1.Computational Identification of microRNAs and Their Targets
Zhiyun GUO ; Canquan MAO ; Lili XIONG
China Biotechnology 2008;28(10):118-123
The discovery of microRNAs (miRNAs) has introduced a new paradigm into gene regulatory systems. Since inception, computational methods have been an invaluable tool complementing experimental approaches, and many discoveries have been obtained through combination of experimental and computational approaches. The knowledge that has been accumulated about the principles of miRNAs and target recognition were reviewed. The currently available computational methodologies and software for prediction of miRNA and their target genes also have been discussed.
2.NF-κB signaling pathway is involved in retinoic acid inhibiting dendritic cell differentiation and antigen presentation
Guoping LIU ; Jie GUO ; Zhiyun WANG ; Dongchun LIANG
Chinese Journal of Microbiology and Immunology 2012;32(7):622-628
Objective To clarify whether the regulatory effect of all-trans-retinoic acid (atRA) on the antigen presentation function of dendritic cell(DC) is tightly associated with NF-κB signaling pathway.Also to clarify atRA mainly affected differentiated DC or influence the differentiation procedure from bone marrow cell to DC.Methods MOG35-55 immunized C57BL/6J mice received administration of atRA or not,splenic DC and CD4 cells isolated from immunized mice were subjected to in vitro cross-culture,treated with IL-12 and IL-23 respectively.Th1 and Th17 polarization of stimulated CD4 cells were determined by intracellular staining and FACS analysis,while the production of their corresponding cytokines,IFN-γ and IL-17,were measured by ELISA.Bone marrow cells were isolated from the femurs of the na(i)ve mice,treated with IL-4 and GM-CSF with or without synergistic RA treatment.MHC Ⅱ and CD11c molecules on the DC were assayed by immune staining and FACS analysis,their antigen presentation functions were decided by the proliferation and cytokine production of the Th1 effecter cells stimulated by antigen pulsed DC.To investigate the status of the NF-κB signaling pathway,the amount of phospho-Ser536 NF-κB p65 in the whole DC lysate and total NF-κB p65 in the nucleus were detected by Western blot.Finally,selective RA receptor antagonists were studied to figure out which receptor was majorly involved in the regulatory effect of RA on DC.Results Splenic DC from RA treated mice showed significantly decreased function of presenting antigen to stimulate CD4 cell polarization and cytokine production.Compared with untreated control,RA in vitro treated DC showed decreased expression of MHC Ⅱ and CD11c on its cell surface,which was accompanied with depressed function of stimulating Th1 cell proliferation and IFN-γ production.Further study revealed that RA mainly affect the differentiation procedure from BMC to DC,however,has no significant effect on differentiated DC as the aspects of MHC Ⅱ,CD11c expression,responder cell proliferation and cytokine production were evaluated.Decreased amount of phosphor-Ser536 NF-κB p65 in the whole cell lysate and total NF-κB p65 in the nucleus was investigated in RA treated DC,and decreased antigen presentation function of RA treated DC always came together with diminished activation of NF-κB signaling pathway.Finally it was demonstrated that RARβγ antagonist,but not RARα antagonist,could entirely block the RA effect on DC.Conclusion Retinoic acid inhibit the differentiation of DC as well as decrease its antigen presentation function,which might be resulted from the inactivation of NF-κB p65 signaling pathway and mediated by RARβγ.
3.Compatible Stability of Cefmenoxime for Injection with Ganciclovir for Injection
Jianxin WANG ; Zhiyun HUANG ; Junli GUO ; Jizhang YANG
China Pharmacy 2015;(29):4066-4068
OBJECTIVE:To study the compatible stability of Cefmenoxime for injection with Ganciclovir for injection in 0.9% Sodium chloride injection and 5% Glucose injection. METHODS:At room temperature,the appearance and pH of the mix-tures were observed after Cefmenoxime for injection was compatible with Ganciclovir for injection. HPLC method was adopted to determine the content of them. RESULTS:No significant change was noted for the mixture in appearance and pH value. The con-tent of ganciclovir was more than 98%,but that of cefmenoxime decreased to 75.33%. CONCLUSIONS:Cefmenoxime for injec-tion can not be mixed with Ganciclovir for injection in 0.9%Sodium chloride injection and 5%Glucose injection .
4.A PHARMACODYNAMIC STUDY ON ZHI TAN LING
Guoxi ZHANG ; Zhiyun LIU ; Jian GUO ; Lanfei FENG ; Xiaohong SHANG ; Jiangang LIU ; Pu GAO ; Wenquan ZHOU ;
Traditional Chinese Drug Research & Clinical Pharmacology 1993;0(04):-
This paper presents the results of pharmacological studies on Zhi Tan Ling (ZTL). The results showed that ZTL could decrease the ischemic damage of the brain tissue in case of acute cerebral ischemia and increase synthetic metabolism, promote restoration of Cerebral thrombosis, increase cerebral blood flow, improve hemorheologic indexes and meningeal microcirculation in experimental animals. It is indicated that ZTL is effective for ischemic apoplexy through various mechanisms.
5.Toll like receptors signaling pathways directly increase the expression of functional IL-17RA in neu-roglial cells
Shumin ZHOU ; Song CHEN ; Guoping LIU ; Jie GUO ; Zhiyun WANG ; Dongchun LIANG
Chinese Journal of Microbiology and Immunology 2014;(3):179-185
Objective To investigate whether toll like receptor ( TLR) signaling pathways can in-crease the expression of IL-17 R in neuralglial cells , and if they can whether the increased IL-17 R is func-tional.Methods Experimental autoimmune encephalomyelitis (EAE) was induced in B6 mice by immuni-zation with an emulsion of myelin oligodendrocyte glycoprotein 35-55 ( MOG35-55 ) in complete Freund's adju-vant (CFA).The expression of Il17ra and Il17rc in the brains and spinal cords of mice with EAE were de-tected by real-time PCR.Luxol fast blue ( LFB) staining was performed to the spinal cord sections to detect tissue demyelination.Immunohistological staining against IL-17RA and CD3 were undertook to visualize IL-17RA+and CD3 + cells.Same approaches were also applied to immunized Rag1 -/- mice to figure out whether T cells infiltration is necessary for increasing IL-17RA expression in the central nervous system ( CNS) .Then B6 mice were immunized with incomplete Freund′s adjuvant ( IFA) plus different TLRs ago-nists to measure the expression of Il17ra in the brains and spinal cords by qPCR .The purified astrocytes , microglia and oligodendrocytes isolated from neonatal mice brains were cultured in vitro for two weeks , and then treated with different TLRs agonists .The expression of Il17ra at mRNA and protein levels in the cells were determined by qPCR and Western blot respectively .The astrocytes were treated with IL-17A and LPS individually or in the combination to detect the level of CCL 2, CXCL8 and IP-10 in the supernatant by ELISA.Results B6 mice with induced EAE showed significantly increased Il17ra expressions in the brain and spinal cord , which was also detected in immunized Rag1 -/-mice.Although no spinal cord demyeliza-tion and CD3 cells infiltration were detected in Rag1 -/-mice, significantly increased number of IL-17RA positive cells could still be visualized .In vivo TLRs agonist participated immunization and in vitro treatment of purified neuroglial cells demonstrated that TLRs agonists could directly evoke IL -17RA expression in the CNS or cultured astrocytes , microglia and oligodendrocytes with high efficiency .Both IL-17 A and LPS could stimulate astrocytes to secrete CCL2, CXCL8 and IP-10, however, a combined use of IL-17A and LPS fur-ther augmented the production of these chemokines to a large extend .Conclusion Taken together , we con-cluded that TLRs agonists could directly stimulate neuroglial cells to express IL -17RA which functionally re-spond to IL-17A by secreting chemokines .
6.Screening and validation of markers related to diagnosis and prognosis assessment of gastric cancer based on miRNA-mRNA network and their potential molecular mechanisms
Chinese Journal of Cancer Biotherapy 2022;29(12):1094-1100
[摘 要] 目的:通过生物信息学方法探索并实验验证胃癌相关标志物miR-1-3p对胃癌细胞增殖的作用及其分子机制。方法:收集TCGA数据库中胃癌(n=375)及癌旁组织(n=45)的转录组数据,构建胃癌特异性mRNA-miRNA网络,筛选潜在的miRNA类标志物,利用TargetScan预测标志物的下游靶基因且分析它们的功能。选取人正常胃上皮细胞GES-1及胃癌细胞AGS、MKN45、NCI-N87,用qPCR法检测细胞中miR-1-3p和心肌蛋白(MYOCD)的表达,用lipofectamine 2000将miR-1-3p模拟物转染至胃癌细胞中,CCK-8法测定轨染后细胞的增殖能力,WB法测定MYOCD的表达量,双荧光素酶报告基因实验验证miR-1-3p与MYOCD之间的靶向结合关系。结果:通过数据库数据分析得到差异表达的259个miRNA和7 545个mRNA,构建胃癌特异性mRNA-miRNA调节网络,分析网络中脆弱结构后确定miR-1-3p为潜在的胃癌标志物,ROC曲线和Kaplan-Meier分析显示其对胃癌的诊断和预后评估有重要意义。细胞实验显示miR-1-3p在胃癌细胞中呈低表达(P<0.05),过表达miR-1-3p可抑制胃癌细胞AGS和MKN-45的增殖能力(P<0.05或P<0.01),且可抑制MYOCD的表达(P<0.01)。TargetScan数据库预测到MYOCD的3'UTR区域中有两个与miR-1-3p结合的位点,双荧光素酶报告基因实验证实miR-1-3p与MYOCD靶向结合且负调控MYOCD的表达(P<0.01)。结论: miR-1-3p可能是胃癌诊断和预后相关潜在的标志物,且miR-1-3p可能是通过靶向MYOCD来影响胃癌细胞的增殖。
7.Development and application of bioelectric measurement system for vivo bone puncture.
Zhiyun WANG ; Qingkai DENG ; Qingshui YIN ; Jingsong GUO ; Jianbo WANG ; Shuofeng ZHAO
Chinese Journal of Medical Instrumentation 2010;34(3):164-166
Procedure of a bioelectric signal collection system for vivo critter is introduced in this paper. It is easy to measure the bioimpedance in the tip of appliance, when puncture into the tissue, especially puncture into the bone tissue. We can get a judgment on the position of appliance, thereby achieve assistance on the clinic operation.
Biopsy, Needle
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instrumentation
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Bone and Bones
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physiology
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Electrophysiological Phenomena
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Equipment Design
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Specimen Handling
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instrumentation
8. Analysis of blood flow energy characteristics of pulsatile and non-pulsatile flow during extracorporeal circulation
Zhen GUO ; Xin LI ; Lingfeng XU ; Xin CHANG ; Jian LI ; Zhiyun XU
Chinese Journal of Surgery 2018;56(9):701-705
Objective:
To analyze the magnitude of blood flow energy and characteristics of frequency domain between pulsatile flow and nonpulsatile flow during cardiopulmonary bypass and physiological flow.
Methods:
From January 2017 to December 2017, 60 cases of patients with mitral valve disease scheduled for mitral valve replacement or repair at Department of Cardiasurgery, Shanghai Chest Hospital, Shanghai Jiaotong University were randomly divided into 2 groups: pulsatile perfusion (PP) and non-pulsatile perfusion (NP). The magnitude of blood flow energy during pulsatile and non-pulsatile was calculated using energy equivalent pressure (EEP) and surplus hemodynamic energy (SHE) while fast Fourier transformation (FFT) was used to perform power spectral density analysis to identify the frequency domain characteristics between artificial and physiological flow (prior to CPB). The data was analyzed by analysis of variance or
9.The value of myeloid-derived suppressor cell and T lymphocyte subsets in the diagnosis and treatment of active pulmonary tuberculosis
Fei CHEN ; Jing GUO ; Zhiyun CAO ; Xianshu FEI
Chinese Journal of Postgraduates of Medicine 2022;45(3):207-210
Objective:To investigate the levels and correlation between myeloid-derived suppressor cell (MDSC) and T lymphocyte subsets in peripheral blood of patients with active pulmonary tuberculosis.Methods:A total of 38 patients with active pulmonary tuberculosis in Nanjing Second Hospital from February 2019 to June 2020 were selected as the tuberculosis group, and 23 healthy outpatient physical examination patients were selected as the healthy control group during the same period. The levels of MDSC, clinically related indicators, inflammatory cytokines and lymphocyte subsets were compared between each group, and the correlation between MDSC and lymphocyte subsets was analyzed. Meanwhile, the levels of MDSC and lymphocyte subsets before and after treatment were compared.Results:The MDSC and CRP in tuberculosis group were higher than those in healthy group: (16.41 ± 2.13)% vs. (1.82 ± 0.54)%, (25.42 ± 10.56) mg/L vs. (5.82 ± 1.39) mg/L ( P<0.05). Serum inflammatory cytokines tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-1β, IL-10 and interferon (IFN)-γ in tuberculosis group were significantly higher than those in healthy control group ( P<0.05). T lymphocyte subsets CD 3+ T cell, CD 4+ T cell, CD 8+ T cell and CD 16/56+ nature killer (NK) cell in tuberculosis group were significantly lower than those in healthy control group ( P<0.05), while the number of CD 19+ B cell was not statistically significant ( P>0.05). Correlation analysis showed that MDSC was negatively correlated with T lymphocyte subsets CD 3+ T cell ( r = -0.73, P<0.001), CD 4+ T cell ( r = -0.68, P<0.001) and CD 8+ T cell ( r = -0.53, P = 0.001), but had no significant correlation with CD 16/56+ NK cell ( r = -0.10, P = 0.561). CD 3+ T cell, CD 4+ T cell, CD 8+ T cell and CD 16/56+ NK cell were significantly different in peripheral blood MDSC before and after treatment ( P<0.05). Conclusions:MDSC, CD 3+ T cell, CD 4+ T cell, CD 8+ T cell and CD 16/56+ NK cell have a guiding role in the diagnosis and evaluation of the curative effect of active pulmonary tuberculosis, with high value in clinical application.
10.Identification and characteristic analysis of enhancer-miRNA regulatory pairs in hepatic carcinoma
TANG Fei ; WANG Wenzhu ; LANG Mei ; GUO Zhiyun
Chinese Journal of Cancer Biotherapy 2020;27(7):781-786
[Abstract] Objective: To explore the regulatory relationship between enhancer and miRNA and the characteristics of the enhancers that regulate miRNA in hepatic carcinoma and normal hepatic tissues, and to screen the differentially expressed miRNAs regulated by enhancers as well as their association with the treatment targets in liver cancer. Methods: Based on the TCGA and FANTOM5 databases, Co-expression and 3D genomic analysis were performed on 417 samples of enhancers and miRNAs in liver cancer and normal liver tissues. The difference in signal value of the enhancer that regulates miRNA was analyzed by ChIP-seq data of histone modification and transcription factor in liver cancer and normal liver tissues in ENCODE database. The differentially expressed miRNAs regulated by enhancers were screened out, and the correlation analysis was performed on the patient's survival and treatment targets. Results: 93 and 40 pairs of enhancer-miRNA were identified in liver cancer and normal liver tissues, respectively. ChIP-seqdata comparison analysis found that the signal of H3K27ac, H3K4me1 and sH3K4me3 histone modification in the region of enhancers
regulating miRNA was significantly higher than that in the region of enhancers not regulating miRNA (|rho|>0.3, P<0.05). Moreover, the enrichment of multiple transcription factors in liver cancer-related enhancers was significantly lower than that in normal liver tissue-realted enhancers (|rho|>0.3, P<0.05). Differential expression analysis of enhancer-regulated miRNAs identified 6 miRNAs related to the survival of liver cancer patients(hsa-miR-4664, hsa-miR-5003,hsa-miR-1915,hsa-miR-3619,hsa-miR-4745, hsa-miR-6728),and found that these miRNAs were significantly associated with 87 genes for targeted therapy and 8 tumor immune checkpoint genes (|rho|>0.1, FDR<0.05). Conclusion: The enhancer-miRNA regulatory pairs and the characteristics of the enhancer that regulates miRNA were successfully identified in liver cancer. The miRNAs regulated by enhancers and related to the therapeutic targets and survival of patients with liver cancer were also screened out. It provides a valuable preliminary basis for future in-depth basic and clinical research in hepatic carcinoma.