Publications by authors named "Yonglin Chen"

41 Publications

Ribavirin inhibits colorectal cancer growth by downregulating PRMT5 expression and H3R8me2s and H4R3me2s accumulation.

Toxicol Appl Pharmacol 2021 03 9;415:115450. Epub 2021 Feb 9.

Department of Pharmacology, School of Basic Medical Sciences, Lanzhou University, Lanzhou 730000, People's Republic of China. Electronic address:

Eukaryotic translation initiation factor 4E (eIF4E) and protein arginine methyltransferase 5 (PRMT5) are frequently overexpressed in colorectal cancer (CRC) tissues and associated with poor prognosis. Ribavirin, the only clinically approved drug known to target eIF4E, is an anti-viral molecule currently used in hepatitis C therapy. The potential of ribavirin to treat CRC remains largely unknown. Ribavirin treatment in CRC cell lines drastically inhibited cell proliferation and colony formation, induced S phase arrest and reduced cyclin D1, cyclin A/E and proliferating cell nuclear antigen (PCNA) levels in vitro, and suppressed tumorigenesis in mouse model of colitis-associated CRC. Mechanistically, ribavirin treatment significantly reduced PRMT5 and eIF4E protein levels and the accumulation of symmetric dimethylation of histone 3 at arginine 8 (H3R8me2s) and that of histone 4 at arginine 3 (H4R3me2s). Importantly, inhibition of PRMT5 by ribavirin resulted in promoted H3R8 methylation in eIF4E promoter region. Our results demonstrate the anti-cancer efficacy of ribavirin in CRC and suggest that the anti-cancer efficacy of ribavirin may be mediated by downregulating PRMT5 levels but not its enzymatic activity.
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http://dx.doi.org/10.1016/j.taap.2021.115450DOI Listing
March 2021

MicroRNA: a novel implication for damage and protection against ionizing radiation.

Environ Sci Pollut Res Int 2021 Apr 3;28(13):15584-15596. Epub 2021 Feb 3.

Hengyang Medical College, Institute of Cytology and Genetics, The Hengyang Key Laboratory of Cellular Stress Biology, University of South China, Hengyang, 421001, Hunan Province, People's Republic of China.

Ionizing radiation (IR) is a form of high energy. It poses a serious threat to organisms, but radiotherapy is a key therapeutic strategy for various cancers. It is significant to reduce radiation injury but maximize the effect of radiotherapy. MicroRNAs (miRNAs) are posttranscriptionally regulatory factors involved in cellular radioresponse. In this review, we show how miRNAs regulate important genes on cellular response to IR-induced damage and how miRNAs participate in IR-induced carcinogenesis. Additionally, we summarize the experimental and clinical evidence for miRNA involvement in radiotherapy and discuss their potential for improvement of radiotherapy. Finally, we highlight the role that miRNAs play in accident exposure to IR or radiotherapy as predictive biomarker. miRNA therapeutics have shown great perspective in radiobiology; miRNA may become a novel strategy for damage and protection against IR.
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http://dx.doi.org/10.1007/s11356-021-12509-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7854028PMC
April 2021

Genetic variation of E6 and E7 genes of human papillomavirus 52 from Central China.

J Med Virol 2021 06 1;93(6):3849-3856. Epub 2020 Dec 1.

Department of Laboratory Medicine, Jingzhou Central Hospital, The Second Clinical Medical College, Yangtze University, Jingzhou, China.

Cervical cancer is the fourth most common malignant tumor in women worldwide and is closely related to human papillomavirus (HPV). Women have the highest susceptibility to HPV-52 type in Jingzhou, China. In this study, E6-E7 sequences of 183 HPV-52 positive samples were amplified by a polymerase chain reaction and sequenced. HPV-52 E6-E7 gene variations were analyzed. The phylogenetic tree was constructed using the Kimura 2-parameter method. The secondary structure of the protein was analyzed. The selective pressure to E6-E7 genes was estimated using PAML. In addition, the B cell epitopes of the E6-E7 sequences in HPV-52 were predicted by the ABCpred server. In E6 sequences, 15 single nucleotide variants were observed, including 6 nonsynonymous variants and 9 synonymous variants. In E7 sequences, 19 single nucleotide variants occurred, including 10 nonsynonymous variants and 9 synonymous variants. Six amino acid variants, including 3 nonconservative substitutions, were found in sequences encoding the alpha helix. Eight amino acid variants, including three nonconservative substitutions, occurred in sequences encoding the strand. Through phylogenetic analysis, the E6-E7 sequences were mainly distributed in B lineage. In HPV-52 E6-E7 sequences, no positively selected site was found. The nonconservative substitutions, such as K93R, K93E in E6, T37I, and D38N in E7, affected multiple hypothetical epitopes in the B cell. This study provides information for the investigation of HPV epidemic characters. The discovery of new variants of HPV-52 may lay the basis for the development of the virus diagnosis, further study of cervical cancer, and vaccine design in Central China.
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http://dx.doi.org/10.1002/jmv.26690DOI Listing
June 2021

iTRAQ-Based Proteomic Profiling of Potential Biomarkers in Rat Serum for Uranium Tailing Suspension Intratracheal Instillation.

J Proteome Res 2021 01 5;20(1):995-1004. Epub 2020 Nov 5.

The Hengyang Key Laboratory of Cellular Stress Biology, Key Discipline Laboratory for National Defense for Biotechnology in Uranium Mining and Hydrometallurgy, University of South China, Hengyang, Hunan 421001, P. R. China.

Protection against low-dose ionizing radiation is of great significance. Uranium tailings are formed as a byproduct of uranium mining and a potential risk to organisms. In this study, we identified potential biomarkers associated with exposure to low-dose radiation from uranium tailings. We established a Wistar rat model of low dose rate irradiation by intratracheal instillation of a uranium tailing suspension. We observed pathological changes in the liver, lung, and kidney tissues of the rats. Using isobaric tags for relative and absolute quantification, we screened 17 common differentially expressed proteins in three dose groups. We chose -1 antiproteinase (Serpina1), keratin 17 (Krt17), and aldehyde dehydrogenase (Aldh3a1) for further investigation. Our data showed that expression of Serpina1, Krt17, and Aldh3a1 had changed after the intratracheal instillation in rats, which may be potential biomarkers for uranium tailing low-dose irradiation. However, the underlying mechanisms require further investigation.
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http://dx.doi.org/10.1021/acs.jproteome.0c00766DOI Listing
January 2021

Nonintubated minimally invasive chest wall stabilization for multiple rib fractures: a prospective, single-arm study.

World J Emerg Surg 2020 09 23;15(1):53. Epub 2020 Sep 23.

Department of Thoracic Surgery, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, 200233, China.

Background: Nonintubated video-assisted thoracoscopic surgery has been widely reported in the past decade, while nonintubated chest wall stabilization has not been reported previously. The aim of this study was to evaluate the safety and feasibility of nonintubated minimally invasive chest wall stabilization in patients with multiple rib fractures.

Methods: We conducted a prospective, single-arm, observational study. In this prospective study, 20 consecutive patients with multiple rib fractures were treated using nonintubated minimally invasive chest wall stabilization.

Results: Minimally invasive chest wall stabilization was mostly performed for lateral rib fractures in this study (n = 8). The mean operation time was 92.5 min, and the mean blood loss was 49 ml. No patient required conversion to tracheal intubation. The mean extubation time of the laryngeal mask was 8.9 min; the mean postoperative fasting time was 6.1 h; the mean postoperative hospital stay was 6.2 days; the mean amount of postoperative drainage was 97.5 ml; the mean postoperative pain score was 2.9 points at 6 h, 2.8 points at 12 h, and 3.0 points at 24 h; and the mean postoperative nausea and vomiting score was 1.9 points at 6 h, 1.8 points at 12 h, and 1.7 points at 24 h.

Conclusions: Nonintubated minimally invasive chest wall stabilization is safe and feasible in carefully selected patients. Further studies with a large sample size are warranted.

Trial Registration: ChiCTR1900025698 . Registered on 5 September 2019.
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http://dx.doi.org/10.1186/s13017-020-00335-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7513522PMC
September 2020

Unveiling the Mechanism, Origin of Stereoselectivity, and Ligand-Dependent Reactivity in the Pd(II)-Catalyzed Unbiased Methylene C(sp)-H Alkenylation-Aza-Wacker Cyclization Reaction.

J Org Chem 2020 Oct 5;85(20):13191-13203. Epub 2020 Oct 5.

Department of Earth Science and Engineering, Taiyuan University of Technology, Taiyuan, Shanxi 030024, People's Republic of China.

The mechanism, origin of stereoselectivity, and ligand-dependent reactivity of Pd(II)-catalyzed methylene C(sp)-H alkenylation-aza-Wacker cyclization to form ()-β-stereogenic γ-lactam have been comprehensively studied by density functional theory (DFT) calculations. The calculated results reveal that the methylene C-H activation assisted by KCO via the concerted metalation-deprotonation mechanism is found to be the most preferred pathway, where the enantioselectivity is distinguished by the orientation of the methyl group of a substrate relative to a chiral ligand. However, the stereochemistry of the olefin moiety in the generated product is mainly determined by the oxidative addition step, where the coulombic interaction and dispersion effect differentiate the energy difference of diastereomeric transition states. In terms of the agostic interaction nature of "three-center two-electron" transition states, the discrepancy of reactivities caused by different Pd catalysts is attributed to the electron induction effect of substituents on the chiral ligands. In other words, the use of an electron-withdrawing group (e.g., -CN) in place of an electron-donating group (e.g., -OMe) enhances the oxidation state of the Pd atom and lowers vacant d orbitals of the palladium atom of the catalyst and in turn facilitates a larger amount of σ-electronic-charge injection into an empty 3d shell of the palladium center. Thus, the higher catalytic activity of the Pd catalyst with ligands substituted by an electron-withdrawing group is anticipated.
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http://dx.doi.org/10.1021/acs.joc.0c01906DOI Listing
October 2020

GSTP1 and cancer: Expression, methylation, polymorphisms and signaling (Review).

Int J Oncol 2020 Apr 10;56(4):867-878. Epub 2020 Feb 10.

Hengyang Medical College, Institute of Cytology and Genetics, Key Laboratory of Ecological Environment and Critical Human Diseases Prevention of Hunan Province Department of Education, University of South China, Hengyang, Hunan 421001, P.R. China.

Glutathione S‑transferase Pi (GSTP1) is an isozyme encoded by the GST pi gene that plays an important regulatory role in detoxification, anti‑oxidative damage, and the occurrence of various diseases. The aim of the present study was to review the association between the expression of GSTP1 and the development and treatment of various cancers, and discuss GSTP1 methylation in several malignant tumors, such as prostate, breast and lung cancer, as well as hepatocellular carcinoma; to review the association between polymorphism of the GSTP1 gene and various diseases; and to review the effects of GSTP1 on electrophilic oxidative stress, cell signal transduction, and the regulation of carcinogenic factors. Collectively, GSTP1 plays a major role in the development of various diseases.
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http://dx.doi.org/10.3892/ijo.2020.4979DOI Listing
April 2020

Prevalence and characteristics of Epstein-Barr virus associated gastric carcinoma in Gansu Province, Northwest China with mRNA expression of glycoprotein BMRF2.

J Med Virol 2020 03 2;92(3):356-363. Epub 2019 Dec 2.

School of Basic Medical Sciences, Lanzhou University, Lanzhou, Gansu, China.

Gansu province is a region with the highest gastric cancer incidence and mortality in Northwest China. Epstein-Barr virus-associated gastric carcinoma (EBVaGC) is an important subtype of gastric cancer which shows specific clinicopathological features such as older-age bias, male predominance, lower lymph-node-metastasis, and a better cancer-related survival comparing to EBV-negative gastric cancers. However, the prevalence of EBVaGC has never been studied in Gansu Province, Northwest China. The present study investigated the incidence, characteristics, and EBV messenger RNA (mRNA) profile of EBVaGC in this area. We have collected 270 stomach samples from gastric cancer patients and analyzed the presence of EBV DNA and EBV-encoded small RNAs (EBERs) by nested polymerase chain reaction (PCR) and in situ hybridization, respectively. The EBV mRNA profiling was investigated by quantitative reverse transcription PCR (qRT-PCR). EBV DNA was detected in 51/95 patients (53.7%), while EBER transcripts were detected in 18/270 patients (6.7%). EBER positivity was significantly associated with older age and less lymph node metastasis, but no obvious association with gender or histological type of tumors. The expression of EBV genes was observed with different patterns, and the mRNA of glycoprotein BMRF2 was detected in EBVaGC. The present study showed unique clinicopathological features and mRNA expression patterns of EBVaGC in Gansu Province, Northwest China, suggesting that geographic variation can contribute to new epidemiological features in EBVaGC. The transcript of glycoprotein BMRF2 was observed consistently in EBVaGC, which may serve as a biomarker and play a role in the pathogenesis of EBVaGC in Gansu Province, Northwest China.
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http://dx.doi.org/10.1002/jmv.25616DOI Listing
March 2020

Receptor for activated C kinase 1 mediates the chronic constriction injury-induced neuropathic pain in the rats' peripheral and central nervous system.

Neurosci Lett 2019 11 3;712:134477. Epub 2019 Sep 3.

Department of Anesthesiology, Affiliated Hospital of Nantong University, Nantong 226001, China. Electronic address:

Peripheral and central sensitization has been reported as significant features in the course of the occurrence and development of neuropathic pain (NP). Receptor for activated C kinase 1 (RACK1), a scaffold protein, participated in fundamental cellular activities and various neuronal functions. Peripheral and central sensitization are a state that the morphology of neuronal cell bodies as well as the corresponding function change, whether this process can be regulated by RACK1 is still unknown. In this study, the biological effects and mechanisms of RACK1 contributes to the pathogenesis of chronic constriction injury (CCI)-induced neuropathic pain were investigated. By western blot and staining, we found that RACK1 protein changed in dorsal root ganglion (DRG) neurons and spinal cord (SC) neurons except glial cells after CCI. Especially, RACK1 was co-located with IB4-, CGRP-positive neurons, suggesting it was related to integrate nociceptive information from the primary aff ;erents in DRG. The successful establishment of CCI models also directly led to mechanical allodynia and heat hyperalgesia, which could be reversed by intrathecal injection of RACK1 siRNA. Furthermore, intrathecal injection of RACK1 siRNA reduced the expression of RACK1 and accompanying spinal c-fos, which is the transcription factor and marker of neuronal activation. These results suggested that targeting RACK1 be a sensible approach for treating NP.
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http://dx.doi.org/10.1016/j.neulet.2019.134477DOI Listing
November 2019

Synthesis, toxicity and antitumor activity of cobalt carbonyl complexes targeting hepatocellular carcinoma.

Bioorg Med Chem 2019 10 23;27(20):115071. Epub 2019 Aug 23.

The First Affiliated Hospital of Lanzhou University, Lanzhou 730000, China.

Based on our previous research, a series of targeting hepatocellular carcinoma complexes, [R-Glycyrrhetinic acid-CHCH-[Co(CO)] (R = H, 1; R = NSAIDs-COOH, 2-4; R = Aromatic acid, 5-7; R = Amino acid, 8-10), were synthesized. The test showed they are slow CO releasers. Using HeLa, A549, HT-29, SMMC7721 and HepG2 cells as models, their activities against tumor cell proliferation were firstly evaluated. The resulting data show all the complexes displayed a good anti-proliferation activity against the HepG2 and SMMC-7721 liver cancer cells, and their IC values were in the range of 10.07-66.06 µM; compared with cis-platin (DDP), their activities were comparable or even better under the same condition. Among them, complexes 3, 4, 6 and 9 exhibited higher anti-proliferation activities against HepG2 and SMMC-7721 cell lines than the other cell lines. To confirm further these complexes have selectivity to the liver cells, the uptakes of complexes 3, 4, 6 and 9 by HepG2, HT-29, A549 and SMMC7721 cell lines were studied. The results show the cell uptake rates of the complexes by HepG2 cells and SMMC7721 cells were much greater than by other cells under the same condition. In following tests, the tested complexes displayed higher activities in inhibiting NF-kB, COX-2 and iNOS; and they induced HepG2 cells apoptosis by mitochondrial pathway, which assessed by staining with different fluorescent reagent DAPI, PI, Mito-Tracker Green and DCFH-DA. Meanwhile, the tested complexes up-regulated the expression levels of caspase-3 and Bax, down-regulated the Bcl-2 expression. In addition, they had no effect on zebrafish embryo survival, embryo hatching, embryonic movement, zebrafish malformation and zebrafish movement at below 0.5 µM. This suggests the complexes are potential candidates to be used in clinic for liver cancers.
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http://dx.doi.org/10.1016/j.bmc.2019.115071DOI Listing
October 2019

Digoxin improves steatohepatitis with differential involvement of liver cell subsets in mice through inhibition of PKM2 transactivation.

Am J Physiol Gastrointest Liver Physiol 2019 10 14;317(4):G387-G397. Epub 2019 Aug 14.

Section of Digestive Diseases, Yale University School of medicine, New Haven, Connecticut.

The cardiac glycoside digoxin was identified as a potent suppressor of pyruvate kinase isoform 2-hypoxia-inducible factor-α (PKM2-HIF-1α) pathway activation in liver injury mouse models via intraperitoneal injection. We have assessed the therapeutic effects of digoxin to reduce nonalcoholic steatohepatitis (NASH) by the clinically relevant oral route in mice and analyzed the cellular basis for this effect with differential involvement of liver cell subsets. C57BL/6J male mice were placed on a high-fat diet (HFD) for 10 wk and started concurrently with the gavage of digoxin (2.5, 0.5, 0.125 mg/kg twice a week) for 5 wk. Digoxin significantly reduced HFD-induced hepatic damage, steatosis, and liver inflammation across a wide dosage range. The lowest dose of digoxin (0.125 mg/kg) showed significant protective effects against liver injury and sterile inflammation. Consistently, digoxin attenuated HIF-1α sustained NLRP3 inflammasome activation in macrophages. We have reported for the first time that PKM2 is upregulated in hepatocytes with hepatic steatosis, and digoxin directly improved hepatocyte mitochondrial dysfunction and steatosis. Mechanistically, digoxin directly bound to PKM2 and inhibited PKM2 targeting HIF-1α transactivation without affecting PKM2 enzyme activation. Thus, oral digoxin showed potential to therapeutically inhibit liver injury in NASH through the regulation of PKM2-HIF-1α pathway activation with involvement of multiple cell types. Because of the large clinical experience with oral digoxin, this may have significant clinical applicability in human NASH. This study is the first to assess the therapeutic efficacy of oral digoxin on nonalcoholic steatohepatitis (NASH) in a high-fat diet (HFD) mouse model and to determine the divergent of cell type-specific effects. Oral digoxin reduced liver damage, steatosis, and inflammation in HFD mice. Digoxin attenuated hypoxia-inducible factor (HIF)-1α axis-sustained inflammasome activity in macrophages and hepatic oxidative stress response in hepatocytes via the regulation of PKM2-HIF-1α axis pathway activation. Oral digoxin may have significant clinical applicability in human NASH.
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http://dx.doi.org/10.1152/ajpgi.00054.2019DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6842989PMC
October 2019

Toxicities and beneficial protection of HS donors based on nonsteroidal anti-inflammatory drugs.

Medchemcomm 2019 May 22;10(5):742-756. Epub 2019 Mar 22.

The First Affiliated Hospital of Lanzhou University , Lanzhou , 730000 , China.

On the basis of our previous study, the HS donors based on nonsteroidal anti-inflammatory drugs (NSAIDs) were further evaluated with regard to the following aspects: animal blood and urine analyses, liver and kidney toxicities, gastrointestinal protection, anti-hypertension, and myocardial protection. The test results showed that after the successive administration of the compound for 14 days, the number of white blood cells in the blood of rats reduced, and protein and leukocytes appeared in urine; further, α-lipoic-acid-acetaminophen ester () and ibuprofen-ADTOH ester () had a certain effect on the physiological tissue and function of rat liver, and their side-effects on the kidneys were obvious. However, when compared with NSAIDs as the precursors, the tested compounds displayed much fewer side-effects; particularly, for the gastrointestinal mucosa of rats, there were hardly any side-effects. Moreover, all the three compounds decreased blood pressures in spontaneously hypertensive rats in a concentration-dependent manner, even though this antihypertensive effect was weaker than those induced by nifedipine and captopril. In addition, three compounds protected H9c2 cells from injury an antioxidation pathway; and they improved myocardial injury in spontaneously hypertensive rats. Compound is the derivative of ibuprofen and has lower toxicity to rat cardiomyocytes than ibuprofen. Therefore, it may become a better substitute of ibuprofen in patients due to its lower cardiotoxicity.
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http://dx.doi.org/10.1039/c8md00611cDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6533891PMC
May 2019

Toxicity, bioactivity, release of HS in vivo and pharmaco-kinetics of HS-donors with thiophosphamide structure.

Eur J Med Chem 2019 Aug 11;176:456-475. Epub 2019 May 11.

The First Affiliated Hospital of Lanzhou University, Lanzhou, 730000, China.

HS donors are substitutes of HS with various biological activities like inhibiting the inflammatory response and protecting myocardial cells from injury. In order to confirm whether the HS donors have drug-like properties, two series thiophosphamide HS donors were evaluated including toxicity, bioactivity and pharmacokinetic properties in vivo and in vitro. The following results were obtained. Firstly, all the compounds released HS under measuring condition; with the increase of pH value, the HS release rate of all the compounds decreased and the amount reduced, but pH value had little effect on the maximum release of HS. Secondly, in the organs and tissues of rats, the compounds released HS in the same way as in PBS. In plasma, compound 1 reached the C after administration 55 min, and no compound 1 was detected after 12 h; for compound 18, the C reached only after administration 100 min, and after 6 h, compound 18 was not detected; in organs and tissues, the HS-release rates were different from those in PBS, but the mechanism of HS release was the same. Thirdly, in the test of toxicity, all the compounds displayed low toxicities to 5 cancer cells and W138 cell lines; compounds 1 and 18 had slight effect on the physiological tissue and function of rat liver at low concentration; the compounds had almost no effect on the hatching rate, survival rate of zebrafish embryos, and the spontaneous movement of zebrafish embryos at below 0.5 μM, but when they were over 1 μM, the compounds displayed inhibitory effect in the manner of concentration dependence. Fourthly, in the course of anti-inflammatory test, all the tested compounds significantly reduced the level of TNF-α and increased the level of IL-10; when they were 100 μM, the levels of IL-10 were three times as high as those in the control group. Among them, compounds 10 and 18 displayed stronger activities than the others. In addition, the compounds protected H9c2 cells from injure and improved myocardial injury through anti-oxidation pathway. In summary, the compounds have druglike properties due to low toxicity, better activity and good pharmacokinetic property. Therefore, they have potential to be as candidates to investigate further.
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http://dx.doi.org/10.1016/j.ejmech.2019.05.017DOI Listing
August 2019

Interleukin-1 Receptor Associated Kinase 1 Mediates the Maintenance of Neuropathic Pain after Chronic Constriction Injury in Rats.

Neurochem Res 2019 May 11;44(5):1214-1227. Epub 2019 Mar 11.

Department of Anesthesiology, Affiliated Hospital of Nantong University, Nantong, 226001, China.

Neuropathic pain (NP) has complicated pathogenesis as it mainly involves a lesion or dysfunction of the somatosensory nervous system and its clinical treatment remains challenging. Chronic constriction injury (CCI) model is a widely used neuropathic pain model and involved in mechanisms including both nerve inflammatory and injury. Cytokines and their receptors play essential roles in the occurrence and persistence of neuropathic pain, but the underlying mechanisms have not well been understood. Therefore, Interleukin-1 receptor-associated kinase 1 (IRAK1) is chosen to explore the possible mechanisms of NP. In the present study, IRAK1 was found to persistently increase in the dorsal root ganglion (DRG) and spinal cord (SC) during CCI detected by western blot. The staining further confirmed that IRAK1 was mainly co-located in the DRG astrocytes or SC neurons, but less in the DRG microglia or SC astrocytes. Moreover, the region of increased IRAK1 expression was observed in superficial laminae of the spinal dorsal horn, which was the nociceptive neuronal expression domain, suggesting that IRAK1 may mediated CCI-induced pain by nociceptive primary afferent. In addition, intrathecal injection of Toll-like receptor 4 (TLR4) inhibitor or IRAK1 siRNA decreased the expression of IRAK1 accompanied with the alleviation of CCI-induced neuropathic pain. The upregulation of p-NF-κB expression was reversed by IRAK1 siRNA in SC, and intrathecal injection of p-NF-κB inhibitor relieved neuropathic pain. Taking together, targeting IRAK1 may be a potential treatment for chronic neuropathic pain.
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http://dx.doi.org/10.1007/s11064-019-02767-8DOI Listing
May 2019

Chemokine CXCL10/CXCR3 signaling contributes to neuropathic pain in spinal cord and dorsal root ganglia after chronic constriction injury in rats.

Neurosci Lett 2019 02 15;694:20-28. Epub 2018 Nov 15.

Department of Anesthesiology, Affiliated Hospital of Nantong University, Nantong 226001, China. Electronic address:

Inflammatory cytokines and chemokines play essential roles in the occurrence and persistence of neuropathic pain (NP). Chronic constriction injury (CCI) enhances the activation of p-ERK, which is involved in neuropathic pain. Although the chemokine CXCL10 and its receptor CXCR3 are implicated in the pathophysiology of itch, it is largely unexplored for neuropathic pain. In this study, we determined the role of the CXCL10-CXCR3 axis in NP using a well-established CCI model. CCI significantly induced mechanical allodynia and thermal hyperalgesia. Following the pain course, a significant increase of CXCL10 and CXCR3 in both dorsal root ganglion (DRG) neurons and spinal cord (SC) neurons was detected in rats. Furthermore, intrathecal injection of CXCR3 inhibitor AMG487 was found to attenuate pain hypersensitivity in a dose-dependent manner in CCI. The expression of p-ERK was also depressed after intrathecal injection of AMG487 associated with a significant laxation of hyperalgesia, which demonstrated that the interaction of CXCL10/CXCR3 possibly took part in neuropathic pain by regulating p-ERK signaling in the SC. Overall, these findings demonstrate that the CXCL10/CXCR3 signaling pathway is critical in CCI.
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http://dx.doi.org/10.1016/j.neulet.2018.11.021DOI Listing
February 2019

Translocation Associated Membrane Protein 1 Contributes to Chronic Constriction Injury-Induced Neuropathic Pain in the Dorsal Root Ganglion and Spinal Cord in Rats.

J Mol Neurosci 2018 Dec 18;66(4):535-546. Epub 2018 Oct 18.

Department of Anesthesiology, Affiliated Hospital of Nantong University, Nantong, 226001, China.

Neuropathic pain is a severe debilitating state caused by injury or dysfunction of somatosensory nervous system, and the clinical treatment is still challenging. Translocation associated membrane protein 1 (TRAM1), an adapter protein, participates in a variety of transduction pathways and mediates the biological functions such as cell proliferation, activation, and differentiation. However, whether TRAM1 is involved in the pathogenesis of neuropathic pain is still unclear. In our study, we reported the role of TRAM1 in the maintenance of neuropathic pain induced by chronic constriction injury (CCI) on rats. By western blot and staining, we found that TRAM1 increased in the dorsal root ganglion (DRG) neurons and spinal cord (SC) neurons after CCI. Being similar to IB4-, CGRP-positive expressed area, TRAM1 also expressed in the superficial laminae of the spinal cord dorsal horn (SCDH), suggesting it was related to the innervations of the primary afferents. Moreover, intrathecal injection of TRAM1 siRNA or Toll-like receptor 4 (TLR4) inhibitor induced low expression of TRAM1 in SC, which alleviated the pain response induced by CCI. The upregulation of p-NF-κB expression was reversed by TRAM1 siRNA in SC and DRG, and intrathecal injection of p-NF-κB inhibitor relieved neuropathic pain. All the data indicated that TRAM1 could take part in CCI-induced pain and might be a potential treatment for chronic neuropathic pain.
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http://dx.doi.org/10.1007/s12031-018-1187-yDOI Listing
December 2018

Syntheses and anti-cancer activity of CO-releasing molecules with targeting galactose receptors.

Org Biomol Chem 2018 11;16(43):8115-8129

Institute of Medicinal Chemistry, School of Pharmacy of Lanzhou University, Lanzhou 730000, China.

CO-releasing molecules (CORMs) containing cobalt have many bioactivities, but most of them do not dissolve in water and have no selectivity to tissue and organs. On the basis of the specific recognition of galactose or sialic acid by a receptor, a series of CORMs based on carbohydrates were synthesized and evaluated. The test results show that all the complexes displayed anticancer activity. Among them, the effects of the complexes of galactose (1), GalNAc (8) and sialic acid (10) were very distinct. Complex 1 displayed higher activity against HeLa, HePG2, MCF-7 and HT-29 cell proliferation than cis-platin (DDP), and its selectivity was far much better than DDP compared with normal cell W138. Furthermore, the uptakes of complexes 1, 8 and 10 by HePG2, HT-29, A549 and RAW264.7 cell lines were studied. The uptake ratio of each cell line for complex 1 was different, and the order of uptake ratio in the four cell lines was HePG2 > HT-29 > RAW264.7 > A549. The HePG2 cells absorbed complex 1 beyond 60% after incubation for 8 h, while A549 absorbed only 27.8%. For complex 8, the uptake trend was similar to that of complex 1 with it being absorbed by all the four cancer cells, but the uptake rate was lower. However, differently, complex 10 was absorbed heavily by macrophage RAW264.7, followed by HePG2; after 8 h incubation, the uptake ratio of RAW264.7 was over 50%. In addition, the mechanism of action was explored, and the results showed that the complexes inhibited cell cycle arrest at the G2/M phase; complex 1 up-regulated the expression levels of caspase-3 and Bax, and down-regulated the Bcl-2 expression, giving rise to HePG2 cell apoptosis.
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http://dx.doi.org/10.1039/c8ob01921eDOI Listing
November 2018

β-Hydroxybutyrate protects from alcohol-induced liver injury via a Hcar2-cAMP dependent pathway.

J Hepatol 2018 09 27;69(3):687-696. Epub 2018 Apr 27.

Section of Digestive Diseases, Yale University School of Medicine, New Haven, CT 06520, USA; USA West Haven Veterans Medical Center, West Haven, CT 06516, USA. Electronic address:

Background & Aims: Sterile inflammation resulting in alcoholic hepatitis (AH) occurs unpredictably after many years of excess alcohol intake. The factors responsible for the development of AH are not known but mitochondrial damage with loss of mitochondrial function are common features. Hcar2 is a G-protein coupled receptor which is activated by β-hydroxybutyrate (BHB). We aimed to determine the relevance of the BHB-Hcar2 pathway in alcoholic liver disease.

Methods: We tested if loss of BHB production can result in increased liver inflammation. We further tested if BHB supplementation is protective in AH through interaction with Hcar2, and analyzed the immune and cellular basis for protection.

Results: Humans with AH have reduced hepatic BHB, and inhibition of BHB production in mice aggravated ethanol-induced AH, with higher plasma alanine aminotransferase levels, increased steatosis and greater neutrophil influx. Conversely supplementation of BHB had the opposite effects with reduced alanine aminotransferase levels, reduced steatosis and neutrophil influx. This therapeutic effect of BHB is dependent on the receptor Hcar2. BHB treatment increased liver Il10 transcripts, and promoted the M2 phenotype of intrahepatic macrophages. BHB also increased the transcriptional level of M2 related genes in vitro bone marrow derived macrophages. This skewing towards M2 related genes is dependent on lower mitochondrial membrane potential (Δψ) induced by BHB.

Conclusions: Collectively, our data shows that BHB production during excess alcohol consumption has an anti-inflammatory and hepatoprotective role through an Hcar2 dependent pathway. This introduces the concept of metabolite-based therapy for AH.

Lay Summary: Alcoholic hepatitis is a life-threatening condition with no approved therapy that occurs unexpectedly in people who consume excess alcohol. The liver makes many metabolites, and we demonstrate that loss of one such metabolite β-hydroxybutyrate occurs in patients with alcoholic hepatitis. This loss can increase alcohol-induced liver injury, and β-hydroxybutyrate can protect from alcohol-induced liver injury via a receptor on liver macrophages. This opens the possibility of metabolite-based therapy for alcoholic hepatitis.
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http://dx.doi.org/10.1016/j.jhep.2018.04.004DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6098974PMC
September 2018

The D5 region of the intelectin domain is a new type of carbohydrate recognition domain in the intelectin gene family.

Dev Comp Immunol 2018 08 3;85:150-160. Epub 2018 Apr 3.

Marine Biotechnology Research Center, Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, Institute of Developmental Biology, School of Life Sciences, Shandong University, Jinan, Shandong, China. Electronic address:

Intelectin is a recently characterized soluble galactofuranose-binding lectin that exists in species ranging from amphioxus to human. Interestingly, intelectin does not contain a canonical carbohydrate-recognition domain (CRD). Therefore, we designed serial deletions of intelectin in the Chinese amphioxus (Branchiostoma belcheri tsingtauense, AmphiITLN71469) in order to identify functional regions required for carbohydrate binding. Our results revealed that Domain 5 (aa 203-302) was able to bind lipopolysaccarides (LPS) or peptidoglycan (PGN) and agglutinate bacteria as efficiently as the full-length protein. Three dimensional (3D) atomic models of Domain 5 were generated by ab initio based program QUARK and by Iterative Threading Assembly Refinement (I-TASSER) programs, in which four amino acids mediating calcium-binding (G54-G55-G56-E91) were identified by hemagglutination assay. Furthermore, a striking functional conservation of Domain 5 was detected in zebrafish intelectin 1. Taken together, our findings identified for the first time a new CRD domain in intelectin, thereby providing new knowledge leading to a better understanding of pathogen-host interactions.
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http://dx.doi.org/10.1016/j.dci.2018.02.021DOI Listing
August 2018

Digoxin Suppresses Pyruvate Kinase M2-Promoted HIF-1α Transactivation in Steatohepatitis.

Cell Metab 2018 02;27(2):339-350.e3

Section of Digestive Diseases, Yale University, New Haven, CT 06520, USA; West Haven Veterans Medical Center, West Haven, CT 06516, USA. Electronic address:

Sterile inflammation after tissue damage is a ubiquitous response, yet it has the highest amplitude in the liver. This has major clinical consequences, for alcoholic and non-alcoholic steatohepatitis (ASH and NASH) account for the majority of liver disease in industrialized countries and both lack therapy. Requirements for sustained sterile inflammation include increased oxidative stress and activation of the HIF-1α signaling pathway. We demonstrate the ability of digoxin, a cardiac glycoside, to protect from liver inflammation and damage in ASH and NASH. Digoxin was effective in maintaining cellular redox homeostasis and suppressing HIF-1α pathway activation. A proteomic screen revealed that digoxin binds pyruvate kinase M2 (PKM2), and independently of PKM2 kinase activity results in chromatin remodeling and downregulation of HIF-1α transactivation. These data identify PKM2 as a mediator and therapeutic target for regulating liver sterile inflammation, and demonstrate a novel role for digoxin that can effectively protect the liver from ASH and NASH.
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http://dx.doi.org/10.1016/j.cmet.2018.01.007DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5806149PMC
February 2018

PKM2 is involved in neuropathic pain by regulating ERK and STAT3 activation in rat spinal cord.

J Headache Pain 2018 Jan 18;19(1). Epub 2018 Jan 18.

Department of Pathogen Biology, Medical College, Nantong University, Nantong, Jiangsu, 2266001, China.

Background: Pyruvate kinase isozymes M2 (PKM2), as a member of pyruvate kinase family, plays a role of glycolytic enzyme in glucose metabolism. It also functions as protein kinase in cell proliferation, signaling, immunity, and gene transcription. In this study, the role of PKM2 in neuropathic pain induced by chronic constriction injury (CCI) was investigated.

Methods: Rats were randomly grouped to establish CCI models. PKM2, extracellular regulated protein kinases (EKR), p-ERK, signal transducers and activators of transcription (STAT3), p-STAT3, phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) and p-PI3K/AKT proteins expression in spinal cord was examined by Western blot analysis. Cellular location of PKM2 was examined by immunofluorescence. Knockdown of PKM2 was achieved by intrathecal injection of specific small interfering RNA (siRNA). Von Frey filaments and radiant heat tests were performed to determine mechanical allodynia and thermal hyperalgesia respectively. Lactate and adenosine triphosphate (ATP) contents were measured by specific kits. Tumor necrosis factor alpha (TNF-α) and interleukin-1 beta (IL-1β) levels were detected by ELISA kits.

Results: CCI markedly increased PKM2 level in rat spinal cord. Double immunofluorescent staining showed that PKM2 co-localized with neuron, astrocyte, and microglia. Intrathecal injection of PKM2 siRNA not only attenuated CCI-induced ERK and STAT3 activation, but also attenuated mechanical allodynia and thermal hyperalgesia induced by CCI. However, PKM2 siRNA failed to inhibit the activation of AKT. In addition, PKM2 siRNA significantly suppressed the production of lactate and pro-inflammatory mediators.

Conclusion: Our findings demonstrate that inhibiting PKM2 expression effectively attenuates CCI-induced neuropathic pain and inflammatory responses in rats, possibly through regulating ERK and STAT3 signaling pathway.
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http://dx.doi.org/10.1186/s10194-018-0836-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5773456PMC
January 2018

Extracellular Mitochondrial DNA Is Generated by Fibroblasts and Predicts Death in Idiopathic Pulmonary Fibrosis.

Am J Respir Crit Care Med 2017 12;196(12):1571-1581

4 Department of Pathology, Stony Brook University School of Medicine, Stony Brook, New York.

Rationale: Idiopathic pulmonary fibrosis (IPF) involves the accumulation of α-smooth muscle actin-expressing myofibroblasts arising from interactions with soluble mediators such as transforming growth factor-β1 (TGF-β1) and mechanical influences such as local tissue stiffness. Whereas IPF fibroblasts are enriched for aerobic glycolysis and innate immune receptor activation, innate immune ligands related to mitochondrial injury, such as extracellular mitochondrial DNA (mtDNA), have not been identified in IPF.

Objectives: We aimed to define an association between mtDNA and fibroblast responses in IPF.

Methods: We evaluated the response of normal human lung fibroblasts (NHLFs) to stimulation with mtDNA and determined whether the glycolytic reprogramming that occurs in response to TGF-β1 stimulation and direct contact with stiff substrates, and spontaneously in IPF fibroblasts, is associated with excessive levels of mtDNA. We measured mtDNA concentrations in bronchoalveolar lavage (BAL) from subjects with and without IPF, as well as in plasma samples from two longitudinal IPF cohorts and demographically matched control subjects.

Measurements And Main Results: Exposure to mtDNA augments α-smooth muscle actin expression in NHLFs. The metabolic changes in NHLFs that are induced by interactions with TGF-β1 or stiff hydrogels are accompanied by the accumulation of extracellular mtDNA. These findings replicate the spontaneous phenotype of IPF fibroblasts. mtDNA concentrations are increased in IPF BAL and plasma, and in the latter compartment, they display robust associations with disease progression and reduced event-free survival.

Conclusions: These findings demonstrate a previously unrecognized and highly novel connection between metabolic reprogramming, mtDNA, fibroblast activation, and clinical outcomes that provides new insight into IPF.
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http://dx.doi.org/10.1164/rccm.201612-2480OCDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5754440PMC
December 2017

Bile acids initiate cholestatic liver injury by triggering a hepatocyte-specific inflammatory response.

JCI Insight 2017 03 9;2(5):e90780. Epub 2017 Mar 9.

Department of Internal Medicine and Yale Liver Center, Yale University School of Medicine, New Haven, Connecticut, USA.

Mechanisms of bile acid-induced (BA-induced) liver injury in cholestasis are controversial, limiting development of new therapies. We examined how BAs initiate liver injury using isolated liver cells from humans and mice and in-vivo mouse models. At pathophysiologic concentrations, BAs induced proinflammatory cytokine expression in mouse and human hepatocytes, but not in nonparenchymal cells or cholangiocytes. These hepatocyte-specific cytokines stimulated neutrophil chemotaxis. Inflammatory injury was mitigated in mice treated with BA or after bile duct ligation, where less hepatic infiltration of neutrophils was detected. Neutrophils in periportal areas of livers from cholestatic patients also correlated with elevations in their serum aminotransferases. This liver-specific inflammatory response required BA entry into hepatocytes via basolateral transporter Ntcp. Pathophysiologic levels of BAs induced markers of ER stress and mitochondrial damage in mouse hepatocytes. Chemokine induction by BAs was reduced in hepatocytes from mice, while liver injury was diminished both in conventional and hepatocyte-specific mice, confirming a role for Tlr9 in BA-induced liver injury. These findings reveal potentially novel mechanisms whereby BAs elicit a hepatocyte-specific cytokine-induced inflammatory liver injury that involves innate immunity and point to likely novel pathways for treating cholestatic liver disease.
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http://dx.doi.org/10.1172/jci.insight.90780DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5333973PMC
March 2017

Structure analysis of a heteropolysaccharide from Taraxacum mongolicum Hand.-Mazz. and anticomplementary activity of its sulfated derivatives.

Carbohydr Polym 2016 Nov 29;152:241-252. Epub 2016 Jun 29.

The MOE Key Laboratory for Standardization of Chinese Medicines and the SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Road, Shanghai 201203, PR China. Electronic address:

A homogenous water-soluble polysaccharide, DPSW-A, with a deduced chemical structure was extracted from the herb Taraxacum mongolicum Hand.-Mazz. Moreover, 80.813-kDa DPSW-A is composed of three types of monosaccharide, namely rhamnose, arabinose, and galactose, at a molar ratio of 1.0:10.7:11.9. The main chain of DPSW-A contains Terminal-Galp, 1,3-Galp, 1,6-Galp, 1,3,6-Galp, and 1,2,4-Rhap; the branched chain contains Terminal-Araf, 1,5-Araf, and 1,3,5-Araf. The sulfated derivatives prepared from DPSW-A showed inhibitory effects on complement activation through the classical pathway (CH50: Sul-DPSW-A, 3.94±0.43μg/mL; heparin, 104.40±3.82μg/mL) and alternative pathway (AP50: Sul-DPSW-A, 42.76±0.46μg/mL; heparin, 43.42±0.22μg/mL). Mechanism studies indicated that Sul-DPSW-A inhibited complement activation by blocking C1q, C1r, C1s, and C9, but not C2, C3, C4, and C5. In addition, Sul-DPSW-A displayed limited anticoagulant effects. These results suggest that Sul-DPSW-A prepared from DPSW-A is valuable for treating diseases caused by excessive complement system activation.
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http://dx.doi.org/10.1016/j.carbpol.2016.06.110DOI Listing
November 2016

Syntheses, properties and bio-activities of water-soluble CO-releasing molecule based on manganese.

J Biol Inorg Chem 2016 10 27;21(7):807-24. Epub 2016 Jul 27.

School of Stomatology, Lanzhou University, Lanzhou, 730000, China.

A series of water-soluble CO-releasing molecules, [Mn(CO)3NH2CHRCO2]2 (1-3), [M(CO)3Br[(Py-C = N)(Gly) n CO2] (M = Mn, Re, 4-7), Mn(CO)4[S2CNC m H n CO2] (8-12), were synthesized and characterized by (1)H NMR, IR and ESI-HRMS. The stability of all the complexes in solution was evaluated by means of UV, IR and (1)H NMR. Among all the complexes, complex 4 and complex 8 were stable in H2O, acidic aqueous solution and basic media; complex 1 was stable in acidic aqueous solution and weak basic media (pH < 9.4). The assays showed that each complex has CO-release ability; excess sodium dithionite can enhance CO release. Among them, complexes 8-12 were fast CO-releasers. In the test of the cell proliferation, all the complexes showed anti-proliferative activities for HeLa and HepG2. In particular, complex 8 displayed a 3.5-fold anti-proliferative activity on HeLa cells (IC50 23.13 μM) and fivefold on HepG2 cells (34.00 μM) compared with 5-FU. What is more, the complexes distinctly influenced cell cycle and promoted cell apoptosis; complex 1 arrested HeLa cells in S phase, whereas complex 4 and complex 8 arrested in G2/M phase; all the complexes induced HeLa cells "Early apoptosis". In addition, all complexes 1, 4 and 8 decreased intracellular nitrite levels, and complex 8 was stronger than both of the others. All these data demonstrate that complex 8 has potential to be a drug candidate. Three different categories of water-soluble CORMs 1-12 were synthesized, and their stability were evaluated. The biological activities were preliminarily evaluated. This includes anti-proliferation and anti-inflammatory properties.
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http://dx.doi.org/10.1007/s00775-016-1379-2DOI Listing
October 2016

Toxicity, bio-distribution and metabolism of CO-releasing molecules based on cobalt.

Free Radic Biol Med 2016 08 30;97:362-374. Epub 2016 Jun 30.

School of Stomatology of Lanzhou University, Lanzhou 730000, China.

CO-releasing molecules (CORMs) containing [Co2(CO)6] moiety show many bioactivities, such as anti-inflammatory and antitumor cell proliferation. However, so far, no one knows their properties in vivo. So, here, we evaluated some these kind CORMs from drug-like properties including cytotoxicity, toxicity in vivo, distribution and metabolism. The results show all the tested complexes displayed antiproliferative activity to HeLa cell and HepG2 cell lines, and their IC50 values were 36-110µM against HeLa cells and 39-140µM against HepG2 cells. Toxicity tests of mice, we used oral acute toxic class method and got their LD50 values; among them, LD50 of complex 1 and complex 4 were in 2500-5000mgkg(-1) and complex 7 over 5000mgkg(-1). The developmental toxicities of the complexes were investigated in embryonic zebrafish. The mortality, hatch rate, malformation, heart rate, spontaneous movement, and larval behavior were examined, and we found both complexes 4 and 7 have not toxicity at low concentration (<1.0μM) but have higher toxicity at high concentration (>5.0μM). After several consecutive i.p administrations, tested complexes severely damaged rat liver and kidney in both functional and morphological aspects. Through metal ion measurement using ICP-AES, we found the tested complexes were unevenly distributed in tissues and organs; complex 4 has a big prone to collect in liver, whereas complex 7 easily enters to kidney. After administration 480min later, most of complex 7 excreted from kidney and entered urine, while complex 4 needed 9h at least. This results show cobalt did not accumulate, and could excrete with the urine. In vivo, Co(0) in complexes was oxidised to Co(II). In addition, the substituents significantly affected the rate of CO-release, cytotoxicity and their bio-distribution. In the view of these aspects, the CORMs based cobalt has a potential property to be a medicine.
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http://dx.doi.org/10.1016/j.freeradbiomed.2016.06.029DOI Listing
August 2016

A Homogeneous Polysaccharide from Fructus Schisandra chinensis (Turz.) Baill Induces Mitochondrial Apoptosis through the Hsp90/AKT Signalling Pathway in HepG2 Cells.

Int J Mol Sci 2016 Jun 28;17(7). Epub 2016 Jun 28.

The MOE Key Laboratory for Standardization of Chinese Medicines, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.

According to the potential anti-hepatoma therapeutic effect of Schisandra chinensis polysaccharides presented in previous studies, a bioactive constituent, homogeneous Schisandra chinensis polysaccharide-0-1 (SCP-0-1), molecular weight (MW) circa 69.980 kDa, was isolated and purified. We assessed the efficacy of SCP-0-1 against human hepatocellular liver carcinoma (HepG2) cells to investigate the effects of its antitumour activity and molecular mechanisms. Anticancer activity was evaluated using microscopy, 3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyltetrazolium bromide (MTT) assay, Hoechst 33258 staining, acridine orange (AO) staining, flow cytometry (FCM), and cell-cycle analysis. SCP-0-1 inhibited the HepG2 cells' growth via inducing apoptosis and second gap/mitosis (G2/M) arrest dose-dependently, with a half maximal inhibitory concentration (IC50) value of 479.63 µg/mL. Western blotting of key proteins revealed the apoptotic and autophagic potential of SCP-0-1. Besides, SCP-0-1 upregulated Bcl-2 Associated X Protein (Bax) and downregulated B-cell leukemia/lymphoma 2 (Bcl-2) in the HepG2 cells. The expression of caspase-3, -8, and -9; poly (ADP-ribose) polymerase (PARP); cytochrome c (Cyt C); tumor protein 53 (p53); survivin; sequestosome 1 (p62); microtubule-associated protein 1 light chain-3B (LC3B); mitogen-activated protein kinase p38 (p38); extracellular regulated protein kinases (ERK); c-Jun N-terminal kinase (JNK); protein kinase B (AKT); and heat shock protein 90 (Hsp90) were evaluated using Western blotting. Our findings demonstrate a novel mechanism through which SCP-0-1 exerts its antiproliferative activity and induces mitochondrial apoptosis rather than autophagy. The induction of mitochondrial apoptosis was attributed to the inhibition of the Hsp90/AKT signalling pathway in an extracellular signal-regulated kinase-independent manner. The results also provide initial evidence on a molecular basis that SCP-0-1 can be used as an anti-hepatocellular carcinoma therapeutic agent in the future.
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http://dx.doi.org/10.3390/ijms17071015DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4964391PMC
June 2016

Hepatocyte mitochondrial DNA drives nonalcoholic steatohepatitis by activation of TLR9.

J Clin Invest 2016 Mar 25;126(3):859-64. Epub 2016 Jan 25.

Nonalcoholic steatohepatitis (NASH) is the most common liver disease in industrialized countries. NASH is a progressive disease that can lead to cirrhosis, cancer, and death, and there are currently no approved therapies. The development of NASH in animal models requires intact TLR9, but how the TLR9 pathway is activated in NASH is not clear. Our objectives in this study were to identify NASH-associated ligands for TLR9, establish the cellular requirement for TLR9, and evaluate the role of obesity-induced changes in TLR9 pathway activation. We demonstrated that plasma from mice and patients with NASH contains high levels of mitochondrial DNA (mtDNA) and intact mitochondria and has the ability to activate TLR9. Most of the plasma mtDNA was contained in microparticles (MPs) of hepatocyte origin, and removal of these MPs from plasma resulted in a substantial decrease in TLR9 activation capacity. In mice, NASH development in response to a high-fat diet required TLR9 on lysozyme-expressing cells, and a clinically applicable TLR9 antagonist blocked the development of NASH when given prophylactically and therapeutically. These data demonstrate that activation of the TLR9 pathway provides a link between the key metabolic and inflammatory phenotypes in NASH.
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http://dx.doi.org/10.1172/JCI83885DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4767345PMC
March 2016

Neutral monosaccharide composition analysis of plant-derived oligo- and polysaccharides by high performance liquid chromatography.

Carbohydr Polym 2016 Jan 22;136:1273-80. Epub 2015 Oct 22.

The MOE Key Laboratory for Standardization of Chinese Medicines and the SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Road, Shanghai 201203, PR China. Electronic address:

A novel analytical method for neutral monosaccharide composition analysis of plant-derived oligo- and polysaccharides was developed using hydrophilic interaction liquid chromatography coupled to a charged aerosol detector. The effects of column type, additives, pH and column temperature on retention and separation were evaluated. Additionally, the method could distinguish potential impurities in samples, including chloride, sulfate and sodium, from sugars. The results of validation demonstrated that this method had good linearity (R(2) ≥ 0.9981), high precision (relative standard deviation ≤ 4.43%), and adequate accuracy (94.02-103.37% recovery) and sensitivity (detection limit: 15-40 ng). Finally, the monosaccharide compositions of the polysaccharide from Eclipta prostrasta L. and stachyose were successfully profiled through this method. This report represents the first time that all of these common monosaccharides could be well-separated and determined simultaneously by high performance liquid chromatography without additional derivatization. This newly developed method is convenient, efficient and reliable for monosaccharide analysis.
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http://dx.doi.org/10.1016/j.carbpol.2015.10.028DOI Listing
January 2016
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