Publications by authors named "Fei Fang"

120 Publications

Fluid Shear Stress Activates YAP to Promote Epithelial-mesenchymal Transition in Hepatocellular Carcinoma.

Mol Oncol 2021 Jul 14. Epub 2021 Jul 14.

Institute of Biomedical Engineering, West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, Chengdu, 610041, China.

Epithelial-mesenchymal transition (EMT) mediated by fluid shear stress (FSS) in the tumor microenvironment plays an important role in driving metastasis of the malignant tumor. As a mechanotransducer, Yes-associated protein (YAP) is known to translocate into the nucleus to initiate transcription of genes involved in cell proliferation upon extracellular biophysical stimuli. Here, we showed that FSS facilitated cytoskeleton rearrangement in hepatocellular carcinoma cells, which led to the release of YAP from its binding partner, integrin β subunit, in the cytomembrane. Moreover, we found that upregulation of GEF-H1, a microtubule-associated Rho guanine nucleotide exchange factor (GEF), is a critical step in the FSS-induced translocation of YAP. Nuclear YAP activated the expression of the EMT-regulating transcription factor SNAI1, but suppressed the expression of N6-methyladenosine (m A) modulators; together, this promoted the expression of EMT-related genes. We also observed that FSS-treated HepG2 cells showed markedly increased tumorigenesis and metastasis in vivo. Collectively, our findings unravel the underlying molecular processes by which FSS induces translocation of YAP from the cytomembrane to the nucleus, contributes to EMT and enhances metastasis in hepatocellular carcinoma.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/1878-0261.13061DOI Listing
July 2021

The Stability of Diphosphino-Boryl PBP Pincer Backbone: PBP to POP Ligand Hydrolysis.

Chem Asian J 2021 Jul 12. Epub 2021 Jul 12.

Henan Key Laboratory of Boron Chemistry and Advanced Energy Materials, Key Laboratory of Green Chemical Media and Reactions, Ministry of Education, Collaborative Innovation Centre of Henan Province for Green Manufacturing of Fine Chemicals, School of Chemistry and Chemical Engineering, Henan Normal University, Xinxiang, Henan, 453007, P. R. China.

Since moisture may frequently be present in many solvents, it is important to know the reactivity of a catalyst against water for catalytic reactions. In order to explore the stability and understand the transformation process of diphosphino-boryl-based PBP pincer platform, [PdCl{B(NCH P Bu ) -o-C H }] (1) was treated with PdCl , HB(NCH PPh ) -o-C H was reacted with [PdCl (cod)] (cod=cyclo-octa-1,5-diene) and [Pd (dba) ] (dba=dibenzylideneacetone), respectively, in the presence of water. Some novel palladium POP complexes, [Pd Cl (μ-Cl){μ-κ -P,O,P-OB(NCH P Bu ) -o-C H }] (2 a), [Pd (μ-Cl) (μ-O) {μ-κ -P,O,P-OB(NCH PPh ) -o-C H } ] (2 b), [Pd {μ-κ -P,P,P,P-O(B(NCH PPh ) -o-C H ) }{μ-κ -P,P-(NHCH PPh ) -o-C H }] (3), were obtained. It was found that the PBP pincer backbone can easily be converted into a POP backbone in the presence of water. From the crystal structures of the resultant palladium complexes, possible pincer backbone transformation pathways were discussed.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/asia.202100690DOI Listing
July 2021

Selective Molecular Recognition of Low Density Lipoprotein Based on β-Cyclodextrin Coated Electrochemical Biosensor.

Biosensors (Basel) 2021 Jun 30;11(7). Epub 2021 Jun 30.

MOE Key Laboratory of Macromolecular Synthesis and Functionalization, Department of Polymer Science and Engineering, Zhejiang University, Hangzhou 310027, China.

The excess of low-density lipoprotein (LDL) strongly promotes the accumulation of cholesterol on the arterial wall, which can easily lead to the atherosclerotic cardiovascular diseases (ACDs). It is a challenge on how to recognize and quantify the LDL with a simple and sensitive analytical technology. Herein, β-cyclodextrins (β-CDs), acting as molecular receptors, can bind with LDL to form stable inclusion complexes via the multiple interactions, including electrostatic, van der Waals forces, hydrogen bonding and hydrophobic interactions. With the combination of gold nanoparticles (Au NPs) and β-CDs, we developed an electrochemical sensor providing an excellent molecular recognition and sensing performance towards LDL detection. The LDL dynamic adsorption behavior on the surface of the β-CD-Au electrode was explored by electrochemical impedance spectroscopy (EIS), displaying that the electron-transfer resistance (Ret) values were proportional to the LDL (positively charged apolipoprotein B-100) concentrations. The β-CD-Au modified sensor exhibited a high selectivity and sensitivity (978 kΩ·µM) toward LDL, especially in ultra-low concentrations compared with the common interferers HDL and HSA. Due to its excellent molecular recognition performance, β-CD-Au can be used as a sensing material to monitor LDL in human blood for preventing ACDs in the future.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3390/bios11070216DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8301920PMC
June 2021

The role of loading in murine models of rotator cuff disease.

J Orthop Res 2021 Jun 3. Epub 2021 Jun 3.

Department of Orthopaedic Surgery, Columbia University Irving Medical Center, New York, New York, USA.

Rotator cuff disease pathogenesis is associated with intrinsic (e.g., age, joint laxity, muscle weakness) and extrinsic (e.g., mechanical load, fatigue) factors that lead to chronic degeneration of the cuff tissues. However, etiological studies are difficult to perform in patients due to the long duration of disease onset and progression. Therefore, the purpose of this study was to determine the effects of altered joint loading on the rotator cuff. Mice were subjected to one of three load-dependent rotator cuff tendinopathy models: underuse loading, achieved by injecting botulinum toxin-A into the supraspinatus muscle; overuse loading, achieved using downhill treadmill running; destabilization loading, achieved by surgical excision of the infraspinatus tendon. All models were compared to cage activity animals. Whole joint function was assessed longitudinally using gait analysis. Tissue-scale structure and function were determined using microCT, tensile testing, and histology. The molecular response of the supraspinatus tendon and enthesis was determined by measuring the expression of 84 wound healing-associated genes. Underuse and destabilization altered forepaw weight-bearing, decreased tendon-to-bone attachment strength, decreased mineral density of the humeral epiphysis, and reduced tendon strength. Transcriptional activity of the underuse group returned to baseline levels by 4 weeks, while destabilization had significant upregulation of inflammation, growth factors, and extracellular matrix remodeling genes. Surprisingly, overuse activity caused changes in walking patterns, increased tendon stiffness, and primarily suppressed expression of wound healing-related genes. In summary, the tendinopathy models demonstrated how divergent muscle loading can result in clinically relevant alterations in rotator cuff structure, function, and gene expression.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/jor.25113DOI Listing
June 2021

New insight into the formation mechanism of 2-furfurylthiol in the glucose-cysteine reaction with ribose.

Food Res Int 2021 05 16;143:110295. Epub 2021 Mar 16.

Institute of Food Science and Technology, Chinese Academy of Agricultural Sciences, Key Laboratory of Agro-Products Processing, Ministry of Agriculture and Rural Affairs, Beijing 100193, China. Electronic address:

The molecular formation mechanism of 2-furfurylthiol in the glucose-cysteine reaction is not reported. Knowledge of the molecular interaction of glucose and ribose on the generation of 2-furfurylthiol is still unclear. The carbon module labeling (CAMOLA) technical approach was applied to elucidate the formation mechanism of 2-furfurylthiol in the glucose-cysteine reaction. The effect of ribose on the glucose-cysteine reaction was also evaluated. The results showed that 2-furfural and 2-furanmethanol were important intermediates for the formation of 2-furfurylthiol. Irrespective of the heating time, 2-furfurylthiol was mainly generated from an intact C glucose skeleton (88-89%), whereas the recombination of glucose fragments had minimal contribution. 2-Furfural could be generated from the Maillard reaction between glucose and cysteine or glucose alone, which further formed 2-furanmethanol. Immediately, 2-furfurylthiol could arise from the reaction of 2-furanmethanol and HS from cysteine. Moreover, the reaction of glucose, ribose, and cysteine could generate 2-furfural, 2-furanmethanol, and 2-furfurylthiol by an addition effect confirmed by the model reaction and food system.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.foodres.2021.110295DOI Listing
May 2021

CN-3 induces mitochondrial apoptosis in glioma via Ros-mediated PI3K/AKT pathway.

Pharmazie 2021 May;76(5):208-214

Department of Neurosurgery, Xijing Institute of Clinical Neuroscience, Air Force Medical University, Xi'an, P. R. China;, Email:

Recently we isolated CN-3, a new asterosaponin from starfish , and reported that asterosaponin arrests glioma cell cycle via SCUBE3. However, the multiple mechanisms underlying CN-3 anti-glioma action remains poorly known. Thus, the focus of this study was to evaluate the inhibitory effect of CN-3 on human glioma cells and its underlying molecular mechanisms. U87 and U251 cells were incubated with various concentrations of CN-3, and CCK-8, transmission electron microscopy, ICELLigence, TUNEL, flow cytometry, -acetyl-L-cysteine, and western blot were conducted. As a result, it was found that CN-3 significantly inhibited U87 and U251 cell viability and proliferation in a time- and dose- dependent manner, and also induced mitochondrial apoptosis. Furthermore, we detected that CN-3 downregulated PI3K, P-Akt, AKT and BCL-2, and upregulated cytochrome C and BAX in U87 and U251 cells. Moreover, ROS triggered the inhibition and cell apoptosis for CN-3 via inactivation of P-Akt and activation of cytochrome C. In conclusion, these findings suggest that CN-3 may be a promising candidate for the development of a therapy of glioma.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1691/ph.2021.1363DOI Listing
May 2021

[Expression of cyclophilin A in oral squamous cell carcinoma and its effect on cell proliferation and invasion].

Hua Xi Kou Qiang Yi Xue Za Zhi 2021 Apr;39(2):164-169

Dept. of Stomatology, Wuhan University People,s Hospital, Wuhan 430060, China.

Objectives: To investigate the expression of cyclophilin A (CyPA) in oral squamous cell carcinoma (OSCC) and explore the effect of downregulating the expression of CyPA gene on the proliferation and invasion of SCC-25 cells.

Methods: A total of 77 cases of patients with OSCC were selected. The expression levels of CyPA proteins in OSCC and adjacent normal tissues were evaluated. SCC-25 cells were cultured and divided into the CyPA interference sequence group, negative control group, and blank group. The expression levels of CyPA mRNA and protein in cells were detected by using real-time fluorescent quantitative polymerase chain reaction and Western blot, respectively. Cell proliferation was detected by using methyl thiazolyl tetrazolium and plate colony formation assays. Cell invasion was detected by using Transwell assay.

Results: The positive expression rate of CyPA protein in OSCC tissues was 76.62%, which was higher than that in adjacent tissues (<0.05). The positive expression rate of CyPA protein in TNM stage T+T, clinical stage Ⅲ+Ⅳ, moderately or poorly differentiated lymph node metastasis was increased (<0.05). Compared with the negative control and blank groups, the CyPA interference sequence group had decreased relative expression levels of CyPA mRNA and protein (<0.05); optical density va-lues of cells at 24, 48, 72, and 96 h (<0.05); and number of cell colonies and invasive cells (<0.05).

Conclusions: The CyPA protein is highly expressed in OSCC tissues, and the downregulation of CyPA gene expression in SCC-25 cells can reduce cell proliferation and inhibit cell invasion.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.7518/hxkq.2021.02.006DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8055773PMC
April 2021

SOX13 regulates cancer stem-like properties and tumorigenicity in hepatocellular carcinoma cells.

Am J Cancer Res 2021 1;11(3):760-772. Epub 2021 Mar 1.

Fujian Provincial Key Laboratory of Chronic Liver Disease and Hepatocellular Carcinoma, Xiamen Translational Medical Key Laboratory of Digestive Tumor, Zhongshan Hospital, Xiamen University Xiamen 361004, People's Republic of China.

Sex-determining region Y (SRY)-related high mobility group (HMG) box (SOX) proteins are pivotal transcriptional factors that play essential roles in embryonic development, cell fate decisions and cancer development. The molecular mechanism of SOX13, a member of the SOX family, in hepatocellular carcinoma (HCC) remains largely unknown. In the current study, we found that HCC cells were able to form spheroids in serum-free suspension culture and that SOX13 expression was upregulated in spheroids enriched for cancer stem cells (CSCs). Inhibition of SOX13 in HCC-LM3 and MHCC-97H cells decreased the expression of stemness-related genes; attenuated spheroid formation, anchor-dependent and anchor-independent cell proliferation and tumorigenicity; and enhanced sensitivity to drug treatment. Furthermore, based on analysis of TCGA dataset, the results indicated that SOX13 expression was obviously upregulated and closely associated with poor prognosis in HCC patients. Moreover, SOX13 was correlated with TAZ and CD24 expression. These data strongly demonstrated that SOX13 is involved in maintaining cancer stem-like properties in HCC cells and plays a critical role in HCC development.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7994154PMC
March 2021

The Combined Contribution of Vascular Endothelial Cell Migration and Adhesion to Stent Re-endothelialization.

Front Cell Dev Biol 2021 4;9:641382. Epub 2021 Mar 4.

West China School of Basic Medical Sciences and Forensic Medicine, Institute of Biomedical Engineering, Sichuan University, Chengdu, China.

Coronary stent placement inevitably causes mechanical damage to the endothelium, leading to endothelial denudation and in-stent restenosis (ISR). Re-endothelialization depends mainly on the migration of vascular endothelial cells (VECs) adjacent to the damaged intima, as well as the mobilization and adhesion of circulating VECs. To evaluate the combined contribution of VEC migration and adhesion to re-endothelialization under flow and the influence of stent, models were constructed to simulate various endothelial denudation scales (2 mm/5 mm/10 mm) and stent deployment depths (flat/groove/bulge). Our results showed that (1) in 2 mm flat/groove/bulge models, both VEC migration and adhesion combined completed the percentage of endothelial recovery about 27, 16, and 12%, and migration accounted for about 21, 15, and 7%, respectively. It was suggested that the flat and groove models were in favor of VEC migration. (2) With the augmentation of the injury scales (5 and 10 mm), the contribution of circulating VEC adhesion on endothelial repair increased. Taken together, endothelial restoration mainly depended on the migration of adjacent VECs when the injury scale was 2 mm. The adhered cells contributed to re-endothelialization in an injury scale-dependent way. This study is helpful to provide new enlightenment for surface modification of cardiovascular implants.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3389/fcell.2021.641382DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7969796PMC
March 2021

Therapeutic effect of intense pulsed light with optimal pulse technology on meibomian gland dysfunction with and without ocular infestation.

Ann Transl Med 2021 Feb;9(3):238

Department of Ophthalmology, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, China.

Background: To evaluate the therapeutic effect of intense pulsed light (IPL) therapy with optimal pulse technology (OPT) on meibomian gland dysfunction (MGD) with and without ocular infestation.

Methods: This prospective study included 150 patients with MGD who were divided into -infested (DI) and non-infested (control) groups according to counts determined by epilating eyelashes to evaluate. Each patient underwent three OPT treatment sessions at 3-week intervals. Best corrected visual acuity (BCVA), intraocular pressure (IOP), counts, Ocular Surface Disease Index (OSDI) score, conjunctival congestion, tear meniscus height (TMH), tear breakup time (TBUT), Schirmer I test (SIT), corneal and conjunctival fluorescein staining scores, meibomian gland (MG) macrostructure, lid margin abnormality, MG expressibility, and meibum quality were assessed before the first treatment as baseline and at 1, 2, and 3 months (M1, M2 and M3, respectively) after treatment.

Results: The incidence of infestation was 59.15% (84/142) at baseline and the final eradication rate at M3 was 83.3% (70/84). Corneal staining and conjunctival congestion showed slower improvement in the DI group than in the control group at M1 and M2 (P<0.05), but was significantly improved at M3 than that at with baseline (P<0.01). Both the groups showed significant improvement in OSDI, conjunctival congestion, TBUT, corneal and conjunctival staining, MG dropout, lid margin abnormality, MG expressibility, and meibum quality at M3 (P<0.05). No differences were observed for BCVA, IOP, SIT, and TMH before and after treatment in both the groups (P>0.05).

Conclusions: This new-generation IPL with OPT treatment results in greater improvement in MG expressibility and meibum quality in MGD patients with infestation than in those without, but not for corneal staining.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.21037/atm-20-1745DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7940958PMC
February 2021

RING-finger protein 5 promotes hepatocellular carcinoma progression and predicts poor prognosis.

Hum Cell 2021 Mar 19;34(2):530-538. Epub 2021 Jan 19.

School of Clinical Medicine, Fujian Medical University, Fuzhou, China.

Hepatocellular carcinoma (HCC) is one of the most common cause of malignancy-related deaths. Recently, RING-finger protein 5 (RNF5), an E3 ubiquitin ligase, was revealed to be associated with the development of several human cancers. However, the clinical implication and functional role of RNF5 in HCC are poorly understood. We analysed RNF5 expression in HCC samples and observed that both the mRNA and protein levels of RNF5 were significantly increased in HCC tissues. RNF5 upregulation was markedly associated with larger tumour size, more satellite foci, and higher alpha fetoprotein (AFP) level, indicating poor prognosis in patients with HCC. Knockdown and overexpression experiments demonstrated that RNF5 promoted the proliferation, migration, and invasion of HCC cells in vitro. Moreover, RNF5 facilitated HCC growth in vivo. Our findings indicated that RNF5 was an oncogene of HCC progression and could be used as a novel prognostic biomarker and therapeutic target for patients with HCC.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s13577-020-00460-5DOI Listing
March 2021

Aligned Biofunctional Electrospun PLGA-LysoGM1 Scaffold for Traumatic Brain Injury Repair.

ACS Biomater Sci Eng 2020 04 4;6(4):2209-2218. Epub 2020 Mar 4.

School of Medicine, Chongqing University, Chongqing 400044, China.

Due to poor regenerative capabilities of the brain, a treatment for traumatic brain injury (TBI) presents a serious challenge to modern medicine. Biofunctional scaffolds that can support neuronal growth, guide neurite elongation, and re-establish impaired brain tissues are urgently needed. To this end, we developed an aligned biofunctional scaffold (aPLGA-LysoGM1), in which poly (lactic--glycolic acid) (PLGA) was functionalized with sphingolipid ceramide -deacylase (SCDase)-hydrolyzed monosialotetrahexosylganglioside (LysoGM1) and electrospinning was used to form an aligned fibrous network. As a ganglioside of neuronal membranes, the functionalized LysoGM1 endows the scaffold with unique biological properties favoring the growth of neuron and regeneration of injured brain tissues. Moreover, we found that the aligned PLGA-LysoGM1 fibers acted as a topographical cue to guide neurite extension, which is critical for organizing the formation of synaptic networks (neural networks). Systematic in vitro studies demonstrated that the aligned biofunctional scaffold promotes neuronal viability, neurite outgrowth, and synapse formation and also protects neurons from pressure-related injury. Additionally, in a rat TBI model, we demonstrated that the implantation of aPLGA-LysoGM1 scaffold supported recovery from brain injury, as more endogenous neurons were found to migrate and infiltrate into the defect zone compared with alternative scaffold. These results suggest that the aligned biofunctional aPLGA-LysoGM1 scaffold represents a promising therapeutic strategy for brain tissue regeneration following TBI.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1021/acsbiomaterials.9b01636DOI Listing
April 2020

Efficient urine removal, simultaneous elimination of emerging contaminants, and control of toxic chlorate in a photoelectrocatalytic-chlorine system.

Environ Pollut 2020 Dec 8;267:115605. Epub 2020 Sep 8.

School of Environmental Science and Engineering, Shanghai Jiao Tong University, Shanghai, 200240, PR China; Shanghai Institute of Pollution Control and Ecological Security, Shanghai, 200092, PR China; Key Laboratory of Thin Film and Microfabrication Technology, Ministry of Education, Shanghai, 200240, PR China; Yunnan Key Laboratory of Pollution Process and Management of Plateau Lake-Watershed, Yunnan, 650034, PR China. Electronic address:

Urine, which is an important waste biomass resource, is the main source of nitrogen in sewage and contains large quantities of emerging contaminants (ECs). In this study, we propose a new method to efficiently remove urine, simultaneously eliminate ECs, and control the generation of toxic chlorate during urine treatment using a photoelectrocatalytic-chlorine (PEC-Cl) system. A type-II heterojunction of WO/BiVO was used as a photoanode to generate chlorine radicals (Cl•) by decreasing the oxidation potential of WO valence band for the highly selective conversion of urine to N and the simultaneous degradation of ECs in an efficient manner. The method presented surprising results. It was observed that the amount of toxic chlorate was significantly inhibited by circumventing the over-oxidation of Cl by holes or hydroxyl radicals (•OH). Moreover, the removal of urea nitrogen reached 97% within 90 min, while the degradation rate of trimethoprim in urine was above 98.6% within 60 min, which was eight times more than that in the PEC system (12.1%). Compared to the bare WO photoanode, the toxic chlorate and nitrate generated by the WO/BiVO heterojunction photoanode decreased by 61% and 44%, respectively. Thus, this study provides a safe, efficient, and environmentally-friendly approach for the disposal of urine.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.envpol.2020.115605DOI Listing
December 2020

Primary cilia as the nexus of biophysical and hedgehog signaling at the tendon enthesis.

Sci Adv 2020 Oct 30;6(44). Epub 2020 Oct 30.

Department of Orthopedic Surgery, Columbia University, New York, NY, 10032, USA.

The tendon enthesis is a fibrocartilaginous tissue critical for transfer of muscle forces to bone. Enthesis pathologies are common, and surgical repair of tendon to bone is plagued by high failure rates. At the root of these failures is a gap in knowledge of how the tendon enthesis is formed and maintained. We tested the hypothesis that the primary cilium is a hub for transducing biophysical and hedgehog (Hh) signals to regulate tendon enthesis formation and adaptation to loading. Primary cilia were necessary for enthesis development, and cilia assembly was coincident with Hh signaling and enthesis mineralization. Cilia responded inversely to loading; increased loading led to decreased cilia and decreased loading led to increased cilia. Enthesis responses to loading were dependent on Hh signaling through cilia. Results imply a role for tendon enthesis primary cilia as mechanical responders and Hh signal transducers, providing a therapeutic target for tendon enthesis pathologies.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1126/sciadv.abc1799DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7608799PMC
October 2020

Efficient SO Removal and Highly Synergistic HO Production Based on a Novel Dual-Function Photoelectrocatalytic System.

Environ Sci Technol 2020 09 24;54(18):11515-11525. Epub 2020 Aug 24.

School of Environmental Science and Engineering, Shanghai Jiao Tong University, No. 800, Dongchuan Rd, Shanghai 200240, PR China.

The direct conversion of SO to SO is rather difficult for flue gas desulfurization due to its inert dynamic with high reaction activation energy, and the absorption by wet limestone-gypsum also needs the forced oxidation of O to oxidize sulfite to sulfate, which is necessary for additional aeration. Here, we propose a method to remove SO with highly synergistic HO production based on a novel dual-function photoelectrocatalytic (PEC) system in which the jointed spontaneous reaction of desulfurization and HO production was integrated instead of nonspontaneous reaction of O to HO. SO was absorbed by alkali liquor then oxidized quickly into SO by a nanorod α-FeO photoanode, which possessed high alkali corrosion resistance and electron transport properties. HO was produced simultaneously in the cathode chamber on a gas diffusion electrode and was remarkably boosted by the conversion reaction of SO to SO in the anode chamber in which the released chemical energy was effectively used to increase HO. The photocurrent density increased by 40% up to 1.2 mA·cm, and the HO evolution rate achieved 58.8 μmol·L·h·cm with the synergistic treatment of SO, which is about five times than that without SO. This proposed PEC cell system offers a cost-effective and environmental-benign approach for dual purpose of flue gas desulfurization and simultaneous high-valued HO production.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1021/acs.est.0c00886DOI Listing
September 2020

Reduction of SCUBE3 by a new marine-derived asterosaponin leads to arrest of glioma cells in G1/S.

Oncogenesis 2020 Aug 6;9(8):71. Epub 2020 Aug 6.

Department of Neurosurgery, Xijing Institute of Clinical Neuroscience, Air Force Medical University, 710032, Xi'an, People's Republic of China.

Many saponins are characterized as exhibiting a wide spectrum of antitumor activities at low concentrations. Most of the previous studies that aimed to understand the mechanisms underlying anticancer saponins have focused on numerous classical signaling pathways. However, at the oncogene level, little is known about the action of saponins, especially asterosaponin. In this study, CN-3, a new asterosaponin isolated from the starfish Culcita novaeguineae, decreased the proliferation of U87 and U251 cells at low doses in a dose- and time-dependent manner. Microarray analysis revealed CN-3 significantly induced the differential expression of 661 genes that are related to its antiglioma effect in U251. Nine downregulated genes (SCUBE3, PSD4, PGM2L1, ACSL3, PRICKLE1, ABI3BP, STON1, EDIL3, and KCTD12) were selected, for further verification of their low expression. Then, shRNA transfection and high-content screening were performed and significantly decreased U251 cell proliferation rate was only observed for the SCUBE3 knockdown. qPCR confirmed SCUBE3 was highly expressed in U251 and U87 cells, and had medium expression levels in U373 cells. Real-time cellular analysis using iCELLigence demonstrated that SCUBE3 is an oncogene in U251 and U87 cells, with knockdown of SCUBE3 inhibiting U251 and U87 cell proliferation while, conversely, SCUBE3 overexpression promoted their proliferation. Afterward, SCUBE3 protein was found to have high expression in primary glioma specimens from patients examined by immunohistochemistry but low expression in normal brain. PathScan ELISA analysis in conjunction with TEM observation demonstrated that the effect of SCUBE3 knockdown in U251 does not appear to be related to the induction of apoptosis. Employing CCK-8, iCELLigence, flow cytometry, western blotting, and shRNA transfection (knockdown and overexpression) experiments, we reveal that the reduction of SCUBE3 expression, induced by CN-3, mediated both inhibition and G1/S arrest of U251 via the Akt/p-Akt/p53/p21/p27/E2F1 pathway.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1038/s41389-020-00252-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7411020PMC
August 2020

Molecular Dynamics Simulation-assisted Ionic Liquid Screening for Deep Coverage Proteome Analysis.

Mol Cell Proteomics 2020 10 16;19(10):1724-1737. Epub 2020 Jul 16.

CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Science, National Chromatographic Research and Analysis Center, Dalian, China.

In-depth coverage of proteomic analysis could enhance our understanding to the mechanism of the protein functions. Unfortunately, many highly hydrophobic proteins and low-abundance proteins, which play critical roles in signaling networks, are easily lost during sample preparation, mainly attributed to the fact that very few extractants can simultaneously satisfy the requirements on strong solubilizing ability to membrane proteins and good enzyme compatibility. Thus, it is urgent to screen out ideal extractant from the huge compound libraries in a fast and effective way. Herein, by investigating the interior mechanism of extractants on the membrane proteins solubilization and trypsin compatibility, a molecular dynamics simulation system was established as complement to the experimental procedure to narrow down the scope of candidates for proteomics analysis. The simulation data shows that the van der Waals interaction between cation group of ionic liquid and membrane protein is the dominant factor in determining protein solubilization. In combination with the experimental data, 1-dodecyl-3-methylimidazolium chloride (C12Im-Cl) is on the shortlist for the suitable candidates from comprehensive aspects. Inspired by the advantages of C12Im-Cl, an ionic liquid-based filter-aided sample preparation (-FASP) method was developed. Using this strategy, over 3,300 proteins were confidently identified from 10 HeLa cells (∼100 ng proteins) in a single run, an improvement of 53% over the conventional FASP method. Then the -FASP method was further successfully applied to the label-free relative quantitation of human liver cancer and para-carcinoma tissues with obviously improved accuracy, reproducibility and coverage than the commonly used urea-based FASP method. The above results demonstrated that the -FASP method could be performed as a versatile tool for the in-depth coverage proteomic analysis of biological samples.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1074/mcp.TIR119.001827DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8015004PMC
October 2020

Perturbation of Specific Signaling Pathways Is Involved in Initiation of Mouse Liver Fibrosis.

Hepatology 2021 Apr 19;73(4):1551-1569. Epub 2021 Mar 19.

Guangdong Provincial Key Laboratory of Food, Nutrition and Health, Department of Toxicology, School of Public Health, Sun Yat-sen University, Guangzhou, China.

Background And Aims: To identify the regulatory role of protein phosphatase 2A (PP2A) in the development of liver disease, we generated a mouse model with hepatocyte-specific deletion of Ppp2r1a gene (encoding PP2A Aα subunit).

Approach And Results: Homozygote (HO) mice and matched wild-type littermates were investigated at 3, 6, 9, 12, 15, and 18 months of age. Pathological examination showed that PP2A Aα deficiency in hepatocytes resulted in progressive liver fibrosis phenotype from 9 months of age. No hepatocyte death was observed in HO mice. However, perturbation of pathways including epidermal growth factor receptor 1 (EGFR1), amino acid metabolism, and translation factors as well as leptin and adiponectin led to pronounced hepatic fibrosis. In vitro studies demonstrated the involvement of specific B subunit complexes in the regulation of EGFR1 signaling pathway and cross talk between defected hepatocytes and stimulation of interstitial hyperplasia. It is noteworthy that HO mice failed to develop hepatocellular carcinoma for as long as 22 months of age. We further demonstrate that PP2A Aβ-containing holoenzymes played a critical role in preventing hepatocyte apoptosis and antagonizing tumorigenesis through specific pathways on Aα loss. Furthermore, PP2A Aα and Aβ were functionally distinct, and the Aβ isoform failed to substitute for Aα in the development of inflammation and liver fibrosis.

Conclusions: These observations identify pathways that contribute to the pathogenesis of liver fibrosis and provide putative therapeutic targets for its treatment.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/hep.31457DOI Listing
April 2021

Novel 3D Pd-Cu(OH)/CF cathode for rapid reduction of nitrate-N and simultaneous total nitrogen removal from wastewater.

J Hazard Mater 2021 01 29;401:123232. Epub 2020 Jun 29.

School of Environmental Science and Engineering, Shanghai Jiao Tong University, Shanghai 200240, PR China; Shanghai Institute of Pollution Control and Ecological Security, Shanghai 200092, PR China; Key Laboratory of Thin Film and Microfabrication Technology, Ministry of Education, Shanghai 200240, PR China; Yunnan Key Laboratory of Pollution Process and Management of Plateau Lake-Watershed, Yunnan 650034, PR China. Electronic address:

Removal of NO is a challenging problem in wastewater treatment. Electrocatalysis shows a great potential to remove NO but selectively converting NO to N is facing a low efficiency. Here, a novel 3D Pd-Cu(OH)/CF cathode based electrocatalytic (EC) system was proposed that can rapidly and selectively convert NO to NH, and further convert to N simultaneously. The special designs for the system include: Cu(OH) nanowires were firstly grown on copper foam (CF) with excellent conductivity that features high specific surface area in enhancing NO absorption and conversion to NO. Then, palladium (Pd) with a superior photons activation capacity was doped on the Cu(OH) nanowires to promote the reduction of NO to NH. Then NH was quickly oxidized into N by active chlorine. Finally, total nitrogen (TN) could easily be removed completely via above exhaustive cycle reactions. The 3D Pd-Cu(OH)/CF cathode exhibits a 98.8 % conversion of NO to NH in 45 min with the reported highest removal rate of 0.017 cm min, which is 19.4 times higher than that of CF. The converted NH was finally exhaustively oxidized to N with a 98.7 % of TN removal in 60 min.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.jhazmat.2020.123232DOI Listing
January 2021

Allosteric interactions in the parathyroid hormone GPCR-arrestin complex formation.

Nat Chem Biol 2020 10 6;16(10):1096-1104. Epub 2020 Jul 6.

Department of Pharmacology and Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.

Peptide ligands of class B G-protein-coupled receptors act via a two-step binding process, but the essential mechanisms that link their extracellular binding to intracellular receptor-arrestin interactions are not fully understood. Using NMR, crosslinking coupled to mass spectrometry, signaling experiments and computational approaches on the parathyroid hormone (PTH) type 1 receptor (PTHR), we show that initial binding of the PTH C-terminal part constrains the conformation of the flexible PTH N-terminal signaling epitope before a second binding event occurs. A 'hot-spot' PTH residue, His9, that inserts into the PTHR transmembrane domain at this second step allosterically engages receptor-arrestin coupling. A conformational change in PTHR intracellular loop 3 permits favorable interactions with β-arrestin's finger loop. These results unveil structural determinants for PTHR-arrestin complex formation and reveal that the two-step binding mechanism proceeds via cooperative fluctuations between ligand and receptor, which extend to other class B G-protein-coupled receptors.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1038/s41589-020-0567-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7502484PMC
October 2020

Sox13 promotes hepatocellular carcinoma metastasis by transcriptionally activating Twist1.

Lab Invest 2020 11 27;100(11):1400-1410. Epub 2020 May 27.

Fujian Provincial Key Laboratory of Chronic Liver Disease and Hepatocellular Carcinoma, Zhongshan Hospital, Xiamen University, Building 6, No. 209, South Hubin Road, 361004, Xiamen, China.

SRY (sex-determining region Y)-box 13 (Sox13), a member of group D of the SRY-related high mobility group (HMG) box (Sox) family, is a critical regulator of embryonic development and cartilage formation. Few studies have investigated the role of Sox13 in tumorigenesis. The present study reveals the clinical significance and biological function of Sox13 in hepatocellular carcinoma (HCC). First, the expression of Sox13 in HCC samples was evaluated by qRT-PCR and western blotting, and its association with clinicopathological features and prognosis was determined. We found that Sox13 expression was higher in tumor tissue than in paired nontumor tissue. The upregulation of Sox13 was associated with poor differentiation, metastasis, recurrence and poor overall, and tumor-free survival of HCC patients. The function of Sox13 on HCC cell migration and invasion was then assessed by Transwell assay, and the results demonstrated that Sox13 promoted HCC cell invasion, migration, and epithelial-to-mesenchymal transition (EMT). Notably, the invasion, migration, and EMT of HCC cells induced by Sox13 overexpression could be abolished by Twist1 depletion, and Sox13 was positively correlated with Twist1 at both the mRNA and protein levels. Mechanistically, we revealed that Sox13 activated Twist1 transcription and consequently upregulated Twist1 expression. Furthermore, Sox13 formed a heterodimer with Sox5, and this heterodimer functionally cooperated to enhance the transcriptional activity of Twist1. Our findings suggest that Sox13 serves as an oncogene in HCC, and might be a novel prognostic and therapeutic candidate.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1038/s41374-020-0445-0DOI Listing
November 2020

Status, challenges, and future prospects of stem cell therapy in pelvic floor disorders.

World J Clin Cases 2020 Apr;8(8):1400-1413

Department of Obstetrics and Gynecology, Key Laboratory of Birth Defects and Related Diseases of Women and Children of MOE, West China Second University Hospital, West China Campus, Sichuan University, Chengdu 610041, Sichuan Province, China.

Pelvic floor disorders (PFDs) represent a group of common and frequently-occurring diseases that seriously affect the life quality of women, generally including stress urinary incontinence and pelvic organ prolapse. Surgery has been used as a treatment for PFD, but almost 30% of patients require subsequent surgery due to a high incidence of postoperative complications and high recurrence rates. Therefore, investigations of new therapeutic strategies are urgently needed. Stem cells possess strong multi-differentiation, self-renewal, immunomodulation, and angiogenesis abilities and they are able to differentiate into various cell types of pelvic floor tissues and thus provide a potential therapeutic approach for PFD. Recently, various studies using different autologous stem cells have achieved promising results by improving the pelvic ligament and muscle regeneration and conferring the tissue elasticity and strength to the damaged tissue in PFD, as well as reduced inflammatory reactions, collagen deposition, and foreign body reaction. However, with relatively high rates of complications such as bladder stone formation and wound infections, further studies are necessary to investigate the role of stem cells as maintainers of tissue homeostasis and modulators in early interventions including therapies using new stem cell sources, exosomes, and tissue-engineering combined with stem cell-based implants, among others. This review describes the types of stem cells and the possible interaction mechanisms in PFD treatment, with the hope of providing more promising stem cell treatment strategies for PFD in the future.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.12998/wjcc.v8.i8.1400DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7190946PMC
April 2020

Integrative structure and function of the yeast exocyst complex.

Protein Sci 2020 06 1;29(6):1486-1501. Epub 2020 May 1.

Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, California, USA.

Exocyst is an evolutionarily conserved hetero-octameric tethering complex that plays a variety of roles in membrane trafficking, including exocytosis, endocytosis, autophagy, cell polarization, cytokinesis, pathogen invasion, and metastasis. Exocyst serves as a platform for interactions between the Rab, Rho, and Ral small GTPases, SNARE proteins, and Sec1/Munc18 regulators that coordinate spatial and temporal fidelity of membrane fusion. However, its mechanism is poorly described at the molecular level. Here, we determine the molecular architecture of the yeast exocyst complex by an integrative approach, based on a 3D density map from negative-stain electron microscopy (EM) at ~16 Å resolution, 434 disuccinimidyl suberate and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride cross-links from chemical-crosslinking mass spectrometry, and partial atomic models of the eight subunits. The integrative structure is validated by a previously determined cryo-EM structure, cross-links, and distances from in vivo fluorescence microscopy. Our subunit configuration is consistent with the cryo-EM structure, except for Sec5. While not observed in the cryo-EM map, the integrative model localizes the N-terminal half of Sec3 near the Sec6 subunit. Limited proteolysis experiments suggest that the conformation of Exo70 is dynamic, which may have functional implications for SNARE and membrane interactions. This study illustrates how integrative modeling based on varied low-resolution structural data can inform biologically relevant hypotheses, even in the absence of high-resolution data.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/pro.3863DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7255525PMC
June 2020

Ionic liquid-assisted protein extraction method for plant phosphoproteome analysis.

Talanta 2020 Jun 14;213:120848. Epub 2020 Feb 14.

CAS Key Laboratory of Separation Science for Analytical Chemistry, National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Science, Dalian, Liaoning, 116023, China.

Protein phosphorylation is one of the most important post-translational modifications (PTM) and plays critical roles in maintaining many biological processes of plant species, such as being a significant signal related to resistance to tobacco mosaic virus (TMV) infection in tobacco. Compared to other organisms, in-depth profiling of plant phosphoproteome remains challenging due to the harsh extraction environment of plant proteins and low abundance of plant phosphorylation, generally requiring large amount of plant materials. Herein, we developed an integrated strategy for efficient sample preparation of amounts of plant tissues, by integrating ionic liquid (IL)-assisted protein extraction, in-solution digestion, precipitation-assisted IL removal, as well as immobilized metal ion affinity chromatography (IMAC) enrichment of phosphopeptides together. In this strategy, to improve the efficiency of protein extraction and enzymatic digestion, IL of 1-dodecyl-3-methylimidazolium chloride (C12Im-Cl) was used as the solubilizer due to its excellent solubilizing ability and enzyme compatibility demonstrated in our previous work. Briefly, the extraction capability of C12Im-Cl for protein amount from tobacco leaves was improved 1.9-fold compared to the commonly used urea-assisted method. Notably, to avoid its interference with subsequent LC-MS analysis, the IL was easily removed from the peptide solution by our proposed ion substitution-mediated C12Im + precipitation strategy with high efficiency. By handling 10 mg of starting protein materials of tobacco leaves, 14,441 unique phosphopeptides, assigned to 5153 unique phosphoproteins were confidently identified. To the best of our knowledge, this was the most comprehensive phosphorylation dataset for tobacco so far. All the results demonstrated our strategy was of great potential to promote the large-scale analysis of plant phosphoproteome.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.talanta.2020.120848DOI Listing
June 2020

G-dependent regulation of endosomal cAMP generation by parathyroid hormone class B GPCR.

Proc Natl Acad Sci U S A 2020 03 17;117(13):7455-7460. Epub 2020 Mar 17.

Department of Pharmacology and Chemical Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15261;

cAMP production upon activation of G by G protein-coupled receptors has classically been considered to be plasma membrane-delimited, but a shift in this paradigm has occurred in recent years with the identification of several receptors that continue to signal from early endosomes after internalization. The molecular mechanisms regulating this aspect of signaling remain incompletely understood. Here, we investigated the role of G activation by the parathyroid hormone (PTH) type 1 receptor (PTHR) in mediating endosomal cAMP responses. Inhibition of G signaling by FR900359 markedly reduced the duration of PTH-induced cAMP production, and this effect was mimicked in cells lacking endogenous Gα We determined that modulation of cAMP generation by G occurs at the level of the heterotrimeric G protein via liberation of cell surface Gβγ subunits, which, in turn, act in a phosphoinositide-3 kinase-dependent manner to promote the assembly of PTHR-βarrestin-Gβγ signaling complexes that mediate endosomal cAMP responses. These results unveil insights into the spatiotemporal regulation of G-dependent cAMP signaling.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1073/pnas.1918158117DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7132270PMC
March 2020

A steroidal saponin form Paris vietnamensis (Takht.) reverses temozolomide resistance in glioblastoma cells via inducing apoptosis through ROS/PI3K/Akt pathway.

Biosci Trends 2020 May 14;14(2):123-133. Epub 2020 Mar 14.

Institute of Materia Medica, School of Pharmacy, Fourth Military Medical University, Xi'an, China.

Glioblastoma is one of the most difficult cancers to treat with a 5-year overall survival rate less than 5%. Temozolomide (TMZ) is an effective drug for prolonging the overall survival time of patients, while drug-resistance is an important clinical problem at present. Pennogenin-3-α-L-rhamnopyranosyl-(1→4)-[α-Lrhamno-pyranosyl-(1→2)]- β-D-glucopyranoside (N45), a steroidal saponin, was isolated from the rhizomes of Paris vietnamensis (Takht.), which is used as a Traditional Chinese Medicine and has been reported to possess preclinical anticancer efficacy in various cancer types. However, the mechanism of the inhibition of N45 on glioblastoma cells and its possible application in the treatment of chemotherapy-resistant glioblastoma cells are still unknown. In this study, we use cellular methodological experiments including cell counting kit-8 (CCK-8) assay, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining assay, flow cytometry assay, transmission electron microscopy (TEM) and Western blot. The results show that N45 significantly suppresses the proliferation of glioblastoma cells and TMZ-resistant glioblastoma cells (U87R) by inducing mitochondrial apoptosis through reactive oxygen species (ROS)/phosphoinositide 3-kinase (PI3K)/Akt signal pathway, and the N-acetyl-L-cysteine (NAC) combined with N45 effectively reduced N45-mediated apoptosis and reversed the inhibition of PI3K/Akt signal pathway. In addition, N45 decreased the drug-resistance by down-regulation of nuclear factor kappa-B p65 (NF-κB p65) to attenuate O-methylguanine-DNA methyltransferase (MGMT) in TMZ-resistant glioblastoma cells (U87R). Our findings proved that N45 might be a potential therapeutic agent against glioblastoma and TMZ-resistant glioblastoma, promising to be a potential agent to reduce drug resistance.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.5582/bst.2020.01005DOI Listing
May 2020

Quantitative Proteomics for Monitoring Renal Transplant Injury.

Proteomics Clin Appl 2020 07 16;14(4):e1900036. Epub 2020 Feb 16.

Department of Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, PA, 15261, USA.

Purpose: This study is aimed at developing a molecular diagnostics platform to enhance the interpretation of renal allograft biopsies using quantitative proteomic profiling of formalin-fixed and paraffin-embedded (FFPE) specimens.

Experimental Design: A quantitative proteomics platform composed of 1) an optimized FFPE protein sample preparation method, 2) a tandem mass tag TMT10-plex-based proteomic workflow, and 3) a systematic statistical analysis pipeline to reveal differentially expressed proteins has been developed. This platform is then tested on a small sample set (five samples per phenotype) to reveal proteomic signatures that can differentiate T-cell mediated rejection (TCMR) and polyomavirus BK nephropathy (BKPyVN) from healthy functionally stable kidney tissue (STA).

Results: Among 2798 quantified proteins, the expression levels of 740 BKPyVN and 638 TCMR associated proteins are significantly changed compared to STA specimens. Principal component analysis demonstrated good segregation of all three phenotypes investigated. Protein detection and quantitation are highly reproducible: replicate comparative analyses demonstrated 71-84% overlap of detected proteins, and the coefficient of variation for protein measurements is <15% in triplicate liquid chromatography-tandem mass spectrometry runs.

Conclusions And Clinical Relevance: Quantitative proteomics can be applied to archived FFPE specimens to differentiate different causes of renal allograft injury.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/prca.201900036DOI Listing
July 2020

Physicochemical properties and hepatoprotective effects of glycated Snapper fish scale peptides conjugated with xylose via maillard reaction.

Food Chem Toxicol 2020 Mar 3;137:111115. Epub 2020 Jan 3.

College of Biological Science and Technology, Fuzhou University, Fuzhou, Fujian, 350108, China. Electronic address:

The physicochemical properties and hepatoprotective effects of fish scales peptides (FSP) and the glycated peptides conjugated with xylose via Maillard reaction (FSP-MRPs) were investigated. Results showed that the FSP was rich in oligopeptides within 2-10 amino acids, the degree of grafting of FSP-MRPs was 52.97 ± 1.58% and the antioxidant activities in vitro of FSP were improved through Maillard reaction. In order to investigate the antioxidant activities of FSP-MRPs after digestion, the simulated gastrointestinal digestion experiments of FSP and FSP-MRPs in vitro were conducted. Results indicated that the antioxidant activities of FSP and FSP-MRPs remained as stronger as before even under the digestive conditions. Furthermore, FSP-MRPs could significantly reduce the elevated activities of serum aspartate aminotransferase and alanine aminotransferase, decrease the elevated the levels of hepatic malondialdehyde and triglyceride, and inhibit the decrease of hepatic superoxide dismutase, catalase and glutathione peroxidase caused by alcohol-induced liver damage. These findings suggest that the glycated peptides formed by FSP and xylose via Maillard reaction may be potential to be exploited as a potential functional ingredient in food industry.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.fct.2020.111115DOI Listing
March 2020

Smart Cutter: An Efficient Strategy for Increasing the Coverage of Chemical Cross-Linking Analysis.

Anal Chem 2020 01 23;92(1):1097-1105. Epub 2019 Dec 23.

CAS Key Laboratory of Separation Science for Analytical Chemistry, National Chromatographic R. & A. Center , Dalian Institute of Chemical Physics, Chinese Academy of Science , Dalian , Liaoning 116023 , China.

Chemical cross-linking combined with mass spectrometry (CXMS) has emerged as a powerful tool to study protein structure, conformation, and protein-protein interactions (PPIs). Until now, most cross-linked peptides were generated by using commercial cross-linkers, such as DSS, BS, and DSSO, which react with the primary amino groups of the lysine residues of proteins. However, trypsin, the most commonly used proteolytic enzyme, cannot cleave the C-terminus of a linked lysine, making the obtained cross-linked peptides longer than common peptides and unfavorable for MS identification and data searching. Herein, we propose an in situ sequential digestion strategy using enzymes with distinct cleavage specificity, named as smart cutter, to generate cross-linked peptides with suitable length so that the identification coverage could improve. Through the application of such a strategy to DSS cross-linked lysates, additional cross-linked sites (1.3-fold increase) obtained in comparison with those obtained by trypsin-trypsin digestion (2879 vs 1255). Among the different digestion combinations, AspN-trypsin performed the best, with 64% (673/1059) of the cross-linked sites complementary to trypsin-trypsin digestion, which is beneficial to ensure the depth for studying protein structure and PPIs. Taking the 60 kDa chaperonin protein as an example, more than twice the cross-linked sites (30 vs 14) were identified to enrich the protein structure information. In addition, compared to the published protein interaction network for ( http://www.bacteriome.org ), 91 potential PPIs were discovered with our strategy, of which 65 have not covered by trypsin-trypsin digestion. Therefore, these results illustrate the great significance of smart-cutter-based CXMS for the revelation of protein structure as well as finding new PPIs.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1021/acs.analchem.9b04161DOI Listing
January 2020

Activated hepatic stellate cells regulate MDSC migration through the SDF-1/CXCR4 axis in an orthotopic mouse model of hepatocellular carcinoma.

Cancer Immunol Immunother 2019 Dec 23;68(12):1959-1969. Epub 2019 Oct 23.

Fujian Provincial Key Laboratory of Chronic Liver Disease and Hepatocellular Carcinoma, Zhongshan Hospital, Xiamen University, Building 6, No. 209, South Hubin Road, Xiamen, 361004, China.

Hepatic stellate cells (HSCs) are important stromal cells and pivotal mediators involved in the pathogenesis and immunosuppression of hepatocellular carcinoma (HCC). The liver has been demonstrated to be a site for accumulation of tumor-induced myeloid-derived suppressor cells (MDSCs). We previously reported that HSCs induced an increase in the number of MDSCs in HCC. However, how MDSCs are recruited in HCC remains largely unclear. In the present study, we found that HSC-conditioned medium (HSC-CM) induced bone marrow-derived cell and splenocyte migration, especially MDSC migration. Using chemokine-neutralizing antibodies and chemokine receptor inhibitors, we found that HSCs promoted MDSC migration through the SDF-1/CXCR4 axis. Subsequently, we used an orthotopic mouse liver tumor model to determine how HSCs mediated MDSC migration to HCC in vivo. The in vivo results indicated that pretreatment of MDSCs with a CXCR4 inhibitor or injection with SDF-1-knocked down HSCs inhibited MDSC migration to the spleen and liver of the tumor-bearing mice. Together, our findings indicate a central role for HSCs in MDSC migration mediated by the SDF-1/CXCR4 axis, thus revealing a potentially effective approach for modulating the tumor microenvironment by targeting HSCs in HCC.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s00262-019-02414-9DOI Listing
December 2019
-->