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To inhibit arthritis upstream of inflammatory cytokine release and matrix metalloproteinase (MMP) action, we designed de novo a small-molecule inhibitor of c-Fos/activator protein-1 (AP-1) using three-dimensional (3D) pharmacophore modeling. This model was based on the 3D structure of the basic region-leucine zipper domain of AP-1-DNA complex. Administration of this inhibitor prevented type II collagen-induced arthritis from day 21, before the onset of arthritis, or from day 27, resolved arthritis after its onset. Suppression of disease was accomplished by reducing the amounts of inflammatory cytokines and MMPs in vivo in sera and joints and in vitro in synovial cell and chondrocyte cultures. The primary action of this molecule was the inhibition of matrix-degrading MMPs and inflammatory cytokines including interleukin 1beta; this molecule also synergized with anti-tumor necrosis factor alpha to inhibit arthritis. Thus, selective inhibition of c-Fos/AP-1 resolves arthritis in a preclinical model of the disease.

患者は67歳の女性, 黄疸と右季肋部痛を主訴に入院した. 精査の結果, Dubin-Johnson症候群 (DJS) に合併した肝細胞癌 (HCC) と診断した. 術前のT. Bil値は3.8mg/dlであったが, 他の肝予備能が保たれていたため肝S5亜区域切除術を施行した. 術後, T. Bil値は5.2mg/dlまで上昇したが漸減し, 第17病日に軽快退院した. 術後13カ月で多発性の肺転移をきたしたがfluorouracil, doxorubicin, cisplatinによるFAP療法を計14コース施行し, 術後5年経過した現在, PRの状態にある. 本邦でDJSにHCCが合併し肝切除が施行された症例は, 今までに8例報告されている. いずれも術前に高ビリルビン血症を伴っていたが肝不全に至らず耐術していた. しかし, 体質性黄疸患者に肝切除を施行する場合, T. Bil値は肝障害度を直接表わす指標でないものの, 高度黄疸下の大量肝切除は黄疸が遷延することがあり注意が必要である.

症例は71歳, 女性. C型肝硬変合併肝腫瘍 (S8), 脾腫による汎血球減少症の診断でS8部分切除・脾摘術を施行した. 第8病日に39℃台の発熱, 下痢がみられたが対症療法で軽快した. しかし第30病日に再び39℃台の発熱, 腹痛が出現し, 腹部CT検査で脾静脈から門脈左右枝におよぶ広範な血栓形成を認めた. ウロキナーゼ (24万単位/日), ヘパリン (5,000単位/日) 静脈内投与による血栓溶解療法を開始し, 第37病日の腹部CT検査で血栓の縮小がみられたため, 以後ワーファリン (2mg/日) 内服による抗凝固療法に変更した. 第69病日の腹部CT検査では血栓は完全に消失し, 第72病日に退院した. 術後113日目の腹部CT検査でも再発はなく, ワーファリンを減量したのち中止した. 脾摘後の門脈血栓症は比較的頻度の高い合併症であるが, 脾摘と肝切除の同時施行での発症例は自験例を含め3例であった. 術後に原因不明の発熱や腹痛などが続いた場合は本病態も考慮し, 腹部超音波検査や腹部CT検査を適宜施行することが重要である.

Background Sepsis is a potentially fatal syndrome mediated by an early [e.g., tumor necrosis factor-alpha (TNF-α)] and late [high mobility group box-1 (HMGB-1)] proinflammatory cytokine response to infection. Sepsis-induced acute kidney injury (AKI) is associated with a high mortality. C-Fos/activator protein-1 (AP-1) controls the transactivation of proinflammatory cytokines via AP-1 binding in the promoter region. T-5224 is a de novo small molecule inhibitor of c-Fos/AP-1 that controls gene expression of multiple proinflammatory cytokines. We investigated whether T-5224, a selective inhibitor of c-Fos/AP-1, improves survival in lethal lipopolysaccharide (LPS)-induced AKI by inhibiting early (TNF-α) and late (HMGB-1) proinflammatory cytokine response. Methods Mice were divided into four groups (control, LPS, LPS + T-5224, and T-5224 only). Control mice were administered polyvinylpyrrolidone (PVP) solution orally, immediately after intraperitoneal (i.p.) saline injection. LPS mice were administered PVP solution orally immediately after i.p. LPS (10 mg/kg) injection. LPS + T-5224 mice were administered T-5224 orally (300 mg/kg) immediately after i.p. LPS injection. T-5224 mice were administered T-5224 orally (300 mg/kg) after i.p. saline injection. Serum concentrations of TNF-α, HMBG-1, and interleukin (IL)-10 were measured by enzyme-linked immunosorbent assay (ELISA). Serum blood urea nitrogen (BUN) and creatinine concentrations were commercially analyzed. Finally, histological examination was performed on the kidney. Results Treatment with T-5224 decreased serum TNF-α and HMGB-1 levels and increased survival after LPS injection. Furthermore, T-5224 treatment decreased serum BUN and creatinine concentrations but increased serum IL-10 concentration. LPS-induced pathological changes in kidney were attenuated by T-5224 treatment. Conclusions These results suggest that T-5224, a selective inhibitor of c-Fos/AP-1, inhibits expression of early and late proinflammatory cytokines, protecting mice from LPS-induced lethality. T-5224 is a potential approach for decreasing lethality in sepsis-induced AKI.

We retrospectively analyzed 7 patients with refractory colorectal cancer treated with TAS-102 as salvage therapy. Subjects were 3 men and 4 women. The median age at initiation of TAS-102 was 71 years (range, 41-82 years). The number of target organs was 1 in 5 patients, 2 in 1 patient, and 3 in 1 patient. The median treatment courses were 2 courses (range, 1-6 courses). The reason for discontinuation was hematological toxicity in 1 patient, patients' wish in 3 patients, disease progression in 2 patients, and worsening of general condition in 1 patient. The median survival time since the first administration of TAS-102 was 9 months.

We analyzed immune nutritional factors that affected the incidence of SSI in rectal cancer surgery.A total of 103 patients who underwent rectal cancer resection were enrolled in this retrospective study. The risk factors (DM, BMI18.5, ≥25.0, PNI≤40, G/L2, CONUT≥2, mGPS D) for SSI (Grade≥Ⅱ) were analyzed.The factors that significantly affected SSI (in 13 cases) was PNI≤40 on univariate analysis. In the analysis adjusted by age and sex, mGPS D and PNI≤40 were significant factors. In the stepwise selection method, PNI≤40 was selected as an independent factor.As a risk factor for SSI after rectal cancer surgery, PNI≤40 and mGPS were risk factors.

Colorectal cancer associated perforation initially develops as pan-peritonitis but easily progresses to septic shock, which can be fatal. As such, it can be hard for patients to recover from this pathological condition. A 79-year-old man who was suffering from pan-peritonitis due to sigmoid colon cancer-associated perforation and also had a metastatic hepatic lesion was admitted to our hospital. He underwent an emergency operation in October 2012. Due to hemodynamic instability, peritoneal lavage and drainage, and stomal formation were performed during the operation. Polymyxin-B direct hemoperfusion (PMXDHP) and continuous hemodiafiltration (CHDF) were performed for septic shock and acute renal failure, respectively. The patient was administered 5 courses of chemotherapy consisting of capecitabine, oxaliplatin, and bevacizumab (Cape+L-OHP +Beva) with no severe adverse reactions; the primary colonic and metastatic hepatic lesions showed a good response to the chemotherapy. A radical resection for the sigmoid colon cancer, including a partial hepatic resection for the metastatic lesion, was performed in May 2013. Surveillance examinations have indicated that the patient is recurrence-free 13 months after radical resection.

切除不能の膵・胆道癌患者に対する病状説明のinformed consent(IC)に際し,家族の関与を積極的に認めるICの有用性を評価し,今後の問題点も検討した.過去7年間に筆者(S.S)が病状説明を行った切除不能の膵・胆道癌89例を対象に,まず家族に対し患者本人に病状を正しく告知することの重要性と支援体制の確立を求めた.この結果,最終的に73人(82%)の患者に病状説明, 41人(46%)に予後の説明を行った.説明後のうつ状態からの回復は比較的早かったが,家族の支援が消極的であったり,キーパーソンが不在である場合は回復が遅れる傾向にあった.我が国では,未だ患者の自己決定権の意識が希薄で,患者自身が意志決定する際にも家族の支援が必要である.今後は,患者の意志を尊重するという考えを個人情報保護法とともに社会一般に啓蒙し,周知させていく努力が必要と考えられる.

To confirm the incidence and risk factors of incisional hernia after colorectal cancer surgery, we analyzed the clinical data including the surveillance computed tomography (CT) examination.One hundred sixty seven patients with open abdominal surgery for colorectal cancer were analyzed retrospectively.Incisional hernia was recognized in 27 cases (16.2%), and occurred at median 7 (1-21) months after surgery. Multivariate analysis showed the risk factors for incisional hernia were female (p=0.0014), distal colon and rectal cancer (p=0.0038), high body mass index (p=0.0055) and lower serum albumin (p=0.0081).Obesity, lower median incision and malnutrition might seem to relate to the incisional hernia after colorectal cancer surgery.

Methotrexate (MTX) is the first-choice drug for rheumatoid arthritis (RA); however, the pharmacodynamics of MTX in Japanese patients with RA treated legitimately according to the government recommended dosage, 6 mg per week, are unknown. Methotrexate and its metabolite, 7-hydroxy MTX (7-OH MTX), were measured in sera of 16 outpatients with active RA in the first week of MTX treatment and 4-12 weeks after the introduction at 0, 1, 2, 4, and 8 h after administration of the first and the third 2-mg capsule, followed by sampling at 48, 96, and 168 h. The mean maximal serum drug concentration (mean C(max)) of MTX attained at 1-2 h after ingestion of the first capsule was 0.215 and 0.252 microM, respectively, in the first and the follow-up week. The mean C(max) after ingestion of the third capsule was 0.223 microM and 0.357 microM. The mean C(max) of 7-OH MTX was 0.0334 and 0.0289 microM for the first capsule, and 0.0495 and 0.0672 microM for the third capsule. The results indicate that MTX does not accumulate or deposit in the body of Japanese patients with RA when treated with 6 mg per week, and pharmacodynamics of MTX are comparable to those in overseas patients treated with 7.5 mg per week.

A 55-year-old woman with well-controlled systemic lupus erythematosus (SLE) suffered from the abrupt onset of massive intractable ascites, which did not respond to conventional diuretic therapy. While treatment with methylprednisolone pulse therapy ameliorated this lupus peritonitis, neuropsychiatric symptoms then appeared. After a diagnosis of the central nervous system (CNS) lupus, pulse therapy was continued and the patient recovered from the lupus psychosis. We discuss the differential diagnosis between CNS lupus and steroid-induced psychosis with particular references to recent diagnostic methods for CNS lupus.

We have studied the effect of low-dose prednisolone administered before sleep on the hypothalamic-pituitary-adrenal axis and the symptoms of patients with rheumatoid arthritis (RA). Plasma adrenocorticotropic hormone (ACTH) and serum cortisol levels were measured in the basal state and after hypoglycemic stress induced by the insulin tolerance test in 21 patients receiving prednisolone at 3-5 mg daily. The patient's global assessment of their disease activity scores on a 100-mm visual analogue scale (VAS) and self-reporting of their functional status using the health assessment questionnaire (HAQ) were evaluated. While both the cortisol and the ACTH responses were impaired dose-dependently in patients treated with prednisolone, the ACTH response was maintained in patients treated with a single daily 3-mg dose of prednisolone before sleep. There was an inverse correlation between the extent of the ACTH response and disease activity as revealed by the VAS (r = 0.521, P0.05). There was also a weak correlation between VAS and the self-rating depression scale (SDS) (r = 0.443), especially when only patients with an HAQ score10 were included in order to exclude any possible contribution of the limitations in the activities of daily living to the SDS score (r = 0.859, P0.05). These results suggest that a single daily low dose (3 mg) of prednisolone administered before sleep maintains the ACTH response in RA patients, and patients with a good ACTH response appear to be less depressed and have milder symptoms.

The transcription factor activator protein-1 (AP-1) is an attractive target for the treatment of immunoinflammatory diseases, such as rheumatoid arthritis. Using the three-dimensional (3D) X-ray crystallographic structure of the DNA-bound basic region leucine zipper (bZIP) domains of AP-1, new cyclic disulfide decapeptides were designed and synthesized that demonstrated AP-1 inhibitory activities. The most potent inhibition was exhibited by Ac-c[Cys-Gly-Gln-Leu-Asp-Leu-Ala-Asp-Gly-Cys]-NH2 (peptide 2) (IC50 = 8 microM), which was largely due to the side chains of residues 3-6 and 8 of the peptide, as shown by an alanine scan. To provide structural information about the biologically active conformation of peptide 2, the structures of peptide 2 derived from molecular dynamics simulation of the bZIP-peptide 2 complex with explicit water molecules were superimposed on the solution structures derived from NMR measurements of peptide 2 in water. These showed a strong structural similarity in the backbones of residues 3-7 and enabled the construction of a 3D pharmacophore model of AP-1 binding compounds, based on the chemical and structural features of the amino acid side chains of residues 3-7 in peptide 2.

We evaluated the clinical usefulness and safety of transradial approach for transcatheter arterial chemoembolization (TACE) in patients with hepatocellular carcinoma (HCC) compared with that of conventional transfemoral approach. The two groups (radial group, n = 177; femoral group, n = 150) of cases were retrospectively compared with regard to the successful rate of angiography or TACE, time required for catheterizaiton and complications. Hepatic angiography and TACE were completed in 174 (98.3%) of 177 cases in the radial group. There was no intergroup difference of time required for catheterization. Minor complications (dull pain, numbness) occurred in 8 (4.6%) patients in the radial group, and there were lower complications in the radial group compared to the femoral group. TACE by our new transradial approach was found to have therapeutic efficacy with lower complications comparable to that of the conventional transfemoral approach.

MCF-2 (DBL) proto-oncogene is a prototype guanine nucleotide exchange factor (GEF) that modulates Rho GTPases such as Rho, Rac, and Cdc42. Although the partial sequence of mouse MCF-2 has been determined, its full-length cDNA and biochemical functions had not been elucidated. We isolated the complete mouse MCF-2 cDNA and obtained recombinant functional protein. Homology between the mouse and human MCF-2 (DBL) cDNAs is 75.08% identity and between the mouse and human amino acid sequences 74.52% identity. Analysis of tissue distribution showed that mouse MCF-2 mRNA is expressed in brain, kidney, intestine, and testis. The brain-specific transcript is an alternatively spliced derivative that omits the 48 bp exon 11. A similar alternatively spliced mRNA product is also found in humans (DBL). Guanine nucleotide exchange activities of the testis-expressed mouse Mcf-2 and human Dbl were analyzed using RhoA, Rac1, and Cdc42 as substrates. RhoA and Cdc42 were activated similarly by both gene products, but Rac1 was activated only by the mouse product. The brain-specific Mcf-2 gene product, and its human counterpart, was less active than the respective testis-specific products. This indicates that the element encoded by the 48 bp exon missing in the brain transcripts is necessary for full GEF activity. This report provides fundamental data on the structure of Mcf-2, which regulates a variety of cellular signaling pathways.

The compound 26,26,26,27,27,27-F(6)-1alpha,25(OH)(2)D(3) is a hexafluorinated analog of the active form of Vitamin D(3). The enhanced biological activity of F(6)-1alpha,25(OH)(2)D(3) is considered to be related to a decreased metabolic inactivation of the compound in target tissues such as the kidneys, small intestine, and bones. Our previous study demonstrated that CYP24 is responsible for the metabolism of F(6)-1alpha,25(OH)(2)D(3) in the target tissues. In this study, we compared the human and rat CYP24-dependent metabolism of F(6)-1alpha,25(OH)(2)D(3) by using the Escherichia coli expression system. In the recombinant E. coli cells expressing human CYP24, bovine adrenodoxin and NADPH-adrenodoxin reductase, F(6)-1alpha,25(OH)(2)D(3) was successively converted to F(6)-1alpha,23S,25(OH)(3)D(3), F(6)-23-oxo-1alpha,25(OH)(2)D(3), and the putative ether compound with the same molecular mass as F(6)-1alpha,25(OH)(2)D(3). The putative ether was not observed in the recombinant E. coli cells expressing rat CYP24. These results indicate species-based difference between human and rat CYP24 in the metabolism of F(6)-1alpha,25(OH)(2)D(3). In addition, the metabolite with a cleavage at the C(24)z.sbnd;C(25) bond of F(6)-1alpha,25(OH)(2)D(3) was detected as a minor metabolite in both human and rat CYP24. Although F(6)-1alpha,23S,25(OH)(3)D(3) and F(6)-23-oxo-1alpha,25(OH)(2)D(3) had a high affinity for Vitamin D receptor, the side-chain cleaved metabolite and the putative ether showed extremely low affinity for Vitamin D receptor. These findings indicate that human CYP24 has a dual pathway for metabolic inactivation of F(6)-1alpha,25(OH)(2)D(3) while rat CYP24 has only one pathway. Judging from the fact that metabolism of F(6)-1alpha,25(OH)(2)D(3) in rat CYP24-harboring E. coli cells is quite similar to that in the target tissues of rat, the metabolism seen in human CYP24-harboring E. coli cells appear to exhibit the same metabolism as in human target tissues. Thus, this recombinant system harboring human CYP24 appears quite useful for predicting the metabolism and efficacy of Vitamin D analogs in human target tissues before clinical trials.

Features characteristic to rheumatoid arthritis (RA) including synovial overgrowth and joint destruction are experimentally produced by augmenting c-fos gene expression. We show that cyclin dependent kinase inhibitor p21waf1/cip1, that inhibits cell proliferation, is down-regulated in conjunction with up-regulation of c-fos in the lymphocytes of patients with RA. As to the mechanism of down-regulation of p21waf1/cip1 gene expression, transfection studies in U937 cells showed that c-fos down-regulated phosphorylation and dimerization of signal transducers and activators of transcription (STAT) 1, thereby inhibiting interferon γ-induced transactivation of p21waf1/cip1. Phosphorylation of STAT1 was indeed decreased in the lymphocytes of patients with RA. Thus, under overexpression of c-fos gene, c-Fos inactivates STAT1 to down-regulate p21waf1/cip1 gene expression in the lymphocytes of patients with RA, and in this way may enhance proliferation of lymphocytes.

In terms of the relationship between synovial inflammation and radiographic changes, including both joint damage repair and progression,1 in rheumatoid arthritis (RA), pre-existing joint damage and persistent synovitis may promote joint destruction, while in the absence of synovitis, damaged joints may heal.2 ,3 Although presentation of radiographic results using cumulative probability plots has substantially improved understanding of clinical trial data,4 the effects of treatments on radiographic progression and improvement (regression) in individual RA patients has not yet been fully explained. In the JESMR study,5 ,6 151 active RA patients unresponsive to treatment with methotrexate (MTX) were randomised into 1 of 2 treatment groups: etanercept (ETN) 50 mg/week with 6–8 mg/week of MTX (the E+M group), or ETN alone (the E group). Radiographs of the hands and feet before ETN (baseline) and during the first year of treatment were available from 53 (72%) and 68 (88%) patients in the E and E+M groups, respectively. Baseline characteristics of patients were comparable between those with and without available radiographic data in each treatment group (data not shown). However, most patients without data did not complete the study up to Week 52 as per protocol, chiefly due to lack of efficacy in the E group.6 The mean baseline total Sharp-van der Heijde score (TSS)7 was 114.5 in the E group and 113.1 in the E+M …

The DBL (MCF-2) proto-oncogene is a prototype guanine nucleotide exchange factor (GEF) that modulates the Rho family of GTPases. In this communication we describe the isolation of three novel splicing variants of Dbl. The prototype Dbl gene (designated var.1 here) contains 25 exons, while splicing variant 2 (var.2) lacks exons 23 and 24. Var.3 contains additional 3 exons from 5(')-UTR in place of exon 1, while var.4, var.2, and var.3 contain a 48bp insertion between exons 10 and 11, resulting in the insertion of 16 amino acids. We found that var.1 was expressed only in brain, whereas var.3 was expressed in heart, kidney, spleen, liver, and testis, and var.4 in brain, heart, kidney, testis, placenta, stomach, and peripheral blood. The Dbl protein was detectable in brain, heart, kidney, intestine, muscle, lung, and testis. An assay for GEF activity revealed that the var.2 exhibits decreased GEF activity towards Cdc42, var.3 exhibits a weak but significant activity toward Rac1 and Cdc42, var.4 exhibits significant activity toward RhoA and Cdc42, while var.1 exhibits no activity toward RhoA, Rac1, or Cdc42. In summary, we describe 4 splicing variants of the human DBL proto-oncogene that show different tissue distributions and GEF specificities.

The incidence of postoperative complications and mortality are usually higher in patients with preoperative malnutrition. Malnutrition often preexists, particularly in patients undergoing surgery for esophageal cancer, which is substantially invasive. It is therefore important to understand the nutritional condition of patients and actively control perioperative nutrition.Our hospital has been providing nutritional status screening for patients before resection of esophageal cancer, and we report the current status and evaluation results in this article.This screening included 158 patients requiring radical resection of esophageal cancer.Age, comorbidity with diabetes, body mass index(BMI), serum albumin(Alb), Onodera's prognostic nutritional index(PNI), and Glasgow prognostic score(GPS)were used as nutritional indicators to stratify patients for analysis.Evaluation parameters included the incidence of postoperative complications(any complication, pulmonary complications, psychiatric disorder, and anastomotic leakage)and rates of long-term postoperative hospitalization.The analysis indicated that age, BMI, serum Alb, PNI, and GPS are useful for predicting the onset of postoperative complications and prolonged postoperative hospitalization.For such patients, more active nutritional control should be provided.

Stoma creation is one of the risk factors for the incisional surgical site infection (SSI) which can develop the patient's pain in a colorectal surgery.We performed the subcuticular suture with subcutaneous negative pressure drainage and sealing with liquid tissue adhesive for the prevention of wound infection at the stoma creation.A total of 72 patients between January 2006 and December 2012 were retrospectively analyzed. Up to December 2008, the wound closure was performed by the percutaneous transdermal interrupted suture with monofilament nylon sutures (conventional procedure). From January 2009, the 10-Fr silastic flexible drains were placed at the subcutaneous space and subcuticular suture using a monofilament absorption string was performed. A liquid tissue adhesive was used to seal the skin wound (revised procedure). There was no difference between the conventional group and the revised group in age and gender. Risk factors in two groups were not found the significant difference except diabetes mellitus. Incisional SSI was observed in 23 patients out of 72 patients (31.9%). There was no significant difference in incidence in clinicopathological factors. Only the revised procedure of wound closure significantly decreased 13.8% of incisional SSI rate from 44.2% in the conventional procedure.Our several changes of wound closure including tissue adhesive, subcuticular suture and subcuticular closed suction drainage reduced incisional SSI.

There are very few reports of esophageal G-CSF-producing cancer. This report describes a case of G-CSF-producing esophageal squamous cell carcinoma we recently encountered. A 70-year-old male patient had Stage III esophageal squamous cell carcinoma. The patient received preoperative chemotherapy, and therapeutic response for the primary lesion was rated as complete response and that of the lymph node metastasis as stable disease. A radical operation was then performed. A relapse to neutrophilia occurred as liver metastasis recurred postoperation, and serum G-CSF level was high. Immunohistochemical staining of the resected specimen with anti-G-CSF antibody was positive. The patient died about 1 year after the operation. According to our search of the literature, there are 22 cases of esophageal G-CSF-producing cancer. Carcinosarcoma was more frequent as compared to esophageal non-G-CSF-producing cancer. The prognosis was graver in those cases of G-CSF-producing squamous cell carcinoma, relative to cases of non-G-CSF-producing esophageal squamous cell carcinoma.

Our recent studies into the role of autoantibody-inducing CD4 T cells in autoimmune disease have necessitated studies on the mechanism of TCR revision, a phenomenon that has been difficult to approach experimentally. Here we describe a detailed experimental technique to investigate the molecular events involved in TCR revision.

There are many issues with animal models that represent human autoimmune disease or protocols to induce systemic autoimmunity, especially protocols to induce disease in normal mice not having a genetic disposition to autoimmunity. We describe here a novel and completely reproducible experimental technique that can induce systemic autoimmunity or systemic lupus erythematosus (SLE) in mice otherwise not prone to spontaneous autoimmune disease. This protocol involves the repeated immunization of mice with the same antigen. This rather simple technique enables us to perform exact and quantitative in vivo animal experiments with great accuracy.

Among the methods used in molecular biology, in vitro biochemical assays are more common, whereas in vivo assays, including the use of animal models, are less widely employed. In our studies on systemic lupus erythematosus (SLE), we have identified a novel T cell subtype termed "autoantibody-inducing CD4 T cells" (aiCD4 T cell) that is responsible for the development of autoimmunity. In order to identify and isolate these cells, we developed a new technique that involves the transfer of candidate T cell subpopulations into naïve mice and assaying for the development of autoantibodies in the recipient mice. We have previously described an experimental system in which mice not normally prone to autoimmune diseases can be induced to develop experimental SLE. In this experimental system, autoantibody-inducing CD4 T (aiCD4 T) cells are generated via de novo T cell receptor revision at peripheral lymphoid organs. These aiCD4 T cells not only induce a variety of autoantibodies but also promote the final differentiation of CD8 T cells into cytotoxic T lymphocytes, resulting in a pathology identical to SLE. We needed to develop a new methodology to isolate this subpopulation of aiCD4 T cells. Here we describe an in vivo assay to detect and isolate aiCD4 T cells by transferring the candidate cells into naïve recipient mice and monitoring the production of the appropriate antibody or cytokine.

Case1: A 63-year-old woman with diarrhea and hematochezia was diagnosed as having rectal cancer invading the pelvis. Six courses of the 5-fluorouracil, leucovorin, and oxaliplatin( mFOLFOX6) plus panitumumab regimen were administered after sigmoid colostomy, following which low anterior resection was performed. Since the 6 courses of mFOLFOX6 were administered postoperatively, no evidence of recurrence has been observed for 18 months. Case2: A 52-year-old man with high fever and abdominal pain was diagnosed as having rectal cancer invading the bladder with a vesicorectal fistula. After transverse colostomy and 6 courses of mFOLFOX6 plus panitumumab, high anterior resection with partial cystectomy was performed. Since the 8 courses of capecitabine plus oxaliplatin (XELOX) were administered postoperatively, no evidence of recurrence has been observed for 12 months. Although no consensus has been reached pertaining to the use of neoadjuvant chemotherapy for the treatment of colorectal cancer, we could, in this study, demonstrate the efficacy of neoadjuvant chemotherapy with panitumumab for the treatment of locally advanced colorectal cancer.

We examined the association of the physical, nutritional, and immune status with adverse events in patients treated with mFOLFOX6 adjuvant chemotherapy after colorectal cancer surgery.This study included 17 patients, 7 male and 10 female. The median age was 62( range, 32-75) years. The median number of treatment cycles was 12 (range, 4-12). Age, performance status( PS), body mass index( BMI),serum albumin level( Alb),Onodera's prognostic nutritional index( PNI), controlling nutritional status( CONUT),Glasgow prognostic score( GPS),the granulocyte/lymphocyte ratio( G/L),neutrophil count, and the total lymphocyte count( TLC) were evaluated with regard to the nutrition and immunity status of the host before chemotherapy. The incidents of toxicity of greater than Grade 2 severity, excluding gastrointestinal events or gastrointestinal toxicities, were analyzed to determine the correlation with host status.Any toxicities and toxicities without digestive symptoms were observed in 11 patients( 64.7%),and the number of incidents was significantly increased in patients with a PNI of45. Gastrointestinal toxicities were observed in 4 patients (23.5%), but there were no significant correlations with any of the factors investigated.Toxicities are observed to a greater extent in patients with a PNI of45 during adjuvant chemotherapy. These findings suggest that nutritional support may be required to safely administer mFOLFOX6 adjuvant chemotherapy.

We report herein the usefulness of interventional radiological treatment( IVR) for hepatocellular carcinomas( HCCs), based on the results of transcatheter arterial chemoembolization( TACE) and transarterial infusion( TAI).The study included 256 cases of HCC. TACE and TAI were performed for durations permitted by the degree of liver damage. Results(: 1) TACE was performed in 224 cases( average: 4.5 times, range: 1-14 times), and TAI was performed in 32 cases( average: 2.3 times, range: 1-8 times).( 2) The 3- and 5-year survival rates for all cases were 45.5% and 31.6%, respectively.( 3) We classified all cases according to the number of HCCs, solitary, 2-4, and multiple HCCs, and found no significant differences in the survival rate between the 3 groups( p=0.207),( 4) TAI was followed by TACE in non-responsive cases, and the median survival time of the TAI group was 8.5 months.We can expect benefits from repeated TACE treatment in the multiple HCCs group, compared to the solitary HCC group. TAI followed by TACE might improve the prognosis of unresectable and recurrent HCCs. Therefore, we conclude that IVR has clinical benefit as local treatment for HCC.

Although hepatic resection is the most effective therapy for patients with liver metastasis from colorectal carcinoma, a subset of patients cannot undergo surgical treatment for several reasons, including age-related general health decline or poor conditions associated with coexisting diseases, even if the lesions are resectable. A 75-year-old man with a recurrent lesion in the liver underwent right hemicolectomy and partial hepatic resection to treat colonic cancer and a liver metastasis, followed by uracil and tegafur plus Leucovorin( UFT+LV) as adjuvant chemotherapy at 6 months after the initial surgery. Although the lesion was resectable, the patient preferred chemotherapy to surgery, and capecitabine plus oxaliplatin plus bevacizumab was administered; however, the treatment was stopped in the middle of the second course because of oxaliplatin -related toxicities. Capecitabine plus bevacizumab was introduced as the following chemotherapy regimen, and no adverse reactions were observed during this therapy. After 5 courses of administration, the lesion disappeared on CT examination, and no new lesions were found after 9 courses. Thus, the treatment response was classified as complete response (CR) and remains as such after 13 courses.

Patients often experience decreased oral intake due to primary systemic therapy (DCF [docetaxel, cisplatin, and fluorouracil ] therapy) administered during the treatment of esophageal carcinoma; measures to cope with this problem have been sought. We therefore examined the relationship between the presence or absence of decreased oral intake and blood biochemistry( serum albumin[ Alb] level, white blood cell[ WBC] count, neutrophil count, and serum sodium[ Na] level) during the 12 courses of DCF therapy administered as primary systemic therapy to 6 patients with esophageal carcinoma. Decreased oral intake occurred frequently from day 6 to day 12 after the initiation of DCF therapy. During this period, decreased serum Alb levels were observed in patients with decreased oral intake but not in patients without decreased oral intake. The incidence of decreased oral intake was 100% in patients whose serum Alb levels decreased to3.5 g/dL, but it did not exceed 33.3% in patients whose serum Alb levels were ≥3.5 g/dL. The serum Na level, WBC count, and neutrophil count were less affected than the serum Alb level, suggesting that decreased oral intake was associated with decreased serum Alb level.

This study was conducted to analyze the outcomes of endoscopic stent placement (n=9) and bypass surgery (n=9) with regard to perioperative complications and dietary intake conditions in patients with unresectable advanced gastric cancer with stenosis. Regarding perioperative complications, 1 patient in the stent group experienced a stent failure and 1 patient in the bypass group developed an adhesive ileus. Dietary intake began from the first day in the stent group and from the fourth day in the bypass group, and it was continued for 55 and 113 days, respectively. There was no difference in the introduction of chemotherapy or length of treatment between the groups, and the survival period for the patients in the stent and bypass groups was 83 and 127 days, respectively. Endoscopic stent placement for unresectable advanced gastric cancer with stenosis is a safe and effective method for improving the quality of life( QOL) of patients.

Our recent studies into the role of autoantibody-inducing CD4 T cells in autoimmune disease have necessitated studies on the mechanism of TCR revision, a phenomenon that has been difficult to approach experimentally. Here we describe a detailed experimental technique to investigate the molecular events involved in TCR revision. + T cell (aiCD4 + T cell), T cell receptor (TCR) revision, Recombination-activating gene (RAG), TCR basal signaling

There are many issues with animal models that represent human autoimmune disease or protocols to induce systemic autoimmunity, especially protocols to induce disease in normal mice not having a genetic disposition to autoimmunity. We describe here a novel and completely reproducible experimental technique that can induce systemic autoimmunity or systemic lupus erythematosus (SLE) in mice otherwise not prone to spontaneous autoimmune disease. This protocol involves the repeated immunization of mice with the same antigen. This rather simple technique enables us to perform exact and quantitative in vivo animal experiments with great accuracy.

Among the methods used in molecular biology, in vitro biochemical assays are more common, whereas in vivo assays, including the use of animal models, are less widely employed. In our studies on systemic lupus erythematosus (SLE), we have identified a novel T cell subtype termed "autoantibody-inducing CD4 T cells" (aiCD4 T cell) that is responsible for the development of autoimmunity. In order to identify and isolate these cells, we developed a new technique that involves the transfer of candidate T cell subpopulations into naive mice and assaying for the development of autoantibodies in the recipient mice. We have previously described an experimental system in which mice not normally prone to autoimmune diseases can be induced to develop experimental SLE. In this experimental system, autoantibody-inducing CD4 T (aiCD4 T) cells are generated via de novo T cell receptor revision at peripheral lymphoid organs. These aiCD4 T cells not only induce a variety of autoantibodies but also promote the final differentiation of CD8 T cells into cytotoxic T lymphocytes, resulting in a pathology identical to SLE. We needed to develop a new methodology to isolate this subpopulation of aiCD4 T cells. Here we describe an in vivo assay to detect and isolate aiCD4 T cells by transferring the candidate cells into naive recipient mice and monitoring the production of the appropriate antibody or cytokine.

M-phase promoting factor is a complex of cdc2 and cyclin B that is regulated positively by cdc25 phosphatase and negatively by wee1 kinase. We isolated the wee1 gene promoter and found that it contains one AP-1 binding motif and is directly activated by the immediate early gene product c-Fos at cellular G(1)/S phase. In antigen-specific Th1 cells stimulated by antigen, transactivation of the c-fos and wee1 kinase genes occurred sequentially at G(1)/S, and the substrate of wee1 kinase, cdc2-Tyr15, was subsequently phosphorylated at late G(1)/S. Under prolonged expression of the c-fos gene, however, the amount of wee1 kinase was increased and its target cdc2 molecule was constitutively phosphorylated on its tyrosine residue, where Th1 cells went into aberrant mitosis. Thus, an immediate early gene product, c-Fos/AP-1, directly transactivates the wee1 kinase gene at G(1)/S. The transient increase in c-fos and wee1 kinase genes is likely to be responsible for preventing premature mitosis while the cells remain in the G(1)/S phase of the cell cycle.

Objectives. Fibronectin containing the EDA region (EDA + Fn), a molecule important for rheumatoid joint destruction, was measured in relation to the progression of joint destruction in rheumatoid arthritis (RA). Methods. Total Fn and EDA + Fn were measured by ELISA, and the concentrations of Fn in plasma and synovial fluid were compared prospectively for 2 yr with the progression of joint destruction in 41 knee joints of 37 patients with RA. The extent of joint destruction was assessed by the Larsen score and joint space narrowing in X-ray films taken before and 2 yr after measurement of EDA + Fn. Results. The concentration of synovial fluid EDA Fn showed a positive correlation with the progression of joint destruction in RA (r = 0.78). While total Fn in synovial fluid also showed a correlation with joint destruction (r = 0.54), total Fn and EDA + Fn in plasma showed no correlation with joint destruction. The concentration of synovial fluid EDA + Fn was significantly higher in patients who underwent joint replacement after the measurement of EDA Fn than in those who did not receive surgery (P < 0.029). Conclusion. Synovial fluid EDA + Fn can be a predictor of subsequent joint destruction in RA.

We have searched the human genome for genes that predispose to rheumatoid arthritis using microsatellite marker analysis and affected sib-pair linkage studies. Three principal chromosome regions of linkage, D1S253/214, D8S556 and DXS 1232/984, have been assigned for rheumatoid arthritis disease loci. We are now assigning the truncated form of Dbl proto-oncogene, which does not contain the 23rd and 24th exons, as a candidate gene for DXS 1232/984. Dbl proto-oncogene is one of the GDP/GTP exchange factors and activates Rho family GTPase. In this minireview, We describe our results and Rho family dependent signal transduction correlated with congenital diseases. Genetic linkage mapping and progress of genomic sequence-project will make it possible to identify new genes correlated with many diseases.

Human 25-hydroxyvitamin D3 (25(OH)D3) 24-hydroxylase (CYP24) cDNA was expressed in Escherichia coli, and its enzymatic and spectral properties were revealed. The reconstituted system containing the membrane fraction prepared from recombinant E. coli cells, adrenodoxin and adrenodoxin reductase was examined for the metabolism of 25(OH)D3, 1alpha,25(OH)2D3 and their related compounds. Human CYP24 demonstrated a remarkable metabolism consisting of both C-23 and C-24 hydroxylation pathways towards both 25(OH)D3 and 1alpha,25(OH)2D3, whereas rat CYP24 showed almost no C-23 hydroxylation pathway [Sakaki, T. Sawada, N. Nonaka, Y. Ohyama, Y. & Inouye, K. (1999) Eur. J. Biochem. 262, 43-48]. HPLC analysis and mass spectrometric analysis revealed that human CYP24 catalyzed all the steps of the C-23 hydroxylation pathway from 25(OH)D3 via 23S, 25(OH)2D3, 23S,25,26(OH)3D3 and 25(OH)D3-26,23-lactol to 25(OH)D3-26, 23-lactone in addition to the C-24 hydroxylation pathway from 25(OH)D3 via 24R,25(OH)2D3, 24-oxo-25(OH)D3, 24-oxo-23S,25(OH)2D3 to 24,25,26,27-tetranor-23(OH)D3. On 1alpha,25(OH)2D3 metabolism, similar results were observed. These results strongly suggest that the single enzyme human CYP24 is greatly responsible for the metabolism of both 25(OH)D3 and 1alpha,25(OH)2D3. We also succeeded in the coexpression of CYP24, adrenodoxin and NADPH-adrenodoxin reductase in E. coli. Addition of 25(OH)D3 to the recombinant E. coli cell culture yielded most of the metabolites in both the C-23 and C-24 hydroxylation pathways. Thus, the E. coli expression system for human CYP24 appears quite useful in predicting the metabolism of vitamin D analogs used as drugs.

Interferon (IFN)-alpha is the main IFN produced in response to viral infection. Low levels of IFNalpha can be detected in nasal secretions after exposure to viruses in vivo. Radioimmunoassay has shown that endogenous IFNalpha is low in children, reaches a peak in young adults, and gradually declines with aging. Importantly, this endogenous IFNalpha is significantly decreased in patients with Sjögren's syndrome (SS). IFNalpha has been tested as a therapeutic agent in patients with SS. Intramuscular human leucocyte IFNalpha increases saliva production significantly in patients with SS. Improvements have been noted in lacrimal function and in dryness symptoms. Since IFNalpha infrequently induces autoimmune phenomena and high dose IFNalpha treatment sometimes has a serious adverse event profile, treatment focus has shifted to use of low dose orally-administered IFNalpha. In a single-masked controlled trial, 60 patients with SS randomly received natural human IFNalpha 150IU 3 times a day in an oral lozenge formulation or sucralfate as control for 6 months. At study end, 15 (50%) of the 30 IFNalpha-treated patients had saliva production increases at least 100% above baseline. IFNalpha treatment was well tolerated and no patients withdrew. Labial minor salivary gland biopsies indicated significant decreases in lymphocytic infiltration accompanied by a significant increase in intact salivary gland tissue after 6 months of treatment. In another 12-week double-masked, randomised, placebo-controlled trial, stimulated saliva production in patients with SS receiving IFNalpha lozenges 150IU 3 times daily was significantly increased. This dosage was also suggestive of benefit for 5 of 7 subjective measures of oral and ocular comfort. The tolerability profile of these low dose oral IFNalpha lozenges is excellent; no serious adverse events have been recorded. Adverse effects were generally mild and there were no clinically significant changes in laboratory or clinical safety measures. Low oral doses of natural human IFNalpha thus appear to improve secretory function and relieve dryness in patients with SS without causing significant adverse events. Endogenous or orally administered IFNalpha may activate oropharyngeal lymphoid and epithelial cells and induce production of potent soluble factors which could mediate immunological reactivity. It has been suggested that IFNalpha/beta potentiates clonal expansion and survival of CD8 T cells. Stimulating effects have also been demonstrated on natural killer cell activity, which has been shown to be depressed in patients with SS. It is likely that some combination of these immunological effects results in anti-inflammatory activity and ameliorates signs and symptoms of SS.

十二指腸乳頭部腺腫は比較的稀な疾患とされている.今回われわれは,無症状で発見された十二指腸乳頭部腺腫の1例を経験したので報告する.症例は75歳,女性.検診の上部消化管内視鏡検査時に十二指腸乳頭部の腫大を指摘され,精査目的に紹介入院となった.十二指腸内視鏡検査では乳頭部に約1cm大の粗結節状隆起を認め,生検にて腺腫と診断し,十二指腸乳頭全切除,乳頭形成術を施行した.全割標本では腺腫と診断され,悪性像は認められなかった.術後約1年経過した現在も再発の徴候はない.本疾患に対しては,腺腫の遺残がなければQOLを重視した乳頭全切除術が有効な術式と考えられた.