Objective To analyze the infection sites of pseudomonas aeruginosa (PAE) isolated from patients in hospital ,and in‐vestigate their drug resistance situation ,in order to provide reference information for clinical use of antibiotics rationally .Methods The sample distribution of PAE between January 2012 and December 2013 were retrospectively analyzed .And the resistance rates of PAE to antibacterial drugs from different sites of patients were statistically compared .Results The isolation rate of PAE in re‐spiratory tract was the highest ,accounting for 74 .1% ,closely followed by isolation rate in urine and wound secretion .The resist‐ance rate of PAE to antibacterial drugs in these three kinds of specimen is statistically different (P<0 .05) .The resistance rate of PAE is high in respiratory tract ,and low in wound secretion .Conclusion The pseudomonas aeruginosa infection is mostly common‐ly found in respiratory tract ,and has the highest drug resistance rate .The choice of antibacterial drug should be made according to the infection sites of patients ,because the resistance rate of PAE in different sites of patients is significantly different .

OBJECTIVETo establish a human embryonic stem cell line with stably β-catenin gene silencing by lentivirus-mediated shRNA interference.

METHODSPLKO.1-puro-β-catenin vector, a lentivirus plasmid expressing β-catenin shRNA, was packaged into 293FT cells. Human embryonic stem cells were infected with the lentivirus and the cell clones stably expressing β-catenin shRNA were selected by puromycin, with the uninfected cells and cells infected with the empty vector as the control. Real-time RT-PCR was used to evaluate the efficiency of β-catenin knocked down; β-catenin and OCT4 protein expression in the infected cells was examined using immunofluorescence assay.

RESULTSInfection with β-catenin-specific shRNA lentivirus resulted in stable interference of β-catenin expression in human embryonic stem cells, which showed a β-catenin mRNA expression of only 1% of that in the uninfected cells. Infection with the empty vector produced no obvious effect on β-catenin expression compared to the uninfected cells. In the cells infected with β-catenin shRNA lentivirus, β-catenin protein expression was almost undetectable in immunofluorescence assay, while OCT4 was still expressed after the interference.

CONCLUSIONLentiviral vector-delivered shRNA results in effective and stable β-catenin gene silencing in human embryonic stem cells.

Cell Line ; Embryonic Stem Cells ; cytology ; metabolism ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; Plasmids ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; Wnt Signaling Pathway ; beta Catenin ; genetics ; metabolism

Objective Screening the immune polypeptide sequence of toxoplasma (Tg) CDPK5 gene ,which were synthesized and then immunized the New Zealand white rabbit to prepare antiserum ,and identification its function .Methods Bioinformatics a‐nalysis was used to determine the immune peptide of Tg CDPK5 sequence ,which were artificially synthesized to immune white rab‐bit to prepare antiserum .The titers of antibodies were determined by ELISA and the polyclonal antibodies were verified with CD‐KP5 antigen by Western blot .The sub‐cellular localization of Tg CDPK 5 were obtained by immunofluorescence assay .Results 17 bp peptide sequence from the Tg CDPK5 N‐terminal were chosen as immune polypeptide by bioinformatics analysis .Synthetic pep‐tide were used to immune rabbit to obtain polyclonal antiserum .The result showed that the titer of the obtained ployantibody were 1∶640 000 ;Western blot demonstrated that the antiserum could specifically recognize Tg CDPK 5(75 .4 × 103 );Immunofluores‐cence assay revealed this antibody could specifically recognize the endogenous Tg CDPK 5 of Toxoplasma gondii .Conclusion Ac‐cording to the analysis of Tg CDPK5 sequence information ,this study successful obtained Tg CDPK5 polyclonal antibody .

The widespread application of next generation sequencing (NGS) in clinical settings has enabled testing, diagnosis, treatment and prevention of genetic diseases. However, many issues have arisen in the meanwhile. One of the most pressing issues is the lack of standards for reporting genetic test results across different service providers. The First Forum on Standards and Specifications for Clinical Genetic Testing was held to address the issue in Shenzhen, China, on October 28, 2017. Participants, including geneticists, clinicians, and representatives of genetic testing service providers, discussed problems of clinical genetic testing services across in China and shared opinions on principles, challenges, and standards for reporting clinical genetic test results. Here we summarize expert opinions presented at the seminar and report the consensus, which will serve as a basis for the development of standards and guidelines for reporting of clinical genetic testing results, in order to promote the standardization and regulation of genetic testing services in China.