The prevalence of infectious disease in patients with rheumatoid arthritis (RA) is high. Infection causes problems in RA treatment and also affects prognosis. In this study, we examined the clinical characteristics of patients with RA who required hospitalization for infection at our hospital. Between April 2007 and March 2012, there were 79 patients with RA (64 women, 15 men) who had at least one infection requiring hospitalization. Infection most commonly occurred in the upper respiratory tract and pulmonary system (52.3%), urinary tract (11.9%), and skin and connective tissue (11.0%). Compared with the control group (noninfected outpatients with RA), the infection group had a significantly higher incidence of hypoalbuminemia (p < 0.001). Because hypoalbuminemia was already present before infection in the infection group, it can be inferred that hypoalbuminemia is associated with occurrence of infection in patients with RA. Twenty patients had a recurrent infection (comprising a total of 42 infective episodes), while the other 59 had a single infection. Interstitial lung disease (p = 0.029) and daily prednisolone use (p = 0.046) were significantly more common in the recurrent infection group than in the single infection group. These results suggest that hypoalbuminemia, interstitial lung disease, and daily prednisolone use are worth paying attention to with regard to development of infection during RA treatment.
Background: A clinically significant tumor marker is important for the diagnosis of colorectal cancer (CRC) and its management after the treatment. Laminin (Ln)-γ2 is expressed in various types of malignant carcinomas, indicating that it may be a potential serum biomarker. We recently developed a quantitative ELISA assay to determine serum Ln-γ2 levels using the monoclonal antibody to human Ln-γ2 selectively. Methods: Using our previously developed ELISA assay, we examined the serum Ln-γ2 levels in 78 serum specimens from 51 healthy volunteers, 8 patients with benign disease, and 19 CRC patients. Using chemiluminescent immunoassays, we measured carcinoembryonic antigen (CEA) and CA19-9 simultaneously and compared the diagnostic value of a combination of Ln-γ2 with each of these markers. Results: The medians of the serum levels of Ln-γ2 in healthy controls, benign disease patients, and CRC patients were 241.9 pg/mL, 138.8 pg/mL, and 323.0 pg/mL, respectively. Significantly higher Ln-γ2 levels were observed in patients with CRC than in the other two groups. The optimal cutoff value for Ln-γ2 that would distinguish CRC cases from non-malignant cases was 315.8 pg/mL. We observed elevated Ln-γ2 levels (>315.8 pg/mL) in 57.9% of the patients with CRC. The positive rate in patients with CRC for the combination of Ln-γ2 and CEA was 78.9%, higher than that for either of those markers alone (57.9% and 52.6%). The positive rate in CRC patients was high for all markers in advanced stage CRC, but that for CEA and CA19-9 in Stage I/II was low. On the other hand, the positive rate for Ln-γ2 in Stage I/II was high at 50.0%. Conclusions: Serum Ln-γ2 may be a novel biomarker for CRC that can be used along with existing tumor markers for CRC detection.
Germline mutations of BRCA1 cause familial breast and ovarian cancer. The mechanism responsible for the tissue specificity is unknown, although it is considered that estrogen receptor α (ERα) plays a critical role. However, lack of a diploid cell line expressing ERα has prevented researchers from analyzing the impact of ERα on generating genomic instability in the cells with BRCA1 deficiency. To overcome this problem, here we established BRCA1 deficient and ERα positive human breast cell line using biogenetic technology. First we generated doxycyclin (Dox)-inducible BRCA1-defective cells from MCF10A, an ERα-negative normal breast cell line. CS-RfA-ETBsd-shBRCA1 lentiviral vector was generated from oligonucleotides responsible for shRNA sequence in BRCA1 by Gateway recombination from pENTR4-H1tetOx1 entry vector. MCF10A-shBRCA1 cells were established from CS-RfA-ETBsd-shBRCA1-lentivitral infected MCF10A cells with blasticidin selection. Next we generated Dox-inducible ERα-expressing cell line from MCF10A cells by infection of CSIV-TRE-Rfa-Ubc-puro-ERα lentivirus followed by puromycin selection (MCF10A-ERα cells). Finally MCF10A-ERα-shBRCA1 cell line was established by infection of MCF10A-ERα cells with CS-RfA-ETBsd-shBRCA1 followed by double selection with blasticidin and puromycin. The effective inhibition of BRCA1 expression and simultaneous expression of ERα was verified by immunoblotting. Interestingly, whereas ERα expression had no effect on the proliferation of BRCA1 expressing cells, it dramatically suppressed the proliferation of BRCA1-defective cells. Thus, we established a human breast cell line with conditional inhibition of BRCA1 with simultaneous expression of ERα by Dox. This material would be valuable for analyzing ERα-induced genomic instability specifically occurring in a BRCA1-deficient genetic background.
Purpose: To investigate the changes in foveal macular thickness using optical coherence tomography (OCT) in patients with diabetic macular edema and to investigate the role of pars plana vitrectomy. Methods: This study included a total of 24 eyes of 22 patients who underwent pars plana vitrectomy for diabetic macular edema at our hospital from June 2009 to December 2011 and subsequent follow-up examinations for at least 6 months. The patients’ eyes were divided into three groups: those with cystoid macular edema (7 eyes), those with concomitant serous retinal detachment (7 eyes), and those with concomitant epiretinal membrane (10 eyes). OCT was used to investigate the structural changes in diabetic macular edema. Results: Foveal macular thickness in the cystoid macular edema group was 640 ± 189 µm preoperatively and 319 ± 122 µm at 6 months postoperatively. In this group, macular edema had improved (foveal macular thickness <250 µm) in three cases (42%) and residual cystoid macular edema had not completely disappeared from all cases (0%), as evaluated at 6 months postoperatively. Foveal macular thickness in the serous retinal detachment concomitant group was 583 ± 230 µm preoperatively and 348 ± 164 µm at 6 months postoperatively. Macular edema had improved in three cases (42%), cystoid macular edema had disappeared in three cases (42%), and serous retinal detachment had disappeared in all cases (100%) at 6 months postoperatively. Foveal macular thickness in the epiretinal membrane concomitant group was 559 ± 80 µm preoperatively and 263 ± 82 µm at 6 months postoperatively. Macular edema had improved in seven cases (70%) and cystoid macular edema had disappeared in seven cases (70%) at 6 months postoperatively. Conclusions: The results suggest that pars plana vitrectomy is effective for serous retinal detachment, which is generally considered to be difficult to treat with ranibizumab therapy. The majority of the eyes in the epiretinal membrane concomitant group exhibited successful improvement in foveal macular thickness by pars plana vitrectomy.
HERC proteins comprise RCC-like domain that presumably possesses Ran-GEF activity and a HECT domain that contains ubiquitin E3 ligase activity. HERC1 and HERC2 are giant proteins comprising more than 4800 amino acids, and are structurally similar to multiple RCC-like domains and a C-terminal HECT domain. We previously showed that HERC2 is an E3 that targets the familial breast and ovarian cancer susceptibility gene product BRCA1 for degradation, controls G2/M checkpoint of the cell cycle, and regulates DNA replication through interaction with Claspin. However, many functions of HERC2 remain to be clarified. To further investigate the HERC2 functions, we generated a specific antibody to HERC2, and established cell lines in which both HERC1 and HERC2 can be inhibited simultaneously, eliminating a possible compensation of HERC2 function by HERC1. We created a recombinant polypeptide encoding C-terminus of HERC2 containing the HECT domain and generated a rabbit polyclonal antibody to the polypeptide. We then established Doxycyclin (Dox)-inducible HERC2-defective cell lines from HeLa, HCT116 or U2OS cells by infecting lentivirus that is comprised of the CS-RfA-ETBsd-shHERC2 vector expressing blasticidin resistant gene, Tet repressor, and HERC2-specific shRNA under H1tetO promoter/operator regulation, after selection with blasticidin. Dox-inducible HERC1/HERC2 double-defective cell lines were then established by infecting a lentivirus with HERC1-specific shRNA and puromycin resistance to the HERC2-defective cell lines after double selection with blasticidin and puromycin. The successful simultaneous inhibition of HERC1 and HERC2 expression after Dox-treatment was verified by immunoblotting. The dual inhibition of HERC1 and HERC2 severely compromised the retention of Replication protein A (RPA) at ionizing irradiation-induced DNA damage loci whereas single inhibition of HERC1 or HERC2 demonstrated minimal effects. Hence, cell lines with Dox-inducible dual inhibition of HERC1 and HERC2 were established. The cells would contribute to further analyses of the roles of HERC1 and HERC2 in cell-cycles and DNA damage response.
Our previous studies have demonstrated that antenatal glucocorticoid (GC) administration leads to increases of several cardiac function-related proteins and the activation of several enzymes in the process of ATP synthesis in the premature hearts of fetal rats. In addition, the increase in wall thickness of the fetal heart was observed, but the reason remains uncertain. Akt-1 plays a central role in regulating both pathological and physiological cardiac hypertrophy. The aim of the present study is to clearify the expressions of Akt sequential protooncogene involved in the increase of wall thickness in fetal and neonatal hearts with antenatal GC administration. Dexamethasone (DEX) was administered to pregnant rats for two days from day17 or day 19 and 21 of gestation, and the hearts of 19- and 21-day fetuses and 1-day-old neonates were analyzed. Akt in the 19-day fetal heart was significantly lower than that of the neonatal hearts. DEX administration in 19-day and 21-day fetal rats significantly increased compared with the respective controls. Significant increases in c-Myc protein levels were observed in cardiomyocytes of fetuses and neonates by Western blotting and immunohistochemistry. In addition, c-Myc mRNA levels significantly increased in DEX-treated 1-day neonatal primary cardiomyocytes and were inhibited by the precultured glucocorticoid receptor antagonist, RU486. These outcomes were mediated through the GC receptor in the heart. We found that antenatal GC administration significantly increases the expressions of Akt-1 and c-Myc in fetal rat hearts. These results suggest that these proteins may contribute to cardiac enlargement of the premature heart and lead to cardiac growth.
Coronary computed tomography (CT) angiography uses advanced CT technology to detect not only coronary artery stenosis but also regional wall-motion abnormalities. We present a case of severe diffuse stenosis in the left anterior descending artery successfully treated with coronary CT angiograms. We report the ways that cardiac CT angiography was useful for diagnosing coronary artery stenosis in the present case, as follows: 1) Coronary CT angiography was useful in the use of wires during percutaneous coronary intervention, even for severe and diffuse stenotic lesions; 2) coronary CT angiography depicted the abnormality in the left ventricular anterior wall, which was consistent with the stenotic lesion in the left anterior descending artery; 3) wall thickness was within the normal range, although endocardial visualization was poor; and 4) the number of visible CT slices in the left anterior descending artery was decreased in the stenotic lesion and increased in the peripheral lesion. The present case demonstrates that coronary CT angiography before catheterization is useful in percutaneous coronary intervention and enabling an accurate diagnosis, particularly for cardiac wall motion and cardiac enhancement. Post-processing and offline analysis form a significant part of the time taken to report studies, and an efficient method of providing quantitative reports is nencessary. Thus, collaboration among radiologists, technicians, and cardiologists is essential to provide successful treatments.
[Introduction] When a patient with hemophilia develops thrombosis, it makes the choice of treatment difficult because of their contradictory condition. We experienced the case of a patient with hemophilia A who underwent successful percutaneous coronary intervention (PCI) with replacement therapy of clotting factor VIII (FVIII) concentrate and anticoagulation therapy with unfractionated heparin (UFH). [Case] A 64-year-old man with mild hemophilia A and an attack of chest pain was transferred to our hospital. We diagnosed him with acute myocardial infarction (AMI) and then performed coronary angiography after 3,000 IU FVIII concentrate was administered to achieve a peak level of 1.0 IU mL−1; 100% and 90% stenosis was revealed in lesion #7 and #2, respectively, during this procedure. The #7 lesion was treated immediately with PCI performed under the administration of UFH. After PCI, antiplatelet therapy was started. During UFH treatment, we administered FVIII concentrate to achieve a trough level of 0.5–0.6 IU mL−1. On day 16, PCI for lesion #2 with 90% stenosis was performed under the administration of FVIII concentrate and UFH in the same way. Then antiplatelet therapy was continued, and regular replacement of FVIII concentrate was started to prevent bleeding. The patient did not experience any hemorrhagic complication or restenosis of the treatment sites. [Discussion] The number of patients with hemophilia complicated with AMI may increase accompanied by the increase in the aged patients. An appropriate treatment for AMI in patients with hemophilia should be considered by an accumulation of the therapeutic experience of these patients.