【Objective】In order to establish the foundation for transgenic mouse model,the human thalassemic gene(β654) was cloned and sequenced.【Methods】The human β654 gene was amplified by PCR,and cloned into the plasmid BGT51 in which the human β gene was cut out aforehand.The recombinant plasmid was certified by enzyme-digestion,reverse dot hybridization and sequencing.【Results】A recombinant plasmid was obtained,which contained the human β654 gene in the correct recombinant direction.Sequencing showed that the cloned insert was correct.【Conclusions】The recombinant plasmid constructed is useful for establishing a transgenic mouse.

【Objective】 To investigate the incidence of β-th alassemia (β-thal) heterozygote carrying α-thalassemia (α-thal ) 1 gene in Guangdong area. 【Methods】 β-thal genes were screened by reve rse dot blotting (RDB). In β-thal DNA samples α-thal 1 genes were am plified using gap-PCR method. 【Results】43 α-thal-1 cases were identifi ed among the 500 β-thal traits. The rate is 8.6%. 【Conclusion】 In Guangd ong area the incidence of β-thal heterozygote carrying α-thal-1 gen e is 8.6%, which should be paid much attention in the genetic counselling and pr enatal diagnosis.

AIM: To clone human ?-globin gene carrying a thalassemic mutation IVS II654(C→T) and establish a eukaryotic expression system for high-level expression of human ? IVS II654 gene in mouse erythroleukaemia(MEL) cells. METHODS: The fragments of human ? 654 gene isolated from the ? thalassemia patients homozygous for the ? 654 mutation were amplified by PCR, and cloned to plasmid pBGT51. Then, the human ? LCR and ? 654 gene were subcloned from plasmid pBGT51 to the stable mammalian expression vector pcDNA3.1+ together, and the MEL cells were transfected with this vector using commercially available cationic lipid FuGENE6. The MEL cells were induced for further maturation by DMSO and the expression of human ? 654 gene in the MEL cells was identified by RT-PCR. RESULTS: A mammalian expression system of human ? thalassemic mutation ?IVS II654(C→T) was established. CONCLUSION: The level and the reliability of expression of human ? 654 gene in the MEL cells in vitro are similar to that in vivo in human body. This may be a valuable gene therapy model for human ? thalassemic mutation ?IVS II654(C→T).

AIM: To investigate whether celecoxib induces gastric cancer cell apoptosis in a COX-2 non-expression cell line.METHODS: The COX-2 protein was examined by western blotting.Fluorescence microscopy,DNA agarose gel electrophoresis and flow cytometry analysis were used to test apoptosis.RESULTS: COX-2 was expressed in AGS but not MGC-803 gastric cancer cell line;Selective COX-2 inhibitor celecoxib induced MGC-803 cell line apoptosis in a concentration and time-dependent manner.CONCLUSION: Celecoxib induces apoptosis in COX-2 non-expression gastric cancer cells.

Objective To explore the value of three antibodies in the differential diagnosis of papillary thyroid carcinoma ,by de‐tecting the expression of HBME‐1 ,CK19 and CD117 in papillary thyroid cacinoma ,thyroid follicular adenoma and Hashimoto′s thy‐roiditis tissues .Methods Totally 85 cases were collected from January 2013 to December 2015 ,including papillary thyroid cacino‐ma ,thyroid follicular adenoma and Hashimoto′s thyroiditis .They were immunohistochemical stained by HBME‐1 ,CK19 and CD117 .SPSS16 .0 software was used to analyze the relationship between the staining results with different pathological changes . Results The positive rates of HBME‐1 ,CK19 and CD117 were 87 .3% ,98 .2% ,and 7 .3% ,respectively .The positive expression of them in benign and malignant groups had significant difference (P< 0 .05) and their consistency checking Kappa were 0 .582 , 0 .551 ,and 0 .874 ,respectively .Conclusion In the differential diagnosis of papillary thyroid carcinoma and benign lesions ,CD117 is better than HBM E‐1 and CK19 .It′s possible to use a combination of them in practice .

Objective To identify the genotypos of extended spectrum β-1actamase (ESBLs)-producing of Escherichia coli ( E. coli) clinical strains isolated from the Macao and compare the results with the genotypes of clinical strains collected in the first Clinical College, China Medical University (CMU) in Shengyang. Methods The clinically isolated E. coli strains including 209 strains from Macao and 150 strains from CMU were collected. Based on the standard of CLSI2006, the ESBLs-producing strains was identified and its isoelectric point(pI) value was detected by isoelectric focusing (IEF) method. The pI values were used to design the primers for PCR amplification. The amplified DNA sequences were then compared with the GenBank and the ESBL genotypes were confirmed. Results ( 1 ) The positive rate of ESBLs-producing strains of E. coli was 30. 1% (62/209) from Macao and 54. 0% (81/150)from CMU. (2)The genotype of 56 (90. 5% ) β-lactamase(ESBLs)-producing E. cull strains from Macao was CTX-M56. Most of them were CTX-M-14 (76. 2% ), other genotypes including CTX-M-9 (4. 8% ), CTX-M-22 (3.2%), CTX-M-24(3.2%), CTX-M-27(1. 6% ), and CTX-M-15( 1.6% ) were found. Six strains were unidentified. (3)The genotype of 74(91.5% )β-lactamase(ESBLs) -producing E. coli strains from Shenyang was CTX-M. Most of them were CTX-M-14 (65.4%), other genotypes including CTX-M-3 ( 13.6% ), CTX-M-24 (4. 9% ),CTX-M-22(2.5%), CTX-M-15(2.5%), CTX-M-9(1.2%) and CTX-M-28(1.2%) were found. Seven strains were unidentified. Conclusions CTX-M genotypo was the mostly identified ESBL-preducing E. Coli strains from Macao and the results were similar with that from CMU. Among them, the CTX-M-14 was the major genotype. Other genotypes included CTX-M-9, CTX-M-15, CTX-M-22, CTX-M-27, and CTX-M-24.However, two genotypes of CTX-M-3 and CTX-M-28 were not found in the clinical isolates in Macao and one genotype of CTX-M-27 was not found from the CMU clinical isolates.

OBJECTIVETo select the premium alert threshold for major communicable disease by using the control graph alert technique based on the local disease information.

METHOD8 major communicable diseases in Songjiang district were ascertained by analysis of the national early warning detection information system which include the other diarrhea, mumps, chickenpox, scarlet fever, rubella, hand foot mouth disease, influenza and dysentery; weekly reported cases from 2008 to 2011 were used to establish the early detection model (PERCENTILE (array, x), array (4×5), x = 0.05, 0.10…0.95) by moving percentile method, next applying the established early detection model and the golden standard (AKX(-)D ± 2s) to predict the expected weekly cases in 2012 respectively, and then ascertain the predict results by comparison with the actual weekly cases in 2012 respectively, finally the premium threshold was selected by comparison of the model predicted results with the golden standard predicted results after comprehensive consideration of the sensitivity, specificity, positive predictive value and negative predictive value and receiver operating characteristic (ROC) curve.

RESULTSThe premium alert threshold for mumps, other diarrhea and rubella was P90, dysentery was P75, scarlet fever and chickenpox was P80, and the premium threshold for hand-foot-mouth disease (HFMD) and influenza was P95, the sensitivity of 8 major communicable diseases were 100%, 100%, 86%, 100%, 100%, 100%, 94%, 100%, respectively; the specificities were 92%, 73%, 72%, 77%, 73%, 92%, 66%, 80%, respectively; the positive predictive values were 43%, 40%, 32%, 8%, 24%, 20%, 59%, 47%, respectively; and the negative predictive values were 100%, 100%, 97%, 100%, 100%, 100%, 96%, 100%, respectively. The national recommended alert thresholds for the 8 major communicable diseases were P80, except for chickenpox (P50) and HFMD (CUSUM).

CONCLUSION6 out of 8 major communicable diseases' early detection thresholds in Songjiang district should be adjusted according to the analysis results. Premium alert threshold selection need to consider the local disease report and the characteristics of infectious diseases to upgrade the early detection capability.

China ; epidemiology ; Communicable Disease Control ; methods ; Communicable Diseases ; epidemiology ; Disease Notification ; Disease Outbreaks ; prevention & control ; Humans ; Models, Theoretical ; Population Surveillance ; methods