS52) TaqI RFLPs in Koreans for the diagnosis of hemophilia A / 대한산부인과학회잡지

No abstract available.