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.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Injections, Intravenous ; Male ; Middle Aged ; Motor Neuron Disease ; drug therapy ; Phytotherapy ; Treatment Outcome

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.

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 ; Ecology ; Endophytes ; isolation & purification ; Ferula ; microbiology ; Fungi ; classification ; isolation & purification ; metabolism

Animals ; Cell Culture Techniques ; Cells, Cultured ; Male ; Neural Stem Cells ; cytology ; Neurons ; cytology ; Olfactory Mucosa ; cytology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo separate the cell subpopulation with high tumorigenic ability and study the biological characteristics of this subpopulation in laryngeal carcinoma cells.

METHODSHuman laryngeal carcinoma cells were obtained by primary tissue culture technique. CD44 and CD133 molecules were used as markers to isolate the CD44(+), CD133(+), CD44(+)CD133(+) and CD44(+)CD133(-) cell subpopulations from the laryngeal carcinoma cells by flow cytometry. A nude mouse tumor xenograft model was developed for the study of the tumorigenic effects of the different cell populations. 1 x 10(6), 1 x 10(5), 1 x 10(4) and 1 x 10(3) cells were injected into the left flank of the mice, respectively. The mice were observed for palpable tumor formation and were sacrificed at 4 weeks later to assess the tumor formation rate, tumor volume and tumor weight. Boyden chamber migration assay was used to determine the migration ability and immunochemistry was used to detect the expression of stem cell antigen SCA-1 and beta1-integrin. Semi-quantities RT-PCR and Western blot analysis were performed to detect the expression level of Bmi-1 in the different cell subpopulations.

RESULTSThe growth of subcutaneous tumors in nude mice showed that a tumor can be generated with 1 x 10(3) CD44(+)CD133(+) cells. When the same dose of 1 x 10(6) CD44(+)CD133(+) cells was injected into the mice, both the average weight and volume of the tumors were significantly higher than those generated from other cell subpopulations. Boyden chamber migration assay showed that the invasion ability of CD44(+)CD133(+) cells was significantly higher than that of other cell subsets. The results of immunochemical analysis showed an abundant expression of stem cell antigen SCA-1 and beta1-integrin in the CD44(+)CD133(+) cells. Semi-quantitative RT-PCR and Western blot analysis provided strong evidence that the Bmi-1 expression in CD44(+)CD133(+) and CD133(+) cells was very significantly higher than that in CD44(+), CD44(+)CD133(-) and control cells (P < 0.01).

CONCLUSIONOur findings demonstrate that CD44(+)CD133(+) subset cells in laryngeal carcinoma posses some biological characteristics of tumor stem cells, which may be the original cells of laryngeal carcinoma and may become a new target of tumor therapy.

AC133 Antigen ; Animals ; Antigens, CD ; analysis ; Antigens, Ly ; metabolism ; Cell Adhesion ; Gene Expression Regulation, Neoplastic ; Glycoproteins ; analysis ; Humans ; Hyaluronan Receptors ; analysis ; Integrin beta1 ; metabolism ; Laryngeal Neoplasms ; immunology ; metabolism ; pathology ; Male ; Membrane Proteins ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Nuclear Proteins ; genetics ; metabolism ; Peptides ; analysis ; Polycomb Repressive Complex 1 ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA ; metabolism ; Repressor Proteins ; genetics ; metabolism ; Tumor Burden ; Tumor Cells, Cultured

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 ; genetics ; Chromosomes, Human, Pair 12 ; Female ; Humans ; Karyotype ; Karyotyping ; Leukemia, Lymphocytic, Chronic, B-Cell ; genetics ; Middle Aged ; Trisomy

OBJECTIVETo discuss the individual operation standard and safety on the deep puncturing at Tianshu (ST 25).

METHODSThe deep insertion through the peritoneum about 1-2 millimeter was taken as individual standard depth at Tianshu (ST 25) for 39 patients. Using CT cross-section scanning and image surveying, the regression analysis was conducted on the impacts of abdominal circumference (AC,cm) to the peritoneum depth.

RESULTSIt was concluded the formula for calculating the satisfactory depth of individual insertion at Tianshu (ST 25), depth = -0.562 + 0.045 x AC.

CONCLUSIONThe depth of insertion, 1-2 millimeter through peritoneum at Tianshu (ST 25) is safe and feasible in acupuncture treatment, which can be used significantly as the reference and guidance for the assessment of clinical operation and safety on the deep puncturing of the other acupoints on the abdominal region.

Acupuncture Points ; Acupuncture Therapy ; adverse effects ; methods ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Regression Analysis ; Tomography, X-Ray Computed

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

BACKGROUNDImmunological sensitization remains a major problem following renal transplantation. There is no consensus for the management of sensitized renal allograft recipients. The patients become tethered to dialysis while waiting for compatible donors. This study was designed to evaluate the efficacy and safety of preoperative single-bolus high-dose antithymocyte globulin (ATG) as induction therapy in sensitized renal transplant recipients.

METHODSA total of 56 patients were divided into two groups according to the level of panel reactive antibody (PRA): non-sensitized group (PRA < 10%, n = 30) and sensitized group (PRA > or = 10%, n = 26). The characteristics of the recipients and donors were comparable between the two groups. Mycophenolate mofetil (MMF, 1 g) or ATG (iv. 9 mg/kg) were given preoperatively in the two groups as induction therapy. After the transplantation, the patients were treated with standard triple therapy regimen consisting of tacrolimus (FK-506) or cyclosporine A, MMF, and prednisolone. Acute rejection (AR) and infection episodes were recorded and renal function was monitored during a 12-month follow-up. Chi(2) test and t test were used to analyze the data.

RESULTSDuring the follow-up, 6 patients (20.0%) suffered AR episodes in the non-sensitized group and 4 (15.4%) in the sensitized group (P = 0.737); 8 patients (26.7%) experienced 11 infection episodes (average, 1.4 episodes per infected patient) in the non-sensitized group, and 6 (23.1%) experienced 10 infection episodes (average, 1.7 episodes per infected patient) in the sensitized group (P = 0.757, 0.890). The safety of the drugs, which was assessed by the occurrence of side effects, was comparable between the two groups. The hospital stay was 13 - 25 days (mean, 16.7 +/- 3.3) in the non-sensitized group and 14 - 29 days (mean, 16.2 +/- 3.1) in the sensitized group, respectively (P = 0.563). No delayed graft function (DGF) was observed in all the patients. Both the 12-month actuarial patient and graft survival rates were 100% in the two groups.

CONCLUSIONPreoperative single-bolus high-dose ATG is an effective and safe induction therapy yielding acceptable acute rejection rate in sensitized renal transplant recipients.

Adult ; Antilymphocyte Serum ; adverse effects ; therapeutic use ; Female ; Graft Rejection ; prevention & control ; Graft Survival ; Humans ; Immunosuppressive Agents ; Kidney Transplantation ; Male ; Middle Aged