Publications by authors named "Bo-Wen Li"

46 Publications

Correction to: Galectin-9 promotes a suppressive microenvironment in human cancer by enhancing STING degradation.

Oncogenesis 2022 Jan 19;11(1). Epub 2022 Jan 19.

Department of Biotherapy, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangdong Key Laboratory of Nasopharyngeal Carcinoma Diagnosis and Therapy, Sun Yat-sen University Cancer Center, and School of Life Sciences, Sun Yat-sen University, 510060, Guangzhou, P. R. China.

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http://dx.doi.org/10.1038/s41389-021-00375-2DOI Listing
January 2022

Effect of glucose metabolism disorders on the short-term prognosis in neonates with asphyxia: a multicenter study in Hubei Province, China.

Zhongguo Dang Dai Er Ke Za Zhi 2021 Dec;23(12):1208-1213

School of Medicine, Wuhan University of Science and Technology, Wuhan 430065, China.

Objectives: To study the effect of glucose metabolism disorders on the short-term prognosis in neonates with asphyxia.

Methods: A retrospective analysis was performed on the medical data of the neonates with asphyxia who were admitted to 52 hospitals in Hubei Province of China from January to December, 2018 and had blood glucose data within 12 hours after birth. Their blood glucose data at 1, 2, 6, and 12 hours after birth (with an allowable time error of 0.5 hour) were recorded. According to the presence or absence of brain injury and/or death during hospitalization, the neonates were divided into a poor prognosis group with 693 neonates and a good prognosis group with 779 neonates. The two groups were compared in the incidence of glucose metabolism disorders within 12 hours after birth and short-term prognosis.

Results: Compared with the good prognosis group, the poor prognosis group had a significantly higher proportion of neonates from secondary hospitals (48.5% vs 42.6%, <0.05) or with severe asphyxia (19.8% vs 8.1%, <0.05) or hypothermia therapy (4.8% vs 1.5%, <0.05), as well as a significantly higher incidence rate of disorder of glucose metabolism (18.8% vs 12.5%, <0.05). Compared with the good prognosis group, the poor prognosis group had a significantly higher incidence rate of disorder of glucose metabolism at 1, 2, and 6 hours after birth (<0.05). The multivariate logistic regression analysis showed that recurrent hyperglycemia (adjusted odds ratio=2.380, 95% confidence interval: 1.275-4.442, <0.05) was an independent risk factor for poor prognosis in neonates with asphyxia.

Conclusions: Recurrent hyperglycemia in neonates with asphyxia may suggest poor short-term prognosis, and it is necessary to strengthen the early monitoring and management of the nervous system in such neonates.
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http://dx.doi.org/10.7499/j.issn.1008-8830.2108188DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8690707PMC
December 2021

Enhanced Separation Efficiency and Purity of Circulating Tumor Cells Based on the Combined Effects of Double Sheath Fluids and Inertial Focusing.

Front Bioeng Biotechnol 2021 27;9:750444. Epub 2021 Oct 27.

Department of Laboratory Medicine, Daping Hospital, Army Medical University, Chongqing, China.

Circulating tumor cells (CTCs) play a crucial role in solid tumor metastasis, but obtaining high purity and viability CTCs is a challenging task due to their rarity. Although various works using spiral microchannels to isolate CTCs have been reported, the sorting purity of CTCs has not been significantly improved. Herein, we developed a novel double spiral microchannel for efficient separation and enrichment of intact and high-purity CTCs based on the combined effects of two-stage inertial focusing and particle deflection. Particle deflection relies on the second sheath to produce a deflection of the focused sample flow segment at the end of the first-stage microchannel, allowing larger particles to remain focused and entered the second-stage microchannel while smaller particles moved into the first waste channel. The deflection of the focused sample flow segment was visualized. Testing by a binary mixture of 10.4 and 16.5 μm fluorescent microspheres, it showed 16.5 μm with separation efficiency of 98% and purity of 90% under the second sheath flow rate of 700 μl min. In biological experiments, the average purity of spiked CTCs was 74% at a high throughput of 1.5 × 10 cells min, and the recovery was more than 91%. Compared to the control group, the viability of separated cells was 99%. Finally, we validated the performance of the double spiral microchannel using clinical cancer blood samples. CTCs with a concentration of 2-28 counts ml were separated from all 12 patients' peripheral blood. Thus, our device could be a robust and label-free liquid biopsy platform in inertial microfluidics for successful application in clinical trials.
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http://dx.doi.org/10.3389/fbioe.2021.750444DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8578950PMC
October 2021

Microglia polarization in ischemic stroke: complex mechanisms and therapeutic interventions.

Chin Med J (Engl) 2021 Sep 15;134(20):2415-2417. Epub 2021 Sep 15.

Department of Neurosurgery, The First People's Hospital of Changzhou, Changzhou, Jiangsu 213003, China.

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http://dx.doi.org/10.1097/CM9.0000000000001711DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8654435PMC
September 2021

The F-box E3 ubiquitin ligase AtSDR is involved in salt and drought stress responses in Arabidopsis.

Gene 2022 Jan 14;809:146011. Epub 2021 Oct 14.

College of Biological Sciences and Engineering, Jiangxi Agricultural University, Nanchang 330045, PR China. Electronic address:

F-box protein genes have been shown to play vital roles in plant development and stress respones. In Arabidopsis, there are more than 600 F-box proteins, and most of their functions are unclear. The present study shows that the F-box (SKP1-Cullin/CDC53-F-box) gene At5g15710 (Salt and Drought Responsiveness, SDR) is involved in abiotic stress responses in Arabidopsis. SDR is expressed in all tissues of Arabidopsis and is upregulated by salt and heat stresses and ABA treatment but downregulated by drought stress. Subcellular localization analysis shows that the SDR protein colocalizes with the nucleus. 35S:AntiSDR plants are hypersensitive to salt stress, but 35S:SDR plants display a salt-tolerant phenotype. Furthermore, 35S:SDR plants are hypersensitive to drought stress, while 35S:AntiSDR plants are significantly more drought tolerant. Overall, our results suggest that SDR is involved in salt and drought stress responses in Arabidopsis.
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http://dx.doi.org/10.1016/j.gene.2021.146011DOI Listing
January 2022

Development and validation of a prognostic nomogram for colorectal cancer after surgery.

World J Clin Cases 2021 Jul;9(21):5860-5872

Department of Gastroenterology, The Frist Hospital of China Medical University, Shenyang 110000, Liaoning Province, China.

Background: A nomogram is a diagram that aggregates various predictive factors through multivariate regression analysis, which can be used to predict patient outcomes intuitively. Lymph node (LN) metastasis and tumor deposit (TD) conditions are two critical factors that affect the prognosis of patients with colorectal cancer (CRC) after surgery. At present, few effective tools have been established to predict the overall survival (OS) of CRC patients after surgery.

Aim: To screen out suitable risk factors and to develop a nomogram that predicts the postoperative OS of CRC patients.

Methods: Data from a total of 3139 patients diagnosed with CRC who underwent surgical removal of tumors and LN resection from 2010 to 2015 were collected from the Surveillance, Epidemiology, and End Results program. The data were divided into a training set ( = 2092) and a validation set ( = 1047) at random. The Harrell concordance index (C-index), Akaike information criterion (AIC), and area under the curve (AUC) were used to assess the predictive performance of the N stage from the American Joint Committee Cancer tumor-node-metastasis classification, LN ratio (LNR), and log odds of positive lymph nodes (LODDS). Univariate and multivariate analyses were utilized to screen out the risk factors significantly correlating with OS. The construction of the nomogram was based on Cox regression analysis. The C-index, receiver operating characteristic (ROC) curve, and calibration curve were employed to evaluate the discrimination and prediction abilities of the model. The likelihood ratio test was used to compare the sensitivity and specificity of the final model to the model with the N stage alone to evaluate LN metastasis.

Results: The predictive efficacy of the LODDS was better than that of the LNR based on the C-index, AIC values, and AUC values of the ROC curve. Seven independent predictive factors, namely, race, age at diagnosis, T stage, M stage, LODDS, TD condition, and serum carcinoembryonic antigen level, were included in the nomogram. The C-index of the nomogram for OS prediction was 0.8002 (95%CI: 0.7839-0.8165) in the training set and 0.7864 (95%CI: 0.7604-0.8124) in the validation set. The AUC values of the ROC curve predicting the 1-, 3-, and 5-year OS were 0.846, 0.841, and 0.825, respectively, in the training set and 0.823, 0.817, and 0.835, respectively, in the validation test. Great consistency between the predicted and actual observed OS for the 1-, 3-, and 5-year OS in the training set and validation set was shown in the calibration curves. The final nomogram showed a better sensitivity and specificity than the nomogram with N stage alone for evaluating LN metastasis in both the training set (-4668.0 -4688.3, < 0.001) and the validation set (-1919.5 -1919.8, < 0.001) through the likelihood ratio test.

Conclusion: The nomogram incorporating LODDS, TD, and other risk factors showed great predictive accuracy and better sensitivity and specificity and represents a potential tool for therapeutic decision-making.
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http://dx.doi.org/10.12998/wjcc.v9.i21.5860DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8316929PMC
July 2021

Identification and Validation of Plasma Metabolomic Signatures in Precancerous Gastric Lesions That Progress to Cancer.

JAMA Netw Open 2021 06 1;4(6):e2114186. Epub 2021 Jun 1.

Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Department of Cancer Epidemiology, Peking University Cancer Hospital & Institute, Beijing, China.

Importance: Metabolic deregulation plays an important role in gastric cancer (GC) development. To date, no studies have comprehensively explored the metabolomic profiles along the cascade of gastric lesions toward GC.

Objective: To draw a metabolic landscape and define metabolomic signatures associated with the progression of gastric lesions and risk of early GC.

Design, Setting, And Participants: A 2-stage, population-based cohort study was initiated in 2017 in Linqu County, Shandong Province, China, a high-risk area for GC. Prospective follow-up was conducted during the validation stage (June 20, 2017, to May 27, 2020). A total of 400 individuals were included based on the National Upper Gastrointestinal Cancer Early Detection Program in China. The discovery stage involved 200 individuals with different gastric lesions or GC (high-grade intraepithelial neoplasia or invasive GC). The validation stage prospectively enrolled 152 individuals with gastric lesions who were followed up for 118 to 1063 days and 48 individuals with GC.

Exposures: Metabolomic profiles and metabolite signatures were examined based on untargeted plasma metabolomics assay.

Main Outcomes And Measures: The risk of GC overall and early GC (high-grade intraepithelial neoplasia), and progression of gastric lesions.

Results: Of the 400 participants, 124 of 200 (62.0%) in the discovery set were men; mean (SD) age was 56.8 (7.5) years. In the validation set, 136 of 200 (68.0%) were men; mean (SD) age was 57.5 (8.1) years. Distinct metabolomic profiles were noted for gastric lesions and GC. Six metabolites, including α-linolenic acid, linoleic acid, palmitic acid, arachidonic acid, sn-1 lysophosphatidylcholine (LysoPC)(18:3), and sn-2 LysoPC(20:3) were significantly inversely associated with risk of GC overall and early GC (high-grade intraepithelial neoplasia). Among these metabolites, the first 3 were significantly inversely associated with gastric lesion progression, especially for the progression of intestinal metaplasia (α-linolenic acid: OR, 0.42; 95% CI, 0.18-0.98; linoleic acid: OR, 0.43; 95% CI, 0.19-1.00; and palmitic acid: OR, 0.32; 95% CI, 0.13-0.78). Compared with models including only age, sex, Helicobacter pylori infection, and gastric histopathologic findings, integrating these metabolites significantly improved the performance for predicting the progression of gastric lesions (area under the curve [AUC], 0.86; 95% CI, 0.70-1.00 vs AUC, 0.69; 95% CI, 0.50-0.88; P = .02) and risk of early GC (AUC, 0.83; 95% CI, 0.58-1.00 vs AUC, 0.61; 95% CI, 0.31-0.91; P = .03).

Conclusions And Relevance: This study defined metabolite signatures that might serve as meaningful biomarkers for assessing high-risk populations and early diagnosis of GC, possibly advancing targeted GC prevention and control.
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http://dx.doi.org/10.1001/jamanetworkopen.2021.14186DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8220475PMC
June 2021

[Research progress of the role of IGF-1 in metabolic diseases and the effect of exercise intervention].

Sheng Li Xue Bao 2021 Apr;73(2):342-352

Department of Sports and Cardiovascular Health, Institute of Sports Biology, College of Physical Education, Shaanxi Normal University, Xi'an 710119, China.

Insulin-like growth factor-1 (IGF-1) is a peptide with a similar molecular structure to insulin. IGF-1 plays a key role in tissue growth and development, as well as cell metabolism, proliferation, differentiation and apoptosis. Liver is the main source of IGF-1, with the production of IGF-1 up to 75% of the total in the whole body, while the remaining 25% are secreted by skeletal muscles, heart, kidney, spleen and other organs. Target organs of IGF-1 include heart, blood vessels, liver, bone and skeletal muscles. It has been well documented that IGF-1 plays an important role in the prevention and treatment of metabolic diseases. Different types of exercise have different effects on IGF-1 expression with organ differences. In this article, we reviewed the preventive and therapeutic effects of IGF-1 on metabolic diseases and IGF-1-mediated exercise-induced benefits.
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April 2021

A novel antitumor peptide inhibits proliferation and migration and promotes apoptosis in glioma cells by regulating the MKK6/p38 signaling pathway.

Neoplasma 2021 Jul 13;68(4):732-741. Epub 2021 Apr 13.

Department of Neurosurgery, The First People's Hospital of Changzhou, Changzhou, Jiangsu, China.

Protein- or peptide-based therapeutics have emerged as an innovative strategy for the treatment of cancer. Our previous research demonstrated that tripartite motif 9 short isoform (TRIM9s) is a tumor suppressor in glioma. In this report, we investigated whether a new peptide derived from TRIM9s, named T9sP, inhibits glioma progression and determined the possible molecular mechanism. The CCK-8 proliferation assay was performed in LN229 and U251 glioma cells. The scratch-wound assay was used to determine the migration of the cells. Apoptosis was assessed by flow cytometry using Annexin V-FITC/PI double staining method. The relative protein expression levels were detected by immunoblot analysis. The cell-penetrating peptide TAT was fused with T9sP to form TAT-T9sP. TAT-T9sP efficiently penetrated through the cell membrane of both LN229 and U251 cells. TAT-T9sP inhibited proliferation and migration and promoted apoptosis of glioma cells. TAT-T9sP activated p38 signaling by upregulating MKK6, and a p38 inhibitor, SB203580, reversed the inhibitory effects of TAT-T9sP on glioma cells. These results indicated the potential of TAT-T9sP for the development of a new anti-glioma medicine.
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http://dx.doi.org/10.4149/neo_2021_201109N1196DOI Listing
July 2021

Infant attraction: why social bridging matters for female leadership in Tibetan macaques.

Curr Zool 2020 Dec 13;66(6):635-642. Epub 2020 Jun 13.

School of Resources and Environmental Engineering, Anhui University, Hefei 230601, China.

Leadership is a key issue in the study of collective behavior in social animals. Affiliation-leadership models predict that dyadic partner preferences based on grooming relationships or alliance formation positively affect an individual's decision to follow or support a conspecific. In the case of many primate species, females without young infants are attracted to mother-infant dyads. However, the effects of mother-infant-female associations on affiliation-leadership models remain less clear. In free-ranging Tibetan macaques , we used social network analysis to examine the importance of "mother-infant-adult female" social bridging events as a predictor of who leads and who follows during group movement. Social bridging is a common behavior in Tibetan macaques and occurs when 2 adults, generally females, engage in coordinated infant handling. Using eigenvector centrality coefficients of social bridging as a measure of social affiliation, we found that among lactating females, initiating bridging behavior with another female played a significant role in leadership success, with the assisting female following the mother during group movement. Among nonlactating females, this was not the case. Our results indicate that infant attraction can be a strong trigger in collective action and directing group movement in Tibetan macaques and provides benefits to mothers who require helpers and social support in order to ensure the safety of their infants. Our study provides new insights into the importance of the third-party effect in rethinking affiliation-leadership models in group-living animals.
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http://dx.doi.org/10.1093/cz/zoaa026DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7769585PMC
December 2020

Rapid Fabrication of Protein Microarrays via Autogeneration and on-Chip Purification of Biotinylated Probes.

ACS Synth Biol 2020 09 20;9(9):2267-2273. Epub 2020 Aug 20.

Key Laboratory of Biopesticide and Chemical Biology of Education Ministry, Key Laboratory of Pathogenic Fungi and Mycotoxins of Fujian Province, School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, 350002, P. R. China.

A streamlined approach toward the rapid fabrication of streptavidin-biotin-based protein microarrays was investigated. First, using our engineered versatile plasmid (pBADcM-tBirA) and an optimal coexpression strategy for biotin ligase and biotin acceptor peptide (BAP) chimeric recombinant protein, an autogeneration system for biotinylated probes was developed. This system permitted an advantageous biotinylation of BAP chimeric recombinant proteins, providing a strategy for the high-throughput synthesis of biotinylated probes. Then, to bypass the conventional rate-limiting steps, we employed an on-chip purification process to immobilize the biotinylated probes with high-throughput recombinant lysates. The integration of the autogeneration of probes and on-chip purification not only contributed to the effective and reliable fabrication of the protein microarray, but also enabled simplification of the process and an automated throughput format. This labor- and cost-effective approach may facilitate the use of protein microarrays for diagnosis, pharmacology, proteomics, and other laboratory initiatives.
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http://dx.doi.org/10.1021/acssynbio.0c00343DOI Listing
September 2020

Galectin-9 promotes a suppressive microenvironment in human cancer by enhancing STING degradation.

Oncogenesis 2020 Jul 6;9(7):65. Epub 2020 Jul 6.

Department of Biotherapy, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangdong Key Laboratory of Nasopharyngeal Carcinoma Diagnosis and Therapy, Sun Yat-sen University Cancer Center, and School of Life Sciences, Sun Yat-sen University, 510060, Guangzhou, P. R. China.

Galectin-9 (Gal-9) is known to enhance the expansion of myeloid-derived suppressor cells (MDSCs) in murine models. Its contribution to the expansion of MDSCs in human malignancies remain to be investigated. We here report that Gal-9 expression in nasopharyngeal carcinoma (NPC) cells enhances the generation of MDSCs (CD33CD11bHLA-DR) from CD33 bystander cells. The underlying mechanisms involve both the intracellular and secreted Gal-9. Inside carcinoma cells, Gal-9 up-regulates the expression of a variety of pro-inflammatory cytokines which are critical for MDSC differentiation, including IL-1β and IL-6. This effect is mediated by accelerated STING protein degradation resulting from direct interaction of the Gal-9 carbohydrate recognition domain 1 with the STING C-terminus and subsequent enhancement of the E3 ubiquitin ligase TRIM29-mediated K48-linked ubiquitination of STING. Moreover, we showed that extracellular Gal-9 secreted by carcinoma cells can enter the myeloid cells and trigger the same signaling cascade. Consistently, high concentrations of tumor and plasma Gal-9 are associated with shortened survival of NPC patients. Our findings unearth that Gal-9 induces myeloid lineage-mediated immunosuppression in tumor microenvironments by suppressing STING signaling.
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http://dx.doi.org/10.1038/s41389-020-00248-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7338349PMC
July 2020

Effectiveness and safety of the use of antifibrinolytic agents in total-knee arthroplasty: A meta-analysis.

Medicine (Baltimore) 2020 May;99(20):e20214

Department of Spinal Surgery, The Third Affiliated Hospital of Anhui Medical University, Hefei.

Background: Antifibrinolytic agents have been successfully used to reduce blood transfusion demand in patients undergoing elective knee arthroplasty. The purpose of this study was to investigate different antifibrinolytic agents for patients undergoing total-knee arthroplasty (TKA).

Methods: We searched the randomized controlled trials assessing the effect of antifibrinolytic agents on TKA in MEDLINE, PubMed, Embase, and the Cochrane Library. Participants are divided into antifibrinolytic agent group and control group under TKA. Double extraction technology is used and the quality of its methodology is evaluated before analysis. Outcomes analyzed included blood loss, number of blood transfusions, rates of blood transfusion, and deep vein thrombosis (DVT).

Results: A total of 28 randomized controlled trials involving 1899 patients were included in this study. Compared with the control group, the antifibrinolytic agents group exhibited significantly reduced the amounts of total blood loss (weighted mean difference [WMD] with 95% confidence interval [CI]: -272.19, -338.25 to -206.4), postoperative blood loss (WMD with 95% CI: -102.83, -157.64 to -46.02), average units of blood transfusion (risk ratio with 95% CI: 0.7, 0.12 to 0.24), and average blood transfusion volumes (WMD with 95% CI: -1.34, -1.47 to -1,21). Antifibrinolytic agents significantly reduced the rate of blood transfusions and did not increase the occurrence risk of intraoperative blood loss and DVT. Several limitations should also be acknowledged such as the heterogeneity among the studies.

Conclusion: The application of antifibrinolytic agents can significantly reduce blood loss and blood transfusion requirements. Additionally, these agents did not increase the risk of DVT in patients undergoing TKAs.
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http://dx.doi.org/10.1097/MD.0000000000020214DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7254857PMC
May 2020

Av3 Single-Stranded DNA Aptamer Attenuates Vascular Smooth Muscle Cell Proliferation and Migration via Ras-PI3K/MAPK Pathway.

Cardiovasc Ther 2020 21;2020:6869856. Epub 2020 Jan 21.

Department of Cardiovascular Surgery, Renmin Hospital of Wuhan University, Wuhan, China.

Objectives: To observe the effect of av3 single-stranded (ss) DNA on proliferation and migration of vascular smooth muscle cells (VSMCs) and its potential mechanism.

Background: Percutaneous transluminal coronary angioplasty (PTCA) is currently the preferred method for the treatment of coronary heart disease. However, vascular restenosis still occurs after PTCA treatment, severely affecting the clinical efficacy of PTCA. Integrin av3 single-stranded (ss) DNA on proliferation and migration of vascular smooth muscle cells (VSMCs) and its potential mechanism.

Methods: In this experiment, we used systematic evolution of ligands by exponential enrichment (SELEX) to screen out av3 single-stranded (ss) DNA on proliferation and migration of vascular smooth muscle cells (VSMCs) and its potential mechanism. 3 single-stranded (ss) DNA on proliferation and migration of vascular smooth muscle cells (VSMCs) and its potential mechanism. 3 single-stranded (ss) DNA on proliferation and migration of vascular smooth muscle cells (VSMCs) and its potential mechanism. 3 single-stranded (ss) DNA on proliferation and migration of vascular smooth muscle cells (VSMCs) and its potential mechanism.

Results: In the present study, we found that av3 single-stranded (ss) DNA on proliferation and migration of vascular smooth muscle cells (VSMCs) and its potential mechanism. < 0.05). Av3 single-stranded (ss) DNA on proliferation and migration of vascular smooth muscle cells (VSMCs) and its potential mechanism. < 0.05). Av < 0.05). Av.

Conclusions: The findings suggest that av3 ssDNA inhibited the proliferation and migration of VSMCs by suppressing the activation of Ras-PI3K/MAPK signaling.3 single-stranded (ss) DNA on proliferation and migration of vascular smooth muscle cells (VSMCs) and its potential mechanism.
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http://dx.doi.org/10.1155/2020/6869856DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6995496PMC
June 2020

Avβ3 single-stranded DNA aptamer attenuates vascular restenosis via Ras-PI3K/MAPK pathway in rats after percutaneous transluminal coronary angioplasty.

Artif Organs 2020 Jun 27;44(6):611-619. Epub 2020 Jan 27.

Department of Cardiovascular Surgery, Renmin Hospital of Wuhan University, Wuhan, China.

Our aim was to investigate the effect of avβ3 single-stranded DNA aptamer (avβ3 ssDNA) on vascular restenosis in rats after percutaneous transluminal coronary angioplasty (PTCA) via the Ras-PI3K/MAPK pathway. Sixty Sprague-Dawley rats were randomly divided into six groups: sham-operated, PTCA, PTCA+cilengitide (18 mg/kg, n = 8), and avβ3 ssDNA treatment at 50, 100, and 200 μg/kg. Hematoxylin-eosin staining was performed to evaluate the successful establishment of the PTCA model and to assess the degree of intimal hyperplasia. Immunofluorescence and in situ hybridization were carried out to observe the level of avβ3. Immunohistochemistry was used to detect the expression of E-cadherin, N-cadherin, α-smooth muscle actin (α-SMA), angiotensin 1 (ANG1), and ANG2. The expression of osteopontin (OPN), focal adhesion kinase (FAK), Ras, mitogen-activated protein kinase (MAPK), phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K), signal transducer and activator of transcription 1 (STAT1), and GTPase was observed by the western blot and quantitative reverse transcription polymerase chain reaction. Compared with rats subjected to PTCA only, those treated with avβ3 ssDNA showed significantly decreased vascular occlusion rate (P < .05). The protein expression of avβ3, OPN, p-FAK, ANG2, and E-cadherin was significantly increased by avβ3 ssDNA (P < .05), while the levels of ANG1, α-SMA, N-cadherin Ras, MAPK, PI3K, STAT1, and GTPase were significantly decreased (P < .05). Avβ3 ssDNA reduced the proliferation, migration, epithelial-mesenchymal transition, and vascular remodeling of vascular smooth muscle cells, and the mechanism may be related to the Ras-PI3K/MAPK pathway.
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http://dx.doi.org/10.1111/aor.13622DOI Listing
June 2020

A study to evaluate herb-drug interaction underlying mechanisms: An investigation of ginsenosides attenuating the effect of warfarin on cardiovascular diseases.

Eur J Pharm Sci 2020 Jan 25;142:105100. Epub 2019 Oct 25.

Fujian Provincial Key Laboratory of Cancer Metastasis Chemoprevention and Chemotherapy,College of Chemistry, Fuzhou University, Fuzhou, 350116, China; Marine Drug R&D Center, Institute of Oceanography, Minjiang University, Fuzhou, 350108, China. Electronic address:

Warfarin and ginseng have been widely used in the treatment of cardiovascular diseases. However, the clinical safety and effectiveness of herb-drug combination treatment are still controversial. Therefore, it is very essential to probe the interaction between warfarin and ginseng. In this study, in vitro and in vivo study was carried out to demonstrate that whether there is an interaction between warfarin and ginsenosides (GS), which is the main component of ginseng. In vitro study showed that the adhesion ability between endothelial cells and matrigel/platelets was enhanced due to the up-regulating expression of intercellular adhesion molecule (ICAM-1) and vascular cell adhesion molecule (VCAM-1) proteins by treatment of warfarin+GS combination compared to warfarin/GS treatment alone. Moreover, GS could weaken the anticoagulation effect of warfarin in hyperlipemia rats owning to the increased expression levels of coagulation factors and hepatic cytochrome P450 enzymes in plasma after long-term co-administration of warfarin with GS. The results of both in vitro and in vivo study demonstrated that there is a serious interaction between warfarin and ginseng, which may deteriorate atherosclerosis and thrombosis after combined use of warfarin and GS.
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http://dx.doi.org/10.1016/j.ejps.2019.105100DOI Listing
January 2020

A thienopyridine, CB-20, exerts diuretic activity by inhibiting urea transporters.

Acta Pharmacol Sin 2020 Jan 18;41(1):65-72. Epub 2019 Jun 18.

State Key Laboratory of Natural and Biomimetic Drugs, Department of Pharmacology, School of Basic Medical Sciences, Peking University, Beijing, 100191, China.

Urea transporters (UTs) are transmembrane proteins selectively permeable to urea and play an important role in urine concentration. UT-knockout mice exhibit the urea-selective urine-concentrating defect, without affecting electrolyte balance, suggesting that UT-B inhibitors have the potential to be developed as novel diuretics. In this study, we characterized a novel compound 5-ethyl-2-methyl-3-amino-6-methylthieno[2,3-b]pyridine-2,5-dicarboxylate (CB-20) with UT inhibitory activity as novel diuretics with excellent pharmacological properties. This compound was discovered based on high-throughput virtual screening combined with the erythrocyte osmotic lysis assay. Selectivity of UT inhibitors was assayed using transwell chambers. Diuretic activity of the compound was examined in rats and mice using metabolic cages. Pharmacokinetic parameters were detected in rats using LC-MS/MS. Molecular docking was employed to predict the potential binding modes for the CB-20 with human UT-B. This compound dose-dependently inhibited UT-facilitated urea transport with IC values at low micromolar levels. It exhibited nearly equal inhibitory activity on both UT-A1 and UT-B. After subcutaneous administration of CB-20, the animals showed polyuria, without electrolyte imbalance and abnormal metabolism. CB-20 possessed a good absorption and rapid clearance in rat plasma. Administration of CB-20 for 5 days did not cause significant morphological abnormality in kidney or liver tissues of rats. Molecular docking showed that CB-20 was positioned near several residues in human UT-B, including Leu364, Val367, and so on. This study provides proof of evidence for the prominent diuretic activity of CB-20 by specifically inhibiting UTs. CB-20 or thienopyridine analogs may be developed as novel diuretics.
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http://dx.doi.org/10.1038/s41401-019-0245-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7468274PMC
January 2020

TCF21 inhibits tumor-associated angiogenesis and suppresses the growth of cholangiocarcinoma by targeting PI3K/Akt and ERK signaling.

Am J Physiol Gastrointest Liver Physiol 2019 06 28;316(6):G763-G773. Epub 2019 Mar 28.

Department of Oncology, Hunan Provincial People's Hospital and the First Affiliated Hospital of Hunan Normal University , Changsha , People's Republic of China; and Key Laboratory of Study and Discovery of Small Targeted Molecules of Hunan Province, Changsha, Hunan, People's Republic of China.

Tumor-associated angiogenesis plays a critical role in the pathogenesis of cholangiocarcinoma (CCA). In this study, we examined the biological effects and molecular mechanisms of transcription factor 21 (TCF21) on CCA-associated angiogenesis. TCF21 expression was compared between 15 pairs of peritumor normal tissues and CCA tissues and also between normal bile duct epithelial cells and two CCA cell lines (QBC-939 and TFK-1) using real-time PCR and Western blot. With the use of both CCA cell lines as the model system, we stably expressed TCF21 by lentiviral transduction (Lv-TCF21). In vivo, we monitored xenograft growth from different CCA cells, measured tumor-associated angiogenesis by histological analysis, and determined the expressions and circulatory levels of VEGFA and PDGF-BB by immunohistochemistry and ELISA, respectively. In vitro, we assessed the effects of conditioned medium collected from different CCA cells on the viability, migration, and tube formation of endothelial cells and explored the significance of phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt), as well as ERK1/2 signaling in this process. TCF21 was significantly downregulated in CCA tissues or cell lines. Ectopic expression of TCF21 in CCA cells inhibited xenograft growth or tumor-associated angiogenesis in vivo and targeted the expression and secretion of proangiogenic factors, VEGFA and PDGF-BB. In vitro, the conditioned medium collected from Lv-TCF21 CCA cells significantly reduced the viability, migration, and tube formation of endothelial cells. On the molecular level, the targeting of PI3K/Akt and ERK1/2 signaling mediated the anti-angiogenic activity of TCF21. TCF21 presents growth-inhibitory and anti-angiogenic activities, and thus the elevation of TCF21 expression may provide therapeutic benefits for CCA. Transcription factor 21 (TCF21) is downregulated in cholangiocarcinoma (CCA) tissues or cells. TCF21 inhibits the growth of xenografts derived from CCA cells. TCF21 suppresses in vivo tumor-associated angiogenesis. TCF21 targets expression and production of proangiogenic factors from CCA cells. The targeting of phosphatidylinositol 3-kinase/protein kinase B and ERK1/2 signaling mediates the anti-angiogenesis of TCF21.
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http://dx.doi.org/10.1152/ajpgi.00264.2018DOI Listing
June 2019

Improvement of Mechanical Stability for Single Unit Recording Based on Skull Cap in Living Chinchilla.

Curr Med Sci 2019 Feb 13;39(1):166-172. Epub 2019 Mar 13.

Department of Otolaryngology-Head & Neck Surgery, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, 710061, China.

Three-point head fixation was constructed to provide mechanical stability for single unit recording (SUR) on vestibular sensory system in living chinchilla previously. However, it is no more qualified to this work when the stimulation intensity becomes large because of frequent unit losing and neuron damage, which strongly implies that the mechanical stability has been broken during the stimulation. Here, we constructed a novel head fixation (skull cap assistant head fixation) provided by skull cap on the basis of three-point head fixation in order to improve the mechanical stability for SUR under the stimulation with large magnitude. The large area bone connection is the feature and advantage of this improved method, which directly fixes the tested local nervous tissue and microelectrode in an intact stable system through skull cap except two ear bars and a tube face mask. Our data exhibited that skull cap assistant head fixation could significantly improve the success rate of neural response activity recording in the population of semicircular canal neurons under the stimulation with large intensity (amplitude ≥100 deg/s). Based on the analysis of neural response activity and noise base-line during stimulation, our data further indicated that this method could significantly improve the mechanical stability for SUR during high-speed motion stimulation on vestibular system in living chinchilla. Skull cap assistant head fixation extends the application of SUR on vestibular neuron in linear response range and provides a solid foundation for electrophysiological research on vestibular sensory system in further studies.
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http://dx.doi.org/10.1007/s11596-019-2015-5DOI Listing
February 2019

Pd/Cu-Catalyzed Cascade C(sp)-H Arylation and Intramolecular C-N Coupling: A One-Pot Synthesis of 3,4-2 H-Quinolinone Skeletons.

Org Lett 2019 03 25;21(6):1668-1671. Epub 2019 Feb 25.

College of Life and Environment Science , §MUC Center on Translational Neuroscience , and ¶Experimental Center of Chemistry , Minzu University of China , Beijing 100081 , China.

In this letter, we successfully explored a cascade Pd/Cu-catalyzed intermolecular C(sp)-H arylation of amides and intramolecular C-N coupling reaction. This method provides a one-pot strategy to synthesize 3,4-2 H-quinolinone with good regioselectivity of C-H arylation and C-N coupling from C-I and C-X bonds from readily available starting materials.
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http://dx.doi.org/10.1021/acs.orglett.9b00214DOI Listing
March 2019

Circular RNA Expression in Oral Squamous Cell Carcinoma.

Front Oncol 2018 8;8:398. Epub 2018 Oct 8.

Department of Oral and Maxillofacial Surgery, Peking University Shenzhen Hospital, Shenzhen, China.

Circular RNA (circRNA) is a type of non-coding RNA molecule that affects the cellular regulatory network by sequestering microRNA (miRNA) like a sponge. This study was performed to identify differentially-expressed circRNA in oral squamous cell carcinomas (OSCCs). By high-throughput sequencing, microarray circRNA expression profiles were acquired from patients with OSCCs ( = 8) and controls ( = 8), which totaled 1921 existing circRNA molecules and 10021 novel circRNA molecules. Most of the circular RNA is from exons and distributed in the No. 1 and 2 chromosomes. Eight up-regulated and down-regulated circRNA molecules were identified as differentially-expressed in OSCCs. Among this, the expression of circ_000334, circ_006740, and circ_006371 are significantly down-regulated in 42 pairs of samples, which means that these circRNA molecules might be implicated in oncogenesis and development of OSCCs.
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http://dx.doi.org/10.3389/fonc.2018.00398DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6187971PMC
October 2018

Orychophragines A-C, Three Biologically Active Alkaloids from Orychophragmus violaceus.

Org Lett 2018 02 17;20(3):656-659. Epub 2018 Jan 17.

Beijing Institute of Radiation Medicine , Beijing 100850, People's Republic of China.

Orychophragines A-C (1-3), three new alkaloids with an novel 2-piperazinone-fused 2,4-dioxohexahydro-1,3,5-triazine skeleton, were isolated from the seeds of Orychophragmus violaceus. Their structures were established on the basis of spectroscopic analysis and X-ray crystallographic analysis. Orychophragines A (1) exhibited remarkable cytotoxicity against HepG2, A549, Hela, and HCT-116 cells with IC values of 7.73, 10.79, 11.91, and 9.93 μM, respectively. Orychophragines C (3) showed moderate Co γ radiation protection activity in HUVEC cells. A plausible biosynthetic pathway for 1-3 was proposed.
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http://dx.doi.org/10.1021/acs.orglett.7b03801DOI Listing
February 2018

miR-324-3p promotes gastric cancer development by activating Smad4-mediated Wnt/beta-catenin signaling pathway.

J Gastroenterol 2018 Jun 4;53(6):725-739. Epub 2017 Nov 4.

Department of General Surgery, The First Affiliated Hospital of Nanjing Medical University, No. 300 Guangzhou road, Nanjing, Jiangsu, China.

Background: Emerging evidence suggested that miRNAs can function as oncogenes or tumor suppressors by regulating downstream target genes. miR-324-3p has been reported to function in several carcinomas, but its role in gastric cancer (GC) is still unknown. This study aims to explore the effects of miR-324-3p on the development of GC.

Methods: Expression of miR-324-3p was examined in GC cells and tissues by qRT-PCR. Effects of miR-324-3p on GC cells were evaluated by cell vitality assay, colony formation assay, cell migration assay, and flow cytometric assay. The dual luciferase assay was used to verify whether miR-324-3p could interact with the potential target genes. Western blot was used to assess the expression level of Smad4 and beta-catenin. Intracellular ATP level was also examined. The tumor xenografts were established using nude mice. A gastric organoid model was made from fresh stomach tissue.

Results: miR-324-3p was expressed at higher levels in the tumor tissues compared with adjacent normal tissues. Overexpression of miR-324-3p promoted cell growth, migration, and decreased apoptosis. miR-324-3p repressed the expression of Smad4, and loss of Smad4 activated the Wnt/beta-catenin signaling pathway. Overexpression of Smad4 rescued the effects of miR-324-3p on GC cells. The intracellular ATP level was upregulated with overexpression of miR-324-3p. miR-324-3p facilitated tumor cell colonization and growth in vivo and contributed to the growth of gastric organoids.

Conclusions: The results suggested that miR-324-3p promoted GC through activating the Smad4-mediated Wnt/beta-catenin signaling pathway. The miR-324-3p/Smad4/Wnt signaling axis may be a potential therapeutic target to prevent GC progression.
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http://dx.doi.org/10.1007/s00535-017-1408-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5971041PMC
June 2018

Palmitoylethanolamide (PEA): A promising biomarker for coronary dysfunction in MOB individuals.

Int J Cardiol 2017 09;242:26

Department of Cardiovascular Surgery, Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, China.

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http://dx.doi.org/10.1016/j.ijcard.2017.04.098DOI Listing
September 2017

Natriuretic peptide receptor A inhibition suppresses gastric cancer development through reactive oxygen species-mediated G2/M cell cycle arrest and cell death.

Free Radic Biol Med 2016 10 12;99:593-607. Epub 2016 Sep 12.

Department of General Surgery, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China; Collaborative Innovation Center For Cancer Personalized Medicine, Nanjing Medical University, Nanjing, China. Electronic address:

Natriuretic peptide receptor A (NPRA), the major receptor for atrial natriuretic peptide (ANP), has been implicated in tumorigenesis; however, the role of ANP-NPRA signaling in the development of gastric cancer remains unclear. Immunohistochemical analyses indicated that NPRA expression was positively associated with gastric tumor size and cancer stage. NPRA inhibition by shRNA induced G2/M cell cycle arrest, cell death, and autophagy in gastric cancer cells, due to accumulation of reactive oxygen species (ROS). Either genetic or pharmacologic inhibition of autophagy led to caspase-dependent cell death. Therefore, autophagy induced by NPRA silencing may represent a cytoprotective mechanism. ROS accumulation activated c-Jun N-terminal kinase (JNK) and AMP-activated protein kinase (AMPK). ROS-mediated activation of JNK inhibited cell proliferation by disturbing cell cycle and decreased cell viability. In addition, AMPK activation promoted autophagy in NPRA-downregulated cancer cells. Overall, our results indicate that the inhibition of NPRA suppresses gastric cancer development and targeting NPRA may represent a promising strategy for the treatment of gastric cancer.
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http://dx.doi.org/10.1016/j.freeradbiomed.2016.08.019DOI Listing
October 2016

Purification and characterization of cholecystokinin from the skin of salamander Tylototriton verrucosus.

Dongwuxue Yanjiu 2015 May;36(3):174-7

Key Laboratory of Animal Models and Human Disease Mechanisms of Chinese Academy of Sciences & Yunnan Province, Kunming Institute of Zoology, Kunming Yunnan 650223, China.

As a group of intestinal hormones and neurotransmitters, cholecystokinins (CCKs) regulate and affect pancreatic enzyme secretion, gastrointestinal motility, pain hypersensitivity, digestion and satiety, and generally contain a DYMGWMDFG sequence at the C-terminus. Many CCKs have been reported in mammals. However, only a few have been reported in amphibians, such as Hyla nigrovittata, Xenopus laevis, and Rana catesbeiana, with none reported in urodele amphibians like newts and salamanders. Here, a CCK called CCK-TV was identified and characterized from the skin of the salamander Tylototriton verrucosus. This CCK contained an amino acid sequence of DYMGWMDF-NH2 as seen in other CCKs. A cDNA encoding the CCK precursor containing 129 amino acid residues was cloned from the cDNA library of T. verrucosus skin. The CCK-TV had the potential to induce the contraction of smooth muscle strips isolated from porcine gallbladder, eliciting contraction at a concentration of 5.0 x 10⁻¹¹ mol/L and inducing maximal contraction at a concentration of 2.0 x 10⁻⁶ mol/L. The EC50 was 13.6 nmol/L. To the best of our knowledge, this is the first report to identify the presence of a CCK in an urodele amphibian.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4790693PMC
May 2015

Altered Fruit and Seed Development of Transgenic Rapeseed (Brassica napus) Over-Expressing MicroRNA394.

PLoS One 2015 15;10(5):e0125427. Epub 2015 May 15.

Department of Biochemistry and Molecular Biology, College of Life Science, Nanjing Agricultural University, Nanjing, China.

Fruit and seed development in plants is a complex biological process mainly involved in input and biosynthesis of many storage compounds such as proteins and oils. Although the basic biochemical pathways for production of the storage metabolites in plants are well characterized, their regulatory mechanisms are not fully understood. In this study, we functionally identified rapeseed (Brassica napus) miR394 with its target gene Brassica napus leaf curling responsiveness (BnLCR) to dissect a role of miR394 during the fruit and seed development. Transgenic rapeseed plants over-expressing miR394 under the control of the cauliflower mosaic virus 35S promoter were generated. miR394 over-expression plants exhibited a delayed flowering time and enlarged size of plants, leaf blade, pods and seed body, but developed seeds with higher contents of protein and glucosinolates (GLS) and lower levels of oil accumulation as compared to wild-type. Over-expression of miR394 altered the fatty acid (FA) composition by increasing several FA species such as C16:0 and C18:0 and unsaturated species of C20:1 and C22:1 but lowering C18:3. This change was accompanied by induction of genes coding for transcription factors of FA synthesis including leafy cotyledon1 (BnLEC1), BnLEC2, and FUSCA3 (FUS3). Because the phytohormone auxin plays a crucial role in fruit development and seed patterning, the DR5-GUS reporter was used for monitoring the auxin response in Arabidopsis siliques and demonstrated that the DR5 gene was strongly expressed. These results suggest that BnmiR394 is involved in rapeseed fruit and seed development.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0125427PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4433277PMC
March 2016

The F-box family genes as key elements in response to salt, heavy mental, and drought stresses in Medicago truncatula.

Funct Integr Genomics 2015 Jul 16;15(4):495-507. Epub 2015 Apr 16.

Department of Biochemistry and Molecular Biology, College of Life Science, Nanjing Agricultural University, Weigang No. 1, Nanjing, 210095, China.

F-box protein is a subunit of Skp1-Rbx1-Cul1-F-box protein (SCF) complex with typically conserved F-box motifs of approximately 40 amino acids and is one of the largest protein families in eukaryotes. F-box proteins play critical roles in selective and specific protein degradation through the 26S proteasome. In this study, we bioinformatically identified 972 putative F-box proteins from Medicago truncatula genome. Our analysis showed that in addition to the conserved motif, the F-box proteins have several other functional domains in their C-terminal regions (e.g., LRRs, Kelch, FBA, and PP2), some of which were found to be M. truncatula species-specific. By phylogenetic analysis of the F-box motifs, these proteins can be classified into three major families, and each family can be further grouped into more subgroups. Analysis of the genomic distribution of F-box genes on M. truncatula chromosomes revealed that the evolutional expansion of F-box genes in M. truncatula was probably due to localized gene duplications. To investigate the possible response of the F-box genes to abiotic stresses, both publicly available and customer-prepared microarrays were analyzed. Most of the F-box protein genes can be responding to salt and heavy metal stresses. Real-time PCR analysis confirmed that some of the F-box protein genes containing heat, drought, salicylic acid, and abscisic acid responsive cis-elements were able to respond to the abiotic stresses.
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http://dx.doi.org/10.1007/s10142-015-0438-zDOI Listing
July 2015

[Voltage-gated sodium channels are involved in regulating the biological functions of microglial cells].

Sheng Li Xue Bao 2015 Feb;67(1):41-7

College of Life Science, Liaoning Normal University, Lamprey Research Center, Dalian 116081,

Microglial cells are widely distributed in the brain and spinal cord, and usually act as resident immune cells which could provide continuous monitoring roles within the central nervous system. When the cells in the central nervous system are injured, microglial cells are activated and induce a series of biological effects. Recently, several voltage-gated sodium channel subtypes were found to be expressed on the surface of the microglial cells which are able to participate in the regulation of the activation, phagocytosis, secretion of multiple cytokines/chemokines, migration, invasion of microglial cells, and etc. In the present study, the latest progresses on the regulation of voltage-gated sodium channel isoforms on microglial cells were summarized and analyzed. In addition, the mechanism and future research of the relationship between voltage-gated sodium channels and microglial cells were also discussed.
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February 2015

Synthesis, crystal structure, and biological activities of two chiral mononuclear Mn((III)) complexes.

Chirality 2015 Feb 18;27(2):142-50. Epub 2014 Nov 18.

Department of Chemistry, Nankai University, Tianjin, People's Republic of China; Tianjin Key Laboratory of Metal and Molecule Based Material Chemistry, Tianjin, People's Republic of China; Key Laboratory of Advanced Energy Materials Chemistry (MOE), Tianjin, People's Republic of China.

Two new chiral mononuclear Mn((III)) complexes, [MnL((R)) Cl (C2 H5 OH)]•C2 H5 OH () and [MnL((S)) (CH3 OH)2 ]Cl•CH3 OH (), {H2 L = (R,R)-or (S,S)-N,N'-bis-(2-hydroxy-1-naphthalidehydene)-cyclohexanediamine} were synthesized and characterized by various physicochemical techniques. Bond valence sum (BVS) calculations and the Jahn-Teller effect indicate that the Mn centers are in a +3 oxidation state. The statuses of the two complexes in the solution were confirmed as a pair of enantiomers by electrospray ionization, mass spectrometry (ESI-MS) spectrum. The binding ability of the complexes with calf thymus CT-DNA was investigated by spectroscopic and viscosity measurements. Both of the complexes could interact with CT-DNA via an intercalative mode with the order of (R-enantiomer) > (S-enantiomer). Under the physiological conditions, the two compounds exhibit efficient DNA cleavage activities without any external agent, which also follows the order of R-enantiomer > S-enantiomer. Interestingly, the concentration-dependent DNA cleavage experiments indicate an optimal concentration of 17.5 μM. In addition, the interaction of the compounds with bovine serum albumin (BSA) was also investigated, which indicated that the complexes could quench the intrinsic fluorescence of BSA by a static quenching mechanism.
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http://dx.doi.org/10.1002/chir.22403DOI Listing
February 2015
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