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Tumor buds are usually defined as isolated single cancer cells or clusters of up to four cancer cells located at the front of invasive tumors which play an important role in the clinical pathological study of colorectal cancer. The prognostic value of tumor budding in colorectal cancer has been supported by a large amount of evidence. However, its molecular mechanism remains unclear, and it is also a research hotspot now. This paper reviews the latest research progress on the molecular mechanisms of tumor budding in colorectal cancer.

Objectives This study aimed to assess the health utility of leukemia patients in China using the EQ-5D-5L, compare it with the population norms, and identify the potential factors associated with health utility. Methods A hospital based cross-sectional survey was conducted in three tertiary hospitals from July 2015 to February 2016. A total of 186 patients with leukemia completed the EQ-5D-5L and their health utility scores were calculated using the Chinese value set. EQ-5D-5L utility and dimensions scores of leukemia patients were compared with China’s population norms using Kruskal–Wallis test and chi square test. Potential factors associated with health utility were identified using Tobit regression. Results The mean EQ-5D-5L utility scores of patients with leukemia, grouped by either gender or age, were significantly lower than those of the general population (p p Conclusion This study indicated that leukemia patients have worse health status compared to the general population of China and that multiple factors affect the health utility of the patients. The utility scores reported in this study could be useful in future cost-utility analysis.

Polymorphisms in arsenic (+ 3 oxidation state) methyltransferase (AS3MT) have been shown to be related to interindividual variations in arsenic metabolism and to influence adverse health effects in acute promyelocytic leukemia (APL) patients treated with arsenic trioxide (As2O3). The occurrence of hyperleukocytosis with As2O3 treatment seriously affects the early survival rate of APL patients, but no definite explanation for such a complication has been clearly established. To clarify the causes of this situation, AS3MT polymorphisms 14215 (rs3740390), 14458 (rs11191439), 27215 (rs11191446), and 35991 (rs10748835) and profiles of plasma arsenic metabolites were evaluated in a group of 54 newly diagnosed APL patients treated with single-agent As2O3. High-performance liquid chromatography–hydride generation-atomic fluorescence spectrometry (HPLC–HG-AFS) was used to determine the concentrations of plasma arsenic metabolites. Plasma arsenic methylation metabolism capacity was evaluated by the percentage of inorganic arsenic (iAs), monomethylarsonic acid (MMA), dimethylarsinic acid (DMA), primary methylation index (PMI, MMA/iAs), and secondary methylation index (SMI, DMA/MMA). The results showed that APL patients who developed hyperleukocytosis had a higher plasma iAs%, but a lower MMA% and PMI than those who did not develop hyperleukocytosis during As2O3 treatment. In addition, patients with the AS3MT 14215 (rs3740390) CC genotype had significantly higher plasma iAs% and incidence of hyperleukocytosis, but lower PMI than patients with the CT + TT genotype. Conversely, we did not observe statistically significant associations between the occurrence of hyperleukocytosis and AS3MT 14458 (rs11191439), 27215 (rs11191446), and 35991 (rs10748835) polymorphisms in our study subjects. These results indicated that AS3MT 14215 (rs3740390) might be used as an indicator for predicting the occurrence of hyperleukocytosis in APL patients treated with As2O3.

Polo-like kinase 1 (PLK-1) is a protein kinase that plays a significant role in the initiation, maintenance, and completion of mitotic processes in the cell cycle. PLK-1 has been recorded to be over-expressed in various human cancers and is associated with poor prediction; thus it is an attractive target for anticancer therapy. Novel α-santalol functionalized chitosan nanoparticles were synthesized using the sol gel method and were assessed for their in vitro (MTT, apoptotic staining assays, and cell cycle analysis) and in vivo activities. α-Santalol loaded chitosan NPs inhibited the proliferation of triple negative breast cancer (MDA-MB-231) at an inhibitory concentration of (IC50) about 4.5 μg mL; meanwhile, in normal cells, no adverse effects were exhibited up to 100 μg mL−1. The findings also implicated a decreased expression of the anti-apoptotic protein, BCL-2 with PLK-1 and an increase in the expression of BAD, caspases and BAX. However, in in vivo studies, the treated animal group exhibited no aberrant effects in vital organs or blood parameters. Tumor growth was significantly inhibited after i.v. injection of α-santalol loaded chitosan NPs at a dose of 5 mg kg−1. Taken together, the α-santalol functionalized chitosan NPs hold great potential in biomedical applications, especially cancer theranostics, due to their versatile nature as well as diagnostics for clinical tumor biology.

Annexin A2 (ANXA2) is reported to be associated with cancer development. To investigate the roles ANXA2 plays during the development of cancer, the RNAi method was used to inhibit the ANXA2 expression in caco2 (human colorectal cancer cell line) and SMMC7721 (human hepatocarcinoma cell line) cells. The results showed that when the expression of ANXA2 was efficiently inhibited, the growth and motility of both cell lines were significantly decreased, and the development of the motility relevant microstructures, such as pseudopodia, filopodia, and the polymerization of microfilaments and microtubules were obviously inhibited. The cancer cell apoptosis was enhanced without obvious significance. The possible regulating pathway in the process was also predicted and discussed. Our results suggested that ANXA2 plays important roles in maintaining the malignancy of colorectal and hepatic cancer by enhancing the cell proliferation, motility, and development of the motility associated microstructures of cancer cells based on a possible complicated signal pathway.

Human herpes virus 6 (HHV-6) is a ubiquitous and most common pathogen that affects humans. Human herpes virus 6B (HHV-6B) is a wide spread human herpesvirus that infects most people when they are children, establishes latent infections in the central nervous system (CNS), especially in the hippocampus and amygdala, and induces neurologic diseases. HHV-6 can establish a latent infection and be reactivated by various stimuli. Recently, viral genomic DNA of HHV-6B has been detected in surgically removed brain tissues of intractable epilepsy patients, suggesting the involvement of HHV-6B in the pathogenesis of epilepsy. Temporal lobe epilepsy (TLE) has been shown to be closely related with HHV-6B. TLE patients with HHV-6B in their brains suffer from reiterative attacks of febrile seizures and hippocampal sclerosis. However, the mechanisms underlying the contribution of this virus to the development of TLE remains unknown. The direct damage and immune activation caused by the virus are involved in the process of neuron damage, abnormal neural circuit formation and glial cell proliferation. In addition, some cytokines like interleukin-17A (IL-17A), nuclear factor-kappa B (NF-κb), transforming growth factor-β (TGF-β), mitogen-activated protein kinase (MAPK) and phospholipase A2 are up-regulated and involved in the pathological process of TLE. More studies are needed to clarify the mechanisms underlying the link between HHV-6B and epilepsy, and identify biomarkers to recognize different patient groups for anti-inflammatory or immunomodulatory therapies.

Objective: To investigate the expression and clinical significance of hypoxia inducible factors HIF-1a, Gli-1 and MMP9 in breast cancer. Methods: Eighty patients with invasive ductal carcinoma of the breast and 40 normal tissues adjacent to cancer were selected. Immunohistochemical methods were used to detect the expression of HIF-1a, Gli-1 and MMP9 in breast cancer and normal tissues adjacent to cancer, and their relationship with clinicopathological features of breast cancer and prognosis was explored. Results: The positive rates of HIF-1a, Gli-1 and MMP9 in breast cancer tissues were significantly higher than those in normal breast tissues. HIF-1a, Gli-1 and MMP9 expressions are positively correlated in breast cancer. Conclusion: HIF-1a, Gli-1 and MMP9 proteins are involved in the pathogenesis of breast cancer.

Ethnopharmacological relevance The traditional Chinese formula Danning tablets exhibit wide clinical applications in liver and gallbladder diseases, and currently it is reported to be effective on fatty liver disease in clinical trials. However, the underlying mechanisms remain elusive. Aim of the study The purpose of the present study was to assess the effects and potential pharmacological mechanisms of Danning tablet against high fat diet (HFD)-induced obesity, fatty liver, and related metabolic disorders in mice. Materials and methods C57BL/6 J male mice were treated with HFD for 12 weeks to trigger obesity and fatty liver condition. Then those mice were randomly divided into 5 groups, namely HFD, Danning tablet (0.75, 1.5 or 3 g/kg bodyweight) or lovastatin (30 mg/kg bodyweight) for extra 6 weeks’ treatment of HFD. Food intake and bodyweight were recorded each week. In the last week, before the mice were sacrificed, fasting blood glucose levels and insulin levels were measured. Furthermore, insulin and glucose tolerance tests were performed. Blood and hepatic lipid levels were examined, the lipid metabolism-associated gene expressions and protein levels in the liver or adipose tissues were assayed after sacrificing all mice. Results Our results demonstrated that a high dose of Danning tablet (3 g/kg) treatment mitigated body weight gain, reduced blood and hepatic cholesterol and triglyceride levels. The morphology analysis showed that Danning tablets could reduce lipid accumulation in both liver and brown adipose tissue. Moreover, Danning tablets could improve fasting blood glucose levels and ameliorate glucose and insulin tolerance in HFD-induced obese mice. Furthermore, qRT-PCR analysis revealed that the mRNA expressions of SREBP-1 and SREBP-2 as well as their target genes were remarkedly down-regulated in the liver and adipose tissue of diet-induced obesity (DIO) mice after treating those mice with Danning tablets. Conclusion Our results indicated that Danning tablets could improve the obesity-induced metabolic associated fatty liver disease (MAFLD) and related metabolic disorders. The potential mechanism may probably involve the regulation of the SREBP pathway.

A 20-year-old female presented with fine motor deficits and visual field defect was admitted to our hospital. CSF tests for autoimmune encephalitis antibodies and onconeuronal antibodies were unremarkable. MRI showed unilateral lesion involving left basal ganglia, external capsule, insula, hippocampus, and amygdala, which was considered to be demyelinating pseudotumor after surgical intervention. The patient's symptoms relieved so she didn't consent to immunotherapy. Two years and a half later our patient reported sudden anosmia. Reexamination by MRI demonstrated a new lesion. We then detected anti-CASPR2 antibodies in the patient's serum and gave her immunotherapy.

Objective: To explore the expression of human myxovirus resistance protein A (MxA) in triple negative breast cancer (TNBC) and its relationship with prognosis. Methods: 144 cases of TNBC confirmed by pathology before or after surgery from January 2014 to January 2017 in the First Central Hospital of Baoding City were retrospectively collected. Western blotting was used to detect the expression of MxA protein in TNBC and adjacent breast tissues. According to the expression of MxA protein, 144 TNBC patients were divided into low MxA protein expression group (n = 91) and MxA protein high expression group (n = 53) for subsequent comparison of prognosis of patients in between these two groups. Results: The expression of MxA protein in TNBC tissue was lower than that of adjacent breast tissue, and the difference was statistically significant (P

Nitrogen-containing bisphosphonates (N-BPs), such as alendronate, are the most widely prescribed medications for diseases involving bone, with nearly 200 million prescriptions written annually. Recently, widespread use of N-BPs has been challenged due to the risk of rare but traumatic side effects such as atypical femoral fracture (AFFs) and osteonecrosis of the jaw (ONJ). N-BPs bind to and inhibit farnesyl diphosphate synthase (FDPS), resulting in defects in protein prenylation. Yet it remains poorly understood what other cellular factors might allow N-BPs to exert their pharmacological effects. Here, we performed genome-wide studies in cells and patients to identify the poorly characterized gene, ATRAID. Loss of ATRAID function results in selective resistance to N-BP-mediated loss of cell viability and the prevention of alendronate-mediated inhibition of prenylation. ATRAID is required for alendronate inhibition of osteoclast function, and ATRAID-deficient mice have impaired therapeutic responses to alendronate in both postmenopausal and senile (old age) osteoporosis models. Lastly, we performed exome sequencing on patients taking N-BPs that suffered ONJ or an AFF. ATRAID is one of three genes that contain rare non-synonymous coding variants in patients with ONJ or AFF that is also differentially expressed in poor outcome groups of patients treated with N-BPs. We functionally validated this patient variation in ATRAID as conferring cellular hypersensitivity to N-BPs. Our work adds key insight into the mechanistic action of N-BPs and the processes that might underlie differential responsiveness to N-BPs in people.One Sentence SummaryATRAID is essential for responses to the commonly prescribed osteoporosis drugs nitrogen-containing bisphosphonates.OverlineBONE

This study aimed to evaluate treatment response, survival, safety profiles, and predictive factors to chimeric antigen receptor T cell (CAR-T) therapy in Chinese patients with relapsed or refractory B cell acute lymphoblast leukemia (R/R B-ALL). 39R/R B-ALL patients who underwent CAR-T therapy were included. Baseline data were collected from patients’ electronic medical records. Patients’ peripheral bloods, bone marrow aspirates, and biopsies were obtained for routine examination, and treatment response and survival profiles as well as adverse events were evaluated. The rates of complete remission (CR), CR with minimal residual disease (MRD) negative/positive, and bridging to hematopoietic stem-cell transplantation (HSCT) were 92.3%, 76.9%, 15.4%, and 43.6%, respectively. The median event-free survival (EFS) was 11.6 months (95% confidence interval (CI): 4.0–19.2 months) and median overall survival (OS) was 14.0 months (95% CI: 10.9–17.1 months). Bridging to HSCT independently predicted better EFS and OS, while high bone marrow blasts level independently predicted worse EFS. The incidence of cytokine release syndrome (CRS) was 97.4%, and refractory disease as well as decreased white blood cell independently predicted higher risk of severe CRS. Other common adverse events included hematologic toxicities (grade I: 5.1%, grade II: 7.7%, grade III: 17.9%, grade IV: 69.2%), neurotoxicity (28.2%), infection (38.5%), and admission for intensive care unit (10.3%). In conclusion, CAR-T therapy presents with promising treatment response, survival and safety profiles, and higher disease burden predicts worse survival as well as increased risk of severe CRS in Chinese R/R B-ALL patients.

Objective In recent years, research on microRNAs (miRNAs) associated with coronary artery disease (CAD) has attracted considerable attention. However, findings of these studies on the validity of circulating miRNAs in CAD diagnosis are controversial. A meta-analysis was therefore conducted to determine the potential value of miRNAs as biomarkers in CAD diagnosis. Methods Relevant documents on miRNAs expression levels in the diagnosis of CAD were searched and collected from Pubmed, Embase, and Web of Science. They were collected from the time of inception of the database till January 31, 2020. A meta-analysis was conducted using Stata14.0 software. Forest maps were studied and a comprehensive evaluation of the diagnostic value of the expression levels of mRNAs in CAD was conducted using statistical indicators such as the summary receiver operating characteristic curve. Results Overall, 14 studies were included, with 38 data sets, involving 29 miRNAs with 846 cases and 898 controls. The meta-analysis revealed that the average sensitivity and specificity of miRNAs for CAD diagnosis were 0.80 (0.75-0.84) and 0.78 (0.75-0.81), respectively. The positive likelihood, negative likelihood, and diagnostic odds ratios were 3.7 (3.1-4.4), 0.26 (0.21-0.33), and 14 (10-21), respectively, and the area under the curve was 0.85 (0.82-0.88). Subgroup analysis revealed that the accuracy in the Asian population was higher than that in the non-Asian population. Multiple miRNAs may be more diagnostically accurate than single miRNAs. MiRNAs in whole blood were more accurate than those in plasma, serum, and peripheral blood mononuclear cells. The diagnostic performance of the quantitative real-time polymerase chain reaction group was better than that of the qPCR group. Conclusion According to our study, miRNAs may be a new, non-invasive diagnostic tool for the diagnosis of CAD. As a screening tool in clinical practice, it has potential diagnostic value and is worthy of clinical promotion. Considering the number and quality of the studies included in this meta-analysis, the above conclusion requires more quality research to verify it.

Background Non-small cell lung cancer (NSCLC) chemoresistance usually limits the clinical efficacy of chemotherapeutic approaches. However, few reports have revealed the regulation of miR-135b and Frizzled-1 (FZD1) involved in NSCLC chemoresistance. Methods To identify the mechanism of miR-135b and FZD1 in NSCLC chemoresistance and to observe their biological functions, we detected the expression levels of miR-135b and FZD1 by conducting quantitative real-time polymerase chain reaction (RT-qPCR) and modified the expressions of miR-135b and FZD1 by transiently transfecting cells with miR-135b mimics or FZD1-siRNA. The 3′-untranslated region (3′-UTR) of FZD1 combined with miR-135b was verified through dual-luciferase reporter assay. Results Compared with that in A549 parental cell lines, the miR-135b expression in drug-resistant lung cancer cell lines (A549/DDP) was decreased and their FZD1 expression was increased. The increased miR-135b expression and silenced FZD1 expression enhanced the sensitivity of resistant cells to cisplatin treatment. The high expression of miR-135b in A549/DDP cells remarkably decreased the mRNA levels of FZD1. FZD1 was further identified as the functional downstream target of miR-135b by directly targeting the 3′-UTR of FZD1. Conclusion The amplification of miR-135b suppressed NSCLC chemoresistance by directly mediating the FZD1 downregulation.

miR-133a has been demonstrated to play an important role in tumor progression. The aim of present study was to analyze the correlation between miR-133a expression level and clinicopathologic features and its prognostic significance in non-small cell lung cancer (NSCLC). The expression of miR-133a in 104 pairs of human lung cancer tissues and adjacent normal lung tissues were analyzed by qRT-PCR. Here we show that miR-133a was down-regulated in NSCLC. The levels of miR-133a were negatively correlated with the status of N classification (N0-N1 vs. N2-N3, P=0.000), clinical stage (I-II vs. III-IV, P=0.010) and MMP-14 expression (High vs. Low, P=0.012). The patients with low miR-133a expression had shorter survival time than those with high miR-133a expression. Multivariate analysis indicated that the level of miR-133a expression was an independent prognostic indicator (P=0.012) for the survival of patients with NSCLC. In conclusion, decreased expression of miR-133a might be a potential unfavorable prognostic factor for patients with NSCLC, and further studies would be needed to prove our findings.

// Yingmei Zhang 1, * , Jinxiao Hou 2, * , Fei Ge 2 , Fenglin Cao 1 , Haitao Li 2 , Ping Wang 2, 3 , Mengyuan Xu 2 , Peng Song 1 , Xiaoxia Li 2 , Shuye Wang 2 , Jinmei Li 2 , Xueying Han 2 , Yanhong Zhao 2 , Yanhua Su 2 , Yinghua Li 2 , Shengjin Fan 2 , Limin Li 2 , Jin Zhou 1, 2 1 Central Laboratory, The First Affiliated Hospital, Harbin Medical University, Harbin, China 2 Department of Hematology, The First Affiliated Hospital, Harbin Medical University, Harbin, China 3 Department of Neonatology, The First Affiliated Hospital, Harbin Medical University, Harbin, China * These authors have contributed equally to this work Correspondence to: Jin Zhou, email: jinzhou1111@126.com Keywords: acute promyelocytic leukemia, differentiation syndrome, microRNA, mRNA, microarrays Received: January 08, 2016 Accepted: September 02, 2016 Published: September 13, 2016 ABSTRACT The pathogenesis of therapy-induced differentiation syndrome (DS) in patients with acute promyelocytic leukemia (APL) remains unclear. In this study, mRNA and microRNA (miRNA) expression profiling of peripheral blood APL cells from patients complicated with vs. without DS were integratively analyzed to explore the mechanisms underlying arsenic trioxide treatment-associated DS. By integrating the differentially expressed data with the data of differentially expressed microRNAs and their computationally predicted target genes, as well as the data of transcription factors and differentially expressed target microRNAs obtained from a literature search, a DS-related genetic regulatory network was constructed. Then using an EAGLE algorithm in clusterViz, the network was subdivided into 10 modules. Using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database the modules were annotated functionally, and three functionally active modules were recognized. The further in-depth analyses on the annotated functions of the three modules and the expression and roles of the related genes revealed that proliferation, differentiation, apoptosis and infiltration capability of APL cells might play important roles in the DS pathogenesis. The results could improve our understanding of DS pathogenesis from a more overall perspective, and could provide new clues for future research.

Many patients with psoriasis have developed acute promyelocytic leukemia (APL) whereas few reports on psoriasis-associated APL were found in the published literature. This study was aimed to study the etiology, clinical characteristics, and prognosis of psoriasis-associated APL and to map a suitable treatment regime for this condition.This study retrospectively analyzed the clinical data of 17 patients with psoriasis-associated APL diagnosed and treated in our hospital in the past decade.The 17 patients accounted for 8.3% of the total patients diagnosed with de novo APL during the same period in our hospital. Their clinical characteristics of APL were similar to those of general APL. Four patients had a definite history of taking bimolane. All patients received arsenic trioxide (ATO)-based remission induction and postremission treatment. After induction, 15 patients (88%) achieved hematologic complete remission. With a median follow-up of 27 months, the 3-year estimates of overall survival were 77.2% ± 12.4% and the 3-year estimates of event-free survival were 70.6% ± 13.5%. In addition, the ATO-based remission induction and postremission treatment significantly improved psoriasis symptoms in 83 and 85.7% of patients, respectively. Through the final follow-up, no chronic arsenicosis or secondary malignancy was observed.Psoriasis patients are at high risk for APL. The increased risk is most likely associated with the genetic background and bimolane treatment. The ATO-based therapy is especially suitable for patients with psoriasis-associated APL. Our study also brings a new treatment option for psoriasis.

Objective Hippocampal sclerosis (HS) is the most prevalent pathology in temporal lobe epilepsy (TLE) characterized by segmental neuronal cell loss in the cornu ammonis (CA) 1–4. In addition, migration of granule cells and reorganization of their axons is observed, known as granule cell dispersion (GCD) and mossy fiber sprouting (MFS). The loss of mossy fibers` (MF) target cells in CA4 and CA3 was considered to be causative for MFS. The ILAE HS (International League Against Epilepsy) classification identifies three subtypes with different cell loss patterns in CA1-4. We studied the relation of ILAE HS subtypes to GCD and MFS to corroborate clinico-pathological subgroups in a large retrospective single-center series. Material and methods Hippocampal specimen of 319 patients were screened, 214 could be used for analysis. Immunohistochemical stainings for semi-quantitative analysis of neuronal cell loss (NeuN) and MFS (synaptoporin) were performed. Presurgical data were available from patient files and seizure outcome was classified according to Engel score after surgery. Results In 39 patients (18%) no neuronal cell loss (ILAE no-HS), no GCD and no MFS was observed. In 154 patients (72%) severe neuronal cell loss was seen in CA1, CA4 and CA3 (ILAE HS 1, typical HS); in addition extensive GCD and MFS was observed. In 17 patients (8%) cell loss was seen predominantly in CA1 (ILAE HS 2); despite different cell loss pattern these hippocampi also showed GCD and MFS. In 4 patients (2%) cell loss was predominately detected in CA3 and CA4 (ILAE HS type 3), consecutively GCD and MFS were observed. Longer epilepsy duration and younger age at surgery was more often associated with ILAE HS 2 and febrile convulsions were completely absent in ILAE no-HS. Yet, seizure onset, age at initial precipitating injury and postsurgical seizure outcome did not show any significant association with ILAE HS subtypes. Conclusion GCD and MFS might develop independently from the neuronal cell loss of MF target cells.

Cholestasis is defined as a functional impairment of bile secretion which results in the accumulation of bile acids (BAs) and other toxic molecules in the blood and liver, however, there are very few effective therapies for cholestasis. The farnesoid X receptor (FXR), as a nuclear receptor for BAs, is important in the regulation of BA levels in enterohepatic circulation. It has previously been demonstrated that activation of the FXR pathway may be a useful strategy with which to treat cholestasis. Resveratrol, one of the important ingredients from grape skins and Chinese medicine Polygonum cuspidatum, resulted in FXR‑activated effects in vitro and exhibited a protective effect against α‑naphthylisothiocyanate (ANIT)‑induced cholestasis through FXR regulation in vivo. The underlying mechanisms of resveratrol against ANIT‑induced cholestasis may be due to the regulation of BA homeostasis, improvement of liver injury and attenuation of the inflammatory response, which were regulated in a FXR‑dependent manner and in turn contributed to overall cholestasis alleviation. Overall, resveratrol as a FXR agonist may act as a potential compound for the treatment of drug‑induced cholestasis.

Matrix metalloproteinase-14 (MMP-14) has been demonstrated to play an important role in tumor progression. The aim of this study was to analyze the correlation between MMP-14 expression and clinicopathologic features and its prognostic significance in non-small cell lung cancer (NSCLC). Immunohistochemical staining for MMP-14 protein was performed in 104 patients with NSCLC. High levels of MMP-14 protein were positively correlated with the status of clinical stage (I-II vs. III-IV; P

Cytokine-induced killer (CIK) cells have been shown to be an effective immunotherapy for malignancies. However, their clinical application has been limited due to lack of knowledge on their in vivo kinesis. In this study, we explored their biodistribution by labeling CIK cells with (18)F-FDG and tracking their in vivo migration by PET/CT imaging. In the nine refractory APL patients enrolled in this study, pre-treatment PET/CT scans revealed leukemia burdens in vertebrae, and the bones of the pelvis and limbs. Post-treatment serial PET/CT tracked the localization of CIK cells over time: at 1 h, the majority of these cells accumulated diffusely in the lungs, while the first minor cell activities were observed in brain, liver and spleen; at 4 and 8 h, they not only migrated to the heart, spleen, and liver, but also showed tendencies to accumulate in bone marrow and brain. This specific cell migration route suggested that CIK cells show in vivo functional kinesis and potency as a targeted immunotherapy. The clinical outcome of this small cohort of nine patients supported the efficacy of this regimen: two patients achieved rapid complete remission after three-cycle treatment, and six patients remained stable, subsequently became sensitive to conventional therapy, and also achieved complete remission.

β-amyloid precursor protein (β-APP), also known as Aβ peptide, has a key role in the pathogenesis of Alzheimer's disease, and is also likely to be involved in the development of refractory epilepsy. The mechanism behind the association between β-APP and refractory epilepsy remains to be elucidated. The aim of the present study was to examine the levels of APP mRNA and β-APP protein in patients with refractory epilepsy. Tissue samples were obtained from patients with chronic pharmacoresistant epilepsy who underwent surgery. Levels of APP mRNA and β-APP protein in epileptic temporal lobe and hippocampal tissue were assessed using quantitative polymerase chain reaction, immunohistochemistry and immunofluorescence. The expression levels of protein significantly increased in the temporal cortex and the hippocampus of the patients with epilepsy. β-APP may thus contribute to the pathogenesis of refractory epilepsy.

'Targeting peptides' have demonstrated their value in diagnostic imaging and therapy and novel peptide probes specific to cervical cancer were developed. In the M13KE phage dodecapeptide (12-mer) peptide library, the phage clone S7 showed the best binding to the cancer cells as confirmed by immunofluorescence and flow cytometry assays, and was selected for continued studies. Its binding peptide, CSP3, was synthesized from the sequence of S7's 12-mer at the N-terminus of the minor coat protein pIII of this M13KE phage vector. The peptide's binding was analyzed by the same assays used for S7. It was also assessed using competitive inhibition and binding to a tissue chip. The results demonstrated that the CSP3 peptide bound to cervical carcinoma cells with high sensitivity and specificity. The positive results indicated that the peptide CSP3, conjugated with nanomaterials and chemotherapeutics, may be developed as a targeting vehicle for therapeutic drug delivery against cervical cancer, especially cervical cancer with multiple drug resistance. For this aim, we prepared a CSP3 conjugated liposome drug delivery system containing doxorubicin (DOX) and microRNA101 (miR101) expression plasmids (CSP3-Lipo-DOX-miR101), and the primary result showed that the system demonstrated significantly enhanced cytotoxicity to SiHa cells and DOX resistant SiHa cells, SiHa/ADR. Our results showed that CSP3 is a cervical cancer targeting 12aa peptide with high specificity and sensitivity, and the CSP3 conjugated drug delivery system, CSP3-Lipo-DOX-miR101 has promising potential for development as an efficient drug system for the therapy of cervical cancer.

Obesity and its major co-morbidity, type 2 diabetes, have reached an alarming epidemic prevalence without an effective treatment available. It has been demonstrated that inhibition of SREBP pathway may be a useful strategy to treat obesity with type 2 diabetes. Sterol regulatory element-binding proteins (SREBPs) are major transcription factors regulating the expression of genes involved in biosynthesis of cholesterol, fatty acid and triglyceride. In current study, we identified a small molecule, curcumin, inhibited the SREBP expression in vitro. The inhibition of SREBP by curcumin decreased the biosynthesis of cholesterol and fatty acid. In vivo, curcumin ameliorated HFD-induced body weight gain and fat accumulation in liver or adipose tissues, and improved serum lipid levels and insulin sensitivity in HFD-induced obese mice. Consistently, curcumin regulates SREBPs target genes and metabolism associated genes in liver or adipose tissues, which may directly contribute to the lower lipid level and improvement of insulin resistance. Take together, curcumin, a major active component of Curcuma longa could be a potential leading compound for development of drugs for the prevention of obesity and insulin resistance.

BACKGROUND: The prognosis of acute promyelocytic leukemia (APL) in the elderly is poorer than that of younger patients after treatment with all-trans retinoic acid plus chemotherapy, which is the current standard therapy for APL. A significantly higher mortality during consolidation therapy was found, which is mainly due to deaths from sepsis following chemotherapy-induced myelosuppression. METHODS: A total of 33 patients aged 60 years or older with de novo APL were treated with single-agent arsenic trioxide (ATO) for remission induction and postremission therapy. The postremission therapy continued for up to 4 years. RESULTS: Twenty-nine patients (87.9%) achieved a hematologic complete remission, and the most common adverse event during remission induction was leukocytosis (63.6%). Definite differentiation syndrome was observed in 5 patients. Nonhematologic adverse events were all manageable and reversible. Twenty-eight patients proceeded to postremission therapy. Adverse effects during postremission therapy were mild, transient, and no treatment was required. No patients died from ATO-related toxicities. With a median follow-up of 99 months, the 10-year cumulative incidence of relapse, overall survival, disease-free survival, and cause-specific survival were 10.3%, 69.3%, 64.8%, and 84.8%, respectively, which are comparable with those in the younger APL partners. No significant risks for development of chronic arsenicosis or second malignancy were observed during the follow-up period. CONCLUSIONS: The results indicate that the single-agent ATO regimen is safe and effective with long-term durable remission, and could be used as first-line treatment for elderly patients with de novo APL. Cancer 2013. © 2012 American Cancer Society.

Clear cell renal cell carcinoma (ccRCC) is the most common type of cancer of the renal parenchyma. MicroRNAs (miRNAs) are non-coding RNAs of ~22 nucleotides in length, which function as post‑transcriptional regulators. Recently, the downregulation of miRNA (miR)-492 was observed to be associated with ccRCC; however, the molecular mechanism by which miR492 inhibited ccRCC remained to be elucidated. In the present study, it was demonstrated that miR-492 was markedly downregulated in ccRCC tissues when compared with adjacent normal tissues, as determined by reverse transcription-quantitative poymerase chain reaction (PCR). This downregulation was predominantly due to the hypermethylation of the CpG island of the miR-492 promoter, which was detected by methylation specific PCR and bisulfite genomic sequencing PCR, and was shown to inhibit miR-492 transcription. Through the use of a DNA demethylation agent, 5-aza-2'-deoxycytidine or the histone deacetylase inhibitor 4-phenylbutyric acid, the expression level of miR-492 was significantly upregulated in ccRCC cells, which further inhibited cell proliferation and invasion, while promoting cell apoptosis and adhesion. In conclusion, the present study provided novel insights into the potential mechanisms involved in ccRCC and it is hypothesized that miR-492 may become a promising therapeutic agent in the treatment of ccRCC.

Large tumor suppressor 2 (LATS2) plays significant roles in tumorigenesis and cancer progression. This study was aimed to analyze the correlation between LATS2 expression and clinicopathologic features and its prognostic significance in non-small cell lung cancer (NSCLC). LATS2 expression was examined in 73 NSCLC clinical specimens and 22 normal lung tissues using immunohistochemistry. Low levels of LATS2 protein were inversely associated with the T classification (P=0.001), N classification (P=0.005) and clinical stage (P=0.001) in NSCLC patients. Patients with lower LATS2 expression had a significantly shorter overall survival than patients with high LATS2 expression. Multivariate analysis suggested that low expression of LATS2 was an independent prognostic indicator (P=0.002) for the survival of patients with NSCLC. Furthermore, overexpression of LATS2 resulted in mobility inhibition in NSCLC cell lines A549 and H1299, and reduced protein level of matrix metalloproteinase-2 (MMP-2) and MMP-9. On the contrary, LATS2 siRNA treatment enhanced cell mobility and increased MMP-2 and MMP-9 protein expression level. In conclusion, low expression of LATS2 is a potential unfavorable prognostic factor and promoted cell invasion and migration in NSCLC.

The aim of this study was to determine the efficacy and safety of treatment of pediatric acute promyelocytic leukemia (APL) with single-agent arsenic trioxide (ATO). A total of 19 children (≤ 15 years of age) with newly diagnosed APL were treated with single-agent ATO for remission induction and postremission therapy. Seventeen of the children (89.5%) achieved complete hematologic remission, and 2 early deaths occurred from intracranial hemorrhage. ATO-induced leukocytosis was observed in 13 (68.4%) patients. Other ATO-related toxicities were minimal and transient. Postremission ATO therapy continued for 3 years; the most common side effect was ATO-induced neutropenia. With a median follow-up of 53 months (range, 23-76 months), the calculated 5-year overall survival and event-free survival were 83.9% and 72.7%, respectively, which are comparable with results achieved by the use of ATRA plus chemotherapy, which is the standard therapy for APL. No chronic arsenic toxicity or second malignancies were found during the follow-up period, and arsenic retention was not significant in patients off treatment more than 24 months. ATO resistance was observed in only 1 patient with a complex karyotype. The results indicate the high efficacy and safety of single-agent ATO regimens in the treatment of children with de novo APL.

Background Let-7a has been shown to play important roles in nasopharyngeal carcinoma (NPC) cell proliferation and apoptosis, but little is known about the function and mechanism of let-7a in nasopharyngeal carcinoma metastasis. We aimed to investigate the function and mechanism of let-7a in nasopharyngeal carcinoma metastasis and clarified the regulation of high mobility group A2 (HMGA2) by let-7a. Methods The expression levels of let-7a and HMGA2 were examined in NPC clinical specimens using quantitative reverse transcription-PCR (RT-qPCR). HMGA2 was confirmed as a target of let-7a through luciferase reporter assays, RT-qPCR, and Western blotting. Furthermore, the roles of let-7a and HMGA2 in regulating NPC cells biological properties including proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) process were analyzed with let-7a mimics and si-HMGA2 transfected cells. Results Our study demonstrated that let-7a was downregulated and inversely associated with the clinical stage, T classification and N classification, and HMGA2 was upregulated and directly associated with the clinical stage and N classification in patients with NPC. Moreover, there was an inverse correlation between let-7a expression and HMGA2 expression in NPC patient. In addition, HMGA2 was negatively regulated at the posttranscriptional level by let-7a via a binding site of HMGA2-3′UTR. In addition, synthetic let-7a mimics suppressed NPC cells migration, invasion and EMT process and knockdown of HMGA2 was consistent with the effects of let-7a in NPC cells. Conclusion Let-7a directly downregulates HMGA2 protein expression, which suppress NPC cell migration, invasion and EMT process. Let-7a could serve as a potential diagnostic marker and therapeutic target for NPC. Electronic supplementary material The online version of this article (doi:10.1186/s12967-015-0462-8) contains supplementary material, which is available to authorized users.

To the editor: Approximately 3% to 5% of patients with acute promyelocytic leukemia (APL) will have an extramedullary relapse in their lifetimes, most commonly in the central nervous system (CNS).1 CNS relapse can be isolated or associated with a bone marrow (BM) relapse. The most appropriate clinical management remains controversial.2 The major obstacle to all treatments is the need for the therapeutic drugs to penetrate the blood-brain barrier (BBB). Although arsenic trioxide (ATO) and all-trans retinoic acid (ATRA) are among the frontline medications for APL intramarrow treatment, these water-soluble medicines have limited ability to cross the BBB and cannot reach therapeutically effective levels in the cerbrospinal fluid (CSF). With regular oral doses, the ATO level in CSF has been reported to reach only 17.7% of the corresponding levels in plasma.3 Previously, we investigated the potential efficacy of combination therapy with mannitol and ATO using a rabbit model, and we showed that the approach caused a transient increase in the BBB permeability for ATO, thereby increasing the ATO concentration in CSF.4 Our experiments using human cortical neurons revealed a differential tolerance of APL blasts to different concentrations of ATO.5,6 We also identified a safe range of ATO concentrations in the human CNS.7,8 The results from these collective studies allowed an ATO concentration in CSF that was both safe and therapeutic to be achieved.9 Here, we describe our efforts to extend the application of our method to additional patients. This study was registered at www.isrctn.org (#ISRCTN94954912). The study protocol was reviewed and approved by the Harbin Medical University Medical Ethics Committee, and signed informed consent was obtained from all patients or their legal guardians. Seventeen CNS APL patients diagnosed between 2000 and 2010 were included in this study (10 males and 7 females, between 6 and 50 years old). Expression of the PML/RARα gene [or t(15;17)(q22;q21) transcripts] was detected in all patient CSF. For all patients, the induction treatment was started immediately on diagnosis. The daily protocol was as follows: a bolus infusion of 125 mL of 20% mannitol mixed in 100 mL normal saline (NS) (for children ≤15 years old: 50 mL mannitol and 50 mL NS) was administered intravenously at a flow rate of 12 ∼20 mL/min (∼8-11 minutes total), followed by a slow intravenous infusion of 125 mL of 20% mannitol plus 7.0 mg/m2/day ATO (for children ≤15 years: 50 mL mannitol and 0.16 mg/kg/day ATO) in 500 mL NS at a flow rate of 1.0 mL/min (∼9-10.5 hours total). Patients were instructed to rest in bed during the entire procedure. Urine flow was measured to ensure maintenance of a rate of at least 30 to 50 mL/h. Daily infusions were continued until the patient’s CSF was found to be free of APL blasts/promyelocytes. The level of elemental arsenic metabolites was measured in each patient’s CSF and blood samples, which had been taken immediately after every other day’s infusion. Consolidation therapy started 2 weeks after the induction therapy and was repeated at 2-week intervals. A total of 3 cycles of consolidation therapy was given to all the patients who achieved normal CSF morphology with induction treatment. For each cycle of consolidation, daily ATO and mannitol infusion (administered as described for the induction treatment) continued for 14 consecutive days. Absence of PML/RARα transcripts in CSF and BM (defined as complete molecular remission [CMR]) was achieved prior to commencement of lifetime maintenance therapy. The first year after CMR, a 14-day cycle administration of ATO and mannitol was repeated at 1-month intervals. In the second year, a protocol identical to that of year 1 was instituted, but using a 3-month interval regimen. From the third year on, the protocol was administered at 6-month intervals and maintained for life. PML/RARα levels were monitored at 6-month intervals. If a relapse was identified, another cycle of remission induction was started. Clinical monitoring and supportive treatment followed our departmental guidelines.10 In 16 of the 17 patients examined, abnormal blasts/promyelocytes from CSF were eliminated in 18 to 32 days (median, 24 days) after the start of induction treatment. All the patients tolerated the induction well. There were no complaints of side effects associated with the use of mannitol. There were no documented cases of acute arsenic poisoning. No patients discontinued use of ATO during the induction. There were some cases of mild liver enzyme changes and mild nonhematologic toxicity associated with ATO, but in all instances, these abnormalities resolved with the use of hydroxyurea and dexamethasone for 3 to 5 days. There were no significant differences in the level of arsenic metabolites between children and adults. The correlation between parallel blood and CSF arsenic levels was significant (r = 0.998). Over the course of the entire induction treatment process, the concentrations of arsenic in the blood and CSF were fairly stable in each patient (Figure 1). For each individual, the arsenic level in CSF was ∼99.7% of those in the paired blood samples. However, arsenic levels in different individuals were highly variable in blood and CSF. Of the 16 patients who responded to induction therapy, all achieved CMR after the first consolidation cycle. Nine of these patients retained their CMR status during the follow-up period (median, 92 months; range, 70-125 months). Among the other 7 patients, there were 1 to 3 relapses in BM and/or CNS after their first CMR. All 7 of these patients responded well to ATO and mannitol during reinduction: 5 of them achieved CMR again and stayed free of relapse; 2 died after withdrawing treatment 3 to 5 months after their second or third remission. One 6-year-old female patient was nonresponsive to induction treatment after 49 days and later received ATRA and ATO alternative therapy. She died 15 months after the start of therapy. Eventually, the mean 5-year overall survival rate for all 17 patients was 82.4%. The mean 5-year event-free survival rate for all patients was 52.9%. At the time of the last follow-up appointment, all patients remained in generally good health without signs of chronic arsenic intoxication or second malignancy. Figure 1 Elemental arsenic levels in patient-paired blood and CSF samples measured during induction treatment.

Sterol regulatory element-binding proteins (SREBPs) are major transcription factors regulating the expression of genes involved in biosynthesis of cholesterol, fatty acids, and triglycerides. We investigated the effect of the specific SREBP suppressor andrographolide, a natural compound isolated from Andrographis paniculata, on the regulation of SREBP signaling by use of Western blot, reporter gene assay, and quantitative real-time polymerase chain reaction analysis. In addition, the antiobesity effects of andrographolide were evaluated in C57BL/6 mice with high-fat diet (HFD)-induced obesity. Our results showed that andrographolide downregulated the expressions of SREBPs target genes and decreased cellular lipid accumulation in vitro. Further, andrographolide (100 mg/kg per day) attenuated HFD-induced body weight gain and fat accumulation in liver or adipose tissues, and improved serum lipid levels and insulin or glucose sensitivity in HFD-induced obese mice. Andrographolide effectively suppressed the respiratory quotient, energy expenditure, and oxygen consumption, which may have contributed to the decreased body-weight gain of the obese mice fed with a HFD. Consistently, andrographolide regulated SREBP target genes and metabolism-associated genes in liver or brown adipose tissue, which may have directly contributed to the lower lipid levels and enhanced insulin sensitivity. Taken together, our results indicated that andrographolide ameliorated lipid metabolism and improved glucose use in mice with HFD-induced obesity. Andrographolide has potential as a leading compound in the prevention or treatment of obesity and insulin resistance.

To clarify the anatomical organization of human memory remains a major challenge in clinical neuroscience. Experimental data suggest dentate gyrus granule cells play a major role in memory acquisition, i.e. pattern separation and rapid pattern completion, whereas hippocampal CA1 neurons are implicated in place memory and autobiographical memory retrieval. Patients with temporal lobe epilepsy present with a broad spectrum of memory impairment, which can be assessed during clinical examination. Although long seizure histories may contribute to a pathophysiological reorganization of functional connectivity, surgical resection of the epileptic hippocampus offers a unique possibility to anatomically study the differential contribution of hippocampal subfields to compromised learning and memory in humans. Herein, we tested the hypothesis of hippocampal subfield specialization in a series of 100 consecutive patients with temporal lobe epilepsy submitted to epilepsy surgery. Memory profiles were obtained from intracarotid amobarbital testing and non-invasive verbal memory assessment before surgery, and correlated with histopathologically quantified cell loss pattern in hippocampal subfields obtained from the same patients using the new international consensus classification for hippocampal sclerosis proposed by the International League against Epilepsy (HS ILAE). Interestingly, patients with CA1 predominant cell loss (HS ILAE Type 2; n = 13) did not show declarative memory impairment and were indistinguishable from patients without any hippocampal cell loss (n = 19). In contrast, 63 patients with neuronal loss affecting all hippocampal subfields including CA1, CA4 and dentate gyrus (HS ILAE Type 1), or predominant cell loss in CA4 and partially affecting also CA3 and dentate gyrus (HS ILAE Type 3, n = 5) showed significantly reduced declarative memory capacities (intracarotid amobarbital testing: P < 0.001; verbal memory: P < 0.05). Our results suggested an alternative model of how memory processing can be organized amongst hippocampal subfields, and that CA1 pyramidal cells are less critically involved in declarative human memory acquisition compared to dentate gyrus granule cells or CA4/CA3 pyramidal cells.

Uveal melanoma arises in the eye, and it spreads to distant organs in almost half of patients, leading to a fatal outcome. To metastasize, uveal melanoma cells must transmigrate into and out of the microvasculature, crossing the monolayer of endothelial cells that separates the vessel lumen from surrounding tissues. We investigated how human uveal melanoma cells cross the endothelial cell monolayer, using a cultured cell system with primary human endothelial cell monolayers on hydrogel substrates. We found that uveal melanoma cells transmigrate by a novel and unexpected mechanism. Uveal melanoma cells intercalate into the endothelial cell monolayer and flatten out, assuming a shape and geometry similar to those of endothelial cells in the monolayer. After an extended period of time in the intercalated state, the uveal melanoma cells round up and migrate underneath the monolayer. VCAM is present on endothelial cells, and anti-VCAM antibodies slowed the process of intercalation. Depletion of BAP1, a known suppressor of metastasis in patients, increased the amount of transmigration of uveal melanoma cells in transwell assays; but BAP1 depletion did not affect the rate of intercalation, based on movies of living cells. Our results reveal a novel route of transendothelial migration for uveal melanoma cells, and they provide insight into the mechanism by which loss of BAP1 promotes metastasis.

Psychogenic non-epileptic seizures (PNES) are paroxysmal behaviors that resemble epileptic seizures but lack abnormal electrical activity. Recent studies suggest aberrant functional connectivity involving specific brain regions in PNES. Little is known, however, about alterations of topological organization of whole-brain functional and structural connectivity networks in PNES. We constructed functional connectivity networks from resting-state functional MRI signal correlations and structural connectivity networks from diffusion tensor imaging tractography in 17 PNES patients and 20 healthy controls. Graph theoretical analysis was employed to compute network properties. Moreover, we investigated the relationship between functional and structural connectivity networks. We found that PNES patients exhibited altered small-worldness in both functional and structural networks and shifted towards a more regular (lattice-like) organization, which could serve as a potential imaging biomarker for PNES. In addition, many regional characteristics were altered in structural connectivity network, involving attention, sensorimotor, subcortical and default-mode networks. These regions with altered nodal characteristics likely reflect disease-specific pathophysiology in PNES. Importantly, the coupling strength of functional-structural connectivity was decreased and exhibited high sensitivity and specificity to differentiate PNES patients from healthy controls, suggesting that the decoupling strength of functional-structural connectivity might be an important characteristic reflecting the mechanisms of PNES. This is the first study to explore the altered topological organization in PNES combining functional and structural connectivity networks, providing a new way to understand the pathophysiological mechanisms of PNES.

Curcumin, a principle bioactive component of Curcuma longa L, is well known for its anti-hyperlipidemia effect. However, no holistic metabolic information of curcumin on hyperlipidemia models has been revealed, which may provide us an insight into the underlying mechanism. In the present work, NMR and MS based metabolomics was conducted to investigate the intervention effect of curcumin on hyperlipidemia mice induced by high-fat diet (HFD) feeding for 12 weeks. The HFD induced animals were orally administered with curcumin (40, 80 mg/kg) or lovastatin (30 mg/kg, positive control) once a day during the inducing period. Serum biochemistry assay of TC, TG, LDL-c, and HDL-c was conducted and proved that treatment of curcumin or lovastatin can significantly improve the lipid profiles. Subsequently, metabolomics analysis was carried out for urine samples. Orthogonal Partial Least Squares-Discriminant analysis (OPLS-DA) was employed to investigate the anti-hyperlipidemia effect of curcumin and to detect related potential biomarkers. Totally, 35 biomarkers were identified, including 31 by NMR and nine by MS (five by both). It turned out that curcumin treatment can partially recover the metabolism disorders induced by HFD, with the following metabolic pathways involved: TCA cycle, glycolysis and gluconeogenesis, synthesis of ketone bodies and cholesterol, ketogenesis of branched chain amino acid, choline metabolism, and fatty acid metabolism. Besides, NMR and MS based metabolomics proved to be powerful tools in investigating pharmacodynamics effect of natural products and underlying mechanisms.

Background: Bipolar disorder affects approximately 1% of the population at an annual cost of $45 billion in the US. Estimates of non-compliance with mood stabilizer therapy range as high as 64%. The objective of this study was to document the use patterns with mood stabilizers achieved by patients with bipolar disorder and to estimate the direct health care costs associated with sub-optimal drug therapy. Methods: Paid claims for 3349 California Medicaid patients with bipolar disorder were used to document the use patterns for mood stabilizers achieved by patients with bipolar disorder. The impact of the patient’s drug use patterns on likelihood of antipsychotic or antidepressant use within 1 year and health care costs incurred during the first posttreatment year were also estimated. Results: Only 42.4% of patients used a mood stabilizer during the first posttreatment year; over 60% of treated patients switch or augment their initial therapy within 1 year, and only 5.5% of patients used a mood stabilizer consistently for 1 year. Direct health care costs were significantly higher among those patients who delayed or did not use mood-stabilizing agents during the first year. Limitations: Medi-Cal covers poor and disabled patients and is not representative of the general population. Paid claims data do not include clinical markers for severity of illness or treatment response. Conclusions: Suboptimal use patterns for mood stabilizing medications were frequent and costly. Strategies to improve compliance with mood stabilizer regimens, along with new treatment options, are needed to improve treatment outcomes.

Diabetic cardiomyopathy is a common cardiac condition in patients with diabetes mellitus, which can result in cardiac hypertrophy and subsequent heart failure, associated with pyroptosis, the pro-inflammatory programmed cell death. MicroRNAs (miRNAs), small endogenous non-coding RNAs, have been shown to be involved in diabetic cardiomyopathy. However, whether miRNAs regulate pyroptosis in diabetic cardiomyopathy remains unknown. Our study revealed that mir-30d expression was substantially increased in streptozotocin (STZ)-induced diabetic rats and in high-glucose-treated cardiomyocytes as well. Upregulation of mir-30d promoted cardiomyocyte pyroptosis in diabetic cardiomyopathy; conversely, knockdown of mir-30d attenuated it. In an effort to understand the signaling mechanisms underlying the pro-pyroptotic property of mir-30d, we found that forced expression of mir-30d upregulated caspase-1 and pro-inflammatory cytokines IL-1β and IL-18. Moreover, mir-30d directly repressed foxo3a expression and its downstream protein, apoptosis repressor with caspase recruitment domain (ARC). Furthermore, silencing ARC by siRNA mimicked the action of mir-30d: upregulating caspase-1 and inducing pyroptosis. These findings promoted us to propose a new signaling pathway leading to cardiomyocyte pyroptosis under hyperglycemic conditions: mir-30d↑→foxo3a↓→ ARC↓→caspase-1↑→IL-1β, IL-18↑→pyroptosis↑. Therefore, mir-30d may be a promising therapeutic target for the management of diabetic cardiomyopathy.

Uveal melanoma, cancer arising from the pigmented uveal layer of the eye, is highly metastatic despite being limited to spreading only through the bloodstream. Almost half of patients develop distant metastatic disease, most often in the liver, even after the tumor-bearing eye is completely removed. The key to understanding and targeting uveal melanoma metastasis is in understanding how circulating tumors cells exit the bloodstream at distant sites of metastasis, and then invade and colonize distant organs. To address the first process, transendothelial migration, we modeled the blood vessel wall using primary human microvascular endothelial monolayers grown on polyacrylamide substrates that mimic the physiological stiffness of normal tissue. To these monolayers, we added 92.1 and OCM-1A uveal melanoma cells, and then collected time-lapse movies using spinning-disc confocal and differential interference contrast microscopy to follow the transmigration of the uveal melanoma cells. We found that both cell lines transmigrate through endothelial monolayers by a route distinctly different from the paracellular migration we see for trafficking immune cells. During this multistep process, uveal melanoma cells first identify paracellular junctions and then intercalate between endothelial cells, taking on a flattened morphology and maintaining contacts with their adjacent endothelial cells; reminiscent of vasculogenic mimicry. Tumor cells then extend invasive projections beneath adjacent endothelial cells, before eventually migrate under the endothelium. Immunofluorescence revealed cortical actin networks with strong cortactin staining within these invasive projections. To visualize these structures better, we transfected uveal melanoma cells with F-tractin, a fluorescent fusion protein that specifically binds filamentous actin without disrupting actin function, that allows imaging of actin dynamics in live cells. Time-lapse confocal images revealed complex actin-rich projections invading and sampling the interface between the endothelial monolayer and the substrate. Dynamic actin cytoskeletal reorganization was also apparent as cells migrated under the monolayer. Previous studies have linked inactivating mutations in BAP1 to metastatic spread of uveal melanoma in patients. To test whether BAP1 loss plays a role in transendothelial migration, we knocked down BAP1 expression in our uveal melanoma cell lines. We found that BAP1-depleted cells transmigrated significantly faster through endothelial monolayers in transwell assays than control knockdown cells. Time-lapse movies showed no significant differences in the first steps of initiation or intercalation between BAP1 knockdown and control cells. These studies identify a novel mechanism of transendothelial migration in uveal melanoma cells and suggest that BAP1 depletion specifically affects the ability of cells to invade under and exit the monolayer. Citation Format: Michael D. Onken, Olivia L. Mooren, Jinmei Li, John A. Cooper. Transendothelial migration of uveal melanoma cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3163. doi:10.1158/1538-7445.AM2014-3163

Acute promyelocytic leukemia (APL) causes coagulation, which can worsen following initiation of chemotherapy and is improved or corrected by the differentiating agents, all trans retinoic acid (ATRA) and arsenic trioxide (As2O3). The relationship of procoagulant activity to phosphatidylserine (PS) exposure on leukemic cells has not been clarified, although prior studies indicate that APL cells expose tissue factor (TF). Procoagulant activity of leukemic cells was measured in a modified prothrombin time in which leukemic cells replaced thromboplastin. The presence of phosphatidylserine (PS) in neutrophils and mononuclear cells (MNC) from healthy donors and APL cells from patients was investigated by flow-cytometry and confocal microscopy. The assembly of extrinsic tenase, intrinsic tenase and prothrombinase complexes on these cells was studied using enzymatic assays employing plasma or purified proteins. Lactadherin was utilized as a probe to track PS exposure and as an agent to block exposed PS. Isolated APL cells exhibited patches that stained with lactadherin, but neutrophils and MNC did not, indicating more PS exposure on APL cells than the other two cell types. Exclusion of propidium iodide by the leukemic cells indicated that PS exposure occurs in the absence of frank apoptosis. Coagulant activity increased approx 20-fold after cells were exposed to 0.1 μM daunorubicin for 24 hr with 70% of apoptosis and decreased by 85% after 24 hr treatment with 1 μM ATRA or As2O3 with partial differentiation. Inhibition of procoagulant activity by ATRA and As2O3 corresponded to decreased PS and decrease activity for all three enzyme complexes. Lactadherin, which blocks PS binding sites, inhibited Xase and prothrombin conversion by their respective APL-assembled activating complexes. APL cells support all procoagulant reactions leading to robust thrombin formation. This ability results from concomitant PS and TF exposure at the outer leaflet of cell membrane. In contrast, less PS on neutrophils and MNC do not support the coagulation.