Objective To discuss the value in quality improvement and continuous improvement through the way for determina-tion of target uncertainty in chemical quantitative detection project and regular evaluation of measurement uncertainty in different phase.Methods Based on the biological variability of quality specification and CNAS-TRL-001:CNAS technical report,to compare the five kinds of determination of target uncertainty.Method 1:the fundamental way(calculate the different levels of target impre-cision and bias);Method 2:biological variation of different grades of total allowable error;Method 3:the relative expanded uncer-tainty evaluation value based on target imprecision and bias;Method 4:the target relative expanded uncertainty based on biological variation of different grades;Method 5:the target relative expanded uncertainty based on the quality index of analysis.We used un-certainty evaluate index(UEI)to review the changes of uncertainty in different phase.Results The 14 conventional chemistry tests in 2013 with metrological traceability and participate in the Ministry of health EQA were as the target.There was no significant difference among the 2,3,4 method,the ratio of which reached the ideal value of uncertaninty target were not significantly different. In method 5,9 projects achieved the target of uncertainty requirements,accounted for 64.3%.TP,ALT,BUN,UA,CK,these5 pro-jects′UEI were less than 0,accounted for 35.7%;other 9 projects′UEI were more than 2.0%.Conclusion Method 5:the target relative expanded uncertainty based on the quality index of analysis which is based on WS/T403-2012 can give consideration to the quality standard of repeatability precision and bias in the laboratory at the same time,and is easy to be accepted for laboratory;method 4:the target relative expanded uncertainty based on biological variation of different grades is in the same way with the eval-uation of test results uncertainty,is better than method 2 and 3;method 1 is the fundamental way,can give the specific reasons when the test results cannot get the target uncertainty.Use UEI to assess the changes of uncertainty in different phase is more sensitive to changes of the test results′accuracy and its usefulness needs to be confirmed in practice.

Objective To explore the differences of using different analytic quality requirements in the comparable validation of blood cell analysis multi-system range test comparable schemes for establishing appropriate analysis quality standards for laboratory.Methods According to WS/T 407-2012 Guideline for Comparability Verification of Quantitative Results in Medical Institution,the range test comparable method was established.According to different sources of analytic quality requirements from the WS/T 406-2012 Analysis Quality Requirements of Clinical Hematological Detection Routine Items,the US Clinical Laboratory Improvement Amendment (88),GB/T 20470-2006 Requirements of External Quality Assessment for Clinical Laboratories and biological variations,corresponding analysis quality requirements standard was designed.Results With the standards designed by using WS/T 406-2012,CLIA′-88 and GB/T 20470-2006 as the analysis quality requirements,only the comparison results of low concentration levels in 3 items of HBC,PLT and HCT were not passed,while other results all were passed;all results passed the consistency verification by suitably revising the analytic quality requirements of low value concentrations.With the biological variations as the analysis quality requirement,the comparison results in WBC three concentration levels,and HBG high and low concentration levels were passed,but other results were not passed.Conclusion The biological variations analytical quality requirements are relative demanding.Using WS/T 406-2012 Analysis Quality Requirements of Clinical Hematological Detection Routine Items and GB/T 20470-2006 Requirements of External Quality Assessment for Clinical Laboratories,fully considering the suitability of low concentration quality requirements and formulating appropriate analysis quality standards of laboratory are the important contents of laboratory comparable validation scheme.

BACKGROUND:It is reported that tailless-like protein (TLX) plays critical roles in the regulation of early developmental processes in vertebrates, and it plays a key role in stem cells proliferation and differentiation into neurons. OBJECTIVE: To construct recombinant plasmid pEGFPN1-TLX and study the transfection into dermal multipotential stem cells. DESIGN, TIME AND SETTING: Cytogene experiment was performed at the Department of Pathogen Biology, School of Basic Medical Science, the Third Military Medical University of Chinese PLA from March to December 2007. MATERIALS: An adult SD was obtained from the Experimental Animal Center of the Third Military Medical University of Chinese PLA; dermal moltipotential stem cells (DMSCs) were cultured by the Institute of Combined Injury of the Third Military Medical University of Chinese PLA; pEGFPN1 and DH5α was gifted by professor Xu.METHODS: Total RNA was extracted from rat brain tissue to amplify TLX-coded cDNA sequence using RT-PCR. T/A was cloned on pMD18-T vector and determined using BamHI and Hindlll. The products were positive recombinant plasmid pMD18-T-TLX segments, which were sub-cloned in pEGFPN1 to construct recombinant plasmid pEGFPN1-TLX. Finally, pEGFPN1-TLX was transfected into DMSCs.MAIN OUTCOME MEASURES: The fluorescence protein expression was observed under fluorescence microscope at 24 hours after transfection; TLX mRNA expression was detected using RT-PCR; neuronal differentiation was observed using immunohistochemical staining.RESULTS: TLX full length cDNA was successfully cloned into pEGFPN1, and pEGFPN1-TLX was successfully constructed by means of sequence analysis and enzyme cutting identification. As compared with non-transfected DMSCs, pEGFPN1-TLX transfected DMSCs were observed after 10 days, formed resistant clones after 15 days, and shown a green fluorescent protein expression. However, non-transfected DMSCs died at day 10. RT-PCR indicated that pEGFPN1-TLX transfected DMSCs could express TLX mRNA. At day 3 after induction, NF200 positive cells were increased, but glial fibrillary acidic protein positive cells were decreased after induction of pEGFPN1-TLX transfected DMSCs.CONCLUSION: TLX was successfully constructed and transfected into DMSCs. After transfection, neuronal differentiation of DMSCs was enhanced, and the differentiation to gliocytes was inhibited.

In this study, apoptosis was induced by new type gosling viral enteritis virus (NGVEV) in experimentally infected goslings is reported in detail for the first time. After 3-day-old goslings were orally inoculated with a NGVEV-CN strain suspension, the time course of NGVEV effects on apoptotic morphological changes of the internal tissues was evaluated. These changes were observed by histological analysis with light microscopy and ultrastructural analysis with transmission electron microscopy. DNA fragmentation was assessed with a terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay and DNA ladder analysis. A series of characteristic apoptotic morphological changes including chromatin condensation and margination, cytoplasmic shrinkage, plasma membrane blebbing, and formation of apoptotic bodies were noted. Apoptosis was readily observed in the lymphoid and gastrointestinal organs, and sporadically occurred in other organs after 3 days post-infection (PI). The presence and quantity of TUNEL-positive cells increased with infection time until 9 days PI. DNA extracted from the NGVEV-infected gosling cells displayed characteristic 180~200 bp ladders. Apoptotic cells were ubiquitously distributed, especially among lymphocytes, macrophages, monocytes, and epithelial and intestinal cells. Necrosis was subsequently detected during the late NGVEV-infection phase, which was characterized by cell swelling, plasma membrane collapse, and rapidly lysis. Our results suggested that apoptosis may play an important role in the pathogenesis of NGVE disease.
*Adenoviridae/classification/pathogenicity ; Adenoviridae Infections/pathology/*veterinary/virology ; Animals ; *Anseriformes ; *Apoptosis ; Bird Diseases/*virology ; DNA Fragmentation ; Enteritis/*veterinary/virology ; Epithelial Cells/cytology/virology ; In Situ Nick-End Labeling ; Intestines/cytology/virology ; Leukocytes/cytology/virology ; Lymphoid Tissue/cytology/virology ; Macrophages ; Microscopy, Electron, Transmission