Publications by authors named "Yanliang Wang"

40 Publications

Identification of connexin43 in diabetic retinopathy and its downregulation by O-GlcNAcylation to inhibit the activation of glial cells.

Biochim Biophys Acta Gen Subj 2021 Jul 3;1865(10):129955. Epub 2021 Jul 3.

Department of Ophthalmology, Shanghai Tenth People's Hospital affiliated with Tongji University, 301 Middle Yan Chang Road, Shanghai 200072, PR China. Electronic address:

Background: Despite advances in the treatments of diabetic complications, proliferative diabetic retinopathy (PDR) still remains a major cause leading to visual loss, mainly because of the lack of pathological mechanisms and complicated protein expressions in vivo. Current study aimed to investigate the patterns of connexin43 (Cx43) changes and the possible interactions with O-GlcNAcylation in DR.

Methods: Clinical samples of vitreous and fibrovascular membranes were acquired from PDR patients during pars plana vitrectomy. Brown Norway rats were used to build diabetic animal models; to investigate the effects of O-GlcNAcylation on Cx43 expressions, total retinal O-GlcNAcylation was changed by intravitreal injections. Levels of protein expressions were examined by immunofluorescence staining and western blot.

Results: Our results revealed increased Cx43 expressions in a vessel-shape pattern followed by the distribution of glial fibrillary acidic protein (GFAP) in diabetic fibrovascular membranes. Similarly, Cx43 and GFAP expressions were elevated in PDR vitreous and diabetic animal retinas. Retinal O-GlcNAcylation was effectively regulated by intravitreal injections, and the increase of Cx43 and GFAP was significantly suppressed by O-GlcNAcylation inhibition under hyperglycemia conditions.

Conclusions: We systemically proved the changes of Cx43 with different retinal cells, and reported the effective methods to regulate retinal O-GlcNAcylation by intravitreal injections, and clearly illustrated the downregulated effects of O-GlcNAcylation inhibition on Cx43 and GFAP expressions.

General Significance: Targeting connexin43 in glial cells reveals a novel mechanism to understand the formation of diabetic fibrovascular membranes and offers a potential therapeutic strategy to interfere the development of PDR.
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http://dx.doi.org/10.1016/j.bbagen.2021.129955DOI Listing
July 2021

The clear cell sarcoma functional genomic landscape.

J Clin Invest 2021 Jun 22. Epub 2021 Jun 22.

Department of Orthopedics and Oncological Sciences, University of Utah, Salt Lake City, United States of America.

Clear Cell Sarcoma (CCS) is a deadly malignancy affecting adolescents and young adults. It is characterized by reciprocal translocations resulting in the expression of the chimeric EWSR1-ATF1 or EWSR1-CREB1 fusion proteins, driving sarcomagenesis. Besides these characteristics, CCS has remained genomically uncharacterized. Copy number analysis of human CCSs showed frequent amplifications of the MITF locus and chromosomes 7 and 8. Few alterations were shared with Ewing sarcoma or desmoplastic small round cell tumors, other EWSR1-rearranged tumors. Exome sequencing in mouse tumors generated by expressing EWSR1-ATF1 from the Rosa26 locus demonstrated no other repeated pathogenic variants. Additionally, we generated a new CCS mouse by Cre-loxP-induced chromosomal translocation between Ewsr1 and Atf1, resulting in copy number loss of chromosome 6 and chromosome 15 instability, including amplification of a portion syntenic with human chromosome 8, surrounding Myc. Additional experiments in the Rosa26 conditional model demonstrated that Mitf or Myc can contribute to sarcomagenesis. Copy number observations in human tumors and genetic experiments in mice render, for the first time, a functional landscape of the CCS genome. These data advance efforts to understand the biology of CCS with innovative models, in which we can eventually validate preclinical therapies, necessary to move toward longer and better survival of the young victims of this disease.
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http://dx.doi.org/10.1172/JCI146301DOI Listing
June 2021

Reduction in pericyte coverage leads to blood-brain barrier dysfunction via endothelial transcytosis following chronic cerebral hypoperfusion.

Fluids Barriers CNS 2021 May 5;18(1):21. Epub 2021 May 5.

Department of Neurology, Henan Provincial People's Hospital, Zhengzhou University People's Hospital, Henan University People's Hospital, Zhengzhou, 450003, Henan, China.

Background: Chronic cerebral hypoperfusion (CCH) is the leading cause of cerebral small vessel disease (CSVD). CCH is strongly associated with blood-brain barrier (BBB) dysfunction and white matter lesions (WMLs) in CSVD. However, the effects of CCH on BBB integrity and components and the cellular and molecular mechanisms underlying the effects of BBB dysfunction remain elusive. Whether maintaining BBB integrity can reverse CCH-induced brain damage has also not been explored.

Methods: In this study, we established a rat model of CSVD via permanent bilateral common carotid artery occlusion (2VO) to mimic the chronic hypoperfusive state of CSVD. The progression of BBB dysfunction and components of the BBB were assessed using immunostaining, Western blotting, transmission electron microscopy (TEM) and RNA sequencing. We also observed the protective role of imatinib, a tyrosine kinase inhibitor, on BBB integrity and neuroprotective function following CCH. The data were analyzed using one-way or two-way ANOVA.

Results: We noted transient yet severe breakdown of the BBB in the corpus callosum (CC) following CCH. The BBB was severely impaired as early as 1 day postoperation and most severely impaired 3 days postoperation. BBB breakdown preceded neuroinflammatory responses and the formation of WMLs. Moreover, pericyte loss was associated with BBB impairment, and the accumulation of serum protein was mediated by increased endothelial transcytosis in the CC. RNA sequencing also revealed increased transcytosis genes expression. BBB dysfunction led to brain damage through regulation of TGF-β/Smad2 signaling. Furthermore, imatinib treatment ameliorated serum protein leakage, oligodendrocyte progenitor cell (OPC) activation, endothelial transcytosis, microglial activation, and aberrant TGF-β/Smad2 signaling activation.

Conclusions: Our results indicate that reduced pericyte coverage leads to increased BBB permeability via endothelial transcytosis. Imatinib executes a protective role on the BBB integrity via inhibition of endothelial transcytosis. Maintenance of BBB integrity ameliorates brain damage through regulation of TGF-β/Smad2 signaling following CCH; therefore, reversal of BBB dysfunction may be a promising strategy for CSVD treatment.
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http://dx.doi.org/10.1186/s12987-021-00255-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8101037PMC
May 2021

Expression and clinical significance of pyruvate kinase M2 in breast cancer: A protocol for meta-analysis and bioinformatics validation analysis.

Medicine (Baltimore) 2021 May;100(18):e25545

Department of General Surgery, Hainan Western Central Hospital, Danzhou 571799, Hainan Province, China.

Background: Breast cancer is a common malignant tumor in women. In recent years, its incidence is increasing year by year, and its morbidity and mortality rank the first place among female malignant tumors. Some key enzymes and intermediates in glycolysis are closely related to tumor development. Pyruvate kinase M2 (PKM2) is an important rate-limiting enzyme in glycolysis pathway. Meanwhile, it is highly expressed in proliferative cells, especially in tumor cells, and plays an important role in the formation of Warburg effect and tumorigenesis. Previous studies have explored the effects of PKM2 expression on the prognosis and clinical significance of breast cancer patients, while the results are contradictory and uncertain. This study uses controversial data for meta-analysis to accurately evaluate the problem. We collected relevant Oncomine and The Cancer Genome Atlas (TCGA) data to further verify the results. Through bioinformatics analysis, the mechanism and related pathways of PKM2 in breast cancer are explored.

Methods: We searched Wanfang, Chinese Biomedical Literature Database, Chinese National Knowledge Infrastructure, the Chongqing VIP Chinese Science and Technology Periodical Database, PubMed, Embase, and Web of Science databases from inception to March 2021. The language restrictions are Chinese and English. The published literatures on PKM2 expression and prognosis or clinicopathological characteristics of breast cancer patients were statistically analyzed. Combined hazard ratios (HRs), odds ratios (ORs), and 95% confidence intervals (95% CIs) were used to evaluate the effects of PKM2 on the prognosis and clinicopathological features of breast cancer. Stata 14.0 software was applied for meta-analysis. Oncomine and TCGA databases were used to meta-analyze the differences of PKM2 mRNA expression between breast cancer and normal breast tissues. The expression of PKM2 protein was verified by Human Protein Atlas (HPA) database. The relationship between the gene and the survival of breast cancer patients was analyzed by Gene Expression Profiling Interactive Analysis (GEPIA). The relationship between PKM2 gene and clinicopathological characteristics was analyzed by using LinkedOmics, and the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment pathway analysis was performed by using Metascape. Protein-protein interaction (PPI) network was constructed by String website.

Results: The results of this meta-analysis will be submitted to a peer-reviewed journal for publication.

Conclusion: This study provides high-quality medical evidence for the correlation between the expression of PKM2 and the prognosis and clinicopathological features of breast cancer. Through bioinformatics analysis, this study further deepens the understanding of the mechanism and related pathways of PKM2 in breast cancer.

Ethics And Dissemination: The private information from individuals will not be published. This systematic review also should not damage participants' rights. Ethical approval is not available. The results may be published in a peer-reviewed journal or disseminated in relevant conferences.

Osf Registration Number: DOI 10.17605/OSF.IO/W52HB.
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http://dx.doi.org/10.1097/MD.0000000000025545DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8104244PMC
May 2021

Global Characterization of Immune Infiltration in Clear Cell Renal Cell Carcinoma.

Onco Targets Ther 2021 22;14:2085-2100. Epub 2021 Mar 22.

Henan Provincial Key Laboratory of Kidney Disease and Immunology, Henan Provincial People's Hospital, Zhengzhou, 450052, Henan, People's Republic of China.

Background: Immunotherapy has revolutionized the treatment of clear cell renal cell carcinoma (ccRCC). However, the therapy is constrained by drug resistance. Therefore, further characterization of immune infiltration in ccRCC is needed to improve its efficacy.

Methods: Here, we adopted the CIBERSORT method to analyze the level of 22 immune cells, and analyzed the correlation of immune cells and clinical parameters in ccRCC in The Cancer Genome Atlas. We used consensus clustering to cluster ccRCC and identified differently expressed genes (DEGs) between hot and cold tumors using the "Limma" package, and then performed enrichment analysis of DEGs. Finally, we constructed and validated a Cox regression model using the "survival", "glmnet", and "survivalROC" packages, implemented in R.

Results: Regulatory T cells upregulated in tumor tissue increased during tumor progression, and correlated with poor overall survival in ccRCC. Consensus clustering identified four clusters of ccRCC. To elucidate the underlying mechanisms of immune cell infiltration, we subdivided these four clusters into two major types, immune hot and cold, and identified DEGs between them. The results revealed different transcription profiles in the two tumor types, with hot tumors being enriched in immune-related signaling, whereas cold tumors were enriched in extracellular matrix remodeling and the phosphatidylinositol 3-kinase-AKT (PI3K/AKT) pathway. We further identified hub genes and prognostic-related genes from the DEGs, and constructed a Cox regression model for predicting the overall survival of patients with ccRCC. The areas under the receiver operating characteristics curve for the risk model for the training, testing, and external Zhengzhou validation cohorts were 0.834, 0.733, and 0.812, respectively. Notably, gene sets in the prediction model could also predict the overall survival of patients receiving immunotherapy.

Conclusion: These findings provide a comprehensive characterization of immune infiltration in ccRCC, while the constructed model can be used effectively to predict the overall survival of ccRCC patients.
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http://dx.doi.org/10.2147/OTT.S282763DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7997590PMC
March 2021

MicroRNA-10a-3p mediates Th17/Treg cell balance and improves renal injury by inhibiting REG3A in lupus nephritis.

Int Immunopharmacol 2020 Nov 24;88:106891. Epub 2020 Aug 24.

Department of Nephrology, Henan Provincial Key Laboratory of Kidney Disease and Immunology, Henan Provincial People's Hospital (Zhengzhou University People's Hospital), Zhengzhou 450003, Henan, PR China. Electronic address:

Background: The therapeutic approaches guided toward microRNAs (miRNAs) have been extensively explored in lupus nephritis (LN), but the precise position of miR-10a-3p posted in disease is not translated thoroughly. Therein, this work pivoting on miR-10a-3p was launched with the involvement of regenerating islet-derived 3 α (REG3A).

Methods: Peripheral blood samples from LN patients and healthy controls (n = 132) were collected. miR-10a-3p and REG3A expression in peripheral blood mononuclear cells were tested. Mice were injected with miR-10a-3p agomir, miR-10a-3p antagomir and/or REG3A low expression vector for presentation of their roles in renal function, T helper cell 17 (Th17)/regulatory cell (Treg) balance, renal pathological damage, JAK2/STAT3 pathway activation and renal injury in LN. The relation between miR-10a-3p and REG3A was tested.

Results: MiR-10a-3p was down-regulated while REG3A was up-regulated in LN. Restoring miR-10a-3p or silencing REG3A decreased Th17/Treg ratio in CD4 T cells, inhibited JAK2/STAT3 pathway activation, ameliorated renal function, improved renal pathological damage and alleviated renal injury in LN. REG3A depletion negated the effects of down-regulated miR-10a-3p on LN. MiR-10a-3p targeted REG3A.

Conclusion: The work elucidates that miR-10a-3p restoration decreases Th17/Treg ratio and attenuates renal injury in LN via inhibiting REG3A and the activation of JAK2/STAT3 pathway, which renews the therapeutic reference for LN management.
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http://dx.doi.org/10.1016/j.intimp.2020.106891DOI Listing
November 2020

Mthfd2 Modulates Mitochondrial Function and DNA Repair to Maintain the Pluripotency of Mouse Stem Cells.

Stem Cell Reports 2020 08 16;15(2):529-545. Epub 2020 Jul 16.

State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China; Advanced Innovation Center for Food Nutrition and Human Health, China Agricultural University, Beijing 100083, China. Electronic address:

The pluripotency of stem cells determines their developmental potential. While the pluripotency states of pluripotent stem cells are variable and interconvertible, the mechanisms underlying the acquisition and maintenance of pluripotency remain largely elusive. Here, we identified that methylenetetrahydrofolate dehydrogenase (NAD-dependent), methenyltetrahydrofolate cyclohydrolase (Mthfd2) plays an essential role in maintaining embryonic stem cell pluripotency and promoting complete reprogramming of induced pluripotent stem cells. Mechanistically, in mitochondria, Mthfd2 maintains the integrity of the mitochondrial respiratory chain and prevents mitochondrial dysfunction. In the nucleus, Mthfd2 stabilizes the phosphorylation of EXO1 to support DNA end resection and promote homologous recombination repair. Our results revealed that Mthfd2 is a dual-function factor in determining the pluripotency of pluripotent stem cells through both mitochondrial and nuclear pathways, ultimately ensuring safe application of pluripotent stem cells.
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http://dx.doi.org/10.1016/j.stemcr.2020.06.018DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7419720PMC
August 2020

Dual-binding benzene and rhodamine B conjugate derivatives as fluorescent chemodosimeter for hypochlorite in living cell imaging.

Spectrochim Acta A Mol Biomol Spectrosc 2020 Mar 6;229:117908. Epub 2019 Dec 6.

Henan Cereal Quality and Safety Testing Key Laboratory, Institute of Quality Standards and Testing Technology for Agro-Products, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China. Electronic address:

A new probe (SRh) which based on dual-binding benzene and rhodamine B conjugate derivatives for hypochlorite detection was developed. By desulfurization effect, probe SRh displayed"Off-On" switching in its fluorogenic and chromogenic responses to hypochlorite. The detection limit of ClO was at a low level (up to 2.43 nM). Moreover, probe SRh has been applied in bioimaging with good biocompatibility and low cytotoxicity.
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http://dx.doi.org/10.1016/j.saa.2019.117908DOI Listing
March 2020

Antibiotic Application and Resistance in Swine Production in China: Current Situation and Future Perspectives.

Front Vet Sci 2019 17;6:136. Epub 2019 May 17.

Norwegian Institute of Bioeconomy Research, Ås, Norway.

To meet increasing demand for animal protein, swine have been raised in large Chinese farms widely, using antibiotics as growth promoter. However, improper use of antibiotics has caused serious environmental and health risks, in particular Antimicrobial resistance (AMR). This paper reviews the consumption of antibiotics in swine production as well as AMR and the development of novel antibiotics or alternatives in China. The estimated application of antibiotics in animal production in China accounted for about 84240 tons in 2013. Overuse and abuse of antibiotics pose a great health risk to people through food-borne antibiotic residues and selection for antibiotic resistance. China unveiled a national plan to tackle antibiotic resistance in August 2016, but more support is needed for the development of new antibiotics or alternatives like plant extracts. Antibiotic resistance has been a major global challenge, so international collaboration between China and Europe is needed.
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http://dx.doi.org/10.3389/fvets.2019.00136DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6533531PMC
May 2019

Clinical Applications of DSC-MRI Parameters Assess Angiogenesis and Differentiate Malignant From Benign Soft Tissue Tumors in Limbs.

Acad Radiol 2020 03;27(3):354-360

Department of Radiology, First Affiliated Hospital of Xi'an Jiaotong University, No. 277 Yanta West Road, Xi'an, Shaanxi 710061, P.R. China. Electronic address:

Objective: To investigate the correlation between dynamic susceptibility contrast magnetic resonance imaging (DSC-MRI) parameters and angiogenesis and to explore prospectively the feasibility of using DSC-MRI to differentiate malignant from benign soft tissue tumors (STTs) in limbs.

Methods: This prospective study included 33 patients with STTs in limbs who underwent DSC-MRI after bolus Gd-DTPA infusion. All STTs were confirmed by pathological examination after surgery and microvessel density (MVD), vascular endothelial growth factor (VEGF) expression, were evaluated by immune-histochemical analysis. Semiquantitative DSC-MRI parameters, including negative enhancement integral (NEI), maximum slopes of decrease (MSD) and increase (MSI), and mean time to enhancement were calculated by postprocessing in workstation. The correlation was analyzed between DSC-MRI parameters and angiogenesis factors. Then, the DSC-MRI parameters were compared between benign and malignant STTs and evaluated for diagnostic efficiency by receiver operating characteristic.

Results: The 33 evaluated tumors were consisted of 13 benign and 20 malignant STTs in limbs. Significant positive correlations were observed between NEI, MSD, MSI and MVD, VEGF (p < 0.05). However, mean time to enhancement had no correlation with MVD and VEGF. The benign and malignant STTs differed significantly in terms of NEI, MSD, and MSI (p < 0.05). The areas under the curve (AUC) of NEI, MSD, and MSI were 0.915, 0.862, and 0.815 for discriminating between benign and malignant STTs, respectively.

Conclusion: DSC-MRI parameters are positively correlated with MVD and VEGF, which can evaluate angiogenesis indirectly. Furthermore, DSC-MRI can be considered as one of assistant noninvasive MR imaging technique in differentiation between benign and malignant STTs in limbs.
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http://dx.doi.org/10.1016/j.acra.2019.04.023DOI Listing
March 2020

modulates pluripotency of mouse embryonic stem cells by regulating mitochondrial function and glutathione level.

Biochem J 2019 06 11;476(11):1585-1604. Epub 2019 Jun 11.

State Key Laboratories for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100094, China

Mitochondria play a central role in the maintenance of the naive state of embryonic stem cells. Many details of the mechanism remain to be fully elucidated. Solute carrier family 25 member 36 () might regulate mitochondrial function through transporting pyrimidine nucleotides for mtDNA/RNA synthesis. Its physical role in this process remains unknown; however, was recently found to be highly expressed in naive mouse embryonic stem cells (mESCs). Here, the function of was characterized as a maintenance factor of mESCs pluripotency. deficiency (via knockdown) has been demonstrated to result in mitochondrial dysfunction, which induces the differentiation of mESCs. The expression of key pluripotency markers (, , , and ) decreased, while that of key TE genes (, , and ) increased. -positive cells emerged in 6-deficient colonies under trophoblast stem cell culture conditions. As a result of 6 deficiency, mtDNA of knockdown cells declined, leading to impaired mitochondria with swollen morphology, decreased mitochondrial membrane potential, and low numbers. The key transcription regulators of mitochondrial biogenesis also decreased. These results indicate that mitochondrial dysfunction leads to an inability to support the pluripotency maintenance. Moreover, down-regulated glutathione metabolism and up-regulated focal adhesion reinforced and stabilized the process of differentiation by separately enhancing OCT4 degradation and promoting cell spread. This study improves the understanding of the function of , as well as the relationship of mitochondrial function with naive pluripotency maintenance and stem cell fate decision.
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http://dx.doi.org/10.1042/BCJ20190057DOI Listing
June 2019

Recovery of Sm(III), Co(II) and Cu(II) from waste SmCo magnet by ionic liquid-based selective precipitation process.

Waste Manag 2018 Aug 17;78:992-1000. Epub 2018 Jul 17.

CAS Key Laboratory of Design and Assembly of Functional Nanostructures, and Fujian Key Laboratory of Nanomaterials, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, Fujian 350002, PR China; Ganzhou Rare Earth Group Co., Ltd., China Southern Rare Earth, Ganzhou 341000, PR China. Electronic address:

Recovery of secondary resources including waste SmCo magnets contributes to easing the natural resources and environmental issues. For the development of recovery technologies, an ionic liquid (IL)-based precipitation route based on [trihexyl(tetradecyl)phosphonium][benzene-1,4-dioxydiacetate] ([P][BDOAC]) is presented. The slow release of [BDOAC] from [P][BDOAC] increases the kinetics difference in the formations of precipitates, and makes the precipitates contain less impurity. While cobalt-rich solution is obtained with no loss, precipitation efficiency of Sm(III) and Cu(II) reaches 96.8% and 100% respectively. Afterwards, Cu(II) is removed using aqueous ammonia and pure Cu(II) (96.4% precipitation efficiency) is obtained by adjusting the pH of cuprammonium complex solution. The proposed process without volatile diluent reveals higher separation factors of Sm/Co, Cu/Co and Sm/Cu. Lower acidities are efficient for the complete stripping of Sm(III) and Cu(II) from their precipitates. SmCl and CuCl solutions are obtained both with the high purities. Moreover, the IL-based precipitant can be regenerative. It is shown that the strategy is efficient for recovering and separating Sm, Co and Cu from the simulated leaching solution of waste SmCo magnets.
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http://dx.doi.org/10.1016/j.wasman.2018.07.004DOI Listing
August 2018

Transcriptome and metabolome analyses provide insights into root and root-released organic anion responses to phosphorus deficiency in oat.

J Exp Bot 2018 06;69(15):3759-3771

Norwegian Institute of Bioeconomy Research (NIBIO), Ås, Norway.

Roots and root-released organic anions play important roles in uptake of phosphorus (P), an essential macronutrient for food production. Oat, ranking sixth in the world's cereal production, contains valuable nutritional compounds and can withstand poor soil conditions. Our aim was to investigate root transcriptional and metabolic responses of oat grown under P-deficient and P-sufficient conditions. We conducted a hydroponic experiment and measured root morphology and organic anion exudation, and analysed changes in the transcriptome and metabolome. Oat roots showed enhanced citrate and malate exudation after 4 weeks of P deficiency. After 10 d of P deficiency, we identified 9371 differentially expressed transcripts with a 2-fold or greater change (P<0.05): 48 sequences predicted to be involved in organic anion biosynthesis and efflux were consistently up-regulated; 24 up-regulated transcripts in oat were also found to be up-regulated upon P starvation in rice and wheat under similar conditions. Phosphorylated metabolites (i.e. glucose-6-phosphate, myo-inositol phosphate) were reduced dramatically, while citrate and malate, some sugars and amino acids increased slightly in P-deficient oat roots. Our data are consistent with a strategy of increased organic anion efflux and a shift in primary metabolism in response to P deficiency in oat.
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http://dx.doi.org/10.1093/jxb/ery176DOI Listing
June 2018

Lipid Supplement in the Cultural Condition Facilitates the Porcine iPSC Derivation through cAMP/PKA/CREB Signal Pathway.

Int J Mol Sci 2018 Feb 8;19(2). Epub 2018 Feb 8.

The Animal Science and Technology College, Beijing University of Agriculture, Beijing 102206, China.

Large numbers of lipids exist in the porcine oocytes and early embryos and have the positive effects on their development, suggesting that the lipids may play an important role in pluripotency establishment and maintenance in pigs. However, the effects of lipids and their metabolites, such as fatty acids on reprogramming and the pluripotency gene expression of porcine-induced pluripotent stem cells (iPSCs), are unclear. Here, we generated the porcine iPSCs that resemble the mouse embryonic stem cells (ESCs) under lipid and fatty-acid-enriched cultural conditions (supplement of AlbuMAX). These porcine iPSCs show positive for the ESCs pluripotency markers and have the differentiation abilities to all three germ layers, and importantly, have the capability of aggregation into the inner cell mass (ICM) of porcine blastocysts. We further confirmed that lipid and fatty acid enriched condition can promote the cell proliferation and improve reprogramming efficiency by elevating cAMP levels. Interestingly, this lipids supplement promotes mesenchymal-epithelial transition (MET) through the cAMP/PKA/CREB signal pathway and upregulates the expression during porcine somatic cell reprogramming. The lipids supplement also makes a contribution to lipid droplets accumulation in the porcine iPSCs that resemble porcine preimplantation embryos. These findings may facilitate understanding of the lipid metabolism in porcine iPSCs and lay the foundation of bona fide porcine embryonic stem cell derivation.
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http://dx.doi.org/10.3390/ijms19020509DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5855731PMC
February 2018

Paracrine osteoprotegerin and β-catenin stabilization support synovial sarcomagenesis in periosteal cells.

J Clin Invest 2018 01 20;128(1):207-218. Epub 2017 Nov 20.

Departments of Orthopaedics and Oncological Sciences, and.

Synovial sarcoma (SS) is an aggressive soft-tissue sarcoma that is often discovered during adolescence and young adulthood. Despite the name, synovial sarcoma does not typically arise from a synoviocyte but instead arises in close proximity to bones. Previous work demonstrated that mice expressing the characteristic SS18-SSX fusion oncogene in myogenic factor 5-expressing (Myf5-expressing) cells develop fully penetrant sarcomagenesis, suggesting skeletal muscle progenitor cell origin. However, Myf5 is not restricted to committed myoblasts in embryos but is also expressed in multipotent mesenchymal progenitors. Here, we demonstrated that human SS and mouse tumors arising from SS18-SSX expression in the embryonic, but not postnatal, Myf5 lineage share an anatomic location that is frequently adjacent to bone. Additionally, we showed that SS can originate from periosteal cells expressing SS18-SSX alone and from preosteoblasts expressing the fusion oncogene accompanied by the added stabilization of β-catenin, which is a common secondary change in SS. Expression and secretion of the osteoclastogenesis inhibitory factor osteoprotegerin enabled early growth of SS18-SSX2-transformed cells, indicating a paracrine link between the bone and synovial sarcomagenesis. These findings explain the skeletal contact frequently observed in human SS and may provide alternate means of enabling SS18-SSX-driven oncogenesis in cells as differentiated as preosteoblasts.
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http://dx.doi.org/10.1172/JCI94955DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5749548PMC
January 2018

Tetraploid embryonic stem cells can contribute to the development of chimeric fetuses and chimeric extraembryonic tissues.

Sci Rep 2017 06 8;7(1):3030. Epub 2017 Jun 8.

State Key Laboratory for Agro biotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193, People's Republic of China.

Our study examined the in vivo chimeric and survival capacities of chimeras created by injecting tetraploid embryonic stem cells (ESCs) expressing green fluorescent protein (GFP) into diploid embryos. At 3.5 days post-coitum (dpc) and 4.5 dpc, the tetraploid ESCs were able to contribute to the inner cell mass (ICM) just as diploid ESCs tagged with GFP. At 6.5 dpc, 8.0 dpc and 10.5 dpc, the tetraploid ESCs manifested in the same location as the diploid ESCs. The GFP cells in the extraembryonic tissues and fetuses of tetraploid ESC chimeras were tetraploid as determined by fluorescence activated cell sorting (FACS). Furthermore, tetraploid ESCs contributed to the development of the placenta, embryolemma and umbilical cord at 13.5 dpc and 16.5 dpc; however, very less GFP cells were found in the fetuses of tetraploid ESC chimeras. We further found that the proliferation of tetraploid ESCs was slower than that of diploid ESCs. In addition, the relative mRNA expression in the three germ layers and the trophoblast was abnormal in the EBs of tetraploid ESCs compared with diploid ESCs. In short, slower proliferation and abnormal differentiation potential of tetraploid ESCs might be two of the reasons for their poor survival and chimeric capacities.
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http://dx.doi.org/10.1038/s41598-017-02783-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5465063PMC
June 2017

Lettuce-produced hepatitis C virus E1E2 heterodimer triggers immune responses in mice and antibody production after oral vaccination.

Plant Biotechnol J 2017 Dec 9;15(12):1611-1621. Epub 2017 Jun 9.

Institute of Biochemistry of the Romanian Academy, Bucharest, Romania.

The hepatitis C virus (HCV) is a major etiologic agent for severe liver diseases (e.g. cirrhosis, fibrosis and hepatocellular carcinoma). Approximately 140 million people have chronic HCV infections and about 500 000 die yearly from HCV-related liver pathologies. To date, there is no licensed vaccine available to prevent HCV infection and production of a HCV vaccine remains a major challenge. Here, we report the successful production of the HCV E1E2 heterodimer, an important vaccine candidate, in an edible crop (lettuce, Lactuca sativa) using Agrobacterium-mediated transient expression technology. The wild-type dimer (E1E2) and a variant without an N-glycosylation site in the E2 polypeptide (E1E2∆N6) were expressed, and appropriate N-glycosylation pattern and functionality of the E1E2 dimers were demonstrated. The humoral immune response induced by the HCV proteins was investigated in mice following oral administration of lettuce antigens with or without previous intramuscular prime with the mammalian HEK293T cell-expressed HCV dimer. Immunization by oral feeding only resulted in development of weak serum levels of anti-HCV IgM for both antigens; however, the E1E2∆N6 proteins produced higher amounts of secretory IgA, suggesting improved immunogenic properties of the N-glycosylation mutant. The mice group receiving the intramuscular injection followed by two oral boosts with the lettuce E1E2 dimer developed a systemic but also a mucosal immune response, as demonstrated by the presence of anti-HCV secretory IgA in faeces extracts. In summary, our study demonstrates the feasibility of producing complex viral antigens in lettuce, using plant transient expression technology, with great potential for future low-cost oral vaccine development.
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http://dx.doi.org/10.1111/pbi.12743DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5698045PMC
December 2017

Erratum to: Highly efficient generation of biallelic reporter gene knock-in mice via CRISPR-mediated genome editing of ESCs.

Protein Cell 2017 06;8(6):471

State Key Laboratories for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100083, China.

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http://dx.doi.org/10.1007/s13238-017-0392-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5445028PMC
June 2017

Rhizosphere Organic Anions Play a Minor Role in Improving Crop Species' Ability to Take Up Residual Phosphorus (P) in Agricultural Soils Low in P Availability.

Front Plant Sci 2016 7;7:1664. Epub 2016 Nov 7.

Division of Environment and Natural Resources, Norwegian Institute of Bioeconomy Research Ås, Norway.

Many arable lands have accumulated large reserves of residual phosphorus (P) and a relatively large proportion of soil P is less available for uptake by plants. Root released organic anions are widely documented as a key physiological strategy to enhance P availability, while limited information has been generated on the contribution of rhizosphere organic anions to P utilization by crops grown in agricultural soils that are low in available P and high in extractable Ca, Al, and Fe. We studied the role of rhizosphere organic anions in P uptake from residual P in four common crops , and in low- and high-P availability agricultural soils from long-term fertilization field trials in a mini-rhizotron experiment with four replications. Malate was generally the dominant organic anion. More rhizosphere citrate was detected in low P soils than in high P soil. showed 74-103% increase of malate in low P loam, compared with clay loam. had the greatest rhizosphere citrate concentration in all soils (5.3-15.2 μmol g root DW). also showed the highest level of root colonization by arbuscular mycorrhizal fungi (AMF; 36 and 40%), the greatest root mass ratio (0.51 and 0.66) in the low-P clay loam and loam respectively, and the greatest total P uptake (5.92 mg P/mini-rhizotron) in the low-P loam. had 15-44% more rhizosphere acid phosphatase (APase) activity, ~0.1-0.4 units lower rhizosphere pH than other species, the greatest increase in rhizosphere water-soluble P in the low-P soils, and the greatest total P uptake in the low-P clay loam. Shoot P content was mainly explained by rhizosphere APase activity, water-soluble P and pH within low P soils across species. Within species, P uptake was mainly linked to rhizosphere water soluble P, APase, and pH in low P soils. The effects of rhizosphere organic anions varied among species and they appeared to play minor roles in improving P availability and uptake.
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http://dx.doi.org/10.3389/fpls.2016.01664DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5097927PMC
November 2016

LRP5 Signaling in Osteosarcomagenesis: a Cautionary Tale of Translation from Cell Lines to Tumors.

Transl Oncol 2016 Oct;9(5):438-444

Department of Orthopaedic Surgery, University of Utah, Salt Lake City, UT, 84112; Department of Oncological Sciences, University of Utah, Salt Lake City, UT, 84112. Electronic address:

Previous reports document expression of low-density lipoprotein receptor-related protein 5 (LRP5) in osteosarcoma (OS) tissue. Expression of this Wnt receptor correlated with metastatic disease and poor disease-free survival. Forced expression of dominant-negative LRP5 (dnLRP5), which lacks the membrane binding domain of the native protein and therefore functions as a soluble receptor-sponge for Wnt ligands, reduced in vitro cellular invasion and in vivo xenograft tumor growth for osteosarcoma cell lines. Here, we use a genetically engineered mouse model of osteosarcomagenesis with and without expression of dnLRP5 to assess to what degree tumorigenesis is affected and whether Wnt/β-catenin signaling is circumvented or maintained. Each cohort of mice developed osteosarcoma at a similar ultimate prevalence, but after a slightly increased latency in those also expressing dnLRP5. On histology, there was no difference between groups, despite previous reports that the dnLRP5 osteosarcoma cells specifically undergo a mesenchymal-to-epithelial transition in vitro. Finally, immunohistochemistry showed the presence of cytosolic and nuclear β-catenin and nuclear Cyclin D1, markers consistent with preserved Wnt/β-catenin signaling despite constitutive blockade of the cell surface receipt of Wnt signaling ligand. These data suggest that canonical Wnt signaling plays a role in OS progression and that while blockade of singular nodes in signaling pathways can have dramatic effects on individual cell lines, real tumors readily evade such focused attacks.
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http://dx.doi.org/10.1016/j.tranon.2016.08.010DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5067932PMC
October 2016

Production of dengue virus envelope protein domain III-based antigens in tobacco chloroplasts using inducible and constitutive expression systems.

Plant Mol Biol 2016 Jul 26;91(4-5):497-512. Epub 2016 Apr 26.

NIBIO-Norwegian Institute of Bioeconomy Research, P.O. Box 115, 1431, Ås, Norway.

Dengue fever is a disease in many parts of the tropics and subtropics and about half the world's population is at risk of infection according to the World Health Organization. Dengue is caused by any of the four related dengue virus serotypes DEN-1, -2, -3 and -4, which are transmitted to people by Aedes aegypti mosquitoes. Currently there is only one vaccine (Dengvaxia(®)) available (limited to a few countries) on the market since 2015 after half a century's intensive efforts. Affordable and accessible vaccines against dengue are hence still urgently needed. The dengue envelop protein domain III (EDIII), which is capable of eliciting serotype-specific neutralizing antibodies, has become the focus for subunit vaccine development. To contribute to the development of an accessible and affordable dengue vaccine, in the current study we have used plant-based vaccine production systems to generate a dengue subunit vaccine candidate in tobacco. Chloroplast genome engineering was applied to express serotype-specific recombinant EDIII proteins in tobacco chloroplasts using both constitutive and ethanol-inducible expression systems. Expression of a tetravalent antigen fusion construct combining EDIII polypeptides from all four serotypes was also attempted. Transplastomic EDIII-expressing tobacco lines were obtained and homoplasmy was verified by Southern blot analysis. Northern blot analyses showed expression of EDIII antigen-encoding genes. EDIII protein accumulation levels varied for the different recombinant EDIII proteins and the different expression systems, and reached between 0.8 and 1.6 % of total cellular protein. Our study demonstrates the suitability of the chloroplast compartment as a production site for an EDIII-based vaccine candidate against dengue fever and presents a Gateway(®) plastid transformation vector for inducible transgene expression.
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http://dx.doi.org/10.1007/s11103-016-0484-5DOI Listing
July 2016

[The prevention strategy of concurrent extraction and osteoradionecrosis after radiotherapy].

Zhonghua Kou Qiang Yi Xue Za Zhi 2015 Aug;50(8):503-6

Email:

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August 2015

Highly efficient generation of biallelic reporter gene knock-in mice via CRISPR-mediated genome editing of ESCs.

Protein Cell 2016 Feb;7(2):152-6

State Key Laboratories for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100083, China.

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http://dx.doi.org/10.1007/s13238-015-0228-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4742388PMC
February 2016

[The influence of zoledronic acid on vascular endothelial cell].

Zhonghua Kou Qiang Yi Xue Za Zhi 2015 Jul;50(7):399-402

Department of Oral and Maxillofacial Surgery, School & Hospital of Stomatology, Wenzhou Medical University, Wenzhou Zhejiang 325027, China; Email:

Objective: To investigate the influence of zoledronic acid on vascular endothelial cells.

Methods: The influence of zoledronic acid on proliferation, migration and adhesion of vascular endothelial cells were tested with 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), cell migration assay and cell adhesion assay. The results of each experimental group were compared with the control group and the data statistically analyzed.

Results: In a concentration of 0-0.5 mmol/L, the absorbance value decreased from 0.09 to 0.34 as the drug concentration increased. Scratch test showed that the change of width of scratch before and after 24 hours in control, low, medium and high concentration groups were (38.7 ± 0.42), (35.8 ± 4.17), (19.9 ± 0.57) mm (P < 0.001), (12.5 ± 3.89) mm (P < 0.05). Adhesion test showed that the absorbance value in control, low, medium and high concentration groups were 1.14 ± 0.18, 0.95 ± 0.13, 0.81 ± 0.11 (P < 0.01), 0.67 ± 0.19 (P < 0.001). Comparisons between control and experimental groups were analyzed by t-test and P values < 0.05 were considered statistically significant.

Conclusions: Zoledronic acid inhibits the proliferation, migration and adhesion of vascular endothelial cells.
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July 2015

Improvement in Mouse iPSC Induction by Rab32 Reveals the Importance of Lipid Metabolism during Reprogramming.

Sci Rep 2015 Nov 12;5:16539. Epub 2015 Nov 12.

State Key Laboratories for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, 100193, China.

Induced pluripotent stem cells (iPSCs) have variable expression levels of a series of genes that affect their pluripotent potential, but the regulatory mechanisms controlling reprogramming remain unclear. By testing the efficiency of iPSC generation using Oct4, Sox2, Klf4 (termed OSK) plus one additional gene, we found that Rab32 improved reprogramming efficiency. We established a system for detecting the number and the size of lipid droplets and autophagosomes per cell for tracking their morphological changes during reprogramming. Our results showed that Rab32 increased lipid storage during the early and middle stages, and also increased autophagy during the middle stage of reprogramming. These findings were further confirmed by the up-regulation of lipid biosynthesis and autophagosome formation related genes, of which their expression could improve iPSC induction. The inhibition of lipid biosynthesis and autophagosome formation significantly reduced reprogramming efficiency, and the inhibition of lipid synthesis phenotype could be rescued by the overexpression of Rab32. In addition, the expression of pluripotency genes such as Klf2, Nr5a2 and Tbx3, was up-regulated by Rab32. These results demonstrated that Rab32 could improve the induction of iPSCs through the enhancement of lipid biosynthesis, highlighting the importance of lipid metabolism during reprogramming.
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http://dx.doi.org/10.1038/srep16539DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4642270PMC
November 2015

β-catenin stabilization enhances SS18-SSX2-driven synovial sarcomagenesis and blocks the mesenchymal to epithelial transition.

Oncotarget 2015 Sep;6(26):22758-66

Department of Orthopaedics and Huntsman Cancer Institute, University of Utah School of Medicine, Salt Lake City, UT 84112, USA.

β-catenin is a master regulator in the cellular biology of development and neoplasia. Its dysregulation is implicated as a driver of colorectal carcinogenesis and the epithelial-mesenchymal transition in other cancers. Nuclear β-catenin staining is a poor prognostic sign in synovial sarcoma, the most common soft-tissue sarcoma in adolescents and young adults. We show through genetic experiments in a mouse model that expression of a stabilized form of β-catenin greatly enhances synovial sarcomagenesis. Stabilization of β-catenin enables a stem-cell phenotype in synovial sarcoma cells, specifically blocking epithelial differentiation and driving invasion. β-catenin achieves its reprogramming in part by upregulating transcription of TCF/LEF target genes. Even though synovial sarcoma is primarily a mesenchymal neoplasm, its progression towards a more aggressive and invasive phenotype parallels the epithelial-mesenchymal transition observed in epithelial cancers, where β-catenin's transcriptional contribution includes blocking epithelial differentiation.
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http://dx.doi.org/10.18632/oncotarget.4283DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4673197PMC
September 2015

[The prevention of canine osteoradionecrosis of jaws by low-intensity ultrasound].

Zhonghua Kou Qiang Yi Xue Za Zhi 2015 May;50(5):297-301

Department of Oral and Maxillofacial Surgery, School & Hospital of Stomatology, Wenzhou Medical University, Wenzhou Zhejiang 325027, China; Email:

Objective: To investigate the preventive effect of low-intensity ultrasound on osteoradionecrosis of jaws (ORNJ).

Methods: Twenty-five canines were randomly divided into experimental group (n=20) and control group (n=5). The canines in experimental group received radiation exposure, and then were randomly subdivided into group A (n=10) and group B (n=10). Control group did not undergo radiotherapy. One month after radiotherapy, the fourth mandibular premolars of all animals were extracted. Group B was immediately treated by low-intensity ultrasound for twenty days, group A and control group did not receive any treatment. Two months after tooth extraction, the formation of ORNJ was determined and the occurrence rate of ORNJ was compared between group A and B. The microstructure of the mandible and changes in microvascular density in group A and B were evaluated and compared with those of control group.

Results: All animals in group B and group A developed ORNJ after prophylactic ultrasound was applied for twenty days. Although the imaging examination of bony density of group A and B were lower than normal animals in control group, bone density in group B was significantly better than group A. Micro-CT showed that the trabecular bone volume fraction, trabecular thickness, bone surface/bone volume and trabecular number in group B were respectively (0.187±0.029)%, (0.160±0.039) µm, (12.536±2.558)/mm, (1.227±0.192)/mm, which were all greater than group A [(0.103±0.014)%, (0.069±0.013) µm, (5.598±0.731)/mm, (0.522±0.064)/mm)] (P<0.05).

Conclusions: Although the preventive application of low intensity ultrasound can not prevent the formation of ORNJ, but can significantly improve the symptoms of ORNJ.
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May 2015

The surgical treatment of reinforced steel bar injury penetrating the skull base and maxilla-mandibular area.

J Craniofac Surg 2014 Nov;25(6):e521-3

From the *Stomatological Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, People's Republic of China; †School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou, Zhejiang, People's Republic of China; and ‡The Orthopaedic Department, the 2nd Affiliated Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, People's Republic of China.

Penetrating injuries with reinforced screwed steel bar in the skull base represent a unique challenge for oral maxillofacial surgeons. Management of these injuries is complicated by associated injuries and the proximity to vital neurovascular structures. A 35-year-old man was admitted to our hospital because of injury due to a downward fall upon a reinforced steel rod. Radiologic studies of the skull base revealed that the steel bar traversed the temporomandibular space between the left cervical spine and the mastoid process to the space between the inner side of the left mandibular ramus and the maxilla. We performed osteotomy of the left mastoid process tip and the left mandibular ramus to take out the steel bar from the maxilla and repaired the left mandible with internal fixation. Appropriate preoperative planning, including three-dimensional computed tomographic images, is integral in the surgical approach for the safe removal of such objects.
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http://dx.doi.org/10.1097/SCS.0000000000000505DOI Listing
November 2014

Transformation of cellulose and its derived carbohydrates into formic and lactic acids catalyzed by vanadyl cations.

ChemSusChem 2014 Jun 2;7(6):1557-67. Epub 2014 May 2.

State Key Laboratory of Physical Chemistry of Solid Surfaces, Collaborative Innovation Center of Chemistry for Energy Materials, National Engineering Laboratory for Green Chemical Productions of Alcohols, Ethers and Esters, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005 (PR China).

The transformation of cellulose or cellulose-derived carbohydrates into platform chemicals is the key to establish biomass-based sustainable chemical processes. The systems able to catalyze the conversion of cellulose into key chemicals in water without the consumption of hydrogen are limited. We report that simple vanadyl (VO(2+)) cations catalyze the conversions of cellulose and its monomer, glucose, into lactic acid and formic acid in water. We have discovered an interesting shift of the major product from formic acid to lactic acid on switching the reaction atmosphere from oxygen to nitrogen. Our studies suggest that VO(2+) catalyzes the isomerization of glucose to fructose, the retro-aldol fragmentation of fructose to two trioses, and the isomerization of trioses, which leads to the formation of lactic acid under anaerobic conditions. The oxidative cleavage of C-C bonds in the intermediates caused by the redox conversion of VO2(+)/VO(2+) under aerobic conditions results in formic acid and CO2. We demonstrate that the addition of an alcohol suppresses the formation of CO2 and enhances the formic acid yield significantly to 70-75 %.
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http://dx.doi.org/10.1002/cssc.201400150DOI Listing
June 2014

Chemical synthesis of lactic acid from cellulose catalysed by lead(II) ions in water.

Nat Commun 2013 ;4:2141

State Key Laboratory of Physical Chemistry of Solid Surfaces, Collaborative Innovation Center of Chemistry for Energy Materials, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China.

The direct transformation of cellulose, which is the main component of lignocellulosic biomass, into building-block chemicals is the key to establishing biomass-based sustainable chemical processes. Only limited successes have been achieved for such transformations under mild conditions. Here we report the simple and efficient chemocatalytic conversion of cellulose in water in the presence of dilute lead(II) ions, into lactic acid, which is a high-value chemical used for the production of fine chemicals and biodegradable plastics. The lactic acid yield from microcrystalline cellulose and several lignocellulose-based raw biomasses is >60% at 463 K. Both theoretical and experimental studies suggest that lead(II) in combination with water catalyses a series of cascading steps for lactic acid formation, including the isomerization of glucose formed via the hydrolysis of cellulose into fructose, the selective cleavage of the C3-C4 bond of fructose to trioses and the selective conversion of trioses into lactic acid.
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http://dx.doi.org/10.1038/ncomms3141DOI Listing
January 2014
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