We investigated the effects of γ-interferon and exogenous indole on the growth of domestic dominant standard strains and clinical straìns of Chlamydia trachomatis E-UW-5/Cx,and compared with the dominant strains of D-UW-5/Cx abroad.We used DMEM-10,DMEM-10 containing 5 ng/mL recombinant human interferon gamma (referred to as DMEM-10+IFN) and DMEM-10 containing 5 ng/mL recombinant human interferon gamma and 50 μM exogenous indole (referred to as DMEM-10+IFN+IND) to culture C.trachomatis,and then we fixed it with methanol to count inclusions after 48 hours,observing the influence of r-interferon and exogenous indole on the growth of C.trachomatis standard strains(E,D) and clinical strains.Results showed that the count of Chlamydia inclusion bodies in DMEM-10+IFN group was significantly lower than others (P＜0.05);no significant difference was found (P＞0.05) between the count of DMEM-10 group between DMEM-10+IFN+IND group.There were no significant difference between the E and D standard or clinical strains (P＞0.05).Under the effect of IFN-γ,the growth of domestic dominant strain E-UW-5/Cx C.trachomatis was significantly inhibited.After adding exogenous indole,C.trachomatis can escape the scavenging activity of IFN-γγto restore the infection vitality.

The aim of the present study was to work on the efficiency of differential diagnosis of native hapten-gel diffusion assay (NH-GD) on the background of vaccination with S2 or Rev.1.The conditions of NH-GD assay was firstly optimized,its sensitivity,specificity,repeatability and ability of differential diagnosis were determined respectively,and its test result was compared with that of fluorescence polarization assay (FPA).The results showed NH-GD assay with good specificity and repeatability could differentiate Brucella-infected antibody from vaccinated antibody after vaccination with S2 or 122 days after vaccination with Rev.1.And the result of NH-GD assay was highly consistent with that of FPA,which was simple to operate and needed a few simple equipment.Therefore,NH-GD assay was a good method for sheep brucellosis surveillance in China and especially suitable for application in grass-roots areas.

To clone and express 27 kDa cysteine protease (CP) gene of Spirometra erinacei plerocercoid,and analyze the biology characteristics,a total of RNA was extracted from the plerocercoids and reversely transcribed into cDNA.The 27kDaCP gene was amplified by PCR and cloned into pM-19T vector for sequencing.The accurate sequence was subcloned into the expression vector pET-28a (+).The recombinant plasmid was transformed into Transetta (DE3) and the expression protein induced by IPTG.The recombinant protein was purified by Ni2 + affinity chromatography,and analyzed by SDS-PAGE and Western blotting.The 27 kDa CP gene and its expression protein were predicted and analyzed by bioinformatics analysis tools such as NCBI and ExPASy.Results showed that the ORF length of 27 kDa CP gene sequence was 1 011 bp,and the removed signal peptide sequence was 954 bp with the submission number of ANA52569 in GenBank.The whole sequence of 27 kDa CP (Mr 35 669.9,pI 5.92) was 317 amino acids conferred from cDNA,which belongs to the Peptidase_C39_like superfamily.The pET-28a (+)-27kDa-CP was expressed under the induction of IPTG.Western blotting analysis showed that the purified protein reach expectancy,and had better response with positive serum of Spirometra erinacei plerocercoid infection.In conclusion,the 27 kDa CP gene of Spirometra erinacei plerocercoid is successfully cloned and expressed and knowing coded sequences and bioinformatic.

To analyze the genotype of Echinococcus granulosus found in Liaoning Province,hydatid cyst was taken from the patient's liver,DNA of cyst was isolated and mitochondrial cox1 gene was sequenced substantially.Sequence results was analyzed by Blast and compared with similar strains in GenBank.Results showed that molecular analysis confirmed the echinococcosis by identical sequence of our strain with E.granulosus (AF297617) G1 genotype,with 0.1％ difference.And the sequence of our strain was consistent with 25 other strains in different countries.In conclusion,genotyping of mitochondrial gene coxl is a very useful tool for diagnosis of echinococcosis.We first reported a strain of E.granulosus in Liaoning Province,which provide novel molecular epidemiological data of this zoonosis.

Chlamydia psittaci is a causative agent of psittacosis,which can infect a wide range of hosts including birds and humans.However,information regarding C.psittaci infection in pigeons is scarce.In the present study,a total of 399 fecal samples from pigeons were collected from Jilin Province,northeastern China,between March and May 2015,and examined by nested PCR amplification of outer membrane protein A (ompA) gene.The overall Chlamydiosis prevalence was 5.01％ (21/399),with 3.19％ in Changchun City and 9.40％ in Jilin City.Furthermore,breed was the major risk factor associated with Chlamydia infection in pigeon,boiler pigeons had a prevalence of 7.49％,whereas no C.psittaci was detected in racing pigeons.Sequence analysis of the ompA gene revealed that all the identified isolates represented C.psittaci genotype B.Our results firstly indicated the presence of zoonotic C.psittaci in boiler pigeons in Jilin Province,northeastern China,and effective measures should be implemented to reduce the risk of C.psittaci transmission from pigeons to humans.

We investigated the effects of IFN-γ on Chlamydia psittaci (Cps) infection.HeLa cells were treated with different concentrations of recombinant human IFN-γ (5 ng/mL,25 ng/mL,50 ng/mL) after infecting with C.psittaci 6BC,then the number and morphology of C.psittaci inclusion bodies were examined after 48 hours.C57BL/6J mice were intranasally infected with 2 × 106 IFUs C.psittaci 6BC,and intraperitoneally administrated with 10 μg recombinant murine interferon-γ 24 hours prior or post infection,then body weight,activity and survival rate were recorded.The histopathology of mice livers and lungs was analyzed by HE staining on day 5 or day10 post infection.And the chlamydial inclusion bodies were titrated in the lung homogenates of mice sacrificed on day 5 after infection.The inclusion body numbers of recombinant human IFN-γ treated groups (by 5ng/mL,25ng/mL,50ng/mL) were significantly less than that in the control group (23.8±5.1)× 106,(10± 3.58) × 106,(8.0±2.22) × 106,(43.3±11.05)× 106,respectively).And the morphology of inclusion bodies in IFN-γ treated HeLa cells was irregular and much smaller.We also found that IFN-γ could significantly improve the survival rate,reduce acute clinical manifestations and pathological injurery of lung and liver in C.psittaci respiratory tract infected mice model.So we summarized that IFN-γ can mediate strong immunological protection during acute C.psittaci early infection.

Severe infectious diseases,i.e.antibody-dependent enhancement (ADE) resulted from successive infection with different serotypes of dengue virus.After its introduction into Brazil in 2015,Zika virus has spread rapidly to more than 60 countries and regions by the end of November 2016.Some south-east Asian countries including China have also reported cases of ZIKV infection.In recent studies,it was observed that sera cross-reactivity antibodies or such monoclonal antibodies have been elicited by two domains,ED1 and ED2,of envelope (E) protein on Zika or/and Degue virus,and ADE was easily induced by such antibodies.Dengue fever epidemic often occurred in Chinese coastal provinces each year.Then,it will be followed by Zika virus disease.Therefore,we must pay attention to and propose replying measurement for it.

Currently,the BCG is used to prevent tuberculosis,but the immune effect is not ideal due to varied reasons.The existence of drug-resistant strains of tuberculosis and the increased prevalence and incidence of AIDS have leaded to the increased incidence of TB year by year.Therefore,the development of new tuberculosis vaccine is imminent In this paper,the latest research results in recent years for tuberculosis live vector vaccines were summarized,which provide a theoretical reference for further research and development of new TB vaccines.

We evaluated clinical application effect of gene chip for detection of rifampin (RFP) and isoniazid (INH) resistance in Mycobacterium tuberculosis (MTB).Rifampin and isoniazid drug-resistance gene loci were detected by gene chip with sputum specimens from smear-positive tuberculosis patients and clinical strains,comparing the results of detection.BACTEC MGIT 960 drug susceptibility test results were used as control to evaluate the detection performance of gene chip.The sequences of the polymerase chain reaction products of the rpoB,katG and inhA genes from 999 strains identified as Mycobacterium tuberculosis were determined to confirm the mutations by DNA sequencing.Results showed that 100 cases were identified as nontuberculous mycobacteria by gene chip in the 1 108 cases of smear-positive samples.Among the rest 1 008 samples,there were only 9 cases of microarray results different from BACTEC MGIT960 culture-positive strains,achieving the coincidences of 99.1％.Compared with BACTEC MGIT 960 drug susceptibility test results,the gene chip method displayed a concordance of 98.1 ％ and 94.5 ％ for RFP and INH respectively in the 999 strains.Compared with the DNA sequencing method,the accuracy of gene chip method was 99.6％ for rifampin resistance and 99.8％ for isoniazid resistance.It's concluded that the gene chip technology can quickly and accurately detect rifampin and isoniazid resistance in MTB and can be used directly for the detection of sputum samples.

We investigated the correlation between toxin gene exoS,exoU and fluoroquinolone resistance in lower respiratory tract infection with P.aeruginosa so as to provide guidance for reasonable treatment of clinical infections.We collected P.aeruginosa of sputum samples in hospitalized patients from October 2015 to March 2016.The antimicrobial susceptibility was tested by liquid dilution method.The exoS and exoU genes were detected by PCR technique.Results showed that forty-six P.aeruginosa strains were identified from sputum.The exoS and exoU gene positive rate were 86.96 ％ (40/46) and 69.57 ％ (32/ 46) respectively,and the highest proportion of genotype was exoS+/exoU+ (60.87％,28/46).Among them,36.96％ (17/ 46) were multiple drug-resistant bacteria(MDR).Fluoroquinolone non-sensitive (FQ-NS) strain were 78.95％ (15/19) for MDR and 89.47 ％ (17/19) exoU gene were positive,which was significantly higher than the fluoroquinolone sensitive strains (FQ-S).Compared with the FQ-S strain,FQ-NS strains were serious drug resistance.The drug resistant rate of eefepime and aztreonam were more than 70％,and then meropenem and imipenem were more than 50％.The drugs of lower resistance rate in FQ-NS strain had polymyxin B(10.53％,2/19),amikacin(10.53％,2/19),ceftazidime (15.79％,3/19) and gentamicin (21.05％,4/19).P.aeruginosa of lower respiratory infection carried toxin genes exoS and exoU were higher,the main genetpy was exoS+/exoU+.FQ-NS strains were higher drug resistance rate and a higher proportion of exoU+ strains than FQ-S strains.We should strengthen virulence genes test and drug resistance monitoring in clinical practice.