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Childhood moyamoya disease (MMD) can lead to progressive and irreversible neurological impairment. Early age at onset is likely associated with a worst prognosis of the disease. The study aims to summarize the clinical characteristics of childhood MMD for supporting the diagnosis and treatment of early MMD.A retrospective study was conducted on children aged zero to 16 years who were diagnosed with MMD in the Department of Neurology and neurosurgery of our hospital from October 2016 to April 2020. The clinical characteristics of children with MMD were summarized for analysis, and the distribution of sex and initial attack type among different age groups was determined by data comparison.The study surveyed 114 children (male to female sex ratio of 1:1.07) with MMD, and 6.1% of them had family history. The mean age of onset was 7.15 ± 3.30 years, and the peak age of onset was five to eight years. The most common initial attack type was transient ischemic attack (TIA) (62 cases, 54.4%) with limb weakness. The incidence of the initial attack type in the three age groups was varied (P 0.05). The result of overall prognosis was good in 86 cases (89.6%).In this study, MMD cases were mainly ischemic type and TIA was the most common initial attack type. Infant group was more prone to have cerebral infarction, whereas preschool and school-age groups tended to have TIA. The treatments and prognosis of the studied MMD cases were achieved with good outcomes.

Natural killer (NK) cell therapies show potential for tumor treatment but were immunologically resisted by the overexpressed immunosuppressing tumor-cell-surface glycans. To reverse this glycan-mediated immunosuppression, the surface NK-inhibitory glycan expressions need to be downregulated and NK-activating glycan levels should be elevated synchronously with optimal efficiency. Here, we design a core-shell membrane-fusogenic liposome (MFL) to simultaneously achieve the physical modification of NK-activating glycans and biological inhibition of immunosuppressing glycans on the tumor cell surface via a membrane-fusion manner. Loaded into a tumor-microenvironment-triggered-degradable thermosensitive hydrogel, MFLs could be conveniently injected and controllably released into local tumor. Through fusion with tumor cell membrane, the released MFLs could simultaneously deliver sialyltransferase-inhibitor-loaded core into cytoplasm, and anchor NK-activating-glycan-modified shell onto tumor surface. This spatially-differential distribution of core and shell in one cell ensures the effective inhibition of intracellular sialyltransferase to downregulate immunosuppressing sialic acid, and direct presentation of NK-activating Lewis X trisaccharide (LeX) on tumor surface simultaneously. Consequentially, the sialic acid-caused immunosuppression of tumor surface was reprogrammed to be LeX-induced NK activation, resulting in sensitive susceptibility to NK-cell-mediated recognition and lysis for improved tumor elimination. This MFL provides a novel platform for multiplex cell engineering and personalized regulation of intercellular interactions for enhanced cancer immunotherapy. This article is protected by copyright. All rights reserved.

This paper describes a simple, low-cost, and fast method for synthesizing high-purity porous vaterite calcium carbonate (CaCO3) without any additives using steamed ammonia liquid waste and (NH4)2CO3. The obtained vaterite samples were characterized using various techniques such as powder X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), and laser particle size analysis (LPSA). The XRD and FTIR analyses indicate that the prepared product is high-purity vaterite CaCO3. The SEM analysis shows obtained micro/nanosized spherical and porous vaterites. The effects of stirring mode and stirring speed on the morphology, particle size, and stability of vaterite were studied systematically. The stirring speed and stirring mode were found to play an important role in the crystallization of vaterite CaCO3 with different purities, morphologies, particle sizes, and particle size distributions. Based on the analysis of vaterite CaCO3 particles obtained under different stirring modes and stirring speeds, a possible mechanism for obtaining smaller vaterite CaCO3 particles at high stirring speed is proposed. The possible mechanism underlying the properties of ammonia-distilled waste liquid during vaterite formation is also discussed.

The effect of underwater radiated noise (URN) pollution (produced by merchant ships) on marine ecology has become a topic of extreme concern for both the academic community and the general public. This paper summarises some research results and modelling about shipping noise published over several decades, which comprises the research significance of low-frequency ambient noise and shipping noise, shipping noise source levels (SL), empirical models and the measurement standards of shipping noise. In short, we try to present an overall outline of shipping noise and ocean ambient noise for related research.

Background Plank exercise (PE) is a whole-body isometric muscle training which is beneficial for physical health. However, none of the previous studies investigated the responses within a typical isometric muscle training or PE protocol consisting of multiple sets. The application of PE was restricted for the understudied metabolic and cardiovascular responses, especially for the patients with cardiovascular diseases. This study is to alleviate the safety concerns of PE by investigating the PE-induced metabolic and cardiovascular responses. Methods Eleven male recreational-level college students completed a baseline cardiopulmonary exercise test, continuous PE (CPE) and intermittent PE (IPE). Ratio of maximal oxygen uptake per kilogram of body mass (%VO2max/kg), ratio of maximal heart rate (%HRmax), and respiratory exchange ratio (RER) were continuously measured during PEs and divided into seven equal timepoints. Blood pressure (BP) was measured every minute during, before, and after PEs. A mixed-model repeated measures ANOVA was used to examine the interaction effect of exercise × phase. Results The %VO2max/kg (F6,69=11.25, P max (F6,65=7.74, P 6,69=11.56, P 2,26=8.42, P = 0.002; diastolic BP, F2,24=22.63, P 2max/kg (33.5 [2.2] vs. 27.7 [1.9], P = 0.043) and %HRmax (63.2 [3.9] vs. 53.3 [2.1], P = 0.019) increased more significantly from the 40% duration of CPE. Systolic BP increased by larger magnitudes during CPE than IPE (154.2 [3.8] vs. 142.3 [4.8] mmHg, P = 0.002). RERs were over 1 during PEs without cardiovascular and metabolic variables over the anaerobic threshold. Conclusion Energy was mainly supplied by anaerobic metabolism during PEs. CPE may be preferable for trainees aiming at anaerobic capacity enhancement. IPEs may be preferable to CPEs for youth patients with mild and borderline cardiovascular diseases due to their lower metabolic and cardiovascular responses.

Membrane fusion, a fundamental biological process of the fusion of the membrane composition between cells, is vital for cell-cell communication and cargo transport between living cells. This fusion interaction achieves the transportation of the inner content to the cellular cytosol as well as the simultaneous blending of foreign substances with the cell membrane. Inspired by this biological process, emerging membrane-fusogenic particles have been developed, opening a new area for bioengineering and biomedical applications. Especially, membrane-fusion-mediated transfer of inner cargoes can bypass endosomal entrapment to maximize the transportation efficiency, emerging as a unique cytoplasmic delivery platform distinct from those depending on conventional endocytosis-based pathways. In addition, the membrane fusion enables cell surface modification through lipid diffusion and mixing, providing a tool for direct cell membrane engineering. In this review, we focus on the development of membrane-fusogenic particles and their up-to-date progress. We briefly introduce the concept of membrane fusion, elaborate inspiring strategies of membrane-fusogenic particles, and highlight the recent advances and the promising applications of membrane-fusogenic particles as a next-generation bioengineering tool. In the end, we conclude with the present challenges and opportunities, providing insights in the future research of membrane-fusogenic particles.

Identifying high specificity and sensitivity biomarkers has always been the focus of research in the field of non-invasive cancer diagnosis. Exosomes are extracellular vesicles with a lipid bilayer membrane that can be released by all types of cells, which contain a variety of proteins, lipids, and a variety of non-coding RNAs. Increasing research has shown that the lipid bilayer can effectively protect the nucleic acid in exosomes. In cancers, tumor cell-derived exosomal circRNAs can act on target cells or organs through the transport of exosomes, and then participate in the regulation of tumor development and metastasis. Since exosomes exist in various body fluids and circRNAs in exosomes exhibit high stability, exosomal circRNAs have the potential as biomarkers for early and minimally invasive cancer diagnosis and prognosis judgment. In this review, we summarized circRNAs and their biological roles in cancers, with the emerging value biomarkers in cancer diagnosis, disease judgment, and prognosis observation. In addition, we briefly compared the advantages of exosomal circRNAs as biomarkers and the current obstacles in the exosome isolation technology, shed light to the future development of this technology.

To analyze the clinical and genetic characteristics of a child with clinical manifestations of hypoplasia, epilepsy and abnormal face.The clinical data of the child were collected. The peripheral blood samples of the patient and his parents were extracted for high-throughput sequencing, and Sanger sequencing verification and bioinformatics analysis were performed to detect suspected pathogenic variants.The clinical manifestations of the child were overall developmental backwardness, seizures, autism, and special facial appearance. High throughput sequencing showed that there was a heterozygous mutation of exon 11: c.1920_c.1927delCCTCTACC (p.Ser641Rfs*31) of the DYRK1A gene. The same variant was found in neither of her parents, suggesting that it has a denovo origin.The exon11: c.1920_c.1927delCCTCTACC (p.Ser641Rfs*31) mutation in DYRK1A gene was the genetic etiology of the case, which enriches the pathogenic gene spectrum of DYRK1A and provides the basis for clinical diagnosis and genetic counseling.

MicroRNAs regulate gene expression at the posttranscriptional level, and this process has been shown to be implicated in the pathological processes of temporal lobe epilepsy. At present, studies about the impact of microRNA-181a (miR-181a) on epilepsy have focused on hippocampal neurons, and the effect of miR-181a on other cells in the hippocampus remains poorly understood. Herein, we explored the role of miR-181a-5p in a lithium-pilocarpine model of epilepticus in immature rats. We found that the hippocampal expression level of miR-181a-5p was increased. Inhibition of miR-181a-5p protected the hippocampus against epilepsy, including hippocampal insults, neuronal apoptosis, astrocyte and microglia activation, neuroinflammation, oxidative stress, mitochondrial function, and cognitive dysfunction. Moreover, miR-181a-5p inhibition exerted a seizure-suppressing effect via SIRT1 upregulation. Overall, our findings reveal the potential role of the miR-181a-5p/SIRT1 pathway in the development of temporal lobe epilepsy, and this pathway may represent a novel target for ameliorating epilepsy and its sequelae.

Purpose Pediatric acute promyelocytic leukemia (APL) accounts for 10% of pediatric acute myelogenous leukemia (AML) case and is accompanied by a tendency to hemorrhage. miR-188-5p plays an important role in adult AML. Therefore, the purpose of this study was to explore the effects of miR-188-5p on cell proliferation and apoptosis and tumor growth, and its mechanism in pediatric APL patients. Materials and methods Survival-associated miRNAs or mRNAs from TCGA database associated with AML were identified via using the "survival R" package in R language. CCK8, clone formation, flow cytometry, RT-PCR, immunohistochemistry and Western blot assays were used to detect the viability, proliferation, apoptosis, cell cycle, and related gene expression in APL cell lines. The prognostic value of miR-188-5p was evaluated using a ROC curve. The tumorigenic ability of APL cell lines was determined using a nude mouse transplantation tumor experiment. Tumor cell apoptosis was determined by TUNEL assay in vivo. The target genes of miR-188-5p were predicted using the miRDB, miRTarBase, and TargetScan databases. A PPI network was constructed using STRING database and the hub gene was identified using the MCODE plug-in of the Cytoscape software. The DAVID database was used to perform GO and KEGG pathway enrichment analyses. A luciferase reporter assay was used to demonstrate the binding of miR-188-5p to CD2AP. Results miR-188-5p overexpression or CD2 associated protein (CD2AP) inhibition was significantly associated with poor survival in pediatric APL patients. Upregulation of miR-188-5p was identified in the blood of pediatric APL patients and cell lines. Increased expression of miR-188-5p also promoted the viability, proliferation, and cell cycle progression, and reduced the apoptosis of APL cells. Additionally, upregulation of miR-188-5p regulated the expressions of cyclinD1, p53, Bax, Bcl-2 and cleaved caspase-3. The area under the ROC curve (AUC) of miR-188-5p was 0.661. miR-188-5p overexpression increased the tumorigenic ability of APL and Ki67 expression, and reduced cell apoptosis in vivo. CD2AP was identified as the only overlapping gene from the list of miR-188-5p target genes and survival-related mRNAs of the TCGA database. It was mainly enriched in the "biological process (BP)" and "cellular component (CC)" terms, and was downregulated in the blood of pediatric APL patients and cell lines. The luciferase reporter, RT-PCR, and Western blot assays demonstrated that the binding of miR-188-5p to CD2AP. CD2AP inhibition promoted the proliferation and inhibited the apoptosis of APL cells. Rescue experiments showed that inhibition of miR-188-5p inhibited cell proliferation, activated the PI3K/AKT/mTOR signaling pathway, induced G0/G1 phase arrest, regulated gene expression, and promoted cell apoptosis, which were reversed by CD2AP inhibition. Conclusion miR-188-5p, an oncogene, promoted tumor growth and progression of pediatric APL in vitro and in vivo via targeting CD2AP and activating the PI3K/AKT/mTOR signaling pathway.

Introduction Berberine has been reported to inhibit cancer cell growth by apoptosis induction and exhibits a protective role against cancer progression. The current study aims to investigate the effects of berberine on acute lymphoblastic leukemia (ALL) and the mechanism beyond apoptosis. Methods Cell viability was determined in ALL cell lines EU-6 and SKW-3 using trypan blue staining. Cell autophagy was determined by immunofluorescence and Western blot. ALL xenograft mice were established to investigate the anti-tumor effects of BBR. The molecular mechanism was explored in ALL cell lines using siRNA and signaling inhibitors. Results Herein, we show that berberine treatment significantly inhibits ALL cell viability and promotes cell death by inducing autophagy in a dose-dependent manner. Moreover, berberine significantly alleviates the aggressive pathological condition in ALL xenograft mice. Mechanistic studies exhibit that berberine induces autophagic death in ALL cells by inactivating AKT/mTORC1 signaling. Chemically targeting AKT/mTORC1 signaling controls berberine-induced cell autophagy in vitro, and blockade of autophagic process blunts berberine-alleviated pathological condition in vivo. Discussion In conclusion, our study reveals that berberine could induce ALL cell autophagic death by inactivating AKT/mTORC1 signaling that could be used to develop small molecule drug for ALL treatment.

CRISPR Nanotheranostics In article number 2102051, Yeh‐Hsing Lao, Qi Zhang, Yu Tao, Mingqiang Li, and co‐workers overview the recent advances of CRISPR nanotheranostics in tacking viral hepatitis and hepatocellular carcinoma. The nanoscale CRISPR/Cas systems could act as smart enzymatic robots to target the disease‐associated cells and nucleic acid signatures and may be potentially added to the armamentarium for efficiently addressing these challenging liver diseases. [Image: see text]

A colloidal nanocrystal cluster (CNC) is a hierarchical nanostructure formed by clustering several nanocrystals into one nano-ensemble, which may exhibit unique optical or catalytic properties different from individual nanocrystals owing to the mutual interactions among neighboring component nanocrystals. However, there is still no universal synthetic route that could be applicable to diverse material compositions with precisely controlled hierarchical structures (i.e., nanocrystal number density, component nanocrystal size, and overall diameter of the CNC) up to now. Herein, a general and novel synthetic strategy was reported for crafting a wide range of inorganic CNCs (i.e., noble metal, semiconductor, and metal oxide) via utilizing amphiphilic star-like poly(4-vinylpyridine)-block-polystyrene diblock copolymers as nanoreactors prepared by sequential atom transfer radical polymerization. The hierarchical structure of rationally designed CNCs could be readily tailored by varying the P4VP molecular weight of star-like nanoreactors and the parameter optimization during the CNC preparation process, which was inaccessible by conventional synthetic methods.

Liver disease, particularly viral hepatitis and hepatocellular carcinoma (HCC), is a global healthcare burden and leads to more than 2 million deaths per year worldwide. Despite some success in diagnosis and vaccine development, there are still unmet needs to improve diagnostics and therapeutics for viral hepatitis and HCC. The emerging clustered regularly interspaced short palindromic repeat/associated proteins (CRISPR/Cas) technology may open up a unique avenue to tackle these two diseases at the genetic level in a precise manner. Especially, liver is a more accessible organ over others from the delivery point of view, and many advanced strategies applied for nanotheranostics can be adapted in CRISPR‐mediated diagnostics or liver gene editing. In this review, the focus is on these two aspects of viral hepatitis and HCC applications. An overview on CRISPR editor development and current progress in clinical trials is first given, followed by highlighting the recent advances integrating the merits of gene editing and nanotheranostics. The promising systems that are used in other applications but may hold potentials in liver gene editing are also discussed. This review concludes with the perspectives on rationally designing the next‐generation CRISPR approaches and improving the editing performance., Herein, the clustered regularly interspaced short palindromic repeat (CRISPR) nanotheranostics developed for viral hepatitis and hepatocellular carcinoma are reviewed, and rational design considerations for the next‐generation nanomedicine approaches to advance liver gene editing are discussed, highlighting their significances in clinical applications.

Elderly with comorbidities have shown a higher rate of fatal outcomes when suffering coronavirus disease 2019 (COVID-19). However, a delineation of clinical significances of hematologic indices and underlying comorbidities in the progression and outcome of COVID-19 remains undefined. Six hundred two COVID-19 patients with established clinical outcomes (discharged or deceased) from Hankou Hospital of Wuhan, China between January 14, 2020 and February 29, 2020 were retrospectively analyzed. Of the 602 patients with COVID-19, 539 were discharged and 63 died in the hospital. The deceased group showed higher leukocyte and neutrophil counts but lower lymphocyte and platelet counts. Longer activated partial thromboplastin time (APTT) and prothrombin time (PT), as well as higher D-dimer and C-reactive protein levels, were found in non-survivors. Our observations suggest that these parameters could serve as potential predictors for the fatal outcome and in the discharged group. A higher neutrophil count and D-dimer level but lower lymphocyte were associated with a longer duration of hospitalization. A multivariable Cox regression analysis showed that higher neutrophil count, prolonged PT, and low lymphocyte count were risk factors for patients with COVID-19. Also, we found an association of lower lymphocyte count and higher C-reactive protein levels with the elderly group and those with cardiovascular-related comorbidities. The significantly different hematologic profiles between survivors and non-survivors support that distinct hematologic signatures in COVID-19 patients will dictate different outcomes as a prognostic marker for recovery or fatality. Lymphopenia and aggressive inflammatory response might be major causes for fatal outcomes in the elderly male and especially those with cardiovascular-related comorbidities.

Objectives: Chinese herb medicine (CHM) is one of the most popular complementary and alternative therapies, which has been widely used to treat Refractory Mycoplasma Pneumoniae Pneumonia (RMPP). However, the effect and safety of CHM remain controversial. Hence, we conducted this meta-analysis to evaluate whether CHM combination therapy could bring benefits to children and adolescents with RMPP.Methods: Seven databases were used for data searching through November 11, 2020 following the PRISMA checklist generally. Review Manager 5.3, Trial sequential analysis 0.9.5.10 Beta software and Stata16.0 were applied to perform data analyses. Mean difference or risk ratio was adopted to express the results, where a 95% confidence interval (CI) was applied.Results: In general, this research enrolled 17 trials with 1,451 participants. The overall pooled results indicated that CHM was beneficial for children and adolescents with RMPP by improving the clinical efficacy rate [RR = 1.20, 95% CI (1.15, 1.25), p < 0.00001], shortening antipyretic time [MD = −2.60, 95% CI (−3.06, −2.13), p < 0.00001], cough disappearance time [MD = −2.77, 95% CI (−3.12, −2.42), p < 0.00001], lung rale disappearance time [MD = −2.65, 95% CI (−3.15, −2.15), p < 0.00001], lung X-ray infiltrates disappearance time [MD = −2.75, 95% CI (−3.33, −2.17), p < 0.00001], reducing TNF-α level [MD = −5.49, 95% CI (−7.21, −3.77), p < 0.00001]. Moreover, subgroup results suggested that removing heat-phlegm and toxicity therapy had more advantages in shortening antipyretic time, cough disappearance time, lung X-ray infiltrates disappearance time and reducing TNF-α level. Meanwhile promoting blood circulation therapy seemed to be better at relieving lung rale. However, regarding adverse events, the two groups displayed no statistical difference [RR = 0.97, 95% CI (0.60, 1.57), p = 0.91].Conclusion: Despite of the apparently positive results in relieving clinical symptoms, physical signs and reducing inflammation, it is premature to confirm the efficacy of CHM in treating RMPP because of the limitation of quality and the number of the included studies. More large-scale, double-blind, well-designed, randomized controlled trials are needed in future research.

We reported the synthesis of a well-defined hollow polymer nanoparticle derived from star-shaped unimolecular micelles. β-Cyclodextrin was first applied as an efficient macroinitiator to prepare a star-shaped PCL via ring-opening polymerization (ROP). Then, the star-shaped PCL was modified to be a macro-RAFT agent for photoinduced electron/energy transfer-reversible addition-fragmentation chain transfer (PET-RAFT) polymerization of S-Cl monomers. The prepared unimolecular micelles can be photocross-linked under UV irradiation after a simple nucleophilic substitution reaction, which made -Cl groups to be -N3 groups. After the selective removal of the PCL core, hollow polymer nanoparticles were achieved and exhibited to be a general nanoreactor strategy for the fabrication of nanocrystals with well-controlled architectures. Compared with unimolecular micelle templates, the nanocrystals prepared by hollow templates are absolutely pure as no polymer chains are embedded in the inorganic nanocrystals. In addition, by changing the concentration of the precursor, the structure of the nanocrystal can be changed from a normal spherical structure to a hollow structure.

Lead (Pb) isotopes and strontium (Sr) isotopes are good combined multiple geochemical tracers to differentiate lead pollution in soil and groundwater due to the characteristic “fingerprints” of Pb isotopes and fractionation of Sr isotopes. Soil, surface water, groundwater, and river sediment samples were investigated for Pb and Sr content and the isotopic ratio was used to determine the potential sources of Pb in soil and groundwater of the Hun River alluvial fan, Liaoning, China. Lead from anthropogenic sources was of high solubility and could translocate, and it was relatively stable after deposition in the soil. Therefore, lead speciation and the isotopic ratio of the acid extraction fraction and the residual fraction in the soil and river sediments were also measured to obtain more information regarding the anthropogenic pollution. The results indicate that most Pb in the soil was of geogenic origin, in particular, ore chemical weathering along the Hun River, while the topsoil was significantly affected by leaded gasoline and coal combustion, sewage irrigation, and other human activities. The geogenic origin of Pb in groundwater was from the carbonate rock aquifer and ore upstream. Shallow groundwater was affected by contaminated surface water through the interaction of groundwater and rivers. The influence of vehicle exhausts and coal combustion on groundwater from municipal wells can be ignored.

Background: Thrombospondin 1 (TSP-1) is an extracellular matrix protein that interacts with a wide array of ligands including cell receptors, growth factors, cytokines, and proteases to regulate various physiological and pathological processes. TSP-1 induces apoptosis of endothelial and cancer cells via its receptor CD36. This study was to investigate the effect of TSP-1 on apoptosis of human megakaryocytic leukemia cell line Meg-01 and its possible mechanism. Methods: The expression of CD36 antigen in Meg-01 cells was detected by flow cytometry and immunocytochemistry. Meg-01 cells were cultured for 48 hours with TSP-1 and CD36 antibody FA6-152 at different concentrations, to investigate the effect of TSP-1 on the growth of megakaryocytes. The early apoptosis and the activity of the apoptotic protease caspase-3 were detected by flow cytometry. Bone marrow cells of mice were cultured for CFU-MK and stained with acetylcholine to detect the differentiation of megakaryocytes. Results: CD36 antigen was detected on the surface of Meg-01 cells by flow cytometry and immunocytochemistry. TSP-1 (5 μg/mL) inhibited the proliferation of Meg-01 cells, but not M-07e cells (CD36 -). After the addition of CD36 antibody FA6-152 (5, 10 and 25 μg/mL), the inhibitory effect of TSP-1 was significantly reduced. TSP-1 (2.5, 5 and 7.5 μg/mL) exerted a pro-apoptotic effect by increasing the expression of Annexin V (P Conclusion: TSP-1 could inhibit the proliferation of Megakaryocyte cell line Meg-01 cells, and induce it`s apoptosis. TSP-1 may induce apoptosis of Meg-01 cells via CD36 or caspase-3, which provides a potential new drug choice for clinical treatment of megakaryocytic leukemia. Disclosures No relevant conflicts of interest to declare.

Background: Angelica Polysaccharide (APS) is from the root of Radix Angelicae Sinensis (Danggui). Danggui has been used for centuries to treat blood-deficiency related diseases. The hematopoietic effect of Danggui may be related to its constituent, polysaccharide. The effects of angelica polysaccharide on cryopreservation of platelets and megakaryocytes have not been well studied. This study focused on anti-apoptotic effect of APS and TPO on cryopreservation of platelets and megakaryocytes and provided new methods for prolonging the preservation time of platelets in vitro. Methods: The expression of platelet membrane glycoprotein CD41 and CD61, as well as the platelet apoptotic rate, Caspase 3 expression and mitochondrial membrane potential (MMP) were detected by flow cytometry; the anti-apoptotic mechanism of APS by PI3K /AKT signaling pathway was analyzed by Western blot assay. CFU assays were used to determine the effects of APS on megakaryocytic progenitor cells. Analyses of Annexin V, Caspase-3, and Mitochondrial Membrane Potential were conducted in megakaryocytic cell line M-07e. The effects of APS on cells treated with Ly294002, PI3K inhibitor and the effect of APS on the p-AKT were also studied. Results: The platelets were divided into 4 group: control group (4 ℃ stored platelets), APS group (APS-treated platelets stored at 4 ℃), LY294002 group (LY294002-treated platelets stored at 4 ℃) and LY294002+APS group (LY294002+APS treated platelets stored at 4 ℃). The apoptotic rate of platelets in LY294002 group was obviously increased. Compared with control group, the expression of CD41 and CD61 gradually decreased along with the enhancement of LY294002 concentrations (r=-0.953). The apoptotic rate of platelets in LY294002 group was enhanced significantly (P Conclusion: APS, like TPO, has an anti-apoptotic effect on the cryopreserved platelets and megakaryocytes through activating PI3K/AKT, decreasing the expression of Caspase-3 and inhibiting the reduction of MMP. Disclosures No relevant conflicts of interest to declare.

Background: Essential Thrombocythemia (ET) is characterized by persistently elevated platelet counts in the context of a normal red cell mass. However, the molecular mechanism of ET is still under investigation. Our previous studies demonstrated the presence of functional PDGF receptors (PDGFR) on megakaryocytes and their ability to mediate hematopoiesis and megakaryopoiesis (Ye et al, Haematologica, 2010). The role of PDGF-BB on megakaryocytic progenitors (CD41+, CD34+ cells) and its mechanisms on ET will be further studied in this project. Methods: ELISA, CFU assay, immunofluorescence microscope, flow cytometry and NOD/SCID mice were used in this study. Results: Bone marrow plasma levels of PDGF-BB in ET patients (n=18) and normal control (n=10) were tested and found an increased PDGF-BB levels in ET patients (2071.2±124.8 pg/ml), compared with normal control (1382.5±128.3pg/ml) (P=0.002). In vitro experiment, PDGF-BB promoted the ex vivo expansion of human hematopoietic stem (CD34 +) and progenitor (CD41 + CD61 +) cells. More significantly, PDGF enhanced the engraftment of human CD45 + cells and their myeloid subsets (CD33 +, CD14 + cells) in NOD/SCID mice. PDGF-BB stimulated megakaryopoiesis via PDGFR and its signalling. It also showed a direct stimulatory effect of PDGF-BB on c-Fos, GATA-1 and NF-E2 expressions in megakaryocytes. We speculate that these transcription factors might be involved in the signal transduction of PDGF-BB on the regulation of megakaryopoiesis. PDGF-BB also enhanced platelet recovery in mice model with radiation-induced thrombocytopenia. Studies showed that PDGF, like TPO, significantly promoted platelet recovery and the formation of CFU-MK in this irradiated-mouse. An increased number of hematopoietic stem/progenitor cells and a reduction of apoptosis were found in the bone marrow histology sections. In the CHRF apoptotic model, PDGF-BB had a similar anti-apoptotic effect as TPO on megakaryocytes. We also demonstrated that PDGF-BB activated the p -Akt , p-Jak2 and p-Stat3 expression, while addition of imatinib mesylate reduced p-Akt, p-Jak2 and p-Stat3 expression in CHRF cells. Conclusion: Our findings suggested that PDGF-BB is likely to be mediated via PDGF receptors with subsequent activation of the Akt and Jak2/ Stat3 pathways in megakaryopoiesis. These studies provide a possible explanation that PDGF-BB and its signaling may be involved in the molecular mechanism of essential thrombocythemia. Disclosures No relevant conflicts of interest to declare.

Introduction Berberine has been reported to inhibit cancer cell growth by apoptosis induction and exhibits a protective role against cancer progression. The current study aims to investigate the effects of berberine on acute lymphoblastic leukemia (ALL) and the mechanism beyond apoptosis. Methods Cell viability was determined in ALL cell lines EU-6 and SKW-3 using trypan blue staining. Cell autophagy was determined by immunofluorescence and Western blot. ALL xenograft mice were established to investigate the anti-tumor effects of BBR. The molecular mechanism was explored in ALL cell lines using siRNA and signaling inhibitors. Results Herein, we show that berberine treatment significantly inhibits ALL cell viability and promotes cell death by inducing autophagy in a dose-dependent manner. Moreover, berberine significantly alleviates the aggressive pathological condition in ALL xenograft mice. Mechanistic studies exhibit that berberine induces autophagic death in ALL cells by inactivating AKT/mTORC1 signaling. Chemically targeting AKT/mTORC1 signaling controls berberine-induced cell autophagy in vitro, and blockade of autophagic process blunts berberine-alleviated pathological condition in vivo. Discussion In conclusion, our study reveals that berberine could induce ALL cell autophagic death by inactivating AKT/mTORC1 signaling that could be used to develop small molecule drug for ALL treatment.

Multi-drug resistant infections caused by Gram-negative bacteria have become one of the most important reasons for the failure of clinical anti-infective treatment. Siderophore-antibiotic conjugates, which were designed based on a "Trojan horse" strategy wherein features enabled active uptake to bypass the Gram-negative cell wall, have been expected to be a weapon for anti-infective treatment in the clinic. Herein, we review antibiotic drug design strategies based on mimics of nature siderophores reported in recent years, we also focus our attention on the relationship between the type of linker and the corresponding antibacterial activity.

To detect potential mutation in a Chinese pedigree affected with split hand/split foot malformation (SHFM).The patients were screened for genome-wide copy number variations with single nucleotide polymorphism (SNP) microarray. Copy number variations were verified by real-time fluorescence quantitative PCR.There were 3 SHFM patients from three generations, which conformed to an autosomal dominant inheritance. SNP microarray assay revealed that all patients have carried a 0.34 Mb duplication in 10q24.31-q24.32 (102 993 649-103 333 271) encompassing the BTRC and DPCD genes. The result was verified by real-time fluorescence quantitative PCR, confirming that the duplication has co-segregated with the SHFM phenotype in the pedigree.The 10q24.31-q24.32 duplication probably underlies the pathogenesis of SHFM in this pedigree. Tiny copy number variations can result in diseases featuring autosomal dominant inheritance.

Program death-1 (PD-1) is a co-inhibitory receptor inducibly expressed on activated T cells. PD-1 has been reported to be associated with the development of several autoimmune diseases including rheumatoid arthritis, but the precise cellular and molecular mechanisms have not been fully elucidated. To study the role of PD-1 in the pathogenesis of rheumatoid arthritis and the possible underlying mechanisms, we performed collagen-induced arthritis (CIA) in C57BL/6 mice. Here, we show that PD-1 deficiency leads to the development of severe CIA in mice. When analyzing T cells from CIA mice ex vivo, we noticed aberrant antigen-specific Th17 responses in mice lacking PD-1. This is possibly due to deregulated activation of PKC-θ and Akt. In support of this notion, treating Pdcd1 (-/-) mice with an inhibitor of PI3-kinase that is upstream of PKC-θ and Akt significantly suppressed the disease severity. Therefore, our data indicate that PD-1 dampens antigen-specific Th17 response, thus inhibiting the disease.

Astrocyte activation, associated with the release of pro-inflammatory cytokines interleukin 1-β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor alpha (TNF-α), is a hallmark of multiple brain diseases, including mesial temporal lobe epilepsy. In recent years, several microRNAs have emerged as important controllers of Toll-like receptor (TLR) signaling. In this study, we investigated the effect of miR-132, miR-146a, and miR-155 on myeloid-related protein-8 (MRP8) induced astrocyte-related inflammation. Using quantitative polymerase chain reaction (qPCR) and western blot, we found clear upregulation of TLR4 and downstream inflammatory cytokines, along with dysregulation of miR-132, miR-146a, and miR-155 in in vitro astrocytes after exposing them to different concentrations of MRP8. In addition, we focused on the effect of miR-132 on astrocyte-related inflammation induced by MRP8 via lentiviral infection then evaluated the expression of its possible target genes: acetylcholinesterase (AChE) and interleukin-1 receptor-associated kinase (IRAK4). Our results show that miR-132 is a negative feedback regulator of IL-1β and IL-6, but not TNF-α, by targeting IRAK4. Together, our findings demonstrate the novel role of TLR4-related microRNAs, especially miR-132, in the regulation of MRP8-induced astrocyte activation and highlight the importance of miR-132 in the modulation of innate immune response induced by endogenous ligands in neurological diseases.

Although the pathogenesis and epileptogenesis of mesial temporal lobe epilepsy (MTLE) have been studied for years, many questions remain. The ubiquitin-proteasome system (UPS) is one factor that might regulate ion channels, inflammation and neuron excitability. Nedd4-2 is an E3 ubiquitin ligase linked with ion channels and synaptic vesicle recycling. Here, we explore the role of the UPS and its E3 ligase Nedd4-2 in the pathogenesis of MTLE. Our western blot results revealed that ubiquitin and Nedd4-2 were expressed differentially in different stages of MTLE. Co-immunoprecipitation and double immunostaining results indicated that Nedd4-2 was the substrate protein of ubiquitin both in vivo and in vitro. Inhibition of the UPS aggravated the epileptogenesis of MTLE, causing early and frequent spontaneous seizures, more obvious neuron loss and aberrant mossy fiber sprouting. Inhibition of ubiquitin also enhanced the activation of Nedd4-2, and switched ion channel α-ENaC downstream. Our study is the first to report that the UPS participates in the pathogenesis of MTLE, inhibition of UPS could aggravate the epileptogenesis, and that Nedd4-2 is a critical E3 ligase involved in this process.

A comprehensive model for groundwater risk assessment was proposed by combining the hazard of contaminated sources (H), groundwater intrinsic vulnerability (V) and groundwater function value (F), and verified in Jilin City of northeast China. This model is characterized by integrating groundwater resource, ecology, and geological environment under the impact of climate change and human activities. The hazard of potential polluted sources was assessed by quantifying the properties and the potential infiltrating load of contaminants. The groundwater intrinsic vulnerability was evaluated by the DRASTIC model. The groundwater function value was assessed by multiplying seventeen indexes with their corresponding weightings. The groundwater pollution risk mapping of Jilin City was generated based on ArcGIS and validated by the level difference method between the risk classification and the pollution classification. The results showed that groundwater has a relatively low possibility of pollution because the area with less than, or equal to, the medium classification of risk accounted for 67.61% of the study area. The hazard harmfulness from the different contaminant sources played the most important role in determining the high groundwater pollution risk areas. Different influencing factors lead to relatively high pollution risk in specific areas. According to the validation of the level difference method, areas correctly identified by groundwater pollution risk mapping accounted for 95.81% of the study area, which is nearly twice as high as that of specific vulnerability mapping. The HVF model proved to be suitable for assessing groundwater pollution risk in Jilin City of northeast China. The groundwater pollution risk mapping can be applied for the effective protection and sustainable supply of groundwater.

Objective Status epilepticus (SE) is a common, life-threatening neurological emergency that confers a high degree of morbidity and mortality. Increasing evidence indicates that neuroinflammation plays a critical role in the pathogenesis of SE. MicroRNA-146a (miR-146a) has been reported to be an important posttranscriptional inflammation-associated microRNA. The aim of this study was to investigate the effect of miR-146a in SE and the mechanism by which it operates. Methods To study the effect of miR-146a in SE, we chose intracerebroventricular injection for rat at 21–28 days old, and made a lithium-pilocarpine-induced SE rat model. We assessed latency time and Lado grade by behavior observation. We performed qPCR, ELISA and western blot tests on rat hippocampus to measure the expression levels of miR-146a, IL-1β, TNF-α, TLR4 and NF-κB. Results In the miR-146a antagomir injection group, the latency to generalized convulsions was shorter, the duration and degree of seizures were more severe, the expression level of miR-146a was clearly decreased, and IL-1β, TNF-α, TLR4 and NF-κB were all significantly up-regulated. The opposite was true for rats treated with miR-146a agomir. Conclusion Our findings elucidate the role of inflammation in the pathogenesis of SE in immature rats, and show that regulating the expression level of miR-146a may provide a novel insights into the pathogenesis of SE.

Despite prompt and appropriate care, Streptococcus pneumoniae (S. pneumoniae) meningitis remains a key cause of childhood morbidity and mortality. Recently, several studies suggested the possibility of microRNAs (miRNA) involvement in multiple brain and infectious diseases. This study aimed to investigate the expressional changes of brain-enriched miRNAs (124, 134, 9, and 138), and inflammation-related miRNAs (132, 181a, 221, and 222) in the cerebral cortex of acute S. pneumoniae meningitis in postnatal day 25 rats with and without treatment. Quantitative polymerase chain reaction (qPCR) was used to measure the expression levels of the tested brain-enriched and inflammation-related miRNAs in the cerebral cortical tissues obtained from acute experimental meningitis rat model induced by intracranial inoculation of S. pneumoniae serotype 3 with and without treatment. Control rats inoculated with saline were also included. Brainenriched miRNAs are significantly downregulated in untreated animals while after treatment with antibiotic and steroid for 24 hours, miR-124 and miR-9 expressions were nearly equal to the control, while miR-134 and miR-138 were significantly upregulated. Inflammation-related miR-132 was significantly downregulated in untreated and treated animals while miRNAs (181a, 221, and 222) were significantly upregulated in untreated and treated animals. Acute S. pneumoniae meningitis leads to significant changes in the cerebral cortical expressions of some brain-enriched and inflammation-related miRNAs. These miRNAs may play a role in the pathogenesis of acute bacterial meningitis.

Herein we show the development of a minimally instrumented paper-based molecular diagnostic for point of care detection of sexually transmitted infections caused by Chlamydia trachomatis. This new diagnostic platform incorporates cell lysis, isothermal nucleic acid amplification, and lateral flow visual detection using only pressure and heat sources, eliminating the need for expensive laboratory equipment. This paper-based test can be performed in less than one hour and has a clinically relevant limit of detection that is 100× more sensitive than current rapid immunoassays used for chlamydia diagnosis.

Microglia plays important role in central nervous system immune surveillance and has emerged as an essential cellular component for understanding brain diseases. MicroRNAs (miRs) are small, noncoding RNAs that regulate the post-transcriptional expression of protein-coding mRNAs, which may have key roles in microglial activation in response to brain ischemia and other stressors. Primary cultured rat microglial cells were prepared, and then microglial activation model was established by oxygen-glucose deprivation (OGD) method. Morphological observation, CD11b/c immunofluorence, MTT assay and Propidium iodide staining were done to test microglia viability at different OGD time points (0, 5, 10, 15, 30, 60 min). qPCR were performed to detect the dynamic changes in expressions of inflammation-related miRs (146a, 21, 181a, 221, and 222) and brain-enriched miRs (124, 134, 9, 132, and 138) in resting microglia and after challenge with OGD for the same time points. The activation and viability of the microglia was time dependent. Similarly, expressions of different miRs in microglia were significantly upregulated and reached the peak at different time points before reaching the baseline level with extension of OGD. Our data demonstrates for the first time that OGD as a model of an ischemic insult modulates the expressions of some inflammation-related and brain-enriched miRs. These changes may help to explore the molecular basis of microglia activation on the post-transcriptional level in response to different time points of OGD.