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BACKGROUND Spinal manipulation therapy (SMT) has been widely used worldwide to treat musculoskeletal diseases, but it can cause serious adverse events. Spinal epidural hematoma (SEH) caused by SMT is a rare emergency that can cause neurological dysfunction. We herein report three cases of SEH after SMT. CASE SUMMARY The first case was a 30-year-old woman who experienced neck pain and numbness in both upper limbs immediately after SMT. Her symptoms persisted after 3 d of conservative treatment, and she was admitted to our hospital. Magnetic resonance imaging (MRI) demonstrated an SEH, extending from C6 to C7. The second case was a 55-year-old man with sudden back pain 1 d after SMT, numbness in both lower limbs, an inability to stand or walk, and difficulty urinating. MRI revealed an SEH, extending from T1 to T3. The third case was a 28-year-old man who suddenly developed symptoms of numbness in both lower limbs 4 h after SMT. He was unable to stand or walk and experienced mild back pain. MRI revealed an SEH, extending from T1 to T2. All three patients underwent surgery after failed conservative treatment. The three cases recovered to ASIA grade E on day 5, 1 wk, and day 10 after surgery, respectively. All patients returned to normal after 3 mo of follow-up. CONCLUSION SEH caused by SMT is very rare, and the condition of each patient should be evaluated in full detail before operation. SEH should be diagnosed immediately and actively treated by surgery.

Anthocyanins are water-soluble pigments in plants known for their photoprotective role against photoinhibitory and photooxidative damage under high light (HL). However, it remains unclear whether light-shielding or antioxidant activity plays a major role in the photoprotection exerted by anthocyanins under HL stress. To shed light on this question, we analyzed the physiological and biochemical responses to HL of three Arabidopsis thaliana lines (Col, chi, ans) with different light absorption and antioxidant characteristics. Under HL, ans had the highest antioxidant capacity, followed by Col, and finally chi; Col had the strongest light attenuation capacity, followed by chi, and finally ans. The line ans had weaker physiological activity of chloroplasts and more severe oxidative damage than chi after HL treatment. Col with highest photoprotection of light absorption capacity had highest resistance to HL among the three lines. The line ans with high antioxidant capacity could not compensate for its disadvantages in HL caused by the absence of the light-shielding function of anthocyanins. In addition, the expression level of the Anthocyanin Synthase (ANS) gene was most upregulated after HL treatment, suggesting that the conversion of colorless into colored anthocyanin precursors was necessary under HL. The contribution of anthocyanins to flavonoids, phenols, and antioxidant capacity increased in the late period of HL, suggesting that plants prefer to synthesize red anthocyanins (a group of colored antioxidants) over other colorless antioxidants to cope with HL. These experimental observations indicate that the light attenuation role of anthocyanins is more important than their antioxidant role in photoprotection.

Objective Paired box 7 ( PAX7) has been considered as a candidate gene for non-syndromic cleft lip with or without palate (NSCL/P). However, there is no research for the XXX, and previous studies concentrated on limited variants. This study aimed to conduct sufficiently dense and powerful scans of variants at PAX7 and explored the roles of variants at PAX7 in NSCL/P among the XXX. Design Targeted region sequencing was performed to thoroughly screen variations, followed by a two-phase association analysis. 159 NSCL/P cases and 542 controls were analyzed in phase 1. Then in phase 2, the validation study was performed using 1626 cases and 2255 controls. We also explored the roles of variants at PAX7 gene in NSCL/P subtypes. Additionally, indirect associations were found by calculating LD and haplotypes. Setting The study was conducted in XXX. Patients, participants 159 NSCL/P cases and 542 controls were analyzed in phase 1. Then in phase 2, the validation study was performed using 1626 cases and 2255 controls. Interventions Blood samples were collected. Main outcome measures To explore the association analysis between variants at PAX7 and NSCL/P in XXX. Results The results showed that rs2236810, rs114882979 and rs2236804 were significantly associated with NSCL/P, which were predicted to have regulatory functions. Besides, variants at PAX7 function differently in the NSCL/P subtypes. We also discovered a PAX7 missense variant, NM_001135254 p.A369 V (NM_002584.2:c.1106C > T). Conclusions In summary, we confirmed 3 SNPs at PAX7 were significantly associated with NSCL/P in XXX and identified a missense variant, NM_001135254 p.A369 V (NM_002584.2:c.1106C > T).

The purpose of this study is to analyze the clinical characteristics of patients with Van der Woude syndrome (VWS) and to detect variations in each patient. Finally, the combination of genotype and phenotype can make a clear diagnosis of VWS patients with different phenotype penetrance. Five Chinese VWS pedigree were enrolled. Whole exome sequencing of the proband was performed, and the potential pathogenic variation was further verified by Sanger sequencing in the patient and their parents. The human mutant IRF6 coding sequence was generated from the human full-length IRF6 plasmid by site-directed mutagenesis and cloned into the GV658 vector, RT-qPCR and Western blot were used to detect the expression of IRF6. We found one de novo nonsense variation (p. Gln118Ter) and three novel missense variations (p. Gly301Glu, p. Gly267Ala, and p. Glu404Gly) co-segregated with VWS. RT-qPCR analysis revealed that p. Glu404Gly significantly reduced the expression level of IRF6 mRNA. Western blot of cell lysates confirmed that IRF6 p. Glu404Gly abundance levels were lower than those for IRF6 wild type. This discovery of the novel variation ( IRF6 p. Glu404Gly) expands the spectrum of known variations in VWS in Chinese humans. Genetic results combined with clinical phenotypes and differential diagnosis points from other diseases can make a definitive diagnosis and provide genetic counseling for families.

Purpose Carpal tunnel syndrome is one of the common peripheral neuropathies. For magnetic resonance imaging, segmentation of the carpal tunnel and its contents, including flexor tendons and the median nerve for magnetic resonance images is an important issue. In this study, a convolutional neural network (CNN) model, which was modified by the original DeepLabv3 + model to segment three primary structures of the carpal tunnel: the carpal tunnel, flexor tendon, and median nerve. Methods To extract important feature maps for segmentation of the carpal tunnel, flexor tendon, and median nerve, the proposed CNN model termed modified DeepLabv3 + uses DenseNet-121 as a backbone and adds dilated convolution to the original spatial pyramid pooling module. A MaskTrack method was used to refine the segmented results generated by modified DeepLabv3 + , which have a small and blurred appearance. For evaluation of the segmentation results, the average Dice similarity coefficients (ADSC) were used as the performance index. Results Sixteen MR images corresponding to different subjects were obtained from the National Cheng Kung University Hospital. Our proposed modified DeepLabv3 + generated the following ADSCs: 0.928 for carpal tunnel, 0.872 for flexor tendons and 0.785 for the median nerve. The ADSC value of 0.8053 generated the MaskTrack that 0.22 ADSC measure were improved for measuring the median nerve. Conclusions The experimental results showed that the proposed modified DeepLabv3 + model can promote segmentations of the carpal tunnel and its contents. The results are superior to the results generated by original DeepLabv3 + . Additionally, MaskTrack can also effectively refine median nerve segmentations.

Ligament tissue engineering is currently a novel approach to the treatment of ligament injury, which can replace the deficiency of autografts. Ligament tissue engineering consists of four basic elements:seed cells, nanoscaffolds, growth factors, and mechanical stimulation. At present, the main problem in ligament tissue engineering is how to control seed cells to ligament cells more controllly. The study found that each physical property of the natural bio ligament and mechanical stimulation (uniaxial stretching) plays an important role in the differentiation of stem cells into ligament cells. Therefore, the design of nanofiber scaffolds must consider the elastic modulus of the material and the material. Structure(material arrangement, porosity and diameter, etc.), elastic modulus and material structure in different ranges will guide cells to differentiate into different lineages. Considering that the ligament is the main force-bearing tissue of the human body, mechanical stimulation is also essential for stem cell differentiation, especially uniaxial stretching, which best meets the stress of the ligament in the body. A large number of studies have found the frequency and amplitude of stretching. And time will also lead the cells to differentiate in different directions. RhoA/ROCK plays a regulatory role in cytoskeletal remodeling and cell differentiation. It is also found that RhoA/ROCK protein participates in the process of nanofiber arrangement and uniaxial stretching to guide stem cells to differentiate into ligament cells, specifically how to influence stem cell differentiation. It is not clear at present that understanding the effects of physical properties on stem cell differentiation and understanding the mechanism of action of RhoA/ROCK protein will provide a new theoretical basis for further optimization of ligament tissue engineering.

Tylophorine-based compounds and natural cardiotonic steroids (cardenolides and bufadienolides) are two classes of transmissible gastroenteritis coronavirus inhibitors, targeting viral RNA and host cell factors, respectively We tested both types of compounds against two types of coronaviruses, to compare and contrast their antiviral properties, and with view to their further therapeutic development Examples of both types of compounds potently inhibited the replication of both feline infectious peritonitis virus and human coronavirus OC43 with EC50 values of up to 8 and 16 nM, respectively Strikingly, the tylophorine-based compounds tested inhibited viral yields of HCoV-OC43 to a much greater extent (7–8 log magnitudes of p f u /ml) than the cardiotonic steroids (about 2–3 log magnitudes of p f u /ml), as determined by end point assays Based on these results, three tylophorine-based compounds were further examined for their anti-viral activities on two other human coronaviruses, HCoV-229E and SARS-CoV-2 These three tylophorine-based compounds inhibited HCoV-229E with EC50 values of up to 6 5 nM, inhibited viral yields of HCoV-229E by 6–7 log magnitudes of p f u /ml, and were also found to inhibit SARS-CoV-2 with EC50 values of up to 2 5–14 nM In conclusion, tylophorine-based compounds are potent, broad-spectrum inhibitors of coronaviruses including SARS-CoV-2, and could be used for the treatment of COVID-19 © Copyright © 2020 Yang, Lee, Hsu, Jan, Lin, Chang, Peng, Yang, Liang, Liao, Chao, Pang, Kao, Huang, Lin, Chang, Niu, Wu, Sytwu, Chen and Lee

This paper mainly develops a robust adaptive tracking control scheme to counter effects caused by load fluctuation and external disturbances. An adaptive controller based on appropriate control strategy and adaptive law is presented based on Lyapunov stability theory. The output voltage asymptotic tracking of the buck converters is obtained even under the influence of load fluctuation and external disturbances. Simulation results also reveal the availability of the proposed adaptive control scheme.

Danshen, the dried rhizome of Salvia miltiorrhiza Bunge is a widely used herb in traditional Chinese medicinal preparations to treat cardiovascular diseases, renal and liver diseases. Tanshinones and salvianolic acids are used as markers to survey the commercial available products. In this study, a convenient high-performance liquid chromatographic method was developed to simultaneously separate and identify six main polyphenolic components, caffeic acid (1), danshensu (2), lithospermic acid (3), rosmarinic acid (4), salvianolic acid B (5) and salvianolic acid A (6), and four major abietane-type diterpenes, 15,16-dihydrotanshinone I (7), cryptotanshinone (8), tanshinone I (9), and tanshinone IIA (10), with UV and MS detectors by comparing their retention time, MS and MS2 data with those data obtained from the authentic compounds. A good resolution of all the analytes was obtained by choosing methanol and 0.25% aqueous acetic acid (adjusted to pH 2.77 with 1N HCl) as the mobile phase. By using LC-UV (290 nm)-MS method, polyphenols (1-6) and tanshinones (7-10) were easily detected and unambiguously identified in commercial Danshen.

Objectives This study aimed to initially screen more susceptible variations at PAX9 gene in non-syndromic cleft palate only cases and then test their associations in independent non-syndromic orofacial cleft subtypes from Western Han Chinese population. Design Initially exons at PAX9 gene were screened for variations among 180 non-syndromic cleft palate only patients by the Sanger sequencing method, and the genotype data from 1000 Genomes Project Database was taken as control to perform Chi-square test. Subsequently, we performed association analysis among 2202 non-syndromic orofacial cleft patients (938 non-syndromic cleft palate only cases, 456 non-syndromic cleft lip only cases and 808 non-syndromic cleft lip and palate cases) and 1823 normal controls to replicate the role of two significant single nucleotide polymorphisms. Results Two intronic single nucleotide polymorphism rs12885612 (P = 0.047, OR = 0.671 and 95−0.996) and rs12881248 (P = 0.047, OR = 1.006 and 95 %CI: 0.443−0.995) were associated non-syndromic cleft palate only in the initial screen phase. Surprisingly, in replication analysis rs12885612 (P = 0.048, OR = 1.25, 95 %CI: 1.0–1.56) and rs12881248 (P = 0.037, OR = 1.26, 95 %CI: 1.01−1.57) exhibited significant association signal in non-syndromic cleft lip only group, but not in non-syndromic cleft palate only group. Conclusion In summary, this study firstly revealed that rs12885612 and rs12881248 at PAX9 gene associated with non-syndromic cleft lip only from Western Han Chinese population, which indicated that PAX9 is a promising susceptible gene for non-syndromic cleft lip only in Western Han Chinese population.

A series of naturally occurring cardenolides that exhibit potent anti-transmissible gastroenteritis virus (TGEV) activity in swine testicular (ST) cells has been identified. In an immunofluorescence assay, these cardenolides were found to diminish the expressions of TGEV nucleocapsid and spike protein, which was used as an indication for viral replication; block TGEV infection induced apoptosis and cytopathic effects; and impart the same trend of inhibitory activity against Na+/K+-ATPase as for anti-TGEV activity. The viral titer inhibition was found to take place in a dose-dependent manner. Knocking down expression of Na+/K+-ATPase, the cellular receptor of cardenolides, in ST cells was found to significantly impair the susceptibility of ST cells to TGEV infectivity. Thus, we have identified Na+/K+-ATPase as an anti-viral drug target and its antagonists, cardenolides, a novel class of anti- TGEV agents.

Humans have two glutaminase genes, GLS (GLS1) and GLS2, each of which has two alternative transcripts: the kidney isoform (KGA) and glutaminase C (GAC) for GLS, and the liver isoform (LGA) and glutaminase B (GAB) for GLS2. Initial hit compound (Z)-5-((1-(4-bromophenyl)-2,5-dimethyl-1H-pyrrol-3-yl)methylene)thiazolidine-2,4-dione (2), a thiazolidine-2,4-dione, was obtained from a high throughput screening of 40 000 compounds against KGA. Subsequently, a series of thiazolidine-2,4-dione derivatives was synthesized. Most of these were found to inhibit KGA and GAC with comparable activities, were less potent inhibitors of GAB, and were moderately selective for GLS1 over GLS2. The relationships between chemical structure, activity, and selectivity were investigated. The lead compounds obtained were found to (1) offer in vitro cellular activities for inhibiting cell growth, clonogenicity, and cellular glutamate production, (2) exhibit high concentrations of exposure in plasma by a pharmacokinetic study, and (3) reduce the tumor size of xenografted human pancreatic AsPC-1 carcinoma cells in mice.

On-board diagnostics II (OBD-II) [1]is a new automotive device, which can provide the vehicle's self-diagnostic and reporting capability to the vehicle owner or repair technician. And the vehicle owner or repair technician can understand the status of the various vehicle subsystems. These vehicle's self-diagnostic reports are very basic material for the car big data. Currently marketed OBD-II system can only be used for small cars. However, the large vehicle is the main cause of car accident. Therefore, the proposed OBD-II device is suitable for the niche market of the bus and commercial vehicles.

Interruption of the Warburg effect - the observation that un-stimulated macrophages reprogram their core metabolism from oxidative phosphorylation toward aerobic glycolysis to become pro-inflammatory M1 macrophages upon stimulation - is an emerging strategy for the treatment of cancer and anti-inflammatory diseases such as rheumatoid arthritis. We studied this process with view to the discovery of novel therapeutics, and found that tylophorine-based compounds targeted a ribonucleoprotein complex containing caprin-1 and mRNAs of c-Myc and HIF-1α in LPS/IFN-γ stimulated Raw264.7 cells, diminished the protein levels of c-Myc and HIF-1α, and consequently downregulated their targeted genes that are associated with the Warburg effect, as well as the pro-inflammatory iNOS and COX2. The tylophorine-based compound DBQ 33b significantly meliorated the severity and incidence of type II collagen-monoclonal antibody-induced rheumatoid arthritis and diminished gene expressions of c-Myc, HIF-1α, iNOS, COX2, TNFα, and IL-17A in vivo. Moreover, pharmacological inhibition of either c-Myc or HIF-1α exhibited similar effects as the tylophorine-based compound DBQ 33b, even though inhibition of c-Myc reversed the induction of iNOS and COX2 in LPS/IFN-γ stimulated Raw264.7 cells to a lesser degree. Therefore, simultaneous inhibition of both c-Myc and HIF-1α is efficacious for anti-inflammation in vitro and in vivo and merits further study.

Carpal tunnel syndrome commonly occurs in individuals working in occupations that involve use of vibrating manual tools or tasks with highly repetitive and forceful manual exertion. In recent years, carpal tunnel syndrome has been evaluated by ultrasound imaging that monitors median nerve movement. Conventional image analysis methods, such as the active contour model, are typically used to expedite automatic segmentation of the median nerve, but these usually suffer from an arduous manual intervention. We propose a new convolutional neural network framework for localization and segmentation of the median nerve, called DeepNerve, that is based on the U-Net model. DeepNerve integrates the characteristics of MaskTrack and convolutional long short-term memory to effectively locate and segment the median nerve. On the basis of experimental results, the proposed model achieved high performance and generated average Dice measurement, precision, recall and F-score values of 0.8975, 0.8912, 0.9119 and 0.9015, respectively. The segmentation results of DeepNerve were significantly improved in comparison with those of conventional active contour models. Additionally, the results of Student's t-test revealed significant differences in four deformation measurements of the median nerve, including area, perimeter, aspect ratio and circularity. We conclude that the proposed DeepNerve not only generates satisfactory results for localization and segmentation of the median nerve, but also creates more promising measurements for applications in clinical carpal tunnel syndrome diagnosis.

This paper presents a compensation circuitry design method for a class of linear time-invariant systems with bounded time-varying disturbances. An adaptive compensation control strategy is constructed to eliminate the effects of disturbances based on signals from an adaption mechanism. Through Lyapunov stability theory, asymptotic/exponential stability results are obtained for the closed-loop systems under the influence of time-varying disturbances. Furthermore, the proposed adaptive control strategy is physically implemented by circuits based on circuit theory. A numerical example and its Multisim simulation results are given to support the theoretical findings.

Colorimetric gene detection based on gold nanoparticles (AuNPs) is an attractive detection format due to its simplicity. Here, we report a new design for a colorimetric gene-sensing platform based on the CRISPR/Cas system that has improved specificity, sensitivity, and universality. CRISPR/Cas12a and CRISPR/Cas13a have two distinct catalytic activities and are used for specific target gene recognition. Programmable recognition of DNA by Cas12a/crRNA and RNA by Cas13a/crRNA with a complementary sequence activates the nonspecific trans-ssDNA or -RNA cleavage, respectively, thus degrading the ssDNA or RNA linkers which are designed as a hybridization template for the AuNP-DNA probe pair. Target-induced trans -ssDNA or RNA cleavage leads to a distance-dependent color change for the AuNP-DNA probe pair. In this platform, naked eye detection of transgenic rice, African swine fever virus (ASFV), and a miRNA can be completed within 1 hour. Our colorimetric gene-sensing method shows superior characteristics, such as probe universality, isothermal reaction conditions, on-site detection capability, and sensitivity that is comparable to that of the fluorescent detection; thus, this method represents a robust next generation gene detection platform.

An unprecedented biological function of natural cardenolides independent of their membrane target Na

Graphical abstract, Highlights • Natural cardenolides suppress coronaviral activity via downregulating JAK1. • Natural cardenolides downregulate JAK1 in a Na+/K+-ATPase independent manner. • Ouabain (cardenolides) activates Ndfip1/2 and NEDD4 to mediate JAK1 proteolysis. • Ndfip1/2 and NEDD4 mediated JAK1 proteolysis attenuates coronaviral activity., An unprecedented biological function of natural cardenolides independent of their membrane target Na+/K+-ATPase is disclosed. Previously, we reported that cardenolides impart anti-transmissible gastroenteritis coronavirus (anti-TGEV) activity through the targeting of Na+/K+-ATPase and its associated PI3K_PDK1_RSK2 signaling. Swine testis cells with Na+/K+-ATPase α1 knocked down exhibited decreased susceptibility to TGEV infectivity and attenuated PI3K_PDK1_RSK2 signaling. Herein, we further explored a Na+/K+-ATPase-independent signaling axis induced by natural cardenolides that also afforded significant anti-coronaviral activity for porcine TGEV and human HCoV-OC43. Using pharmacological inhibition and gene silencing techniques, we found that this anti-TGEV or anti-HCoV-OC43 activity was caused by JAK1 proteolysis and mediated through upstream activation of Ndfip1/2 and its effector NEDD4. This study provides novel insights into the pharmacological effects of natural cardenolides, and is expected to inform their future development as antiviral agents.

Introductory video games have become an attractive form of entertainment and have surpassed television viewing as a preferred sedentary activity choice for users. Introductory video games are more and more popular for users studying E-learning topics. Atomic energy is important in the fields of science education, industry and research. However, the lack of a thorough understanding of nuclear energy and radiation, and the attitudes towards the safety of nuclear power are important issues. This paper discusses the teaching of the fundamental knowledge of nuclear power to students, to enhance their understanding of including nuclear power and safety concerns. This objective was achieved by means of e-learning which contains a variety of information about general science. This plan established a Nuclear Energy 3D eBook by integrating an eBook and TV devices, using the Tsing Hua Open-pool Reactor (THOR) as an example. We employed a nuclear energy education design in a serious game. We recruited N users to participate in THOR nuclear energy education learning. The present study examines the usage pattern of electronic game devices among university students in Taiwan. A survey was conducted among 31 students in a university, with 17 males and 14 females in the age range of 19–21. The results showed students can clarify their ideas about nuclear power by using the materials. The impact of nuclear energy 3D eBook on users who learn using the 3D teaching materials was also examined.

Na+/K+-ATPase α1 was reported to directly interact with and recruit FGF2 (fibroblast growth factor 2), a vital cell signaling protein implicated in angiogenesis, to the inner plasma membrane for subsequent secretion. Cardenolides, a class of cardiac glycosides, were reported to downregulate FGF2 secretion upon binding to Na+/K+-ATPase α1 in a cell system with ectopically expressed FGF2 and Na+/K+-ATPase α1. Herein, we disclose that the cardenolides ouabain and reevesioside A significantly enhance the secretion/release of FGF2 and the phosphorylation of FGFR1 (fibroblast growth factor receptor 1) in a time- and dose-dependent manner, in A549 carcinoma cells. A pharmacological approach was used to elucidate the pertinent upstream effectors. Only the ERK1/2 inhibitor U0126 but not the other inhibitors examined (including those inhibiting the unconventional secretion of FGF2) was able to reduce ouabain-induced FGF2 secretion and FGFR1 activation. ERK1/2 phosphorylation was increased upon ouabain treatment, a process found to be mediated through upstream effectors including ouabain-induced phosphorylated EGFR and a reduced MKP1 protein level. Therefore, at least two independent lines of upstream effectors are able to mediate ouabain-induced ERK1/2 phosphorylation and the subsequent FGF2 secretion and FGFR1 activation. These finding constitute unprecedent insights into the regulation of FGF2 secretion by cardenolides.

Leaf senescence is an important factor that affects crop yield traits and is regulated by various factors. Here, we propose the photo–carbon imbalance hypothesis to explain the mechanism of rice leaf senescence. The main idea of this hypothesis is that carbon assimilation decreases faster than the absorption of light energy in photosynthesis during the late stages of rice growth, which ultimately results in leaf senescence. Our results indicate that endogenous ascorbic acid (Asc) plays an important role in leaf senescence by affecting the expression of senescence genes, thereby influencing photosynthetic capacity and consequently grain yield. The effects of exogenous Asc and methyl jasmonate (MeJA) on photosynthetic capability implied that the balance between photoreaction and carbon assimilation is regulated by exogenous antioxidants or accelerators of senescence. The results of the shading treatments indicated that shading will mitigate the photo–carbon imbalance and improve photosynthetic capacity, resulting in increased yields. Increasing antioxidant concentrations can enhance the reactive oxygen species (ROS) scavenging capacity, whereas shading reduces excess light energy, which may help to restore the photo–carbon balance.

// Ya-Qi Qiu 1,2 , Cheng-Wei Yang 1 , Yue-Zhi Lee 1 , Ruey-Bing Yang 3 , Chih-Hao Lee 4 , Hsing-Yu Hsu 1 , Chien-Chung Chang 2 and Shiow-Ju Lee 1 1 Institute of Biotechnology and Pharmaceutical Research, National Health Research Institutes, Miaoli, Taiwan 2 Graduate Program of Biotechnology in Medicine, Institute of Molecular & Cellular Biology, National Tsing Hua University, Hsinchu, Taiwan 3 Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan 4 Department of Genetics and Complex Diseases, Division of Biological Sciences, Harvard School of Public Health, Boston, Massachusetts, USA Correspondence: Shiow-Ju Lee, email: // Keywords : Caprin-1, c-Myc, Cyclin D, G3BP1, Processing Body Received : October 02, 2014 Accepted : December 09, 2014 Published : December 10, 2014 Abstract Tylophorine compounds have been the focus of drug development for decades. Tylophorine derivatives exhibit anti-cancer activities but their cellular targets remain unknown. We used a biotinylated tylophorine derivative to probe for the interacting cellular target(s) of tylophorine. Tylophorine directly binds to caprin-1 and consequently enhances the recruitment of G3BP1, c-Myc mRNA, and cyclin D2 mRNA to form a ribonucleoprotein complex. Subsequently, this tylophorine targeted ribonucleoprotein complex is sequestered to the polysomal fractions and the protein expressions of the associated mRNA-transcripts are repressed. Caprin-1 depleted carcinoma cells become more resistant to tylophorine, associated with decreased formation of the ribonucleoprotein complex targeted by tylophorine. Consequently, tylophorine downregulates c-Myc and cyclins D1/D2, causing hypophosphorylation of Rb and suppression of both processing-body formation and the Warburg effect. Gene expression profiling and gain-of-c-Myc-function experiments also revealed that the downregulated c-Myc contributes to the anti-oncogenic effects of tylophorine compounds. Furthermore, the potent tylophorine derivative dibenzoquinoline-33b elicited a similar effect, as c-Myc protein levels were also decreased in xenograft tumors treated with dibenzoquinoline-33b. Thus, tylophorine compounds exert anti-cancer activity predominantly by targeting and sequestering the caprin-1 protein and c-Myc mRNA associated ribonucleoprotein complex.

// Yue-Zhi Lee 1,* , Cheng-Wei Yang 1,* , Hsin-Yu Chang 1,* , Hsing-Yu Hsu 1,* , Ih-Shen Chen 2 , Hsun-Shuo Chang 2 , Chih-Hao Lee 3 , Jinq-chyi Lee 1 , Chidambaram Ramesh Kumar 1 , Ya-Qi Qiu 1,4 , Yu-Sheng Chao 1 and Shiow-Ju Lee 1 1 Institute of Biotechnology and Pharmaceutical Research, National Health Research Institutes, Miaoli, Taiwan 2 School of Pharmacy, College of Pharmacy, Kaohsiung Medical University, Kaohsiung, Taiwan 3 Department of Genetics and Complex Diseases, Division of Biological Sciences, Harvard School of Public Health, Boston, Massachusetts, USA 4 Graduate Program of Biotechnology in Medicine, Institute of Molecular & Cellular Biology, National Tsing Hua University, Hsinchu, Taiwan * These authors equally contributed to this work Correspondence: Shiow-Ju Lee, email: // Keywords : AMPK; Autophagy; Glutaminase; mTOR; Raptor; ULK1 Received : April 26, 2014 Accepted : July 7, 2014 Published : July 8, 2014 Abstract Glutaminase, which converts glutamine to glutamate, is involved in Warburg effect in cancer cells. Two human glutaminase genes have been identified, GLS ( GLS1 ) and GLS2 . Two alternative transcripts arise from each glutaminase gene: first, the kidney isoform (KGA) and glutaminase C (GAC) for GLS ; and, second, the liver isoform (LGA) and glutaminase B (GAB) for GLS2 . While GLS1 is considered as a cancer therapeutic target, the potential role of GLS2 in cancer remains unclear. Here, we discovered a series of alkyl benzoquinones that preferentially inhibit glutaminase B isoform (GAB, GLS2) rather than the kidney isoform of glutaminase (KGA, GLS1). We identified amino acid residues in an allosteric binding pocket responsible for the selectivity. Treatment with the alkyl benzoquinones decreased intracellular glutaminase activity and glutamate levels. GLS2 inhibition by either alkyl benzoquinones or GLS2 siRNA reduced carcinoma cell proliferation and anchorage-independent colony formation, and induced autophagy via AMPK mediated mTORC1 inhibition. Our findings demonstrate amino acid sequences for selective inhibition of glutaminase isozymes and validate GLS2 as a potential anti-cancer target.

This paper presents the interval dynamic analysis of structures with uncertain-but-bounded parameters under stochastic process excitations. Structural natural frequencies and mean square values of structural random responses are not deterministic values but intervals. The interval problems are converted to optimization problems. Mathematical models are developed to find the bounds of interval natural frequencies and mean square displacements. An improved particle swarm optimization algorithm, namely lower sequence initialized high-order nonlinear particle swarm optimization algorithm, is employed to capture the exact bounds of structural dynamic characteristics and random vibration responses. Numerical example is used to demonstrate the presented method. Quasi-Monte Carlo simulations are also implemented to validate the change ranges of structural natural frequencies and mean square displacements produced by the proposed method.