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Background: Multinucleate giant cells are described in breast aspirates. However, due to its rarity very few cases have been described cytologically. Hence recognition and correct interpretation of their presence is difficult, yet crucial for accurate diagnosis. Materials and Methods: The prospective study of FNAC (fine needle aspirate cytology) of breast lumps was conducted for a period of six months. Direct smears were prepared from the material aspirated. In case of fluid aspirates, centrifuge done and cell sediment was used for making smears. Smears were alcohol fixed and stained with PAP/H&E or air dried smears were stained with Leishman stain. Further smears were subjected to immunocytochemistry using vimentin and CD34 markers to know the origin of multinucleate giant cells. Results: We have reported 11 cases of breast lesions, which showed multinucleate giant cells on FNAC. Out of the 11 cases, Cytologically six cases showed granuloma debris with relative proportion of epithelioid histiocytes, lymphocytes, neutrophils and multinucleate giant cells. Two cases were diagnosed as acute suppurative granulomatous mastitis. Two cases of fibroadenoma and one case of fat necrosis showed multinucleate giant cells. Immunocytochemistry showed vimentin positivity in both stromal and histiocytic type of multinucleate giant cells and in isolated histiocytes. CD34 was focally positive in histiocytic type of giant cells. Conclusion: An effort is made to distinguish between the stromal and histiocytic type giant cells in non-neoplastic breast lesions. Further molecular studies have to be done to know the exact histogenesis and role of these multinucleate giant cells in benign lesions.

// Maria Raffaella Ambrosio 1,* , Lucia Mundo 1,* , Sara Gazaneo 1 , Matteo Picciolini 2 , Prasad Satya Vara 3 , Shaheen Sayed 4 , Alessandro Ginori 1,5 , Giuseppe Lo Bello 1 , Leonardo Del Porro 1 , Mohsen Navari 6,7,8 , Stefano Ascani 9 , Amhed Yonis 10 , Lorenzo Leoncini 1 , Pier Paolo Piccaluga 7,8,* and Stefano Lazzi 1,* 1 Department of Medical Biotechnology, Section of Pathology, University of Siena, Siena, Italy 2 Diatech Pharmacogenetics, Jesi, Italy 3 Aga Khan Hospital, Kisumu, Kenya 4 Aga Khan University Hospital, Nairobi, Kenya 5 Pathology Unit, Ospedale Civico di Carrara, Carrara, Italy 6 Research Center of Advanced Technologies in Medicine, Torbat Heydariyeh University of Medical Sciences, Torbat Heydariyeh, Iran 7 Department of Experimental, Diagnostic, and Experimental Medicine, Bologna University School of Medicine, Bologna, Italy 8 Euro-Mediterranean Institute of Science and Technology (IEMEST), Palermo, Italy 9 Section of Pathology, Azienda Ospedaliera S. Maria di Terni, University of Perugia, Perugia, Italy 10 Alexandria University, Alexandria, Egypt * These authors have contributed equally to this work Correspondence to: Lorenzo Leoncini, email: // Pier Paolo Piccaluga, email: // Keywords : plasmablastic lymphoma; miRNA expression profiling; Burkitt lymphoma; extramedullary plasmacytoma; Epstein-Barr virus; Pathology section Received : June 20, 2017 Accepted : September 08, 2017 Published : October 31, 2017 Abstract Plasmablastic lymphoma (PBL) is an aggressive lymphoma, often arising in the context of immunodeficiency and associated with Epstein-Barr virus (EBV) infection. The most frequently detected genetic alteration is the deregulation of MYC gene through the translocation - t(8;14)(q24;q32). The diagnosis of PBL is often challenging because it has an overlap in morphology, immunophenotype, cytogenetics and virus association with other lymphomas and plasma cell neoplasms; further, its molecular basis remains elusive. In the present study we aimed to better define the possible contribution of EBV infection as well as miRNA deregulation in PBL pathogenesis. We studied 23 cases of PBL, 19 Burkitt lymphomas (BL), and 17 extra-medullary plasmacytoma (EMPC). We used qPCR and immunohistochemistry to assess EBV latency patterns, while micro-RNA (miRNA) profiling was performed by next generation sequencing (Illumina) and validated by qPCR. Our analysis revealed a non-canonical EBV latency program with the partial expression of some proteins characterizing latency II and the activation of an abortive lytic cycle. Moreover, we identified miRNA signatures discriminating PBL from BL and EMPC. Interestingly, based on the miRNA profile, PBL appeared constituted by two discrete subgroups more similar to either BL or EMPC, respectively. This pattern was confirmed in an independent set of cases studied by qPCR and corresponded to different clinico-pathological features in the two groups, including HIV infection, MYC rearrangement and disease localization. In conclusion, we uncovered for the first time 1) an atypical EBV latency program in PBL; 2) a miRNA signature distinguishing PBL from the closest malignant counterparts; 3) the molecular basis of PBL heterogeneity.

// Dong Yeon Kim 1,* , Min-Kyung Kang 1,* , Sin-Hye Park 1 , Eun-Jung Lee 1 , Yun-Ho Kim 1 , Hyeongjoo Oh 1 , Yean-Jung Choi 1 and Young-Hee Kang 1 1 Department of Food and Nutrition, Hallym University, Chuncheon, Korea * These authors have contributed equally to this study Correspondence to: Young-Hee Kang, email: // Keywords : β-Catenin; cell-cell disjunction; eucalyptol; Snail; tubulointerstitial fibrosis; Pathology Section Received : June 16, 2017 Accepted : October 05, 2017 Published : October 16, 2017 Abstract Renal tubulointerstitial fibrosis is an important event in the pathogenesis of diabetic nephropathy. Under pathologic conditions, renal tubular epithelial cells undergo transition characterized by loss of cell-cell adhesion and increased cell migration. This study investigated that eucalyptol inhibited tubular epithelial cell disjunction and tubulointerstitial fibrosis stimulated by glucose. Human renal proximal tubular epithelial cells were incubated for up to 72 h in media containing 27.5 mM mannitol as osmotic controls or 33 mM glucose in the presence of 1-20 μM eucalyptol. Nontoxic eucalyptol inhibited glucose-induced expression of the mesenchymal markers of N-cadherin and α-smooth muscle actin, whereas the induction of E-cadherin was enhanced. Eucalyptol attenuated the induction of connective tissue growth factor and collagen IV by glucose, whereas the membrane type 1-matrix metalloproteinase expression was enhanced with reducing tissue inhibitor of metalloproteinase-2 expression. Oral administration of 10 mg/kg eucalyptol to db/db mice for 8 weeks blunted hyperglycemia and proteinuria. Eucalyptol reversed tissue levels of E-cadherin, N-cadherin and P-cadherin and the collagen fiber deposition in diabetic kidneys. Eucalyptol attenuated the induction of Snail1, β-catenin and integrin-linked kinase 1 (ILK1) in glucose-exposed tubular cells and diabetic kidneys, and the glycogen synthase kinase (GSK)-3β expression was reversely enhanced. Glucose prompted TGF-β1 production in tubular cells, leading to induction of Snail1, β-catenin and ILK1, which was dampened by eucalyptol. Furthermore, the Snail1 gene deletion encumbered the β-catenin induction in glucose/eucalyptol-treated tubular cells accompanying enhanced GSK-3β expression. Therefore, eucalyptol may antagonize hyperglycemia-induced tubular epithelial derangement and tubulointerstitial fibrosis through blocking ILK1-dependent transcriptional interaction of Snail1/β-catenin.

// Keisuke Nishiura 1,* , Naoki Ichikawa-Tomikawa 1,* , Kotaro Sugimoto 1 , Yasuto Kunii 2,3 , Korehito Kashiwagi 1 , Mizuko Tanaka 1 , Yuichi Yokoyama 4 , Mizuki Hino 2 , Takashi Sugino 5 , Hirooki Yabe 2 , Hitoshi Takahashi 4 , Akiyoshi Kakita 4 , Tetsuya Imura 1,6 and Hideki Chiba 1 1 Department of Basic Pathology, Fukushima Medical University School of Medicine, Fukushima, Japan 2 Department of Neuropsychiatry, Fukushima Medical University School of Medicine, Fukushima, Japan 3 Department of Psychiatry, Aizu Medical Center, Fukushima Medical University, Fukushima, Japan 4 Department of Pathology, Brain Research Institute, Niigata University, Niigata, Japan 5 Department of Diagnostic Pathology, Shizuoka Cancer Center, Shizuoka, Japan 6 Department of Surgical Pathology, Kyoto Prefectural University of Medicine, Kyoto, Japan * These authors contributed equally to this work Correspondence to: Hideki Chiba, email: // Keywords : blood-brain barrier; tight junction; claudin; protein kinase A; schizophrenia; Pathology Section Received : August 30, 2017 Accepted : October 04, 2017 Published : October 16, 2017 Abstract Schizophrenia is thought to be caused by a combination of genetic and environmental factors; however, its pathogenesis remains largely unknown. Here, we focus on the endothelial tight-junction protein claudin-5 (CLDN5), because the CLDN5 gene is mapped to the schizophrenia-associated 22q11.2 deletion region, and a single nucleotide polymorphism in the CLDN5 locus is also linked to schizophrenia. We show, by RT-qPCR and immunohistochemistry, that the expressions of CLDN5 mRNA and protein are significantly increased and decreased, respectively, in the schizophrenic prefrontal cortex (PFC) compared with control PFC. These changes were not observed in the schizophrenic visual cortex (VC), and neither the density nor diameter of the CD34-positive microvessels was altered in the schizophrenic PFC or VC. Interestingly, protein kinase A (PKA) was activated in the microvascular and perivascular regions of the schizophrenic PFC, and the pPKA-positive microvascular endothelial cells occasionally exhibited focal loss of CLND5. Since we previously demonstrated that cAMP induced CLDN5 mRNA expression and size-selective loosening of the endothelial barrier in PKA-independent and -dependent manners, respectively, a similar mechanism could contribute to the discrepancy between mRNA and protein expression of CLDN5 in the schizophrenic PFC. Taken collectively, these findings provide novel insights into the pathophysiology of schizophrenia.

// Hiroshi Yoshida 1,* , Yoshiko Yamashita 2,* , Taichi Shimazu 3 , Eric Cosatto 4 , Tomoharu Kiyuna 2 , Hirokazu Taniguchi 1 , Shigeki Sekine 1,5 and Atsushi Ochiai 1,5,6 1 Division of Pathology and Clinical Laboratories, National Cancer Center Hospital, Chuo-ku, Tokyo, Japan 2 Medical Solutions Division, NEC Corporation, Minato-ku, Tokyo, Japan 3 Epidemiology and Prevention Group, Research Center for Cancer Prevention and Screening, National Cancer Center, Chuo-ku, Tokyo, Japan 4 Department of Machine Learning, NEC Laboratories America, Princeton, NJ, USA 5 Division of Molecular Pathology, National Cancer Center Research Institute, Chuo-ku, Tokyo, Japan 6 Division of Pathology, Research Center for Innovative Oncology, National Cancer Center, Kashiwa, Chiba, Japan * These authors are equally contributed to this work Correspondence to: Hiroshi Yoshida, email: // Keywords : colorectal cancer, artificial intelligence, automated image analysis, whole slide imaging, histological classification, Pathology Section Received : March 20, 2017 Accepted : September 30, 2017 Published : October 12, 2017 Abstract Background: An automated image analysis system, e-Pathologist, was developed to improve the quality of colorectal biopsy diagnostics in routine pathology practice. Objective: The aim of the study was to evaluate the classification accuracy of the e-Pathologist image analysis software in the setting of routine pathology practice in two institutions. Materials and methods: In total, 1328 colorectal tissue specimens were consecutively obtained from two hospitals (1077 tissues from Tokyo hospital, and 251 tissues from East hospital) and the stained specimen slides were anonymized and digitized. At least two experienced gastrointestinal pathologists evaluated each slide for pathological diagnosis. We compared the 3-tier classification results (carcinoma or suspicion of carcinoma, adenoma, and lastly negative for a neoplastic lesion) between the human pathologists and that of e-Pathologist. Results: For the Tokyo hospital specimens, all carcinoma tissues were correctly classified (n=112), and 9.9% (80/810) of the adenoma tissues were incorrectly classified as negative. For the East hospital specimens, 0 out of the 51 adenoma tissues were incorrectly classified as negative while 9.3% (11/118) of the carcinoma tissues were incorrectly classified as either adenoma, or negative. For the Tokyo and East hospital datasets, the undetected rate of carcinoma, undetected rate of adenoma, and over-detected proportion were 0% and 9.3%, 9.9% and 0%, and 36.1% and 27.1%, respectively. Conclusions: This image analysis system requires some improvements; however, it has the potential to assist pathologists in quality improvement of routine pathological practice in the not too distant future.

// Hongwei Cheng 1,* , Xiang Mao 1,2,5,* , Zonggang Hou 3,5,* , Jian Xu 2 , Shuyu Hao 3,5 , Huan Li 3,5 and Baiyun Liu 2,3,4,5 1 Department of Neurosurgery, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, China 2 Neurotrauma Laboratory, Beijing Neurosurgical Institute, Capital Medical University, Beijing, China 3 Department of Neurosurgery, Beijing Tian Tan Hospital, Capital Medical University, Beijing, China 4 Nerve Injury and Repair Center of Beijing Institute for Brain Disorders, Beijing, China 5 China National Clinical Research Center for Neurological Diseases, Beijing, China * These authors contributed equally to this work Correspondence to: Baiyun Liu, email: // Keywords : cerebral perfusion pressure, regional cerebral blood flow, intracranial pressure, contrast-enhanced ultrasound, Pathology Section Received : July 31, 2017 Accepted : September 22, 2017 Published : October 11, 2017 Abstract To discuss the relationship between the regional cerebral blood flow (rCBF) and cerebral perfusion pressure (CPP) and the effect of CPP on rCBF in different spaces in an experimental animal model. As the ICP increased, the CPP and rCBF (A × β value measured by CEU) decreased to varying degrees. The rCBF 1 and rCBF 2 were well correlated with the CPP. At the same CPP, rCBF 1 decreased significantly than the level of rCBF 2 (p < 0.01). Six healthy cross-breed dogs, both males and females, weighing 18.3 ± 1.6 kg, were selected to establish models of increased intracranial pressure (ICP) via the installation of an epidural latex sacculus. The calculated CPP was in accordance with the ICP through the formula CPP = MAP - ICP, and contrast-enhanced ultrasound (CEU) was used to instantly measure the rCBF 1 and 2 cm around the sacculus edge. The relationship between rCBF 1 cm (rCBF 1 ) and 2 cm (rCBF 2 ) around the sacculus edge and the CPP was analyzed. As the ICP increased, the CPP and rCBF both decreased. The rCBF and the CPP had a linear relationship, but the perfusion pressure did not necessarily determine all parts of the rCBF. The rCBF was different in different spaces: the farther away from the injured site, the smaller the effect on the rCBF.

// Maria Ramnefjell 1 , Christina Aamelfot 2 , Lars Helgeland 3 and Lars A. Akslen 1,3 1 Centre for Cancer Biomarkers CCBIO, Department of Clinical Medicine, Section for Pathology, Haukeland University Hospital, University of Bergen, Bergen, Norway 2 Department of Thoracic Medicine, Haukeland University Hospital, Bergen, Norway 3 Department of Pathology, Haukeland University Hospital, Bergen, Norway Correspondence to: Lars A. Akslen, email: // Keywords : lung cancer, metastases, SerpinB2, neuroserpin, L1CAM, Pathology Section Received : March 16, 2017 Accepted : September 22, 2017 Published : October 03, 2017 Abstract Lung cancer is a leading cause of cancer deaths worldwide and new biomarkers are of utmost importance. Studies have indicated that the anti-plasminogen activators SerpinB2 and Neuroserpin, and the adhesion molecule L1CAM, have a coordinated impact on development of metastasis. Here, we examined whether expression of these markers was associated with clinico-pathologic characteristics and prognosis in resected non-small cell lung cancer (NSCLC). Surgical specimens from 438 NSCLC patients treated at Haukeland University Hospital, Bergen, Norway (1993-2010) were included (median age 68 years; 213 adenocarcinomas, 135 squamous cell carcinomas, 90 others). Representative tumor sections were stained for SerpinB2, Neuroserpin, and L1CAM. Low expression of SerpinB2 was associated with reduced lung cancer specific survival (LCSS) in adenocarcinomas ( p = 0.017), also in stage I ( p = 0.031). In contrast, high SerpinB2 was associated with reduced LCSS in stage I squamous cell carcinomas ( p = 0.022). Although Neuroserpin and L1CAM showed some associations with clinico-pathologic phenotype, there were no associations with survival. In multivariate survival analysis of adenocarcinomas, low SerpinB2 demonstrated independent prognostic value (HR 1.8, p = 0.008). In summary, low expression of SerpinB2 in lung adenocarcinomas was an independent prognostic factor. In contrast to findings by others, we found no impact of L1CAM on survival.

// Onju Ham 1 , Se-Yeon Lee 1 , Byeong-Wook Song 2 , Chang Youn Lee 3 , Jiyun Lee 1 , Hyang-Hee Seo 1 , Sang Woo Kim 4 , Soyeon Lim 3 , Il-Kwon Kim 4 , Seahyoung Lee 4 and Ki-Chul Hwang 4 1 Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul, Republic of Korea 2 EIT/LOFUS R&D Center, International St. Mary’s Hospital, Incheon, Republic of Korea 3 Department of Integrated Omics for Biomedical Sciences, Graduate School, Yonsei University, Seoul, Republic of Korea 4 Institute for Bio-Medical Convergence, College of Medicine, Catholic Kwandong University, Incheon, Republic of Korea Correspondence to: Seahyoung Lee, email: // Ki-Chul Hwang, email: // Keywords : miR-9, PDGFR, small molecule, neointima, Pathology Section Received : March 08, 2017 Accepted : September 04, 2017 Published : September 28, 2017 Abstract Pathologic proliferation and migration of vascular smooth muscle cells (VSMCs) exacerbate cardiovascular disease. MicroRNAs (miRNAs), as endogenous inhibitors of protein synthesis, are expected to modulate pathologic proliferation of VSMCs. Here we report that both platelet-derived growth factor receptor (PDGFR) targeting miR-9 and a small molecule that increases miR-9 can inhibit the serum-induced proliferation of VSMCs. First, based on miRNA-target prediction databases and empirical data, we have selected miR-9 as a potent anti-proliferative miRNA in VSMCs. Further examination indicated that miR-9 directly targets PDGFR disrupting downstream signaling cascades, and this resulted in inhibition of VSMC proliferation and migration. Exogenous delivery of miR-9 inhibited VSMC proliferation in vitro, and a small molecule that increased miR-9 expression also inhibited neointima formation following balloon injury in vivo . We provide evidence of miRNA-mediated modulation of VSMC proliferation and further demonstrate that small molecule-mediated regulation of miRNA targeting a key regulator of VSMC proliferation is a viable therapeutic strategy for treating vascular disease involving pathologic VSMC proliferation such as restenosis.

// Dazhi Deng 1,2 , Chongdong Jian 2,3 , Ling Lei 2,4 , Yijing Zhou 2 , Colleen McSweeney 2 , Fengping Dong 2 , Yilun Shen 2 , Donghua Zou 5 , Yonggang Wang 6 , Yuan Wu 3 , Limin Zhang 7 and Yingwei Mao 2 1 Department of Emergency, The People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, Guangxi, China 2 Department of Biology, Pennsylvania State University, University Park, PA, USA 3 Department of Neurology, First Affiliated Hospital, Guangxi Medical University, Nanning, Guangxi, China 4 Health Examination Center, The People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, Guangxi, China 5 Department of Neurology, The First People’s Hospital of Nanning, Nanning, Guangxi, China 6 Department of Neurology, School of Medicine, Renji Hospital, Shanghai Jiaotong University, Shanghai, China 7 CAS Key Laboratory of Magnetic Resonance in Biological Systems, State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, National Centre for Magnetic Resonance in Wuhan, Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences, Wuhan, China Correspondence to: Limin Zhang, email: // Yingwei Mao, email: // Keywords : DISC1, interneuron, neural progenitors, metabolism, depression, Pathology Section Received : March 14, 2017 Accepted : September 03, 2017 Published : September 28, 2017 Abstract Mental illnesses like schizophrenia (SCZ) and major depression disorder (MDD) are devastating brain disorders. The SCZ risk gene, disrupted in schizophrenia 1 ( DISC1 ), has been associated with neuropsychiatric conditions. However, little is known regarding the long-lasting impacts on brain metabolism and behavioral outcomes from genetic insults on fetal NPCs during early life. We have established a new mouse model that specifically interrupts DISC1 functions in NPCs in vivo by a dominant-negative DISC1 (DN-DISC1) with a precise temporal andspatialregulation. Interestingly, prenatal interruption of mouse Disc1 function in NPCs leads to abnormal depression-like deficit in adult mice. Here we took a novel unbiased metabonomics approach to identify brain-specific metabolites that are significantly changed in DN-DISC1 mice. Surprisingly, the inhibitory neurotransmitter, GABA, is augmented. Consistently, parvalbumin (PV) interneurons are increased in the cingulate cortex, retrosplenial granular cortex, and motor cortex. Interestingly, somatostatin (SST) positive and neuropeptide Y (NPY) interneurons are decreased in some brain regions, suggesting that DN-DISC1 expression affects the localization of interneuron subtypes. To further explore the cellular mechanisms that cause this change, DN-DISC1 suppresses proliferation and promotes the cell cycle exit of progenitors in the medial ganglionic eminence (MGE), whereas it stimulates ectopic proliferation of neighboring cells through cell non-autonomous effect. Mechanistically, it modulates GSK3 activity and interrupts Dlx2 activity in the Wnt activation. In sum, our results provide evidence that specific genetic insults on NSCs at a short period of time could lead to prolonged changes of brain metabolism and development, eventually behavioral defects.

// Yoon Jin Cha 1 and Hyo Sup Shim 1 1 Department of Pathology, Severance Hospital, Yonsei University College of Medicine, Seoul, Korea Correspondence to: Hyo Sup Shim, email: shimhs@yuhs.ac Keywords : myofibroblastic tumor, inflammatory, anaplastic lymphoma kinase, PD-L1, lymphocyte, Pathology Section Received : April 08, 2017 Accepted : July 12, 2017 Published : September 15, 2017 Abstract Background : Inflammatory myofibroblastic tumors (IMTs) are rare mesenchymal neoplasms that are composed of myofibroblastic cells accompanied by inflammatory infiltrate. We investigated the immune profiles of IMTs, including PD-L1 expression and proportion of CD8+ tumor-infiltrating lymphocytes (TILs), as well as its clinicopathological characteristics according to ALK gene rearrangementstatus. Methods : Twenty-eight IMTs from 25 patients were retrieved from our pathology files (2005–2015), and their clinicopathological parameters and outcomes were analyzed. Immunohistochemistry (IHC) was performed using whole-tissue sections to detect PD-L1 and CD8 expression, and fluorescent in situ hybridization (FISH) analysis and IHC were performed using tissue microarrays to identify rearrangements in the ALK, ROS1 , and RET genes. Results : ALK rearrangement was observed in 11 cases (44.0%), and all cases exhibited diffuse cytoplasmic ALK expression during IHC. ROS1 or RET rearrangement was not detected using IHC or FISH. IMTs harboring ALK rearrangement ( ALK -positive) were located in the lungs ( n = 7), genitourinary tract ( n = 2), and mesentery ( n = 1). The mean patient age was 33.2 years for ALK -positive IMTs and 53.1 years for ALK -negative IMTs. All patients with ALK -positive IMTs survived without recurrence or metastasis. IMTs with metastasis and/or recurrence were ALK -negative and exhibited elevated PD-L1 expression (positive tumor cells: 70.0% vs . 21.3%, P = 0.023; H-score: 107.5 vs . 26.3, P = 0.005). In addition, ALK -negative IMTs had a more CD8+ TILs, compared to ALK -positive IMTs (23.3% vs . 8.9%, P = 0.027). Conclusion : ALK -positive IMTs are characterized by younger age, well-defined margins, frequent involvement of the lung, and fewer CD8+ TILs. Greater PD-L1 expression was observed in IMTs with tumor necrosis and metastasis/recurrence, which were also negative for ALK rearrangement. These results suggest that immune checkpoint inhibitors may be a novel option for treating patients with advanced IMT.

// Xu Tian 1,* , Xiaohui Xu 2,* , Kai Zhang 3 and Hui Wang 2 1 College of Economics and Management, China Center for Food Security Studies, Nanjing Agricultural University, Nanjing, Jiangsu, China 2 Department of Epidemiology and Biostatistics, School of Public Health, Nanjing Medical University, Nanjing, Jiangsu, China 3 Pancreatic Center and Department of General Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing,Nanjing, Jiangsu, China * These authors have contribute equally to this study Correspondence to: Hui Wang, email: // Keywords : age, sex, dietary diversity score, metabolic syndrome, Pathology Section Received : July 29, 2017 Accepted : August 24, 2017 Published : September 02, 2017 Abstract Background: Previous research indicated that dietary diversity had favorable association with metabolic syndrome (MetS), and it has not been investigated in China. Methods: Adults (aged 18+) with complete dietary and biochemical data were collected from 2009 China Health and Nutrition Survey ( n =4308). Dietary diversity was measured by modified Dietary Diversity Score (DDS). MetS was defined by the harmonized criteria. The association between DDS and MetS was investigated by multivariable adjusted logistic regression. Results: An inverse-U shape relationship between MetS risk and age was detected for both genders, and female were more vulnerable than male at old times. More diversified diet decreased the risk of MetS for young female (≥18 & ≤45), similar trends were detected in serum TGs, abdominal adiposity, blood pressure, and fasting blood glucose (all P 60) and male adults (>45&≤60). Greater DDS was associated with higher serum TGs, and lower HDL-C level for male adults, higher blood pressure for old men, but lower blood pressure and fasting blood glucose in young men (all P

// Joong Sun Kim 1 , Na-Kyung Han 2 , Sung-Ho Kim 3 and Hae-June Lee 2 1 Research Center, Dongnam Institute of Radiological & Medical Sciences, Busan, Korea 2 Division of Basic Radiation Bioscience, Korea Institute of Radiological and Medical Sciences, Seoul, Korea 3 College of Veterinary Medicine, Chonnam National University, Gwangju, Korea Correspondence to: Hae-June Lee, email: // Keywords : silibinin, fibrosis, EMT, TGF-β1, radiation, Pathology Section Received : July 12, 2017 Accepted : August 26, 2017 Published : September 02, 2017 Abstract Radiotherapy is a common treatment for cancer patients, but its use is often restricted by the tolerance of normal tissue. As cancer patients live longer, delayed radiation effects on normal tissue have become a concern. Radiation-induced enteropathy, including inflammatory bowel disease and fibrosis, are major issues for long-term cancer survivors. To investigate whether silibinin attenuates delayed radiation-induced intestinal injury in mice, we focused on intestinal fibrotic changes. Silibinin improved delayed radiation injuries in mice in association with decreased collagen deposition within the intestines and deceased transforming growth factor (TGF)-β1 levels in the intestine and plasma. Treating mice bearing CT26 mouse colon cancer tumors with both silibinin and radiation stimulated tumor regression more than radiation alone. We also investigated the effect of silibinin on the radiation-induced epithelial-to-mesenchymal transition (EMT), the primary mechanism of fibrosis. We assessed changes in E-cadherin, N-cadherin, and α-smooth muscle actin expression, and demonstrated that silibinin attenuates radiation-induced EMT. Irradiating intestinal epithelial cells increased TGF-β1 levels, but silibinin suppressed TGF-β1 expression by inhibiting Smad2/3 phosphorylation. These results suggest silibinin has the potential to serve as a useful therapeutic agent in patients with radiation-induced intestinal fibrosis.

// Lingling Chen 1,2,* , Chunyan Liu 1,* , Jianfeng Gao 1 , Zhiwen Xie 3 , Lawrence W.C. Chan 4 , Damien J. Keating 5 , Yibin Yang 6 , Jiazhong Sun 7 , Fuling Zhou 8 , Yongchang Wei 9 , Xiuli Men 10 and Sijun Yang 1 1 ABSL-3 Laboratory at the Center for Animal Experiment and Institute of Animal Model for Human Disease, Wuhan University School of Medicine, Wuhan, P. R. China 2 Department of Cell Biology, College of Life Science, Nanjing Normal University, Nanjing, Jiangsu, P.R. China 3 School of Bioscience and Technology , Weifang Medical University, Weifang Shandong, P.R. China 4 Department of Health Technology and Informatics, Hong Kong Polytechnic University, Hong Kong, Hong Kong 5 Department of Human Physiology and Centre for Neuroscience, Flinders University, Adelaide, South Australia, Australia 6 Department of Endocrinology, Zhongnan Hospital of Wuhan University, Wuhan, Hubei, P.R. China 7 Department of Respiratory, Zhongnan Hospital of Wuhan University, Wuhan, Hubei, P.R. China 8 Department of Hematology and Radiation, Zhongnan Hospital of Wuhan University, Wuhan, Hubei, P.R. China 9 Department of Medical Oncology, Zhongnan Hospital of Wuhan University, Wuhan, Hubei, P.R. China 10 School of Basic Medical Sciences, North China University of Science and Technology, Tangshan, P.R. China * These authors have contributed equally to this work Correspondence to: Sijun Yang, email: // Keywords : Miro1, mitophagy, insulin resistance, HFD, diabetes, Pathology Section Received : June 26, 2017 Accepted : August 29, 2017 Published : September 16, 2017 Abstract Mitochondrial function is essential to meet metabolic demand of pancreatic beta cells respond to high nutrient stress. Mitophagy is an essential component to normal pancreatic β-cell function and has been associated with β-cell failure in Type 2 diabetes (T2D). Our previous studies have indicated that mitochondrial Rho (Miro) GTPase-mediated mitochondrial dysfunction under high nutrient stress leads to NOD-like receptor 3 (NLRP3)-dependent proinflammatory responses and subsequent insulin resistance. However, the in vivo mechanism by which Miro1 underlies mitophagy has not been identified. Here we show firstly that the expression of Miro is reduced in human T2D and mouse db/db islets and in INS-1 cell line exposed to high glucose and palmitate. β-cell specific ablation of Miro1 (Miro1f/f: Rip-cre mice, or (IKO) under high nutrient stress promotes the development of hyperglycemia. β-cells from IKO mice display an inhibition of mitophagy under oxidative stress and induces mitochondrial dysfunction. Dysfunctional mitophagy in IKO mice is represented by damaged islet beta cell mitochondrial and secretory capacity, unbalanced downstream MKK-JNK signalling without affecting the levels of MEK, ERK or p38 activation and subsequently, impaired insulin secretion signaling via inhibition IRS-AKT-Foxo1 pathway, leading to worsening glucose tolerance in these mice. Thus, these data suggest that Miro1 may be responsible for mitophagy deficiency and β-cell dysfunction in T2D and that strategies target Miro1 in vivo may provide a therapeutic target to enhance β-cell mitochondrial quality and insulin secretion to ameliorate complications associated with T2D.

// Jianxin Diao 1,* , Haiye Li 1,* , Wei Huang 1,* , Wenxiao Ma 2 , Huan Dai 1 , Yawei Liu 3 , Ming Wang 4 , He Yu Hua 1 , Jinying Ou 1 , Xiaomin Sun 1 , Xuegang Sun 1 and Yungao Yang 1 1 School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, China 2 Gao Ming People’s Hospital, Foshan, Guangdong, China 3 Nanfang Hospital, Southern Medical University, Guangdong,Guangzhou, China 4 Zhujiang Hospital of Southern Medical University, Guangdong, Guangzhou, China * These authors have contributed equally to this work Correspondence to: Yungao Yang, email: // Xuegang Sun, email: // Xiaomin Sun, email: // Keywords : San huang yin chi decoction, acute on chronic liver failure, APE1/Ref-1, apoptosis, p53, Pathology Section Received : November 15, 2016 Accepted : January 24, 2017 Published :August 04, 2017 Abstract Background & Aims: San huang yin chi decoction(SHYCD) is derived from the yin chen hao decoction, a well-known and canonical Chinese medicine formula from the “Treatise on Febrile Diseases”. Over the past decade, SHYCD has been used to treat and prevent the liver cirrhosis and liver failure. In the present study, we investigated the effects of SHYCD for acute on chronic liver failure(ACLF) and explored its potential mechanism. an ACLF rat model, which induced by carbon tetrachloride (CCl4) combined with D-galactosamine (D-GalN) and lipopolysaccharide(LPS), was used and confirmed by B-ultrasound analysis. Rats were randomly divided into control group, model group, SHYCD-H group, SHYCD-M group, SHYCD-L group, AGNHW group. Compared with the ACLF model group, High, medium, and low doses of SHYCD reduced ALT, AST, TBIL, NH3, IL-1β, IL-6, and TNFα expression levels in the serum, Shorten PT and INR time,and increased Fbg content in the whole blood, increased survival rate of the rats, improved liver pathological changes. APE1 / Ref-1 was mainly expressed in the nucleus, but the nucleus and cytoplasm were co-expressed after hepatocyte injury. SHYCD significantly downregulated APE1/Ref-1 expression in the cytoplasm. Increased APE1/Ref-1, Bcl-2, reduced p53, caspase-3, Bax, and Cyt-c in the total protein. Base on the results, we conclused that High, medium, and low doses of SHYCD could be applied in prevention and treatment of ACLF, and dose-dependent. The possible mechanism is to promote the APE1 / Ref-1 from the cytoplasm to the nuclear transfer, regulation of p53 apoptosis signal pathway prevention and treatment of ACLF.

This study presents a rational design approach to discovery synthetic peptide vaccine candidates from endogenous proteins for chronic non-infectious diseases immunological therapeutics. The approach described the screening of key antigenic amino acid residues of the interleukine-13, which is up-regulated expression in asthma, followed by the development of immunological helper epitope peptides via an integrative computational and experimental method. Notably, this totally synthetic peptide vaccine was capable of stimulating humoral immune responses much stronger than those of parental antigenic peptides by enhancing the efficiency of antigen presentation, and had effective treatment in mouse asthma models. Our approach offers new possibilities to discovery therapeutic peptide vaccine candidates for chronic non-infectious diseases, with highly consolidated in silico and animal disease models for fast iterative screening.

// Kuan Li 1,* , Qi Wu 2,* , Xin Sun 2,* , Yan Geng 3 , Dong Leng 4 , Hongwei Li 5 , Subei Zhang 2 , Qiaoxing Wang 2 , Junping Wu 5 , Long Xu 5 , Xue Li 1 , Yu Li 1 , Qiuyang Zhang 1 , Adrianne Kurkciyan 3 , Jiurong Liang 3 , Dianhua Jiang 3 and Huaiyong Chen 1,2 1 Department of Basic Medicine, Haihe Clinic College of Tianjin Medical University, Tianjin, China 2 Key Research Laboratory for Infectious Disease Prevention for State Administration of Traditional Chinese Medicine, Tianjin Institute of Respiratory Diseases, Tianjin Haihe Hospital, Tianjin, China 3 Department of Medicine, Division of Pulmonary and Critical Care Medicine, Women’s Guild Lung Institute, Cedars-Sinai Medical Center, Los Angeles, California, United States 4 Clinical Laboratory, Beijing Chao-Yang Hospital, Capital Medical University, Beijing, China 5 Department of Respiratory Medicine, Tianjin Haihe Hospital, Tianjin, China * These authors have contributed equally to this work Correspondence to: Huaiyong Chen, email: // Dianhua Jiang, email: // Keywords : AT2 cells, intralobar pulmonary sequestration, endothelial, Tsp1, CD36, Pathology Section Received : February 15, 2017 Accepted : July 20, 2017 Published : August 04, 2017 Abstract An aberrant systemic artery supply results in recurrent infections in the abnormal lung lobe of intralobar pulmonary sequestration (ILS). The mechanisms underlying such persistent inflammation are unknown. Here, we hypothesize that alteration of an endothelial cell niche for alveolar epithelial cells results in the impaired proliferation potential of alveolar progenitor cells, leading to the defective defense mechanism in intralobar pulmonary sequestration. Paraffin sections of lung tissues from patients with intralobar pulmonary sequestration or from healthy controls were collected for analysis of alveolar epithelial alterations in intralobar pulmonary sequestration by quantitative RT-PCR or immunofluorescent staining. Differential transcripts were identified between human pulmonary artery endothelial cells and human aortic endothelial cells by microarray. Validation of microarray data by quantitative PCR analysis indicated that thrombospondin-1 expression level is low in near-lesion part but high in lesion part of ILS lobe as compared to healthy controls. In vitro 3-D matrigel culture was adopted to evaluate the regulation of alveolar progenitor cells by thrombospondin-1 and CD36. We found that the proliferative potential of alveolar type 2 stem/progenitor cells was impaired in intralobar pulmonary sequestration. Mechanistically, we discovered that endothelial thrombospondin-1 promotes alveolar type 2 cell proliferation through the interaction with CD36. These data demonstrate that alveolar stem cells are impaired in the abnormal lobe from patients with intralobar pulmonary sequestration and imply that restoring epithelial integrity can be beneficial for the future treatments of recurrent infections in lung pathologies.

// Ligen Shi 1,* , Jianwei Lei 1,* , Hangzhe Xu 1 , Jingwei Zheng 1 , Yan Wang 1 , Yucong Peng 1 , Jun Yu 1 and Jianmin Zhang 1,2,3 1 Department of Neurosurgery, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China 2 Brain Research Institute, Zhejiang University, Hangzhou, Zhejiang, China 3 Collaborative Innovation Center for Brain Science, Zhejiang University, Hangzhou, Zhejiang, China * These authors have contributed equally to this work Correspondence to: Jun Yu, email: // Jianmin Zhang, email: // Keywords : hydrogen sulfide, MST1, neuronal apoptosis, subarachnoid hemorrhage, early brain injury, Pathology Section Received : July 03, 2017 Accepted : August 08, 2017 Published : August 28, 2017 Abstract Background: Hydrogen sulfide (H 2 S) has shown a neuroprotective role in several cerebrovascular diseases. This study aimed to explore the underlying mechanisms of H 2 S in early brain injury after subarachnoid hemorrhage (SAH). Methods: One hundred seventy-seven male Sprague-Dawley rats were employed in this study. Sodium hydrosulfide (NaHS), a donor of H 2 S, was injected intraperitoneally at 60 min after SAH was induced by endovascular perforation. Western blot analysis determined the expression of several proteins of interest, and an immunofluorescence assay was used to examine neuronal apoptosis. Results: Exogenous NaHS markedly improved neurological scores, attenuated brain edema, and ameliorated neuronal apoptosis at 24 h after SAH induction. The underlying mechanisms of H 2 S in ameliorating neuronal apoptosis might be executed through inhibition of the activity of mammalian sterile 20-like kinase 1 (MST1) protein. Western blot analysis demonstrated that exogenous NaHS decreased cleaved MST1 (cl-MST1) while increasing full-length MST1 expression. This anti-apoptotic effect of H 2 S could be reversed by chelerythrine, which could activate MST1 via caspase-dependent cleavage. Conclusions: Exogenous NaHS, as a donor of H 2 S, could ameliorate early brain injury after SAH by inhibiting neuronal apoptosis by reducing the activity of the MST1 protein.

// Keke Wang 1,* , Xiaojiang Deng 2,* , Zhihua Shen 1 , Yanan Jia 1 , Ranran Ding 3 , Rujia Li 1 , Xiaomin Liao 1 , Sisi Wang 1 , Yanping Ha 1 , Yueqiong Kong 4 , Yuyou Wu 4 , Junli Guo 4 and Wei Jie 1 1 Department of Pathology, School of Basic medicine Sciences, Guangdong Medical University, Zhanjiang, P.R. China 2 Department of Cardiovascular, Nanfang Hospital, Southern Medical University, Guangzhou, P.R. China 3 Department of Pathology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, P.R. China 4 Cardiovascular Institute of 1st Affiliated Hospital & Key Laboratory of Tropical Diseases and Translational Medicine of Ministry of Education, Hainan Medical University, Haikou, P.R. China * Joint first authors Correspondence to: Junli Guo, email: // Wei Jie, email: // Keywords : high glucose, vascular smooth muscle cell, Pim-1, cell proliferation, STAT3 signaling, Pathology Section Received : March 09, 2017 Accepted : June 28, 2017 Published : July 18, 2017 Abstract Serine/threonine kinase proviral integration site for Moloney murine leukemia virus 1 (Pim-1) plays an essential role in arterial wall cell proliferation and associated vascular diseases, including pulmonary arterial hypertension and aortic wall neointima formation. Here we tested a role of Pim-1 in high-glucose (HG)-mediated vascular smooth muscle cell (VSMC) proliferation. Pim-1 and proliferating cell nuclear antigen (PCNA) expression levels in arterial samples from streptozotocin-induced hyperglycemia rats were increased, compared with their weak expression in normoglycemic groups. In cultured rat VSMCs, HG led to transient Pim-1 expression decline, followed by sustained expression increase at both transcriptional and translational levels. Immunoblot analysis demonstrated that HG increased the expression of the 33-kDa isoform of Pim-1, but at much less extent to its 44-kDa plasma membrane isoform. D-glucose at a concentration of 25 mmol/L showed highest activity in stimulating Pim-1 expression. Both Pim-1 inhibitor quercetagetin and STAT3 inhibitor stattic significantly attenuated HG-induced VSMC proliferation and arrested cell cycle progression at the G1 phase. Quercetagetin showed no effect on Pim-1 expression but decreased the phosphorylated-Bad (T112)/Bad ratio in HG-treated VSMCs. However, stattic decreased phosphorylated-STAT3 (Y705) levels and caused transcriptional and translational down-regulation of Pim-1 in HG-treated VSMCs. Our findings suggest HG-mediated Pim-1 expression contributes to VSMC proliferation, which may be partly due to the activation of STAT3/Pim-1 signaling.

Autogenous bone graft is the best for spinal fusion in clinics, however, lacking sources, bleeding and infection are limited its practice. Seeking alternative materials are urgent for orthopaedic surgeon. Here, we evaluated osteoblast-oriented differentiation of rabbit BMSCs by co-culturing with composite scaffolds constructed using silicon-substituted-CaP-fine particulate bone powder-alginate. Using CCk8-kit, biocompatibility was evaluated by testing BMSCs proliferation; morphology and survival of osteoblasts within scaffolds were observed using EM and HE staining; growth factors and related genes were detected using RT-PCR. HE staining showed spindle-shaped BMSCs after the 3rd passage; EM data showed that uneven surface and longitudinal section were observed with scattered distribution of 5-100 mm interspaces, which leave enough space for BMSCs adhesion and growth. Interestingly, at 14-day culture with HE staining, osteocytes within the scaffolds grew well with regular shape and integrate structure. RT-PCR results showed that expression levels of BMP2, TGF-b and COL-I, ALP, OPN were increased significantly and time-dependently. Collectively, all mentioned effects were more obvious in co-culture BMSCs with scaffolds than those with other components. Immunohistochemistry showed that positive OPN expression was detected at 7-day co-culturing BMSCs with scaffold, rather than other situations. These results suggest that composite scaffolds constructed with Si-CaP-fine particulate bone powder-alginate have a certain degree of biocompatibility and bioactivity to promote osteoblast-oriented BMSCs differentiation.

// Xumin Ou 1,2,* , Sai Mao 1,2,* , Jingyu Cao 1,2 , Yunchao Ma 1,2 , Guangpeng Ma 4 , Anchun Cheng 1,2,3,* , Mingshu Wang 1,2,3,* , Dekang Zhu 2,3 , Shun Chen 1,2,3 , Renyong Jia 1,2,3 , Mafeng Liu 1,2,3 , Kunfeng Sun 1,2,3 , Qiao Yang 1,2,3 , Ying Wu 1,2,3 and Xiaoyue Chen 2,3 1 Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu, Sichuan, People′s Republic of China 2 Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Wenjiang, Chengdu, Sichuan, People′s Republic of China 3 Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu, Sichuan, People′s Republic of China 4 China Rural Technology Development Center, Beijing, People′s Republic of China * These authors have contributed equally to this work Correspondence to: Anchun Cheng, email: // Mingshu Wang, email: // Keywords : duck hepatitis A virus, viral hepatitis, liver injury, pathogenesis, viral-host interaction, Pathology Section Received : February 17, 2017 Accepted : June 20, 2017 Published : July 05, 2017 Abstract Duck Hepatitis A Virus (DHAV) belongs to the Avihepatovirus , which is also classified into Picornaviridae with Hepatovirus , Hepatitis A Virus (HAV). In humans, the pathogenesis of HAV is not well understood because of limited work with animal models. Here, we investigated the progress of duck viral hepatitis caused by DHAV and their potential for dissecting the pathogenesis of HAV. During the course of infection, the duck model had undergone hepatocellular lesions (vacuolation, acidophilic degeneration and steatosis), lymphocytes recruitment (neutrophil granulocytes, heterophilic granulocytes and T cells or plasm cells) and repair (activation of hepatic stellate cells, fibrosis and regeneration). Coincident with liver injury, the serum biomarkers, aspartate aminotransferase and alanine transaminase were significantly increased. Moreover, comparatively lower CD4+ and CD8+ T-cells were recruited to the liver, which might lead to a persistent infection (40 wk). Because DHAV and HAV have similar genomic structure, biological phenotypes and can easily replicate in liver. And half of fibrosis-related genes had high homology between humans and ducks. Considering these similarity in pathological and virological phenotypes, we proposed that the ducks might be an alternatively small animal model that would provide insight into the pathogenesis of viral hepatitis, fibrosis and liver regeneration.

// Meisheng Zou 1,2,* , Chixiong Huang 1,* , Xinzhong Li 1 , Xiang He 1 , Yanmei Chen 1 , Wangjun Liao 3 , Yulin Liao 1 , Jie Sun 1,4 , Ze Liu 2 , Lintao Zhong 1 and Jianping Bin 1 1 Department of Cardiology, State Key Laboratory of Organ Failure Research, Nanfang Hospital, Southern Medical University, Guangzhou, China 2 Wards of Cadres, Guangzhou General Hospital of Guangzhou Military Region, Guangzhou, China 3 Department of Oncology, Nanfang Hospital, Southern Medical University, Guangzhou, China 4 Department of Cardiology, Zhongshan Hospital, Sun Yat-Sen University, Zhongshan, China * These authors have contributed equally to this work Correspondence to: Jianping Bin, email: // Lintao Zhong, email: // Keywords : thoracic aortic dissection, circular RNA, microarray, biomathematics, Pathology Section Received : February 12, 2017 Accepted : June 18, 2017 Published : July 05, 2017 Abstract Objective: To assess the circular RNAs (circRNAs) expression profile and explore the potential functions in human thoracic aortic dissection (TAD). Methods: The differentially expressed circRNAs profiles of the aortic segments between human type A TAD patients ( n =3) and age-matched normal donors (NA; n =3) were analyzed using the Arraystar human circRNAs microarray. Quantitative real-time PCR was used to validate the expression pattern of circRNAs, parental genes, and hsa-miR-320a; Western blotting confirmed MMP9 expression with additional samples. Bioinformatic tools including network analysis, Gene ontology, and KEGG pathway analysis were utilized. Results: Among 8,173 detected circRNA genes, 156 upregulated and 106 downregulated significantly in human TAD as compared to NA (P£0.05). Quantitative real-time PCR showed an elevated expression of the upregulated hsa_circRNA_101238, hsa_circRNA_104634, hsa_circRNA_002271, hsa_circRNA_102771, hsa_circRNA_104349, COL1A1, and COL6A3 and reduced of the downregulated hsa_circRNA_102683, hsa_circRNA_005525, hsa_circRNA_103458, and FLNA. Gene ontology analysis revealed that the parental genes favored several pathological processes, such as negative regulation of cell proliferation and extracellular matrix organization. The circRNA-miRNA co-expression network predicted that 33 circRNAs might interact with at least one target miRNAs altered in TAD. KEGG pathway analysis revealed that 28 altered miRNAs were enriched on focal adhesion and vascular smooth muscle contraction. The hsa_circRNA_101238-miRNA-mRNA network indicated the highest degree of hsa-miR-320a. Quantitative real-time PCR and Western blot manifested the low expression of hsa-miR-320a and high of MMP9 in human TAD tissues, respectively. Conclusions: This study revealed hundreds of differentially expressed circular RNAs in human TAD, suggesting that hsa_circRNA_101238 might inhibit the expression of hsa-miR-320a and increase that of MMP9 in TAD.

// Attila Kovecsi 1,2,* , Ioan Jung 1,* , Zoltan Szentirmay 3 , Tivadar Bara 4 , Tivadar Bara Jr 4 , Daniel Popa 4 and Simona Gurzu 1,2,5 1 Department of Pathology, University of Medicine and Pharmacy, Tirgu Mures, Romania 2 Department of Pathology, Clinical County Emergency Hospital, Tirgu Mures, Romania 3 Department of Molecular Pathology, National Institute of Onology, Budapest, Hungary 4 Department of Surgery, University of Medicine and Pharmacy, Tirgu Mures, Romania 5 Department of Pathology, CCAMF-Research Center, Tirgu Mures, Romania * These authors have equal contribution to the paper Correspondence to: Simona Gurzu, email: // Keywords : c-theta, protein kinase, GIST, diagnosis, CD117, Pathology Section Received : April 04, 2017 Accepted : June 04, 2017 Published : July 08, 2017 Abstract Background: The aim of this study was to evaluate the diagnosis value of an immunohistochemical (IHC) panel of three antibodies for the diagnosis of gastrointestinal stromal tumors (GISTs). Material and methods: In 80 consecutive GISTs without lymph node metastases, the IHC examinations were performed using the antibodies CD117 (c-KIT), DOG-1 and c-theta (PKCθ) protein. The diagnostic value of PKCθ in c-KIT/DOG-1 negative GISTs has been explored in fewer than 10 Medline-indexed papers. Results: The c-KIT, PKCθ and DOG-1 positivity was noted in 92.50% ( n = 74), 90% ( n = 72) and 76.25% ( n = 61) of the cases, respectively. All of the C-KIT negative cases ( n = 6) were also DOG-1 negative but displayed PKCθ positivity. All of the DOG-1 positive cases ( n = 61) also expressed c-KIT. No correlation between the examined markers and clinicopathological parameters was noted. Conclusions: The PKCθ sensitivity is similar to c-KIT and superior to DOG-1 sensitivity. All of the c-KIT/DOG-1 negative GISTs seem to express PKCθ. For a proper diagnosis of GIST, the c-KIT/DOG-1/PKCθ panel should be used, with possible therapeutic but not prognostic value.

// Tao Li 1 , Min Shan 1,2 , Jing He 1 , Xiaolan Wang 1 , Shaohui Wang 1 , Mingxing Tian 1 , Jingjing Qi 1 , Tingrong Luo 2 , Yonghong Shi 1 , Chan Ding 1 and Shengqing Yu 1 1 Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China 2 College of Animal Science and Technology, Guangxi University, Guangxi, China Correspondence to: Shengqing Yu, email: // Keywords : Riemerella anatipestifer , M949_0459 gene, MIC, tigecycline, resistance, Pathology Section Received : May 16, 2017 Accepted : June 29, 2017 Published : July 27, 2017 Abstract Based on its important role in last-line therapeutics against multidrug-resistant bacteria, tigecycline has been increasingly important in treating infections. However, mounting reports on tigecycline-resistant bacterial strains isolated from different sources are of concern, and molecular mechanisms regarding tigecycline resistance are poorly understood. Riemerella anatipestifer is a Gram-negative, non-motile, non-spore-forming, rod-shaped bacterium, which causes fibrinous pericarditis, perihepatitis, and meningitis in infected ducks. We previously constructed a random transposon mutant library using Riemerella anatipestifer strain CH3, in present study, we described that Riemerella anatipestifer M949_0459 gene is responsible for the bacterial resistance to tigecycline. Using the minimum inhibitory concentration assay, a mutant strain showed significantly increased (about six-fold) tigecycline susceptibility. Subsequently, the knocked-down gene was identified as M949_0459 , a putative flavin adenine dinucleotide-dependent oxidoreductase. To confirm the resistance function, M949_0459 gene was overexpressed in Escherichia coli strain BL21, and the minimum inhibitory concentration analysis showed that the gene product conferred resistance to tigecycline. Additionally, expression of the M949_0459 gene under treatment with tigecycline was measured with quantitative real-time PCR. Results showed that the mRNA expression of M949_0459 gene was elevated under tigecycline treatment with dose range of 1-10 mg/L, and peaked at 4 mg/L. Moreover, two kinds of efflux pump inhibitors, carbonyl cyanide m-chlorophenyl hydrazone and phenylalanine arginyl β -naphthylamide were tested, which showed no function on tigecycline resistance in the strain CH3. Our results may provide insights into molecular mechanisms for chemotherapy in combating Riemerella anatipestifer infections.

// Guiju Tang 1 , Zhihong Weng 2 , Jun Song 3 and Yixiong Chen 3 1 Department of Obstetrics and Gynecology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China 2 Department of Infectious Disease, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China 3 Division of Gastroenterology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China Correspondence to: Yixiong Chen, email: // Keywords : recombinant adeno-associated virus, Jagged1, hepatic fibrosis, hepatic stellate cells, epithelial-mesenchymal transition, Pathology Section Received : November 30, 2016 Accepted : June 02, 2017 Published : June 15, 2017 Abstract The role of the Notch ligand Jagged1 in hepatic fibrosis remains to be elucidated. In the current study, we investigated the role of Jagged1 in the activation of hepatic stellate cells (HSCs) and development of hepatic fibrosis in rats. In vitro , Jagged1 in HSCs was downregulated and upregulated by Jagged1 siRNA and pcDNA3.1 Jagged1, respectively. The levels of epithelial-mesenchymal transition (EMT) markers and HSC activation markers were assessed using western blot analysis. The proliferation and migration capacity of HSCs were assessed using 5-ethynyl-2′-deoxyuridine (EdU) incorporation and Transwell migration assays. In vivo , a recombinant adeno-associated virus type 1 (rAAV1) vector carrying Jagged1 shRNA (rAAV1-Jagged1-shRNA) was constructed and transferred to rat livers via the tail vein. Reversion of liver fibrosis and the effect of Jagged1 signaling on EMT were studied using pathological, immunohistochemical and immunofluorescence methods. Our findings revealed that downregulation and upregulation of Jagged1 inhibited and promoted, respectively, HSC activation. The migratory capacity of HSCs was markedly restrained by Jagged1 siRNA. Furthermore, downregulation of Jagged1 suppressed EMT in HSCs. rAAV1-Jagged1-shRNA was generated to treat CCl4-induced hepatic fibrosis in rats. Treatment with rAAV1-Jagged1-shRNA reversed hepatic fibrosis by decreasing EMT. The results of the present study suggest that inhibition of Jagged1 is a potential treatment to ameliorate liver fibrosis.

Lung injury is one of the pathological features in human or animal after radiation and the main side effect for patient after lung cancer radiotherapy. The efficient protective strategy still needs to exploit and the underlying mechanisms remain to be investigated. We found that the expression and activity of matrix metalloproteinases (MMPs) significantly increased at the early stage of radiation-induced lung injury (RILI). Pretreatment with Ilomastat, a synthetic inhibitor of MMPs, decreased the expression and activity of MMPs and significantly alleviated the lung inflammation and fibrosis in the irradiated mice, as well as enhanced the survival of irradiated mice. In addition, the levels of TGF-β, IL-6, TNF-α and IL-1β in the tissues dramatically reduced in the irradiated mice pretreated with Ilomastat. Furthermore, our experiments in vitro also showed that radiation significantly increased the MMPs activity, and Ilomastat pretreatment inhibited the activity of MMPs activated by irradiation and increased the cell survival. It is the first report, to our knowledge, to demonstrate that Ilomastat is a potential effective reliever for RILI and MMPs may play important roles in the process of RILI.

// Olga A. Efimova 1,2,* , Anna A. Pendina 1,2,3,* , Andrei V. Tikhonov 1,2,3,* , Sergey E. Parfenyev 2 , Irina D. Mekina 1 , Evgeniia M. Komarova 1 , Mariia A. Mazilina 1,2 , Eugene V. Daev 2 , Olga G. Chiryaeva 1,4,5 , Ilona A. Galembo 3 , Mikhail I. Krapivin 2 , Oleg S. Glotov 1,2 , Irina S. Stepanova 6,7 , Svetlana A. Shlykova 8 , Igor Yu. Kogan 1 , Alexander M. Gzgzyan 1,2 , Tatyana V. Kuznetzova 1,2 and Vladislav S. Baranov 1,2 1 D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology, St. Petersburg, Russia 2 St. Petersburg State University, St. Petersburg, Russia 3 Center for Medical Genetics, St. Petersburg, Russia 4 St. Petersburg State Pediatric Medical University, St. Petersburg, Russia 5 S.M. Kirov Military Medical Academy, St. Petersburg, Russia 6 Institute of Cytology RAS, St. Petersburg, Russia 7 Aimed Clinic, St. Petersburg, Russia 8 AVA-Peter Clinic, St. Petersburg, Russia * These authors have contributed equally to this work Correspondence to: Olga A. Efimova, email: // Keywords : 5-hydroxymethylcytosine, semen quality, sperm DNA fragmentation, human spermatogenesis, testicular spermatogenic cells, Pathology Section Received : August 01, 2016 Accepted : May 21, 2017 Published : June 01, 2017 Abstract We performed immunofluorescent analysis of DNA hydroxymethylation and methylation in human testicular spermatogenic cells from azoospermic patients and ejaculated spermatozoa from sperm donors and patients from infertile couples. In contrast to methylation which was present throughout spermatogenesis, hydroxymethylation was either high or almost undetectable in both spermatogenic cells and ejaculated spermatozoa. On testicular cytogenetic preparations, 5-hydroxymethylcytosine was undetectable in mitotic and meiotic chromosomes, and was present exclusively in interphase spermatogonia Ad and in a minor spermatid population. The proportions of hydroxymethylated and non-hydroxymethylated diploid and haploid nuclei were similar among samples, suggesting that the observed alterations of 5-hydroxymethylcytosine patterns in differentiating spermatogenic cells are programmed. In ejaculates, a few spermatozoa had high 5-hydroxymethylcytosine level, while in the other ones hydroxymethylation was almost undetectable. The percentage of highly hydroxymethylated (5-hydroxymethylcytosine-positive) spermatozoa varied strongly among individuals. In patients from infertile couples, it was higher than in sperm donors ( P

// Julia Onken 1,2 , Peter Vajkoczy 1 , Robert Torka 3 , Claudia Hempt 1 , Victor Patsouris 1 , Frank L. Heppner 2,4,5,6 and Josefine Radke 2,4,6 1 Department of Neurosurgery, Charite - Universitatsmedizin Berlin, Berlin, Germany 2 Berlin Institute of Health (BIH), Berlin, Germany 3 Institute of Physiological Chemistry, Martin Luther University of Halle-Wittenberg, Halle/Saale, Germany 4 Department of Neuropathology, Charite - Universitatsmedizin Berlin, Berlin, Germany 5 Cluster of Excellence, NeuroCure, Berlin, Germany 6 German Cancer Consortium (DKTK), Heidelberg, Germany, Partner Site Charite Berlin, Berlin, Germany Correspondence to: Peter Vajkoczy, email: // Keywords : glioblastoma multiforme (GBM), receptor tyrosine kinase AXL (AXL), glomeroid tufts, overall survival, phospho-Axl (P-AXL), Pathology Section Received : March 23, 2017 Accepted : May 10, 2017 Published : June 13, 2017 Abstract Receptor tyrosine kinase AXL (RTK-AXL) is regarded as a suitable target in glioblastoma (GBM) therapy. Since AXL kinase inhibitors are about to get approval for clinical use, patients with a potential benefit from therapy targeting AXL need to be identified. We therefore assessed the expression pattern of Phospho-AXL (P-AXL), the biologically active form of AXL, in 90 patients with newly diagnosed GBM, which was found to be detectable in 67 patients (corresponding to 74%). We identified three main P-AXL expression patterns: i) exclusively in the tumor vasculature (13%), ii) in areas of hypercellularity (35%), or iii) both, in the tumor vasculature and in hypercellular areas of the tumor tissue (52%). Pattern iii) is associated with significant decrease in overall survival (Hazard ratio 2.349, 95% confidence interval 1.069 to 5.162, * p =0.03). Our data suggest that P-AXL may serve as a therapeutic target in the majority of GBM patients.

Liver fibrosis, leading to cirrhosis and liver failure, can occur after chronic liver injury. The transition of hepatic stellate cells (HSCs) from quiescent cells into proliferative and fibrogenic cells is a central event in liver fibrosis. Here, we show that RAS protein activator like-1 (RASAL1), a RAS-GTPase-activating protein, which switches off RAS activity, is significantly decreased during HSC activation, and that HSC activation can be antagonized by forced expression of the RASAL1 protein. We demonstrate that RASAL1 suppresses HSC proliferation by regulating the Ras-MAPK pathway, and that RASAL1 suppresses HSC fibrogenic activity by regulating the PKA-LKB1-AMPK-SRF pathway by interacting with angiotensin II receptor, type 1. We also show that RASAL1-deficient mice are more susceptible to liver fibrosis. These data demonstrate that deregulated RASAL1 expression levels and the affected downstream intracellular signaling are central mediators of perpetuated HSC activation and fibrogenesis in the liver.

// Dong Han 1,2 , Xiang Li 2 , Wen-Si Fan 2 , Jiang-Wei Chen 2 , Tian-Tian Gou 2 , Tao Su 2 , Miao-Miao Fan 2 , Meng-Qi Xu 1 , Ya-Bin Wang 1 , Sai Ma 2 , Ya Qiu 1 and Feng Cao 1,2 1 Department of Cardiology, State Key Laboratory of Kidney Diseases, Chinese PLA General Hospital, Beijing, China 2 Department of Cardiology, Xijing Hospital, Fourth Military Medical University, Xi’an, Shaanxi, China Correspondence to: Feng Cao, email: // Keywords : cannabinoid receptor type II, stem cells, oxidative stress, myocardial infarction, Stat3, Pathology Section Received : February 12, 2017 Accepted : April 21, 2017 Published : May 04, 2017 Abstract The poor survival of cells in ischemic sites diminishes the therapeutic efficacy of stem cell therapy. Previously we and others have reported that Cannabinoid receptor type II (CB2) is protective during heart ischemic injury for its anti-oxidative activity. However, whether CB2 activation could improve the survival and therapeutic efficacy of stem cells in ischemic myocardium and the underlying mechanisms remain elusive. Here, we showed evidence that CB2 agonist AM1241 treatment could improve the functional survival of adipose-derived mesenchymal stem cells (AD-MSCs) in vitro as well as in vivo . Moreover, AD-MSCs adjuvant with AM1241 improved cardiac function, and inhibited cardiac oxidative stress, apoptosis and fibrosis. To unveil possible mechanisms, AD-MSCs were exposed to hydrogen peroxide/serum deprivation to simulate the ischemic environmentin myocardium. Results delineated that AM1241 blocked the apoptosis, oxidative damage and promoted the paracrine effects of AD-MSCs. Mechanistically, AM1241 activated signal transducers and activators of transcription 3 (Stat3) through the phosphorylation of Akt and ERK1/2. Moreover, the administration of AM630, LY294002, U0126 and AG490 (inhibitors for CB2, Akt, ERK1/2 and Stat3, respectively) could abolish the beneficial actions of AM1241. Our result support the promise of CB2 activation as an effective strategy to optimize stem cell-based therapy possibly through Stat3 activation.

// Yuning Song 1,* , Tingjun Liu 1,* , Yong Wang 1 , Jichao Deng 1 , Mao Chen 1 , Lin Yuan 1 , Yi Lu 1 , Yuxin Xu 1 , Haobin Yao 1 , Zhanjun Li 1 and Liangxue Lai 1,2 1 Jilin Provincial Key Laboratory of Animal Embryo Engineering, Jilin University, Changchun, China 2 Key Laboratory of Regenerative Biology, and Guangdong Provincial Key Laboratory of Stem Cells and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, Guangdong, China * These authors have contributed equally to this work Correspondence to: Zhanjun Li, email: // Liangxue Lai, email: // Keywords : Sp1; SRY; CRISPR/Cas9; sex reversed; Pathology Section Received : January 25, 2017 Accepted : March 28, 2017 Published : April 09, 2017 Abstract Sex-determining region Y is a crucial gene that initiates male sex determination in mammals. Mutations of the Sp1-binding site in the 5’ flanking region of SRY are associated with clinical male-to-female sex reversal syndrome, although such occurrences are rare and, until now, have not been reported in animal models. In this study, we mutated Sp1-binding sites in the 5’ flanking region of the rabbit SRY gene using the CRISPR/Cas9 system. As expected, the SRY -Sp1 knockout rabbits had female external and internal genitalia and exhibited normal female copulatory behaviors, but they were infertile, and the adults displayed reduced follicles. Interestingly, we successfully obtained offspring from sex-reversed SRY -Sp1 knockout rabbits using embryo transfer. In summary, our study demonstrates that Sp1 is a major regulator in SRY gene transcription, and mutations of the Sp1 binding sites (Sp1-B and Sp1-C) in the 5’ flanking region of SRY induce sex reversal in rabbits, which can be used as targets for clinical research of male-to-female sex reversal syndrome. Additionally, we provide the first evidence that sex reversal syndrome patients have the potential to become pregnant with the use of embryo transfer.

Obesity is characterized by increased fat mass, as adipose tissue serves as a storage site for excess energy from food consumption. In obesity, altered lipid metabolism of adipose tissue, characterized by fatty acid uptake, de novo lipogenesis, and lipolysis, are induced. In this study, we examined the effect of zerumbone, a major sesquiterpene from wild ginger, on high-fat diet (HF)-induced obesity and dysregulated lipid metabolism in the white adipose tissues (WAT) of C57BL/6N mice. Dietary supplementation with zerumbone ameliorated HF-induced obesity and improved impaired lipid metabolism in WAT. Zerumbone additionally induced AMPK activation and phosphorylation of acetyl-CoA carboxylase, and effectively decreased adipogenic differentiation, in a concentration-dependent manner in the 3T3-L1 cells. Dysregulated microRNAs in obese WAT and adipocytes were examined, and zerumbone treatment was found to effectively reverse the robust upregulation of microRNA-146b. An increase in the levels of SIRT1, the direct target of microRNA-146b, was observed in zerumbone-treated differentiated adipocytes. This increase was additionally observed in WAT of zerumbone-supplemented mice. The antiadipogenic effect of zerumbone was found to be abolished in SIRT1-silenced 3T3-L1 cells. The increase in SIRT1 levels induced by zerumbone led to deacetylation of FOXO1 and PGC1α in WAT and differentiated 3T3-L1 cells. These findings indicate that zerumbone ameliorated diet-induced obesity and inhibited adipogenesis, and that the underlying mechanisms involved AMPK and the microRNA-146b/SIRT1 pathway. Zerumbone may represent a potential therapeutic candidate for the prevention and treatment of metabolic diseases, particularly obesity.

Killer cell immunoglobulin-like receptor (KIR) 2DL4 (CD158d) is a receptor for human leukocyte antigen-G. The function of KIR2DL4 has been reported in human natural killer cell lymphoma and mastocytosis, but not in Langerhans cell histiocytosis (LCH). Herein, we examined the expression and function of KIR2DL4 in LCHs. In pathological specimens, 27 of 36 LCH cases (75.0%) were immunohistochemically positive for KIR2DL4. Its expression was independent of age, gender, location, multi- or single-system, and the status of BRAFV600E immunostaining. We also confirmed the expression of KIR2DL4 mRNA and protein in the human LCH-like cell lines ELD-1 and PRU-1. KIR2DL4 protein was distributed in the membrane and cytoplasm of ELD-1 cells, but only in the cytoplasm of PRU-1 cells. An agonistic antibody against KIR2DL4 reduced phosphorylation of extracellular signal-regulated kinases (ERKs) and suppressed the cell growth of ELD-1 cells in a Src homology region 2 domain-containing phosphatase-2 dependent manner, but it had no effect in PRU-1 cells. These results suggest that KIR2DL4-mediated ERK suppression is a possible therapeutic target for LCH cells.

// Moises Freitas-Andrade 1,2 , John F. Bechberger 1 , Brian A. MacVicar 2 , Victor Viau 1 and Christian C. Naus 1 1 Department of Cellular and Physiological Sciences, The Life Science Institute, University of British Columbia, Vancouver, British Columbia, Canada 2 Department of Psychiatry, Djavad Mowafaghian Centre for Brain Health, University of British Columbia, Vancouver, British Columbia, Canada Correspondence to: Christian C. Naus, email: // Keywords : stroke, neuroprotection, neuroinflammation, sex-differences, pannexin, Pathology Section Received : December 15, 2016 Accepted : March 22, 2017 Published : April 07, 2017 Abstract The membrane channel Pannexin 1 (Panx1) mediates apoptotic and inflammatory signaling cascades in injured neurons, responses previously shown to be sexually dimorphic under ischemic conditions. We tested the hypothesis that Panx1 plays an underlying role in mediating sex differences in stroke outcome responses. Middle-aged, 8-9 month old male and female wild type and Panx1 KO mice were subjected to permanent middle cerebral artery (MCA) occlusion, and infarct size and astrocyte and microglia activation were assessed 4 days later. The sexually dimorphic nature of Panx1 deletion was also explored by testing the effect of probenecid a known Panx1 blocker to alter stroke volume. Panx1 KO females displayed significantly smaller infarct volumes (~ 50 % reduction) compared to their wild-type counterparts, whereas no such KO effect occurred in males. This sex-specific effect of Panx1 KO was recapitulated by significant reductions in peri-infarct inflammation and astrocyte reactivity, as well as smaller infarct volumes in probenecid treated females, but not males. Finally, females showed overall, higher Panx1 protein levels than males under ischemic conditions. These findings unmask a deleterious role for Panx1 in response to permanent MCA occlusion, that is unique to females, and provide several new frameworks for understanding sex differences in stroke outcome.

// Joshua Kingman 1 , Jouni Uitto 1 and Qiaoli Li 1 1 Department of Dermatology and Cutaneous Biology, The Sidney Kimmel Medical College, and the PXE International Center of Excellence in Research and Clinical Care, Thomas Jefferson University, Philadelphia, PA, USA Correspondence to: Qiaoli Li, email: // Keywords : mouse model, ectopic mineralization, maternal diet, magnesium, generalized arterial calcification of infancy, Pathology Section Received : October 28, 2016 Accepted : March 16, 2017 Published : March 29, 2017 Abstract Generalized arterial calcification of infancy (GACI) is an autosomal recessive disorder caused by mutations in the ENPP1 gene. It is characterized by mineralization of the arterial blood vessels, often diagnosed prenatally, and associated with death in early childhood. There is no effective treatment for this devastating disorder. We previously characterized the Enpp1 asj mutant mouse as a model of GACI, and we have now explored the effect of elevated dietary magnesium (five-fold) in pregnant mothers and continuing for the first 14 weeks of postnatal life. The mothers were kept on either control diet or experimental diet supplemented with magnesium. Upon weaning at 4 weeks of age the pups were placed either on control diet or high magnesium diet. The degree of mineralization was assessed at 14 weeks of age by histopathology and a chemical calcium assay in muzzle skin, kidney and aorta. Mice placed on high magnesium diet showed little, if any, evidence of mineralization when their corresponding mothers were also placed on diet enriched with magnesium during pregnancy and nursing. The reduced ectopic mineralization in these mice was accompanied by increased calcium and magnesium content in the urine, suggesting that magnesium competes calcium-phosphate binding thereby preventing the mineral deposition. These results have implications for dietary management of pregnancies in which the fetus is suspected of having GACI. Moreover, augmenting a diet with high magnesium may be beneficial for other ectopic mineralization diseases, including nephrocalcinosis.

// Da Xie 1,* , Yong Xu 2,* , Wang Jing 3,* , Zeng Juxiang 4 , Li Hailun 2 , Hu Yu 2 , Dong-Hui Zheng 2 and Yong-Tao Lin 4 1 Department of Nephrology, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, China 2 Department of Nephrology, Huai’an Hospital Affiliated to Xuzhou Medical University and Huai’an Second Hospital, Huai’an, China 3 Department of Pediatrics, Huai’an Hospital Affiliated to Xuzhou Medical University and Huai’an Second Hospital, Huai’an, China 4 Jiangsu College of Nursing, Huai’an, Jiangsu, China * Co-first authors Correspondence to: Yong-Tao Lin, email: // Dong-Hui Zheng, email: // Keywords : renal ischemia-reperfusion; berberine; nanoparticles; oxidative stress; apoptosis; Pathology Section Received : December 09, 2016 Accepted : February 13, 2017 Published : March 23, 2017 Abstract Renal ischemia-reperfusion (I/R) injury is one of the most common causes of acute renal failure, the prognosis of which remains poor and there still lacks of effective therapeutics available in the clinic. This study aimed at investigating the effects of Berberine nanoparticles (BBR-NP) on the ischemia-reperfusion injury of renal tubular epithelial cells and underlying the mechanisms. Our results showed that in a rat model of renal I/R injury, BBR and BBR-NP protected renal against injury both functionally (as assessed by serum urea nitrogen and creatinine level) and morphologically (as assessed by HE staining, transmission electron microscopy and TUNEL staining) in a dose-dependent manner, with the effects of BBR-NP superior to BBR alone. Mechanism investigation showed that BBR-NP reversed oxidative stress and subsequent apoptosis of renal cells, as demonstrated by the decreased expression of proteins involved in the oxidative stress and mitochondrial stress pathways. In conclusion, our study showed that BBR-NP is superior to BBR alone in protecting renal against I/R injury and explored the underlying mechanisms, which should be tested in further studies and might give impetus to the development of novel therapeutics based on BBR-NP against renal I/R.

We evaluated the frequency of translocation renal cell carcinoma (RCC) by reverse transcription polymerase chain reaction (RT-PCR) and how well the TFE3 immunoreactivity is concordant with TFE3 gene translocation status proved by fluorescence in situ hybridization (FISH) assay and RT-PCR. TFE3 and Cathepsin K expression was analyzed by immunohistochemistry in 185 RCC cases, and 48 cases either of more than weak expression of TFE3 or of positivity for Cathepsin K were done for FISH analysis and RT-PCR. All the RT-PCR positive cases were confirmed by cloning and sequencing. Of the 14 cases with strong nuclear TFE3 expression, 12 showed a break-apart signal by FISH. ASPL- and PRCC-TFE3 translocations were detected in 13 and one case, respectively, by RT-PCR. Of 21 cases with weak TFE3 expression, five were translocation-positive by FISH. ASPL-, PRCC-, and PSF-TFE3 translocations were detected by RT-PCR (n=3, 3, and 1, respectively). All 13 TFE3-negative/cathepsin K-positive cases were negative by FISH and two each harbored ASPL- and PRCC-TFE3 translocations that were detected by RT-PCR. A high rate of TFE3 immunoreactivity (8.6%) was confirmed by RT-PCR (13.5%) and FISH (9.7%). Higher translocation rate of RT-PCR means RT-PCR detected translocation in TFE3 weak expression group and only cathepsin K positive group more specifically than FISH. Thus, RT-PCR would complement FISH analysis for detecting translocation RCC with fusion partners.

Metastasis is a major cause of mortality for cancer patients and remains as the greatest challenge in cancer therapy. Driven by multiple factors, metastasis may not be controlled by the inhibition of single target. This study was aimed at assessing the hypothesis that drugs could be rationally combined to co-target critical DNA, RNA and protein molecules to achieve "saturation attack" against metastasis. Independent actions of the model drugs DNA-intercalating doxorubicin, RNA-interfering miR-34a and protein-inhibiting sorafenib on DNA replication, RNA translation and protein kinase signaling in highly metastatic, human osteosarcoma 143B cells were demonstrated by the increase of γH2A.X foci formation, reduction of c-MET expression and inhibition of Erk1/2 phosphorylation, respectively, and optimal effects were found for triple-drug combination. Consequently, triple-drug treatment showed a strong synergism in suppressing 143B cell proliferation and the greatest effects in reducing cell invasion. Compared to single- and dual-drug treatment, triple-drug therapy suppressed pulmonary metastases and orthotopic osteosarcoma progression to significantly greater degrees in orthotopic osteosarcoma xenograft/spontaneous metastases mouse models, while none showed significant toxicity. In addition, triple-drug therapy improved the overall survival to the greatest extent in experimental metastases mouse models. These findings demonstrate co-targeting of DNA, RNA and protein molecules as a novel therapeutic strategy for the treatment of metastasis.

// Maria Adelaide Marini 1,* , Gaia Chiara Mannino 2,* , Teresa Vanessa Fiorentino 2 , Francesco Andreozzi 2 , Francesco Perticone 2 and Giorgio Sesti 2 1 Department of Systems Medicine, University of Rome Tor Vergata, Rome, Italy 2 Department of Medical and Surgical Sciences, University Magna-Graecia of Catanzaro, Catanzaro, Italy * These authors have equally contributed as first authors to the work Correspondence to: Giorgio Sesti, email: // Keywords : insulin-like growth factor 1, hemoglobin, rs35767, anemia, single nucleotide polymorphism, Pathology Section Received : January 30, 2017 Accepted : March 02, 2017 Published : March 11, 2017 Abstract In vitro and in vivo studies suggest that IGF-1 has a role in erythropoiesis. There is evidence that the rs35767 C/T polymorphism near IGF1 is associated with plasma IGF-1 levels. We investigated the effect of this polymorphism on hemoglobin (Hb) concentration and anemia. The study group comprised 3286 adult Whites. The rs35767 polymorphism was screened using a TaqMan allelic discrimination assay. The rs35767 polymorphism was not associated with age, gender, BMI, waist circumference, smoking, blood pressure, plasma glucose, HbA1c, type 2 diabetes, HOMA-IR, hsCRP, eGFR, and lipid profile. Erythrocyte sedimentation rate (ESR), fibrinogen, and fasting insulin levels were significantly lower in TT genotype carriers compared with C allele carriers. Hb concentration was significantly higher in carriers of the TT genotype compared with C allele carriers, and a lower proportion of TT carriers had anemia. As compared with TT genotype carriers, those bearing the CC genotype had a 2.4-fold higher risk of anemia (OR 2.40, 95%CI 1.19-4.82), and those with the CT genotype had a 2.0-fold higher risk of anemia (OR 2.06, 95%CI 1.04-4.11). The association remained significant when fasting insulin, eGFR, smoking, diabetes, ACE inhibitors, sartans or diuretics treatments, use of metformin and pioglitazone were added to the model, but its independence was not retained after inclusion of fibrinogen and ESR values into the model. In conclusion, rs35767 TT allele carriers exhibited significantly higher concentrations of Hb, and lower risk of anemia compared with C allele carriers supporting the idea that IGF-1 plays a role in erythropoiesis homeostasis.

// Simone Muenst 1,* , Robert Mechera 2,3,* , Silvio Daster 2 , Salvatore Piscuoglio 1 , Charlotte K.Y. Ng 1 , Fabienne Meier-Abt 4 , Walter P. Weber 2,* and Savas D. Soysal 2,* 1 Institute of Pathology, University Hospital Basel, Basel, Switzerland 2 Department of Surgery, University Hospital Basel, Basel, Switzerland 3 Department of Biomedicine, University Hospital Basel, Basel, Switzerland 4 Institute of Hematology, University Hospital Zurich, Zurich, Switzerland * These authors have contributed equally to this work Correspondence to: Savas D. Soysal, email: // Keywords : early pregnancy, breast cancer, Wnt signaling pathway, breast cancer prevention, Pathology Section Received : November 18, 2016 Accepted : February 22, 2017 Published : March 08, 2017 Abstract Background: Pregnancy at early age is the most significant modifiable factor which consistently decreases lifetime breast cancer risk. However, the underlying mechanisms haven’t been conclusively identified. Studies in mice suggest a reduction in progesterone-receptor (PR) sensitive epithelial cells as well as a downregulation of the Wnt signaling pathway as being one of the main mechanisms for the protective effect of early pregnancy. The aim of our study was to validate these findings in humans. Methods: We collected benign breast tissue of 125 women who had been stratified according to age at first pregnancy and the occurrence of subsequent breast cancer, and performed immunohistochemistry for PR, Wnt4 and the Wnt-target Versican. Results: The number of PR positive epithelial cells was significantly lower in the group of women with early pregnancy and no subsequent breast cancer compared to the group of nulliparous women with subsequent invasive breast cancer ( p = 0.0135). In women with early pregnancy, expression of Versican and Wnt4 was significantly lower compared to nulliparous women ( p = 0.0036 and p = 0.0241 respectively), and Versican expression was also significant lower compared to women with late pregnancy ( p < 0.0001). Discussion: Our results confirm prior observations in mice and suggest a role of downregulation of epithelial Wnt signaling in the protective effect of early pregnancy in humans. This results in a decreased proliferation of stem/progenitor cells; therefore, the Wnt signaling pathway may represent a potential target for breast cancer prevention in humans.

// Andrea Cerasuolo 1 , Clorinda Annunziata 1 , Marianna Tortora 1 , Noemy Starita 1 , Giovanni Stellato 2 , Stefano Greggi 2 , Maria Grazia Maglione 3 , Franco Ionna 3 , Simona Losito 4 , Gerardo Botti 4 , Luigi Buonaguro 1 , Franco M. Buonaguro 1 and Maria Lina Tornesello 1 1 Molecular Biology and Viral Oncology Unit, Istituto Nazionale Tumori IRCCS “Fondazione G. Pascale”, Napoli, Italy 2 Gynecology Oncology Division, Istituto Nazionale Tumori IRCCS “Fondazione G. Pascale”, Napoli, Italy 3 Department of Maxillofacial and Ear Nose and Throat Surgery, Istituto Nazionale Tumori IRCCS “Fondazione G. Pascale”, Napoli, Italy 4 Department of Pathology, Istituto Nazionale Tumori IRCCS “Fondazione G. Pascale”, Napoli, Italy Correspondence to: Maria Lina Tornesello, email: // Keywords : HPV16; E6 gene; E7 gene; cervical carcinoma; cervical intraepithelial neoplasia; Pathology Section Received : February 07, 2016 Accepted : February 28, 2017 Published : March 07, 2017 Abstract Human papillomavirus type 16 (HPV16) is the major cause of cervical cancer and of a fraction of oropharyngeal carcinoma. Few studies compared the viral expression profiles in the two types of tumor. We analyzed HPV genotypes and viral load as well as early (E2/E4, E5, E6, E6*I, E6*II, E7) and late (L1 and L2) gene expression of HPV16 in cervical and oropharyngeal cancer biopsies. The study included 28 cervical squamous cell carcinoma (SCC) and ten oropharyngeal SCC, along with pair-matched non-tumor tissues, as well as four oropharynx dysplastic tissues and 112 cervical intraepithelial neoplasia biopsies. Viral load was found higher in cervical SCC (

// Zhao-Yang Li 1,* , Jia Lin 1,* , Feng Sun 1,* , Hui Li 1,2,* , Jun Xia 1,3 , Xue-Nan Li 1,4 , Jing Ge 1,4 , Cong Zhang 1,3 and Jin-Long Li 1,3,4 1 College of Veterinary Medicine, Northeast Agricultural University, Harbin, P. R. China 2 Harbin Sport University, Harbin, P. R. China 3 Department of Heilongjiang for Common Animal Disease Prevention and Treatment, Key Laboratory of the Provincial Education Northeast Agricultural University, Harbin, P. R. China 4 Heilongjiang Key Laboratory for Laboratory Animals and Comparative Medicine, Northeast Agricultural University, Harbin, P. R. China * These authors have contributed equally to this work Correspondence to: Jin-Long Li, email: // Keywords : transport stress; weight loss; heart injury; ionic disorder; ATPase; Pathology Section Received : January 13, 2017 Accepted : February 24, 2017 Published : March 04, 2017 Abstract Transportation is inevitable in the poultry industry, and it can induce stress to chicks in varying degrees, such as mild discomfort, sometimes even death. However, the research about the effects of transport stress on the weight loss and heart injury of chicks is lacking. To elucidate the underlying mechanism of transport stress-induced effects, chicks were transported for 2h, 4h and 8h. The creatinine kinase (CK) activities, the ionic contents, the ATPases activities and the transcription of the ATPase associated subunits in chick heart were detected. The results showed that transport stress increased the weight loss and the CK activity, disturbed the ionic (K + , Ca 2+ , Mg 2+ ) homeostasis and inhibited the ATPase (Na + -K + -ATPase, Ca 2+ -ATPase, Mg 2+ -ATPase and Ca 2+ -Mg 2+ -ATPase) activities, increased the ATP content and downregulated the gene expression levels of the ATPase associated subunits in heart. In conclusion, transport stress disturbed the ionic homeostasis via modulating ion transporting ATPases and the transcriptions of the associated subunits, and ultimately induced weight loss and heart injury in chicks.

// Vinodkumar B. Pillai 1 , Abhinav Kanwal 1 , Yong Hu Fang 2 , Willard W. Sharp 2 , Sadhana Samant 1 , Jack Arbiser 3 and Mahesh P. Gupta 1 1 Department of Surgery, Pritzker School of Medicine, University of Chicago, Chicago, IL, USA 2 Department of Medicine, Pritzker School of Medicine, University of Chicago, Chicago, IL, USA 3 Department of Dermatology, Atlanta Veterans Administration Health Center, Emory University School of Medicine, Atlanta, GA, USA Correspondence to: Mahesh P. Gupta, email: // Keywords : doxorubicin, cardiac toxicity, SIRT3, Cardiac hypertrophy, cancer therapy, Pathology Section Received : September 14, 2016 Accepted : February 27, 2017 Published : March 11, 2017 Abstract Doxorubicin is the chemotherapeutic drug of choice for a wide variety of cancers, and cardiotoxicity is one of the major side effects of doxorubicin treatment. One of the main cellular targets of doxorubicin in the heart is mitochondria. Mitochondrial sirtuin, SIRT3 has been shown to protect against doxorubicin-induced cardiotoxicity. We have recently identified honokiol (HKL) as an activator of SIRT3, which protects the heart from developing pressure overload hypertrophy. Here, we show that HKL-mediated activation of SIRT3 also protects the heart from doxorubicin-induced cardiac damage without compromising the tumor killing potential of doxorubicin. Doxorubicin-induced cardiotoxicity is associated with increased ROS production and consequent fragmentation of mitochondria and cell death. HKL-mediated activation of SIRT3 prevented Doxorubicin induced ROS production, mitochondrial damage and cell death in rat neonatal cardiomyocytes. HKL also promoted mitochondrial fusion. We also show that treatment with HKL blocked doxorubicin-induced cardiac toxicity in mice. This was associated with reduced mitochondrial DNA damage and improved mitochondrial function. Furthermore, treatments of mice, bearing prostrate tumor-xenografts, with HKL and doxorubicin showed inhibition of tumor growth with significantly reduced cardiac toxicity. Our results suggest that HKL-mediated activation of SIRT3 protects the heart from doxorubicin-induced cardiotoxicity and represents a potentially novel adjunct for chemotherapy treatments.

// Jiping Xu 1,* , Haibin Zhong 1,* , Ling Cui 1,* , Qianqian Lan 1 , Lifei Chen 1 , Wenjing He 1 , Yu Wu 1 , Li Jiang 1 , Hui Huang 1 , Xin Zhao 1 , Li Li 1 , Siming Zeng 1 , Min Li 1 and Fan Xu 1 1 Department of Ophthalmology, People’s Hospital of Guangxi Zhuang Autonomous Region, Nanning, Guangxi, People’s Republic of China * These authors have contributed equally to this work Correspondence to: Fan Xu, email: // Min Li, email: // Keywords : Wip1, epiretinal membranes, proliferative diabetic retinopathy, NF-κB, glial cell, Pathology Section Received : January 06, 2017 Accepted : March 11, 2017 Published : March 29, 2017 Abstract Objective: The aims of the present study were to investigate the expression and distribution of Wild-type p53-induced phosphatase 1 (Wip1) in diabetic patients with proliferative diabetic retinopathy (PDR) with epiretinal membranes (ERMs) meanwhile analyze the colocalization of Wip1 and nuclear factor kappa-B (NF-κB) p65 in ERMs. Methods: ERMs samples were collected from patients with PDR (PDR group) or non-diabetic patients with idiopathic epiretinal membranes (iERMs) (control group) during pars plana vitrectomy. Real-Time PCR analysis was carried out to examine the mRNA expression of Wip1 in ERMs. Immunohistochemical analysis and Immunofluorescent analysis were performed to detect the protein expression of Wip1 in ERMs. Double immunofluorescent staining was performed to detect the colocalization of Wip1 and glial fibrillary acidic protein ( GFAP) (retinal glial cells marker), also Wip1 and NF-κB. Results: ERMs were obtained from 17 eyes of 17 patients with PDR (the PDR group) and 9 eyes of 9 nondiabetic patients (the control group) with iERMs. Our results showed high expression levels of Wip1 mRNAs in ERMs after PDR, but low in iERMs. In addition, both immunohistochemistry and immunofluorescence assay showed strong immunoreactivity for Wip1 in PDR ERMs. Furthermore, Wip1 and GFAP were coexpressed in PDR membranes. Finally, the expression of Wip1 was paralleled with NF-κB. Conclusion: These data support the notion that Wip1 contributes to the formation of the ERMs in PDR membranes via NF-κB signaling.

// Mingjing Shao 1,* , Chuanjun Zhuo 2,3,4,* , Ronghuan Jiang 5 , Guangdong Chen 2 , Jianmin Shan 2 , Jing Ping 2 , Hongjun Tian 3 , Lina Wang 3 , Chongguang Lin 2 and Lirong Hu 2 1 National Integrated Traditional and Western Medicine Center for Cardiovascular Disease, China-Japan Friendship Hospital, Beijing, China 2 Department of Psychological Medicine, Wenzhou Seventh People’s Hospital, Wenzhou, China 3 Department of Psychological Medicine, Tianjin Anding Hospital, Tianjin, China 4 Department of Psychological Medicine, Tianjin Anning Hospital, Tianjin, China 5 Department of Psychological Medicine, Chinese People’s Liberation Army General Hospital, Chinese People’s Liberation Army Medical School, Beijing, China * These authors have contributed equally to this work Correspondence to: Lirong Hu, email: // Keywords : hydrogen sulfide; cardiac hypertrophy; oxidative stress; Nrf2; Pathology Section Received : December 12, 2016 Accepted : February 20, 2017 Published : February 27, 2017 Abstract Cardiac hypertrophy is a critical component of phenotype in the failing heart. Recently, increasing evidence has demonstrated that oxidative stress plays an important role in the pathogenesis of myocardial hypertrophy. In the present study, we generated a mouse model of transverse aortic constriction (TAC) to investigate whether hydrogen sulfide (H 2 S) has protective effects against cardiac hypertrophy. Left ventricular structure was analyzed by two-dimensional echocardiography. Oxidative stress was evaluated by measuring malondialdehyde, superoxide dismutase, glutathione peroxidase and reactive oxygen specie in the myocardium. Angiotensin II (Ang-II) was used to induce cardiomyocyte hypertrophy. Neonatal rat cardiomyocytes pretreated with H 2 S donor sodium hydrosulfide prior to Ang-II exposure were used to determine the involvement of Nrf2 and PI3K/Akt pathway in the antioxidant effects of H 2 S. Our findings showed that H 2 S could protect against cardiac hypertrophy by attenuating oxidative stress. The antioxidant roles of H 2 S in myocardial hypertrophy probably depend on the activation of PI3K/Akt signaling, which consequently increases Nrf2 activity and HO-1 and GCLM expression. In summary, H 2 S may exert antioxidant effect on cardiac hypertrophy via PI3K/Akt-dependent activation of Nrf2 pathway.

// Laura Annaratone 1 , Michele Simonetti 2,* , Erik Wernersson 2,* , Caterina Marchio 1,3 , Silvano Garnerone 2 , Maria Stella Scalzo 1 , Magda Bienko 2 , Roberto Chiarle 4,5 , Anna Sapino 1,6 and Nicola Crosetto 2 1 Department of Medical Sciences, University of Turin, Turin, Italy 2 Department of Medical Biochemistry and Biophysics, Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Karolinska Institutet, Stockholm, Sweden 3 Department of Laboratory Medicine, Pathology Unit, Azienda Ospedaliera Citta della Salute e della Scienza di Torino, Turin, Italy 4 Department of Pathology, Boston Children’s Hospital, Harvard Medical School, Boston, Massachusetts, USA 5 Department of Molecular Biotechnology and Health Sciences, University of Turin, Turin, Italy 6 Candiolo Cancer Institute, FPO, IRCCS, Candiolo, Italy * These authors have contributed equally to this work Correspondence to: Nicola Crosetto, email: // Roberto Chiarle, email: // Anna Sapino, email: // Keywords : single-molecule RNA FISH (smFISH); breast cancer; intra-tumor heterogeneity; epidermal growth factor receptor 2 (HER2); estrogen receptor 1 (ER); Pathology Section Received : July 13, 2016 Accepted : February 06, 2017 Published : February 25, 2017 Abstract Intra-tumor heterogeneity is a pervasive property of human cancers that poses a major clinical challenge. Here, we describe the characterization, at the transcriptional level, of the intra-tumor topography of two prominent breast cancer biomarkers and drug targets, epidermal growth factor receptor 2 (HER2) and estrogen receptor 1 (ER) in 49 archival breast cancer samples. We developed a protocol for single-molecule RNA FISH in formalin-fixed, paraffin-embedded tissue sections (FFPE-smFISH), which enabled us to simultaneously detect and perform absolute quantification of HER2 and ER mature transcripts in single cells and multiple tumor regions. We benchmarked our method with standard diagnostic techniques, demonstrating that FFPE-smFISH is able to correctly classify breast cancers into well-established molecular subgroups. By counting transcripts in thousands of single cells, we identified different expression modes and levels of inter-cellular variability. In samples expressing both HER2 and ER, many cells co-expressed both genes, although expression levels were typically uncorrelated. Finally, we applied diversity metrics from the field of ecology to assess the intra-tumor topography of HER2 and ER gene expression, revealing that the spatial distribution of these key biomarkers can vary substantially even among breast cancers of the same subtype. Our results demonstrate that FFPE-smFISH is a reliable diagnostic assay and a powerful method for quantification of intra-tumor transcriptional heterogeneity of selected biomarkers in clinical samples.

// Wei Lu 1,* , Jing Feng 1,2,* , Bo Wen 2 , Kewei Wang 1 and Jin-Hui Wang 1,2,3 1 Qingdao University, School of Pharmacy, Qingdao, Shandong, China 2 State Key lab for Brain and Cognitive Sciences, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China 3 University of Chinese Academy of Sciences, Beijing, China * These authors have contributed to this work equally Correspondence to: Jin-Hui Wang, email: // Kewei Wang, email: // Keywords : Epilepsy, spikes, neuron, synaptic transmission and GABA, Pathology Section Received : January 05, 2017 Accepted : February 13, 2017 Published : February 23, 2017 Abstract Background: Epilepsy, a prevalent neurological disorder, appears self-termination. The endogenous mechanism for seizure self-termination remains to be addressed in order to develop new strategies for epilepsy treatment. We aim to examine the role of activity-induced spontaneous spikes at GABAergic neurons as an endogenous mechanism in the seizure self-termination. Methods and Results: Neuronal spikes were induced by depolarization pulses at cortical GABAergic neurons from temporal lobe epilepsy patients and mice, in which some of these neurons fired activity-induced spontaneous spikes. Neural networks including excitatory and inhibitory neurons were computationally constructed, and their functional properties were based on our studies from whole-cell recordings. With the changes in the portion and excitability of inhibitory neurons that generated activity-induced spontaneous spike, the efficacies to suppress synchronous seizure activity were analyzed, such as its onset time, decay slope and spike frequency. The increases in the proportion and excitability of inhibitory neurons that generated activity-induced spontaneous spikes effectively suppressed seizure activity in neural networks. These factors synergistically strengthened the efficacy of seizure activity suppression. Conclusion: Our study supports a notion that activity-induced spontaneous spikes in GABAergic neurons may be an endogenous mechanism for seizure self-termination. A potential therapeutic strategy for epilepsy is to upregulate the cortical inhibitory neurons that generate activity-induced spontaneous spikes.