1.Effect of microRNA-7 knockdown on pathology of Enterotoxin-induced murine acute lung injury
Juanjuan ZHAO ; Hualin XU ; Mengmeng GUO ; Yijing TAO ; Ya ZHOU ; Chao CHEN ; Nalin QIN ; Jing ZHENG ; Dan TIAN ; Lin XU
Chinese Journal of Immunology 2016;32(9):1257-1261
Objective:To detect the effect of microRNA-7 ( miR-7 ) knockdown on pathology in murine acute lung injury ( ALI) model,and preliminarily explore its significance.Methods:Murine ALI model was performed by intraperitoneal injection of Li-popolysaccharide (LPS) (10 mg/kg) into miR-7KD mice and wild-type (wild type,WT) mice respectively.Then,the pathologic injury of lung tissue were observed by HE staining.And total cell count of bronchoalveolarlavage(BAL) was calculated.The relative expression of related cytokines in lung tissue was analyzed by Real-time PCR assay.Furthermore,the changes on proportion of innate immune cells (γδT cell and F4/80 macrophages cell) and adaptive immune cell ( CD4+T cell and CD8+T cell) were analyzed by FACS.Meanwhile, the expression of CD62L and CD69,as well as the absolute number,in CD4+T cell were also analyzed.Results: Compared with WT mice,pathological damage in lung tissues was significantly alleviated in miR-7KD mice.Real-time PCR analysis showed that the relative expression of IL-6 was obviously reduced (P<0.01),conversely,relative expression of IL-4 and TGF-βwere obviously increased (P<0.05).Furthermore,the total cell number in BAL also reduced significantly (P<0.05).Importantly,FACS analysis showed that the proportion and the absolute number of F4/80+Mφcells obviously reduced (P<0.05);however,the proportion of γδT cells increased (P<0.05).Moreover,the proportion and the absolute number of CD4+T cells and CD8+T cells were significantly reduced (P<0.05). Finally, the proportion and the absolute number of CD62L+in CD4+T cells were upregulated vigorously,contrastly,the proportion and the absolute number of CD69+in CD4+T cells were notably up-regulated (P<0.05).Conclusion:miR-7 defeciency could significantly ameliorate the pathology of murine ALI,suggesting that it may play an important regulatory role in the development of ALI.
3.Multi-locus sequence typing of multidrug-resistant of Acinetobacter baumannii from China and characterization of population structure of Acinetobacter baumannii.
Chao YANG ; Yan-feng YAN ; Gui-qin WANG ; Ya-fang TAN
Chinese Journal of Preventive Medicine 2011;45(8):727-731
OBJECTIVETo characterize the genetic background of multidrug-resistant Acinetobacter baumannii (A. baumannii) from China, and the population structure of this pathogen.
METHODSA previously reported MLST scheme was applied to a collection of 33 multidrug-resistant strains of A. baumannii from China, and the data of all the strains in the A. baumannii MLST database were downloaded for the population structure analysis. The sequence types and clonal complexes were identified, the presence or absence of recombination was analyzed for each MLST locus, and the values of I(A)(S), and recombination/mutation ratio were calculated for the whole strain collection. A phylogenetic tree was constructed using all the allelic profiles in the database.
RESULTSA total of six sequence types were identified from the 33 Chinese strains tested, and 29 of these strains belonged to the CC92 clonal complex. Three (gdhB, gpi, and rpoD) of the seven MLST loci (gltA, gyrB, recA, cpn60, gdhB, gpi, rpoD) had undergone recombination with statistical evidence. For all allele profiles in the MLST database, the I(A)(S) value was 0.155 and the recombination/mutation ratio was 6.083. Sequence types from each clonal complex were grouped closely in the phylogenetic tree, which gave an overview of the microevolution of this pathogen.
CONCLUSIONThe spread of multidrug-resistant A. baumannii in China was closely related to the CC92 clonal complex. A. baumannii had an 'epidemic' population structure, i.e., a superficially clonal structure with high levels of recombination, in which successful epidemic clones arise especially including worldwide dissemination of the CC92 clonal complex to cause a widespread occurrence of multidrug-resistant infections.
Acinetobacter baumannii ; classification ; genetics ; isolation & purification ; Bacterial Typing Techniques ; China ; Cluster Analysis ; DNA, Bacterial ; genetics ; Drug Resistance, Multiple, Bacterial ; Genetic Variation ; Genetics, Population ; Molecular Epidemiology ; Molecular Sequence Data ; Multilocus Sequence Typing ; Phylogeny
4.Cellular fatty acids as chemical markers for differentiation of Acinetobacter baumannii and Acinetobacter calcoaceticus.
Chao YANG ; Zhao Biao GUO ; Zong Min DU ; Hui Ying YANG ; Yu Jing BI ; Gui Qin WANG ; Ya Fang TAN
Biomedical and Environmental Sciences 2012;25(6):711-717
OBJECTIVEGas chromatography (GC) was used to investigate the cellular fatty acid (CFA) composition of 141 Acinetobacter baumannii and 32 A. calcoaceticus isolates from different locations in China and to find chemical markers to differentiate these two closely related bacteria.
METHODSWhole cell fatty acid methyl esters (FAMEs) were obtained by saponification, methylation, and extraction for GC analysis, followed by a standardized Microbial Identification System (MIS) analysis.
RESULTSAll A. baumannii and A. calcoaceticus strains contained some major fatty acids, namely, 18:1 ω9c, 16:0, Sum In Feature 3, 12:0, 17:1ω8c, 3-OH-12:0, 17:0, Sum In Feature 2, 2-OH-12:0, and 18:0 compounds. Although most of the total CFAs are similar between A. baumannii and A. calcoaceticus strains, the ratios of two pairs of CFAs, i.e., Sum In Feature 3/18:1 ω9c versus 16:0/18:1 ω9c and Sum In Feature 3/18:1 ω9c versus unknown 12.484/18:1 ω9c fatty acids, could differentiate these two closely related bacteria. A. baumannii could be easily classified into two subgroups by plotting some ratios such as Sum In Feature 3/16:0 versus 17:0 and Sum In Feature 3/2-OH-12:0 versus 17:0 fatty acids.
CONCLUSIONThe ratios of some CFAs could be used as chemical markers to distinguish A. baumannii from A. calcoaceticus.
Acinetobacter baumannii ; classification ; cytology ; metabolism ; Acinetobacter calcoaceticus ; classification ; cytology ; metabolism ; Biomarkers ; metabolism ; Fatty Acids ; metabolism ; Species Specificity
6.Establishment of a mouse hepatocellular carcinoma cell line producing mMIP-1α chemokines and the tomorigenicity of mMIP-1α transfected Hepa1-6
Lin-Hua QIN ; Qing YANG ; Li-Xin WEI ; Meng-Chao WU ; Yan LU ; Ya-Jun GUO
Academic Journal of Second Military Medical University 2001;22(5):418-421
Objective: To establish a mouse hepatocellular carcinoma cell line that can produce mMIP-1α and to evaluate the possibility of cancer gene therapy by mMIP-1α. Methods: mMIP-1α cDNA was cloned into retrovirus vector pBabe puro and pBabe puro-mMIP-1α was constructed, then pBabe puro-mMIP-1α was used to transfect packaging cells, anti-puromycin cells was proliferated, the supernatant was used to infect hepa1-6, the anti-puromycin clone (hepa1-6 mMIP-1α) and hepa1-6 were analysed for the expression of mMIP-1α mRNA and protein by RT-PCR and immunohistochemistry respectively. The growth curve of hepa1-6 and hepa1-6 mMIP-1α was drawn. The chemotaxis of mMIP-1α produced by hepa1-6 mMIP-1α to mouse spleen cells was observed on agarose gel. C57B/L mouse was inoculated with the tumor cell and the tumorigenicity was studied. Results: Recombinant retrovirus vector pBabe puro-mMIP-1α with mMIP-1α cDNA was constructed. Hepa1-6 did not produce mMIP-1α mRNA and protein, while hepa1-6 mMIP-1α could produce mMIP-1α mRNA and protein. The growth curve of hepa1-6 and hepa1-6 mMIP-1α showed no difference. The chemotaxis of mMIP-1α produced by hepa1-6 mMIP-1α to mouse spleen cells was observed. The tumorigenicity was reduced. Conclusion: A mouse hepatocellular carcinoma Hepa1-6 mMIP-1α is established and mMIP-1α can affect the tumorigenecity of hepa1-6.
7.Risk Factors, Clinical Features and Prognosis for Subtypes of Ischemic Stroke in a Chinese Population
Ya-Fu TAN ; Li-Xuan ZHAN ; Xiao-Hui CHEN ; Jian-Jun GUO ; Chao QIN ; En XU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2018;38(2):296-303
It is unanimously accepted that stroke is a highly heterogeneous disorder.Different subtypes of ischemic stroke may have different risk factors,clinical features,and prognoses.The aim of this study was to evaluate the risk factors,clinical characteristics,and prognoses of different subtypes of ischemic stroke defined by the Trial of ORG10172 in Acute Stroke Treatment (TOAST) criteria.We prospectively analyzed the data from 530 consecutive patients who were admitted to our hospital with acute ischemic stroke within 7 days of stroke onset during the study period.Standardized data assessment was used and the cause of ischemic stroke was classified according to the TOAST criteria.Patients were followed up till 30 and 90 days after stroke onset.It was found that large-artery atherosclerosis was the most frequent etiology of stroke (37.4%),and showed the highest male preponderance,the highest prevalence of previous transient ischemic attack,and the longest hospital stay among all subtypes.Small artery disease (36.4%) was associated with higher body mass index,higher plasma triglycerides,and lower plasma high-density lipoprotein cholesterol than cardioembolism.Cardioembolism (7.7%),which was particularly common in the elderly (i.e.,individuals aged 65 years and older),showed the highest female preponderance,the highest prevalence of atrial fibrillation,the earliest presentation to hospital after stroke onset,the most severe symptoms on admission,the maximum complications associated with an adverse outcome,and the highest rate of stroke recurrence and mortality.Our results suggest that ischemic stroke should be regarded as a highly heterogeneous disorder.Studies involving risk factors,clinical features,and prognoses of ischemic stroke should differentiate between etiologic stroke subtypes.
8.Identification of cattail pollen (puhuang), pine pollen (songhuafen) and its adulterants by ITS2 sequence.
Xiao-Xi MA ; Wei SUN ; Wei-Chao REN ; Li XIANG ; Bo ZHAO ; Ya-Qin ZHANG ; Ming SONG ; Ze-Jing MU ; Shi-Lin CHEN
China Journal of Chinese Materia Medica 2014;39(12):2189-2193
DNA barcoding method was conducted for the authentication of pollen materials due to difficulty of discriminating pollen materials bearing morphological similarity. In this study, a specific focus was to identify cattail pollen (Puhuang) and pine pollen (Songhuafen) samples from their adulterants which are frequently mixed-together. Regions of the internal transcribed spacer (ITS2) from 60 samples were sequenced, and new primers for cattail pollen were designed according to the sequence information. The results from the NJ trees showed that the species of pine pollen, Puhuang and their adulterants can be classified as obvious monophyly. Therefore, we propose to adapt DNA barcoding methodology to accurately distinguish cattail pollen, pine pollen and their adulterant materials. It is a great help for drug regulatory agency to supervise the quality of medicinal materials.
China
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DNA Barcoding, Taxonomic
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methods
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DNA, Plant
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genetics
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DNA, Ribosomal Spacer
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genetics
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Drug Contamination
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prevention & control
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Drugs, Chinese Herbal
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chemistry
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classification
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Molecular Sequence Data
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Phylogeny
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Pinus
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classification
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genetics
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Pollen
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classification
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genetics
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Quality Control
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Typhaceae
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classification
;
genetics
9.Study on preparation and thermosensitive release property of composite phospholipid liposomes containing total alkaloids from Strychnos nux-vomica.
Chao-Qin HE ; Meng-Ya HU ; Hui ZHANG ; Hao CHANG ; Jun CHEN ; Bao-Chang CAI
China Journal of Chinese Materia Medica 2013;38(9):1366-1370
To prepare composite phospholipid liposomes containing total alkaloids of Strychnos nux-vomica with hydrogenated soybean phosphatidylcholine (HSPC) and 1, 2-dipalmitoyl-sn-glycero-3-phosphacholine (DPPC), and compare with normal DPPC thermosensitive liposomes for thermosensitive release property. Total alkaloids were extracted from S. nux-vomica with the impregnation method and further purified. Liposomes containing total alkaloids, thermosensitive liposomes and conventional thermosensitive liposomes without thermosensitive release property were prepared by ammonium sulfate transmembrane gradients and stealth liposome technique. Their encapsulation efficiency (EE), grain size, zeta potential and drug release behavior were compared. Their EEs and zeta potentials were almost identical; but the grain sizes of composite phospholipid liposomes and thermosensitive liposomes were significantly smaller than conventional liposomes. After comparing release behaviors of the three liposomes at 37, 43 degrees C, we found that the release of composite phospholipid liposomes was significantly lower than that of thermosensitive liposomes at 37 degrees C, but higher than that of thermosensitive liposomes at 43 degrees C. Meanwhile, conventional liposomes, with a very high phase-transition temperature, showed only slight release behavior at both temperatures. The study results showed that composite phospholipid liposomes had a better thermosensitive release behavior when the dosage of lysophosphatidic was reduced by 2. 5 times.
Alkaloids
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chemistry
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Liposomes
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chemistry
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Phospholipids
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chemistry
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Strychnos nux-vomica
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chemistry
10.The relationship between TGF-?signal transduction pathway and pathogenesis of gastric carcinoma
Jian-Hong GUO ; Tao MA ; Yun-Peng ZHANG ; Wei-Qing DONG ; Tao FENG ; Ya-Tu GUO ; Xing-Yu LIANG ; Chao WANG ; Wei-Shan QIN ; Jian-Jin GUO
Cancer Research and Clinic 1997;0(03):-
Objective To study the relationship between TGF-?signaling pathway and pathogenesis of gastric carcinoma.Methods The expression of TGF-?RⅠ,TGF-?RⅡand Smad4 protein was deter- mined by immunohistochemistry in normal gastric mucosa(26 cases),intestinal metaplasia(22 cases),dysplasia (20 cases)and gastric carcinoma(43 cases).Results The positive expression rate of TGF-?RⅠ,TGF-?RⅡand Smad4 decreased following the malignant degree in gastric tissues(P