Laboratory diagnostics is one of the most fast developing medical sciences.But the teaching quality falls behind due to the traditional education model.Based on the concept of ‘three-stage fusion' of the subject and ‘five abilities' of the training objects,‘three-highlight & stereo' teaching style was established and put through the course,which means to emphasize function,practice,effect in optimizing teaching content and methods,and construct stereo platform including information system,online course,laboratory equipments,research activities and teacher training program.The results showed a significant improvement of the students' knowledge mastering and utilizing capability.And the teaching situation was well re-created.Furthermore,the teaching team was much more powerful than ever.It suggests that ‘three-highlight and stereo' teaching style is a successful explore and practice of teaching reform of laboratory diagnostics on adapting for internationalization of education tendency.

0bjective To investigate the serum total bilirubin level of healthy population in northwest area of China.Methods According to the guiding principle and the screening standard of C28-P3,722 individuals conforming to the screening standard from October 2011 to January 2012 were collected.Serum total bilirubin was determined with diazo reagent method(Roche)on the P module of Hitachi 7600 automatic biochemistry analyzer,and the vanadate reagent method(WAKO)on the D module of the Hitachi 7600 automatic biochemistry analyzer,respectively.The detection results were analyzed between different analysis systems,between country and city and among different age groups by SPSS13.0.The top and bottom limitations of 95% reference interval recommen-ded by the C28-P3 file were calculated by the non-parametric method.Results Serum total bilirubin detection results had no statis-tically significant differences between the two kinds of detection methods,between rural and urban populations,among all ages and genders (P >0.05).The ultimate reference interval of serum total bilirubin in the northwest area was 2.19 -29.29 μmol/L.Con-clusion The differences of serum total bilirubin reference interval exist between the healthy population in the Northwest area of China and the current reference interval used in domestic and foreign population.Establishing the reference intervals of new bio-chemical test item suitable for China′s population will provide the scientific basis for the evaluation of disease diagnosis,treatment, prognosis judgment and health assessment in the Chinese population.

Objective To study the reference interval of serum iron and magnesium of healthy people in northwest region . Methods 722 healthy people aged 18- <80 were recruited according to the screening criteria of enzymatic multi-center study of International Federation of Clinical Chemistry (IFCC) .Atomic absorption spectrophotometry and Roche assay kit were employed for serum iron ,magnesium detection .Results Differences of serum iron ,magnesium detection results obtained by two different methods ,between urban and rural people and among different age periods showed no statistically significant (P>0 .05) ,and the re-sults was combined into one group .Difference of iron test results between male and female was statistically significant (P<0 .05) , and grouping was performed according to gender ,that was ,male:10 .05-36 .21 mmol/L ,female:6 .19-30 .87 mmol/L .Difference of magnesium test results between male and female showed no statistical significance (P<0 .05) ,and was combined into one group , with the reference interval of 0 .74-1 .06 mmol/L .Conclusion The iron and magnesium 95% reference intervals of healthy people in northwest region are narrower than those of the National test results .

Objective: To clone human carboxypeptidase A1 and its active center gene as well as construct recombinant vector for expression before analysis their activities. Methods: CPA1 and CPAlactive center gene were amplified by RT-PCR from pancreas tissue, and then sequencing was carried out. The correct target genes were cloned into prokaryotic vector pGEX-4T-1 and transformed into E. coli BL21 before sequence analysis. After induced by IPTG, gene products were analyzed by SDS-PAGE. Target expressed proteins were denaturated, renaturated, purified and evaluated through MTT and agar colony form test. Results: Human carboxypeptidase Al and its active center gene were cloned successfully. New expected protein band of Mr 66,000 and 46,000 appeared on SDS-PAGE after inducement. Both of the expressed proteins have catalytic activity in vitro, but the activity of the latter is inferior when applied to tumor cells. Conclusions: Human carboxypeptidase A1 and its active center gene were cloned successfully. Their prokaryotic expression products were obtained too. The expressed proteins have catalytic activity in. vitro. A new prosperous beginning of further improvement for CPA1 therapy system has been established based on CPA1 and its active center gene in terms of ADEPT against prostate cancer to clinical application.

Objective To investigate the application and significance of the MS/MS method and the HPLC method for the screening and diagnosis of genetic metabolic disease phenylketonuria (PKU) .Methods The MS/MS method and the HPLC method were adopted to analyze the concentrations of phenylalanine(Phe) and tyrosine(Tyr) and its ratio in the dried blood spot specimen on filter paper and the whole blood specimen in 1 860 children aged from 3 d to 11 years old .Results The linear ranges of Phe by the MS/MS method and the HPLC method were 26 .02 - 101 .11 μmol/L and 32 .04 - 132 .50 μmol/L ,which of Tyr were 41 .50 -253 .07 μmol/L and 32 .85 - 111 .50 μmol/L ,the average recovery rates of Phe were 97 .36% and 98 .43% ,which of Tyr were 96 .71% and 98 .99% ,in‐run CV of Phe were 4 .31% and 3 .97% ,which of Tyr were 4 .09% and 4 .01% ,between‐run CV of Phe were 5 .73% and 4 .58% ,which of Tyr were 6 .01% and 5 .24% ,respectively .Conclusion Both methods can sensitively and specif‐ically detect blood Phe and Tyr concentrations and meet the requirements of screening and diagnosis of PKU .

Objective To evaluate the strength of tobacco control at or below county-level governments of Zhejiang Province,to explore the strategies and measures on tobacco control,and promote a smoke-free government.Methods By multistage stratified random sampling method,12 government ageucies were selected in 4 counties.The information of tobacco control strength and smoke-free environment creation was collected by interview and field observation,and the scores were calculated by using the percentage assignment method.An intercept suuvey was carried out in four counties and there were 405 civil servants under investigation.The data were analyzed by frequency statistic and correlation analysis.Results The smoking rate of civil servants was 17.8％,and the secondhand smoking rate was 61.9％;84.7％ of civil servants adnmitted the harm of smoking and 96.8％ admitted the harm of secondhand smoking.Support for forbidding the cigarette advertisements was highest and increasing the cigarette price was the weakest.The scores of smoke-free environment creation of project counties showed that the average score was 50.60,Zhuji and Luqiao got a maximum score of 57.14,Haiyan got a minimum score of 42.86.The scores of tobacco control strength showed that the average score was 82.35,Zhuji got a maximum score of 92.16,followed by Luqiao,Jiande and Haiyan counties,82.35,80.39 and 74.51 respectively.The scores of tobacco control strength,include policy,political will,capacity,and tobacco control efforts were the highest in Zhuji county.The results of correlation analysis showed that the total score of tobacco control strength had significant association with the score of smoke-free environment creation,smoking rate of civil servants,and approval rate of anti smoking in public places (P＜0.05).Conclusion The overall situation of tobacco control strength at or below county-level governments of Zhejiang province is good,but there is a relative shortage of smoke-free environment creation.Civil servants master the knowledge related to tobacco and support tobacco control policy.We should further promote the construction of smoke-free government.More effective smoking control measures should be conducted in order to raise the strength of tobacco control and finally implement the tobacco control legislation.

Objective To understand the prevalence of tobacco use among college students in different populations by sex,age and major,and to provide evidence for smoking control and health decision.Methods A total of 4 638 college students selected in 21 colleges through stratified multi-stage cluster sampling from Zhejiang Province were surveyed by using questionnaire analysis.Indicators such as tried smoking,current smoking,and secondhand smoke exposure,etc.,were calculated by weight on age proportions from the numbers of college students in Zhejiang.Results The prevalence of tried smoking and currcut smoking among college students were 29.72％ and 10.57％,respectively.The prevalence of tried smoking was 46.52％ for men,18.25％ for women;28.57％ for undergraduate,31.53％ for junior college;the prevalence of current smoking was 18.86％ for men,3.86％ for women;8.88％ for undergraduate,13.23％ for junior college students.The snoking rate in college students increased with age,the group of ≥21 Years was the highest (37.87％,16.09％).The lowest level of smoking was medical student (21.76％,2.19％).The prevalence of secondhand smoke exposure in outdoor and indoor places,and dormitory among students were 65.49％,60.33％,and 23.66％,respectively.In the past 30 days,college students have seen tobacco advertisement (47.38％),smoking scenes (60.20％),and got tobacco related gifts (10.40％),free tobacco products (8.93％);college students have seen information about tobacco control (60.50％),cigarette pack warning labels (58.45％),and learned the tobacco knowledge in class in the past 12 months (14.03％).Conclusion Current prevalence of smoking in college students in Zhejiang province remained at a high level,the rate of secondhand smoke exposure is high and in college students there was lack of tobacco control education in class.The results indicated that the policy about tobacco control in school was not enough and the task of health promotion should be taken more arduously.

Objective To explore the determinants of willingness to quit smoking among pre-pregnancy male smokers, and to provide evidence for smoking cessation strategies. Methods An intercept survey was carried out in four cities in Zhejiang, and there were 552 pre-pregnancy male smokers from 1401 newly wedded couples under investigation. Result The pre-pregnancy male smoking rate was 39.82%, and only 6.88%of pre-pregnancy male smokers were categorized as highly nicotine-dependent. The rate of recent willingness to quit smoking was 50.54%, and reasons for quitting were as follows:a planned pregnancy (62.50%), an example for children (47.70%), and objections from family members (45.22%). The tobacco knowledge among pre-pregnancy male smokers is relatively poor, only 49.64% of smokers were aware 8 points in 11 questions; 88.75% of smoker's wives supported their husband to quit smoking. The associated factors of smoking cessation include: tobacco knowledge (OR=0.38), health self-assessment well (OR=1.90), wife's willing for husband to quit smoking (OR=2.87). Conclusion Compared to ordinary people, the proportion of intention to quit smoking was higher in pre-pregnancy male smokers. To strengthen health education and make wife initiating in husband's quitting programs would be more effective.

Objective To construct fusion gene and prokaryofie expression plasmid encoding the Rv1884c and Rv0867c genes in Mycobacterium tuberculosis (M.tb).The fusion protein wsg expressed efficienfly in E.coli cells.Methods The Rv1884c and Rv0867c genes were amplified by polymerase chain reactions (PCR) with specific primers from genomic DNA of M.tb H37Rv strain,and cloned into pGEX-4Tland pUC19 vectors.Rv1884c and Rv0867c were subcloned into the expression vector pGEX-4T-1 and pPRO-EXHT followed by DNA sequencing.The plasmids were transformed into E.coli DH5α and induced to produce GST-fused Rv1884c and His-fused Rv0867c fusion protein.The protein molecular weight and expression format was analyzed by SDS-PAGE.Results The recombinant expressive vectors pGEX-4T-1Rv1884c and pPRO-EXHT-Rv0867c were constructed.The DH5α strains of E.coli with recombinant plasmid showed high level of Rv1884c and Rv0867c gene expressions after IPTG induction.The SDS-PAGE showed that the plasmids expressed Rv1884c and Rv0867c fusion proteins with molecule weight of 45 000 and 80 000.The recombinant protein accounted for 18.3%and 23.7%of total bacteria protein.The expressed proteins could be purified via GSTrao FF and Ni2+-NTA system kits in denatured condition.Conclusions Mycobacterium tuberculosis Rv1884e and Rv0867c genes have been cloned and expressed Successfully in E.coli DH5α.The results lay a basis for further study of fast culfivation in Mycobacterium tuberculosis and investigation of their activities and functions.

Objective:To construct a short hairpin RNA(shRNA) eukaryotic expression vector of the survivin gene,and investigate its inhibitory effect on the survivin expression in human cervical cancer cells.Methods:We designed and synthesized 2 pairs of survivin-specific small interfering RNA primers(s1 and s2),cloned them into the eukaryotic expression vector pSilencer 2.1-U6 neo by DNA recombined techniques after annealing connection reaction,and transfected them respectively into human cervical cancer cell line HeLa using LipofectAMINE2000 after identificationby restrict endonuclease digestion and DNA sequencing.Then we selected the positive clones by G418 and detected the expression of survivin mRNA by semi-quantitative RT-PCR and that of the survivin protein by Western blot.Results:The survivin shRNA eukaryotic expression vectors pSilencer2.1-s1 and pSilencer2.1-s2 were successfully constructed.Positive clones were obtained by screening with G418 for 24 days,the survivin expression in HeLa cells decreased in different degrees after transfected with pSilencer2.1-s1 and pSilencer2.1-s2,and the latter showed a better interference efficiency.Conclusion:The shRNA eukaryotic expression vector of the survivin gene we constructed,with its improved interfering efficiency,has paved the way for further research on its role in regulating the biological behavior of cervical cancer cells.