Lgr5 is one of the G protein-coupled receptors families, the newest research displays that it has the potency as the special marker of stem cell, it also has a great relationship with tumor formation and development. Lgr5 may be a marker of tumor stem cells, and can be the new treatment target.

Objective To investigate the application of “top-down” method in assessment of measurement uncertainty of bio-chemical detection indicators .Methods The assessment data of internal quality control and external quality and “top-down”method were used to assess the precision and accuracy of 26 biochemical indicators ,and the two variables above were combined to calculate the measurement uncertainty .Results Uncertainty of 18 biochemical indicators was compatible with the target uncertainty ,ac-counting for approximately 69 .2% of all assessment projects .Five indicators ,such as TBIL ,albumin ,CK ,calcium and magnesium , was not compatible with the target uncertainty ,accounting for approximately 19 .2% of all assessment projects .Conclusion The“top-down” method is effective and feasible for assessment of measurement uncertainty of biochemical detection indicators .

Objective To investigate the distribution and resistance of clinical isolates obtained from the second affiliated hospi-tal of Chongqing medical university in 2012 .Methods The bacteria strains isolated from clinics were collected .Identification and drug susceptibility test were performed by automatic analysis system and API manual identification system .The date was analyzed according with software WHONET 5 .6 .Results A total of 3 454 bacterial isolates were obtained ,which included 36% gram-posi-tive strains ,64% gram-negative strains and 1% Anaerobic bacteria .The detection rates of methicillin-resistant S .aures was 33% , the detection rate of vancomycin -resistant enterococci was 1 .6% .In enterobacteriaceae ,ESBLs producing strains accounted for 60 .6% and 35 .8% in E .coil and K .pneumonia respectively .The drug resistance of A .baumannii and P .aeruginosa was increased . Conclusion Drug resistance of bacterial isolated from our hospital is universal .Drug monitoring data is important for clinical treat-ment .

Objective To investigate the inhibitory effects and mechanisms of a nature product derivate oridonin on in?vasion of human lung cancer. Methods Human lung cancer A549 and PC-9 cell lines were treated with oridonin. MTS as?say was used to determine cell proliferation. Transwell assay was used to determine the cell invasion, and adhesion assay to determine the cell adhesion. Flow cytometry was used to determine cell cycle. Western blotting and realtime-PCR were used to detect expression levels of CDK1, mTOR, p53, p21, E-cadherin, CD44,β-catenin, uPA, MMP-2/9, p-AKT and p-Src. The luciferase reporter assay was used to detect the NF-κB promoter activity. Results In vitro proliferation, invasion and adhesion of A549 and PC-9 cells were significantly inhibited by oridonin. The cell cycle was halted by G2/M phase, and ex?pressions of E-cadherin, p53 and p21 were promoted, while expressions of CDK1, mTOR, CD44,β-catenin, uPA, MMP-2/9, p-AKT and p-Src and promoter activity of NF-κB were down-regulated. Conclusion Oridonin is able to inhibit the in vitro invasion of human lung cancer A549 and PC-9 cell lines, which might be correlated with its abilities to regulate the ty?rosine kinase activity.

@@

OBJECTIVE:To establish a simple and rapid preparation technique of Rifampicin eye drops.METHODS:The calculating quantity of hydrochloric acid was used to dissolve Rifampicin,then the equimol quantity of potassium hydroxide was added to neutralize the acid and yield potassium chloride,whose quantity was designed according to the prescrip?tion.RESULTS:Neutralization reaction not only overcame the difficulty of Rifampicin's dissolution in water,but also avoided the irritation to eyes caused by using ethanol as solvent.CONCLUSION:The method is well-versed in design,simple in preparation and controllable in quality.

Objective: To establish a quality control standard for YurongXiaocuo granule.Methods: TLC was used to identify Radix Puerariae,Fructus Forsythiae,Bulbus Fritillariae Thunbergii,while the content of tanshinoneⅡA was determined by HPLC.Result: The methods of qualitation and quantitate was simple,accurate and with strong specifi city,the good linear rang of tanshinoneⅡA was 3.2-32?g and the average recovery was 99.51%(RSD=2.74%).Conclusion: This method can control effectually the quality of YurongXiaocuo granule.

Objective To construct a stable infectious clone of S191 virus strain used for the pro-duction of live attenuated measles vaccine. Methods Full length cDNA of S191 strain and gene fragments encoding nucleocapsid(N),phosphoprotein(P)and RNA polymerase(L)were synthesis and respectively cloned into the vector pVAX1. The 293T cells were respectively transfected with the recombinant expression plasmids and co-cultured with Vero cells. The supernatants of cell culture were collected for identifying res-cued viruses. The indirect immunofluorescence assay was performed for virus identification. The rescued viruses at different passages in Vero cells and the sequences of the rescued viruses were analyzed. Results Restriction enzyme digestion and sequence analysis showed that the recombinant expression plasmids contai-ning the full length cDNA with an artificially engineered mutation at nucleotide 2101(C-A)and gene frag-ments encoding N,P and L proteins of S191 strain were constructed successfully. The N and P proteins were detected in Vero cells with immunofluorescence assay. A cytopathogenic effect on Vero cells was induced by rescued viruses. Conclusion The stable infectious clones of S191 virus used for the production of live at-tenuated measles vaccine were rescued successfully. An approach by using reverse genetics technique for S191 strain study was established which could be used for the development of new chimeric vaccines against measles virus.

Objective To study a rehabilitation nursing menus for neurogenic large intestine dysfunction.Methods Various nursing approaches were used for defecation dysfunction.Results and Conclusion 94.60% patients improved in the second week,which including gained more awareness,control,and spend less time of defecation.

Objective: To evaluate the effectiveness of droplet digital PCR (ddPCR) assay for detection of neuraminidase (NA) H275Y mutations in influenza A H1N1 pdm09 virus. Methods: The primers and dual probes were designed based on the sequence of the H1N1 pdm09 NA gene fragment which contained 275 amino acid sites, and the annealing temperature of ddPCR assay was optimized to establish a method for detection of H275 drug-sensitive genes and H275Y drug-resistant genes in H1N1 pdm09 virus. The sensitivity of ddPCR assay and fluorescent quantitative PCR (qPCR) assay was compared using the detection limit, and the specificity of ddPCR and qPCR assays was compared for detection of 14 respiratory virus samples. In addition, 64 clinical samples and 5 influenza isolates were tested to calculate the abundance of H275Y mutations, and the mutation abundance of 5 influenza isolates was compared with next-generation sequencing results. Results: The optimal annealing temperature was 62.2 ℃. The detection limits of ddPCR assay were 5.28 (95%CI: 4.28-7.45) copies/reaction for H1N1 pdm09 H275 drug-sensitive plasmids and 6.51 (95%CI: 5.25-9.37) copies/reaction for H1N1 pdm09 H275Y drug-resistant plasmids, and the detection limits of qPCR assay were 5.70 (95%CI: 4.83-7.45) copies/reaction for H1N1 pdm09 H275Y drug-sensitive plasmids and 7.06 (95%CI: 5.92-9.40) copies/reaction for H1N1 pdm09 H275Y drug-resistant plasmids. Both ddPCR and qPCR assays detected H275 and H275Y drug-resistant plasmids in H1N1 pdm09 viral samples but did not detect H275 and H275Y drug-resistant plasmids in other 11 respiratory virus samples, and these two assays showed consistent results. Of the 64 clinical samples, ddPCR assay detected H275Y mutation in three pharyngeal swab specimens from a severe pneumonia patients infected with H1N1 pdm09 virus, and the greatest mutation abundance was detected in samples collected on day 4 post-treatment with oseltamivir phosphate (53.37%). ddPCR assay detected 0.63, 88.93%% and 1.27% H275Y mutation abundance in samples collected on days 2, 4 and 5 post-treatment with oseltamivir phosphate, and next-generation sequencing detected 89.46% H275Y mutation abundance in samples collected on day 4 post-treatment with oseltamivir phosphate; however, no H275Y mutation was detected in samples collected on days 2 or 5 post-treatment with oseltamivir phosphate. Conclusions ddPCR presents a higher sensitivity and specificity than qPCR assay for detection of H275Y mutations in H1N1 pdm09 virus, and presents a higher sensitivity than next-generation sequencing for detection of low-frequency mutations, which is effective for quantitative detection of H275Y mutations in the NA fragment of the H1N1 pdm09 virus.