Objective To construct the prokaryotic expression vector of human procalcitonin(PCT) and obtain the highly purified recombinant PCT protein.Methods PCT primers were designed based on the sequence of PCT gene from NCBI,and PCT gene was amplified by PCR.Then,the recombinant vector PCT/pET-22b(+) was constructed and transferred into E.coli BL21 to induce the expression of recombinant PCT protein.Last,the recombinant PCT protein was purified by the nickel column affinity chromatography and identified by Western blot and the colloidal gold method,respectively.Results The results of agarose gel electrophoresis showed that the product of PCR amplification was about 350 bp.The homology comparison analysis revealed that the PCT gene fragment(348 bp) was successfully inserted into pTG19-T vector,and that no any base mutation was found.When the recombinant plasmid of PCT/pET-22b(+) was digested with BamH Ⅰ and HindⅢ,two pieces of about 350 bp and 5 500 bp were obtained.SDS-PAGE showed that the recombinant PCT protein with about 14 000 of Mr was existed in soluble form,and was easily obtained by the nickel column affinity chromatography.Moreover,western blot and the colloidal gold method demonstrated that the recombinant PCT protein was successfully expressed and contained histidine label(His-tag).Conclusion The recombinant vector PCT/pET-22b(+) is successfully constructed by the recombinant DNA technology,and the recombinant PCT fusion protein is successfully obtained by the nickel column affinity chromatography.

Objective To clone and express the thioredoxin(Trx)from RH strain tachyzoites of Toxoplasma gondii,estab?lish the prokaryotic expression vector and purify the recombinant protein,then produce the polyclonal anti?Trx antibody in rab?bits. Methods Trx fragment was amplified by PCR and cloned into the pET?28a(+)vector,and the recombinant protein was in?duced with IPTG and purified by Ni?NTA affinity chromatography. The polyclonal antibody specificity was detected by Western blotting. Results The trx gene was amplified from T. gondii cDNA by PCR. The recombinant plasmid trx/pET?28a(+)was use?fully constructed,and the recombinant TRX protein was expressed and purified. The TRX polyclonal antibody was also ob?tained. The specific band of TRX was detected by Western blotting. Conclusion Western blotting can detect the specificity of polyclonal anti?Trx antibody,which will facilitate the biological functions of Trx.

Objective: To establish a method for rapid determination of 47 volatile organic compounds in the air of workplace using portable gas chromatography-mass spectrometer(GC-MS). Methods: The mixed standard gas with different concentration levels was made by using the static gas distribution method with the high purity nitrogen as dilution gas. The samples were injected into the GC-MS by a hand-held probe. Retention time and characteristic ion were used for qualitative analysis,and the internal standard method was usd for quantitation. Results: The 47 poisonous substances were separated and determined well. The linear range of this method was 0.2-16.0 mg/m³,and the relative standard deviation of 45 volatile ovganic compounds was 3.8%-15.8%. The average recovery was 79.3%-119.0%. Conclusion The method is simple,accurate,sensitive,has good separation effect,short analysis period, can be used for qualitative and quantitative analysis of volatile organic compounds in the workplace, and also supports the rapid identification and detection of occupational hazards.

This study was aimed to evaluate the clinical effect and safety of Qing-Fei Tong-Luo (QFTL) ointment for treating children with pneumonia.Randomized controlled trial (RCT) was conducted among 460 cases of children with pneumonia.The observation group was given QFTL ointment combined with basic treatment.And the control group was only treated by basic treatment.Evaluation was given on the total clinical efficacy,disappeared time of fever,cough,expectoration,shortness of breath,and medication safety.The incidence of respiratory diseases was followed up on the 30th days after drug withdrawal.The results showed that in the aspect of clinical efficacy between two groups,the cure rate of the observation group was 98.26％,and that of the control group was 93.89％,with statistic significance (P ＜ 0.05).The cure rate of the observation group was better than that of the control group.There was statistical difference on expectoration disappeared time (P ＜ 0.05).There was no statistical difference on disappeared time of fever,cough and shortness of breath (P ＞ 0.05).There was statistical difference on the incidence of respiratory diseases on the 30th days followed-up after drug withdrawal (P ＜ 0.05).There was no statistical difference on the incidence of upper respiratory tract infection,pneumonia and asthma (P ＞ 0.05).No adverse reactions occurred in the observation group.It was concluded that QFTL ointment combined with basic therapy on the treatment of pneumonia in children was significantly better than the control group in the aspect of clinical efficacy,expectoration disappeared time and the incidence of bronchitis.It is safe and effective.The prognosis is good and worthy of promotion in the clinical practice.