Correlation Of Human Papillomavirus Infection and Postmenopausal Squamous Atypia in Cervical Cytology / 대한세포병리학회지

Postmenopausal squamous atypia (PSA) is a phenomenon characterized by cellular alterations mimicking condyloma in the uterine cervix of postmenopausal women. It is not associated with human papillomavirus (HPV) infection. The aim of this study is to correlate findings with HPV infection and the cytohistologic findings of PSA. Eighty-three smears from postmenopausal women, initially interpreted as ASCUS and low-grade squamous intraepithelial lesions(LSIL), were reviewed according to the criteria of PSA. Fifty-eight cases were subsequently reclassified as PSA. Forty cases categorized as PSA were available for HPV-DNA detection by a nested polymerase chain reaction. Eight of these 40 cases(20%) showed biopsy-proven LSIL lesions. The HPV-DNA was detected in 42.5%(17/40), compared to 25%(5/20) of control cases. The HPV-DNA detection rate of biopsy-proven LSIL was 62.5%(5/8). It has been concluded that cytologic differential diagnosis of PSA from LSIL is difficult due to because of poor histologic and viral correlation.

Association of Angiotensin-Converting Enzyme and Angiotensinogen Gene Polymorphisms with Preeclampsia

We tested the hypothesis that angiotensin-converting enzyme (ACE) and angiotensinogen gene polymorphism influence the incidence, development and outcome of preeclampsia. Subjects were recruited from 90 Korean patients with preeclampsia during pregnancy and 98 age-matched controls. After isolation of DNA, polymerase chain reactions (PCR) were carried out to detect polymorphism of the ACE and angiotensinogen. M235T and T174M genotypes of angiotensinogen were determined by digestion with restriction enzyme endonuclease Tth 111-I and NCo I, respectively. The frequency of DD genotype was significantly greater in preeclampsia (0.36) than in controls (0.14) (p<0.05). The frequency of D allele was 0.55 in preeclampsia and 0.40 in controls (p<0.05). There were no differences in the onset of preeclampsia and pregnancy outcomes according to the ACE genotypes. There was no difference in the frequency of a allele of angiotensinogen M235T between the groups (0.79:0.78 in preeclampsia : controls). The frequency of T allele of angiotensinogen T174M gene was slightly increased, but not significantly, in preeclampsia (0.11) than in controls (0.07). In a multivariate analysis, only ACE genotype was associated with the development of preeclampsia (beta=0.27, p=0.05). In conclusion, a molecular variant of ACE, but not angiotensinogen, gene is associated with preeclampsia in Korean women.

Identification of a mutation in the human raloxifene response element of the transforming growth factor-3 gene

The human transforming growth factor-3 (TGF-3) is an important cytokine to maintain bone mass by inhibiting osteoclast differentiation. Recently raloxifene response element (RRE), a new enhancer with a polypurine sequence for estrogen receptor (ER)-mediated gene activation, was identified on the TGF-3 gene. Functional analysis of the RRE-mediated pathway has shown that this would be an important pathway for bone preserving effect. We found a novel mutation in the RRE sequence by single-strand conformational polymorphism analysis in one of 200 Korean women. Cloning and sequencing revealed a heterozygote in which one allele had an insertion of 20 nucleotides (AGAGAGGGAGAGGGAGA GGG) between nucleotide +71 and +72 and a point mutation at nucleotide +75 (G-A transition), and the other allele had normal sequence. The insertion was a nearly perfect tandem duplication of the wild type DNA sequence. The bone mineral density of the affected woman was not much lower than that of age-matched controls. Transient transfection of the mutant allele showed no significantly different activity compared with that of the wild type allele. These observations suggest that the heterozygote variation of the RRE sequence seems not to be operative in determination of bone mass.

The Regulation of OPG/OCIF mRNA Epression by IL-1beta in Peripheral Blood Mononuclear Cells / 대한내분비학회지

BACKGROUND: Osteoprotegerin(OPG) is a soluble member of the tumor necrosis factor(TNF) receptor family and inhibits osteoclastogenesis by interrupting the cell-to-cell interaction between osteoblastic/stromal cells and osteoclast progenitors. OPG is expressed in many tissues including osteoblasts and may act on bone tissues in a paracrine and/or autocrine fashion. Futhermore, many cytokines and growth factors are known to influence the regulation of OPG expression in osteoblastic/stromal cells. The aims of the present study were to examine whether or not OPG was expressed in human peripheral blood mononuclear cells(PBMCs) and to investigate the effects of IL-1beta, which were known as potent osteotropic agents, on the regulation of OPG mRNA in PBMCs. METHODS: PBMCs were isolated by centrifugation over Ficoll-Hypaque density gradients from postmenopausal women and cultured in 6-well plates containing alpha-MEM supplemented with 5% FBS. The expression of OPG mRNA in PBMCs was observed by RT-PCR in adherent and nonadherent cells on culture plates. To observe the effect of OPG expression by IL-1beta, we measured the concentration of OPG mRNA by altering the concentration and incubation time of IL-1beta. The measurement of OPG mRNA was done by semi-quantitative PCR and indicated as OPG/GAPDH. RESULTS: OPG was expressed both in cells attached to the surface of culture plates and in non-adherent cells for the incubation of peripheral blood mononuclear cells. The effect of OPG mRNA by IL-1beta tend to increase in accordance with the length of incubation time and maximizes at 12 hours of incubation time and shows 1.2-3.5 times higher than the standard level at the concentration of 0.5ng/ml. However, the increased quantity in concentration varies according to individuals.] CONCLUSION: OPG mRNA is expressed in peripheral blood mononuclear cells and known to be increased by IL-1beta.

Telomerase Activity and Expression of MIB-1 and bcl-2 in Human Chorionic Villi from Early and Term Normal Pregnancy

Telomerase is an enzyme that maintains telomeres and prevents telomere shortening, and may be linked with cellular proliferation or the aging process. The purpose was to examine telomerase activity in human chorionic villi from early and term normal pregnancies, and to analyze the correlation of telomerase activity (TA) with MIB-1 & bcl-2. A total of 37 placentae were obtained from 16 early and 21 term pregnancies. TA was assayed by telomeric repeat amplification protocol, and immunohistochemical staining was performed for MIB-1 & bcl-2 expression. TA & MIB-1 expression were strong in early placenta, but bcl-2 was highly expressed in term placentae. Thirteen (81.25%) of 16 early placentae showed TA, but only 2 (9.52%) of 21 term placentae expressed TA (p<0.01). MIB-1 was observed in nuclei of cytotrophoblast, and the expression rate was 16.09% in early placentae and 2.87% in term placentae (p<0.01). bcl-2 was observed only in the cytoplasm of syncytiotrophoblast. Term placenta demonstrated stronger expression of bcl-2 compared to early placentae (p<0.05). These findings suggest that TA, MIB-1 & bcl-2 expression are critically regulated over the course of gestation: cytotrophoblast, main cells of early chorionic villi, may be a common source of telomerase and proliferative activity. The TA showed good correlation with cellular proliferative activity. Syncytiotrophoblast, may be a main source of bcl-2 expression which is stronger in the term placentae.

Telomerase Activity in Human Breast Tumors

BACKGROUND: Telomerase is a ribonucleoprotein enzyme that synthesizes telomeric DNA onto the ends of chromosomes, thereby preventing the replication-dependent shortening of those ends. Telomerase activity is detected in a wide range of cancers of various tissues, and its expression may be a critical step in tumor progression. Our objective was to determine if detection of telomerase activity may be an indicator for diagnosis of breast cancer and if any association exists between telomerase activity and prognostic factors of breast cancer. METHODS: Using a polymerase chain reaction-based telomerase activity assay, we examined telomerase activity in 30 breast cancer specimens (2 ductal carcinoma in situ, 28 invasive ductal carcinoma), 25 benign lesions (14 fibroadenomas, 11 fibrocystic diseases), and 24 normal breast tissues (13 adjacent to malignancy, 11 adjacent to benign lesion). RESULTS: Among surgically resected samples, telomerase activity was detected in 23 (77%) of 30 breast cancers. While telomerase activity was not detected in any of the 11 specimens of fibrocystic disease and the 11 normal tissues adjacent to benign lesion, surprisingly low levels of telomerase activity were detected in 5 (36%) of the 14 fibroadenomas and 1 (7%) of the 13 normal tissues adjacent to malignancy. There was no significant difference in expression of telomerase among prognostic factors of breast cancer. CONCLUSIONS: In summary, telomerase activity may be useful in the diagnosis of breast cancer. We found no correlation between telomerase activity and stage, tumor size, or LN status. Mechanisms of telomerase expression are still under investigation; therefore, the significance of telomerase expression in malignant tumors and their progression remains to be determined.

A Case of the McCune: Albright Syndrome Associated with Activating Mutations of Stimulatory G Protein / 대한내분비학회지

McCune-Albright syndrome (MAS) is a sporadic disease classically including polyostotic fibrous dysplasia, cafe -au-lait spots, sexual precocity, and other hyperfunctional endocrinopathies. Recent investigations suggest an etiological role for activating embryonic somatic missense mutations in the gene for the a subunit of Gs (Gsa), the G protein that stimulates adenylyl cyclase. DNA from bone, ovary, and blood was analyzed by using polymerase chain reaction and sequenced. A embryological somatic mutation of Gsa gene encoding substitution of a Cys for Arg at amino acid 201 from cells of dysplastic bone and ovary was observed, and the distribution of mutant gene reveals mosaic pattern. We report a case of McCune-Albright syndrome with an activating mutation at codon 201 of Gsa subunit on ovary and bone tissue that was experienced recently.

Telomerase Activity in Human Breast Tumors / 한국유방암학회지

Telomerase is a ribonucleoprotein enzyme that synthesizes telomeric DNA onto the ends of chromosomes. thereby preventing the replication-dependent shortening of these ends. Telomerase activity is detected in a wide range of cancers of various tissues, and its expression may be a critical step in tumor progression. Our objective was to determine if detection of telomerase activity may be an indicator for diagnosis of breast cancer and any association between telomerase activity and prognostic factors of breast cancer. Using a polymerase chain reaction-based telomerase activity assay, we examined telomerase activity in 30 breast cancer specimens (2 ductal carcinoma in situ, 28 invasive ductal carcinoma), 25 benign lesions (14 fibroadenomas, 11 fibrocystic diseases) and 24 normal breast tissues (13 adjacent to malignancy, 11 adjacent to benign lesion). Among surgically resected samples, telomerase activity was detected in 23 (77%) of 30 breast cancers. While telomerase activity was not detected in any of 11 specimens of fibrocystic disease and 11 adjacent normal tissues to benign lesion, surprisingly low levels of telomerase activity were detected in 5 (36%) of 14 fiboadenomas and 1 (7%) of 13 adjacent normal tissues to malignancy. There was no significant difference in expression of telomerase among prognostic factors of breast cancer. In summary, telomerase activity in breast cancer may be useful in diagnosis of breast cancer. We found no correlation between telomerase activity and stage, tumor size or LN status. Mechanisms of telomerase expression are still under investigation; therefore, the significance of telomerase expression in malignant tumors and their progression remains to be determined.

Correlation of Heregulin mRNA and Her-2/neu Protein Expression with Node Metastasis and DNA Ploidy Pattern in Human Invasive Breast Carcinoma

The Her-2/neu protooncogene encodes a transmembrane tyrosine kinase that is structurally homologous to the receptor for epidermal growth factor. Its amplification and overexpression are associated with poor prognosis in breast cancer patients. Neu differentiation factor is a ligand for Her-2/neu protooncogene and was detected in ras-transformed rat fibroblasts. Heregulin (human homologue of neu differentiation factor) is a 44-kilodalton glycoprotein that stimulates tyrosine phosphorylation and induces growth arrest or stimulation and differentiation in human breast cancer cell lines. In this study we examined the expression of heregulin mRNA by nested reverse transcription (RT) PCR with fresh tissue, Her-2/neu protein, ICAM-1 and steroid receptors by immunohistochemistry, and DNA ploidy pattern by flow cytometry with paraffin-embedded tissue in invasive breast carcinoma. We compared the data with nodal status, lymphovascular invasion, steroid receptor status and DNA ploidy pattern. For RT-PCR to heregulin mRNA, 38 cases of fresh breast cancer tissue were obtained. Total 68 cases of invasive breast carcinoma tissue were fixed in formalin, which were used for routine histology, immunohistochemistry and flow cytometry. The results are as follows; 1) Heregulin mRNA was expressed in 86.1% of patients with invasive breast carcinoma and 100% of patients with benign breast lesion using nested RT-PCR analysis. 2) Her-2/neu protein was overexpressed in 50.0% of tumors using immunohistochemistry. The expression of Her-2/neu protein was significantly correlated with high counts of lymph nodes with metastasis (p<0.05), and high nuclear grade (p<0.05). 3) Her-2/neu protein overexpression was significantly correlated with a high DNA index(p<0.05). All of the tumors showing Her-2/neu protein overexpression and no heregulin mRNA expression revealed near tetraploid DNA content. However, both Her-2/neu overexpression and heregulin mRNA expressing tumors revealed near tetraploidy in 38.9% and diploidy in 50.0%. Based on these results, heregulin mRNA expression rate was 86.1% in human invasive breast carcinoma. Her-2/neu protein overexpression is associated with high positive lymph node number and DNA index. Statistically significant reverse correlation with lymph node metastasis is not present.

A Study for IL-6, IL-13 and TIMP-3 Expressions of Placenta, Fetus and Endometrium in Pregnant Mice after Treatment with Lipopolysaccharide

We examined C3H pregnant mice at 15 days (70% gestation) after treatment of lipopolysaccaride (LPS) to observe the changes of IL-6 concentration in maternal serum and amniotic fluid and expression of IL-6, IL-13 & TIMP-3 in placenta, fetus and endometrium, and to investigate the correlation among IL-6, IL-13 and TIMP-3. The results were as follows: 1) IL-6 in serum and amniotic fluid after treatment of LPS was significantly elevated; peaked at 1, 2, 4, 5 hours and decreased to control level at 8 hours (P<0.05). IL-6 in placental disc, chorioamnionic membrane, fetus, decidua and endometrial epithelium was overexpressed significantly at 1, 2, 4 hours after treatment of LPS (P<0.05). IL-6 overexpression was more significantly increased in maternal tissue than fetal tissue (P<0.05). 2) Increased concentration of amniotic fluid IL-6 was equally originated from transplacental crossage of maternal serum IL-6, and direct local production of IL-6 from placenta, fetus and endometrium (P<0.05). 3) IL-13 in placental disc, chorioamnionic membrane, fetus, decidua and endometrial epithelium was overexpressed after treatment of LPS, but not significant statistically. 4) TIMP-3 was overexpressed in placental disc, chorioamnionic membrane, fetus and decidua. TIMP-3 overexpression was more significant in placental disc than other tissues (P<0.05). 5) Overexpressions in IL-13 and IL-6 revealed direct proportional correlation coefficient (Spearman correlation coefficient, 0.5212 ; P<0.05). IL-6 expression was a head of overexpression of TIMP-3, but not significant. In conclusion, all of IL-6, IL-13 and TIMP-3 relate with inflammatory response, especially IL-6 in maternal serum, amniotic fluid and tissue of placenta, fetus and endometrium was so sensitive that it can be an indicator for antenatal diagnosis of chorioamnonitis, and amniotic fluid IL-6 is equally originated from maternal serum and from tissue of placenta, fetus and endometrium. IL-13 and TIMP-3 may have parallel correlation to the IL-6 in fetal and maternal tissue after treatment of LPS.

Non-association of Pvull and Xval Estrogen receptor Genotypes with Bone Mineral Density and Bone Markers in Korean Premenopausal Women / 대한내분비학회지

BACKGROUND: Bone mineral density (BMD) is under strong genetic control. A recently reported case of severe estrogen resistance caused by a germ-line mutation at the estrogen receptor gene locus suggests the possibility that other variants of the estrogen receptor (ER) gene could be responsible for the heritable components of bone density. METHODS: Two restriction fragment length polymorphisms (RFLPs) at the ER gene locus, represented as PvuII and XbaI, and their relationship to bone mineral density (BMD) and bone turnover markers were examined in 95 healthy premenopausal women. Their mean age was 29 +-6.9 years (mean+-SD). RESULTS: The distribution of the PvuII and XbaI RFLPs was as follows: PP 20 (21.1%), Pp 40 (42.1%), pp 35 (36.8%), and XX 5 (5.3%), Xx 33 (34.7%), xx 57 (60.0%) (capital letters signify the absence of, and lower case letters signify the presence of the restriction site of each RFLP). There was no significant relation between ER genotypes and BMD measured at several sites such as lumbar spine (L2-4), distal forearm, and femoral neck. Also no significant genotypic differences were found in the several biochemical markers and sex hormone status. CONCLUSION: These data indicate that these polymorphisms are not predietive of bone turnover nor BMD in a sample of healthy Korean premenopausal women.

CD5 mRNA Overexpression in Lymphocytes of Allergic Patients / 대한면역학회지

PURPOSE: The CD5 molecules are pan-T cell antigens and are found on a minor subpopulation of B cells. CD5 antigens are involed in an intracellular signal transduction as well as in an intercellular signal transduction between CDS+ T cell/CD72+ B cell by CD5/CD72 interaction. CD5 antigens are known to be participated in classic immune reactions and in this study CDS mRNA expressions by lymphocytes were examined in allergic patients controls, acute febrile infectious disease controls and normal controls to elucidate the possibility of CDS involvement in allergic immune reactions. METHODS: Fifteen allergic patients, ten patients of acute febrile infectious disease patients and ten normal controls were studied. Venous blood was drawn and mononuclear cells were separated. T cells and B cells were separated using immunomagnetic beads. Total RNA was extracted and RT-PCR (reverse transcriptase - polymerase chain reaction) was done to detect CDS antigen mRNA expression. RESULTS: 1) CDS mRNA overexpressions were detected in allergic patient controls as compared to that in acute febrile infectious controls. CDS mRNA was not detected in normal controls. Semiquantitative CD5 mRNA expressions were measured as relative expressions of CD5 to GAPDH. Relative quantities of CD5 mRNA expressions were 90.656.24% in allergic patient controls and 23.76+3.58% in acute febrile infectious patients. CONCLUSIONS: CDS mRNA overexpression is a characteristic phenomenon in allergic immune reactions. From these result, CD5/CD72 pathway might be the preference immune mechanism in allergic immune reaction and the further study for the exact mechanism of CDS involvement in allergic immune reactions may be necessary

Effect of Dexamethasone and Deflazacort on the Function and Gene Expression of the Primary Cultured Human Osteoblast-Like Cells / 대한내분비학회지

Background: Chronic use of glucocorticoid is known to result in osteoporosis. Deflazacort (DFZ), a synthetic glucocorticoid, has been reported to have bone sparing properties in vivo eompared to dexamethasone(DEX). Not only the direct effect of DFZ on human osteoblast but the mechanism by which the drug spares bone remains unclear. This study, therefore, is aimed to investigate the direct effect of DFZ on the proliferation and differentiation of human osteoblast as well as on the gene expression of osteocalcin and osteoblast as well as on the gene expression of osteocalcin and growth factor produced in osteoblast. Methods: Human osteoblast-like cells were cultured from a piece of the tibia removed during selective orthopedic surgery for patients without metabolic bone diseases. The morphological iden- tification of osteoblast-like cell was performed under the light microscope after alkaline phosphatase staining. Cell proliferation rate was determined by [3H] thymidine incorporation into DNA. Cell differentiation was determined by alkaline phophatase activity. mRNA expression was quanti- tatively measured by the competitive reverse transcription-polymerase ehain reaction(RT-PCR). Results: The cultured cells demonstrated 1,25-dihydroxyvitamin D3-induced increases in alkaline phophatase activity and osteocalcin mRNA expression which are the properties of osteoblast. Twenty six percent of the cultured cells were identified as osteoblast-like cells by alkaline phophatase staining. After 24hr incubation with DEX or DFZ, the [3H) thymidine incorporation was significantly inhibited by 100nM DEX or DFL Alkaine phophatase activity was significantly increased by 100nM DEX. Osteocalcin mRNA was significantly decreased by both glueocorticoids. While DEX significantly suppressed expression of asteocalcin mRNA at 10nM and 100nM, DFZ did so only at 100nM. IGF-I mRNA was significantly decreased by 100nM DEX. Conclusion: These results suggest that the inhibitory effect of DFZ on the cell proliferation and protein synthesis is less than that of DEX, which might be responsible for the bone sparing effect of DFZ in vivo.

Rapid determination of fetal Y-chromosome with polymerase chain reaction / 대한산부인과학회잡지

No abstract available.

The Pharmacokinetics of Lidocaine after Tracheal Administration during General Anesthesia / 대한마취과학회지

Spinal Anesthesia employing 0.5% plain bupivacaine was administered to 40 patients scheduled for lower limb or perineum sThe plasma concentrations and pharmacokinetic parameters of lidocaine were studied in 4 patients under general anesthesia(halothane, or enflurane-N20-O2) following the introduction of an 1% lidocaine endotracheal spray(1.5mg/kg) through an epidural catheter. Poak plasma lidocaine levels were reached in 5 to 15 minutes and were within the nontoxic range. The pharmacokinetics of lidocaine in these patients can be described by a two-compartment model with a rapid alpha distribution(T 1/2 alpha 8.66+/-2.24 min.), and an extensive apparent volume of idstribution(1.32+/-0.46 1/kg) similar to that observed in normal subjects. The half-life of absorption was 3.65+/-1.21 minutes. However, the elimination half-life(T 1/2 beta 173.25+/-32.41 min.) was prolonged and the total plasma clearance( 6.15+/-3.25 ml/min/kg) was decreased. This potent inhalation anesthetic agent may reduce the hepatic blood flow and would be expected to reduce the plasma clearance of lidocaine by reducing the delivery of plasma lidocaine. This study suggests that tracheal administration of lidocaine will produce effective plasma lidocaine levels in many clinical situations as will intravenous administration.