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Background Proteasome inhibition demonstrates highly effective impact on multiple myeloma (MM) treatment. Here, we aimed to examine anti-tumor efficiency and underlying mechanisms of a novel well tolerated orally applicable proteasome inhibitor NNU546 and its hydrolyzed pharmacologically active form NNU219. Methods Enzyme activities and inhibition assays was performed to evaluate the effect of NNU219 on proteasome. To explore the anti-MM activity of NNU219 and its related mechanism, multiple in vitro assays such as cell viability, cell cycle and apoptosis assays, Co-immunoprecipitation (Co-IP) and ubiquitination, NF-κB inhibition assay were conducted. Furthermore, in vivo studies were performed to determine the effects of NNU219 on tumors. Results NNU219 showed more selective inhibition to proteasome catalytic subunits and less off-target effect than bortezomib ex vivo. Moreover, intravenous and oral administration of either NNU219 or NNU546 led to more sustained pharmacodynamic inhibitions of proteasome activities compared with bortezomib. Importantly, NNU219 exhibited potential anti-MM activity in both MM cell lines and primary samples in vitro. The anti-MM activity of NNU219 was associated with induction of G2/M-phase arrest, as well as induction of apoptosis via activation of the caspase cascade and endoplasmic reticulum stress response. At well-tolerated doses, significant growth-inhibitory effects of NNU219 and NNU546 were observed in 3 different human MM xenograft mouse models. Furthermore, such observation was even found in the presence of a bone marrow microenvironment. Conclusion Taken together, these findings provided the basis for clinical trial of NNU546 to determine its potential as a candidate for MM treatment.

Glucose‑6‑phosphate dehydrogenase (G6PD) is a cytoplasmic enzyme found in human erythrocytes that provides reduced NADPH for cell metabolism. Glutathione produced by the G6PD pathway can reduce the degree of harm caused by reactive oxygen species such as oxygen‑containing free radicals, peroxides and lipid peroxides. Investigation of G6PD has long focused on hemolysis, jaundice and other diseases caused by defects in its function. However, increased mRNA expression levels of G6PD are predictive of adverse clinical outcomes in cancer patients, including increased drug resistance, migration or proliferation of tumor cells. Mutations in the G6PD gene affect protein expression and activity, and alters the balance of redox states, leading to disease. However, the association between G6PD and tumors is incompletely understood. The aim of the present review was to summarize the current body of knowledge on the role of G6PD in tumor progression and the possible regulatory mechanisms involved. It is hypothesized that G6PD will prove to be of value as a target of cancer treatment in the near future.

Non-alcoholic fatty liver disease (NAFLD) is characterized by increased uptake and accumulation of lipids in hepatocytes. Simple steatosis may progress to non-alcoholic steatohepatitis (NASH) with inflammation, hepatocellular injury and fibrosis. CCN1 is an important matrix protein that regulates cell death and promotes immune cell adhesion and may potentially control this process. The role of CCN1 in NASH remains unclear. We investigated the role of CCN1 in the pathogenesis of steatohepatitis. CCN1 upregulation was found to be closely related with steatosis in patients with NASH, obese mice and a FFA-treated hepatocyte model. Controlling the expression of CCN1 in murine NASH models demonstrated that CCN1 increased the severity of steatosis and inflammation. From the sequence results, we found that fatty acid metabolism genes were primarily involved in the MCD mice overexpressing CCN1 compared to the control. Then, the expression of fatty acid metabolism genes was determined using a custom-designed pathway-focused qPCR-based gene expression array. Expression analysis showed that CCN1 overexpression significantly upregulated the expression of fatty acid metabolism-associated genes. In vitro analysis revealed that CCN1 increased the intracellular TG content, the pro-inflammatory cytokines and the expression level of apoptosis-associated proteins in a steatosis model using murine primary hepatocytes. We identified CCN1 as an important positive regulator in NASH.

Summary Background Heterogeneity in the progression of idiopathic pulmonary fibrosis (IPF) might reflect diversity in underlying pathobiology, and represents a major challenge in the prediction of clinical progression and treatment benefit. Previous studies have found peripheral blood concentrations of several protein biomarkers to be prognostic for overall survival duration in patients with IPF, but these findings have generally not been directly compared and replicated between cohorts. We aimed to use the pivotal trials for pirfenidone to evaluate prognostic and predictive properties of biomarkers across multiple endpoints, and whether they are modulated by pirfenidone treatment. Methods We did post-hoc analyses of test and replication cohorts from CAPACITY 004 ( NCT00287716 ), CAPACITY 006 ( NCT00287729 ), and ASCEND ( NCT01366209 ) trials for the plasma proteins CCL13, CCL17, CCL18, CXCL13, CXCL14, COMP, interleukin 13, MMP3, MMP7, osteopontin, periostin, and YKL40. Eligible participants had IPF and received pirfenidone 2403 mg/day or placebo in CAPACITY (test cohort) or ASCEND (replication cohort), were aged 40–80 years, and without missing biomarker data at baseline. To identify biomarkers that were consistently prognostic for clinical outcome measures, the primary analysis was the association between biomarker concentrations at baseline and absolute change in percentage of predicted forced vital capacity (FVC%pred) at 12 months (CAPACITY week 48, ASCEND week 52) in the placebo group. Biomarkers within the test cohort that met predefined success criteria of a prognostic p value less than 0·10 from multivariate analysis were further assessed in the replication cohort. Furthermore, the predictive effect size (ie, biomarkers that were predictive for benefit from pirfenidone) was calculated as the difference in FVC%pred treatment effect (pirfenidone in relation to placebo) between high versus low biomarker subgroups at week 48 (test cohort) or week 52 (replication cohort). Findings Several baseline biomarkers (CCL13, CCL18, COMP, CXCL13, CXCL14, periostin, and YKL40) were prognostic for progression outcomes in the placebo groups of the test cohort. However, only CCL18 was consistently prognostic for absolute change in percentage of FVC%pred in both the test (p=0·032) and replication (p=0·004) cohorts. Pirfenidone treatment benefit was consistent regardless of baseline biomarker concentration. Interpretation Blood CCL18 concentrations were the most consistent predictor of disease progression across IPF cohorts with potential to inform new target discovery and clinical trial design. Future validation of these findings in prospective studies is warranted. Funding Genentech Inc.

Plasma prorenin is commonly elevated in diabetic patients and appears to predict the development of diabetic nephropathy. However, the pathological role of prorenin is unclear. In the present study, a transgenic, inducible, hepatic prorenin-overexpressing rat model was generated and the effect of prorenin in organ injury was examined. Four groups of rats (cyp1a1 prorenin transgenic male and female rats and non-transgenic littermates) were assigned to receive a diet containing 0.3% of the transgene inducer indole-3-carbinol (I3C) for 4 weeks. Plasma prorenin concentration was increased and mean arterial pressure (MAP) increased from 80 ± 18 to 138 ± 17 (mmHg), whereas renal prorenin/renin protein expression was unchanged, in transgenic rats fed with I3C diet. The intact prorenin, not renin, in plasma and urine samples was further observed by Western blot analysis. Importantly, transgenic rats with high levels of prorenin developed albuminuria, glomerular and tubulointerstitial fibrosis associated with increased expression of transforming growth factor β (TGFβ) 1 (TGFβ1), plasminogen activator inhibitor-1 (PAI-1), collagen, and fibronectin (FN). These rats also exhibited cardiac hypertrophy determined by echocardiography, with elevated ratio of heart weight to body weight (HW/BW). Cardiac collagen in interstitial and perivascular regions was prominent, accompanied by the increase in mRNA contents of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), β-myosin heavy chain (β-MHC), TGFβ1, PAI-1, and collagen in the heart tissue. Furthermore, renal protein levels of p-NF-κB-p65 and monocyte chemoattractant protein-1 (MCP-1), NAPDH oxidases, malondialdehyde (MDA) and 8-isoprostane (8-IP), p-ERK, p-β-catenin, and p-Akt were dramatically increased in prorenin overexpressing rats. These results indicate that prorenin, without being converted into renin, causes hypertension, renal and cardiac fibrosis via the induction of inflammation, oxidative stress and the ERK, β-catenin, and Akt-mediated signals.

T follicular helper (TFH) cells are characteristically defined by their CXCR5 positivity and homing to B cell follicles in secondary lymphoid organs (SLO). An expanded subpopulation of functionally comparable and phenotypically similar PD-1hiCXCR5−CD4+ T cells were recently identified in breast cancer (BC) and rheumatoid arthritis (RA) to have beneficial or detrimental roles, respectively, but are they inflammatory tissue effector TFH cells?

Key findings CHEK1 and circCHEK1_246aa induce multiple myeloma cell proliferation, drug resistance, and bone lesion formation CHEK1 and circCHEK1_246aa evoke myeloma chromosomal instability, partially through CEP170 activation Abstract Multiple myeloma (MM) is characterized by clonal expansion of plasma cells in the bone marrow (BM). Therefore, effective therapeutic interventions must target both myeloma cells and the BM niche. In the present study, we first demonstrated that CHEK1 expression was significantly increased in human MM samples relative to normal plasma cells, and that in MM patients, high CHEK1 expression was associated with poor outcomes. CHEK1 overexpression increased cellular proliferation in MM cells and evoked drug resistance in vitro, while CHEK1 knockdown abrogated this effect. Moreover, CHEK1 was a high-risk gene for poor outcome in MM patients, and, in paired samples from MM patients taken from newly diagnosed and relapsed MM, CHEK1 expression was upregulated. CHEK1-mediated increases in cell proliferation and drug resistance were due in part to CHEK1-induced chromosomal instability (CIN), as demonstrated by Giemsa staining, exon sequencing, and immunofluorescence. CHEK1 activated CIN, partly by phosphorylating CEP170. Interestingly, CHEK1 promoted osteoclast differentiation by direct phosphorylation and activation of NFATc1, indicating that CHEK1 inhibition could target both MM cell proliferation and macrophage osteoclast differentiation in the BM niche. Intriguingly, we also discovered that MM cells expressed circCHEK1_246aa, a circular CHEK1 RNA, which encoded and was translated to the CHEK1 kinase catalytic center. Transfection of circCHEK1_246aa increased MM CIN and osteoclast differentiation similarly to CHEK1 overexpression, suggesting that MM cells could secrete circCHEK1_246aa in the BM niche to increase the invasive potential of MM cells and promote osteoclast differentiation. Finally, we demonstrated in vivo in xenograft models that CHEK1 overexpression prompted MM proliferation and drug resistance, while CHEK1 knockdown conversely inhibited MM growth. Together, these findings suggest that targeting the enzymatic catalytic center encoded by CHEK1 mRNA and circCHEK1_246aa is a promising therapeutic modality to target both MM cells and the BM niche. Figure Disclosures No relevant conflicts of interest to declare.

Although oxide cathodes have been widely used in these Li-ion batteries, these cathodes suffer from instability of the oxygen close-packed structure. In contrast, polyanionic phosphates such as LiFePO4 have incredible lattice stability and safety features owing to the strong covalent bond of P–O, which constrains the oxygen atoms and minimizes the defects of the oxygen site, resulting in stable frameworks. In addition, the presence of the strong P–O covalent bond stabilizes the anti-bonding transitional metal redox couple through an M–O–P inductive effect to generate a relatively high potential. Hence, polyanionic LiFePO4 has been an ideal choice of cathode materials for batteries deployed in electric vehicles. In this review, we revisit the basic features and development of LiFePO4, as an attempt to speeding its future deployment in massive electric vehicles.

// Wang Wang 1, 2, * , Yi Zhang 3, * , Ruini Chen 2, * , Zhidan Tian 4 , Yongpin Zhai 5 , Siegfried Janz 6 , Chunyan Gu 1, 2 and Ye Yang 1, 2 1 The Third Affiliated Hospital, Nanjing University of Chinese Medicine, Nanjing, 210023, China 2 School of Medicine and Life Sciences, Nanjing University of Chinese Medicine, Nanjing, 210023, China 3 Department of Burns and Plastic Surgery, Affiliated Hospital of Nantong University, Nantong, 226001, China 4 Department of Pathology, Nanjing First Hospital, Nanjing, 210006, China 5 Department of Hematology, Jinling Hospital, School of Medicine, Nanjing University, Nanjing, 210002, China 6 Department of Pathology, The University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, 52242, USA * These authors contributed equally to this work Correspondence to: Chunyan Gu, email: guchunyan@njucm.edu.cn Ye Yang, email: yangye876@sina.com Keywords: chromosomal instability, proliferation, drug resistance, multiple myeloma Received: March 09, 2017 Accepted: August 26, 2017 Published: September 11, 2017 ABSTRACT Chromosomal instability (CIN) is an important hallmark of human cancer. CIN not only contributes to all stages of tumor development (initiation, promotion and progression) but also drives, in large measure, the acquisition of drug resistance by cancer cells. Although CIN is a cornerstone of the complex mutational architecture that underlies neoplastic cell development and tumor heterogeneity and has been tightly associated with treatment responses and survival of cancer patients, it may be one of the least understood features of the malignant phenotype in terms of genetic pathways and molecular mechanisms. Here we review new insights into the type of CIN seen in multiple myeloma (MM), a blood cancer of terminally differentiated, immunoglobulin-producing B-lymphocytes called plasma cells that remains incurable in the great majority of cases. We will consider bona fide myeloma CIN genes, methods for measuring CIN in myeloma cells, and novel approaches to CIN-targeted treatments of patients with myeloma. The new findings generate optimism that enhanced understanding of CIN will lead to the design and testing of new therapeutic strategies to overcome drug resistance in MM in the not-so-distant future.

// Chunyan Gu 1, 2, 3, * , Hailin Peng 4, * , Yue Lu 5 , Hongbao Yang 6 , Zhidan Tian 7 , Gang Yin 2 , Wen Zhang 8 , Sicheng Lu 2 , Yi Zhang 9 and Ye Yang 1, 2, 10 1 The Third Affiliated Hospital, Nanjing University of Chinese Medicine, Nanjing 210023, China 2 School of Medicine and Life Science, Nanjing University of Chinese Medicine, Nanjing 210023, China 3 Department of Pathology, School of Medicine, University of Iowa, Iowa City, Iowa 52242, USA 4 Department of Laboratory Medicine, Taizhou People’s Hospital, Taizhou 225300, China 5 Department of Radiation Oncology, Jiangsu Cancer Hospital, Nanjing 210009, China 6 Center for New Drug Safety Evaluation and Research, China Pharmaceutical University, Nanjing 211198, China 7 Department of Pathology, Nanjing First Hospital, Nanjing 210006, China 8 Affiliated Hospital of Stomatology, Nanjing Medical University, Nanjing 210029, China 9 Department of Burns and Plastic Surgery, Affiliated Hospital of Nantong University, Nantong 226001, China 10 Key Laboratory of Acupuncture and Medicine Research of Ministry of Education, Nanjing University of Chinese Medicine, Nanjing 210023, China * These authors have contributed equally to this work Correspondence to: Yi Zhang, email: 198zy@163.com Ye Yang, email: yangye876@sina.com Keywords: multiple myeloma, BTK, senescence, AKT, CGI-1746 Received: February 25, 2017 Accepted: April 04, 2017 Published: May 23, 2017 ABSTRACT We previously explored the role of BTK in maintaining multiple myeloma stem cells (MMSCs) self-renewal and drug-resistance. Here we investigated the elevation of BTK suppressing MM cellular senescence, a state of irreversible cellular growth arrest. We firstly discovered that an increased expression of BTK in MM samples compared to normal controls by immunohistochemistry (IHC), and significant chromosomal gain in primary samples. In addition, BTK high-expressing MM patients are associated with poor outcome in both Total Therapy 2 (TT2) and TT3 cohorts. Knockdown BTK expression by shRNA induced MM cellular senescence using β-galactosidase (SA-b-gal) staining, cell growth arrest by cell cycle staining and decreased clonogenicity while forcing BTK expression in MM cells abrogated these characteristics. We also validated this feature in mouse embryonic fibroblast cells (MEFs), which showed that elevated BTK expression was resistant to MEF senescence after serial cultivation in vitro . Further mechanism study revealed that BTK activated AKT signaling leading to down-regulation of P27 expression and hindered RB activity while AKT inhibitor, LY294002, overcame BTK-overexpression induced cellular senescence resistance. Eventually we demonstrated that BTK inhibitor, CGI-1746, induced MM cellular senescence, colony reduction and tumorigenecity inhibition in vivo . Summarily, we designate a novel mechanism of BTK in mediating MM growth, and BTK inhibitor is of great potential in vivo and in vitro suggesting BTK is a promising therapeutic target for MM.

// Chunyan Gu 1,* , Wen Zhou 2,* , Wang Wang 1 , Hong Xiang 3 , Huiying Xu 2 , Lina Liang 4 , Hua Sui 4 , Libin Zhan 2,3 and Xiaoguang Lu 5 1 School of Medicine and Life Science, Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China 2 Basic Medical College, Nanjing University of Chinese Medicine, Nanjing, Jiangsu, China 3 The Second Affiliated Hospital of Dalian Medical University, Dalian, Liaoning, China 4 Institute of Integrative Medicine, Dalian Medical University, Dalian, Liaoning, China 5 Department of Emergency Medicine, Zhongshan Hospital, Dalian University, Dalian, Liaoning, China * These authors have contributed equally to this work Correspondence to: Libin Zhan, email: // Xiaoguang Lu, email: // Keywords : ZiBuPiYin recipe, diabetes, psychological-stress, cognitive decline, gut microbiota, Pathology Section Received : September 02, 2016 Accepted : January 04, 2017 Published : January 12, 2017 Abstract Numerous researches supported that microbiota can influence behavior and modulate cognitive function through “microbiota-gut-brain” axis. Our previous study has demonstrated that ZiBuPiYin recipe (ZBPYR) possesses excellent pharmacological effects against diabetes-associated cognitive decline. To elucidate the role of ZBPYR in regulating the balance of gut microbiota to improve psychological-stress-induced diabetes-associated cognitive decline (PSDACD), we compared blood glucose, behavioral and cognitive functions and diversity of the bacterial community among experimental groups. The Zucker diabetic fatty (ZDF) rats with PSDACD exhibited behavioral and cognitive anomalies showing as increased anxiety- and depression-like behaviors and decreased learning and memory abilities. High-throughput sequencing of the bacterial 16S rRNA gene revealed that Roseburia and Coprococcus were decreased in ZDF rats with PSDACD compared with control group. Notably, these changes were reversed by ZBPYR treatment. Our findings indicate that ZBPYR might prevent PSDACD by maintaining the compositions of gut microbiota, which could be developed as a new therapy for T2D with PSDACD.

The forkhead box P1 (FOXP1) transcription factor has been shown to regulate the generation and maintenance of quiescent naive murine T cells. In humans, FOXP1 expression has been correlated with overall survival in patients with peripheral T-cell lymphoma (PTCL), although its regulatory role in T-cell function is currently unknown. We found that FOXP1 is normally expressed in all human leukocyte subpopulations. Focusing on primary human CD4+ T cells, we show that nuclear expression of FOXP1 predominates in naive cells with significant downregulation detected in memory cells from blood and tonsils. FOXP1 is repressed following in vitro T-cell activation of naive T cells, and later re-established in memory CD4+ T cells, albeit at lower levels. DNA methylation analysis revealed that epigenetic mechanisms participate in regulating the human FOXP1 gene. ShRNA-mediated FOXP1 repression induces CD4+ T cells to enter the cell cycle, acquire memory-like markers and upregulate helper T-cell differentiation genes. In patients with lymphoproliferative disorders, FOXP1 expression is constitutionally repressed in the clonal T cells in parallel with overexpression of helper T-cell differentiation genes. Collectively, these data identify FOXP1 as an essential transcriptional regulator for primary human CD4+ T cells and suggest its potential important role in the development of PTCL.

Fusarium head blight (FHB), one of the most important diseases of wheat, mainly occurs in the ear. Given that the severity of the disease cannot be accurately identified, the cost of pesticide application increases every year, and the agricultural ecological environment is also polluted. In this study, a neural network (NN) method was proposed based on the red-green-blue (RGB) image to segment wheat ear and disease spot in the field environment, and then to determine the disease grade. Firstly, a segmentation dataset of single wheat ear was constructed to provide a benchmark for the segmentation of the wheat ear. Secondly, a segmentation model of single wheat ear based on the fully convolutional network (FCN) was established to effectively realize the segmentation of the wheat ear in the field environment. An FHB segmentation algorithm was proposed based on a pulse-coupled neural network (PCNN) with K-means clustering of the improved artificial bee colony (IABC) to segment the diseased spot of wheat ear by automatic optimization of PCNN parameters. Finally, the disease grade was calculated using the ratio of the disease spot to the whole wheat ear. The experimental results show that: (1) the accuracy of the segmentation model for single wheat ear constructed in this study is 0.981. The segmentation time is less than 1 s, indicating that the model can quickly and accurately segment wheat ear in the field environment; (2) the segmentation method of the disease spot performed under each evaluation indicator is improved compared with the traditional segmentation methods, and the accuracy is 0.925 in the disease severity identification. These research results can provide important reference value for grading wheat FHB in the field environment, which also can be beneficial for real-time monitoring of other crops’ diseases under near-Earth remote sensing.

Tumor-infiltrating B-cells (TIL-B) in breast cancer (BC) have previously been associated with improved clinical outcomes; however, their role(s) in tumor immunity is not currently well known. This study confirms and extends the correlation between higher TIL-B densities and positive outcomes through an analysis of HER2-positive and triple-negative BC patients from the BIG 02-98 clinical trial (10yr mean follow-up). Fresh tissue analyses identify an increase in TIL-B density in untreated primary BC compared to normal breast tissues, which is associated with global, CD4+ and CD8+ TIL, higher tumor grades, higher proliferation and hormone receptor negativity. All B-cell differentiation stages are detectable but significant increases in memory TIL-B are consistently present. BC with higher infiltrates are specifically characterized by germinal center TIL-B, which in turn are correlated with TFH TIL and antibody-secreting TIL-B principally located in tertiary lymphoid structures. Some TIL-B also interact directly with tumor cells. Functional analyses reveal TIL-B are responsive to BCR stimulation ex vivo, express activation markers and produce cytokines and immunoglobulins despite reduced expression of the antigen-presenting molecules HLA-DR and CD40. Overall, these data support the concept that ongoing humoral immune responses are generated by TIL-B and help to generate effective anti-tumor immunity at the tumor site. ispartof: JCI INSIGHT vol:4 issue:18 ispartof: location:United States status: published

Wheat (Triticum aestivum L.) is one of the three major cereals worldwide. The FusaHum graminearum Sehw., special fugus always damages the wheat ear, and produces vomitoxin,is difficult to control and prevent, and seriously threatens the health of humans, animals and China's food security. Currently, rapidly, accurately and non-destructively diagnostic devices or systems for this disease have not been disclosed. In this study, the infected ears with different severities were picked up in key growth stages. The diseased area of wheat ear was extracted using hypergreen characteristic, and a total of 30 features of infected ears were chosen including color (Lab, HSI, HSV, YCbCr color space), texture (LBP and LLE dimension reduction), and shape (squareness, shape complexity, and eccentricity). Then using the competitive adaptive re-weighted sampling (CARS) and rough set algorithm (RS) to screen the characteristics of the diseased ear, the four characteristics with the largest contribution were determined to establish the CARS-SVM and CARS-RS-SVM models respectively. The study found that the recognition rate of CARS-SVM model is 85.4%, while CARS-RS-SVM model is 92.7%. Thus the CARS-RS-SVM was thought of as the optimal model by two indicators of identification accuracy. On the basis, a wheat scab diagnosis system based on Android mobile phone was constructed. It consists of three parts - Clients, Service-Terminal and Database. The Client was designed by Android Studio and its functions mainly include image acquisition, image storage, GPS positioning, image uploading and diagnostic results display. The Service-Terminal was completed by the mixed programming of Myeclipse and Matlab software, and Tomcat was used as the Server. It mainly implements the functions of image receiving, image preprocessing, feature extraction and selection, and classifier modeling. The MySQL was used to establish two databases: the “Disease Characteristics Database” and the “Disease Diagnosis Knowledge Base”. Finally, through samples testing and validating, the Android-based mobile terminal can real-time collect the image of Fusarium head blight and upload the server. After the target image was processed and compared by the “Disease Characteristics Database”, the appropriate diagnostic knowledge was selected from the “Disease Diagnosis Knowledge Base” and feedbacked to the client. In summary, the results of this study showed that it was helpful for the rapid and non-destructive investigation of infected FHB in the field, and it would provide a reference for the study of other crop diseases, facilitate the application and development of new technologies such as artificial intelligence and big data in agriculture.

Ethnopharmacological relevance Huangkuisiwufang (HKSWF) is composed of Abelmoschus manihot (L.) Medik., Astragalus mongholicus, Polygonum cuspidatum, Curcuma longa L. Abelmoschus Manihot (L.) Medik. has been widely used for the treatment of chronic renal disease, oral ulcers and burn in China for centuries (Committee of the Pharmacopoeia of PR China, 2010). Abelmoschus manihot (L.) Medik., Polygonum cuspidatum, Curcuma longa L. have been mainly applied in folk medicine for their therapeutic effects on diabetes, cancer, heart disease and other diseases. Aim of the study: We aimed to investigate the renoprotective function of HKSWF in anti-Thy nephritis model and clarify the relevant mechanisms. Materials and methods One week after the model of glomerulonephritis created by injecting anti-thymocyte serum (ATS), rats were treated with Huangkui capsule, enalapril or HKSWF by gavage for a period of 8 weeks. The therapeutic effect was evaluated by detection of proteinuria, plasma creatine, blood urea nitrogen (BUN), podocyte injury, glomerular accumulation of extracellular matrix (ECM) and the markers of oxidative stress and renal fibrosis. RNA Sequencing (RNA-seq), KEGG and western blotting analysis were performed to indicate the signaling pathway involved in the therapeutic effect of HKSWF. Results Nephritic rats presented the increase of BUN, serum creatinine (Scr), proteinuria, podocyte damage, glomerular fibrosis, Ang II type 1 receptor (AT1R), and the reduction of creatinine clearance (Ccr). In contrast, application of HKSWF to nephritic rats decreased the levels of BUN and proteinuria, promoted mesangial cell recovery and improved oxidative stress level and podocyte injury. KEGG analysis revealed that pyruvate metabolism was the most significantly upregulated pathway in rats treated with HKSWF compared to disease control group. Increased pyruvate dehydrogenase and PAI-1 caused by nephritis was inhibited by HKSWF interposition. Furthermore, dichloroacetate sodium (DCA), an agonist of pyruvate dehydrogenase, could stimulate PAI-1 expression, which was suppressed by HKSWF. Conclusion Chinese herbal preparation HKSWF has remarkable curative effects on glomerulonephritis animals. HKSWF attenuates pyruvate dehydrogenase to improve glomerular injury.

Cellular adhesion-mediated drug resistance (CAM-DR) occurs frequently in patients with relapsed or refractory multiple myeloma (MM). Elucidating the mechanism underlying CAM-DR and developing the corresponding treatment may prove to be promising for the clinical management of MM. Bruton’s tyrosine kinase (BTK) has been attracting attention in relation to MM progression and drug resistance. BTK was reported to be associated with cell surface CXCR4, a classic cell adhesion molecule and homing factor. However, the exact association between BTK and CAM-DR in MM remains elusive. In this study, we demonstrated that promoting BTK expression induced MM cell adherence to the extracellular matrix (ECM) and stromal cells in vitro and in vivo, and that CAM-DR could be reversed by separating MM cells from ECM or stromal cells. Enhancing BTK expression levels increased CXCR4 expression in MM cells. In addition, BTK may bind directly with CXCR4 and prevent its ubiquitination-induced degradation. Finally, a BTK inhibitor exerted synergistic therapeutic effects with bortezomib in a 5TMM3VT MM mouse model. These findings revealed a novel role of BTK in CAM-DR and may provide a promising approach to MM treatment.

Inducible T cell costimulator (ICOS, cluster of differentiation (CD278)) is an activating costimulatory immune checkpoint expressed on activated T cells. Its ligand, ICOSL is expressed on antigen-presenting cells and somatic cells, including tumour cells in the tumour microenvironment. ICOS and ICOSL expression is linked to the release of soluble factors (cytokines), induced by activation of the immune response. ICOS and ICOSL binding generates various activities among the diversity of T cell subpopulations, including T cell activation and effector functions and when sustained also suppressive activities mediated by regulatory T cells. This dual role in both antitumour and protumour activities makes targeting the ICOS/ICOSL pathway attractive for enhancement of antitumour immune responses. This review summarises the biological background and rationale for targeting ICOS/ICOSL in cancer together with an overview of the principal ongoing clinical trials that are testing it in combination with anti-cytotoxic T lymphocyte antigen-4 and anti-programmed cell death-1 or anti-programmed cell death ligand-1 based immune checkpoint blockade., SCOPUS: re.j, info:eu-repo/semantics/published

Iron is an essential element for biological processes. Iron homeostasis is regulated through several mechanisms, from absorption by enterocytes to recycling by macrophages and storage in hepatocytes. Iron has dual properties, which may facilitate tumor growth or cell death. Cancer cells exhibit an increased dependence on iron compared with normal cells. Macrophages potentially deliver iron to cancer cells, resulting in tumor promotion. Mitochondria utilize cellular iron to synthesize cofactors, including heme and iron sulfur clusters. The latter is composed of essential enzymes involved in DNA synthesis and repair, oxidation‑reduction reactions, and other cellular processes. However, highly increased iron concentrations result in cell death through membrane lipid peroxidation, termed ferroptosis. Ferroptosis, an emerging pathway for cancer treatment, is similar to pyroptosis, apoptosis and necroptosis. In the present review, previous studies on the physiology of iron metabolism and its role in cancer are summarized. Additionally, the significance of iron regulation, and the association between iron homeostasis and carcinogenic mechanisms are discussed.

Age-related genetic changes in lymphocyte subsets are not currently well documented. BACH2 is a transcription factor that plays an important role in immune-mediated homeostasis by tightly regulating PRDM1 expression in both B-cells and T-cells. BACH2 gene expression is highly sensitive to DNA damage in aged mice. This concept led us to investigate the variation in BACH2 and also PRDM1 expression in major lymphocyte subsets with age., info:eu-repo/semantics/published

Background Following up on previous work demonstrating the involvement of the transcription factor forkhead box M1 (FOXM1) in the biology and outcome of a high-risk subset of newly diagnosed multiple myeloma (nMM), this study evaluated whether FOXM1 gene expression may be further upregulated upon tumor recurrence in patients with relapsed multiple myeloma (rMM). Also assessed was the hypothesis that increased levels of FOXM1 diminish the sensitivity of myeloma cells to commonly used myeloma drugs, such as the proteasome inhibitor bortezomib (Bz) and the DNA intercalator doxorubicin (Dox). Methods FOXM1 message was evaluated in 88 paired myeloma samples from patients with nMM and rMM, using gene expression microarrays as measurement tool. Sources of differential gene expression were identified and outlier analyses were performed using statistical methods. Two independent human myeloma cell lines (HMCLs) containing normal levels of FOXM1 (FOXM1N) or elevated levels of lentivirus-encoded FOXM1 (FOXM1Hi) were employed to determine FOXM1-dependent changes in cell proliferation, survival, efflux-pump activity, and drug sensitivity. Levels of retinoblastoma (Rb) protein were determined with the assistance of Western blotting. Results Upregulation of FOXM1 occurred in 61 of 88 (69%) patients with rMM, including 4 patients that exhibited > 20-fold elevated expression peaks. Increased FOXM1 levels in FOXM1Hi myeloma cells caused partial resistance to Bz (1.9–5.6 fold) and Dox (1.5–2.9 fold) in vitro, using FOXM1N myeloma as control. Reduced sensitivity of FOXM1Hi cells to Bz was confirmed in vivo using myeloma-in-mouse xenografts. FOXM1-dependent regulation of total and phosphorylated Rb agreed with a working model of myeloma suggesting that FOXM1 governs both chromosomal instability (CIN) and E2F-dependent proliferation, using a mechanism that involves interaction with NIMA related kinase 2 (NEK2) and cyclin dependent kinase 6 (CDK6), respectively. Conclusions These findings enhanced our understanding of the emerging FOXM1 genetic network in myeloma and provided preclinical support for the therapeutic targeting of the FOXM1-NEK2 and CDK4/6-Rb-E2F pathways using small-drug CDK and NEK2 inhibitors. Clinical research is warranted to assess whether this approach may overcome drug resistance in FOXM1Hi myeloma and, thereby, improve the outcome of patients in which the transcription factor is expressed at high levels. Electronic supplementary material The online version of this article (10.1186/s12885-018-5015-0) contains supplementary material, which is available to authorized users.

Objective Fatal central nervous system (CNS) malformation is one of the most common congenital malformations, and is an important cause of infant mortality. Ultrasound diagnostic technology has the advantages of fast, safe, economic, convenient and real-time dynamic imaging, and it is the first choice for imaging diagnosis of the foetus. This paper studies the diagnosis of fatal central nervous system malformations based on prenatal Doppler ultrasound, and further accumulates clinical experience for prenatal diagnosis. Methods 320 pregnant women who underwent prenatal diagnosis in our hospital from August 2017 to December 2018. Ultrasound showed abnormal fatal craniocerebral development and MRI examination of fatal cranial brain was performed on the same day or the next day. The PHILIPS IU22 colour Doppler ultrasound system was used to scan pregnant women. Ultrasound showed 320 cases of fatal craniocerebral abnormalities and MRI diagnosis on the same day or the next day, of which 122 cases were consistent with MRI results, accounting for 38%; 173 cases were supplemented with MRI diagnostic information, accounting for 54%; MRI corrected diagnostic information There were 16 cases, accounting for 5%; 9 cases were abnormal, but 3 cases were not revealed by MRI. Conclusion Ultrasound examination during pregnancy has a high detection rate for the diagnosis of fatal CNS malformations. As the preferred imaging method for prenatal diagnosis, it is essential for timely and early screening of fatal malformations. MRI has a higher diagnostic value for fatal CNS malformations, and can play an important role in supplementing and misdiagnosing cases that have been missed by ultrasound. Therefore, in the prenatal diagnosis, attention should be paid to combining these two inspection methods, each taking its advantages and applying it to clinical.

As a result of an author oversight in the original article [1], the legend of Figure 5A and C is inaccurate and one panel in Figure 5C (FOXM1N H929 cells shown in the top row, left) is wrong.

Tertiary lymphoid structures (TLS) have been detected in several types of human solid tumors. These structures are thought to regulate local adaptive immune responses that can promote or antagonize tumor progression. Despite positive prognostic values associated with a TLS presence in several studies, discrepancies still exist. TLS are structurally organized entities composed of varying numbers of multiple cell types making their assessment in tumor tissues, particularly biopsies, challenging. Immunohistochemical staining of TLS-related cell populations is the most frequently used method for identifying and scoring them; however, TLS-related gene expression has also been explored. The protocols described are detailed to allow the user to quantify TLS-related gene expression on formalin-fixed paraffin-embedded human breast tumor tissues.

Purpose: In recent years, with the emergence of targeted proteasome inhibitors (PIs), the treatment of multiple myeloma (MM) has made great progress and significantly improves the survival rate of patients. However, MM remains an incurable disease, mainly due to the recurrence of drug resistance. The constitutive photomorphogenic 1 (RFWD2, also known as COP1), is closely related to the occurrence and development of tumors, but its role in MM is largely unknown. This study was aimed to explore the mechanism of RFWD2 on cell proliferation and resistance to proteasome inhibitor in MM. Experimental Design: Using gene expression profiling (GEP) samples, we verified the relation of RFWD2 to MM patients' survival and drug-resistance. The effect of RFWD2 on cell proliferation was confirmed by MTT and cell cycle analysis in RFWD2-overexpressed and RFWD2-knockdown MM cells. MTT and apoptosis experiments were performed to evaluate whether RFWD2 influenced the sensitivity of MM cells to several chemotherapy drugs. MM xenografts were established in immunodeficient NOD/SCID mice by injecting wild-type or RFWD2 over-expression MM cells with drug intervention. The mechanism of drug resistance was elucidated by analyzing the association of RFWD2 with E3 ligase of p27. Bortezomib-resistant RPMI 8226 cells were used to construct RFWD2 knockdown cells, which were injected into NOD/SCID mice to assess the effect of RFWD2 on bortezomib resistance in vivo. Results: RFWD2 expression was closely related to poor outcome, relapse and bortezomib resistance in MM patients' GEP cohorts. Elevated RFWD2 induced cell proliferation, while decreased RFWD2 inhibited cell proliferation and induced apoptosis in MM cells. RFWD2-overexpression MM cells resulted in PIs resistance, however, no chemotherapy resistance to adriamycin and dexamethasone was observed in vitro. In addition, overexpressing RFWD2 in MM cells led to bortezomib resistance rather than adriamycin resistance in myeloma xenograft mouse model. RFWD2 regulated the ubiquitination degradation of P27 by interacting with RCHY1 ubiquitin ligase. The knockdown of RFWD2 in bortezomib-resistant RPMI 8226 cells overcame bortezomib resistance in vivo. Conclusions: Our data demonstrate that elevated RFWD2 induces MM cell proliferation and resistance to PIs, but not to adriamycin and dexamethasone both in vitro and in vivo through mediating the ubiquitination of p27. Collectively, RFWD2 is a novel promising therapeutic target in MM. Disclosures No relevant conflicts of interest to declare.

Proteasome inhibition demonstrates highly effective impact on multiple myeloma (MM) treatment. In this study we aimed to examine a novel well tolerated orally applicable proteasome inhibitor NNU546 and its hydrolyzed pharmacologically active form NNU219. NNU219 showed more selective inhibition to proteasome catalytic subunits and less off-target effect than bortezomib ex vivo. Furthermore, intravenous and oral administration of NNU219 and NNU546, respectively, led to a more sustained pharmacodynamic inhibition of proteasome activities compared with bortezomib. Intriguingly, NNU219 exhibited potential anti-MM activity on both MM cell lines and primary samples in vitro. The anti-MM activity of NNU219 was determined to be associated with induction of G2/M-phase arrest, as well as induction of apoptosis via activation of the caspase cascade and endoplasmic reticulum stress response. At well-tolerated doses, significant growth-inhibitory effects of NNU219 and NNU546 were observed in 3 different human MM xenograft mouse models and significant anti-MM activity was observed even in the presence of a bone marrow microenvironment. Taken together, these findings provided the basis for clinical trials of NNU546 to determine its potential as a candidate for MM treatment. Disclosures No relevant conflicts of interest to declare.

In order to identify immunodominant linear B cell epitopes in the nucleocapsid protein N of severe fever with thrombocytopenia syndrome virus(SFTSV),bioinformatics programs were used to analyze antigenicity, hydrophilicity and surface probability of the amino acid sequence and predict possible linear B cell epitopes. PyMOL software was used to analyze the distribution of linear B cell epitopes in nucleocapsid protein N based on its crystal structure. Corresponding peptides were synthesized and examined in peptide enzyme-linked immunosorbent assay(Peptide-ELISA)individually to check whether they reacted with sera from SFTSV-infected patients. As a result, a total of six potential linear B cell epitopes were predicted as the following: A(40-KKLKETGGDDWVKDTK-55), B(71-ASGKMSNSGSKRL-83), C(94-ERAETRL-100),D(135-LKVENYPP-142),E(157-GVSEATT-163)and F(184-KMRGASKTEVYNSFRDP-200).All epitopes were located on the surface of the nucleocapsid protein N and contained flexible loops. Each of the six synthetic peptides reacted positively with sera from SFTSV-infected patients and were identified as immunodominant linear B cell epitopes. Linear regression analysis showed a positive correlation between each peptide-ELISA and commercialized N protein-based EIA. In this study, immunodominant linear B cell epitopes from the nucleocapsid protein N of SFTSV were successfully predicted and confirmed. These findings may help to establish the molecule basis of specific antigenicity and disease diagnosis.

Cytotoxic CD4 (CD4CTX) T cells are emerging as an important component of anti-viral and anti-tumor immunity but the molecular basis of their development remains poorly understood. In the context of human cytomegalovirus infection, a significant proportion of CD4 T cells displays cytotoxic functions. We observed that the transcriptional program of these cells was enriched in CD8 T cell lineage genes despite the absence of ThPOK downregulation. We further show that establishment of CD4CTX-specific transcriptional and epigenetic programs occurred in a stepwise fashion along the Th1-differentiation pathway. In vitro, prolonged activation of naive CD4 T cells in presence of Th1 polarizing cytokines led to the acquisition of perforin-dependent cytotoxic activity. This process was dependent on the Th1 transcription factor Runx3 and was limited by the sustained expression of ThPOK. This work elucidates the molecular program of human CD4CTXT cells and identifies potential targets for immunotherapy against viral infections and cancer., SCOPUS: ar.j, info:eu-repo/semantics/published

Tertiary lymphoid structures (TLS) have been detected in several types of human solid tumors. These structures are thought to regulate local adaptive immune responses that can promote or antagonize tumor progression. Despite positive prognostic values associated with a TLS presence in several studies, discrepancies still exist. TLS are structurally organized entities composed of varying numbers of multiple cell types making their assessment in tumor tissues, particularly biopsies, challenging. Immunohistochemical staining of TLS-related cell populations is the most frequently used method for identifying and scoring them; however, TLS-related gene expression has also been explored. The protocols described are detailed to allow the user to quantify TLS-related gene expression on formalin-fixed paraffin-embedded human breast tumor tissues.

Mounting evidence has shown that microRNAs (miRNAs), a class of small non-coding RNAs, are frequently deregulated in human malignancies and have pivotal roles in diverse biological processes including cancer cell proliferation. Herein, we investigated the expression pattern of miR-383 in 64 hepatocellular carcinoma (HCC) tissues and 4 HCC cell lines and found that miR-383 was downregulated in HCC tissues and cell lines. Moreover, miR-383 expression in HCC was significantly correlated with tumor size and tumor-node-metastasis (TNM) stage. Kaplan-Meier analysis showed that decreased miR-383 expression was associated with poor overall survival of HCC patients. In addition, Cox regression analysis indicated that miR-383 was an independent prognostic factor for HCC patients. Then, functional studies demonstrated that ectopic miR-383 expression could significantly suppress the in vitro proliferation of HCC cells, as well as induce cell cycle arrest and cell apoptosis. Luciferase reporter assay further identified that a proliferation-inducing ligand (APRIL), a member in the tumor necrosis factor (TNF) superfamily, was a novel target gene for miR-383. Subsequent investigation revealed that miR-383 expression was inversely correlated with APRIL messenger RNA (mRNA) expression in HCC tissues. Besides, recombinant human APRIL (rhAPRIL) could rescue HCC cell proliferation inhibited by miR-383. Taken together, our present study provided the first evidence that miR-383 was decreased in HCC and associated with tumor progression and prognosis of HCC patients. Furthermore, our findings confirmed that miR-383 might inhibit HCC cell proliferation partially via downregulating APRIL expression. Thus, this study might provide a promising strategy by targeting with the miR-383-APRIL axis in the treatment of HCC.

Summary Understanding the regulation of islet cell mass has important implications for the discovery of regenerative therapies for diabetes. The liver plays a central role in metabolism and the regulation of endocrine cell number, but liver-derived factors that regulate α-cell and β-cell mass remain unidentified. We propose a nutrient-sensing circuit between liver and pancreas in which glucagon-dependent control of hepatic amino acid metabolism regulates α-cell mass. We found that glucagon receptor inhibition reduced hepatic amino acid catabolism, increased serum amino acids, and induced α-cell proliferation in an mTOR-dependent manner. In addition, mTOR inhibition blocked amino-acid-dependent α-cell replication ex vivo and enabled conversion of α-cells into β-like cells in vivo. Serum amino acids and α-cell proliferation were increased in neonatal mice but fell throughout postnatal development in a glucagon-dependent manner. These data reveal that amino acids act as sensors of glucagon signaling and can function as growth factors that increase α-cell proliferation.

The current study examines the relationships of several psychosocial correlates and mediating factors with problem behaviors among children (6–18 years of age) affected by parental HIV/AIDS in rural China. Cross-sectional data were collected from 755 children orphaned by AIDS (“orphans”) and 466 children living with HIV-positive parents (“vulnerable children”) during 2006–2007 in central rural China. The structural equation modeling analyses revealed that HIV-related stigma had a positive direct effect on problem behaviors of vulnerable children, while HIV-related stigma and low education aspiration had direct negative effects on school adjustment among both orphans and vulnerable children. Measures of future orientation and perceived social support mediated the effect of HIV-related stigma and low education aspiration on school adjustment, which in turn was negatively associated with problem behaviors of both groups. The data also showed that school adjustment mediated the effect of low education aspiration and HIV-related stigma on problem behaviors of both orphans and vulnerable children. Future prevention intervention efforts aiming to reduce problem behaviors and improve school adjustment among these children should target multiple factors at individual, community, and societal levels such as enhancing the children’s future expectation, increasing the social support from family, school, and community, and reducing HIV-related stigma against children and families affected by HIV.