Machine vision-based autonomous road hazard avoidance system for self-driving vehicles.

The resolution of traffic congestion and personal safety issues holds paramount importance for human's life. The ability of an autonomous driving system to navigate complex road conditions is crucial. Deep learning has greatly facilitated machine vision perception in autonomous driving. Aiming at the problem of small target detection in traditional YOLOv5s, this paper proposes an optimized target detection algorithm. The C3 module on the algorithm's backbone is upgraded to the CBAMC3 module, introducing a novel GELU activation function and EfficiCIoU loss function, which accelerate convergence on position loss lbox, confidence loss lobj, and classification loss lcls, enhance image learning capabilities and address the issue of inaccurate detection of small targets by improving the algorithm. Testing with a vehicle-mounted camera on a predefined route effectively identifies road vehicles and analyzes depth position information. The avoidance model, combined with Pure Pursuit and MPC control algorithms, exhibits more stable variations in vehicle speed, front-wheel steering angle, lateral acceleration, etc., compared to the non-optimized version. The robustness of the driving system's visual avoidance functionality is enhanced, further ameliorating congestion issues and ensuring personal safety.

Comparative Analysis of Clinical Outcomes: Posterior Cervical Endoscopic Discectomy versus Fenestration Laminectomy Discectomy for Cervical Disc Herniation.

Objective: It aimed to investigate the difference in clinical efficacy between posterior cervical endoscopic discectomy (PCED) and Fenestration laminectomy discectomy (FLD) in cervical disc herniation (CDH). Methods: This retrospective study analyzed 100 CDH patients undergoing nucleotomy and assigned them into the FLD and PCED groups, 50 cases for each group. The differences in operation time, intraoperative blood loss, skin incision, off-bed time, and hospital stay were evaluated. Numeric rating scales (NRS), Oswestry disability Index (ODI), Japanese Orthopaedic Association (JOA), excellent and good clinical efficacy, quality of life (QoL) SF-36 score, and complication rate were compared. Results: The results showed that compared with the FLD group, the PCED group had increased operation time, decreased intraoperative blood loss, skin incision length, off-bed time, and hospital stay (P < .01). Compared with the FLD group, the PCED group had decreased NRS and ODI scores and increased JOA scores at 1 d, 3 d, 1 month, 3 months, 6 months, 12 months, and 24 months after operation (P < .05). Compared with the FLD group, the excellent and good rate of the PCED group increased significantly after 6 months, 1 year, and 2 years (52.0% vs 64.0%, 58.0% vs. 80.0%, 68.0% vs 90.0%, P < .05). Relative to the FLD group, the physical function, emotional function, vitality, social function, and mental health score of the PCED group increased obviously at 2 years after operation (P < .01). The postoperative complication rate was 0% in both FLD and PCED groups. PCED has good long-term clinical efficacy in the treatment of CDH, with excellent recovery and high safety. Conclusion: PCED showed favorable long-term clinical efficacy in the treatment of CDH, with excellent recovery and high safety. Compared to FLD, PCED resulted in reduced intraoperative blood loss, shorter incision length, and faster recovery. It also led to improved pain scores, functional outcomes, and quality of life measures. The absence of postoperative complications further supports the use of PCED as an effective treatment option for CDH.

Network Pharmacology and Experimental Validation Explore the Pharmacological Mechanisms of Herb Pair for Treating Rheumatoid Arthritis.

OBJECTIVE: This study aimed to elucidate the multitarget mechanism of the Mori Ramulus - Taxilli Herba (MT) herb pair in treating rheumatoid arthritis (RA). METHODS: The targets of the herb pair and RA were predicted from databases and screened through cross-analysis. The core targets were obtained using protein-protein interaction (PPI) network analysis. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed. Finally, animal experiments were conducted to validate the anti-RA effect and mechanism of this herb pair. RESULTS: This approach successfully identified 9 active compounds of MT that interacted with 6 core targets (AKT1, TNF, IL6, TP53, VEGFA, and IL1ß). Pathway and functional enrichment analyses revealed that MT had significant effects on the TNF and IL-17 signaling pathways. The consistency of interactions between active components and targets in these pathways was confirmed through molecular docking. Moreover, the potential therapeutic effect of MT was verified in vivo, demonstrating its ability to effectively relieve inflammation by regulating these targeted genes and pathways. CONCLUSION: The present work suggests that the therapeutic effect of MT herb pair on RA may be attributed to its ability to regulate the TNF signaling pathway and IL-17 signaling pathway.

Initial Stage of Disease Similar for White and Black Patients With Early-Onset Colorectal Cancer at a Safety-Net Hospital.

BACKGROUND: Non-Hispanic Black (NHB) patients with early-onset colorectal cancer (EOCRC) are more likely to present with advanced-stage disease than their Non-Hispanic White (NHW) counterparts. To further elucidate whether differences in tumor biology or disparities in access to care may be responsible, we examined the association between race/ethnicity and initial stage of disease, time to diagnosis, and tumor characteristics among NHW and NHB patients with EOCRC cared for in a safety-net health care setting. METHODS: We performed a retrospective cohort study of NHW and NHB patients diagnosed with primary EOCRC who received care at Boston Medical Center between January 2000 and May 2020. We compared demographics, risk factors, presenting signs/symptoms, time to diagnosis, health care utilization, and tumor characteristics (stage, grade, location, and mutational status). RESULTS: We identified 103 patients (mean age 41.5±7.2 y, 53.4% men), including 40 NHWs and 63 NHBs, with EOCRC. NHB and NHW patients were similar with respect to demographics, presenting signs/symptoms, and risk factor distribution. There were also no significant differences between NHWs and NHBs with respect to the advanced stage of disease at presentation (45.0% vs. 42.9%, P =0.83), the median time to diagnosis [152 d (IQR, 40 to 341) vs. 160 d (IQR, 61 to 312), P =0.79] or tumor characteristics, except for a predilection for proximal disease among NHBs (30.2% vs. 15.0%). CONCLUSIONS: NHB patients were no more likely than NHW patients to present with advanced-stage disease, aggressive tumor histology, or experience delays in diagnosis within a safety-net health care system.

Ultrafine Pt nanoparticles anchored on core-shell structured zeolite-carbon for efficient catalysis of hydrogen generation.

Ammonia borane (AB) is a potential hydrogen storage material with high-efficiency hydrolytic dehydrogenation under a suitable catalyst. Noble metal catalysts have drawn a lot of attention. In this study, a carbon-coated zeolite was obtained by calcination at high temperatures using glucose as a carbon source. Pt nanoparticles were fixed on a core-shell composite support by a simple chemical reduction method. A series of catalysts were prepared with different synthesis parameters. The results show that PSC-2 has excellent catalytic performance for hydrolytic dehydrogenation of AB in alkaline solution at room temperature, and the turnover frequency (TOF) is 593 min-1. The excellent catalytic performance is attributed to the carbon layer on the zeolite surface which inhibits the aggregation or deformation of metals in the catalytic reaction. The metal-support interaction activates the water and accelerates the rate-limiting step of hydrolysis. The activation energy (E a = 44 kJ mol-1) was calculated based on the reaction temperature. In addition, the kinetics of AB hydrolysis was studied, and the effects of catalyst concentration, AB concentration and NaOH concentration on AB hydrolysis rate were further investigated. The high-efficiency catalyst prepared in this work provides a new strategy for the development of chemical hydrogen production in the field of catalysis.

Fabrication of Coral-like Polyaniline/Continuously Reinforced Carbon Nanotube Woven Composite Films for Flexible High-Stability Supercapacitor Electrodes.

The electrochemical performance is significantly influenced by the structure and surface morphology of the electrode materials used in supercapacitors. Using the floating catalytic chemical vapor deposition (FCCVD) technique, a self-supporting, flexible layer of continuously reinforced carbon nanotube woven film (CNWF) was developed. Then, polyaniline (PANI) was formed in the conductive network of CNWF using cyclic voltammetry electrochemical polymerization (CVEP) in various aqueous electrolytes to produce a series of flexible CNWF/PANI composite films. The impacts of the CVEP period, electrolyte type, and electrolyte concentration on the surface morphology, doping degree, and hydrophilicity of CNWF/PANI composite films were thoroughly examined. The CNWF/PANI1-15C composite electrode, which was created using 15 cycles of CVEP in a solution of 1 M sodium bisulfate, displayed a distinctive coral-like PANI layer with a well-defined sharp nanoprotuberance structure, a 48% doping degree, and a quick reversible pseudocapacitive storage mechanism. At a current density of 1 A g-1, the energy density and specific capacitance reached 54.9 Wh kg-1 and 1098.0 F g-1, respectively, with a specific capacitance retention rate of 75.9% maintained at 10 A g-1. Both the specific capacitance and coulomb efficiency were maintained at 96.9% and more than 98.1% of their initial values after being subjected to 2000 cycles of galvanostatic charge and discharge, demonstrating excellent electrochemical cycling stability. The CNWF/PANI1-15C composite film, an ideal electrode material, offers a promising future in the field of flexible energy storage due to its exceptional mechanical properties (127.9 MPa tensile strength and 16.2% elongation at break).

The multi-omics and analysis to reveal thermal processing enhanced anti-rheumatoid arthritis efficacy of Radix Clematidis in rats.

Radix Clematidis (RC) is the roots and rhizomes of Clematis chinensis Osbeck, which has potent effects for expelling wind and dispelling dampness. Processing RC with yellow wine is a traditional processing method. This study aimed to investigate thermal and yellow wine processing influences on potential effective components of RC and its anti-rheumatoid arthritis enhancement mechanisms. Different thermal and wine processing were adopted to get different RC samples. Scanning electron microscope and mercury intrusion porosimetry were used to measure fractal parameters of pore structure. Based on ultra performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS), main constituents were identified and quantified. Besides, the correlation between fractal parameters and main constituents was investigated. The anti-rheumatoid arthritis effect was researched in adjuvant-induced arthritis (AIA) rats. The levels of inflammatory cytokine were determined with ELISA kits. Non-targeted serum metabolomics was performed with UPLC-QTOF/MS. 35 compounds were identified in RC, mainly triterpenoid saponins, also including organic acids and lignanoids. The extraction yield of four active triterpenoid saponins significantly increased because looser tissue and wider pore size distribution. Fractal dimension and total surface area significantly increased while total porosity and total volume decreased. In AIA rats, thermal and wine processed RC could markedly inhibit inflammatory cytokines IL-6, IL-1ß, TNF-α, and VEGF. Besides, tryptophan and lipid metabolism disorders were ameliorated. Thermal and yellow wine treatments engendered complex pore structure to increase the contents of four active triterpenoid saponins of RC, leading to greater anti-rheumatoid arthritis efficacy.

System pharmacology analysis to decipher the effect and mechanism of active ingredients combination from herb couple on rheumatoid arthritis in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Traditional herb couple Angelicae pubescentis radix (APR) and Notopterygii rhizoma et radix (NRR), composition of two traditional Chinese medicinal herbs, has been used clinically in China for the treatment of rheumatoid arthritis (RA) over years. APR and NRR contain coumarins and phenolic acids, which have been reported to have analgesic and anti-inflammatory activities. AIM OF THE STUDY: The active ingredients combination (AIC) and potential therapeutic mechanism of APR and NRR (AN) herb couple remain unclear. Therefore, the present study aimed to identify the AIC and elucidate the underlying mechanism of AIC on RA. MATERIALS AND METHODS: Firstly, a novel strategy of in vitro experiments, computational analysis, UPLC-QTOF-MS and UPLC-QQQ-MS was established to confirm the optimum ratio of AN herb couple samples and identified the AIC. Then, the anti-arthritis effects of the optimal herb couple and AIC were studied with Collagen II induced rheumatoid arthritis (CIA) rats in vivo. Finally, an integrated model of network pharmacology, metabolomics, gut microbiota analysis and biological techniques were applied to clarify the underlying mechanism through a comprehensive perspective. RESULTS: AN7:3 herb couple was regarded as the optimal ratio of AN herbal samples, and AIC was screened as osthole, columbianadin, notopterol, isoimperatorin, psoralen, xanthotoxin, bergapten, nodakenin and bergaptol respectively. Additionally, AIC exerted similar therapeutic effects as AN 7:3 in CIA rats. Moreover, AIC ameliorated RA might via regulating MAPK signaling pathway, altering metabolic disorders and gut microbiome involved autoimmunity. CONCLUSIONS: our findings provided scientific evidence to support that AIC of AN herb couple could be used as a prebiotic agent for RA. Importantly, this research provided a systematic and feasible strategy to optimize the proportion of medicinal materials and screen AIC from multi-component traditional Chinese herb couples or Chinese medicine formulae. Moreover, it provided a comprehensive perspective to discover AIC, clarify the overall effects and understand the mechanisms for natural products through the perspective of database and multi-omics integration.

Prostate cancer identification via photoacoustic spectroscopy and machine learning.

Photoacoustic spectroscopy can generate abundant chemical and physical information about biological tissues. However, this abundance of information makes it difficult to compare these tissues directly. Data mining methods can circumvent this problem. We describe the application of machine-learning methods (including unsupervised hierarchical clustering and supervised classification) to the diagnosis of prostate cancer by photoacoustic spectrum analysis. We focus on the content and distribution of hemoglobin, collagen, and lipids, because these molecules change during the development of prostate cancer. A higher correlation among the ultrasonic power spectra of these chemical components is observed in cancerous than in normal tissues, indicating that the microstructural distributions in cancerous tissues are more consistent. Different classifiers applied in cancer-tissue diagnoses achieved an accuracy of 82 % (better than that of standard clinical methods). The technique thus exhibits great potential for painless early diagnosis of aggressive prostate cancer.

Field carcinogenesis for risk stratification of colorectal cancer.

Colorectal cancer (CRC) is characterized by genetic-environmental interplay leading to diffuse changes in the entire colonic mucosa (field carcinogenesis or field of injury) and to a pro-neoplastic genetic/epigenetic/physiological milieu. The clinical consequences are increased risk of synchronous and metachronous neoplasia. Factors such as genetics, race, ethnicity, age, and socioeconomic status are thought to influence neoplasia development. Here, we explore the potential improvement to CRC screening through exploiting field carcinogenesis, with particular focus on racial disparities and chemoprevention strategies. Also, we discuss future directions for field carcinogenesis/risk stratification using molecular and novel biophotonic techniques for personalized CRC screening.

Impaired prenatal motor axon development necessitates early therapeutic intervention in severe SMA.

Gene replacement and pre-mRNA splicing modifier therapies represent breakthrough gene targeting treatments for the neuromuscular disease spinal muscular atrophy (SMA), but mechanisms underlying variable efficacy of treatment are incompletely understood. Our examination of severe infantile onset human SMA tissues obtained at expedited autopsy revealed persistence of developmentally immature motor neuron axons, many of which are actively degenerating. We identified similar features in a mouse model of severe SMA, in which impaired radial growth and Schwann cell ensheathment of motor axons began during embryogenesis and resulted in reduced acquisition of myelinated axons that impeded motor axon function neonatally. Axons that failed to ensheath degenerated rapidly postnatally, specifically releasing neurofilament light chain protein into the blood. Genetic restoration of survival motor neuron protein (SMN) expression in mouse motor neurons, but not in Schwann cells or muscle, improved SMA motor axon development and maintenance. Treatment with small-molecule SMN2 splice modifiers beginning immediately after birth in mice increased radial growth of the already myelinated axons, but in utero treatment was required to restore axonal growth and associated maturation, prevent subsequent neonatal axon degeneration, and enhance motor axon function. Together, these data reveal a cellular basis for the fulminant neonatal worsening of patients with infantile onset SMA and identify a temporal window for more effective treatment. These findings suggest that minimizing treatment delay is critical to achieve optimal therapeutic efficacy.

Continuously Reinforced Carbon Nanotube Film Sea-Cucumber-like Polyaniline Nanocomposites for Flexible Self-Supporting Energy-Storage Electrode Materials.

The charge storage mechanism and capacity of supercapacitors completely depend on the electrochemical and mechanical properties of electrode materials. Herein, continuously reinforced carbon nanotube film (CNTF), as the flexible support layer and the conductive skeleton, was prepared via the floating catalytic chemical vapor deposition (FCCVD) method. Furthermore, a series of novel flexible self-supporting CNTF/polyaniline (PANI) nanocomposite electrode materials were prepared by cyclic voltammetry electrochemical polymerization (CVEP), with aniline and mixed-acid-treated CNTF film. By controlling the different polymerization cycles, it was found that the growth model, morphology, apparent color, and loading amount of the PANI on the CNTF surface were different. The CNTF/PANI-15C composite electrode, prepared by 15 cycles of electrochemical polymerization, has a unique surface, with a "sea-cucumber-like" 3D nanoprotrusion structure and microporous channels formed via the stacking of the PANI nanowires. A CNTF/PANI-15C flexible electrode exhibited the highest specific capacitance, 903.6 F/g, and the highest energy density, 45.2 Wh/kg, at the current density of 1 A/g and the voltage window of 0 to 0.6 V. It could maintain 73.9% of the initial value at a high current density of 10 A/g. The excellent electrochemical cycle and structural stabilities were confirmed on the condition of the higher capacitance retention of 95.1% after 2000 cycles of galvanostatic charge/discharge, and on the almost unchanged electrochemical performances after 500 cycles of bending. The tensile strength of the composite electrode was 124.5 MPa, and the elongation at break was 18.9%.

2-Deoxy-d-glucose Promotes Buforin IIb-Induced Cytotoxicity in Prostate Cancer DU145 Cells and Xenograft Tumors.

Inhibition of the glycolytic pathway is a critical strategy in anticancer therapy because of the role of aerobic glycolysis in cancer cells. The glycolytic inhibitor 2-Deoxy-d-glucose (2-DG) has shown potential in combination with other anticancer agents. Buforin IIb is an effective antimicrobial peptide (AMP) with broad-spectrum anticancer activity and selectivity. The efficacy of combination treatment with 2-DG and buforin IIb in prostate cancer remains unknown. Here, we tested the efficacy of buforin IIb as a mitochondria-targeting AMP in the androgen-independent human prostate cancer cell line DU145. Combining 2-DG with buforin IIb had a synergistic toxic effect on DU145 cells and mouse xenograft tumors. Combination treatment with 2-DG and buforin IIb caused stronger proliferation inhibition, greater G1 cell cycle arrest, and higher apoptosis than either treatment alone. Combination treatment dramatically decreased L-lactate production and intracellular ATP levels, indicating severe inhibition of glycolysis and ATP production. Flow cytometry and confocal laser scanning microscopy results indicate that 2-DG may increase buforin IIb uptake by DU145 cells, thereby increasing the mitochondria-targeting capacity of buforin IIb. This may partly explain the effect of combination treatment on enhancing buforin IIb-induced apoptosis. Consistently, 2-DG increased mitochondrial dysfunction and upregulated Bax/Bcl-2, promoting cytochrome c release to initiate procaspase 3 cleavage induced by buforin IIb. These results suggest that 2-DG sensitizes prostate cancer DU145 cells to buforin IIb. Moreover, combination treatment caused minimal hemolysis and cytotoxicity to normal WPMY-1 cells. Collectively, the current study demonstrates that dual targeting of glycolysis and mitochondria by 2-DG and buforin IIb may be an effective anticancer strategy for the treatment of some advanced prostate cancer.

Antiproliferative activities of tricyclic amides derived from ß-caryophyllene via the Ritter reaction against MDA-MB-231 breast cancer cells.

A library of novel tricyclic amides has been synthesised via the Ritter reaction from ß-caryophyllene 1 and its monoepoxy derivative 4. The compounds were assessed for antiproliferative activities against the aggressive MDA-MB-231 breast cancer cell line. Of the synthesised compounds, eight were active. 3c and 6b were the most potent and inhibited proliferation with IC50 of 9.7 and 8.2 µM, respectively. Mechanistic studies revealed differences in their antiproliferative actions. 6b inhibited cell cycle progression and induced predominantly apoptotic cell death. In contrast, 3c did not affect cell cycle kinetics and favoured necrotic over apoptotic pathways. Screening against mammalian cells (VERO cells) indicates that 3c and 6b were more active towards MDA-MB-231 cells than noncancerous cells. Facile synthesis and biological results suggest that these caryophyllene derived amides are viable lead compounds for further development.

Novel peptide myristoly-CM4 induces selective cytotoxicity in leukemia K562/MDR and Jurkat cells by necrosis and/or apoptosis pathway.

Purpose: There is an urgent need for the development of novel, effective, and less toxic drugs to treat leukemia. Antimicrobial peptides (AMPs) have received much more attention as alternative chemotherapeutic agents. This study aimed to examined the cytotoxicity of a novel AMP myristoly-CM4 against chronic myeloid leukemia cells (K562/MDR) and acute lymphocytic leukemia cells (Jurkat), and further investigated its selectivity to clarify the cytotoxic mechanism. Materials and methods: In this study, the cytotoxicity and selectivity of myristoly-CM4 against K562/MDR and Jurkat cells were assessed in vitro, and the anticancer mechanism responsible for its cytotoxicity and selectivity was further investigated. Results: Myristoly-CM4 was cytotoxic to these leukemia cell lines (IC50 2-4 µM) and was less cytotoxic to normal cells (HEK-293, L02 cells, peripheral blood mononuclear cells, and erythrocytes). Myristoyl-CM4 had stronger affinity to K562/MDR and Jurkat cells than to normal cells, while the contents of phosphatidylserine and sialic acids on the cell surfaces of K562/MDR and Jurkat cells were significantly higher than that of HEK293 cells. The myristoyl group effectively mediated the internalization of myristoyl-CM4 to leukemia cells. After internalization, myristoyl-CM4 could target mitochondria and affected mitochondrial function, including disruption of Δψm, increasing the accumulation of ROS, increasing the Bax/Bcl-2 ratio, activating caspase 9 and 3, and PARP to induce mitochondria-dependent apoptosis in both K562/MDR and Jurkat cells. Myristoyl-CM4 also induced K562/MDR cell necrosis by directive membrane disruption, and significantly decreased the level of P-glycoprotein in K562/MDR cells. Conclusion: These results suggested that myristoyl-CM4 showed selective cytotoxicity to leukemia K562/MDR and Jurkat cells by apoptosis and/or necrosis pathway. Myristoyl-CM4, thus, appears to be a promising candidate for leukemia treatment, including multidrug-resistant leukemia.

Label free quantitative analysis of Alicyclobacillus acidoterrestris spore germination subjected to low ambient pH.

Inhibition of spore germination or sterilization after induction of spore germination would effectively control low pH food spoilage caused by Alicyclobacillus acidoterrestris spores. However, the characteristics and mechanisms of A. acidoterrestris spore germination in low ambient pH remains poorly understood. In this study, the germination rate of A. acidoterrestris spores at different ambient pH conditions was determined, and subsequently the proteomic profiles of A. acidoterrestris in spore germination were analysed by label-free quantification, in which the specific metabolic pathways involved were identified and key functional proteins were screened and validated using RT-qPCR (real time quantitative PCR). The suitable ambient pH value for the spore germination of A. acidoterrestris ranged from 3.0 to 5.0 with the optimum pH of 4.0. According to the LC-ESI-MS/MS (liquid chromatography electrospray ionization tandem mass spectrometry) analysis, 98 proteins of geminated spores of A. acidoterrestris incubated for 2 h at pH 3.0 were changed significantly in comparison to non-germinated spores, the expression of 20 proteins were up-regulated and that of 78 proteins down-regulated respectively. Those differential expressed proteins were mainly involved in cell wall hydrolysis, cell morphological changes, protein synthesis and folding, perception of external stimuli and signal transduction etc., and we observed that germination receptor D (GerD), cell wall hydrolase, transpeptidase, peptidase S1 and two-component regulatory system phoR were significantly up-regulated, but hydrolase NlpC/P60, peptidoglycan glycosyltransferase, spore coat proteins CotX, CotJB and the Lrp/AsnC (leucine-responsive regulatory protein/asparagine synthase C products) protein were significantly down-regulated in the experiment, which implied the important roles of identified proteins during the spore germination. Furthermore, the pathway analysis showed the possible involvement of differentially expressed proteins in the ß-lactam resistance, ribosome, biosynthesis of secondary metabolites, pyruvate metabolism, two-component system and other metabolic pathways, which indicated that synthesis and hydrolysis of cell wall, intracellular substance synthesis, energy generation and signal transduction were likely associated with the initiation of spore germination and restoration of vegetative growth. In conclusion, the quantitative proteomic landscape of A. acidoterrestris spores could provide the theoretic and experimental evidences for the hazard control of A. acidoterrestris spores in the thermal pasteurization process of acidic beverages industry.

N-myristoylation of Antimicrobial Peptide CM4 Enhances Its Anticancer Activity by Interacting With Cell Membrane and Targeting Mitochondria in Breast Cancer Cells.

Development of antimicrobial peptides (AMPs) as highly effective and selective anticancer agents would represent great progress in cancer treatment. Here we show that myristoyl-CM4, a new synthetic analog generated by N-myristoylation of AMPs CM4, had anticancer activity against MCF-7, MDA-MB-231, MX-1 breast cancer cells (IC50 of 3-6 µM) and MDA-MB-231 xenograft tumors. The improved activity was attributed to the effect of myristoyl on the cell membrane. Flow cytometry and confocal laser scanning microscopy results showed that N-myristoylation significantly increased the membrane affinity toward breast cancer cells and also effectively mediated cellular entry. Despite increasing cytotoxicity against HEK293 and NIH3T3 cells and erythrocytes associated with its anticancer activity, myristoyl-CM4 maintained a certain selectivity toward breast cancer cells. Accordingly, the membrane affinity toward breast cancer cells was two to threefold higher than that of normal cells. Glycosylation analysis showed that sialic acid-containing oligosaccharides (including O-mucin and gangliosides) were important targets for myristoyl-CM4 binding to breast cancer cells. After internalization, co-localization analysis revealed that myristoyl-CM4 targeted mitochondria and induced mitochondrial dysfunction, including alterations in mitochondrial transmembrane potential, reactive oxygen species (ROS) generation and cytochrome c release. Activation of caspase 9, caspase 3 and cleavage of PARP were observed in MX-1, MCF-7, and MDA-MB-231 cells after myristoyl-CM4 treatment. The current work indicates that increasing hydrophobicity by myristoylation to modulate peptide-membrane interactions and then target mitochondria is a good strategy to develop AMPs as anticancer agents in the future.

A network pharmacology approach to determine the synergetic mechanisms of herb couple for treating rheumatic arthritis.

PURPOSE: The purpose of this study was to investigate the therapeutic mechanism(s) of Clematis chinensis Osbeck/Notopterygium incisum K.C. Ting ex H.T (CN). METHODS: A network pharmacology approach integrating prediction of ingredients, target exploration, network construction, module partition and pathway analysis was used. RESULTS: This approach successfully helped to identify 12 active ingredients of CN, interacting with 13 key targets (Akt1, STAT3, TNFsf13, TP53, EPHB2, IL-10, IL-6, TNF, MAPK8, IL-8, RELA, ROS1 and STAT4). Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis indicated that CN-regulated pathways were mainly classified into signal transduction and immune system. CONCLUSION: The present work may help to illustrate the mechanism(s) of action of CN, and it may provide a better understanding of antirheumatic effects.

Molecular and biological characterization of gamma-interferon-inducible lysosomal thiol reductase in silver carp (Hypophthalmichthys molitrix).

Gamma-interferon-inducible lysosomal thiol reductase (GILT) plays an important role in the processing of major histocompatibility complex (MHC) class II-restricted antigens by catalyzing disulfide bonds reduction. Herein, a GILT homolog (ScGILT) was identified from silver carp. Its open reading frame covers 771 base pairs, encoding a protein of 256 amino acids that possesses GILT signature sequence CQHGX2ECX2NX4C, active-site CXXC motif, and two potential N-linked glycosylation sites. The predicted tertiary structures of ScGILT and other GILTs were quite similar in shape and positional arrangement of the key motifs. ScGILT mRNA was constitutively expressed in all detected tissues, with high-level expression in fish immune organs, spleen and head kidney. After stimulation with lipopolysaccharide, the expression of ScGILT mRNA significantly increased in spleen and head kidney cells, and ScGILT protein translocated to late endosomes and lysosomes in HeLa cells. Recombinant ScGILT fused with a His6 tag was expressed and purified, and could reduce the interchain disulfide bonds of IgG at pH 4.5. These results suggested that ScGILT was capable of catalyzing disulfide bonds reduction, and then might play an important role in the processing of MHC class II-restricted antigens in silver carp.