1.Gene expression of astrocytes after ischemic stroke
International Journal of Cerebrovascular Diseases 2015;23(11):877-880
Astrocytes play many important roles in the nervous system.In recent years,researchers are paying attention to the roles of astrocytes in the process of stroke.This article mainly reviews the biological functions of astrocytes and the gene expression changes of astrocytes after ischemic stroke,thereby providing new potential therapeutic targets for the treatment of ischemic stroke.
2.Academic thinking of LOU Duo-feng professor in treating rheumatism
Huiqin GUO ; Pei LI ; Yanhong LOU ; Yuqian LOU
China Journal of Traditional Chinese Medicine and Pharmacy 2005;0(06):-
The article summed up academic thinking of LOU Duo-feng professor in treating rheumatism.Professor LOU thought that deficiency,pathogen and stasis were the fundamental pathogenesis of rheumatism,and suggested that we should handle deficiency,pathogen and stagnation as a whole in treating rheumatism.There were four treatment principles should be obey in treating rheumatism:firstly,strengthening the genuine-qi to eliminate the evil-qi,simultaneous treatment of principal and subordinate symptoms;Secondly,application of diffusing and obstruction-removing,choosing different medicine according to different location of disease;Thirdly,treatment in accordance with three types of disease causes,treating combing similarities and differences;Lastly,changes of prescriptions and principals according to different courses of disease.In this article,the author list Tongbitang,the basic prescription of Professor LOU in treating rheumatism,and his clinical experience in treating deficiency heat type of obstinate arthralgia.
3.Prokaryotic expression of Streptococcus pneumoniae phpP gene and evaluation of PP2C type phosphatase activity of the expressed recombinant protein
Yanhong SUN ; Shaoni ZHANG ; Yongliang LOU ; Jie YAN ; Aihua SUN
Chinese Journal of Microbiology and Immunology 2014;(11):844-848
Objective To construct a prokaryotic expression system for expressing the phosphatase-encoding gene phpP in StkP/PhpP signaling couple in Streptococcus pneumonia ( S.pneumoniae) strains, and to further understand the phosphatase activity of the recombinant protein rPhpP.Methods The entire phpP gene of S.pneumoniae strain ATCC6306 was amplified by PCR.The PCR products were sequenced.A prokaryotic ex-pression system for expressing the phpP gene was constructed by the genetic engineering technique.The ex-pressed protein rPhpP and the solubility of rPhpP were assessed by SDS-PAGE and gel image analyzer.Ni-NTA affinity chromatography was performed to purify rPhpP.The changes of phpP gene transcription after the treat-ment with sublethal dosages of penicillin and cefotaxime were determined by real-time fluorescent quantitative RT-PCR.The functional domain in the sequence of the phpP gene and its type was analyzed by bioinformatic softwares.The activity of rPhpP in hydrolyzing the substrate of PP2C phosphotase was measured with Serine/Threonine Phosphotase Assay Kit.The enzyme kinetic parameters of rPhpP were calculated.Results The se-quence of the cloned phpP gene was identical with that reported in GenBank.The rPhpP in soluble form was ex-pressed in the constructed prokaryotic expression system.An increased expression of phpP gene at mRNA level was induced by sublethal dosage of penicillin or cefotaxime.The domain of PP2Cc type phosphatase was detec-ted in the sequence of phpP gene.The purified rPhpP protein could hydrolyze phosphopeptides [ RRA ( pT) VA], a substrate of PP2C type phosphatase, in a dose-dependent manner with Km and Kcat values of 277.35μmol/L and 0.71 S-1 ,respectively.Conclusion The protein encoded by phpP gene of S.pneumoniae was a PP2C type phosphatase.The expression of phpP gene could be enhanced by sublethal dosage of penicillin or ce-fotaxime.
4.Correlation between the ciaH gene of Streptococcus pneumoniae and the resistance toβ-lactam antibi-otics
Shaoni ZHANG ; Yanhong SUN ; Yongliang LOU ; Jie YAN ; Aihua SUN
Chinese Journal of Microbiology and Immunology 2015;(9):666-671
Objective To construct a mutant strain of Streptococcus pneumoniae ( S.pneumoniae) with ciaH gene-knockout (ΔciaH) and to analyze the correlation between the ciaH gene and the bacterial re-sistance against β-lactam antibiotics.Methods The ciaH gene segament of S.pneumoniae strain ATCC6306 was amplified by PCR.The PCR product was sequenced after T-A cloning.A suicide plasmid pEVP3ciaH was constructed for the deletion of ciaH gene and then transformed into the ATCC 6306 strain by using the CaCl2 method .The mutant strain of S.pneumonia strain ATCC6306 with ciaH gene-knockout (ΔciaH) was genera-ted through homologous recombination , insertion inactivation and amphemycin screening , which was further identified by PCR , sequencing analysis and laser confocal microscopy .Double agar dilution method was used to detect the minimal inhibitory concentrations ( MICs ) of penicillin G ( PCN ) and cefotaxime ( CTX ) against theΔciaH mutant strain and the wild type strain .The differences between the MICs were further ana-lyzed.The changes of ciaH gene expression at mRNA level after treatment with 1/4 MIC of PCN or CTX were detected by real-time fluorescent quantitative RT-PCR ( qRT-PCR ) .Results The ciaH gene in the genomic DNA of the generated ΔciaH mutant strain was inactivated by insertion as indicated by PCR and se-quencing analysis .Results of the immunofluorescence assay showed that the ΔciaH mutant strain did not ex-press the CiaH protein .The MICs of PCN and CTX against the ΔciaH mutant strain were 32 μg/ml and 64μg/ml, respectively, which were significantly higher than that of the wild type strain (0.06 μg/ml and 1μg/ml) (P<0.01).The expression of ciaH gene at mRNA level was significantly elevated after treatment S.pneumoniae ATCC6306 strain with 1/4 MIC PCN or CTX (P<0.01).Conclusion The CiaH protein in the CiaH/CiaR two-component signaling system is involved in the resistance of S.pneumoniae against β-lac-tam antibiotics.
5.Effects of Astragalus membranaceus and Potentilla discolor mixture on insulin resistance and its related mRNA expressions in KKAy mice with type 2 diabetes.
Dongmei ZHANG ; Lixia LOU ; Aiming WU ; Xiying Lü ; Zhaojuan HU ; Yanhong ZHANG ; Hongfang LIU
Journal of Integrative Medicine 2012;10(7):821-6
To investigate the effects of Astragalus membranaceus and Potentilla discolor mixture (APM) on insulin resistance (IR) and mRNA expressions of IR-related genes, including phosphatidylinositol 3-kinase (PI3-K), phosphoenolpyruvate carboxykinase (PEPCK) and peroxisome proliferator-activated receptor γ coactivator 1 (PGC1) in KKAy mice with early type 2 diabetes and to explore the gene regulation mechanisms of AMP.
6.Effect of rhynchophylline on the mRNA expression of excitatory amino acid transporter 2 and NMDA receptor 2B after oxygen-glucose deprivation in cultured rat hippocampal astrocytes
Yanhong CHEN ; Xiaoliang LOU ; Zhengcheng ZHOU ; Xueyong HU ; Qiujiang XI ; Hongwu YANG ; Lihua XIAO ; Xinhong LI ; Xiaoping LI ; Hua FAN
International Journal of Cerebrovascular Diseases 2018;26(9):684-688
Objective To investigate the effect of rhynchophylline on mRNA expression of excitatory amino acid transporter 2 (EAAT2 ) and N-methyl-D-aspartic acid receptor 2B (NR2B) after astrocyte oxygen-glucose deprivation. Methods The subcultured third generation astrocytes from the hippocampus were inoculated into 6-well plates, and they were divided into blank control group, hypoxia-ischemia group,low-dose rhynchophylline group (0. 02 mg/ml) and high-dose rhynchophylline group (0. 2 mg/ml) after the cells were attached to the wall and grew out protrusion. The total RNAs in each group were extracted.Real-time fluorescence quantitative polymerase chain reaction was used to detect the expression levels of EAAT2 and NR2B mRNA in astrocytes of each group. Results Compared with the blank control group, the expression levels of NR2B and EAAT2 mRNA in astrocytes of the ischemia-hypoxia group were significantly higher (all P < 0. 05 ). The expression levels of NR2B and EAAT2 mRNA in the low-dose rhynchophylline group were lower than those in ischemia-hypoxia group, but there was no significant difference. The expression levels of NR2B and EAAT2 mRNA in the high-dose rhynchophylline group were significantly lower than the ischemia-hypoxia group and the low-dose rhynchophylline group (all P < 0. 05).Conclusion The expression of EAAT2 and NR2B mRNA in astrocytes of hippocampus cultured in vitro was significantly increased after ischemia and hypoxia, and rhynchophylline intervention could significantly reduce its expression in a concentration dependent manner.
7.Establishment and evaluation of anesthesia nurses training system for postoperative delirium assessment
Yuxiang SONG ; Menglin CHI ; Yining SONG ; Min HOU ; Wei WANG ; Hao LI ; Jingsheng LOU ; Yanhong LIU ; Jiangbei CAO ; Weidong MI
Chinese Journal of Anesthesiology 2021;41(4):401-405
Objective:To establish training system for postoperative delirium (POD) assessment and evaluate the efficacy of training for anesthesia nurses.Methods:Sixteen nurse anesthetists of both sexes in our hospital were selected and received the systemic training for POD assessment.The training system included questionnaire survey, theoretical teaching, simulated visit, clinical observation, independent evaluation, centralized question-answering, evaluation of efficacy and random inspection.The level of POD knowledge tests were performed before the training and at the end of the fourth week of independent evaluation, respectively.At week 1 and 4 of independent evaluation, the diagnostic rate of POD and sensitivity and specificity of the assessment were calculated, and Kappa consistency analysis was used to assess the consistency between anesthesia nurses and training group in diagnosis of POD.In the first week of the third month after the end of training, the evaluation results were randomly inspected, the POD diagnosis rate was calculated between the anesthesia nurses and the training group, and the consistency analysis was conducted.Results:Compared with the scores of POD knowledge questionnaire and sensitivity of the assessment of the anesthesia nurses in the first week of training, the scores were significantly increased ( P<0.05), and no significant change was found in the POD diagnosis rate in the fourth week of training ( P>0.05). Compared with the training group, the diagnosis rate of POD of anesthesia nurses was significantly decreased in the first week of training ( P<0.05), and no significant change was found at the fourth week of training ( P>0.05). In the first and fourth weeks of training, the Kappa value of anesthesia nurses and the training group was 0.676 and 0.954 ( P<0.001), respectively.In the first week of the third month after the end of training, the Kappa value between anesthesia nurses and the training group in diagnosis of POD was 0.862 ( P<0.05). Conclusion:The training system of POD assessment has been successfully established, and the standardized anesthesia nurses training of POD has been achieved with good results.
8.Network pharmacology analysis and molecular docking verification of the mechanism of Zhenqi Fuzheng Capsule in the adjuvant treatment treatment of AIDS
Shengxing CAI ; Kaining WANG ; Yifang LOU ; Xiaonan HU ; Yanhong WANG ; Pei ZHOU ; Hao GU ; Xiaoping ZHANG ; Jian WANG ; Guojian GAO ; Yufeng ZHAO
International Journal of Traditional Chinese Medicine 2023;45(6):736-742
Objective:To analyze the molecular mechanism of Zhenqi Fuzheng Capsules in the adjuvant treatment of AIDS by network pharmacology method and molecular docking technology.Methods:The active components and targets of Zhenqi Fuzheng Capsules were obtained through TCMSP, and the AIDS-related targets were obtained through GeneCards, OMIM and DrugBank databases. The intersection target PPI network was constructed through STRING 11.5 database, and Cytoscape 3.9.1 software was used for network topology analysis; Metascape database was used for GO function and KEGG pathway enrichment analysis of core targets; Cytoscape 3.9.1 was used to construct Zhenqi Fuzheng Capsules component-target-pathway network; Autodock Tools software was used to carry out molecular docking of core targets and active components.Results:Totally 31 active components and 180 targets of Zhenqi Fuzheng Capsules were screened out. TNF, IL6, AKT1, IL1B, TP53, VEGFA, RELA, EGFR and CASP3 were identified as the core targets. GO functional enrichment analysis obtained 1 436 biological processes, 53 cellular components, and 117 molecular functions. KEGG pathway enrichment analysis obtained 167 pathways, which were related to pathways in cancer, AGE-RAGE signaling pathway in diabetic complications, and IL-17 signaling pathway was closely related. Molecular docking results showed that core targets such as AKT1 and TNF had good binding activity to quercetin, kaempferol, and luteolin.Conclusion:The main active components of Zhenqi Fuzheng Capsules in the adjuvant treatment of AIDS are quercetin, kaempferol and luteolin, which may treat AIDS through the IL-17 signaling pathway.