Objective To summarize the experiences of treatment of calculus obstructive pyonephrosis with transureteroscopic holium laser lithotripsy. Methods There were 13 patients with flank pain and fever, all patients received antibacterial therapy and nutritional support before operation ,the patients of calculus obstructive pyonephrosis were treated using transureteroscopic holium laser lithotripsy. Results The ureteral calculus were cleared off in 10 cases, and in three cases, the stone moved to pelvis during operation that they needed additional procedure of ESWL, no severe complications were observed. Conclusion Transureteroscopic holium laser lithotripsy is safe,minimal in-vasive and effective method for treatment of calculus obstructive pyonephrosis and the therapeutic scheme can be the first choice.

Objective To observe the expression of aggrecanase 2 and a tissue inhibitor of metalloproteinase 3 (TIMP-3) in degenerate human lumbar intervertebral discs and their role in degeneration of the nucleus pulposus.Methods Pfirrmann classification was used to class degenerate intervertebral discs observed through MRI.They were divided into three groups:a control group (Pfirrmann grade Ⅰ-Ⅱ),a degeneration group (Pfirrmann grade Ⅲ-Ⅳ),and a severe degeneration group (Pfirrmann grade Ⅴ).A total of 45 cases accepted lumbar spine surgery for removing nucleus pulposus specimens.Each group contained 15 cases.After formalin-fixation and paraffin embedding,immunohistochemistry was used to detect aggrecanase 2 and TIMP-3 expression in the nucleus pulposus cells.Results The percentages of cells positive for aggrecanase 2 were (13.58 ± 7.76) ％,(33.48 ± 13.95) ％ and (56.00 ± 18.39) ％ in the control,degeneration and severe degeneration groups respectively.These differences had statistical significance.The percentages of cells positive for TIMP-3 were (34.78 ± 13.80) ％,(46.77 ± 10.98) ％ and (50.65 ± 16.45) ％,and these differences were again statistically significant.The aggrecanase 2/TIMP-3 ratios were also significantly different.Conclusion As the degree of degeneration of the nucleus pulposus increased,the expression of aggrecanase 2 and TIMP-3 rose,which indicates that both changes were closely connected with the degeneration.Their ratio was correlated with the degree of degeneration of the nucleus pulposus.

Objective To isolate and purify human coagulation factor Ⅶ from Cohn fraction Ⅲ precipitate.Methods The purification procedure of human factor Ⅶ from Cohn fraction Ⅲ precipitate involves dissolving fraction Ⅲ,absorbing factor Ⅶ onto barium citrate and eluting,ammonium sulfate fractionation and DEAE-Sepharose Fast Flow ion exchange chromatography,and Sephadex G-100 gel filtration chromatography.Results 10.1mg purified FⅦ was obtained from 400g Cohn fraction Ⅲ precipitate.The purified FⅦ has a specific clotting activity of 1775.8U/mg and the overall yield of FⅦ specific clotting activity is 17.6% of the starting material.The purity of FⅦ was judged by SDS-PAGE and there was only one protein band on the gel.Conclusion The procedure of purifying Ⅶ from Cohn fraction Ⅲprecipitate is established with satisfactory purity.

Objective To analyze the genotype and plasmid transfer of Enterobacteriaceae carring blaNDM‐1 with blaIMP‐4 or blaKPC‐2 .Methods From April 2012 to October 2014 ,a total of 33 non‐repeatitive carbapenem‐resistant Enterobacteriaceae ( including Imipenem‐resistant , meropenem‐resistant or Ertapenem‐resistant) were isolated from 5 hospitals in Wenzhou and Hangzhou . Identification and antimicrobial susceptibility test were performed using Vitek 2 Compact automatic microbiology analyzer . Phenotypes of carbapenemase were screened using modified Hodge test and ethylenediamine tetraacetic acid‐disk synergy test .Extended spectrum βlactamase test was determined by the double disk combination test which was recommended by Clinical and Laboratory Standards Institute .AmpC activity was tested by a three‐dimensional Cefoxitin method .Drug resistant genes including blaNDM‐1 and linkage of ISAba125‐NDM were detected by polymerase chain reaction (PCR) .The purified PCR products were cloned and sequenced .Plasmid conjugation experiment and elimination method were carried out to test partial bacterial strain and K . pneumoniae carrying blaNDM‐1 with blaIMP‐4 or blaKPC‐2 .Results Of the 33 non‐repeatitive carbapenem‐resistant Enterobacteriaceae ,28 were strains of K .pneumoniae ,1 strain of K . oxytoca,2strainsof Escherichiacoli,1strainof K.planticolaand1strainof E.cloacae.Thirteenstrains were isolated from Hospital of Sir Run Run Shaw of Zhejiang University ,thirteen from Wenzhou Hospital of Traditional Chinese Medicine ,one from Wenzhou People′s Hospital ,three from the First Affiliated Hospital of Wenzhou Medical University and three from Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine .Thirty‐one strains were confirmed as carbapenemase‐producing with 24 of blaKPC‐2 ,3 of blaNDM‐1 ,1 of blaNDM‐5 and 3 of blaIMP‐4 .Among them ,one strain carried blaNDM‐1 with blaIMP‐4 and one strain carried blaNDM‐1with blaKPC‐2 ,respectively .The plasmid transfer and conjugation experiment was performed between strains carrying blaNDM‐1 and Escherichia coli EC600 or K . pneumoniae ATCC13833 and genes of blaNDM‐1 and ISAba125‐NDM were obtained .Conclusions blaKPC‐2 gene is the popular carbapenemase genotype .blaNDM‐1 or blaNDM‐5 may be correlated with linkage gene of ISAba125‐N DM .Coexistence of blaNDM‐1 carrying blaIMP‐4 or blaKPC‐2 is detected in the same strain , respectively . Enough importance should be attached to the strains ,because most of them are multiple drug resistance with related genes located in the plasmid which is easily spread between strains .

TCP family as plant specific transcription factor, plays an important role in different aspects of plant development. In order to screen TCP family members in tobacco, the homologous sequences of tobacco and Arabidopsis TCP family were identified by genome-wide homologous alignment. The physicochemical properties, phylogenetic relationships and cis-acting elements were analyzed by bioinformatics. The homologous genes of AtTCP3/AtTCP4 were screened, and RT-qPCR was used to detect the changes of gene expression upon 20% PEG6000 treatment. The results show that tobacco contains 63 TCP family members. Their amino acid sequence length ranged from 89 aa to 596 aa, and their protein hydropathicity grand average of hydropathicity (GRAVY) ranged from -1.147 to 0.125. The isoelectric point (pI) ranges from 4.42 to 9.94, the number of introns is 0 to 3, and the subcellular location is all located in the nucleus. The results of conserved domain and phylogenetic relationship analysis showed that the tobacco TCP family can be divided into PCF, CIN and CYC/TB1 subfamilies, and each subfamily has a stable sequence. The results of cis-acting elements in gene promoter region showed that TCP family genes contain low docile acting elements (LTR) and a variety of stress and metabolic regulation related elements (MYB, MYC). Analysis of gene expression patterns showed that AtTCP3/AtTCP4 homologous genes (NtTCP6, NtTCP28, NtTCP30, NtTCP33, NtTCP42, NtTCP57, NtTCP63) accounted for 20% PEG6000 treatment significantly up-regulated/down-regulated expression, and NtTCP30 and NtTCP57 genes were selected as candidate genes in response to drought. The results of this study analyzed the TCP family in the tobacco genome and provided candidate genes for the study of drought-resistance gene function and variety breeding in tobacco.
Nicotiana/genetics* ; Phylogeny ; Plant Breeding ; Amino Acid Sequence ; Arabidopsis ; Polyethylene Glycols