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CD34 is a transmembrane phosphoglycoprotein used to selectively enrich bone marrow in hematopoietic stem cells for transplantation. Treating rats with CD34+ cells derived from human umbilical cord blood before or after heat stroke has been shown to promote survival. We investigated whether CD34– human placenta-derived stem cells (PDMSCs) could improve survival following heat stroke in rats. Rats were subjected to heat stress (42°C for 98 min) to induce heat stroke. Intravenous administration of PDMSCs 1 day before or immediately after the onset of heat stroke improved survival by 60% and 20%, respectively. Pre-treatment with CD34− PDMSCs protected against heat stroke injury more effectively than that treatment after injury. PDMSCs treatment attenuated cerebrovascular dysfunction, the inflammatory response, and lipid peroxidation. These data suggest human PDMSCs protect against heat stroke injury in rats. Moreover, these effects do not require the presence of CD34+ cells.

// Chun-Wei Wang 1, 2 , Chiung-Chiao Huang 1 , Pei-Hsin Chou 4 , Yu-Ping Chang 1 , Shouzuo Wei 5 , Frederick Peter Guengerich 6 , Yueh-Ching Chou 3, 7, 9 , Sheng-Fan Wang 1, 7 , Ping-Shan Lai 8 , Pavel Soucek 11 and Yune-Fang Ueng 1, 2, 3, 10 1 National Research Institute of Chinese Medicine, Ministry of Health and Welfare, Taipei, Taiwan, R.O.C 2 Institute of Biopharmaceutical Sciences, School of Pharmacy, National Yang-Ming University, Taipei, Taiwan, R.O.C 3 Department of Pharmacology, School of Medicine, National Yang-Ming University, Taipei, Taiwan, R.O.C 4 Department of Environmental Engineering, National Chung-Kung University, Tainan, Taiwan, R.O.C 5 Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN, USA 6 Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN, USA 7 Department of Pharmacy, Taipei Veterans General Hospital, Taipei, Taiwan, R.O.C 8 Department of Chemistry, College of Science, National Chung-Hsin University, Taichung, Taiwan, R.O.C 9 Department of Pharmacy, Taipei Medical University, Taipei, Taiwan, R.O.C 10 Institute of Medical Sciences, Taipei Medical University, Taipei, Taiwan, R.O.C 11 Department of Toxicogenomics, National Institute of Public Health, Prague, Czech Republic Correspondence to: Yune-Fang Ueng, email: ueng@nricm.edu.tw Keywords: 7-ketocholesterol, 27-hydroxycholesterol, doxorubicin, P-glycoprotein, estrogen receptor Received: March 13, 2017 Accepted: June 27, 2017 Published: August 02, 2017 ABSTRACT Hypercholesterolemia is one of the risk factors for poor outcome in breast cancer therapy. To elucidate the influence of the main circulating oxysterols, cholesterol oxidation products, on the cell-killing effect of doxorubicin, cells were exposed to oxysterols at a subtoxic concentration. When cells were exposed to oxysterols in fetal bovine serum-supplemented medium, 7-ketocholesterol (7-KC), but not 27-hydroxycholesterol (27-HC), decreased the cytotoxicity of doxorubicin in MCF-7 (high estrogen receptor (ER)α/ERβ ratio) cells and the decreased cytotoxicity was restored by the P-glycoprotein inhibitor verapamil. 7-KC stimulated the efflux function of P-glycoprotein and reduced intracellular doxorubicin accumulation in MCF-7 but not in ERα(-) MDA-MB-231 and the resistant MCF-7/ADR cells. In MCF-7 cells, 7-KC increased the mRNA and protein levels of P-glycoprotein. The 7-KC-suppressed doxorubicin accumulation was restored by the fluvestrant and ERα knockdown. In a yeast reporter assay, the ERα activation by 7-KC was more potent than 27-HC. 7-KC, but not 27-HC, stimulated the expression of an ER target, Trefoil factor 1 in MCF-7 cells. When charcoal-stripped fetal bovine serum was used, both 7-KC and 27-HC induced Trefoil factor 1 expression and reduced doxorubicin accumulation in MCF-7 cells. 7-KC-reduced doxorubicin accumulation could be reversed by inhibitors of phosphatidylinositol 3-kinase, Akt, and mammalian target of rapamycin (mTOR). These findings demonstrate that 7-KC decreases the cytotoxicity of doxorubicin through the up-regulation of P-glycoprotein in an ERα- and mTOR-dependent pathway. The 7-KC- and 27-HC-elicited estrogenic effects are crucial in the P-glycoprotein induction in breast cancer cells.

// I-Chun Kuo 1, * , Huang-Kai Kao 2, 3, * , Yenlin Huang 4 , Chun-I Wang 1 , Jui-Shan Yi 1, 5 , Ying Liang 5 , Chun-Ta Liao 1, 3 , Tzu-Chen Yen 3, 6 , Chih-Ching Wu 1, 5, 7 and Kai-Ping Chang 1, 3, 5 1 Department of Otolaryngology - Head & Neck Surgery, Chang Gung Memorial Hospital, Kwei-Shan, Tao-Yuan, Taiwan 2 Department of Plastic & Reconstructive Surgery, Chang Gung Memorial Hospital, Kwei-Shan, Tao-Yuan, Taiwan 3 College of Medicine, Chang Gung University, Kwei-Shan, Tao-Yuan, Taiwan 4 Departments of Pathology, Chang Gung Memorial Hospital, Kwei-Shan, Tao-Yuan, Taiwan 5 Molecular Medicine Research Center, Chang Gung University, Kwei-Shan, Tao-Yuan, Taiwan 6 Departments of Nuclear Medicine, Chang Gung Memorial Hospital, Kwei-Shan, Tao-Yuan, Taiwan 7 Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Kwei-Shan, Tao-Yuan, Taiwan * These authors have contributed equally to this work Correspondence to: Kai-Ping Chang, email: dr.kpchang@gmail.com Chih-Ching Wu, email: luckywu@mail.cgu.edu.tw Keywords: OSCC, proteome, metastasis, prognosis, ERAP2 Received: September 30, 2016 Accepted: May 23, 2017 Published: June 27, 2017 ABSTRACT Oral cavity squamous cell carcinoma (OSCC) is a leading cause of cancer-related deaths worldwide and associated with poor prognosis and mortality. Discovery of proteins that can improve OSCC treatment is needed. Using comparative proteome profiling of primary cells derived from OSCC and adjacent noncancerous epithelium, endoplasmic reticulum aminopeptidases 2 (ERAP2) has been identified as an OSCC-associated protein. Compared with the adjacent normal tissues, ERAP2 levels were determined to be significantly elevated in OSCC tissues using quantitative real-time PCR and immunohistochemistry. Importantly, overexpression of ERAP2 was associated with positive N stage, advanced overall stage, positive perineural invasion, and tumor depth ( P = 0.041, 0.015, 0.010, and 0.032, respectively). The overall survival rates of patients without and with the ERAP2 overexpression were 71.9% and 56.0%, respectively ( P = 0.029). Furthermore, knockdown of ERAP2 inhibited the migration and invasion abilities of OSCC cells. Our results collectively show that ERAP2 overexpression is associated with the cervical metastasis and poorer prognosis of OSCC.

// Feng-Hua Liu 1 , Shan-Shan Li 2, , Xiao-Xi Li 2, , Shuai Wang 2, 3 , Mao-Gang Li 2 , Li Guan 4 , Tian-Gang Luan 4 , Zhi-Gang Liu 2 , Zhan-Ju Liu 1 and Ping-Chang Yang 2 1 The Department of Gastroenterology, The Shanghai Tenth People’s Hospital of Tongji University, Shanghai 200072, China 2 The Research Center of Allergy & Immunology, Shenzhen University School of Medicine, Shenzhen 518060, China 3 Longgang ENT Hospital, Shenzhen ENT Institute, Shenzhen 518116, China 4 Affiliated Luohu Hospital, Shenzhen University, Shenzhen 518001, China Correspondence to: Ping-Chang Yang, email: pcy2356@szu.edu.cn Zhan-Ju Liu, email: liuzhanju88@126.com Keywords: intestine, epithelium, barrier function, vitamin D, histone deacetylase Received: October 15, 2016 Accepted: April 12, 2017 Published: May 08, 2017 ABSTRACT Intestinal epithelial barrier dysfunction and vitamin D (VitD)-deficiency play a critical role in a large number of diseases. The histone deacetylases (HDAC) are associated with a large number of immune diseases. This study tests a hypothesis that the interaction between VitD and HDAC is associated with the regulation of epithelial barrier functions. In this study, human intestinal epithelial cell line, T84 cells, was cultured into monolayers to be used as a model to test the epithelial barrier functions. We observed that in a VitD-deficient environment, the T84 monolayer barrier function was compromised. Exposure to calcitriol (the active form of VitD3) in the culture increased the expression of VitD receptor (VDR) in T84 cells. In a VitD-sufficient environment, VDR formed a complex with histone deacetylase-11 (HDAC11); the complex was markedly decreased in a VitD-deficient environment. We also observed that significantly more binding of HDAC11 to the promoter of the tight junction proteins inhibit the gene transcription activities of these loci in the VitD-deficient environment, which were abolished by the presence of calcitriol in the culture. In conclusion, the interaction between VDR and HDAC11 plays a crucial role in the maintenance of the epithelial barrier integrity.

// Jiangping Song 1, * , Wenjun Su 1, * , Xiao Chen 1 , Qian Zhao 1 , Ningning Zhang 1 , Mao-Gang Li 2 , Ping-Chang Yang 2 , Liqing Wang 1 1 State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Xicheng District, Beijing, 100037, China 2 The Research Center of Allergy and Immunology, Shenzhen University School of Medicine, Shenzhen, 518060, China * These authors contributed equally to this work Correspondence to: Liqing Wang, email: fwsongjiangping@126.com Keywords: B lymphocyte, interleukin-10, micro RNA, heart transplantation, cortisol Received: September 20, 2016 Accepted: February 28, 2017 Published: March 08, 2017 ABSTRACT The immune tolerance to the transplant heart survival is critical. Regulatory B cells are one of the major immune regulatory cell populations in the immune tolerance. Micro RNAs (miR) can regulate the activities of immune cells, such as the expression of interleukin (IL)-10 by B cells. This study tests a hypothesis that micro RNA (miR)-98 plays a role in the regulation of interleukin (IL)-10 expression in B cells (B10 cell) after heart transplantation. In this study, the peripheral blood samples were collected from patients before and after heart transplantation. The expression of miR-98 and IL-10 in B cells was assessed by real time RT-PCR. An allograft heart transplantation mouse model was developed. We observed that after heart transplantation, the frequency of peripheral B10 cell and the IL-10 mRNA levels in peripheral B cells were significantly decreased, the levels of miR-98 were increased in peripheral B cells and the serum levels of cortisol were increased in the patients. Treating naive B cells with cortisol in the culture suppressed the expression of IL-10 in B cells, which was abolished by knocking down the miR-98 gene. Administration with anti-miR-98, or cortisol inhibitor, or adoptive transfer with B10 cells, significantly enhanced the survival rate and time of mice received allograft heart transplantation. In conclusion, the enhancement of serum cortisol affects the immune tolerant feature of B cells, which can be attenuated by anti-miR-98-carrying liposomes.

// Sheng-Chieh Chan 1,2,* , Nai-Ming Cheng 2,3,* , Chia-Hsun Hsieh 4 , Shu-Hang Ng 5,6 , Chien-Yu Lin 7 , Tzu-Chen Yen 2,3 , Cheng-Lung Hsu 4 , Hung-Ming Wan 4 , Chun-Ta Liao 8 , Kai-Ping Chang 8 and Jiun-Jie Wang 6 1 Department of Nuclear Medicine, Keelung Chang Gung Memorial Hospital, Keelung, Taiwan 2 Molecular Imaging Center, Linkou Chang Gung Memorial Hospital and Chang Gung University, Taoyuan, Taiwan 3 Department of Nuclear Medicine, Linkou Chang Gung Memorial Hospital and Chang Gung University, Taoyuan, Taiwan 4 Department of Internal Medicine, Division of Medical Oncology, Linkou Chang Gung Memorial Hospital and Chang Gung University, Taoyuan, Taiwan 5 Department of Diagnostic Radiology, Linkou Chang Gung Memorial Hospital and Chang Gung University, Taoyuan, Taiwan 6 Department of Medical Imaging and Radiological Sciences, Chang Gung University, Taoyuan, Taiwan 7 Department of Radiation Oncology, Linkou Chang Gung Memorial Hospital and Chang Gung University, Taoyuan, Taiwan 8 Department of Otorhinolaryngology, Linkou Chang Gung Memorial Hospital and Chang Gung University, Taoyuan, Taiwan * These authors have contributed equally to this work Correspondence to: Shu-Hang Ng, email: // Jiun-Jie Wang, email: // Keywords : PET, heterogeneity, DCE-MRI, DWI, head and neck cancer Received : July 22, 2016 Accepted : February 20, 2017 Published : March 04, 2017 Abstract Background: In this study, PET heterogeneity was combined with functional MRI techniques to refine the prediction of prognosis in patients with oropharyngeal or hypopharyngeal squamous cell carcinoma (OHSCC). Methods: A total of 124 patients with primary advanced OHSCC who underwent pretreatment 18 F-FDG PET/CT, dynamic contrast-enhanced MR imaging (DCE-MRI), and diffusion-weighted MR imaging (DWI) were enrolled. Conventional and heterogeneity parameters from 18 F-FDG PET as well as perfusion parameters from DCE-MRI and diffusion parameter from DWI of primary tumors were analyzed in relation to recurrence-free survival (RFS) and overall survival (OS). Results: Multivariate analysis identified hypopharyngeal tumors ( P = 0.038), alcohol drinking ( P = 0.006), K trans ≤ 0.5512 ( P = 0.017), and K ep ≤ 0.8872 ( P = 0.005) as adverse prognostic factors for RFS. Smoking ( p = 0.009), K trans ≤ 0.5512 ( P = 0.0002), K ep ≤ 0.8872 ( P = 0.004), and the PET heterogeneity parameter uniformity ≤ 0.00381 ( P = 0.028) were independent predictors of poor OS. The combination of PET uniformity with DCE-MRI parameters and smoking allowed distinguishing four prognostic groups, with 3-year OS rates of 100%, 76.6%, 57.4%, and 7.1%, respectively ( P < 0.0001). This prognostic system appeared superior to both the TNM staging system ( P = 0.186) and the combination of conventional PET parameters with DCE-MRI ( P = 0.004). Conclusions: Multiparametric imaging based on PET heterogeneity and DCE-MRI parameters combined with clinical risk factors is superior to the concomitant use of functional MRI coupled with conventional PET parameters. This approach may improve the prognostic stratification of OHSCC patients.

// Xiang-Qian Luo 1, 2, * , Jian-Bo Shao 1, 2, * , Rui-Di Xie 2 , Lu Zeng 2 , Xiao-Xi Li 2 , Shu-Qi Qiu 3 , Xiao-Rui Geng 3 , Li-Tao Yang 3, 4 , Lin-Jing Li 4 , Da-Bo Liu 1 , Zhi-Gang Liu 2 and Ping-Chang Yang 2 1 Department of Otolaryngology, Guangzhou Women and Children’s Medical Center, Guangzhou Medical University, Guangzhou 510010, China 2 The Research Center of Allergy and Immunology, Shenzhen University School of Medicine, Shenzhen 518060, China 3 Longgang ENT Hospital, Shenzhen 518116, China 4 Brain Body Institute, McMaster University, Hamilton, ON, L8N 4A6, Canada * These authors equally contributed to this work Correspondence to: Ping-Chang Yang, email: pcy2356@szu.edu.cn Zhi-Gang Liu, email: lzg@szu.edu.cn Keywords: nose, nasal polyp, dendritic cells, micro RNA, interleukin-10 Received: November 02, 2016 Accepted: March 14, 2017 Published: March 24, 2017 ABSTRACT The pathogenesis of nasal polyp is to be further investigated. Micro RNA (miR) plays a role in the development of allergic inflammation. Interleukin (IL)-10-producing dendritic cells (DC) have immune tolerogenic properties. This study test a hypothesis that miR-17-92 cluster is associated with suppressing IL-10 in peripheral DC. In this study, peripheral blood samples were obtained from 26 patients with nasal polyp. The CD11c DCs were isolated from the blood samples and analyzed for the expression of IL-10. We observed that, as compared with healthy subjects, the IL-10 expression in peripheral DC was significantly lower in polyp patients. The levels of miR-19a, but not the rest 5 members of the miR-17-92 cluster, were markedly higher in DCs in polyp group. Exposure to recombinant IL-4 suppressed the IL-10 expression in DCs, which was abolished by blocking histone deacetylase-11 or knocking down the miR-19a gene in DCs. We conclude that miR-19a plays a critical role in the suppression of IL-10 in peripheral DCs, which may be a target in the immune therapy for nasal polyp.

// Yixiang Mao 1, 2, 5 , Jianjun Shen 4 , Yue Lu 4 , Kevin Lin 4 , Huamin Wang 3 , Yanan Li 2 , Ping Chang 2 , Mary G. Walker 4 and Donghui Li 2 1 Department of Oncology, The First Affiliated Hospital of Soochow University, Suzhou 215007, China 2 Department of Gastrointestinal Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas 77030, USA 3 Department of Pathology and Department of Translational Molecular Pathology, The University of Texas MD Anderson Cancer Center, Houston, Texas 77030, USA 4 Department of Epigenetics and Molecular Carcinogenesis, The University of Texas MD Anderson Cancer Center, Smithville, Texas 78957, USA 5 Department of Medical Oncology, Fudan University Shanghai Cancer Center, Shanghai 200032, China Correspondence to: Donghui Li, email: dli@mdanderson.org Yixiang Mao, email: maoyix@suda.edu.cn Keywords: pancreatic cancer, RNA sequencing, transcriptome, pathway analysis Received: June 03, 2016 Accepted: February 27, 2017 Published: March 22, 2017 ABSTRACT Gene expression microarrays have identified many tumor markers and therapeutic targets for pancreatic ductal adenocarcinoma (PDAC). However, microarray profilings have limited sensitivity and are prone to cross-hybridization between homologous DNA fragments. Here, we perform a transcriptome analysis of paired tumor and adjacent benign pancreatic tissues from 10 patients who underwent resection for PDAC. We identify a total of 2736 differentially expressed genes (DEGs) with false discovery rate less than 0.05, including 1554 upregulated, 1182 downregulated, and 6 microRNAs (miR-614, miR-217, miR-27b, miR-4451, miR-3609, and miR-612). Overexpression of five DEGs, i.e. KRT16 , HOXA10 , CDX1 , SI , and SERPINB5 in tumors is confirmed by RT-PCR in 20 additional tissues. Overexpression of KRT16 in PDAC is also verified on protein level. In addition, top canonical pathways such as granulocyte adhesion and diapedesis pathway have been identified. Our study represents a comprehensive characterization of the PDAC transcriptome and provides insight to the mechanisms of pancreatic carcinogenesis and potential biomarkers and novel therapeutic targets for pancreatic cancer.

// Rui-Di Xie 1, 2, * , Ling-Zhi Xu 1, * , Li-Tao Yang 3, 4, * , Shuai Wang 3 , Qi Liu 2 , Zhi-Gang Liu 1 , Ping-Chang Yang 1 1 The Research Center of Allergy & Immunology, Shenzhen University School of Medicine, Shenzhen, China 2 Periodontal Department, the Affiliated Hospital of Zunyi Medical College, Zunyi, China 3 The ENT Hospital of Shenzhen University and Shenzhen ENT Institute, Shenzhen, China 4 Brain Body Institute, McMaster University, Hamilton, ON, Canada * These authors contributed equally to this work Correspondence to: Ping-Chang Yang, email: pcy2356@szu.edu.cn Zhi-Gang Liu, email: lzg@szu.edu.cn Qi Liu, email: liuqi1964@hotmail.com Keywords: oral mucosa, oral allergy, peanut, micro RNA-98, galectin-1 Received: August 25, 2016 Accepted: December 27, 2016 Published: January 10, 2017 ABSTRACT Background and aims: The pathogenesis of oral-intestinal allergy syndrome (OIAS) has not been well understood. Published data indicate that galectin (Gal) 1 has immune regulatory functions. This study tests a hypothesis that Gal1 inhibits oral-intestinal allergy syndrome. Methods: Mice were sensitized to peanut extracts (PE) via the buccal mucosa with or without using Gal1 together. Results: Upon re-exposure to specific antigen, the OIAS mice showed the systemic allergic response, the oral allergic reactions, and intestinal allergic inflammation, including increases in serum histamine, drop of the core temperature, higher levels of PE-specific IgE and interleukin (IL)-4. Increases in mast cell and eosinophil in the oral mucosa and intestinal mucosa were also observed. The OIAS was inhibited by co-administration with Gal1 via a mechanism of suppressing micro RNA (miR)-98 and reversing the expression of IL-10 in CD14+ cells in the intestine. Conclusions: The OIAS can be induced by applying specific antigens to the oral mucosa, which can be inhibited by co-administration with Gal1.

// Chang Liu 1, 4 , Cathy Eng 1 , Jianjun Shen 3 , Yue Lu 3 , Yoko Takata 3 , Amir Mehdizadeh 1 , George J. Chang 2 , Miguel A. Rodriguez-Bigas 2 , Yanan Li 1 , Ping Chang 1 , Yixiang Mao 1 , Manal M. Hassan 1 , Fangyu Wang 4 , Donghui Li 1 1 Department of Gastrointestinal Medical Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA 2 Department of Surgical Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA 3 Department of Epigenetics and Molecular Carcinogenesis, The University of Texas MD Anderson Cancer Center, Smithville, Texas, USA 4 Department of Gastroenterology and Hepatology, Jinling Hospital, Southern Medical University, Nanjing, China Correspondence to: Donghui Li, email: dli@mdanderson.org Fangyu Wang, email: wangfy65@nju.edu.cn Keywords: exosome, microRNA, colon cancer, qRT-PCR, Illumina RNA sequencing Received: April 22, 2016 Accepted: October 11, 2016 Published: October 24, 2016 ABSTRACT Purpose: The study was aimed to evaluate the prognostic or predictive value of serum exosomal microRNAs (miRNAs) for tumor recurrence and response to adjuvant therapy in stage II and stage III colon cancer. Results: 145 differentially expressed mature miRNAs were identified ( P

The therapeutic efficacy of allergen specific immunotherapy (SIT) on allergic diseases is to be improved. Probiotics can regulate immune response. This study aims to promote the effect of SIT on allergic rhinitis (AR) by co-administration with Clostridium butyricum (Cb). In this study, patients with AR sensitized to mite allergens were enrolled to this study, and treated with SIT or/and Cb. The therapeutic efficacy was evaluated by the total nasal symptom scores (NSS), medication scores, serum specific IgE levels and T helper (Th)2 cytokine levels. The improvement of immune regulation in the AR patients was assessed by immunologic approaches. The results showed that treating AR patients with SIT alone markedly reduced NSS and medication scores; but did not alter the serum specific IgE, Th2 cytokines and skin prick test (SPT) index. The clinical symptoms on AR in SIT group relapsed one month after stopping SIT. Co-administration of Cb significantly enhanced the efficacy of SIT on AR as shown by suppression of NSS, medication scores, serum specific IgE, Th2 cytokines and SPT index; the regulatory B cell frequency was also markedly increased. Such an effect on AR was maintained throughout the observation period even after stopping the treatment. Butyrate blocked the activation of histone deacetylase-1, the downstream activities of epsilon chain promoter activation, and the IgE production in the antigen specific B cells. On the other hand, butyrate induced the IL-10 expression in B cells with a premise of the B cell receptor activation by specific antigens. In conclusion, administration with Cb can markedly enhance the efficacy of SIT on AR.

// Hung-Ming Wang 1, * , Tung-Liang Lin 1, * , Yung-Chia Kuo 1 , Hsin-Pai Li 2 , Kai-Ping Chang 3 , Chien-Yu Lin 4 , Hsien-Chi Fan 1 , An-Chi Lin 1 , Chia-Hsun Hsieh 1 , Ngan-Ming Tsang 4 , Li-Yu Lee 5 , Sheng-Chieh Chan 6 , Kar-Wai Lui 7 , Yu-Sun Chang 2 , Cheng-Lung Hsu 1 1 Division of Hematology-Oncology, Department of Internal Medicine, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan, Taiwan, ROC 2 Department of Cell and Molecular Biology, Chang Gung University, Taoyuan, Taiwan, ROC 3 Department of Otolaryngology-Head and Neck Surgery, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan, Taiwan, ROC 4 Department of Radiation Oncology, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan, Taiwan, ROC 5 Department of Pathology, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan, Taiwan, ROC 6 Department of Nuclear Medicine, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan, Taiwan, ROC 7 Department of Diagnostic Radiology, Chang Gung Memorial Hospital, Chang Gung University, Taoyuan, Taiwan, ROC * These authors contributed equally to this work Correspondence to: Cheng-Lung Hsu, email: hsu2221@adm.cgmh.org.tw Keywords: nasopharyngeal carcinoma, metastasis, EB virus, EBV DNA, IP-10 Received: March 10, 2016 Accepted: June 09, 2016 Published: July 18, 2016 ABSTRACT Differential overall survival of nasopharyngeal carcinoma (NPC) with different organ site metastases has been documented. Here, we attempted to determine the underlying mechanisms by assessing plasma and tumor tissue markers in relation to patient survival. Pretreatment plasma Epstein-Barr virus (EBV) DNA concentrations, cytokines and tissue macrophages, proliferation and apoptosis markers were determined in 178 patients with metastatic NPC. The median overall survival (OS) was 19 months. Patients with single organ metastases had better outcomes than those with multiple organ metastases (median OS: 26 months vs. 16 months), with statistical significance. Among the single organ involvement cases, patients with lung metastasis only showed longer survival than those with bone or liver involvement (median OS: 50 months vs. 21 months vs. 18 months; P < 0.001). Pretreatment plasma EBV DNA concentrations were lower in patients with lung metastasis than bone or liver metastasis among single organ site groups. Plasma interferon-γ-inducible protein-10 (IP-10) and monocyte chemotactic protein-1 (MCP-1) expression levels were correlated with differential single organ site metastasis OS and EBV DNA load. Liver metastatic tissue had higher density of infiltrating macrophages and proliferative index than the lung metastatic group. Low pretreatment plasma EBV DNA load, expression of cytokines, such as IP-10 and MCP-1, tissue macrophage infiltration, and proliferative index may contribute to the differences in overall survival.

Angiopoietin-1 (Ang-1) and angiopoietin-2 (Ang-2) play critical roles in angiogenesis in hepatocellular carcinoma (HCC). In addition, recent data suggest that Ang-1/Ang-2 are involved in regulating the immune response. The aim of our study was to explore the clinical prognostic significance of plasma Ang-1 and Ang-2 in HCC. We prospectively enrolled and collected data and blood samples from 767 HCC patients treated at MD Anderson Cancer Center between 2001 and 2014. Controls consisted of cirrhotic patients (n = 75) and healthy volunteers (n = 200). The cutoff value was the median level of each angiogenic factor. Overall survival (OS) was estimated by Kaplan-Meier curves and compared by the log-rank test. Higher plasma Ang-2 was significantly associated with advanced clinicopathologic features of advanced HCC and lower OS. Median OS was 61.8 months (95% confidence interval [CI], 45.1-78.5 months) for low Ang-2 compared with 28.5 months (95% CI, 24.8-32.1 months) for high Ang-2 (p < 0.001). In contrast, higher Ang-1 was associated with longer OS. Median OS was 37.2 months (95% CI, 31.0-43.4 months) for high Ang-1 compared with 26.2 months (95% CI, 22.2-30.3 months) for those with low Ang-1 (p = 0.043). In conclusion, our findings indicate that plasma Ang-1 and Ang-2 levels are potential diagnostic and prognostic biomarkers in HCC.

Nasopharyngeal carcinoma (NPC) is an Epstein Barr virus (EBV)-related malignancy in which the tumor microenvironment plays a pivotal role in tumor progression. Here, we developed two patient-derived xenograft (PDX) mouse lines from engrafted NPC metastatic tumors. Positive staining for EBV-encoded small RNAs confirmed that these tumors harbored EBV, and gene expression profile analyses further showed that the PDX was highly similar to the primary parent tumor. In vivo drug screening using the PDX system demonstrated that gemcitabine had the best antitumor effect among the tested drugs. The donor of this PDX also showed excellent responsiveness to gemcitabine treatment. The combination of gemcitabine and valproic acid exerted synergistic antitumor effects. Further addition of ganciclovir to this two-drug combination regimen enhanced cytolytic viral activation, yielding the best antitumor response among tested regimens. Treatment with this three-drug combination regimen decreased plasma EBV-DNA load, tumor viral concentration, and the number of viable tumor cells to a greater extent than the two-drug gemcitabine and valproic acid combination. These results highlight the value of PDX models in the development of EBV-targeted strategies to treat NPC.

Epithelial cell adhesion molecule (EpCAM) is known to be overexpressed in epithelial cancers associated with enhanced malignant potential, particularly colorectal carcinoma (CRC) and head and neck squamous cell carcinoma (HNSCC). However, it is unknown whether progression of malignance can be directly inhibited by targeting EpCAM. Here, we have generated five novel monoclonal antibodies (mAbs) against EpCAM. One of these anti-EpCAM mAbs, EpAb2-6, was found to induce cancer cell apoptosis in vitro, inhibit tumor growth, and prolong the overall survival of both a pancreatic cancer metastatic mouse model and mice with human colon carcinoma xenografts. EpAb2-6 also increases the therapeutic efficacy of irinotecan, fluorouracil, and leucovorin (IFL) therapy in a colon cancer animal model and gemcitabine therapy in a pancreatic cancer animal model. Furthermore, EpAb2-6, which binds to positions Y95 and D96 of the EGF-II/TY domain of EpCAM, inhibits production of EpICD, thereby decreasing its translocation and subsequent signal activation. Collectively, our results indicate that the novel anti-EpCAM mAb can potentially be used for cancer-targeted therapy.

// Chien-Wei Lee 1 , Kai-Ping Chang 3 , Yan-Yu Chen 5 , Ying Liang 5 , Chuen Hsueh 4, 5, 6 , Jau-Song Yu 1, 2, 5 , Yu-Sun Chang 1, 5 , Chia-Jung Yu 1, 2, 5 1 Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan 2 Department of Cell and Molecular Biology, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan 3 Department of Otolaryngology-Head and Neck Surgery, Chang Gung Memorial Hospital, Lin-Kou, Tao-Yuan, Taiwan 4 Department of Pathology, Chang Gung Memorial Hospital, Lin-Kou, Tao-Yuan, Taiwan 5 Molecular Medicine Research Center, Chang Gung University, Tao-Yuan, Taiwan 6 Pathology Core, Chang Gung University, Tao-Yuan, Taiwan Correspondence to: Chia-Jung Yu, e-mail: yucj1124@mail.cgu.edu.tw Keywords: tryptophanyl-tRNA synthetase, oral squamous cell carcinoma, migration, invasion Received: February 10, 2015 Accepted: June 05, 2015 Published: June 17, 2015 ABSTRACT Oral squamous cell carcinoma (OSCC) is one of the most common neoplasms worldwide. Previously, we identified the angiostatic agent tryptophanyl-tRNA synthetase (TrpRS) as a dysregulated protein in OSCC based on a proteomics approach. Herein, we show that TrpRS is overexpressed in OSCC tissues (139/146, 95.2%) compared with adjacent normal tissues and that TrpRS expression positively correlates with tumor stage, overall TNM stage, perineural invasion and tumor depth. Importantly, the TrpRS levels were significantly higher in tumor cells from metastatic lymph nodes than in corresponding primary tumor cells. TrpRS knockdown or treatment with conditioned media obtained from TrpRS-knockdown cells significantly reduced oral cancer cell viability and invasiveness. TrpRS overexpression promoted cell migration and invasion. In addition, the extracellular addition of TrpRS rescued the invasion ability of TrpRS-knockdown cells. Subcellular fractionation and immunofluorescence staining further revealed that TrpRS was distributed on the cell surface, suggesting that secreted TrpRS promotes OSCC progression via an extrinsic pathway. Collectively, our results demonstrated the clinical significance and a novel role of TrpRS in OSCC.

// Hsin-Pai Li 1,2,3,* , Chen-Ching Peng 1,* , I-Che Chung 2 , Mei-Yuan Huang 2 , Shao-Tung Huang 1 , Chia-Chun Chen 2 , Kai-Ping Chang 4 , Cheng-Lung Hsu 5 , Yu-Sun Chang 1,2 1 Graduate Institute of Biomedical Sciences, Chang Gung University, Taoyuan, Taiwan, Republic of China (ROC) 2 Molecular Medicine Research Center, Chang Gung University, Taoyuan, Taiwan, Republic of China (ROC) 3 Department of Microbiology and Immunology Medical School, Chang Gung University, Taoyuan, Taiwan, Republic of China (ROC) 4 Department of Otolaryngology-Head and Neck Surgery, Chang Gung Memorial Hospital, Lin-Kou, Taiwan, ROC 5 Division of Hematology-Oncology, Chang Gung Memorial Hospital, Lin-Kou, Taiwan, ROC * These authors contributed equally to this work. Correspondence: Hsin-Pai Li , email: // Keywords : HOXA2, DNA methylation, NPC, MMP9 Received : September 4, 2013 Accepted : November 2, 2013 Published : November 4, 2013 Abstract Nasopharyngeal carcinoma (NPC) is notorious for its high invasiveness and metastatic ability. In this study, we identified a differential hypermethylated transcription repressor, Homeobox A2 (HOXA2) , which may render NPC cells invasive and metastatic. Aberrant hypermethylation of HOXA2 led to low RNA expression in NPC tumors and cells. Addition of methylation inhibitor 5’Aza restored HOXA2 RNA expression in NPC cells. Methylated HOXA2 promoter reduces the binding affinity of the transcriptional co-activator p300, causing transcriptionalrepression of HOXA2 . In NPC cells, re-expression of ectopic HOXA2 wascorrelated with decreased invasive ability and reduced metalloproteinase MMP-9 RNA and protein expression. Promoter, ChIP and DNA-pull down assays indicated that HOXA2 competes with the transcription activator, TATA-box binding protein (TBP) for a recognition sequence near the MMP-9 transcription start site, and suppresses MMP-9 transcription. Thus, HOXA2 acts as a suppressor or TBP-antagonist to inhibit MMP-9 expression; while methylation-mediated inactivation of HOXA2 in NPC derepresses MMP-9 production and increases invasion of NPC cells. In NPC plasma samples, increased plasma EBV copy number was correlated with increased in cell-free HOXA2 hypermethylation and elevated MMP-9 levels. Plasma EBV DNA and methylated cell-free HOXA2 can be used as biomarkers for monitoring NPC treatment.

Food allergy (FA) is an adverse immune response to certain innocent food. It is estimated about 2% to 6% of the general population suffer from FA. Symptoms of a food allergic reaction may involve the gastrointestinal tract or/and other organs. The gut microbiota plays a critical role in diet-induced health problems. Whether the changes in the composition of the intestinal microbiota regulate allergic responses to food remains poorly understood. Thus, we created an FA animal model, sequenced the V4-V5 regions of 16S rRNA genes to characterize the genera abundance of gut microbiota. The results showed that mice under FA condition showed different gut bacterial structures. Diverse distribution of the bacterial species was identified between FA and control groups. FA altered the components of intestinal Microbiota in mice. The dysbiosis of the gut metagenome correlated with the development of the FA.

Gallbladder cancer (GBC) is an aggressive malignancy. Although surgical resection may be curable, most patients are diagnosed at an advanced unresectable disease stage. Cholelithiasis is the major risk factor; however the pathogenesis of the disease, from gallstone cholecystitis to cancer, is still not understood. To understand the molecular genetic underpinnings of this cancer and explore novel therapeutic targets for GBC, we examined the key genes and pathways involved in GBC using RNA sequencing. We performed gene expression analysis of 32 cases of surgically-resected GBC along with normal gallbladder tissue controls. We observed that 519 genes were differentially expressed between GBC and normal GB mucosal controls. The liver X receptor (LXR)/retinoid X receptor (RXR) and farnesoid X receptor (FXR) /RXR pathways were the top canonical pathways involved in GBC. Key genes in these pathways, including SERPINB3 and KLK1, were overexpressed in GBC, especially in female GBC patients. Additionally, ApoA1 gene expression suppressed in GBC as compared with normal control tissues. LXR and FXR genes, known to be important in lipid metabolism also function as tumor suppressors and their down regulation appears to be critical for GBC pathogenesis. LXR agonists may have therapeutic value and as potential therapeutic targets.

MicroRNAs (miRNAs) play a pivotal role in carcinogenesis by suppressing oncogenes or tumor suppressor genes. Various studies have identified numerous miRNAs and their diverse targets; however, the consequences of dysregulated miRNAs in nasopharyngeal carcinoma (NPC) remain unclear. For this study, we found that miR-148a is downregulated through hypermethylation in NPC biopsies and NPC cell lines compared with adjacent normal and NP cells respectively. Promoter assays demonstrated that upstream stimulatory factor 1 (USF1) is a crucial transcription factor that activates miR-148a promoter activity. EMSA assays confirmed that purified USF1 binds better toward the unmethylated than the methylated CG-containing USF1 consensus probe. The ectopic expression of miR-148a inhibits cell migration in NPC cells through the suppression of integrin-mediated signaling by targeting VAV2, WASL and ROCK1. Biochemical and functional assays provided supporting evidence that these 3 genes are the downstream targets of miR-148a in NPC cells. Furthermore, immunohistochemical staining and Western blotting analysis revealed that the 3 oncogenic targets of miR-148a were overexpressed in NPC biopsies, suggesting that the inactivation of miR-148a caused by DNA methylation promotes NPC progression. Overall, our findings revealed that miR-148a can act as tumor suppressor miRNA and serve as a biomarker as well as a therapeutic target for NPC.