Objectives: We aimed to clarify the differences in clinical and histological characteristics between two decompression techniques for obstructive colorectal cancer (OCRC): trans-anal decompression tube (TADT) and self-expandable metallic stent (SEMS). Methods: Overall, 96 of 175 patients with OCRC treated in our department between 2000 and 2020, were assigned into TADT (n = 44) and SEMS (n = 52) groups, and the success rate and complications of decompression, surgical outcomes, clinicopathological findings, and prognosis were compared. Results: The technical success rate was high in all patients. During surgery, the SEMS group showed significant differences in laparoscopy (p < 0.01), D3 dissection (p < 0.001), and the number of lymph nodes dissected (p < 0.01). In multivariate analysis, lymphatic invasion was associated with decompression (odds ratio [OR], 5.49; 95% confidence interval [CI], 2.12-14.20; p < 0.01) and lymph node metastasis (OR, 4.44; 95% CI, 1.69-11.70; p < 0.01), while SEMS was associated with venous invasion (OR, 0.40; 95% CI, 0.16-0.98; p = 0.04). The 5-year and recurrence-free survival rates for 70 patients with stage II and III OCRC were 76.7% and 73.2%, respectively. Although there was no significant difference in the overall survival, the recurrence-free survival was significantly better in the SEMS group (85.0% vs. 58.3%, p = 0.04). Conclusions: The results of this study suggest that SEMS may be preferable to TADT from a histological perspective.

There are certain criteria for selecting living kidney donors. However, the association between clinical characteristics of these criteria and kidney biopsy findings in living kidney donors have not yet been elucidated. Thus, we investigated the association between kidney biopsy findings and clinical characteristics defined in the Japanese guidelines for living kidney donors. A retrospective multicentre study was conducted on donors and their recipients who underwent kidney transplantation between July 2014 and June 2017. Multiple linear regression analysis and multiple logistic regression analysis were performed to investigate the association between biopsy findings and clinical characteristics. A total of 240 donors and 240 recipients were included. Age was significantly correlated with global glomerulosclerosis and intimal thickening in multiple linear regression analysis and multiple logistic regression analysis, whereas diabetes was correlated with tubular atrophy in multiple linear regression analysis after multiple imputation and multiple logistic regression analysis. Amongst the clinical factors investigated in our study, age was positively correlated and diabetes was possibly correlated with kidney tissue injury in living kidney donors. Age and diabetes may be more important for selecting suitable living kidney donors than other clinical factors.

Background Although recently strengthened immunosuppression protocols have decreased the incidence of clinical acute rejection of renal transplants, subclinical acute rejection and borderline changes remain problematic. This study was performed to evaluate the effects of antirejection therapies for early subclinical acute rejection and borderline changes. Methods In total, 269 renal transplant patients who received 3-month and 1-year protocol biopsies after renal transplantation were enrolled this study and divided into those with normal findings (Group A) and those with ≥ borderline changes (Group B) according to the 3-month pathological results. Pathological changes, graft function, and graft survival were evaluated at 1 year. Results The 3-month protocol biopsy revealed normal findings in 166 patients (Group A) and borderline changes and subclinical acute rejection in 103 patients (Group B). In Group A, 65.1% ( n = 108) of the patients maintained normal findings at 1 year, while 30.1% ( n = 50) deteriorated to ≥ borderline changes. In Group B, 52.4% ( n = 54) of patients improved to normal. Among patients with subclinical acute rejection, 25.0% ( n = 5) maintained subclinical acute rejection at 1 year despite antirejection therapy. The mean estimated glomerular filtration rate decreased from 60.4 ± 24.5 to 58.3 ± 19.0 mL/min/1.73 m 2 in Group A and from 57.2 ± 28.2 to 53.7 ± 20.3 mL/min/1.73 m 2 in Group B ( p = 0.417). The 3-, 5-, and 7-year graft survival rates were 99.4%, 99.4%, and 97.6% in Group A and 100.0%, 98.6%, and 98.6% in Group B, respectively ( p = 0.709). Conclusions Subclinical acute rejection is likely to recur. However, intervention for subclinical acute rejection in the early period after transplantation may help to prevent subsequent histological changes.

As a candidate microRNA antifibrotic effector in skin wounds, miR-146b-5p was upregulated by basic FGF, and PDGFRα was identified as a direct target of miR-146b-5p in fibroblasts. The treatment of fibroblasts with a miR-146b-5p mimic markedly downregulated the expression of PDGFRα and collagen type I. miR-146b-5p mimic transfection in wounds markedly attenuated cutaneous fibrosis, whereas a miR-146b-5p inhibitor strongly promoted fibrosis, with increases in PDGFRα and collagen I levels. These results indicate the positive effects of miR-146b-5p for the suppression of fibrosis, possibly through the inhibition of PDGFRα. The miR-146b-5p inhibitor markedly increased CD34+ vessel numbers and CD34 expression in wounds. We found miR-146b-5p+ cells in close contact with S100+ adipocytes. Moreover, we discovered the specific colocalization of the exosome marker CD81 and miR-146b-5p in the adipose tissue cells of mimic-transfected wounds, with miR-146b-5p signals being detected in the FSP1+ fibroblastic cells of adipose tissues. Therefore, fibroblastic cells of adipose tissues, which may specifically pick up and contain miR-146b-5p by exosome after transfection, may play an important role in the suppression of fibrosis. In this process, the inhibition of PDGFRα in adipose tissue cells by miR-146b-5p may lead to the loss of their PDGFRα-induced profibrotic activities, thereby suppressing fibrosis.

Background: Renin-angiotensin-aldosterone system blockers are known to reduce hypertrophy of vascular smooth muscle cells (SMCs) in hypertensive cases. However, we have reported marked proliferative changes of renal afferent arteriolar SMCs in rats induced by a long-term administration of angiotensin II type 1 receptor blockers (ARBs) and an angiotensin-converting enzyme inhibitor (ACEI). In this study, we examined the morphological changes of afferent arteriolar walls in human kidneys with or without ARBs/ACEIs. Methods: Forty-four wedge resections were taken from patients aged 45–74 years from 92 nephrectomized kidneys due to malignancy at Toho University Omori Medical Center between 2013 and 2016. They were divided into the following three groups: 18 hypertensive patients treated with antihypertensive agents including ARBs or ACEIs (the HTARB group), 6 hypertensive patients treated with calcium channel blockers without ARBs/ACEIs (the HTCCB group), and 20 normotensive patients (the normotensive group) as a control. Cases expecting vascular changes such as diabetes were excluded. In each case renal arterioles were measured as the ratio of inner/outer arteriolar diameter, and pathologists estimated morphological abnormal changes, scoring each specimen independently. Results: The ratio in the HTARB group was 0.39 ± 0.05 (mean ± SD), and was significantly the lowest among the three groups (0.46 ± 0.02 in the HTCCB, 0.53 ± 0.02 in the normotensive group; p = 0.0107 vs. HTCCB, p = 0.00001 vs. normotensive). The ratio in the three groups significantly correlated with the estimated glomerular filtration rate (r = 0.4915, p < 0.0007). The afferent arteriolar SMCs in the HTARB group frequently showed marked proliferative and irregular changes. The score of SMC abnormalities estimated regarding the proliferation, irregularity of the arrangement, and size in hilar afferent arteriolar SMCs was highest in the HTARB group and showed statistical significance (p = 0.0088, p = 0.00001, and p = 0.025 versus other two groups). Conclusions: We consider that these morphological changes in arterioles are induced by ARBs/ACEIs. These changes could induce an important suppression of glomerular hyperfiltration and could lead to glomerular ischemia. However, the clinical consequences of these morphological changes in correlation with ARBs/ACEIs were not sufficiently clear and require further analysis. We should consider renal arteriolar morphological changes when using ARBs/ACEIs.

Background: Treatment for interstitial pneumonia (IP) associated with collagen diseases has not been established. There is a need to elucidate the pathogenesis of IP and develop a novel therapy. We aimed to clarify the role of chemokine (C-X3-C motif) ligand 1 (CX3CL1, also known as fractalkine) in IP. Methods: Bleomycin (BLM) was intratracheally administered to C57BL/6 mice to induce IP. For treatment with control Ab or anti-CX3CL1 mAb, the mice were administered either Ab three times per week for 2 weeks from the day of BLM administration until euthanasia. Expressions of CX3CL1 and its unique receptor CX3CR1 in the lung tissue were examined by immunohistochemical analysis. Cellular infiltration and lung fibrosis were evaluated based on hematoxylin-eosin-staining and Sirius red staining of the lung tissue sections, respectively. Bronchoalveolar lavage fluid (BALF) cells were analyzed by flow cytometry. Results: CX3CL1 and CX3CR1 were strongly expressed in the lung tissue from mice with BLM-induced IP (BLM-IP). Treatment with anti-CX3CL1 mAb did not significantly alter inflammatory cell infiltration or fibrosis in the lung tissue. However, the number of M1-like macrophages in BALF was decreased and surface CD3 expression on T cells was increased by anti-CX3CL1 mAb treatment. Conclusions: Inhibition of CX3CL1 decreased inflammatory cells and may attenuate T cell activation in BALF. CX3CL1 inhibitor may have the potential to suppress the infiltration and activation of immune cells in IP.

Background The incidence of chronic rhinosinusitis with nasal polyps has recently increased in Japan and other East Asian countries, and this disease is called eosinophilic chronic sinusitis (ECRS) in Japan. ECRS usually occurs in adults and is frequently accompanied by refractory bronchial asthma. However, its occurrence in children under 10 years of age is rare. Here, we present an unusual case of ECRS complicated by intractable asthma in an 8-year-old boy. Case presentation Oral administration of prednisolone (10 mg/day) initially relieved the ECRS and bronchial asthma, but both returned during prednisolone dose reduction. Because nasal cavity-opening surgery was ineffective, oral administration prednisolone at 10 mg/day was continued. Pancytopenia was observed 16 months after the start of treatment, and the patient was admitted to our hospital. He was diagnosed with refractory cytopenia in childhood, but gradually improved after cyclosporine treatment. Although the dose of cyclosporine was therapeutic for asthma, it did not alleviate the asthma attacks, and the patient’s quality of life markedly decreased. We administered omalizumab even though its use was contraindicated by negative results in an inhalable antigen test. After the third administration of omalizumab, the asthma was better controlled and respiratory function improved; however, the nasal symptoms of ECRS persisted. Attempts to relieve these symptoms by increasing the therapeutic dose of omalizumab were only partially successful. We replaced omalizumab with mepolizumab; doing so slightly improved the sinusitis symptoms, but quality of life remained unsatisfactory. We repeated the nasal cavity-opening surgery. After surgery, the asthma and sinusitis were unchanged. Conclusions Omalizumab effectively treated the severe combined asthma in a young patient, but its effect on sinusitis was insufficient. More cases and long-term follow-up data are needed to better evaluate the effectiveness of mepolizumab for treatment of ECRS.

Long non‑coding RNAs (lncRNAs) have been reported to be involved in the physiological and pathological processes of tumor pathogenesis, including epithelial‑mesenchymal transition (EMT). However, epidermal growth factor receptor (EGFR)‑tyrosine kinase inhibitor (TKI) resistance is a major challenge in the treatment of advanced and recurrent EGFR‑mutant lung adenocarcinoma. An increased understanding of the underlying mechanisms would aid in the development of effective therapeutic strategies against EGFR‑TKI resistance, strategies which are urgently required for clinical therapy. In this study, long non‑coding RNA LINC00460 was identified as a novel marker of a poor response to EGFR‑TKI and prognosis. In lung cancer cells, LINC00460 promoted EGFR‑TKI resistance as a competitive decoy for miR‑149‑5p, thereby facilitating interleukin (IL)‑6 expression and inducing EMT‑like phenotypes. The knockdown or knockout of LINC00460 in gefitinib‑resistant non‑small cell lung cancer cells restored the response to EGFR‑TKI. In addition, as compared with patients with a low LINC00460 expression in tumors, those with a high LINC00460 expression had a significantly shorter progression‑free survival following gefitinib therapy, and a shorter overall survival. Therefore, LINC00460 may be a predictor of and potential therapeutic target for EGFR‑TKI resistance.

e20529 Background: Long noncoding RNA (lncRNA) is longer than 200 nucleotides and does not encode proteins. The IncRNA LINC00460 was reported to be involved in physiological and pathological processes such as proliferation, invasion, metastasis, and epithelial–mesenchymal transition (EMT). However, its role in epidermal growth factor receptor ( EGFR)-mutant lung adenocarcinoma is unclear. Methods: Non–small cell lung cancer (NSCLC) cells with knockout of LINC00460 were generated with a CRISPR-Cas9 system and used to investigate the effect of LINC00460 on gefitinib-induced cell death in an MTT assay. Western blot was used to observe change in EMT-related proteins, and changes in expression of EMT-related proteins were detected with an immunofluorescence assay. In addition, among 38 EGFR mutant lung adenocarcinomas treated by EGFR-TKI, cell-free tumor RNA extracted from blood plasma samples was analyzed for LINC00460 by using a real-time PCR assay. In addition, we assessed the association between LINC00460 expression and progression-free survival (PFS) after EGFR-TKI therapy. Results: LINC00460 was up-regulated by EGFR activation. LINC00460 expressions were significantly greater in NSCLC tumors than in adjacent normal tissues (p < 0.01). H1299 cells with LINC00460 knockout were more sensitive to gefitinib. LINC00460 was induced by transcription factors such as FOS and NF-κB in EGFR-mutant lung adenocarcinoma and gefitinib-resistant cells. LINC00460 overexpression was involved in gefitinib resistance. The MTT assay showed that LINC00460 expression was greater in cells with acquired resistance to EGFR-TKIs (gefitinib, erlotinib, and osimertinib). Western blot and immunofluorescence revealed that LINC00460 expression was associated with changes in the expression of EMT-related proteins. Patients with high LINC00460 expression in plasma cell-free tumor RNA had significantly shorter PFS after EGFR-TKI therapy than did those with low expression (median PFS: 688 vs 1089 days, respectively; p = 0.024). Conclusions: LINC00460 appears to be a novel factor involved in malignant transformation of EGFR mutant–positive lung adenocarcinoma and increased EGFR-TKI resistance. LINC00460 expression in plasma cell-free tumor RNA in EGFR-mutant adenocarcinoma was associated with shorter PFS after EGFR-TKI therapy.

Background Pathological findings of interstitial pneumonia (IP) reveal the accumulation of inflammatory cells and proliferation of fibroblasts in lung tissue. Although a treatment has not yet been established for IP, particularly for IP with collagen diseases, chemokines may play a role in the pathogenesis of IP for inflammatory cell infiltration. We previously reported that chemokine (C-X3-C motif) ligand 1 (CX3CL1, also known as fractalkine) has potential as a therapeutic target for rheumatoid arthritis (RA).1,2,3Humanized anti-human CX3CL1 monoclonal antibody (mAb) is currently undergoing clinical trials for RA.4 Objectives In the present study, we examined the therapeutic effects of CX3CL1 blockade in a murine model of IP. Methods Bleomycin (BLM)-induced IP was developed by the intratracheal administration of BLM to C57BL/6 mice. The murine lung was stained with hematoxylin and eosin, and the expression of CX3CL1 and CX3CR1, a receptor for CX3CL1, was analyzed by immunohistochemistry. Mice were treated with anti-CX3CL1 mAb for 2 weeks. Collagen eluted from the lung was quantified using the SircolTM Collagen Assay. The expression of CX3CL1 and CX3CR1 by mouse lung fibroblasts (MLFs) was examined with quantitative RT-PCR and Western blotting, respectively. Cell movement was investigated using the scrape motility assay. Results The expression of CX3CL1 and CX3CR1 was upregulated in BLM-induced IP. The treatment with anti-CX3CL1 mAb did not significantly alter inflammatory cell infiltration. However, collagen in the lung was decreased by the treatment with anti-CX3CL1 mAb. Stimulation with CX3CL1 did not alter the in vitro production of collagen by MLFs, but significantly enhanced cell movement. Conclusion CX3CL1 may be involved in increasing collagen in IP and the cell movement of MLFs. The present results suggest that CX3CL1 plays an important role in fibrosis in IP. References [1] Nanki T, et al. Inhibition of fractalkine ameliorates murine collagen-induced arthritis. J Immunol. 2004;173(11):7010-6. [2] Nanki T, et al. Migration of CX3CR1-positive T cells producing type 1 cytokines and cytotoxic molecules into the synovium of patients with rheumatoid arthritis. Arthritis Rheum. 2002;46(11):2878-83. [3] Nanki T, et al. Fractalkine/CX3CL1 in rheumatoid arthritis. Mod Rheumatol. 2017 May;27(3):392-397. [4] Tanaka Y, et al. Safety, pharmacokinetics, and efficacy of E6011, an antifractalkine monoclonal antibody, in a first-in-patient phase 1/2 study on rheumatoid arthritis. Modern rheumatology. 2018;28(1):58-65. Disclosure of Interests Soichi Yamada: None declared, Shion Miyoshi: None declared, Natsuko Kusunoki: None declared, Hiroshi Sato: None declared, Yoshikazu Kuboi Shareholder of: Eisai Co., Ltd., Employee of: KAN Research Institute, Inc. (subsidiary of Eisai Co., Ltd.), Kana Hoshino-Negishi Shareholder of: Eisai Co., Ltd., Employee of: KAN Research Institute, Inc. (subsidiary of Eisai Co., Ltd.), NAOTO ISHII Shareholder of: Eisai Co., Ltd., Employee of: KAN Research Institute, Inc. (subsidiary of Eisai Co., Ltd.), Toshio Imai Shareholder of: Eisai Co., Ltd., Employee of: KAN Research Institute, Inc. (subsidiary of Eisai Co., Ltd.), Tetsuo Mikami Speakers bureau: EA Pharma, Hiroyasu Nakano: None declared, Shinichi Kawai Grant/research support from: Nippon Zoki, Chugai, Ono, Ayumi, Hisamitu, Eisai, Japan Tobacco, Nippon Kayaku, Daiichi Sankyo, Astellas, Yutoku, and Mitsubishi Tanabe, Consultant for: Santen, Japan Tobacco, Speakers bureau: Ayumi, Chugai, Ono, Astellas, Pfizer, Celltrion, Toshihiro Nanki Grant/research support from: Chugai, Eisai, Takeda, Teijin, Eli Lilly, Bristol-Myers, AbbVie, Ono, Novartis, Asahikasei, Mitsubishi-Tanabe, Astellas, Ayumi, Pfizer, Daiichi Sankyo, Shionogi, Sanofi, Nippon Kayaku, Yutoku, Actelion, UCB, Bayer, Nihon Pharmaceutical., Consultant for: UCB, Eisai, Chugai, Ono, Gilead., Speakers bureau: Mitsubishi-Tanabe, Chugai, Eisai, Takeda, Astellas, Janssen, Eli Lilly, Ayumi, Pfizer, Asahikasei, Sanofi, Daiichi Sankyo, Otsuka, AbbVie, Ono, Teijin, Nippon Kayaku, UCB.

BackgroundSurgical management of malignant bowel obstruction carries with high morbidity and mortality. Placement of a trans-anal decompression tube (TDT) has traditionally been used for malignant bowel obstruction as a bridge to surgery. Recently, colonic metallic stent (CMS) as a bridge to surgery for malignant bowel obstruction, particularly left-sided malignant large bowel obstruction (LMLBO) caused by colorectal cancer, has been reported to be both a safe and feasible option. The aim of this retrospective study is to evaluate the clinical effects of CMS for LMLBO as a bridge to surgery compared to TDT.MethodsBetween January 2000 and December 2015, we retrospectively evaluated outcomes of 59 patients with LMLBO. We compared the outcomes of 26 patients with CMS for LMLBO between 2013 and 2015 (CMS group) with those of 33 patients managed with TDT between 2003 and 2011 (TDT group) by the historical study. LMLBO was defined as a large bowel obstruction due to a colorectal cancer that was diagnosed by computed tomography and required emergent decompression.ResultsAll patients in the CMS group were successfully decompressed (p=0.03) and could initiate oral intake after the procedure (p<0.01). Outcomes in the CMS group were superior to the TDT group in the following areas: duration of tube placement (p<0.01), surgical approach (p<0.01), operation time (p<0.01), number of resected lymph nodes (p<0.001), and rate of curative resection (p<0.01). However, no significant differences were found in the overall postoperative complication rate (p=0.151), surgical site infection rate (p=0.685), hospital length of stay (p=0.502), and the need for permanent ostomy (p=0.745). The 3-year overall survival rate of patients in the CMS and TDT groups was 73.0% and 80.9%, respectively, and this was not significant (p=0.423).ConclusionsTreatment with CMS for patients with LMLBO as a bridge to surgery is safe and demonstrated higher rates of resumption of solid food intake and temporary discharge prior to elective surgery compared to TDT. Oncological outcomes during mid-term were equivalent.

Background: A delicate balance between cell death and keratinocyte proliferation is crucial for normal skin development. Previous studies have reported that cellular FLICE (FADD-like ICE)-inhibitory protein plays a crucial role in prevention of keratinocytes from TNF-alpha-dependent apoptosis and blocking of dermatitis. However, a role for cellular FLICE-inhibitory protein in TNF-alpha-independent cell death remains unclear. Objective: We investigated contribution of TNF-alpha-dependent and TNF-alpha-independent signals to the development of dermatitis in epidermis-specific Cflar-deficient (Cflar(E-KO)) mice. Methods: We examined the histology and expression of epidermal differentiation markers and inflammatory cytokines in the skin of Cflar(E-KO); Tnfrsf1a(1/2) and Cflar(E-KO); Tnfrsf1a(-/-) mice. Mice were treated with neutralizing antibodies against Fas ligand and TNF-related apoptosis-inducing ligand to block TNF-alpha-independent cell death of Cflar(E-KO); Tnfrsf1a(-/-) mice. Results: Cflar(E-KO); Tnfrsf1a(-/-) mice were born but experienced severe dermatitis and succumbed soon after birth. Cflar(E-KO); Tnfrsf1a(+/-) mice exhibited embryonic lethality caused by massive keratinocyte apoptosis. Although keratinocytes from Cflar(E-KO); Tnfrsf1a(-/-) mice still died of apoptosis, neutralizing antibodies against Fas ligand and TNF-related apoptosis-inducing ligand substantially prolonged survival of Cflar(E-KO); Tnfrsf1a(-/-) mice. Expression of inflammatory cytokines, such as Il6 and Il17a was increased; conversely, expression of epidermal differentiation markers was severely downregulated in the skin of Cflar(E-KO); Tnfrsf1a(-/-) mice. Treatment of primary keratinocytes with IL-6 and, to a lesser extent, IL-17A suppressed expression of epidermal differentiation markers. Conclusion: TNF receptor superfamily 1 (TNFR1)-dependent or TNFR1-independent apoptosis of keratinocytes promotes inflammatory cytokine production, which subsequently blocks epidermal differentiation. Thus blockade of both TNFR1-dependent and TNFR1-independent cell death might be an alternative strategy to treat skin diseases when treatment with anti-TNF-alpha antibody alone is not sufficient.

Background: Phospholipase A2 receptor (PLA2R) is thought to be an intrinsic antigen of idiopathic membranous nephropathy (IMN), and has been widely used for the differentiation from secondary membranous nephropathies. However, the positive expression of PLA2R in the patients with hepatitis B virus associated membranous nephropathy (HBV-MN) is controversial in Asian countries, because co-localization of PLA2R and HBV antigens in glomeruli have been reported. Case Presentation: We report a case of anti PLA2R antibody positive HBV-MN that was remitted with entecavir after a relapse during treatment with lamivudine. In a renal biopsy of the case, we could confirm the co-localized glomerular deposition of HBV surface antigen (HBs-Ag) and PLA2R using double staining of immunofluorescence. We also could observe the relapse of nephrotic syndrome correlated with the increased titer of HBs-Ag, and the remission with the decrease of HBs-Ag by the change of antiviral agents. Conclusions: Our case demonstrated that the renal manifestation of HBV-MN clearly responded to antiviral agents. Furthermore, the co-localized glomerular depositions of PLA2R and HBs-Ag in HBV-MN may be concerned with the etiology of MN patients with chronic HBV infection.

PURPOSE:Paget disease of the breast is a rare cancer that originates from the nipple-areolar complex. It is often overlooked and misdiagnosed as benign chronic eczema of the nipple. We aimed to retrospectively verify whether blood flow analysis using Doppler sonography was useful for detecting the presence of Paget disease. METHODS:In this retrospective study, 12 patients with pathologically proven unilateral nipple eczematous lesions (seven with Paget disease and five with simple dermatitis) were included. Nipple blood flow signal was observed using Doppler sonography, and the detected blood flow signals were quantified using digitally recorded images. Quantified blood flow ratio and pathologically examined capillary density were evaluated between affected and unaffected nipples. Findings of mammography, grayscale sonography, and contrast-enhanced magnetic resonance imaging (CE-MRI) were reviewed. RESULTS:In patients with Paget disease, Doppler effects in the affected nipple were more clearly visible than those in the unaffected nipple. These effects were sufficiently visible to identify Paget disease. No obvious effects were observed in the affected and unaffected nipples of simple dermatitis. The quantified blood flow ratio and pathologically examined capillary density were significantly higher for the Paget lesion than those for the non-Paget lesion. The sensitivity of CE-MRI and Doppler sonography was markedly correlated, revealing blood flow changes in the nipple lesions of Paget disease. CONCLUSION:Doppler sonography visualized the proliferation of blood vessels in Paget lesions. The visualization of increased nipple blood flow using Doppler sonography is a simple and low-cost method that provides useful data for identifying Paget disease during routine medical care.

Tissue-resident macrophages and bone marrow (BM)-derived monocytes play a crucial role in the maintenance of tissue homeostasis; however, their contribution to recovery from acute tissue injury is not fully understood. To address this issue, we generated an acute murine liver injury model using hepatocyte-specific Cflar-deficient (Cflar(Hep-low)) mice. Cellular FLICE-inhibitory protein expression was down-regulated in Cflar-deficient hepatocytes, which thereby increased susceptibility of hepatocytes to death receptor-induced apoptosis. Cflar(Hep-low) mice developed acute hepatitis and recovered with clearance of apoptotic hepatocytes at 24 hours after injection of low doses of tumor necrosis factor a (TNF alpha), which could not induce hepatitis in wild-type (WT) mice. Depletion of Kupffer cells (KCs) by clodronate liposomes did not impair clearance of dying hepatocytes or exacerbate hepatitis in Cflar(Hep-low) mice. To elucidate the roles of BM-derived monocytes and neutrophils in clearance of apoptotic hepatocytes, we examined the effect of depletion of these cells on TNF alpha-induced hepatitis in Cflar(Hep-low) mice. We reconstituted Cflar(Hep-low) mice with BM cells from transgenic mice in which human diphtheria toxin receptor (DTR) was expressed under control of the lysozyme M (LysM) promoter. TNF alpha-induced infiltration of myeloid cells, including monocytes and neutrophils, was completely ablated in LysM-DTR BM-reconstituted Cflar(Hep-low) mice pretreated with diphtheria toxin, whereas KCs remained present in the livers. Under these experimental conditions, LysM-DTR BM-reconstituted Cflar(Hep-low) mice rapidly developed severe hepatitis and succumbed within several hours of TNF alpha injection. We found that serum interleukin-6 (IL-6), TNF alpha, and histone H3 were aberrantly increased in LysM-DTR BM-reconstituted, but not in WT BM-reconstituted, Cflar(Hep-low) mice following TNF alpha injection. Conclusion: These findings indicate an unexpected role of myeloid cells in decreasing serum IL-6, TNF alpha, and histone H3 levels via the suppression of TNF alpha-induced hepatocyte apoptosis.

Endogenous neural stem/progenitor cells (NPCs) can migrate toward sites of injury, but the migration activity of NPCs is insufficient to regenerate damaged brain tissue. In this study, we showed that p38 MAP kinase (p38) is expressed in doublecortin-positive adult NPCs. Experiments using the p38 inhibitor SB203580 revealed that endogenous p38 participates in NPC migration. To enhance NPC migration, we generated a cell-permeable wild-type p38 protein (PTD-p38WT) in which the HIV protein transduction domain (PTD) was fused to the N-terminus of p38. Treatment with PTD-p38WT significantly promoted the random migration of adult NPCs without affecting cell survival or differentiation; this effect depended on the cell permeability and kinase activity of the fusion protein. These findings indicate that PTD-p38WT is a novel and useful tool for unraveling the roles of p38 and that this protein provides a reasonable approach for regenerating the injured brain by enhancing NPC migration.

e23261 Background: Programmed death-ligand 1 (PD-L1) expression on immunohistochemistry is associated with response to PD-1 blockade.However, the molecular mechanism and clinical significance of PD-L1 mRNA expression and the roles of cell apoptosis and other biomarkers have not been adequately studied in patients with epidermal growth factor receptor (EGFR)-mutant lung adenocarcinoma. Methods: We studied 31 patients with recurrent postoperative EGFR-mutant lung adenocarcinoma (exon 19 deletion in 16, L858R in 14, G719C in 1) treated with gefitinib between January 2008 and January 2016. Using KCL22 cell line for calibration, we utilized relative quantification of PD-L1 mRNA expression on PCR analysis of formalin-fixed paraffin-embedded slides of surgical specimens to determine PD-L1 mRNA expression in and around the tumor. In addition, we assessed correlations of PD-L1 mRNA expression with expressions of BIM, PUMA, HER2, MET, EGFR, and VEGFA. Results: PD-L1 mRNA expression was noted in the tumor in 11 of 31 patients (35.4%) and around the tumor in 13 of 31 patients (41.9%). Patients with positive PD-L1 mRNA expression had significantly higher MET expression and significantly lower VEGFA expression as compared with patients without PD-L1 mRNA expression (p = 0.03 and p = 0.04, respectively). HER2 and EGFR expressions were not related toPD-L1 mRNA expression status. PD-L1 mRNA expression was positively correlated with total relative expressions of BIM and PUMA (r = 0.91, p < 0.001). Patients with PD-L1mRNA expression on the tumor had significantly shorter progression-free survival (PFS) after gefitinib therapy than did those without such expression (median PFS: 283 days vs. 584 days, respectively; p = 0.05). Conclusions: PD-L1 mRNA expression in EGFR-mutant lung adenocarcinoma was associated with MET, VEGFA, BIM, and PUMA mRNA expressions and with shorter PFS after gefitinib therapy.

AIM:This pilot study assessed the association of BIM deletion polymorphism and BIM RNA isoform in patients with EGFR-positive non-small cell lung cancer (NSCLC).PATIENTS AND METHODS:The study included 33 patients with EGFR-positive NSCLC treated with gefitinib. BIM deletion polymorphism and BIM RNA isoform (EL/L/S/γ) were determined by polymerase chain reaction (PCR).RESULTS:BIM-γ expression was significantly higher in patients with BIM deletion polymorphism than among those without BIM deletion polymorphism inside tumors (p=0.038) and around tumors (p=0.0024). Relative BIM-γ expression was significantly higher in patients with BIM deletion polymorphism than among those without BIM deletion polymorphism (p=0.0017). Patients with BIM-γ had significantly shorter progression-free survival than those without BIM-γ (median: 304 vs. 732 days; p=0.023).CONCLUSION:Expression of BIM-γ mRNA and BIM deletion polymorphism were strongly associated. BIM-γ overexpression may have a role in apoptosis related to EGFR-tyrosine kinase inhibitor.