Publications by authors named "Jifeng Wang"

85 Publications

LncRNA-encoded microproteins: A new form of cargo in cell culture-derived and circulating extracellular vesicles.

J Extracell Vesicles 2021 Jul 12;10(9):e12123. Epub 2021 Jul 12.

Laboratory of Protein and Peptide Pharmaceuticals and Laboratory of Proteomics Institute of Biophysics Chinese Academy of Sciences Beijing China.

Advancements in omics-based technologies over the past few years have led to the discovery of numerous biologically relevant peptides encoded by small open reading frames (smORFs) embedded in long noncoding RNA (lncRNA) transcripts (referred to as microproteins here) in a variety of species. However, the mechanisms and modes of action that underlie the roles of microproteins have yet to be fully characterized. Herein, we provide the first experimental evidence of abundant microproteins in extracellular vesicles (EVs) derived from glioma cancer cells, indicating that the EV-mediated transfer of microproteins may represent a novel mechanism for intercellular communication. Intriguingly, when examining human plasma, 48, 11 and 3 microproteins were identified from purified EVs, whole plasma and EV-free plasma, respectively, suggesting that circulating microproteins are primarily enriched in EVs. Most importantly, the preliminary data showed that the expression profile of EV microproteins in glioma patient diverged from the health donors, suggesting that the circulating microproteins in EVs might have potential diagnostic application in identifying patients with glioma.
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http://dx.doi.org/10.1002/jev2.12123DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8275822PMC
July 2021

Microbiome-Metabolomics Reveals Endogenous Alterations of Energy Metabolism by the Dushen Tang to Attenuate D-Galactose-Induced Memory Impairment in Rats.

Biomed Res Int 2021 27;2021:6649085. Epub 2021 May 27.

Jilin Provincial Key Laboratory of Biomacromolecules of Chinese Medicine, Jilin Ginseng Academy, Changchun University of Chinese Medicine, Changchun, Jilin 130021, China.

Aging affects the brain function in elderly individuals, and Dushen Tang (DST) is widely used for the treatment of senile diseases. In this study, the protective effect of DST against memory impairment was evaluated through the Morris water maze (MWM) test and transmission electron microscopy (TEM). A joint analysis was also performed using LC-MS metabolomics and the microbiome. The MWM test showed that DST could significantly improve the spatial memory and learning abilities of rats with memory impairment, and the TEM analysis showed that DST could reduce neuronal damage in the hippocampus of rats with memory impairment. Ten potential biomarkers involving pyruvate metabolism, the synthesis and degradation of ketone bodies, and other metabolic pathways were identified by the metabolomic analysis, and it was found that 3-hydroxybutyric acid and lactic acid were involved in the activation of cAMP signaling pathways. The 16S rDNA sequencing results showed that DST could regulate the structure of the gut microbiota in rats with memory impairment, and these effects were manifested as changes in energy metabolism. These findings suggest that DST exerts a good therapeutic effect on rats with memory impairment and that this effect might be mainly achieved by improving energy metabolism. These findings might lead to the potential development of DST as a drug for the treatment of rats with memory impairment.
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http://dx.doi.org/10.1155/2021/6649085DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8175156PMC
May 2021

Deeply Mining a Universe of Peptides Encoded by Long Noncoding RNAs.

Mol Cell Proteomics 2021 Jun 12:100109. Epub 2021 Jun 12.

Laboratory of Protein and Peptide Pharmaceuticals & Laboratory of Proteomics, institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China. Electronic address:

Many small open reading frames (smORFs) embedded in lncRNA transcripts have been shown to encode biologically functional polypeptides (smORFs-encoded polypeptides, SEPs) in different organisms. Despite significant advances in genomics, bioinformatics and proteomics that largely enabled the discovery of novel SEPs, their identification across different biological samples is still hampered by their poor predictability, diminutive size and low relative abundance. Here, we take advantage of NONCODE, a repository containing the most complete collection and annotation of lncRNA transcripts from different species, to build a novel database that attempts to maximize a collection of SEPs from human and mouse lncRNA transcripts. In order to further improve SEP discovery, we implemented two effective and complementary polypeptide enrichment strategies, 30 kDa MWCO filter and C8 SPE column. These combined strategies enabled us to discover 353 and 409 SEPs from, respectively, 8 human cell lines, and 3 mouse cell lines and 8 mouse tissues. Importantly, nineteen of the identified SEPs were then verified through in-vitro expression, immunoblotting, parallel reaction monitoring (PRM) and synthetic peptides. Subsequent bioinformatic analysis revealed that some of the physical and chemical properties of these novel SEPs, including amino acid composition and codon usage, are different from those commonly found in canonical proteins. Intriguingly, nearly 65% of the identified SEPs were found to be initiated with non-AUG start codons. Overall, the strategy presented in this study encompasses an efficient workflow that enabled us to identify 762 novel SEPs across multiple cell lines and tissues, which probably represents the largest number of SEPs detected by mass spectrometry reported to date. These novel SEPs might not only provide new clues for the annotation of noncoding elements in the genome but can also serve as a valuable resource for the functional characterization of individual SEPs.
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http://dx.doi.org/10.1016/j.mcpro.2021.100109DOI Listing
June 2021

Functional Genomic Analyses of the 21q22.3 Locus Identifying Functional Variants and Candidate Gene for Breast Cancer Risk.

Cancers (Basel) 2021 Apr 23;13(9). Epub 2021 Apr 23.

Vanderbilt Epidemiology Center, Vanderbilt-Ingram Cancer Center, Department of Medicine, Division of Epidemiology, Vanderbilt University School of Medicine, Nashville, TN 37203, USA.

We previously identified a locus at 21q22.3, tagged by the single nucleotide polymorphism (SNP) rs35418111, being associated with breast cancer risk at a genome-wide significance level; however, the underlying causal functional variants and gene(s) responsible for this association are unknown. We performed functional genomic analyses to identify potential functional variants and target genes that may mediate this association. Functional annotation for SNPs in high linkage disequilibrium (LD, r > 0.8) with rs35418111 in Asians showed evidence of promoter and/or enhancer activities, including rs35418111, rs2078203, rs8134832, rs57385578, and rs8126917. These five variants were assessed for interactions with nuclear proteins by electrophoretic mobility shift assays. Our results showed that the risk alleles for rs2078203 and rs35418111 altered DNA-protein interaction patterns. Cis-expression quantitative loci (cis-eQTL) analysis, using data from the Genotype-Tissue Expression database (GTEx) European-ancestry female normal breast tissue, indicated that the risk allele of rs35418111 was associated with a decreased expression of the gene, a relatively uncharacterized endoribonuclease in humans. We investigated the biological effects of on breast cancer cell lines by transient knock-down of expression in MCF-7, T47D, and MDA-MB-231 cell lines. Knockdown of mRNA in breast cancer cell lines consistently decreased cell proliferation, colony formation, and migration/invasion, regardless of estrogen receptor status. We performed RNA sequencing in MDA-MB-231 cells transfected with siRNA targeting and subsequent gene set enrichment analysis to identify gene networks associated with knockdown. These data indicated was involved in networks associated with inflammation and metabolism. Finally, we showed trends in expression patterns in breast tissues from The Cancer Genome Atlas (TCGA); early-stage breast cancers had elevated expression compared with normal tissue, but significantly decreased expression in late-stage disease. Our study provides evidence of a significant role for the human gene in breast cancer pathogenesis and the association between the rs35418111/21q22.3 locus and breast cancer risk, which may be mediated through functional SNPs, rs35418111 and rs2078203, that regulate expression of .
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http://dx.doi.org/10.3390/cancers13092037DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8122893PMC
April 2021

Proteomics reveals the function reverse of MPSSS-treated prostate cancer-associated fibroblasts to suppress PC-3 cell viability via the FoxO pathway.

Cancer Med 2021 04 11;10(7):2509-2522. Epub 2021 Mar 11.

Key Laboratory of Protein and Peptide Pharmaceuticals & Laboratory of Proteomics, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China.

Prostate cancer-associated fibroblasts (prostate CAFs) are essential components of the tumor microenvironment and can promote tumor progression through their immunosuppressive functions. MPSSS, a novel polysaccharide purified from Lentinus edodes, has been reported to have anti-tumor activity. MPSSS could also inhibit the immunosuppressive function of prostate CAFs, which has been demonstrated through that the secretome of MPSSS-treated prostate CAFs could inhibit the proliferation of T cells. However, how the secretome of MPSSS-treated prostate CAFs influence prostate cancer progression is still unclear. Interestingly, we found that the low molecular weight (3-100kD) secretome of prostate CAFs (lmwCAFS) could promote the growth of PC-3 cells, while that of MPSSS-treated prostate CAFs (MT-lmwCAFS) could inhibit their growth. We carried out comparative secretomic analysis of lmwCAFS and MT-lmwCAFS to identify functional molecules that inhibit the growth of PC-3 cells, and proteomic analysis of lmwCAFS-treated PC-3 cells and MT-lmwCAFS-treated PC-3 cells to investigate the underlying molecular mechanism. These analyses suggest that TGF-β3 from MT-lmwCAFS may inhibit the growth of PC-3 cells. The validated experiments revealed that TGF-β3 from MT-lmwCAFS activated p21 expression in PC-3 cells by regulating the FoxO pathway thereby inducing G0/G1 cell cycle arrest of PC-3 cells. Overall, our data demonstrated that MPSSS reversed the ability of prostate CAFs to suppress the cell viability of PC-3 cells, which might provide a potential therapeutic strategy to prevent prostate cancer progression.
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http://dx.doi.org/10.1002/cam4.3825DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7982613PMC
April 2021

Oleic Acid and Eicosapentaenoic Acid Reverse Palmitic Acid-induced Insulin Resistance in Human HepG2 Cells via the Reactive Oxygen Species / JUN Pathway.

Genomics Proteomics Bioinformatics 2021 Feb 22. Epub 2021 Feb 22.

Key Laboratory of Protein and Peptide Pharmaceuticals & Laboratory of Proteomics, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China. Electronic address:

The monounsaturated fatty acid (MUFA) oleic acid (OA) has previously been shown to reverse saturated fatty acid palmitic acid (PA)-induced hepatic insulin resistance (IR). However, its underlying molecular mechanism is unclear. In addition, previous studies have also shown that the ω-3 polyunsaturated fatty acid (PUFA) eicosapentaenoic acid (EPA) reverses PA-induced muscle IR, but whether EPA plays the same role in hepatic IR and its possible mechanism involved need to be further clarified. Here, we confirmed that EPA reversed PA-induced IR in HepG2 cells and compared the proteomic changes after treatment with different free fatty acids (FFAs). A total of 234 proteins were determined to be differentially expressed after MUFA treatment. Their functions were mainly related to responses to stress and endogenous stimuli, lipid metabolic process, and protein binding. For PUFA treatment, the PA-induced expression changes of 1326 proteins could be reversed by EPA treatment, 415 of which were mitochondrial proteins, with most of the functional proteins involved in oxidative phosphorylation (OXPHOS) and tricarboxylic acid (TCA) cycle. Mechanistic studies revealed that the protein encoded by JUN and reactive oxygen species (ROS) play a role in OA- and EPA-reversed PA-induced IR, respectively. EPA or OA alleviated PA-induced abnormal adenosine triphosphate (ATP) production, ROS generation, and calcium (Ca) content. Importantly, HO-activated production of ROS increased the protein expression of JUN, further resulting in IR in HepG2 cells. Taken together, we demonstrate that ROS/JUN is a common response pathway employed by HepG2 cells toward FFA-regulated IR.
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http://dx.doi.org/10.1016/j.gpb.2019.06.005DOI Listing
February 2021

Noncoding RNAs in subchondral bone osteoclast function and their therapeutic potential for osteoarthritis.

Arthritis Res Ther 2020 11 25;22(1):279. Epub 2020 Nov 25.

Department of Orthopaedics, Shenzhen Intelligent Orthopaedics and Biomedical Innovation Platform, Guangdong Artificial Intelligence Biomedical Innovation Platform, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University Health Science Center, Shenzhen, 518035, China.

Osteoclasts are the only cells that perform bone resorption. Noncoding RNAs (ncRNAs) are crucial epigenetic regulators of osteoclast biological behaviors ranging from osteoclast differentiation to bone resorption. The main ncRNAs, including miRNAs, circRNAs, and lncRNAs, compose an intricate network that influences gene transcription processes related to osteoclast biological activity. Accumulating evidence suggests that abnormal osteoclast activity leads to the disturbance of subchondral bone remodeling, thus initiating osteoarthritis (OA), a prevalent joint disease characterized mainly by cartilage degradation and subchondral bone remodeling imbalance. In this review, we delineate three types of ncRNAs and discuss their related complex molecular signaling pathways associated with osteoclast function during bone resorption. We specifically focused on the involvement of noncoding RNAs in subchondral bone remodeling, which participate in the degradation of the osteochondral unit during OA progression. We also discussed exosomes as ncRNA carriers during the bone remodeling process. A better understanding of the roles of ncRNAs in osteoclast biological behaviors will contribute to the treatment of bone resorption-related skeletal diseases such as OA.
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http://dx.doi.org/10.1186/s13075-020-02374-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7690185PMC
November 2020

Separation and characterization of extracellular vesicles from human plasma by asymmetrical flow field-flow fractionation.

Anal Chim Acta 2020 Aug 11;1127:234-245. Epub 2020 Jul 11.

Laboratory of Protein and Peptide Pharmaceuticals & Proteomics Laboratory, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China; University of Chinese Academy of Sciences, Beijing, 100049, China. Electronic address:

It is a big challenge to isolate extracellular vesicles (EVs) from human plasma because of the contamination from high abundant lipoproteins, such as high density lipoprotein (HDL) and low density lipoprotein particles (LDL). In this study, the parameters of asymmetrical flow field-flow fractionation (AF4) technology and sample preparation, including cross flow gradient, focusing time, ultrafiltration condition, sample amount and injection volume have been optimized and successfully utilized for the separation and characterization of EVs from human plasma. This study demonstrated that the great potential of AF4 in the separation of EVs from HDL and LDL in human plasma with high reproducibility and purity. This study indicated excessive focusing time in the AF4 separation and 100-300 kDa MWCO membrane based ultrafiltration in the pre-preparation will cause loss of EVs. A total of 1038 proteins have been identified in seven replicates of purified EVs from pooled human plasma sample. They are mainly enriched in extracellular exosomes, involved in extracellular matrix structural constituent, and associated with extracellular matrix-receptor interaction pathway. This study also indicated that human plasma contains more EVs than the paired serum at the same volume, and showed age- and gender-independent individual variability of the amount of EVs in human plasma. This study displayed that AF4 technique can serve as a powerful platform for the separation of EVs from human plasma, serum or human body fluids and this technology will promote the studies on EVs, such as proteomics, biomarker discovery and functions.
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http://dx.doi.org/10.1016/j.aca.2020.06.071DOI Listing
August 2020

Integrating metabolomics and network pharmacology to explore the protective effect of gross saponins of Tribulus terrestris L. fruit against ischemic stroke in rat.

J Ethnopharmacol 2020 Dec 5;263:113202. Epub 2020 Aug 5.

Key Laboratory of Medicinal Materials, Jilin Academy of Chinese Medicine Sciences, Changchun, Jilin, 130021, China. Electronic address:

Ethnopharmacological Relevance: Tribulus terrestris L. belongs to the family Zygophyllaceae and has been widely used as a folk medicine for a long history in Asian countries. Gross saponins of Tribulus terrestris L. fruit (GSTTF) has an obvious neuroprotective effect on the treatment of ischemic stroke, but its potential therapeutic mechanisms have not been thoroughly studied.

Aim Of The Study: To investigate the protective effect of GSTTF against ischemic stroke in rat.

Materials And Methods: The combination of metabolomics and network pharmacology analysis was applied to investigate the protective effects of GSTTF on ischemic stroke and its putative mechanism. The related pathway of the biomarkers highlighted from metabolomics analysis was explored, then the possible targets of GSTTF were further revealed by network pharmacology analysis. Molecular docking was conducted to investigate the interaction between the active compound and target protein.

Results: Metabolomics analysis showed that metabolic disturbances were observed in serum for the rats in middle cerebral artery occlusion (MCAO). These MCAO-induced deviations in serum metabolism can be reversely changed by GSTTF via metabolic pathways regulation. Twenty-four proteins with the connectivity degree larger than 15 were selected by the network pharmacology analysis, which are considered as the possible therapeutic targets of the GSTTF against ischemic stroke. The results of molecular docking showed that the active compounds were capable of binding to the representative potential targets HSD11B1 and AR, respectively. And the docking mode of two compounds with the lowest binding energy to their target protein was illustrated by the ribbon binding map.

Conclusion: The present study combines metabolomics and network pharmacology analysis to investigate the mechanism of MCAO-induced ischemic stroke and reveal the efficiency and possible mechanisms of GSTTF for ischemic stroke. Further studies on the bioactive saponin as well as their synergistic action on ischemic stroke will be conducted to better reveal the underlying mechanisms.
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http://dx.doi.org/10.1016/j.jep.2020.113202DOI Listing
December 2020

Endovascular aortic repairs combined with looping-chimney technique for repairing aortic arch lesions and reconstructing left common carotid artery.

J Thorac Dis 2020 May;12(5):2270-2279

Key Laboratory of Cardiovascular Disease of Yunnan Province, Kunming 650051, China.

Background: The aim of this retrospective study was to evaluate the feasibility and efficacy of thoracic endovascular aortic repairs (TEVAR) combined with looping chimney technique (LCT) for repairing aortic arch lesions and reconstructing left common carotid artery.

Methods: Total of 14 patients (mean age 52.86±14.46 years; range, 27-79; 10 men, 4 women) were included in the study from December 2016 to December 2018. Aortic arch pathologies of all patients with insufficient proximal landing zone (PLZ) were repaired by TEVAR under local anesthesia, before TEVAR, the left common carotid artery (LCCA) was protected by the guiding sheath from the retrograde brachial access, after aortic stent graft deployed, chimney graft was implanted to restore LCCA by LCT if necessary. All patients underwent computed tomography angiograph (CTA) 2 weeks, 3 months, 6 months and 1 year after surgery.

Results: Pathology results of 14 patients included: type B aortic dissection (n=8), penetrating aortic ulcers (n=1), retrograde type A aortic dissection (n=1), thoracic aortic aneurysm (TAA) (n=2), and thoracic aortic pseudoaneurysm (n=2). In all patients, aortic arch lesions were repaired by TEVAR; while LCCA were successfully reconstructed by the LCT. In one case, the innominate artery (IA) was simultaneously reconstructed through the same percutaneous right brachial artery (RBA) access. Coiling eliminated type Ia endoleak in 3 patients, and type II endoleak vanished by plugging left subclavian artery (LSA) in 2 patients. In four patients, the chimney stent (CG) of LCCA was partially compressed and then another bare stent was implanted to restore patency rate. The mean follow-up duration was 9.77±6.64 months (range, 0-24) and no combinations were observed in 13 patients; except in one patient who died of cerebral hemorrhage due to abnormal coagulation function.

Conclusions: TEVAR combined with LCT has shown to be suitable surgical approach for aortic arch lesions. Either covered intentionally or inadvertently, the LCCA could be safely and effectively reconstructed via percutaneous RBA access. Short-term follow-up demonstrated satisfactory morbidity and mortality in high-risk patients; however, longer follow-up is required to assess the effectiveness and durability of this innovative endovascular procedure.
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http://dx.doi.org/10.21037/jtd.2020.04.31DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7330391PMC
May 2020

Sequential Precipitation and Delipidation Enables Efficient Enrichment of Low-Molecular Weight Proteins and Peptides from Human Plasma.

J Proteome Res 2020 08 3;19(8):3340-3351. Epub 2020 Jul 3.

Key Laboratory of Protein and Peptide Pharmaceuticals & Laboratory of Proteomics, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.

Low-molecular weight proteins and peptides (LMWPs, <30 kDa) in human plasma serve as potential biomarkers or drug targets and are endowed with desirable traits for biological and clinical studies. However, the identification of LMWPs from plasma is retarded by high-abundance proteins, high-molecular weight proteins, and lipids. Here, we present a sequential precipitation and delipidation (SPD) method for the efficient enrichment of LMWPs based on methyl--butyl ether/methanol/water systems. The enriched LMWP sample was analyzed by single-shot liquid chromatography-tandem mass spectrometry employing both HCD and EThcD without tryptic digestion, and 725 peptides were identified on average. The LMWP sample was also digested and analyzed using a bottom-up proteomics pipeline, and 289 proteins were identified, of which 129 (44.6%) proteins were less than 30 kDa and lipoprotein-associated proteins were significantly enriched. Additionally, 25 neuropeptides and 19 long noncoding RNA-encoded polypeptides were identified. Taken together, the SPD method shows good sensitivity and reproducibility when compared with other enrichment methods and has great potential for clinical biomarker discovery and application.
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http://dx.doi.org/10.1021/acs.jproteome.0c00232DOI Listing
August 2020

Short-term effects of nitrogen deposition on nitrogen spatial and temporal distributions in a Calamagrostis angustifolia wetland of the Sanjiang Plain.

PLoS One 2020 21;15(5):e0232767. Epub 2020 May 21.

Harbin Institute of Technology, Harbin, China.

Nitrogen (N) availability is an important factor regulating the feedback mechanisms of global change. This research uses a small Calamagrostis angustifolia wetland i = on the Sanjiang Plain of Northeast China as the research object and 15N tracer technology to study the effects of different nitrogen deposition levels (0 gN/m2, 4 gN/m2, and 8 gN/m2) through in situ controlled field experiments. Temporal and spatial distribution patterns of nitrogen in plants and soils and their short-term effects on nitrous oxide emissions fluxes were studied. The results showed that 1) the nitrogen content in the stems, leaves and roots of C. angustifolia decreased slowly with the growing season. Nitrogen application significantly increased the absorption of tracer nitrogen in the aboveground and underground plant parts (P<0.01), and the more nitrogen applied, the larger the absorption amount was (P<0.01). The absorbed amount accounted for 52%-86% of the total tracer nitrogen. 2) The tracer nitrogen in the soil did not show a significant change; the more nitrogen that was applied, the more nitrogen that was retained in the soil, and the tracer nitrogen adsorbed by the soil was mainly ammonium nitrogen. 3) The variation in the 15N-labeled nitric oxide emissions flux under different nitrogen treatments was consistent; nitrogen application increased the 15N-labeled nitric oxide emissions flux, but the difference between the low-nitrogen and high-nitrogen treatments was not significant (P>0.05).
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0232767PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7241808PMC
July 2020

Enzyme reaction-guided identification of active components from the flowers of Sophora japonica var. violacea.

Food Funct 2020 May 5;11(5):4356-4362. Epub 2020 May 5.

School of Biological Engineering, Dalian Polytechnic University, 1 Qinggongyuan, Dalian 116034, China.

The flower of S. japonica is a favorite food and used as traditional medicine. In the present study, a facile and effective method based on the changes in the composition before and after the enzyme reaction was established to screen the active compounds from complex natural products. The separation of an active compound from the ethanolic extracts of Sophora japonica var. violacea, which exhibited the α-amylase inhibitory activity is presented as an example. The analysis of HPLC showed that one component was reduced by 25% after the enzyme reaction. The potential active compound was isolated via LH-20 gel permeation chromatography and identified as kaempferol 3-O-α-l-rhamnopyranosyl-(1 → 6)-β-d-galactopyranosyl-7-O-α-l-rhamnopyranoside by H and C NMR. The in vitro test indicated that the compound had the α-amylase inhibitory activity, and the IC was 88.56 ± 0.60 μg mL. The molecular docking study of this compound showed that the compound enfolded in the active sites of α-amylase completely and interacted with the amino acid residues through hydrogen bonds, van der Waals force and hydrophobic interactions.
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http://dx.doi.org/10.1039/d0fo00625dDOI Listing
May 2020

Silencing of lncRNA 6030408B16RIK prevents ultrafiltration failure in peritoneal dialysis via microRNA-326-3p-mediated WISP2 down-regulation.

Biochem J 2020 05;477(10):1907-1921

Department of Nephrology, Linyi People's Hospital, Linyi 2760003, P. R. China.

Continuous exposure to peritoneal dialysis (PD) fluid results in peritoneal fibrosis and ultimately causes ultrafiltration failure. Noncoding RNAs, including long noncoding RNAs (lncRNAs) and microRNAs (miRNAs), have been reported to participate in ultrafiltration failure in PD. Therefore, our study aimed to investigate the mechanism of lncRNA 6030408B16RIK in association with miR-326-3p in ultrafiltration failure in PD. Peritoneal tissues were collected from uremic patients with or without PD. A uremic rat model with PD was first established by 5/6 nephrectomy. The relationship between lncRNA 6030408B16RIK, miR-326-3p and WISP2 was identified using luciferase reporter, RNA pull-down and RIP assays. After ectopic expression and depletion treatments in cells, expression of α-SMA, phosphorylated β-catenin, FSP1, E-cadherin and Vimentin was evaluated by RT-qPCR and Western blot analyses, and Collagen III and CD31 expression by immunohistochemistry. Ultrafiltration volume and glucose transport capacity were assessed by the peritoneal equilibration test. Expression of lncRNA 6030408B16RIK and WISP2 was up-regulated and miR-326-3p expression was poor in peritoneal tissues of uremic PD patients and model rats. LncRNA 6030408B16RIK competitively bound to miR-326-3p and then elevated WISP2 expression. Silencing of lncRNA 6030408B16RIK and WISP2 or overexpression of miR-326-3p was shown to decrease the expression of α-SMA, phosphorylated β-catenin, FSP1, Vimentin, Collagen III and CD31, while reducing glucose transport capacity and increasing E-cadherin expression and ultrafiltration volume in uremic PD rats. In summary, lncRNA 6030408B16RIK silencing exerts an anti-fibrotic effect on uremic PD rats with ultrafiltration failure by inactivating the WISP2-dependent Wnt/β-catenin pathway via miR-326-3p.
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http://dx.doi.org/10.1042/BCJ20190877DOI Listing
May 2020

Proteomic Changes of Klebsiella pneumoniae in Response to Colistin Treatment and Mutation-Mediated Colistin Resistance.

Antimicrob Agents Chemother 2020 05 21;64(6). Epub 2020 May 21.

Key Laboratory of Protein and Peptide Pharmaceuticals and Laboratory of Proteomics, Institute of Biophysics, Chinese Academy of Sciences, Beijing, China

Polymyxins are increasingly used as the critical last-resort therapeutic options for multidrug-resistant Gram-negative bacteria. Unfortunately, polymyxin resistance has increased gradually over the past few years. Although studies on polymyxin mechanisms are expanding, systemwide analyses of the underlying mechanism for polymyxin resistance and stress response are still lacking. To understand how adapts to colistin (polymyxin E) pressure, we carried out proteomic analysis of a strain cultured with different concentrations of colistin. Our results showed that the proteomic responses to colistin treatment in involve several pathways, including (i) gluconeogenesis and the tricarboxylic acid (TCA) cycle, (ii) arginine biosynthesis, (iii) porphyrin and chlorophyll metabolism, and (iv) enterobactin biosynthesis. Interestingly, decreased abundances of class A β-lactamases, including TEM, SHV-11, and SHV-4, were observed in cells treated with colistin. Moreover, we present comprehensive proteome atlases of paired polymyxin-susceptible and -resistant strains. The polymyxin-resistant strain Ci, a mutant of ATCC BAA 2146, showed a missense mutation in This mutant, which displayed lipid A modification with 4-amino-4-deoxy-l-arabinose (l-Ara4N) and palmitoylation, showed striking increases in the expression of CrrAB, PmrAB, PhoPQ, ArnBCADT, and PagP. We hypothesize that mutations induce elevated expression of the operon and via PmrAB and PhoPQ. Moreover, the multidrug efflux pump KexD, which was induced by mutation, also contributed to colistin resistance. Overall, our results demonstrated proteomic responses to colistin treatment and the mechanism of CrrB-mediated colistin resistance, which may offer valuable information on the management of polymyxin resistance.
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http://dx.doi.org/10.1128/AAC.02200-19DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7269499PMC
May 2020

Large-scale Identification of N-linked Intact Glycopeptides in Human Serum using HILIC Enrichment and Spectral Library Search.

Mol Cell Proteomics 2020 04 26;19(4):672-689. Epub 2020 Feb 26.

Laboratory of Protein and Peptide Pharmaceuticals & Proteomics Laboratory, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China. Electronic address:

Large-scale identification of -linked intact glycopeptides by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) in human serum is challenging because of the wide dynamic range of serum protein abundances, the lack of a complete serum N-glycan database and the existence of proteoforms. In this regard, a spectral library search method was presented for the identification of -linked intact glycopeptides from -linked glycoproteins in human serum with target-decoy and motif-specific false discovery rate (FDR) control. Serum proteins were firstly separated into low-abundance and high-abundance proteins by acetonitrile (ACN) precipitation. After digestion, the -linked intact glycopeptides were enriched by hydrophilic interaction liquid chromatography (HILIC) and a portion of the enriched -linked intact glycopeptides were processed by Peptide-N-Glycosidase F (PNGase F) to generate -linked deglycopeptides. Both -linked intact glycopeptides and deglycopeptides were analyzed by LC-MS/MS. From -linked deglycopeptides data sets, 764 -linked glycoproteins, 1699 -linked glycosites and 3328 unique -linked deglycopeptides were identified. Four types of -linked glycosylation motifs (NXS/T/C/V, X≠P) were used to recognize the -linked deglycopeptides. The spectra of these -linked deglycopeptides were utilized for -linked deglycopeptides library construction and identification of -linked intact glycopeptides. A database containing 739 N-glycan masses was constructed and utilized during spectral library search for the identification of -linked intact glycopeptides. In total, 526 -linked glycoproteins, 1036 -linked glycosites, 22,677 -linked intact glycopeptides and 738 N-glycan masses were identified under 1% FDR, representing the most in-depth serum N-glycoproteome identified by LC-MS/MS at -linked intact glycopeptide level.
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http://dx.doi.org/10.1074/mcp.RA119.001791DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7124471PMC
April 2020

Highly efficient WO-FeO catalysts synthesized using a novel solvent-free method for NH-SCR.

J Hazard Mater 2020 Apr 3;388:121812. Epub 2019 Dec 3.

Faculty of Environmental Science and Engineering, Kunming University of Science and Technology, Kunming, 650500, PR China. Electronic address:

WO-FeO catalysts with various WO contents were synthesized through a facile solvent-free method, satisfying the selective catalytic reduction of NO (NH-SCR). Strikingly, the optimum 30 %WO-FeO catalyst with the largest surface area exhibited the most outstanding catalytic activity, achieving the nearly 100 % NO removal efficiency in a wide temperature window between 225-500 °C, which was better than that of Fe-W series catalysts reported in other studies. In addition, Raman and XPS results proved that the introduction of WO altered the electronic environment of FeO, inducing the formation of FeO (Fe) and surface adsorbed oxygen. In situ DRIFTS demonstrated that the interaction between WO and FeO not only promoted the adsorption capacity of NH on the catalyst, but also contributed to the formation of adsorbed NO species. NO reduction reaction on WO-FeO catalyst proceeded via the Eley-Rideal and Langmuir-Hinshelwood mechanism synchronously. All of these factors, jointly, accounted for the superior catalytic activity and N selectivity of WO-FeO catalysts.
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http://dx.doi.org/10.1016/j.jhazmat.2019.121812DOI Listing
April 2020

Riolan arch pseudoaneurysm hemorrhage after endovascular covered stent-graft treatment of an abdominal aortic aneurysm: A case report.

Medicine (Baltimore) 2019 Nov;98(48):e17789

Department of Vascular Intervention.

Introduction: Riolan arch thickening is usually caused by the occlusion of the superior mesenteric artery (SMA), inferior mesenteric artery, or abdominal aortic artery, by colon cancer, or by ulcerative colitis in the active phase.

Patient Concerns: A 61-years-old female was admitted due to left lower abdominal pain, nausea, and vomiting for more than 4 days. She had received an endovascular covered stent-graft exclusion due to abdominal aortic aneurysm 18 months earlier. Computed tomographic angiography (CTA) showed a local rupture of 1 of the branch artery of the SMA, and a pseudoaneurysm was formed around it. It was feared that performing Riolan atrial arch pseudoaneurysm embolization may cause ischemia of the inferior mesenteric artery (IMA) and could lead to avascular necrosis of the descending colon and sigmoid colon, intestinal perforation, and peritonitis.

Diagnosis: Riolan arch collateral circulation associated with pseudoaneurysm hemorrhage after endovascular covered stent-graft treatment of an abdominal aortic aneurysm.

Interventions: Riolan arterial arch pseudoaneurysm embolization was performed near the distal end.

Outcomes: The symptoms, signs, and biochemistry returned to normal.

Conclusion: Riolan arch collateral circulation can be caused by pseudoaneurysm hemorrhage after endovascular covered stent-graft treatment of an abdominal aortic aneurysm.
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http://dx.doi.org/10.1097/MD.0000000000017789DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6890315PMC
November 2019

Investigating the Protective Effect of Gross Saponins of Fruit against Ischemic Stroke in Rat Using Metabolomics and Network Pharmacology.

Metabolites 2019 Oct 21;9(10). Epub 2019 Oct 21.

Key Laboratory of Medicinal Materials, Jilin Academy of Chinese Medicine Sciences, Changchun 130021, China.

Stroke is one of the leading causes of death and long-term disability worldwide. Gross saponins of fruit (GSTTF) has been used for neuroprotective therapy on convalescents of ischemic stroke. But the related therapeutic mechanisms have not yet been well investigated. This study aimed to investigate the protective effects of GSTTF on ischemic stroke using metabolomics coupled with network pharmacology analysis. The rat urine sample was collected and profiled by an LC-MS-based metabolomics approach. The pathway analysis was performed based on the highlighted biomarkers, then the network pharmacology approach was applied to screen the potential therapeutic targets of GSTTF. Metabolomics analysis showed that a series of metabolic perturbations occurred in the middle cerebral artery occlusion (MCAO) group compared with the sham group. Gross saponins of fruit can change the MCAO-induced urine metabolic deviations in a reverse manner via regulating multiple metabolic pathways. Two proteins, inducible nitric oxide synthase (NOS2) and glycogen synthase kinase-3 beta (GSK3B), were highlighted by the network pharmacology analysis, which may be the potential therapeutic targets for the GSTTF against ischemic stroke. This study provides an overview of the mechanism of MCAO-induced ischemic stroke and investigates the efficacy of GSTTF in the treatment of ischemic stroke. Further study is needed to reveal its underlying mechanisms more clearly.
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http://dx.doi.org/10.3390/metabo9100240DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6835270PMC
October 2019

Ginsenoside compound K ameliorates Alzheimer's disease in HT22 cells by adjusting energy metabolism.

Mol Biol Rep 2019 Oct 30;46(5):5323-5332. Epub 2019 Jul 30.

Laboratory of Molecular Pharmacology, Jilin Provincial Key Laboratory of BioMacromolecules of Chinese Medicine, Jilin Ginseng Academy, Changchun University of Chinese Medicine, Changchun, 130117, Jilin, China.

Energy metabolism disorders have been shown to exert detrimental effects on the pathology of Alzheimer's disease (AD). The ginsenoside compound K (CK), a major intestinal metabolite underlying the pharmacological actions of orally administered ginseng, has an ameliorating effect against AD, but the relevant molecular mechanism remains unclear. We hypothesized that the improvement of AD by CK is mediated by the energy metabolism signaling pathway induced by amyloid β peptide (Aβ) and tested this hypothesis in HT22 cells. HT22 cells were incubated with CK and exposed to Aβ. Cell viability was analyzed using the MTT assay. Cell growth curves were derived from real-time cell analysis. Apoptosis was determined by flow cytometry, Aβ localization and expression by immunofluorescence, and ATP content by a specific assay kit. The expression of proteins related to the energy metabolism signaling pathway was analyzed using Western blotting. CK treatment improved cell viability, cell growth, and apoptosis induced by Aβ, and the cellular localization and expression of Aβ. Moreover, CK increased ATP content by promoting the activity of glucose transporters (GLUTs). Therefore, the neuroprotective effect of CK against Aβ injury was mainly realized through the activation of the energy metabolism signaling pathway. CK treatment inhibits neuronal damage caused by Aβ through the activation of the energy metabolism signaling pathway, revealing that CK might be one of the key bioactive ingredients of ginseng in the treatment of Alzheimer's disease and may serve as a preventive or therapeutic agent for Alzheimer's disease.
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http://dx.doi.org/10.1007/s11033-019-04988-0DOI Listing
October 2019

Comprehensive characterization and proteoform analysis of the hydrophobic surfactant proteins B and C in calf pulmonary surfactant.

J Pharm Biomed Anal 2019 Sep 21;174:625-632. Epub 2019 Jun 21.

Key Laboratory of Protein and Peptide Pharmaceuticals & Laboratory of Proteomics, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China. Electronic address:

Calf pulmonary surfactant (CPS), which contains about 98% lipids and 2% hydrophobic surfactant proteins B (SP-B) and C (SP-C), has been used as a surfactant preparation for the clinical replacement therapy of respiratory distress syndrome (RDS). Characterization of SP-B and SP-C in CPS is informative for quality control and the evaluation of their biological activities. However, analysis of SP-B and SP-C is impeded by the high content of lipids in CPS. Here, we describe an integrated method by combining size exclusion chromatography (SEC)-based delipidation, SDS-PAGE separation, in-gel digestion and mass spectrometric analysis for comprehensive characterization and proteoform analysis of the extremely hydrophobic SP-B and SP-C in CPS. This study has shown that 30 proteoforms of SP-C with different truncations and modifications were identified and SP-B was found to be existed as a dimer form in the CPS.
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http://dx.doi.org/10.1016/j.jpba.2019.06.027DOI Listing
September 2019

Atlastin-mediated membrane tethering is critical for cargo mobility and exit from the endoplasmic reticulum.

Proc Natl Acad Sci U S A 2019 07 25;116(28):14029-14038. Epub 2019 Jun 25.

National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 100101 Beijing, China;

Endoplasmic reticulum (ER) membrane junctions are formed by the dynamin-like GTPase atlastin (ATL). Deletion of ATL results in long unbranched ER tubules in cells, and mutation of human ATL1 is linked to hereditary spastic paraplegia. Here, we demonstrate that COPII formation is drastically decreased in the periphery of ATL-deleted cells. ER export of cargo proteins becomes defective; ER exit site initiation is not affected, but many of the sites fail to recruit COPII subunits. The efficiency of cargo packaging into COPII vesicles is significantly reduced in cells lacking ATLs, or when the ER is transiently fragmented. Cargo is less mobile in the ER in the absence of ATL, but the cargo mobility and COPII formation can be restored by ATL R77A, which is capable of tethering, but not fusing, ER tubules. These findings suggest that the generation of ER junctions by ATL plays a critical role in maintaining the necessary mobility of ER contents to allow efficient packaging of cargo proteins into COPII vesicles.
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http://dx.doi.org/10.1073/pnas.1908409116DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6628656PMC
July 2019

Identification of Novel Susceptibility Loci and Genes for Prostate Cancer Risk: A Transcriptome-Wide Association Study in Over 140,000 European Descendants.

Cancer Res 2019 07 17;79(13):3192-3204. Epub 2019 May 17.

Division of Epidemiology, Department of Medicine, Vanderbilt Epidemiology Center, Vanderbilt-Ingram Cancer Center, Vanderbilt University Medical Center, Nashville, Tennessee.

Genome-wide association study-identified prostate cancer risk variants explain only a relatively small fraction of its familial relative risk, and the genes responsible for many of these identified associations remain unknown. To discover novel prostate cancer genetic loci and possible causal genes at previously identified risk loci, we performed a transcriptome-wide association study in 79,194 cases and 61,112 controls of European ancestry. Using data from the Genotype-Tissue Expression Project, we established genetic models to predict gene expression across the transcriptome for both prostate models and cross-tissue models and evaluated model performance using two independent datasets. We identified significant associations for 137 genes at < 2.61 × 10, a Bonferroni-corrected threshold, including nine genes that remained significant at < 2.61 × 10 after adjusting for all known prostate cancer risk variants in nearby regions. Of the 128 remaining associated genes, 94 have not yet been reported as potential target genes at known loci. We silenced 14 genes and many showed a consistent effect on viability and colony-forming efficiency in three cell lines. Our study provides substantial new information to advance our understanding of prostate cancer genetics and biology. SIGNIFICANCE: This study identifies novel prostate cancer genetic loci and possible causal genes, advancing our understanding of the molecular mechanisms that drive prostate cancer.
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http://dx.doi.org/10.1158/0008-5472.CAN-18-3536DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6606384PMC
July 2019

Occupational manganese exposure, reproductive hormones, and semen quality in male workers: A cross-sectional study.

Toxicol Ind Health 2019 Jan 22;35(1):53-62. Epub 2018 Nov 22.

1 Department of Environmental Health, School of Public Health, China Medical University, Shenyang, China.

It has been found that exposure to manganese (Mn) could induce reproductive dysfunction, but its occupational risk in male workers is unclear. This study aims to assess the association of occupational Mn exposure with reproductive hormones and semen quality in a cross-sectional study. Urinary Mn, semen quality, and reproductive hormones were explored in 84 male workers occupationally exposed to Mn and 92 referents. Multiple linear regression analyses were used to assess the relationship. Urinary Mn levels in Mn-exposed workers ranged from 0.56 to 34.25 µg/L, and the average level was 15.92 ± 8.49 µg/L. Compared with the control group, gonadotropin-releasing hormone (GnRH) levels and luteinizing hormone (LH) levels increased significantly and the levels of testosterone (TSTO) decreased significantly in the Mn-exposed group. There was a significant positive linear association between urinary Mn and GnRH and LH, while the linear association between urinary Mn and TSTO was negative. Sperm progressive motility and total motility decreased significantly in the Mn-exposed group. There was a significantly negative linear association between urinary Mn and sperm progressive motility and total motility. In conclusion, occupational Mn exposure was inversely associated with reproductive health of male workers, resulting in the abnormality of hormones secretion and decrease of sperm motility.
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http://dx.doi.org/10.1177/0748233718810109DOI Listing
January 2019

SIRT5 deacylates metabolism-related proteins and attenuates hepatic steatosis in ob/ob mice.

EBioMedicine 2018 Oct 29;36:347-357. Epub 2018 Sep 29.

National Laboratory of Biomacromolecules, Institute of Biophysics, Cthe Strategic Priority Research Programses, Beijing 100101, China. Electronic address:

Background: Sirtuin 5 (SIRT5) is a NAD-dependent lysine deacylase. The SIRT5 deficiency mouse model shows that it is dispensable for metabolic homeostasis under normal conditions. However, the biological role of SIRT5 and acylation in pathological states such as obesity and type 2 diabetes (T2D) remains elusive.

Methods: The hepatic SIRT5-overexpressing ob/ob mouse model (ob/ob-SIRT5 OE) was established by CRISPR/Cas9 gene editing tool Protein malonylation and succinylation lysine sites were identified by immunoprecipitation coupled lipid chromatography - tandem mass spectrometry (LC-MS/MS) methods.

Findings: The ob/ob-SIRT5 OE mice showed decreased malonylation and succinylation, improved cellular glycolysis, suppressed gluconeogenesis, enhanced fatty acid oxidation, and attenuated hepatic steatosis. A total of 955 malonylation sites on 434 proteins and 1377 succinylation sites on 429 proteins were identified and quantitated. Bioinformatics analysis revealed that malonylation was the major SIRT5 target in the glycolysis/gluconeogenesis pathway, whereas succinylation was the preferred SIRT5 target in the oxidative phosphorylation pathway.

Interpretation: Hepatic overexpression of SIRT5 ameliorated the metabolic abnormalities of ob/ob mice, probably through demalonylating and desuccinylating proteins in the main metabolic pathways. SIRT5 and related acylation might be potential targets for metabolic disorders. FUND: National Key R&D Program of China, the National Natural Science Foundation of China, the Strategic Priority Research Programs (Category A) of the Chinese Academy of Sciences, the Interdisciplinary Medicine Seed Fund of Peking University and the National Laboratory of Biomacromolecules.
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http://dx.doi.org/10.1016/j.ebiom.2018.09.037DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6197389PMC
October 2018

Author Correction: In vitro inhibition of hepatic stellate cell activation by the autophagy-related lipid droplet protein ATG2A.

Sci Rep 2018 Sep 26;8(1):14569. Epub 2018 Sep 26.

First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.
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http://dx.doi.org/10.1038/s41598-018-32560-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6158263PMC
September 2018

N-Linked Glycopeptide Identification Based on Open Mass Spectral Library Search.

Biomed Res Int 2018 14;2018:1564136. Epub 2018 Aug 14.

National Center for Mathematics and Interdisciplinary Sciences, Key Laboratory of Random Complex Structures and Data Science, Academy of Mathematics and Systems Science, Chinese Academy of Sciences, Beijing 100101, China.

Confident characterization of intact glycopeptides is a challenging task in mass spectrometry-based glycoproteomics due to microheterogeneity of glycosylation, complexity of glycans, and insufficient fragmentation of peptide bones. Open mass spectral library search is a promising computational approach to peptide identification, but its potential in the identification of glycopeptides has not been fully explored. Here we present pMatchGlyco, a new spectral library search tool for intact N-linked glycopeptide identification using high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS) data. In pMatchGlyco, (1) MS/MS spectra of deglycopeptides are used to create spectral library, (2) MS/MS spectra of glycopeptides are matched to the spectra in library in an open (precursor tolerant) manner and the glycans are inferred, and (3) a false discovery rate is estimated for top-scored matches above a threshold. The efficiency and reliability of pMatchGlyco were demonstrated on a data set of mixture sample of six standard glycoproteins and a complex glycoprotein data set generated from human cancer cell line OVCAR3.
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http://dx.doi.org/10.1155/2018/1564136DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6112209PMC
December 2018

Arbuscular Mycorrhizal Fungi () Improves Growth, Photosynthesis and Protects Photosystem II in Leaves of L. in Cadmium Contaminated Soil.

Front Plant Sci 2018 13;9:1156. Epub 2018 Aug 13.

College of Life Science, Northeast Forestry University, Harbin, China.

In this study, the effects of inoculating arbuscular mycorrhizal fungi () on the growth, chlorophyll content, photosynthetic gas exchange parameters, and chlorophyll fluorescence characteristics of L. in cadmium (Cd) contaminated soil were investigated. The results showed that the root vigor of declined, while the chlorophyll content significantly decreased with the increase of Cd content, especially the chlorophyll a content in leaves. The photosynthetic carbon assimilation capacity and PSII activity of leaves were also significantly inhibited by Cd stress, especially the electron transfer at the receptor side of PSII, which was more sensitive to Cd stress. The infection level of on roots was relatively high and inoculation with increased the mycorrhizal infection rate of roots up to 50-70%. Due to the impact of the mycorrhizal infection, the Cd content in roots was significantly increased compared to non-inoculated treatment; however, the Cd content in the aboveground part of was not significantly different compared to the non-inoculated treatment. After inoculation with , the root vigor of increased to some extent, alleviating the chlorophyll degradation in leaves under Cd contaminated soil. Infection with can improve the stoma limitation of leaves in Cd contaminated soil and increase the non-stomatal factors including the tolerance of its photosynthetic apparatus to Cd, to improve photosynthetic capacity. infection can improve the photosynthetic electron transport capacity of PSII in leaves under Cd stress and promotes the activity of the oxygen-evolving complex to different degrees at the donor side of PSII and the electron transport capacity from Q to Q on the receptor side of PSII. Thus, this guarantees that leaves inoculated with in Cd contaminated soil have relatively higher PSII activity. Therefore, inoculation with can improve the capacity of Cd tolerance of with regard to various aspects, such as morphological characteristics and photosynthetic functions, and reduce the toxicity of Cd on .
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http://dx.doi.org/10.3389/fpls.2018.01156DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6099091PMC
August 2018

Temporal Proteomic Analysis of Pancreatic β-Cells in Response to Lipotoxicity and Glucolipotoxicity.

Mol Cell Proteomics 2018 11 6;17(11):2119-2131. Epub 2018 Aug 6.

From the ‡National Laboratory of Biomacramolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China;

Chronic hyperlipidemia causes the dysfunction of pancreatic β-cells, such as apoptosis and impaired insulin secretion, which are aggravated in the presence of hyperglycemia. The underlying mechanisms, such as endoplasmic reticulum (ER) stress, oxidative stress and metabolic disorders, have been reported before; however, the time sequence of these molecular events is not fully understood. Here, using isobaric labeling-based mass spectrometry, we investigated the dynamic proteomes of INS-1 cells exposed to high palmitate in the absence and presence of high glucose. Using bioinformatics analysis of differentially expressed proteins, including the time-course expression pattern, protein-protein interaction, gene set enrichment and KEGG pathway analysis, we analyzed the dynamic features of previously reported and newly identified lipotoxicity- and glucolipotoxicity-related molecular events in more detail. Our temporal data highlight cholesterol metabolism occurring at 4 h, earlier than fatty acid metabolism that started at 8 h and likely acting as an early toxic event highly associated with ER stress induced by palmitate. Interestingly, we found that the proliferation of INS-1 cells was significantly increased at 48 h by combined treatment of palmitate and glucose. Moreover, benefit from the time-course quantitative data, we identified and validated two new molecular targets: Setd8 for cell replication and Rhob for apoptosis, demonstrating that our temporal dataset serves as a valuable resource to identify potential candidates for mechanistic studies of lipotoxicity and glucolipotoxicity in pancreatic β-cells.
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http://dx.doi.org/10.1074/mcp.RA118.000698DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6210228PMC
November 2018

In vitro inhibition of hepatic stellate cell activation by the autophagy-related lipid droplet protein ATG2A.

Sci Rep 2018 06 18;8(1):9232. Epub 2018 Jun 18.

First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China.

Clinical studies have found that moderate intake of retinol or oleic acid can enlarge the lipid droplets of hepatic stellate cells and suppress their activation. However, the link between lipid droplets and cell activation is unknown. This study compared the dynamics of lipid droplet-associated protein expression between activated and reverted stellate cells. Reversion of the activated human stellate cell line LX-2 and inhibition of primary mouse stellate cell activation were induced by retinol or oleic acid, which resulted in larger lipid droplets and the downregulation of cell activation markers. Quantitative proteomics and immunoblotting were performed to compare lipid-droplet protein profiles between activated and reverted LX-2 cells. Compared to expression in activated cells, 50 lipid-droplet proteins were upregulated, whereas 28 were downregulated upon reversion. ATG2A was significantly enriched in lipid droplets of retinol/oleic acid-treated LX-2 cells and quiescent primary stellate cells. Reduced expression of α-SMA, increased expression of perilipin-3, enlarged lipid droplets, and suppression of autophagic flux were observed in ATG2A-deficient LX2 cells. Lipid-droplet protein profile changes during the reversion of activated stellate cells might provide new insights into the molecular mechanisms linking lipid droplets to liver fibrosis. ATG2A could represent a potential new drug target for hepatic fibrosis.
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http://dx.doi.org/10.1038/s41598-018-27686-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6006255PMC
June 2018