Plasma exosomal microRNAs are non-invasive biomarkers of moyamoya disease: A pilot study

Abstract Background As a progressive cerebrovascular disease, Moyamoya Disease (MMD) is a common cause of stroke in children and adults. However, the early biomarkers and pathogenesis of MMD remain poorly understood. Methods and material This study was conducted using plasma exosome samples from MMD patients. Next-generation high-throughput sequencing, real-time quantitative PCR, gene ontology analysis, and Kyoto Encyclopaedia of Genes and Genomes pathway analysis of ideal exosomal miRNAs that could be used as potential biomarkers of MMD were performed. The area under the Receiver Operating Characteristic (ROC) curve was used to evaluate the sensitivity and specificity of biomarkers for predicting events. Results Exosomes were successfully isolated and miRNA-sequence analysis yielded 1,002 differentially expressed miRNAs. Functional analysis revealed that they were mainly enriched in axon guidance, regulation of the actin cytoskeleton and the MAPK signaling pathway. Furthermore, 10 miRNAs (miR-1306-5p, miR-196b-5p, miR-19a-3p, miR-22-3p, miR-320b, miR-34a-5p, miR-485-3p, miR-489-3p, miR-501-3p, and miR-487-3p) were found to be associated with the most sensitive and specific pathways for MMD prediction. Conclusions Several plasma secretory miRNAs closely related to the development of MMD have been identified, which can be used as biomarkers of MMD and contribute to differentiating MMD from non-MMD patients before digital subtraction angiography.

High expression of ZEB1 is associated with EMAST & metastasis in colorectal cancer patients

Background & objectives: Transforming growth factor-beta (TGF-?) signalling pathway has been reported to be involved in metastasis and at the same time has been considered compellingly an important mediator of epithelial-to-mesenchymal transition (EMT). Besides, EMT process is maintained by zinc-finger E-box-binding homeobox 1 (ZEB1) gene which is induced by TGF-? pathway. TGF-? has been shown to be associated with elevated microsatellite alterations at selected tetranucleotide repeats (EMAST) phenomenon, which is one of the prognostic biomarkers of colorectal cancer (CRC). This study was conducted to determine the link among ZEB1-induced TGF-?, EMAST status and metastasis. Methods: The expression level of ZEB1 was evaluated using quantitative reverse transcription (qRT) real-time PCR in 122 formalin fixed paraffin-embedded tissues of CRC sample with known EMAST status and TGF-?/Smad-dependent pathways. The association among ZEB1 expression, TGF-? signalling pathway, EMAST status and metastatic behaviour was examined. Results: ZEB1 gene expression level was higher in tumour tissues as compared to normal samples (P<0.045). In addition, ZEB1 positive expression level was associated significantly with metastasis (P=0.05), EMAST+ status (P=0.052) and activated TGF-? signalling pathway (P=0.002). Interpretation & conclusions: Our results validated significant association between activated TGF-? signalling pathway and EMAST+ phenotype with higher expression of ZEB1 and higher level of metastasis.

Drug discovery of sclerostin inhibitors

Sclerostin, a protein secreted from osteocytes, negatively regulates the WNT signaling pathway by binding to the LRP5/6 co-receptors and further inhibits bone formation and promotes bone resorption. Sclerostin contributes to musculoskeletal system-related diseases, making it a promising therapeutic target for the treatment of WNT-related bone diseases. Additionally, emerging evidence indicates that sclerostin contributes to the development of cancers, obesity, and diabetes, suggesting that it may be a promising therapeutic target for these diseases. Notably, cardiovascular diseases are related to the protective role of sclerostin. In this review, we summarize three distinct types of inhibitors targeting sclerostin, monoclonal antibodies, aptamers, and small-molecule inhibitors, from which monoclonal antibodies have been developed. As the first-in-class sclerostin inhibitor approved by the U.S. FDA, the monoclonal antibody romosozumab has demonstrated excellent effectiveness in the treatment of postmenopausal osteoporosis; however, it conferred high cardiovascular risk in clinical trials. Furthermore, romosozumab could only be administered by injection, which may cause compliance issues for patients who prefer oral therapy. Considering these above safety and compliance concerns, we therefore present relevant discussion and offer perspectives on the development of next-generation sclerostin inhibitors by following several ways, such as concomitant medication, artificial intelligence-based strategy, druggable modification, and bispecific inhibitors strategy.

Chaetocin:a review of its anticancer potential and mechanisms / 中国药理学与毒理学杂志

Chaetocin is a natural metabolite product with various biological activities and pharmacological functions isolated from Chaetomium species fungi belonging to the thiodiketopyrazines. Numerous studies have demonstrated a wide range of antitumor activities of chaetocin in vitro and in vivo. Several studies have demonstrated that chaetocin sup?presses the growth and proliferation of various tumour cells by regulating multiple signalling pathways related to tumour initiation and progression, inducing cancer cell apoptosis (intrinsic and extrinsic), enhancing autophagy, inducing cell cycle arrest, as well as inhibiting tumour angiogenesis, invasion and migration. The antitumor effects and molecular mechanisms of chaetocin are reviewed and analysed in this paper, and the prospective applications of chaetocin in cancer prevention and therapy are also discussed. Our review provides the theoretical basis for exploiting the clinical applica?tion of chaetocin in cancer treatment.

Hepatic protective effects of Shenling Baizhu powder, a herbal compound, against inflammatory damage via TLR4/NLRP3 signalling pathway in rats with nonalcoholic fatty liver disease / 中西医结合学报

OBJECTIVE@#High-fat diet (HFD) and inflammation are two key contributors to nonalcoholic fatty liver disease (NAFLD). Shenling Baizhu powder (SLBZP), a classical herbal compound, has been successfully used to alleviate NAFLD. However, its specific mechanisms are not fully understood. In this study, we assessed the anti-NAFLD effect of SLBZP in vivo.@*METHODS@#Rats were fed an HFD with or without SLBZP or with probiotics. At the end of week 16, an echo magnetic resonance imaging (EchoMRI) body composition analyser was used to quantitatively analyse body composition; a micro-computed tomography (micro-CT) imaging system was used to evaluate whole body and liver fat; and the Moor full-field laser perfusion imager 2 was used to assess liver microcirculation, after which, all rats were sacrificed. Then, biochemical indicators in the blood and the ultrastructure of rat livers were evaluated. Protein expression related to the liver Toll-like receptor 4 (TLR4)/Nod-like receptor family pyrin domain-containing 3 (NLRP3) signalling pathway was assessed using Western blot analysis. Further, high-throughput screening of 29 related inflammatory factors in liver tissue was performed using a cytokine array.@*RESULTS@#SLBZP supplementation reduced body weight, serum free fatty acid, and insulin resistance index (P < 0.05). It also ameliorated liver microcirculation and ultrastructural abnormalities. EchoMRI and micro-CT quantitative analyses showed that treatment with SLBZP reduced fat mass and visceral fat (P < 0.05 and P < 0.01, respectively). In addition, SLBZP decreased the expression of lipopolysaccharide (LPS)-activated TLR4/NLRP3 signalling pathway-related proteins and altered the expression levels of some inflammatory cytokines in liver tissues.@*CONCLUSION@#SLBZP can inhibit NLRP3 inflammasome activation and interleukin-1β release by suppressing LPS-induced TLR4 expression in rats with HFD-induced NAFLD. Thus, SLBZP may be beneficial for the prevention and treatment of inflammatory damage and associated diseases.

Intracellular calcium ions facilitate dengue virus entry into endothelial cells and compromise endothelial barrier integrity

Objective: To investigate the involvement of Ca2+ in dengue virus (DENV)-infected human umbilical vein endothelial cells (HUVECs) and the disruption of endothelial integrity. Methods: HUVECs were infected with DENV-2 in the presence of intracellular Ca2+ or endoplasmic reticulum Ca2+ chelators. Virus infectivity was measured by focus-forming assay and quantitative RT-PCR. Intracellular Ca2+ was measured using Fluo-4-AM dye. VE-cadherin and focal adhesion kinase (FAK) expressions were investigated by immunofluorescence and immunoblotting assays, respectively. Results: DENV infection increased intracellular cytosolic Ca2+ levels and caused disassembly of the adherens junction protein, VEcadherin as evidenced by decreased VE-cadherin expression at the periphery of DENV-2 infected HUVECs. Depletion of intracellular Ca2+ stores, particularly those of the endoplasmic reticulum Ca2+, significantly decreased DENV yield in HUVECs. Decreased virus yield following the depletion of intracellular Ca2+ was caused by the inhibition of viral entry into HUVECs and not the inhibition of viral binding or attachment. DENV-2 infection also resulted in Ca2+- dependent activation of FAK. Conclusions: Intracellular Ca2+ is required for the early phases of DENV infection in endothelial cells. Increased cytosolic Ca2+ levels in endothelial cells during DENV infection activated FAK, disrupted adherens junctions and compromised barrier integrity. Thus, Ca2+ plays an important role in DENV infection in endothelial cells.

Ginsenoside Rb1 Protects Human Umbilical Vein Endothelial Cells against High Glucose-Induced Mitochondria-Related Apoptosis through Activating SIRT3 Signalling Pathway / 中国结合医学杂志

OBJECTIVE@#To investigate whether ginsenoside Rb1 (Rb1) can protect human umbilical vein endothelial cells (HUVECs) against high glucose-induced apoptosis and examine the underlying mechanism.@*METHODS@#HUVECs were divided into 5 groups: control group (5.5 mmol/L glucose), high glucose (HG, 40 mmol/L) treatment group, Rb1 (50 µ mol/L) treatment group, Rb1 plus HG treatment group, and Rb1 and 3-(@*RESULTS@#Rb1 ameliorated survival in cells in which apoptosis was induced by high glucose (P<0.05 or P<0.01). Upon the addition of Rb1, mitochondrial and intracellular reactive oxygen species generation and malondialdehyde levels were decreased (P<0.01), while the activities of antioxidant enzymes were increased (P<0.05 or P<0.01). Rb1 preserved the mitochondrial membrane potential and reduced the release of Cyt-c from the mitochondria into the cytosol (P<0.01). In addition, Rb1 upregulated mitochondrial biogenesis-associated proteins (P<0.01). Notably, the cytoprotective effects of Rb1 were correlated with SIRT3 signalling pathway activation (P<0.01). The effect of Rb1 against high glucose-induced mitochondria-related apoptosis was restrained by 3-TYP (P<0.05 or P<0.01).@*CONCLUSION@#Rb1 could protect HUVECs from high glucose-induced apoptosis by promoting mitochondrial function and suppressing oxidative stress through the SIRT3 signalling pathway.

Freeing the brake: Proliferation needs primary cilium to disassemble

Primary cilia are non-motile, microtubule-based, antennae-like organelle that protrude out from the cell surfaceand perform sensory function or transduce physiological signals in majority of the vertebrate cells. Cilia areassembled on basal bodies that are transformed centrioles. The assembly-disassembly of primary cilia maypose an additional measure on regulating cell cycle in vertebrate cells. While primary cilia are commonly foundin differentiated or quiescent cells that are not cycling, disassembly of primary cilia may promote re-entry ofthese cells into the mitotic cycle, and support proliferation. Many cancer tissues or cancer-derived cells exhibitloss of primary cilia. However, primary cilia may also promote tumorigenesis in some contexts throughgrowth-promoting signalling. This review will shed light on recent advancements of temporal coordination ofciliary disassembly and cell cycle progression, with a focus on how cilia loss may support tumorigenesis invarious epithelial cancers

Improved drug carriage and protective potential against Cisplatin-induced toxicity using Boldine-loaded PLGA nanoparticles

Background: Cisplatin is a widely-used potent anti-cancer drug having severe side-effects precluding itssustained use.Objectives: Poly (lactide-co-glycolide) (PLGA)-nanoparticles loaded Boldine, an antioxidant ingredient ofethanolic extract of Boldo plant (Peumus boldus) was tested in cancer mice model, Mus musculus toexamine if it could reduce unwanted Cisplatin-induced toxicity in normal tissue.Material and methods: Nano-encapsulation of Boldine was done by following the standardized solventdisplacement method. Physico-chemical characterization of PLGA-encapsulated nano-Boldine (NBol) wasaccomplished through analyses of various spectroscopic techniques. Status of major antioxidant enzymes, functional markers, and lipid peroxidation (LPO) was also determined in certain tissue and serumsamples. Percentage of cells undergoing cytotoxic death, Reactive oxygen species (ROS) accumulationand mitochondrial functioning were analyzed in both normal and cancer mice. Nanoscale changes inchromatin organization were assessed by Transmission electron microscopy (TEM). mRNA and proteinexpressions of Top II, Bax, Bcl-2, Cyt c, caspase 3 were studied by RT-PCR, immunoblot andimmunofluorescence.Results: NBol had faster mobility, site-specific action and ability of sustained particle release. NBol readilyentered cells, prevented Cisplatin to intercalate with dsDNA resulting in reduction of chromatincondensation, with corresponding changes in ROS levels, mitochondrial functioning and antioxidantenzyme activities, leading to reduction in Deoxyribose nucleic acid (DNA) damage and cytotoxic celldeath. Expression pattern of apoptotic genes like Top II, p53, Bax, Bcl-2, cytochrome c and caspase-3suggested greater cytoprotective potentials of NBol in normal tissues.Conclusions: Compared to Boldine (Bol), NBol had better ability of drug carriage and protective potentials(29.00% approximately) against Cisplatin-induced toxicity. Combinational therapeutic use of PLGA-NBolcan reduce unwanted Cisplatin-induced cellular toxicity facilitating use of Cisplatin.© 2018 Transdisciplinary University, Bangalore and World Ayurveda Foundation. Publishing Services byElsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Neuroprotective natural products: promising candidates against neurodegenerative diseases / Productos naturales con propiedades neuroprotectivas: posibles candidatos para el tratamiento de enfermedades neurodegenerativas

Neurodegeneration is a progressive loss of neurons both structurally and functionally causing neuronal cell death ultimately leading to development of various neurodegenerative diseases. Due to poor pharmacokinetic profile of neurotrophins, there still remains a challenge in their neurotrophic therapy where plants, bacteria and fungi, as natural products, could act as promising candidates against various neurological disorders by modulating the neurotrophic activity. Therefore, these natural products that mimic neurotrophins, could develop novel therapeutic approaches to herbal drug that can ameliorate neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease and other associated neurological disorders. Taking into account the failure of strategies involving single neurotrophins for the treatment of neurodegenerative diseases, we propose a combination of small molecules of natural products that may work synergistically to restore neuronal functions, minimize side effects and target multiple pathways for a more effective treatment.

La neurodegeneración es una pérdida progresiva de neuronas, tanto estructural como funcional, que causa la muerte neuronal, lo que conduce al desarrollo de diversas enfermedades neurodegenerativas. Debido al pobre perfil farmacocinético de las neurotrofinas, existe un desafío en su terapia neurotrófica donde plantas, bacterias y hongos, como productos naturales, podrían actuar como candidatos contra diversos trastornos neurológicos al modular la actividad neurotrófica. Estos productos naturales que asemejan a las neurotrofinas podrían desarrollar enfoques terapéuticos novedosos como medicamentos a base de hierbas que pueden mejorar enfermedades neurodegenerativas como: Parkinson, Alzheimer y otros trastornos neurológicos asociados. Teniendo en cuenta el fracaso de las estrategias terapéuticas de neurotrofinas para las enfermedades neurodegenerativas, proponemos una combinación de pequeñas moléculas de productos naturales que pueden funcionar sinérgicamente para restaurar las funciones neuronales, minimizar los efectos secundarios y apuntar a múltiples vías para un tratamiento más efectivo.

Role of tissue engineering in oral & maxillofacial surgery – a review

Tissue engineering can be defined as the “reconstitution of tissue and organs, in vitro for use as model systems in basic and applied research, or for use as grafts to replace damaged or diseased body parts or body functions”. Biomaterials have been used as replacement tissues and grafts have been used to reconstruct defects in craniofacial region till Uristmade the first attempt of producing exogenous bone with the help of bone morphogenetic proteins. The success of tissue engineering over the field of all transplantation is that conceptually a three-dimensional functional tissue is designed. This field has become a boon to the Cranio Maxillofacial surgeons and has provided them with a supplement to existing treatment for reconstruction of Oral & Craniofacial region. The purpose of this article is to present an overview of the various uses of tissue engineering in the field of Oral and Maxillofacial Surgery.

Notch Signaling Promotes Proliferation and Migration of SW982 Synovial Sarcoma Cells / 中国医科大学学报

Objective To investigate the effect of the Notch signaling pathway on the proliferation and invasion of human SW982 synovial sarcoma cells. Methods SW982 cells and normal human synovial cells were routinely cultured, and the expression of proteins related to the Notch pathway was compared. The Notch signaling pathway was manipulated by NICD1 overexpression, CFB1 shRNA lentivirus, and the γ-secretase inhibitor, DAPT. CCK-8 and wound healing assays were carried out to investigate the role of the Notch signaling pathway in SW982 cells. Results The Notch signaling pathway clearly showed higher activity in human SW982 synovial sarcoma cells than in normal human synovial cells (P ＜ 0.05). The proliferation and invasion of SW982 cells were significantly upregulated by overexpressing NICD1; however, were suppressed by downregulating the Notch signaling pathway using CFB1 shRNA or DAPT (P ＜ 0.05). Conclusion Our findings demonstrate that the proliferation and invasion of human SW982 synovial sarcoma cells are dependent on Notch signaling pathway activity.

Neurological Disorder Brain Model: A Lesson from Marine Worms (Annelida: Polychaeta)

@#The incidence of neurodegenerative diseases is directly proportional to age. The prevalence of non-communicable diseases, for example, Alzheimer’s and Parkinson’s, is expected to rise in the coming years. Understanding the etiopathology of these diseases is a crucial step that needs to be taken to develop drugs for their treatment. Animal models are being increasingly used to expand the knowledge and understanding on neurodegenerative diseases. Marine worms, known as polychaetes (phylum Annelida), which are abundantly and frequently found in benthic environments, possess a simple yet complete nervous system (including a true brain that is centralised and specialised) compared to other annelids. Hence, polychaetes can potentially be the next candidate for a nerve disease model. The ability to activate the entire nervous system regeneration (NSR) is among the remarkable features of many polychaetes species. However, the information on NSR in polychaetes and how it can potentially model neurodegenerative diseases in humans is still lacking. By exploring such studies, we may eventually be able to circumvent the developmental constraints that limit NSR in the human nervous system. This article is intended to briefly review responsible mechanisms and signalling pathways of NSR in marine polychaetes and to make a comparison with other established models of neurodegenerative disease.

Penicilazaphilone C, a new azaphilone, induces apoptosis by blocking the Notch signalling pathway in gastric cancer

Objective: To explore the molecular mechanisms of the anticancer activities of penicilazaphilone C against gastric cancer. Methods: In vitro effects of penicilazaphilone C on cell growth, proliferation, and apoptosis were evaluated by MTT, BrdU, MTS, colony formation assays, Hoechst 33258 staining, and flow cytometry. Related proteins were examined by Western blotting assays. The expression of Notch receptor was analyzed using real-time PCR. In vivo antitumor activities of penicilazaphilone C were observed in nude mice. Results: Compared to the controls, penicilazaphilone C suppressed cell proliferation and promoted apoptosis in MGC-803 and SGC-7901 cells. The Notch/PTEN/AKT axis was involved in the activating penicilazaphilone C-induced apoptosis. Penicilazaphilone C decreased levels of Notch, NICD, phospho-PTEN and phospho- AKT compared to controls. The penicilazaphilone C-induced inhibition of Notch-related protein expression levels and the resulting apoptosis could be reversed by overexpression of Notch1 or/and Notch2. Moreover, penicilazaphilone C inhibited tumor growth in mice bearing tumours derived from MGC-803 and SGC-7901 cells, respectively. Conclusions: Penicilazaphilone C can induce the apoptosis by suppressing the activation of the proteolytic cleavage of the Notch receptor and subsequently blocking the PTEN/AKT signaling axis in gastric cancer cells. Thus, penicilazaphilone C is a potential alternative agent for the treatment of gastric cancer.

S100 proteins: An emerging cynosure in pregnancy & adverse reproductive outcome

S100 proteins are calcium (Ca2+)-binding proteins and these have an important function in progression, manifestation and therapeutic aspects of various inflammatory, metabolic and neurodegenerative disorders. Based on their involvement in intracellular or extracellular regulatory effects, S100 proteins are classified into three subgroups: one subgroup is specialized in exerting only intracellular effects, other performs both intracellular and extracellular functions and the third subgroup members only display extracellular regulatory effects. S100 proteins are expressed particularly in vertebrates and have cell-specific expression. Functionally, S100 proteins act through their surface receptors and regulate cell functions in autocrine or paracrine mode. Receptor for advanced glycation end products (RAGEs) and toll-like receptor 4 are the main surface receptors. S100 proteins participate in the regulation of cellular differentiation, proliferation, apoptosis and inflammation along with Ca2+ homeostasis, energy metabolism and cellular migration, and perform the respective functions through their interaction with transcription factors, nucleic acids, enzymes, receptors, cytoskeleton system, etc. Currently, their role in adverse pregnancy outcomes and compromised reproductive health is being explored. These proteins are present in amniotic fluid, endometrium tissue and foetal brain; therefore, it is quite likely that alterations in the expression levels of S100 family members will be affecting the particular function they are involved in and ultimately affecting the pregnancy in adverse manner. The current review discusses about an association of S100 proteins in pregnancy disorders such as endometriosis, intrauterine growth retardation and miscarriage.

Dengue virus stimulation of dendritic cells induces phosphorylation- and proteolysis-dependent signalling

Aims@#Dengue virus (DENV) infection is commonly observed in countries with tropical climates and remains a significant health hazard. No real cure has been established for the infection to date. @*Methodology and results@#To better understand the very early molecular events during the initial infection process, we exposed primary dendritic cells with Dengue virus and analysed proteins with increased phosphorylation signatures in the first 10 min using phospho-protein enrichment and tandem mass spectrometry analysis. Upon initial viral interaction, strong phosphorylation was observed for Endoplasmin, BiP, BID, Dok-2, GEF-H1 and Calpain-2. Reduced phosphorylation was noted for Importin-5, ERp72 and Rho-GDI. Knockdown of Calpain-2, a protease activated by calcium flux, reduced DENV infection rates of primary dendritic cells as measured by focus-forming units (FFUs). @*Conclusion, significance and impact of study@#We conclude that Calpain-2, BID, Importin 5 and ATP/GTPases are all active along the apoptosis pathway axis, indicating that dendritic cells commit to early signalling steps of cell death upon initial viral contact.

Transcriptomic analysis identifies differentially expressed genes (DEGs) associated with bolting and flowering in Saposhnikovia divaricata / 中国天然药物

Saposhnikovia divaricata is a valuable Chinese medicinal herb; the transformation from vegetative growth to reproductive growth may lead to the decrease of its pharmacological activities. Therefore, the study of bolting and flowering for Saposhnikovia divaricata is warranted. The present study aimed to reveal differentially expressed genes (DEGs) and regularity of expression during the bolting and flowering process, and the results of this study might provide a theoretical foundation for the suppression of early bolting for future research and practical application. Three sample groups, early flowering, flower bud differentiation, and late flowering (groups A, B, and C, respectively) were selected. Transcriptomic analysis identified 67, 010 annotated unigenes, among which 50, 165 were differentially expressed including 16, 108 in A vs B, and 17, 459 in B vs C, respectively. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway functional classification analysis were performed on these differentially expressed genes, and five important pathways were significantly impacted (P ≤ 0.01): plant circadian rhythm, other glycan degradation, oxidative phosphorylation, plant hormone signal transduction, and starch and sucrose metabolism. Plant hormone signal transduction might play an important role in the bolting and flowering process. The differentially expressed indole-3-acetic acid (IAA) gene showed significant down-regulation during bolting and flowering, while the transport inhibitor response 1 (TIR1) gene showed no significant change during the bolting process. The expression of flowering related genes FLC, LYF, and AP1 also showed a greater difference at different development stages. In conclusion, we speculate that the decrease in auxin concentration is not caused by the degrading effect of TIR1 but by an alternative mechanism.

IL-6 promotes MDSCs infiltration and immunosuppression in breast cancer by inducing SOCS3 deficiency / 中国肿瘤生物治疗杂志

@# Objective: To investigate the immunosuppressive effect and underlying molecular mechanisms of Myeloid-derived suppressor cells (MDSCs) on T cells activity through IL-6activatingSTAT3/IDO signaling pathway. Methods: Twenty pairs of cancer tissues and the corresponding adjacent normal tissues from breast cancer patients treated at Tianjin Medical University Cancer Institute and Hospital from November 2015 to February 2016 were collected for this study; in the meanwhile, peripheral blood samples from 40 healthy donorswere also collected. CD33+ cells in tumor tissues and CD33+ CD14 + cells in peripheral blood of helthy donors were sorted out with MicroBeads technology. CD33+ cells were in vitro co-cultured with breast cancer cell line MDA-MB-231 to induce MDSCs. Flow cytometry was used to detect the proportion of CD45+ CD13+CD33+CD14-CD15- MDSCs.Western Blotting was used to detect the expression ofSOCS1，SOCS3, JAK1, JAK2, TYK2, STAT1, STAT3 and their phosphorylation levels. qRT-PCR was used to detect the expression of IL-6 and SOCS1-3. CCK8 was used to detect the T cell proliferation. Annexin V staining was used to detect T cell apoptosis. ELISA was used to detect IL-10 and IFN-γ secreted by T cells. Results: There were MDSCs infiltration in all 20 cases of breast cancer tissues for different levels (15.3%~58.1%), with a mean level of (29.82± 11.46%); the infiltration of IL-6high group was significantly higher than that of the IL-6low group [（13.75±3.44） % vs（4.31±1.50） %， P< 0.05], indicating that IL-6 expression was positively correlated with MDSCs infiltration (R2=0.4399, P<0.01). In vitro experiments showed that tumor-derived IL-6 significantly promoted the generation and immunosuppressive activity of MDSCs (P<0.05), which could be reversed by the blocking of IL-6. In the meanwhile, the expression of SOCS3 in MDSCs that induced in vitro was absent, which can be inhibited by blocking IL-6 (P<0.05). Conclusion: The study has demonstrated that tumor-derived IL-6 stimulates the continuous activation of the JAK/STAT signaling pathway and the absence of SOCS3 expression in MDSCs, thereby promoting the infiltration, generation and immunological activity of MDSCs. Therefore, IL-6 signaling pathway can be used as therapeutic target to weaken MDSCs generation and reverse MDSCs activity.

The tankyrase inhibitor G007-LK inhibits small intestine LGR5+ stem cell proliferation without altering tissue morphology

Abstract Background The WNT pathway regulates intestinal stem cells and is frequently disrupted in intestinal adenomas. The pathway contains several potential biotargets for interference, including the poly-ADP ribosyltransferase enzymes tankyrase1 and 2. LGR5 is a known WNT pathway target gene and marker of intestinal stem cells. The LGR5+ stem cells are located in the crypt base and capable of regenerating all intestinal epithelial cell lineages. Results We treated Lgr5-EGFP-Ires-CreERT2;R26R-Confetti mice with the tankyrase inhibitor G007-LK for up to 3 weeks to assess the effect on duodenal stem cell homeostasis and on the integrity of intestinal epithelium. At the administered doses, G007-LK treatment inhibited WNT signalling in LGR5+ stem cells and reduced the number and distribution of cells traced from duodenal LGR5+ stem cells. However, the gross morphology of the duodenum remained unaltered and G007-LK-treated mice showed no signs of weight loss or any other visible morphological changes. The inhibitory effect on LGR5+ stem cell proliferation was reversible. Conclusion We show that the tankyrase inhibitor G007-LK is well tolerated by the mice, although proliferation of the LGR5+ intestinal stem cells was inhibited. Our observations suggest the presence of a tankyrase inhibitor-resistant cell population in the duodenum, able to rescue tissue integrity in the presence of G007-LK-mediated inhibition of the WNT signalling dependent LGR5+ intestinal epithelial stem cells.

The active substance of Rostellularia procunbens and its mechanism in inhibiting nephritis cell proliferation / 中成药

AIM To identify the active anti-chronic nephrotic substance of Rostellularia procunbens (L.) Nees,and to study its mechanism.METHODS Rat glomerular mesangial cells (HBZY-1) were developed into nephrotic cell models by LPS.The activities of extract of petroleum ether,ethyl acetate,n-butanol and water were screened by MTT and ELISA kit,after which isolation and purification of the various compounds were achieved,and their effects on the expression of TLR4/NF-κB pathway were determined by Western blot.RESULTS Both extracts of petroleum ether and ethyl acetate exhibited anti-nephrotic activity,and Justicidin A was determined to be the active compound inhibiting both the proliferation of mesangial cells and the release of cytokines to some extent.CONCLUSION Rostellularia procunbens (L.) Nees may inhibit the expression of inflammatory proteins through TLR4/NF-κB signalling pathway to prevent chronic nephritis.