Expression of Heat Shock Protein 60 kDa Is Upregulated in Cervical Cancer

PURPOSE: Cervical cancer caused by the human papilloma virus (HPV) continues to be the cause of yearly death among women. However, it is a curable disease when diagnosed at an early stage. Recently, several researches have reported that heat shock protein (HSP) 60, a chaperone protein of molecular weight of 60 kDa, is involved in carcinogenesis and apoptosis. In order to evaluate the prognostic significance of HSP60 in cervical cancer, we examined differences in the HSP60 expression between cervical cancer and normal tissues in women. MATERIALS AND METHODS: Tissue samples were collected from 20 cervical cancer patients and 20 normal controls. HSP60 expression of cervical cancer and normal tissues were verified by the 2D gel proteomics, semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot analyses. RESULTS: In 2D proteomic analysis, an increase of HSP60 expression was detected in cervical cancer tissues and confirmed by Western blot analysis (p < 0.05). However, messenger RNA (mRNA) levels of HSP60 did not display any significant differences between cervical cancer and normal tissues. CONCLUSION: These results suggest that HSP60 may be involved in the development of cervical cancer and have profound biological and prognostic significance.

Rarity of TLR4 Asp299Gly and Thr399Ile Polymorphisms in the Korean Population

PURPOSE: Activation of the innate immune system and chronic low-grade inflammation are thought to be involved in the pathogenesis of atherosclerosis and also thought to be associated with type 2 diabetes and its complications. As a receptor for bacterial lipopolysaccharide and heat-shock proteins, Toll-like receptor 4 (TLR4) is one of the central regulators of the immune response. Recent studies have reported an association between TLR4 polymorphisms and diabetes and its complications in Caucasian populations. MATERIALS AND METHODS: In this study, we analyzed the association between TLR4 gene polymorphisms in patients with features of type 2 diabetes and healthy controls in Korea. Two polymorphisms of the TLR4 gene (Asp299Gly and Thr399Ile) were examined in 225 diabetic patients and 153 healthy controls using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and single-strand conformation polymorphism (SSCP). RESULTS: No Asp299Gly or Thr399Ile mutations were detected in any of the 378 subjects. Seven subjects from each group who had slightly different SSCP patterns were selected for sequencing, but we found no TLR4 polymorphisms on Exon3. The Asp299Gly and Thr399Ile TLR4 gene polymorphisms were absent in both groups, which was similar to the results for Japanese and Chinese Han subjects. CONCLUSION: Our data and other Asian data suggest that a racial difference can be found in the frequency of the TLR4 polymorphism.

Heat Shock Protein 60 and 27 expressions on the trophoblast of preterm birth / 대한산부인과학회지

OBJECTIVE: To compare the distribution and expression of hsp60 and hsp27 in placental trophoblast between preterm and term placenta and to observe hsp immune response in relation to the pathogenesis of preterm birth. METHODS: 22 cases of preterm trophoblast, between 24 weeks and 36 weeks gestation, which were developed spontaneous onset or less than 24 hours after rupture of membrane were obtained. And aged-matched, 22 cases of normal term trophoblast, as control were also obtained after informed consent from each patient. The protein extraction form trophoblast was stained by immunohistochemical methods and was measured by the assay of Western blots. And the density of band using Image-writer were taken and statistical assay were performed as significance <0.05. RESULTS: The expressions of hsp60 and hsp27 in trophoblast of preterm and term placenta were identified by immunohistochemical staining method. The hsp60 had significantly higher expression in trophoblast of preterm birth than in that of term birth (P<0.001) and the hsp27 also had significantly higher expression in trophoblast of preterm birth than in that of term birth (P<0.02) CONCLUSION: The higher expression of hsp60 and hsp27 in trophoblast of preterm birth might be suggested the development of immune response to occur preterm labor Further study are necessary to determine the exact actions of hsp60 and27 in trophoblast and to understand the immune mechanism of preterm birth.

Glutathione S-Transferase P1 Polymorphism in Patients with Allergic Rhinitis / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Genetic factors play a role in the etiology of allergic rhinitis. The glutathione S-transferase (GSTP1) is one of the detoxifying enzymes for oxidative stress. The purpose of this study is to examine Polymorphisms of whether there is an association between some alleles of GSTP1 genes and allergic rhinitis. SUBJECTS AND METHOD: Patients with allergic rhinitis were selected on the basis of the following criteria: 1) watery rhinorrhea, sneezing, nasal obstruction and/or itching for longer 3months and 2) positive reaction at the allergic skin prick test for DP, DF allergen and 3) positive reaction at specific IgE RAST for DP, DF allergen. GSTP1 gene polymorphisms in exon5 (Ile105Val) were analyzed by polymerase chain reaction (PCR) with restriction fragment length polymorphisms (RFLP) in 149 patients with allergic rhinitis and 156 healthy control subjects. RESULTS: In allergic rhinitis, Ile105/Ile105 were 106 cases (71.1%), Ile105/Val105 were 42 cases (28.2%), Val105/Val105 were 1 case (0.7%) and in normal controls, Ile105/Ile105 were 100 cases (64.1%), Ile105/Val105 were 45 cases (28.8%), Val105/Val105 were 11 cases (7.1%)(p=0.004). CONCLUSION: Genetic polymorphism of Val105/Val105 in GSTP1 may be protective genotypes in allergic rhinitis.

C-met and E-cadherin Expression in Advanced Gastric Cancer

PURPOSE: In the current study, the relation between the clinicopathological parameters and levels of the amplification of the c-met and E-cadherin genes were investigated in patients with an advanced gastric carcinoma. METHODS: The levels of amplification of the c-met and E-cadherin genes in 44 advanced gastric carcinoma patients were retrospectively investigated using RT-PCR. The relationships between the levels of amplification of these genes and the clinicopathological parameters were evaluated using univariate and multivariate analyses. RESULTS: Seventeen (38.6%) and 13 (29.5%) of the 44 advanced gastric carcinoma patients were evaluated as having amplification of the c-met gene and down-regulation of the E-cadherin gene, respectivly. The amplification of c- met gene was significantly correlated with serosal invasion, lymph node metastasis and neural invasion, whereas the down-regulation of the E-cadherin gene was significantly correlated with the diffuse type of gastric carcinoma by Lauren's calssification, and neural invasion. CONCLUSION: The levels of the c-met and E-cadherin gene amplifications may be a powerful aids in evaluating the metastatic potential and prognosis in patients with advanced gastric cancer.

Detection of Chlamydia pneumoniae in Korean Patients with Abdominal Aortic Aneurysm or Peripheral Arterial Occlusive Disease

PURPOSE: Chlamydia pneumoniae (CP) infection seems to be related to atherosclerotic diseases. A prospective sero- epidemiologic study was performed to analyze the relationship between CP infection and peripheral vascular disease in Korean patients. The aims of this study were to find the prevalences of CP antibody in the serum and CP antigens in the vascular tissues, and to analyze the differences between several disease groups. METHODS: Our subjects included a total of 61 patients (76 vascular tissues) who had undergone operative procedures for peripheral vascular diseases. They were classified into 3 groups: Group 1; 14 abdominal aortic aneurysm, Group 2; 15 atherosclerosis obliterans, and Group 3; 32 varicose vein cases. The CP antibody titers were determined using the microimmunofluorescence test (MIF) and the CP antigen in the vascular tissues with a semi-nested polymerase chain reaction (PCR) and an in situ hybridization technique (ISH). RESULTS: The prevalences of chronic or past CP infection from the MIF (IgG antibody titer > or = 1: 32) in Groups 1, 2 and 3 were 78.6, 73.3, and 68.8% respectively, but with statistically significant differences. The prevalences of PCR- positive tissues in Groups 1, 2 and 3 were 21.4, 6.7, and 0% respectively. There was no CP DNA detected in the venous tissue. CP DNA was detected more frequently in aneurysmal disease than atherosclerosis obliterans, but this was not statistically significant(p=0.265). In comparison with the varicose veins, aortic aneurysms showed a significantly higher PCR positivity ratio (p=0.002), and a similar result was seen with ISH. There was no relationship between CP antigen positivity and the known risk factors for atherosclerosis. CONCLUSION: A high prevalence of CP antibodies was observed in the serum of Korean patients with vascular disease, which matched that in western populations. CP DNA was also detected in atherosclerotic tissues, which was especially high in aneurysmal disease, implying a possible causative role of CP infection in the pathogenesis of the atherosclerotic disease. This is the first report on the prevalence of CP in vascular tissues in Korean population.

Effect of Long-Term Low-Dose Macrolide Therapy in Children with Chronic Rhinosinusitis

Backgrounds and Objectives: We studied the effect of long-term low-dose macrolide therapy on the level of IL-8, IL-1 beta, and TNF-alpha in the nasal secretions get from children with chronic rhinosinusitis before and after medication, and investigated the association between the changes in the chemical mediator levels and the clinical outcome before and after macrolide treatment. MATERIALS AND METHODS: The nasal lavage was obtained from 10 patients with nonallergic chronic rhinosinusitis and also from 10 healthy children. Nasal lavage was obtained before and 1 month after full dose (8 mg/kg ) macrolide administration, and then second lavage was obtained after half dose (4 mg/kg ) administration for 2 month. The level of IL-8, IL-1 beta and TNF-alpha in nasal lavage were measured by using ELISA kit. The symptoms were also scored by visual analogue scale before and after treatment. RESULTS: The IL-8 concentration was decreased from 317.4 pg/ml to 227.1 pg/ml at 12 weeks after this treatment (p<0.05). The level of IL-1beta was decreased from 412.5 pg/ml to 41.5 pg/ml (p<0.05), and TNF-alpha was also decreased from 49.8 pg/ml to 3.9 pg/ml (p<0.05). The symptoms of most patients with macrolide treatment were improved at 3 months after treatment. CONCLUSION: Macrolide decreased the concentration of inflammatory mediators in nasal discharge, such as IL-8, IL-1beta and TNF-alpha and this anti-inflammatory effect of macrolide could explain the way of improvement with subclinical dosage of drug.

Clinical Usefulness of Plasma Activities of Gelatinase (Matrix Metalloproteinase-2 and 9) in Chronic Liver Disease / 대한간학회지

BACKGROUND/AIMS: Gelatinase (matrix metalloproteinase (MMP) -2 and 9) has an important role in the pathogenesis of liver cirrhosis (LC) and hepatocellular carcinoma (HCC). In this study, we evaluated the relationship of gelatinase to chronic liver disease. METHODS: Four groups of subjects were examined; healthy control (10 cases), chronic hepatitis (18 cases), LC (15 cases), and HCC (28 cases). The plasma of each subject was obtained, and the equal quantification of plasma protein was done. The plasma activities of MMP-2 and 9 were measured by zymography. RESULTS: The activities of plasma MMP-2 in patients with LC were significantly higher than those in controls (p=0.009) and in patients with chronic hepatitis (p=0.011), but not different from those in patients with HCC. The activities of plasma MMP-9 in patients with LC were significantly higher than those in controls, but not different from those in patients with chronic hepatitis or HCC. In patients with LC (regardless of having HCC), the activities of MMP-2 correlated with total bilirubin (r=0.323, p=0.048) and Child-Pugh score (r=0.414, p=0.012). The activities of MMP-2 and 9 were higher in patients with LC (regardless of having HCC) caused by alcohol than caused by HBV (p=0.009 and 0.002 for each one). CONCLUSIONS: The plasma activity of MMP-2 may be a useful marker for the diagnosis and determination of the severity of LC. The plasma activity of MMP-9 was not useful for HCC, but may be a marker for alcoholic LC. Further study is needed to determine why the plasma activity of gelatinase was higher in patients with LC caused by alcohol than by HBV.

Polymorphism in Codons 10 and 25 of the Transforming Growth Factor-beta1 Gene in Korean Population and in Patients with Liver Cirrhosis and Hepatocellular Carcinoma / 대한소화기학회지

BACKGROUND/AIMS: The genetic polymorphism of transforming growth factor-beta1 (TGF-beta1) at codons 10 and 25 which influences the production of TGF-beta1 is related to fibrogenesis in the lung and liver. We evaluated the genetic polymorphism at codons 10 and 25 in controls and in patients with liver cirrhosis (LC) and hepatocellular carcinoma (HCC). METHODS: Blood samples were collected from controls (n=35), patients with LC (n=64), and HCC (n=49). Genomic DNA was isolated and polymerase chain reaction (PCR) was done for a segment including codons 10 and 25. The results of direct sequencing for PCR products were compared between the controls and the patients. RESULTS: There was no genetic polymorphism at codon 25 and three types of genetic polymorphism at codon 10. The leucine homozygous genotype (CTG/CTG) at codon 10 was more common in patients with LC than the controls (p=0.01) and especially in patients with LC caused by HBV (p=0.004). The polymorphism at codons 10 in patients with HCC was similar to the controls. However, leucine homozygous genotype was more common in patients with HCC of uninodular morphology than those of massive morphology (p=0.007). CONCLUSIONS: The genetic polymorphism of TGF-beta1 at codon 10 might be associated with LC and morphology of HCC. The potential usefulness of TGF-beta1 genotyping needs further studies in large scale.

Gene Expression Profiles in Cervical Cancer with Radiation Therapy Alone and Chemo-radiation Therapy / 대한방사선종양학회지

PURPOSE: To analyze the gene expression profiles of uterine cervical cancer, and its variation after radiation therapy, with or without concurrent chemotherapy, using a cDNA microarray. MATERIALS AND METHODS: Sixteen patients, 8 with squamous cell carcinomas of the uterine cervix, who were treated with radiation alone, and the other 8 treated with concurrent chemo-radiation, were included in the study. Before the starting of the treatment, tumor biopsies were carried out, and the second time biopsies were performed after a radiation dose of 16.2~27 Gy. Three normal cervix tissues were used as a control group. The microarray experiments were performed with 5 groups of the total RNAs extracted individually and then admixed as control, pre-radiation therapy alone, during-radiation therapy alone, pre-chemoradiation therapy, and during-chemoradiation therapy. The 33P-labeled cDNAs were synthesized from the total RNAs of each group, by reverse transcription, and then they were hybridized to the cDNA microarray membrane. The gene expression of each microarrays was captured by the intensity of each spot produced by the radioactive isotopes. The pixels per spot were counted with an Arrayguage(R), and were exported to Microsoft Excel(R). The data were normalized by the Z transformation, and the comparisons were performed on the Z-ratio values calculated. RESULTS: The expressions of 15 genes, including integrin linked kinase (ILK), CDC28 protein kinase 2, Spry 2, and ERK 3, were increased with the Z-ratio values of over 2.0 for the cervix cancer tissues compared to those for the normal controls. Those genes were involved in cell growth and proliferation, cell cycle control, or signal transduction. The expressions of the other 6 genes, including G protein coupled receptor kinase 6, were decreased with the Z-ratio values of below -2.0. After the radiation therapy, most of the genes, with a previously increase expressions, represented the decreased expression profiles, and the genes, with the Z-ratio values of over 2.0, were cyclic nucleotide gated channel and 3 Expressed sequence tags (EST). In the concurrent chemo-radiation group, the genes involved in cell growth and proliferation, cell cycle control, and signal transduction were shown to have increased expressions compared to the radiation therapy alone group. The expressions of genes involved in angiogenesis (angiopoietin-2), immune reactions (formyl peptide receptor-like 1), and DNA repair (cAMP phosphodiesterase) were increased, however, the expression of gene involved in apoptosis (death associated protein kinase) was decreased. CONCLUSION: The different kinds of genes involved in the development and progression of cervical cancer were identified with the cDNA microarray, and the proposed theory is that the proliferation signal starts with ILK, and is amplified with Spry 2 and MAPK signaling, and the cellular mitoses are increased with the increased expression of Cdc 2 and cell division kinases. After the radiation therapy, the expression profiles demonstrated the evidence of the decreased cancer cell proliferation. There was no significant difference in the morphological findings of cell death between the radiation therapy alone and the chemo-radiation groups in the second time biopsy specimen, however, the gene expression profiles were markedly different, and the mechanism at the molecular level needs further study.

Expression of Collectin in the Human Nasal Mucosa / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Collectins (surfactant protein A and D) are proteins with collagen tails and globular lectin domains that appear to play an important role in the first line of host defense in mammalians. However, it is not known if collectins are also present in human nasal mucosa. The purpose of this study was to investigate the expression of collectin proteins in human nasal mucosa and to compare the expressions of SP-A and D mRNA in the normal nasal mucosa and in chronic inflammatory nasal diseases. MATERIALS AND METHOD: Ten chronic rhinosinusitis patients were recruited and ten normal nasal mucosae were used as normal controls. Reverse transcriptase-polymerase chain reaction was performed to detect SP-A and SP-D mRNA. The level of collectin and GAPDH (glyceraldehyde-3-phosphate dehydrogenase) transcripts were semi-quantified with the desitometry. We have characterized the cellular localization of SP-A and SP-D protein using immunohistochemistry. RESULTS: SP-A2/GAPDH mRNA ratio in chronic rhinitis nasal mucosa is greater compared with that in normal nasal mucosa (p<0.05). SP-A protein was expressed in the nasal epithelium and in the epithelial cells of the submucosal glands. SP-D mRNA and protein were not expressed in the nasal mucosa. CONCLUSION: These data provide the first evidence of the presence of collectins in the human nasal mucosa. These results suggested that up-regulation of collectin in chronic rhinosinusitis may be a protective response for the nasal mucosa.

The Functional and Genetic Defects of IFN-gamma Receptor in the Patients with Tuberculosis / 결핵

BACKGROUND: IFN-gamma plays an important role in the host response to a mycobacterial infection. A complete IFN-gamma receptor 1 deficiency is a life-threatening condition because it renders patients highly susceptible t o a mycobacterial infection. Several mutations in the IFN-gamma receptor and STAT1 gene have been identified in the rare mycobacterial infections. These mutations have partial function of the IFN-gamma receptor and similar pathologic features to clinical tuberculosis. METHODS: The function of the IFN-gamma receptor was evluated in the patients with clinical tuerculosis. In addition, the DNA coding sequence of the IFNgR1 and STAT1 gene was also analyzed in disseminated tuberculosis patients who might have a defective IFN-gamma receptor. RESULTS: The cell surface expression levels of HLA-DR and CD64 in the PMBC after being stimulation with IFN-gamma (100Imicro/ml, 1000Imicro/ml) were increased in both controls and patients. However, the rate of increase in both groups was similar. The production of TNF-alpha in the response to stimulation with LPS was higher in the both groups (850.7+/-687.8 vs. 836.7+/-564.3 pg/ml). Pretreatment with IFN-gamma prior to LPS stimulation resulted infurther increase in TNF-alpha production in the both groups was similar. The known mutations in the IFNgR1 and STAT1 coding sequences were not found in the genomic DNA of patients wit disseminated tuberculosis. CONCLUSION: The functional and genetic defects of the IFN-gamma receptor were not identified in clinical tuberculosis. This suggests the defective IFN-gamma receptor that predispoe patiens to a BCG or NTM infection can not alone account for the cases of clinical tuberculosis.

The Expression of Gastrin and Gastrin/ CCK-B Receptor mRNA in Cancer and Normal Tissue of Large Intestine

PURPOSE: Gastrin, a peptide hormone produced by the G cells of the gastric antrum, plays a major role in regulating acid secretion in the stomach, and acts as a trophic factor in the gastrointestinal tract. The relationship between gastrin and the development of colorectal cancer remains controversial. To study its possible role in development or proliferation of colorectal cancer, we evaluated the expression of gastrin and gastrin/CCK-B receptor mRNA in cancer and normal tissue from colorectal cancer patients. We also reviewed clinical records to evaluate the correlations between gastrin receptor expression and clinical characteristics of colorectal cancer. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) was used to evaluate mRNA expression for gastrin and gastrin/CCK-B receptor in 26 surgical specimens of colorectal cancer. RESULTS: The mRNA expression of gastrin was detected in 24 out of 26 cancer specimens and 9 out of 26 normal colon specimens (p0.05). There was no significant correlation between gastrin receptor expression and clinical characteristics of colorectal cancer. CONCLUSIONS: The gastrin gene products might be more important than gastrin/CCK-B receptor in development or proliferation of colorectal cancer, which supports the hypothesis that gastrin gene products play a role in proliferation of colorectal cancer as an autocrine factor.