Objective To map the deletion breakpoint of 9p21 in breast cell line MCF-7 use the inverse PCR .Methods After di-gestion ,ligation and PCR reaction ,the breakpoint was confirmed by sequencing .Results The deletion breakpoint started at chr9 :21819532 and ended at chr9 :21989622 with a small insertion of ACTGG ,which was consistent with the result confirmed by the long range PCR .Conclusion The inverse PCR is one good method for deletion breakpoint mapping and suitable for large sample size detection .

ostic sensitivity and specificity of EUS are high in distinguishing benign and malignant character of upper digestive tract GIMTs. EUS plays an important role in guiding the clinical management of upper digestive tract GIMTs.

Caliciviridae Infections ; epidemiology ; China ; epidemiology ; Disease Outbreaks ; Female ; Humans ; Male ; Norovirus ; Schools

Child, Preschool ; China ; epidemiology ; Enterovirus A, Human ; genetics ; Female ; Hand, Foot and Mouth Disease ; epidemiology ; Humans ; Infant ; Male ; Molecular Epidemiology

In an effort to generate a desired expression construct for making heart-specific expression transgenic zebrafish, a Tol2 plasmid, which can drive EGFP reporter gene specifically expressed in the heart, was modified using subcloning technology. An IRES fragment bearing multiple cloning site (MCS) was amplified directly from pIRES2-EGFP plasmid and was inserted between the CMLC2 promoter and EGFP fragment of the pDestTol2CG vector. This recombinant expression plasmid pTol2-CMLC2-IRES-EGFP can drive any interested gene specifically expressed in the zebrafish heart along with EGFP reporter gene. To test the effectiveness of this new expression plasmid, we constructed pTol2-CMLC2-RED-IRES-EGFP plasmid by inserting another reporter gene DsRed-Monome into MCS downstream of the CMLC2 promoter and injected this transgenic recombinant plasmid into one-cell stage embryos of zebrafish. Under fluorescence microscope, both the red fluorescence and the green fluorescence produced by pTol2-CMLC2-RED-IRES-EGFP were detected specifically in the heart tissue in the same expression pattern. This novel expression construct pTol2-CMLC2-IRES-EGFP will become an important tool for our research on identifying heart development candidate genes' function using zebrafish as a model.
Animals ; Animals, Genetically Modified ; genetics ; growth & development ; DNA Transposable Elements ; genetics ; Genes, Reporter ; genetics ; Genetic Vectors ; biosynthesis ; genetics ; Green Fluorescent Proteins ; genetics ; Myocardium ; metabolism ; Plasmids ; genetics ; Transfection ; Transgenes ; Transposases ; genetics ; Zebrafish ; genetics ; Zebrafish Proteins ; genetics ; metabolism

Objective To explore the pulse oxygen saturation instrument in the application of vaginal delivery newborn, and analyze the pulse oxygen saturation in the clinical application value of vaginal delivery newborn. Methods From May 2012 to May 2013 in our hospital, 100 cases was born of the gestational age between 37 weeks to 42 weeks of vaginal delivery term newborns were randomly selected, and randomly divided into two groups in digital method: ob-servation group and control group, 50 cases in each group, after birth, the observation group with pulse oxygen satu-ration meter records the pulse of the fetus within 1 h each time values,and blood oxygen saturation, the control group was using Apgar score record the pulse of the fetus within 1 h each time values, and blood oxygen saturation, then put the two methods respectively recorded data entry into SPSS19.0 statistical software for analysis processing, analysis and comparison of pulse oxygen saturation and Apgar score in vaginal delivery was how to apply in the newborn and appli-cation value. Results Neonatal pulse oxygen saturation in observation group 1 min-monitoring value, 5 min-the moni-toring value of 10 min-monitoring value, 30 min-monitoring values were significantly higher than that of control group, significant difference was found in two groups, with statistical significance(P<0.05); Newborns in observation group 60 min-pulse oxygen saturation monitoring values similar to the control group, no significant differences between the two groups, no statistically significant (P>0.05);Neonatal pulse monitoring value in observation group, 1 min value, pulse pulse value when the 5 min,when 10 min pulse value, pulse value when 30 min, when 60 min pulse value were significantly higher than that of control group, significant difference, the two groups have statistical significance (P<0.05). Conclusion Pulse oxygen saturation meter compared with traditional Apgar score detection method, pulse oxy-gen saturation meter advantage in the application of vaginal delivery neonate, and reliability is extremely strong, ap-plication prospect, is worthy of popularization and application in clinical work.

Objective To obtain information on viral molecular structure and genome via full-length genomic analysis on the norovirus strain GZ20133135 isolated from Guangzhou.Methods Primers were designed according to the Sydney2012 full sequence.The full genome of the strain GZ20133135 was amplified by RT-PCR.The whole genome sequence was analyzed after cloned and sequenced.Results The genome of G Ⅱ-4 norovirus strain GZ20133135 consisted of 7566 bp,and it was revealed that there were three ORFs composites ORF1 (5100 bp),ORF2 (1623 bp),ORF3 (807 bp) respectively; ORF1 and ORF2 had 19 nt overlap.It was found by evolutionary comparative analysis that GZ20133135 genomic nucleotide sequences,compared with reference strains of G Ⅱ-4 Sydney2012 strains,the highest homology with a total length of homology was 99.07％.Phylogenetic analyses showed GZ20133135 belonged to G Ⅱ-4Sydney 2012 variant.Data of 541 amino acid analyses showed that Sydney 2012 variant strains of popular sites were aa294V or A or P→T,aa296S or T→S,aa297H or Q→R,aa298D or N or T→N,aa368T or N or S or A→E,aa372 N or S→D,aa393 N or D or S,aa394 T or G or S→T,aa395T or A→T,aa407 N or D→S,aa412T or D→N,aa413G or I→T.Conclusion Norovirus GZ20133135 belonged to the G Ⅱ-4 Sydney2012 variant.In This study,GZ20133135 full sequence information can be used not only as a full-length NoV variant sequence standard for future comparison studies,but also as useful material for the public health by enabling the diagnosis,vaccine development,and prediction of new emerging variants.

Objective To understand the molecular epidemiology of norovirus from 5 outbreaks of gastroenteritis during Dec.2012 to Jan.2013 in Guangzhou.Methods Epidemiologic data and specimens were collected from 5 gastroenteritis outbreaks in Guangzhou.274 specimens were detected for norovirus by RT-PCR methods and PCR products were sequenced.Sequence alignment and phylogenetic analysis were performed by using Clustal X 1.83 and MEGA 5.05 programs.Statistical analysis was performed by using SPSS 17.0 program.Results The total positive rate of norovirus was 20.07％ (55/274).The positive rate was 8.70％ (2/23) in University A,36.36％ (8/22) in Kindergarten B,36.07％ (22/61) in University C,100％ (5/5) in community health service center D,11.04％ (18/163) in University E.The positive rate was 100％ (6/6) among age group ≥ 60 years old,47.37％ (9/19) among 10-19 age group and 36.36％ (8/22) among age group ＜ 10 years old.Phylogenetic analysis showed that 33 samples were infected by the new variant-Sydney 2012 of norovirus GⅡ.4.Conclusion Norovirus was one of the main pathogens causing acute gastroenteritis outbreaks and GⅡ.4 Sydney 2012 variant was identified as the predominant strain in Guangzhou.

Objective:To investigate the pathogenic spectrum of hand, foot and mouth disease(HFMD)from 2017 to 2019 in Guangzhou, and analyze molecular epidemiological characteristics of coxsackievirus A6 (CV-A6).Methods:Enterovirus A71 (EV-A71), CV-A16, CV-A6 and enterovirus were tested by reverse transcription-quantitative polymerase chain reaction(RT-qPCR). The CV-A6 representative samples were isolated and the VP1 region of isolates were amplified and analized by Mega5.0 and SeqMen.Results:A total of 7 578 enterovirus-positive specimens were detected from 2017 to 2019, 320 specimens were positive for EV-A71, 1481 specimens were positive for CV-A16, 3171specimens were positive for CV-A6, and 2606 specimens were positive for other enterovirus. Children under the age of 5 years were the most vulnerable population, and the male/female incidence ratio was 1.56∶ 1.The incidence occurred in all seasons, one peak between May and July, the other between September and November. The virus was isolated from 80 CV-A6 positive specimens and the full length of VP1 gene region was sequenced and nucleotide sequence similarity analysis was performed. The nucleotide homology in the VP1 region was 93%-100%, and the amino acid homology was 98%-100%. The nucleotide homology with the CV-A6 prototype strain (Gdula) VP1 region was 79%-81%, and the amino acid homology was 95%-97%. The nucleotide homology with the representative strain of D3 subtype was 92%-98%, and the amino acid homology was 98%-100%. Phylogenetic tree shows that all CV-A6 belonged to the sub-genotype D3 and distributed in multiple branches.Conclusions:CV-A6 is emerging as one of the major pathogen causing HFMD in Guangzhou, and all insolates belonged to D3 subtype. Closely monitoring the molecular characteristics of CV-A6 and changes in the pathogen spectrum can provide scientific basis for HFMD prevention and control.

Objective:To explore the potent effects of denatonium benzoate(DB)on Mas-related G protein-coupled receptor-B2(MrgprB2)-mediated rat mast cell degranulation.Methods:RBL-2H3 cells were treated with DB overnight,before challenged with MrgprB2 ligands substance P(SP).The release of β-hex from MrgprB2-activated RBL-2H3 was detected by substrate method.Detec-tion of LTC4,IL-6,TNF-α and cPLA2 activity were performed by ELISA.The Ca2+influx and the expression of RBL-2H3 MrgprB2 re-ceptors were measured by fluorescence assay.Results:The results showed 10 μmol/L,50 μmol/L,80 μmol/L,100 μmol/L DB treat-ments promoted β-hex and LTC4 releases from activated RBL-2H3,accompanied by increased Ca2+mobilization and cPLA2 activa-tion.DB treatments did not affect IL-6 and TNF-α LTC4 releases in MrgprB2-activated RBL-2H3,as well as the levels of MrgprB2 ex-pression in mast cells.Conclusion:Taken together,DB enhanced the MrgprB2-mediated RBL-2H3 degranulation in vitro,probably by up-regulating Ca2+mobilization in activated cells.