2.Comparison of Three Methods on Relatedness Analysis of Acinetobacter baumannii Strains from Hospital Infection
Xiaomin XU ; Lin CHEN ; Zuhuang MI ; Hui JIN ; Shun ZHANG
Chinese Journal of Nosocomiology 2006;0(06):-
OBJECTIVE To compare the three methods on relatedness analysis of Acinetobacter baumannii strains in hospital infection.METHODS Twenty-seven A.baumannii strains caused hospital infection were analyzed by pulsed field gel electrophoresis(PFGE),amplified fragment length polymorphism(AFLP) and multiple gene cluster analysis.RESULTS PFGE analysis showed 19 strains isolated from clinic were with identical clone;AFLP analysis showed 19 strains isolated from clinic were with identical clone;but multiple gene cluster analysis showed 3 clones including 11 strains carried with 8 positive genes(TEM,OXA-23group,ADC,aac(3)-Ⅰ,aac(6′)-Ⅰ,ant(3″)-Ⅰ,qacE△1-sul1 and intⅠ1),6 strains with 7 positive genes(TEM,ADC,aac(3)-Ⅰ,aac(6′)-Ⅰ,ant(3″)-Ⅰ,qacE△1-sul1 and intⅠ1) and 4 strains with 6 positive genes(TEM,ADC,aac(3)-Ⅰ,ant(3″)-Ⅰ,qacE△1-sul1 and intⅠ1).CONCLUSIONS The resolving ability of multiple gene cluster analysis is higher than that of PFGE and AFLP in relatedness analysis of A.baumannii strains from hospital infection.
3.Ecological distribution and diversity of medical Ferula species produced in Xinjiang.
Jun ZHU ; Xiao-Jin LI ; Li SUN ; Shun-xing GUO ; Juan CHEN
China Journal of Chinese Materia Medica 2015;40(2):356-361
To study the ecological distribution and diversity of endophytic fungi associated with Ferula of medicinal plants in Xinjiang. The endophytic fungi were isolated from roots, stems and leaves of Ferula by microbiology research methods and technology. The endophytic fungi were identified using ITS rDNA sequence analysis and morphology analysis. The composition, diversity and preference of endophytic fungal community were analyzed with Shannon-Wiener biodiversity index (H') and Sorensen coefficient (Cs). A total of 337 strains endophytic fungi were isolated and classified into 38 genera, Alternaria, Aureobasidium and Fusarium were the dominant genera. Among the 337 isolates, the endophytic fungi of F. sinkiangensis were the most, The Shannon-Wiener biodiversity index (H') associated with roots of F. fukanensis was the highest, reached 1.85. The highest Sorensen coefficient ( Cs) was between leaf of F. sinkiangensis and leaf of F. ovina, reached 0.75. From the result, endophytic fungi were widely distributed in six Ferula, there are some notable differences between distribution and composition of the endophytic fungi isolated from different issues and different species of Ferula, show a certain degree of species and tissue preference. The results obtained in this study will provide realistic basis and theoretical basis for further study the secondary metabolites of endophytic fungi associated with Ferula, and the relationship between endophytic fungi and their host plants.
Biodiversity
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Ecology
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Endophytes
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isolation & purification
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Ferula
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microbiology
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Fungi
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classification
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isolation & purification
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metabolism
4.Analysis of Chemical Components and Antifungal Activity of Extraction from Conidia of Trichoderma viride LTR-2
Kai CHEN ; He-Tong YANG ; Ji-Shun LI ; Jin-Dong HU ; Guang-Zhi ZHANG ;
Microbiology 1992;0(03):-
To study the chemical components and the antifungal activity of extraction from conidia of Trichoderma viride LTR-2.The extraction were obtained by distilling with Methylene dichloride from conidia of Trichoderma viride LTR-2 cultured on wheat bran solid matrix.Antifungal activity were determined by mycelium growth method.The chemical components of the extraction were analysed by GC-MS,the relative components in the extraction were determined by area normalization.The extraction not only have broad-spectrum control,showed antibiosis against eleven different plant fungal pathogens in PDA dish,such as Rhizoctonia solani,Alternaria brassica,Verticillium dahliae,Macrophoma kawatsukai,Fusarium moniliforme,Botrytis cinerea,Rhizoctonia cerealis,Fusarium oxysporum f.sp.vasinfectum,Bipolaris sorokinana,Fusarium graminearum,Alternaria.mali,but also have high inhibitory effect,and had 89.3% suppressive rate to Rhizoctonia cerealis.About sixty components were separated and identified by GC-MS,majority components were Hydrocarbon,the number of the Hydrocarbon were fourty-three kinds.Ergosterol was the major chemical components of the extract,and has 41.90% content.Other components comprised:Ketone,Organic acid,Alcohol,Ene,et al.Conclusion:The extraction from conidia of Trichoderma viride LTR-2 have antifungal activity.The extration comprised 2H-Pyran-2-one,5,6-dihydro-6-pentyl,it has 2.35% content.reference others literature,2H-Pyran-2-one,5,6-dihydro-6-pentyl may be the suppressive component of the extration.
6.Silencing of COX-2 in nasopharyngeal carcinoma cells with a shRNAmir lentivirus vector.
Gang LI ; Xiang-Ping LI ; Li JIANG ; Juan LU ; Xiong LIU ; Shun-Jin CHEN
Journal of Southern Medical University 2009;29(6):1111-1114
OBJECTIVETo construct a miR-155-based lentivirus vector to induce cyclooxygenase-2 gene silencing in nasopharyngeal carcinoma (NPC) cells by expressing anti-COX-2 shRNAmir.
METHODSmiR-155-based anti-COX-2 shRNAmir template was synthesized and inserted into pLVTHM plasmid. The recombinant pLVTHM/shRNAmir was transfected into 293FT cells for packaging the lentivirus vector. After infection with the lentivirus vector, the GFP-positive cells were screened by flow cytometry, and COX-2 mRNA level was detected by RT-PCR.
RESULTSRestriction digestion and DNA sequencing confirmed successful construction of the anti-COX-2 vector pLVTHM/shRNAmir. A subline of C666-1 cells was established after infection with the lentivirus vector, and the COX-2 expression in the cells was stably silenced.
CONCLUSIONThe shRNAmir lentivirus vector constructed may serve as an effective COX-2 inhibitor, which may facilitate future studies of gene therapy of NPC.
Cell Line, Tumor ; Cyclooxygenase 2 ; genetics ; Genetic Vectors ; genetics ; Humans ; Lentivirus ; genetics ; metabolism ; MicroRNAs ; genetics ; Nasopharyngeal Neoplasms ; genetics ; metabolism ; pathology ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection
7.ITF-2357 on inhibition myeloid leukemic cell lines cells proliferation in vitro and its mechanism.
Wen-juan YU ; Lei WANG ; Liang-shun YOU ; Chen MEI ; Qiu-ling MA ; Jie JIN
Chinese Journal of Hematology 2012;33(5):366-370
OBJECTIVETo explore the effect of ITF2357, a novel histone deacetylase (HDAC) inhibitor, on the growth, differentiation and apoptosis of acute myeloid leukemic (AML) cells and its mechanism.
METHODSAML cell lines kasumi-1 cells as a model for AML1-ETO positive, and THP1 cells for AML1-ETO negative, the leukemic cells proliferation was analyzed by MTT assay, expression of myeloid-specific differentiation antigen and cell cycle by flow cytometry, cell apoptosis by annexin V staining and flow cytometry. AML1-ETO, acetyl-histone, and caspase protein was analyzed by Western blot.
RESULTS0.5 µmol/L ITF2357 treatment significantly inhibited kasumi-1 cells proliferation, with the 48 h half inhibitory concentration (IC(50)) of 0.1 µmol/L. The initial inhibitory concentration of THP1 cell line was 5 µmol/L. ITF 2357 induced apoptosis of kasumi-1 cells in a time- and dose-dependent manner. A dose-dependent increase in early apoptosis occurred at 24 hours treatment and in late apoptosis at 48 hours treatment by ITF2357. Early apoptosis cells increased from (1.44 ± 1.52)% to (24.51 ± 5.79)%. Late apoptosis cells increased from (2.37 ± 2.8)% to (63.66 ± 1.56)%. ITF2357 induced AML1-ETO degradation by caspase-dependent pathway. 0.25 µmol/L ITF2357 induced a time- and dose-dependent increase in expression of myeloid cell surface protein CD13 and CD15. 5 µmol/L ITF2357 blocked the cells at G(0)/G(1) phase, G(0)/G(1) cells increased from (39.69 ± 6.56)% to (79.2 ± 6.51)% and s-phase cells declined from (60.12 ± 3.29)% to (18.97 ± 6.62)%. Kasumi-1 cells incubated with 0.5 µmol/L of ITF2357, AML1-ETO protein began to decrease at 24 hours and could hardly be detected at 96 hours. ITF2357 induced AML1/ETO degradation through a caspase-dependent mechanism. At the same time, acetylated H3 and H4 increased.
CONCLUSIONLow-dose HDAC inhibitor ITF2357 can effectively inhibit the AML cells proliferation, especially for AML1-ETO positive AML cells. It inhibits Kasumi-1 cells proliferation degradation of AML1-ETO protein expression, blocks the cells at G(0)/G(1) phase, and induces apoptosis and differentiation of the cells.
Acetylation ; Apoptosis ; drug effects ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Histone Deacetylase Inhibitors ; pharmacology ; Histones ; metabolism ; Humans ; Hydroxamic Acids ; pharmacology ; Oncogene Proteins, Fusion ; metabolism
8.Clinic application of skin flap based lateral supramalleolar branches of the peroneal artery.
Shun-bing WANG ; Fa-lin WU ; Sheng-hua NI ; Jian-hua HUANG ; Jian-feng CHEN ; Ding-jin SHAN
China Journal of Orthopaedics and Traumatology 2008;21(7):521-522
Adolescent
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Adult
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Aged
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Ankle Injuries
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surgery
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Female
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Foot Injuries
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surgery
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Humans
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Male
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Middle Aged
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Soft Tissue Injuries
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surgery
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Surgical Flaps
9.Deoxyribonuclease I gene expression in systemic lupus erythematosus patients.
Xue-bing FENG ; Nan SHEN ; Shun-le CHEN ; Jin HUA ; Li SUN ; Jie QIAN
Chinese Journal of Medical Genetics 2003;20(6):477-481
OBJECTIVETo observe whether deoxyribonuclease I (DNASE1) gene expression and its DNASE1 mRNA expression was detected by real-time polymerase chain reaction and its alternatively spliced transcripts were performed by capillary electrophoresis. An analysis was also made to disclose whether specific single nucleotide polymorphisms (SNPs) haplotype had effects onDNASE1 gene expression and its alternatively spliced transcripts.
RESULTSDNASE1 gene expression was higher in SLE patients than in normal controls (P<0.001), and in patients it was found to be of no relationship with SLE disease activity index score. However, it was increased in female patients. Capillary electrophoresis revealed that the pattern of alternatively spliced transcripts in patients was not the same as that in normal controls. Moreover, it seemed that different SNPs haplotype combination might show different transcript pattern in SLE patients.
CONCLUSIONIn SLE patients, DNASE1 gene expression is abnormal and there are alternatively spliced transcripts different from those in normal controls. DNASE1 gene is a critical factor in the pathogenesis of SLE.
Adolescent ; Adult ; Alternative Splicing ; Deoxyribonuclease I ; genetics ; Female ; Gene Expression ; Humans ; Lupus Erythematosus, Systemic ; genetics ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide
10.A case of Richter syndrome transformed from chronic lymphocytic leukemia with karyotype aberration of trisomy 12.
Zhen-Shu XU ; Jin-Yan ZHANG ; Rong ZHAN ; Zhi-Hong ZHENG ; Shun-Quan WU ; Zhi-Zhe CHEN
Journal of Experimental Hematology 2012;20(2):287-290
This study was aimed to investigate the relationship between Richter's syndrome (RS) transformation and clinical characteristics as well as karyotype of patient with chronic lymphocytic leukemia (CLL). By the follow-up of a patient with CLL, the clinical characteristics, karyotype, treatment pattern and its effect, as well as disease progression were monitored regularly with serological test, flow cytometry and FISH technique. The results indicated that the patient typically presented with history of CLL at initial diagnosis, with expression of CD5(+), CD19(+) and CD23(+), Binet stage C, as well as karyotype aberration of trisomy 12, and poorly responded to 4 cycles of standard chemotherapy of FCR regimen. The disease progression was confirmed at 5 months with the symptoms of fever in the absence of infection, elevated lactate dehydrogenase level and rapidly enlarging lymphnodes which showed typically diffuse large B cell lymphoma by the biopsy. It is concluded that karyotype aberration of trisomy 12 is one of the risk factors for RS transformation, and treatment pattern of the patient with CLL may be associated with the transformation of RS.
Cell Transformation, Neoplastic
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genetics
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Chromosomes, Human, Pair 12
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Female
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Humans
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Karyotype
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Karyotyping
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Leukemia, Lymphocytic, Chronic, B-Cell
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genetics
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Middle Aged
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Trisomy