摘要 :
We investigated the roles of interleukin-6 (IL-6) and parathyroid hormone-related peptide (PTHrP) in oral squamous cell carcinoma (OSCC)-induced osteoclast formation. Microarray analyses performed on 43 human OSCC specimens reveal...
展开
We investigated the roles of interleukin-6 (IL-6) and parathyroid hormone-related peptide (PTHrP) in oral squamous cell carcinoma (OSCC)-induced osteoclast formation. Microarray analyses performed on 43 human OSCC specimens revealed that many of the specimens overexpressed PTHrP mRNA, but a few overexpressed IL-6 mRNA. Immunohistochemical analysis revealed that IL-6 was expressed not only in cancer cells but also in fibroblasts and osteoclasts at the tumor-bone interface. Many of the IL-6-positive cells coexpressed vimentin. Conditioned medium (CM) derived from the culture of oral cancer cell lines (BHY, Ca9-22, HSC3, and HO1-u-1) stimulated Rankl expression in stromal cells and osteoclast formation. Antibodies against both human PTHrP and mouse IL-6 receptor suppressed Rankl in ST2 cells and osteoclast formation induced by CM from BHY and Ca9-22, although the inhibitory effects of IL6 antibody were greater than those of PTHrP antibody. CM derived from all of the OSCC cell lines effectively induced IL-6 expression in stromal cells, and the induction was partially blocked by anti-PTHrP antibody. Xenografts of HSC3 cells onto the periosteal region of the parietal bone in athymic mice presented histology and expression profiles of RANKL and IL-6 similar to those observed in bone-invasive human OSCC specimens. These results indicate that OSCC provides a suitable microenvironment for osteoclast formation not only by producing IL-6 and PTHrP but also by stimulating stromal cells to synthesize IL-6.
收起
摘要 :
The clustered regularly interspaced short palindromic repeat (CRISPR) systems composed of DNA direct repeats designated as CRISPRs and several CRISPR-associated (cas) genes, which are present in many prokaryotic genomes, make up a...
展开
The clustered regularly interspaced short palindromic repeat (CRISPR) systems composed of DNA direct repeats designated as CRISPRs and several CRISPR-associated (cas) genes, which are present in many prokaryotic genomes, make up a host defense system against invading foreign replicons such as phages. In order to investigate the altered expression profiles of the systems after phage infection using a model organism, Thermus thermophilus HB8, which has 12 CRISPR loci, genome-wide transcription profiling of the strain infected with lytic phage PhiYS40 was performed by DNA microarray analysis. Significant alteration of overall mRNA expression gradually increased during infection (i.e., from the eclipse period to the period of host cell lysis). Interestingly, the expression of most cAMP receptor protein (CRP)-regulated genes, including two CRISPR-associated (cas) operons, was most markedly up-regulated, especially around the beginning of host cell lysis, although up-regulation of the crp gene was not observed. The expression of the CRP-regulated genes was less up-regulated in a crp-deficient strain than in the wild type. Thus, it is suggested that cAMP is a signaling molecule that transmits information on phage infection to CRP to up-regulate these genes. On the other hand, the expression of several cas genes and that of CRISPRs were up-regulated independent of CRP, suggesting the involvement of unidentified regulatory factor(s) induced by phage infection. On analysis of the expression profile of the entire genome, we could speculate that upon phage infection, the signal was transmitted to the cells, with host response systems including CRISPR defense systems being activated, while the overall efficiencies of transcription, translation, and metabolism in the cells decreased. These findings will facilitate understanding of the host response mechanism following phage infection.
收起
摘要 :
In mammals, sex is determined by the presence or absence of the Y chromosome that bears a male-dominant sex-determining gene SRY, which switches the differentiation of gonads into male testes. The molecular signaling mechanism tur...
展开
In mammals, sex is determined by the presence or absence of the Y chromosome that bears a male-dominant sex-determining gene SRY, which switches the differentiation of gonads into male testes. The molecular signaling mechanism turning on the switch, however, has remained unclear for 18 years since the identification of the gene. Here, we describe how this gene emerged and started to work. From amino acid homology, we realized that SRY is a hybrid gene between a portion of the first exon of DiGeorge syndrome critical region gene 8 (DGCR8) and the high-mobility group (HMG) box of SRY box-3 (SOX3) gene. We identified the regulatory sequence in the SRY promotor region by searching for a common motif shared with DGCR8 mRNA. From the motif search between DGCR8 mRNA and the SRY upstream sequence, we found that the transcription factor CP2 (TFCP2) binding motif is present in both. TFCP2 overexpression did not show a significant increase of SRY mRNA expression, and TFCP2 suppression by RNA interference (RNAi) significantly reduced SRY mRNA expression. Furthermore, electrophoretic mobility shift assay (EMSA) demonstrated that TFCP2 acts as a regulator by directly binding to the SRY promoter. We conclude that SRY is a hybrid gene composed of two genes, DGCR8 and SOX3; and TFCP2 is an essential transcription factor for SRY expression regulation.
收起
摘要 :
OBJECTIVE: To develop a more rapid and accurate staining procedure for intraoperative cytology through an assessment of a rapid multiple immunocytochemical staining method using microwave irradiation. STUDY DESIGN: A preliminary t...
展开
OBJECTIVE: To develop a more rapid and accurate staining procedure for intraoperative cytology through an assessment of a rapid multiple immunocytochemical staining method using microwave irradiation. STUDY DESIGN: A preliminary test of a triple immunostaining method using microwave irradiation was performed to determine optimal incubating conditions for primary antibodies against MOC-31, BerEP4 and carcinoembryonic antigen. The test samples were adenocarcinoma cells obtained by washings from 10 resected colorectal cancer specimens. Adenocarcinoma cells in peritoneal washings diagnosed by cytologic examination from 8 colorectal cancer patients were retrospectively examined using the previously determined optimal incubating conditions. RESULTS: High rates of positive cells (83-87%) with intense immunoreactions were obtained using a rapid process with primary antibody concentrations that were 2 times higher than those used for standard incubation at room temperature (82-86%). The staining under these conditions was completed within 19 minutes. Adenocarcinoma cells in the peritoneal washings were also intensely stained under these conditions. CONCLUSION: Using our microwave method, the processing time was dramatically shortened, and intense and sensitive immunoreactions were obtained. The present method is useful for the rapid and accurate diagnosis of intraoperative cytology.
收起
摘要 :
In the present study, to investigate the effect of glucosamine, a component of glycosaminoglycans with a chondroprotective action, on articular cartilage in athletes, we looked at soccer players, who expose their joints to excessi...
展开
In the present study, to investigate the effect of glucosamine, a component of glycosaminoglycans with a chondroprotective action, on articular cartilage in athletes, we looked at soccer players, who expose their joints to excessive motion and loading, and compared the levels of biomarkers for type II collagen degradation (CTX-II) and type II collagen synthesis (CPII) between soccer players and non-athlete controls, and in soccer players before and after glucosamine-administration. CTX-II (P<0.01) and CPII (P=0.08) levels were substantially elevated in soccer players compared with those in controls, indicating that cartilage metabolism (type II collagen degradation and synthesis) is increased in soccer players. Of note, glucosamine administration (1.5 g and 3 g/day for 3 months) significantly decreased the CTX-II level (P<0.05); however, the effect disappeared after withdrawal of administration. In contrast, glucosamine administration did not essentially affect the increased level of CPII. Furthermore, cartilage damage was evaluated by using the ratio of type II collagen breakdown to synthesis (CTX-II/CPII). The ratio in soccer players was significantly higher than that in controls (P<0.05), suggesting that type II collagen degradation is relatively enhanced compared with type II collagen synthesis in soccer players than in control students. Of importance, the ratio was reduced by glucosamine administration but returned to the pre-administration level after withdrawal of administration. Together these observations suggest that glucosamine is expected to exert a chondroprotective action in athletes (soccer players) by preventing type II collagen degradation but maintaining type II collagen synthesis, although the effect is transient and disappears after withdrawal of administration.
收起
摘要 :
Transcription factors P53 and FOXO are both activated in response to stresses via protein-protein interactions, leading to events such as cell survival or apoptosis. To clarify the mechanisms that regulate FOXO activity, we analyz...
展开
Transcription factors P53 and FOXO are both activated in response to stresses via protein-protein interactions, leading to events such as cell survival or apoptosis. To clarify the mechanisms that regulate FOXO activity, we analyzed the intermolecular interaction of FOXO3a and P53. FOXO3a and P53 interacted in COS-7 cells, and transcriptional activity of FOXO3a was suppressed by P53, but P53 was not affected by FOXO3a. RT-PCR revealed that expression of the endogenous apoptosis-inducible genes Bim and Bcl6 was decreased markedly by co-expression of P53 with them, but expression of p27 and CyclinG2 was not. In addition, treatment with 500 microM H2O2 for 30 min to 1h to mimic oxidative stress promoted protein binding. Serum deprivation and drug treatment also affected the binding of FOXO3a and P53. These findings suggest that FOXO3a controls cellular function by changing its molecular interaction with P53 to mediate transcription factor activity under stress stimuli.
收起
摘要 :
We have investigated the effects of differential aspirin doses on atherogenesis. Aspirin was given to homozygous, apoE(-/-) and LDLR(-/-) double deficient mice for 12 weeks. The development of arteriosclerosis was determined morph...
展开
We have investigated the effects of differential aspirin doses on atherogenesis. Aspirin was given to homozygous, apoE(-/-) and LDLR(-/-) double deficient mice for 12 weeks. The development of arteriosclerosis was determined morphologically by image analysis and endothelial cell function was assessed by measurement of peripheral nitric oxide (NO). METHODS: ApoE(-/-) LDLR(-/-)double knockout mice were bred and maintained with a high fat diet containing aspirin (4 and 40 mg/kg B.W. /day) for twelve weeks. The development of arteriosclerosis was monitored by estimating the total area of atherosclerotic lesions in the entire aorta. Acetylcholine-induced NO release was measured in vivo using electrochemical sensors. The expression of eNOS on the endothelial surface was determined by immuno-staining. Plasma prostaglandin F1alpha (PGF(1 alpha)), serum thromboxian B(2) (TXB(2)) and total cholesterol were measured using enzymatic assay. Bleeding time was measured by tail cut method. RESULTS: Arteriosclerosis in the 4 mg/kg/day aspirin group was decreased significantly compared with the placebo group, but not in the 40 mg/kg/day aspirin group. Acetylcholine-induced NO release was significantly depressed in the 40 mg/kg/day aspirin group. Immunochemical analysis with anti-eNOS antibody supported these findings. In the 4 mg/kg/day aspirin group, the severe suppression of PGI(2) production was not confirmed in spite of decreasing TXB(2) production, but not in the 40 mg/kg/day aspirin group. CONCLUSION: Our results suggest that endothelial dysfunction with low dose aspirin improved, reduced progression of atherosclerosis in apoE(-/-) and LDLR(-/-) double deficient mice and provides a pathophysiological basis for the beneficial effects of aspirin in atherosclerosis, and low doses appeared to be more efficient than high doses.
收起
摘要 :
Kaposi's sarcoma-associated herpesvirus (KSHV) is related causally to Kaposi's sarcoma, primary effusion lymphoma, and a subset of cases of multicentric Castleman's disease. As the numbers of acquired immunodeficiency syndrome (AI...
展开
Kaposi's sarcoma-associated herpesvirus (KSHV) is related causally to Kaposi's sarcoma, primary effusion lymphoma, and a subset of cases of multicentric Castleman's disease. As the numbers of acquired immunodeficiency syndrome (AIDS) patients have increased, KSHV-associated diseases have also increased in Japan. Sporadic cases of classic Kaposi's sarcoma have also been reported in Japan. In the present study, the clinicopathological characteristics of 75 samples, comprising 68 cases of Kaposi's sarcoma, 5 cases of primary effusion lymphoma, and 5 cases of multicentric Castleman's disease were investigated. All of these cases were positive for KSHV by immunohistochemistry or PCR analysis. All fifty-two of the AIDS-associated Kaposi's sarcoma cases were males, whereas 7 of the 13 non-AIDS-associated Kaposi's sarcoma cases were females. The mean age of patients with AIDS-associated Kaposi's sarcoma or primary effusion lymphoma was 46 years, whereas the mean age of patients with non-AIDS-associated Kaposi's sarcoma or primary effusion lymphoma was 71.8 and 97.5, respectively. KSHV genotypes were determined based on the sequence of variable region 1 in the K1 gene. Genotypes A and C of KSHV were detected in both AIDS- and non-AIDS-associated Kaposi's sarcoma. Genotype A was detected more frequently in AIDS-associated cases than non-AIDS-associated cases, suggesting that genotype C is broadly distributed in Japan, and genotype A spreads among AIDS patients. Genotype D was detected only in non-AIDS-associated Kaposi's sarcoma. These data confirmed the difference between AIDS- and non-AIDS-associated KSHV diseases with regard to age of onset, gender, and genotypes in Japan. J. Med. Virol. 82:400-406, 2010. (c) 2010 Wiley-Liss, Inc.
收起
摘要 :
OBJECTIVE: Fontan completion in patients with atrial isomerism, in which the inferior vena cava (IVC) and the hepatic vein (HV) drain separately, is technically challenging. Herein, we review our surgical approach to these patient...
展开
OBJECTIVE: Fontan completion in patients with atrial isomerism, in which the inferior vena cava (IVC) and the hepatic vein (HV) drain separately, is technically challenging. Herein, we review our surgical approach to these patients. METHODS: The medical records of 50 consecutive patients with atrial isomerism who underwent Fontan completion between 1998 and 2008 were reviewed retrospectively. RESULTS: Separate HV drainage was present in 17 patients. Patients with interrupted IVC were excluded. Patient characteristics were as follows: median age, 26 months (range 15-149); median weight, 9.6 kg (range 8.1-47.2); right atrial isomerism, 16 patients; and left atrial isomerism, one. The IVC and the separate HV at the level of diaphragm were contralateral in 16 patients, and ipsilateral in one. The surgical procedures for directing blood flow from the IVC and the separate HV to the pulmonary arteries were as follows: en bloc resection of the IVC and the HV and anastomosing these veins to an extracardiac conduit in 10 patients; connecting the IVC to the HV in a side-to-side fashion before anastomosing them to an extracardiac conduit in one; and lateral tunnel in another. When the IVC and the HV were widely separated by the vertebrae, we chose an intra-extracardiac conduit (intra-atrial septation) in four patients and an extracardiac conduit for the IVC and the right HV and lateral tunnel for the separate left HV in one. There was no mortality. Five re-operations were performed (pacemaker in two patients; one each of fenestration, release of outflow obstruction and ligation of collateral arteries). Sixteen patients underwent follow-up catheterisation, which revealed central venous pressure of 12.0 + or - 2.0 mmHg and arterial oxygen saturation of 92% + or - 6%. CONCLUSIONS: The mid-term results of the Fontan completion in patients with atrial isomerism and separate HV drainage were excellent. The distance between the IVC and the separate HV and the position of the vertebrae should be considered when choosing a surgical technique.
收起
摘要 :
OBJECTIVE: This study aimed to determine the expression of the Cap43 gene in supraglottic laryngeal squamous cell carcinoma, and to evaluate any correlation between Cap43 gene expression and tumour-associated macrophage infiltrati...
展开
OBJECTIVE: This study aimed to determine the expression of the Cap43 gene in supraglottic laryngeal squamous cell carcinoma, and to evaluate any correlation between Cap43 gene expression and tumour-associated macrophage infiltration. METHODS: Four human head and neck squamous cell carcinoma cell lines were cultured (Hep2, KB, Ca9-22 and HSC-3) and expression of the Cap43 gene was analysed by Western blotting. In addition, paraffin-embedded samples of supraglottic laryngeal squamous cell carcinoma and normal supraglottic laryngeal mucosa from 84 patients were analysed immunohistochemically using antibodies to Cap43 and cluster of differentiation 68 glycoprotein. Patients' clinical status was compared with their immunohistochemical results. RESULTS: All four head and neck squamous cell carcinoma cell lines exhibited Cap43 expression. The Hep2, Ca9-22 and HSC-3 cells showed a markedly higher level of Cap43 protein than the KB cells. A statistically significant difference was found in Cap43 expression, comparing different differentiation levels and comparing different metastasis stages, for supraglottic squamous cell carcinoma. The number of tumour-associated macrophages correlated with expression of Cap43, not only in the tumour area (r = 0.3708, p = 0.0005) but also in the peritumour area (r = 0.2847, p = 0.0087). CONCLUSION: In supraglottic laryngeal squamous cell carcinoma, overexpression of the Cap43 gene is associated with tumour differentiation and acts an important suppressive factor in the process of tumour metastasis. The Cap43 gene may be a cancer-specific marker. High expression of the Cap43 gene appeared to correlate with infiltration of tumour-associated macrophages.
收起
原文获取