In vitro therapeutic effects and molecular mechanisms of targeted inhibition of CDK12/13 in high-grade gliomas

Objective·To find novel and common targeting strategies for high-grade gliomas (HGGs) from the perspective of epigenetic and transcriptional modulators, test the therapeutic effect in vitro and investigate the related molecular mechanisms.Methods·Glioblastoma (GBM) and diffuse intrinsic pontine glioma (DIPG) cell lines with high malignancy and mortality in HGGs were selected for screening of targeted small molecule drug library related to epigenetic transcription and for functional genome screening based on the CRISPR-Cas9 technology. The effect of selected targeted epigenetic transcriptional modulators on growth, proliferation, and apoptosis of GBM and DIPG cell lines were then measured either by CRISPR-Cas9 knockout or treatment with targeted small molecule inhibitors of genes in vitro. Anti-tumor molecular mechanisms of the modulators in corresponding small molecule inhibitors-treated GBM and DIPG cells were explored via RNA-seq transcriptome analysis and further verified by real time quantitative PCR (RT-qPCR), Western blotting and flow cytometry.Results·Targeted small molecule drug library combined with functional genome screening for epigenetic transcriptional modulators identified CDK12/13 as the novel therapeutic targets for both GBM and DIPG. Knockout out of CDK12 by CRISPR-Cas9 in multiple GBM and DIPG cell lines significantly reduced their in vitro cellular activity. CDK12/13 inhibitors SR-4835 and THZ531 also significantly inhibited the growth of these two types of HGGs cell lines in vitro by antagonizing cell proliferation and promoting cell apoptosis. RNA-seq transcriptome analysis of GBM and DIPG cell lines after SR-4835 treatment showed that genes significantly down-regulated by CDK12/13 inhibitors in HGGs cells were mainly enriched in transcriptional regulation, DNA damage response (DDR) pathway, ubiquitin-proteasome pathway, and cell cycle. Furthermore, a series of experiments demonstrated that targeted inhibition of CDK12/13 significantly down-regulated the transcription of DDR-related genes, resulting in the accumulation of DNA damage, and induced G2-M cell cycle arrest.Conclusion·CDK12/13 is a common potential therapeutic target of these two types of HGGs, providing theoretical support for the follow-up in vivo verification and combination therapy test. The research also lays the foundation for further clinical application

Screening and construction of nanobodies against human CD93 using phage libraries and study of their antiangiogenic effects

BackgroundCluster of Differentiation 93 (CD93) plays an important role in angiogenesis and is considered an important target for inhibiting tumor angiogenesis, but there are currently no therapeutic antibodies against CD93 in the clinic. Thus, we describe the screening of novel nanobodies (Nbs) targeting human CD93 from a phage library of shark-derived Nbs.MethodsScreening and enrichment of phage libraries by enzyme-linked immunosorbent assay (ELISA). Anti-CD93 Nbs were purified by expression in E. coli. The binding affinity of anti-CD93 Nbs NC81/NC89 for CD93 was examined by flow cytometry (FC) and ELISA. The thermal stability of NC81/NC89 was examined by ELISA and CD spectroscopy. Afterward, the anti-angiogenic ability of NC81/NC89 was examined by MTT, wound healing assay, and tube formation assay. The expression level of VE-cadherin (VE-Ca) and CD93 was detected by Western Blot (WB). The binding sites and binding forms of NC81/NC89 to CD93 were analyzed by molecular docking.ResultsThe anti-CD93 Nbs were screened in a phage library, expressed in E. coli, and purified to >95% purity. The results of FC and ELISA showed that NC81/NC89 have binding ability to human umbilical vein endothelial cells (HUVECs). The results of ELISA and CD spectroscopy showed that NC81/NC89 retained the ability to bind CD93 at 80°C and that the secondary structure remained stable. In vitro, the results showed that NC81 and NC89 significantly inhibited the proliferation and migration of human umbilical vein endothelial cells (HUVECs) as well as tube formation on Matrigel. Western Blot showed that NC81 and NC89 also inhibited the expression of VE-Ca thereby increasing vascular permeability. It was found during molecular docking that the CDR regions of NC81 and NC89 could be attached to CD93 by strong hydrogen bonds and salt bridges, and the binding sites were different.ConclusionWe have successfully isolated NC81 and NC89, which bind CD93, and both Nbs significantly inhibit angiogenesis and increase vascular permeability. These results suggest that NC81 and NC89 have potential clinical applications in angiogenesis-related therapies

Progress of the special-subjects study on the construction of comprehensive geological disaster prevention and control system in Yunnan Province

The establishment of the comprehensive geological disaster prevention and control system in Yunnan province stands as China’s most extensive and grand-scale endeavor in safeguarding the prevention and control of geological disasters in China. Its implementation has led to a significant reduction in the occurrence of geological disasters, resulting in a substantial decrease in both casualties and missing persons affected by such disasters. This accomplishment has yielded remarkable outcomes in disaster prevention and mitigation. Based on the results of 12 series monographic studies, this paper provides an overview of the implementation of the system and its disaster prevention and mitigation effects. It summarizes the main scientific and technological achievements, with a particular focus on the causes and patterns of plateau geological disasters, understanding of special rock and soil disaster control mechanisms, susceptibility zoning evaluation, comprehensive remote sensing identification of geological hazards, progress in automated monitoring and early warning, and the development of geological environment information standard system. These achievements can provide valuable insights for the comprehensive geological disaster prevention and control in Yunnan Province

Novel online data allocation for hybrid memories on tele-health systems

[EN] The developments of wearable devices such as Body Sensor Networks (BSNs) have greatly improved the capability of tele-health industry. Large amount of data will be collected from every local BSN in real-time. These data is processed by embedded systems including smart phones and tablets. After that, the data will be transferred to distributed storage systems for further processing. Traditional on-chip SRAMs cause critical power leakage issues and occupy relatively large chip areas. Therefore, hybrid memories, which combine volatile memories with non-volatile memories, are widely adopted in reducing the latency and energy cost on multi-core systems. However, most of the current works are about static data allocation for hybrid memories. Those mechanisms cannot achieve better data placement in real-time. Hence, we propose online data allocation for hybrid memories on embedded tele-health systems. In this paper, we present dynamic programming and heuristic approaches. Considering the difference between profiled data access and actual data access, the proposed algorithms use a feedback mechanism to improve the accuracy of data allocation during runtime. Experimental results demonstrate that, compared to greedy approaches, the proposed algorithms achieve 20%-40% performance improvement based on different benchmarks. (C) 2016 Elsevier B.V. All rights reserved.This work is supported by NSF CNS-1457506 and NSF CNS-1359557.Chen, L.; Qiu, M.; Dai, W.; Hassan Mohamed, H. (2017). Novel online data allocation for hybrid memories on tele-health systems. Microprocessors and Microsystems. 52:391-400. https://doi.org/10.1016/j.micpro.2016.08.003S3914005

Jia-Wei-Kai-Xin-San treatment alleviated mild cognitive impairment through anti-inflammatory and antiapoptotic mechanisms in SAMP8 mice

Background. Alleviating mild cognitive impairment (MCI) is crucial to delay the progression of Alzheimer’s disease (AD). Jia-Wei-Kai-Xin-San (JWKXS) is applied for treating AD with MCI. However, the mechanism of JWKXS in the treatment of MCI is unclear. Thus, this study aimed to investigate the effect and mechanism of JWKXS in SAMP8 mice models of MCI. Methods. MCI models were established to examine learning and memory ability and explore the pathomechanisms in brain of SAMP8 mice at 4, 6, and 8 months. The mice were treated for 8 weeks and the effects of JWKXS on MCI were characterized through Morris water maze and HE/Nissl’s/immunohistochemical staining. Its mechanism was predicted by the combination of UPLC-Q-TOF/MS and system pharmacology analysis, further verified with SAMP8 mice, BV2 microglial cells, and PC12 cells. Results. It was found that 4-month-old SAMP8 mice exhibited MCI. Two months of JWKXS treatment improved the learning and memory ability, alleviated the hippocampal tissue and neuron damage. Through network pharmacology, four key signaling pathways were found to be involved in treatment of MCI by JWKXS, including TLR4/NF-κB pathway, NLRP3 inflammasome activation, and intrinsic and extrinsic apoptosis. In vitro and in vivo experiments demonstrated that JWKXS attenuated neuroinflammation by inhibiting microglia activation, suppressing TLR4/NF-κB and NLRP3 inflammasome pathways, and blocking the extrinsic and intrinsic apoptotic pathways leading to neuronal apoptosis suppression in the hippocampus. Conclusion. JWKXS treatment improved the learning and memory ability and conferred neuroprotective effects against MCI by inducing anti-inflammation and antiapoptosis. Limitations. The small sample size and short duration of the intervention limit in-depth investigation of the mechanisms. Future Prospects. This provides a direction for further clarification of the anti-AD mechanism, and provides certain data support for the formulation to move toward clinical practice

Improving feature location using structural similarity and iterative graph mapping

Locating program element(s) relevant to a particular feature is an important step in efficient maintenance of a software system. The existing feature location techniques analyze each feature independently and perform a one-time analysis after being provided an initial input. As a result, these techniques are sensitive to the quality of the input. In this paper, we propose to address the above issues in feature location using an iterative context-aware approach. The underlying intuition is that features are not independent of each other, and the structure of source code resembles the structure of features. The distinguishing characteristics of the proposed approach are: (1) it takes into account the structural similarity between a feature and a program element to determine feature-element relevance; (2) it employs an iterative process to propagate the relevance of the established mappings between a feature and a program element to the neighboring features and program elements. We evaluate our approach using two different systems, DirectBank, a small-scale industry financial system, and Linux kernel, a large-scale open-source operating system. Our evaluation suggests that the proposed approach is more robust and can significantly increase the recall of feature location with only a minor decrease of precision