1.Aminoglycosides Modifying Enzyme Genes and intⅠ gene in Stenotrophomonas maltoplilia
Xiaowei DENG ; Danmei ZHANG ; Changting LIU ; Hongli ZHOU ; Ailing LI
Chinese Journal of Nosocomiology 2009;0(17):-
OBJECTIVE To study the aminoglycosides modifying enzyme genes and intⅠ gene in Stenotrophomonas maltophilia in Chinese Armed Police Forces General Hospital.METHODS The samples of 27 multi-resistant S.maltophilia were collected from inpatiens from Jan 2006 to Oct 2007 in this Hospital.The sensitivity of the isolates to 14 antibacterial agents was determined using a broth induction method.The aminoglycosides modifying enzyme genes and intⅠ 1 gene were detected by PCR.RESULTS The multi-drug resistance of S.maltophilia was a serious problem.In 27 strains of S.maltophilia,the positive ant(2″)-Ⅰ were in 5 strains(18.5%),aac(3)-Ⅱ in 3 strains(11.1%)and aac(6')-Ⅱ in 1 strain(3.7%).The positive intⅠ gene was found in 11 strains(29.6%).CONCLUSIONS Multi-resistant S.maltophilia resistant to aminoglycosides mainly due to the presence of aminoglycoside modifying enzymes ant(2″)-Ⅰ,aac(3)-Ⅱ and aac(6')-Ⅱ.The aminoglycoside modifying enzymes ant(3″)-Ⅰ and aac(6)-ⅠZ were not detected carrying IntⅠ would be the reason of S.maltophilia resistant to aminoglycosides.
2.N-n-butyl haloperidol iodide protectsH9c2 cardiac myocytes against hypoxia/reoxygenationinjury through mitochondria-dependent apoptotic pathway
Bin WANG ; Danmei HUANG ; Yuanhang WANG ; Qiaoling ZHOU ; Hong LIN ; Yanmei ZHANG ; Ganggang SHI ; Fuchun ZHENG
Chinese Pharmacological Bulletin 2017;33(6):819-823
Aim To investigate the effect of N-n-butyl haloperidol iodide(F2) on mitochondria-dependent apoptotic pathway of H9c2 cardiac myocytes during hypoxia/reoxygenation(H/R) injury.Methods The H/R models of H9c2 cardiac myocytes were established.The H9c2 cardiac myocytes were randomly divided into five groups: control group(C group), hypoxia/reoxygenation group(H/R group), F2 low concentration group(L), F2 medium concentration group(M), F2 high concentration group(H).Apoptotic rate was evaluated by flow cytometry(FCM).The levels of Cyto C, Bcl-2, Bax were observed by Western blot.Caspase-3 activity was measured with colorimetry.Results Compared with H/R group, F2 low, medium and high concentrations group could significantly decrease apoptosis rate and increase the ratio of Bcl-2 to Bax proteins and inhibit the release of Cyto C into the cytosolic fraction, and decrease caspase-3 activity.Conclusion F2 can protect H9c2 cardiac myocytes against H/R-induced injury through interfering in mitochondria-dependent pathway.
3.Understanding of the relations of metabolic syndromes and abnormal Bianzheng based on the physiological characteristics of the liver and spleen in children
Danmei CUI ; Xiaoke ZHANG ; Qiang ZHOU ; Yao CHEN
International Journal of Traditional Chinese Medicine 2022;44(3):241-245
Bianzheng was a special gasification law, closely related to metabolism Physiologically. The children's Bianzheng was the internal basis for sound physical and mental development and the dynamic balance of the five internal organs. As to the pathology, children's Bianzheng disorders had an adverse effect on the body. The ancient books found that children's Bianzheng disorders were closely related to the dysfunction of liver and spleen. It was manifested as malnutrition, developmental disorders, and even metabolic syndrome in adulthood. The principle of soothing the liver, relieving depression and strengthening the spleen were proposed to treat the metabolic syndrome, in order to enrich the Traditional Chinese Medicine (TCM) theoretical understanding of the metabolic syndrome. The prevention and treatment ideas provide theoretical reference.
4.Neuroepithelial tumor with EWSR1 translocation of central nervous system:a clinicopathological analysis of six cases
Danmei ZHOU ; Xueling QI ; Shujing ZHENG ; Xingfu WANG ; Yin WANG ; Feng TANG ; Ji XIONG ; Zunguo DU
Chinese Journal of Clinical and Experimental Pathology 2024;40(5):503-508
Purpose To investigate the clinical,imaging,pathological,and genetic characteristics of neuroepithelial tumors with EWSR1 translocation.Methods The clinicopatho-logical data of 6 patients with EWSR1 translocation in neuroepi-thelial tumors were collected,routine HE and immunohistochem-ical staining were performed,the information of high-throughput sequencing was summarized,and the relevant literature was re-viewed.Results The median age of the 6 patients was 11.5 years(ranging from 1.9 to 17 years),including 1 male and 5 females.The tumors located in temporal lobe,frontal lobe,pari-etal lobe,suprasellar region,or lateral ventricle.The clinical manifestations mainly started with seizures.Brain MRI showed abnormal signal focus in the cerebral hemisphere near the cortex in 4 cases,and ventricle/periventricular regions in 2 cases,with an almost clear boundary in 5 cases.Microscopically,the histo-logical changes were diverse,including low-grade gliomas/gli-oneuronal tumors in 3 cases,high-grade gliomas in 2 cases,and glioneuronal tumor with high-grade feature in 1 case.Immuno-histochemically,tumor cells expressed GFAP,S-100,Syn,and Olig2 partially.2 cases exhibited slightly positive of NeuN and 1 case exhibited little dot-like staining of EMA.Next generation sequencing revealed EWSR1 rearrangement in all 6 cases,with chaperone genes including PATZ1 in 5 cases,and PLAGL1 in 1 case.3 cases were treated with chemotherapy after surgery,and no recurrence or progression was found during follow-up.Con-clusion The neuroepithelial tumors with the fusion of EWSR1 and non-ETS commonly occur in the cerebral hemisphere of teenagers and children.Most of the boundaries lesion are still clear,the histomorphological spectrum is diverse,and the bio-logical behavior is presented as a low to moderate malignancy,which provides the possibility for expanding the molecular classi-fication of CNS neuroepithelial tumor.
5.3,5-Dihydroxy-4-methoxybenzyl alcohol attenuates hypoxic/reoxygenation injury of human umbilical vein endothelial cells
Zongmian YANG ; Mengjun ZHOU ; Yuling ZHANG ; Danmei HUANG ; Yang ZHANG ; WONG NAI-KEI ; Yanmei ZHANG
Chinese Journal of Pathophysiology 2024;40(11):2067-2073
AIM:To explore the effect of polyphenolic compound 3,5-dihydroxy-4-methoxybenzyl alcohol(DHMBA)on hypoxia/reoxygenation(H/R)injury of human umbilical vein endothelial cells(EA.hy926 cells)and its po-tential mechanisms.METHODS:To construct an H/R model,the EA.hy926 cells were cultured in an acidic hypoxia buffer while in an anaerobic workstation.The cells were divided into control,H/R,H/R+different doses of DHMBA,H/R+edaravone(antioxidant)and H/R+reactive oxygen species(ROS)inducer oligomycin A+DHMBA groups.Cell viability was measured by CCK-8 assay,and tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)levels in cells were mea-sured by ELISA.Phosphorylation of endothelial nitric oxide synthase(eNOS)and nuclear factor-κB(NF-κB)p65 were measured by Western blot.Intracellular NO levels were determined by laser confocal microscopy.Glutathione(GSH)/glu-tathione disulfide(GSSG)oxidation balance was determined by the dinitrobenzoic acid chromogenic method.Intracellular ROS levels were measured by flow cytometry.Lactate dehydrogenase(LDH)leakage was determined using nitro blue tet-razolium staining.Scratch assays were performed to assess cell migration.RESULTS:DHMBA exhibited no significant cytotoxicity between 125 and 1 000 μmol/L.In H/R-injured human umbilical vein endothelial cells,DHMBA improved cell survival,inhibited phosphorylation of NF-κB p65,reduced the content of TNF-α and IL-6,and increased phosphory-lation of eNOS and NO levels.DHMBA also suppressed ROS overload and restored the ratio between GSH and oxidized GSH,decreased in LDH leakage and increased cell migration in H/R-injured human umbilical vein endothelial cells.CONCLUSION:DHMBA can alleviate H/R-induced oxidative stress,inflammation,cellular damage,and dysfunction,which are associated with the ability of DHMBA to inhibit ROS production in human umbilical vein endothelial cells.
6.Screening and validation of key genes involved in necroptosis in atherosclerosis
Lijing YE ; Fuyang ZHOU ; Lexie BAI ; Chuangchuang GUO ; Shuaikai WU ; Yuzhi PAN ; Danmei WU ; Guojun ZHAO
Chinese Journal of Arteriosclerosis 2024;32(3):203-210
Aim The key genes for necroptosis in atherosclerosis were screened by bioinformatics methods and verified with the help of in vitro experiments to provide new strategies for the prevention and treatment of atherosclerosis from the perspective of necroptosis.Methods Genes related to atherosclerotic plaques were downloaded from GEO da-tabase,and genes related to necroptosis were downloaded from GeneCards database and intersected to obtain atherosclerotic necroptosis genes,and the mechanism of action and signalling pathways of the genes were further analysed by GO and KEGG enrichment analysis,and the protein-protein interaction(PPI)network was constructed and screened for key genes.Finally,macrophages were treated with oxidized low density lipoprotein(oa-LDL)at a final concentration of 100 mg/L,and the expression of key genes was detected by RT-PCR and Western blot.Results A total of 81 atherosclerotic nec-roptosis genes were obtained.GO and KEGG enrichment analyses revealed that they were mainly enriched in the positive regulation of endopeptidase activity,IκB kinase(IKK)/nuclear factor-KB(NF-κB)signalling,and autophagy signalling pathway.Five key genes including HSPA8,STAT3,HMOX1,SQSTM1 and FAS were obtained by using five computa-tional methods of Cytoscape software cytoHubba plug-in.Compared with the normal control group,the HMOX1 gene was highly expressed in THP-1 macrophages treated with ox-LDL(P<0.05),while the expression of the HSPA8,STAT3,SQSTM1 and FAS genes showed no significant changes(P>0.05);the HMOX1 and SQSTM1 genes were highly expressed in RAW264.7 macrophages treated with ox-LDL(P<0.05),while HSPA8,STAT3 and FAS genes showed no significant changes(P>0.05).The expression of HMOX1 protein in THP-1 macrophages was also increased.Conclusion HMOX1 may be the key gene of atherosclerotic necroptosis,and it is expected to become a new target for the prevention and treatment of atherosclerosis.
7.Deciphering chemical and metabolite profiling of Chang-Kang-Fang by UPLC-Q-TOF-MS/MS and its potential active components identification.
Fengge YANG ; Sihao ZHANG ; Danmei TIAN ; Guirong ZHOU ; Xiyang TANG ; Xinglong MIAO ; Yi HE ; Xinsheng YAO ; Jinshan TANG
Chinese Journal of Natural Medicines (English Ed.) 2023;21(6):459-480
Chang-Kang-Fang (CKF) formula, a Traditional Chinese Medicine (TCM) prescription, has been widely used for the treatment of irritable bowel syndrome (IBS). However, its potential material basis and underlying mechanism remain elusive. Therefore, this study employed an integrated approach that combined ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS) with network pharmacology to systematically characterize the phytochemical components and metabolites of CKF, as well as elucidating its underlying mechanism. Through this comprehensive analysis, a total of 150 components were identified or tentatively characterized within the CKF formula. Notably, six N-acetyldopamine oligomers from CicadaePeriostracum and eight resin glycosides from Cuscutae Semen were characterized in this formula for the first time. Meanwhile, 149 xenobiotics (58 prototypes and 91 metabolites) were detected in plasma, urine, feces, brain, and intestinal contents, and the in vivo metabolic pathways of resin glycosides were elaborated for the first time. Furthermore, network pharmacology and molecular docking analyses revealed that alkaloids, flavonoids, chromones, monoterpenes, N-acetyldopamine dimers, p-hydroxycinnamic acid, and Cus-3/isomer might be responsible for the beneficial effects of CKF in treating IBS, and CASP8, MARK14, PIK3C, PIK3R1, TLR4, and TNF may be its potential targets. These discoveries offer a comprehensive understanding of the potential material basis and clarify the underlying mechanism of the CKF formula in treating IBS, facilitating the broader application of CKF in the field of medicine.
Humans
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Tandem Mass Spectrometry/methods*
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Irritable Bowel Syndrome/drug therapy*
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Molecular Docking Simulation
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Drugs, Chinese Herbal/chemistry*
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Glycosides
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Chromatography, High Pressure Liquid/methods*