Objective To investigate the clinical distribution and antimicrobial resistant characteristics of Pseudomonas aeruginosa(PA)in the northern area of Guangdong.Provide the reference for clinical to prevent infection and reasonable choice of antibiotics and reduce the production of drug resistance strains.Methods The separation and identification of PA were performed by conventional methods during the data of drug 2013 and 2014.The data of sensitivity test of PA were analyzed by WHONET 5.6 and SPSS19.0 softwares.Results The 584 strains PA were mainly distributed in ICU,department of orthopaedics and respiratory medicine.Specimens were mainly from sputum and wound secretion.The detection of PA to 12 antibacterial agents showed different resistance.The antimicrobial with highest resistance was the gentamicin and lower resistance rates to fluoroquinolones,carbapenems,enzyme inhibitors.And a downward trend was shown in drug resistance to CIP,FEP,LEV,SCF.Conclusion PA mainly cause lung and wound infection,especially those old patients that come from ICU,department of orthopaedics and respiratory medicine.Although the drug resistance rates of PA to the commonly used antibiotics are relatively low,The clinicians should reasonably use antibiotics so as to reduce the resistant strains,especially the produce of MDR-PA and PDR-PA.

To predict and identify the dominant B‐cell epitopes of conserved region of Treponema pallidum repeat protein F (TprFN ) and provide the basis for development of polyvalent epitope‐based syphilis vaccine ,the amino acid sequence of TprFN was obtained from GenBank and analyzed with comprehensive meta‐analysis Mobyle ,ABCpred and IEDB online software .The peptides containing predicted epitopes were artificially synthesized . To obtain and measure the titers of antibodies against TprFN ,New Zealand rabbits were immunized with recombinant protein TprFN expressed in E .coli and identified by Western blot (WB) .Sera from TprFN‐immunized rabbits ,syphilis patients ,and normal human and normal rabbits were used to deter‐mine the immunoreactivity and specificity of 7 predicted peptides of TpFN by indirect ELISA .Comprehensive meta‐analysis of online software showed that P1 (43‐62AA) ,P2(57‐71AA) ,P3(81‐88AA) ,P4(89‐103AA) ,P5(125‐138AA) ,P6(231‐251AA) and P7(268‐279AA) might be the B‐cell epitopes .A protein was expressed in a soluble form and identified as TpFN by WB .The ELISA indicated that P1 and P3 were active with TprFN‐immunized rabbit sera and syphilis patient sera but not with negative control sera .These results indicate that P1 and P3 are the potential dominant B‐cell epitopes .

Breaking through the traditional teaching model of medical microbiology,establishing the teaching philosophy of the subject to professional features and the core of combination with form as a breakthrough,promoting the intrest in active learning and the improvement of comprehensive ability of nursing profession students.

Objective To investigate the immune response to and protective effect of a bivalent DNA vaccine expressing interleukin-2(IL-2)and Gpd proteins in New Zealand rabbits.Methods Seventy-two male New Zealand white rabbits were equally and randomly divided into 4 groups to be immunized with recombinant plasmids pcDNA3.1(+)/Gpd-IL-2(pcD/Gpd-IL-2),pcDNA3.1(+)/Gpd(pcD/Gpd),empty plasmid pcDNA3.1(+)(pcD)and phosphate buffered saline(PBS),respectively.Immunization was carried out by intramuscular injection at multiple sites with a 2-week interval for 3 times.On week 10 after the initial immunization,the rabbits were challenged intradermally with T.pallidum(Nichols strain).Enzyme-linked immunosorbent assay(ELISA)was used to quantify the serum level of anti-Gpd antibodies in the rabbits and the level of IL-2 and interferon(IFN-γ)in the supernatant of Gpd protein-stimulated spleen cells from the rabbits at different time pionts.MTT assay was conducted to detect the proliferation response of spleen cells collected from the rabbits on day 0,14,28,140 and 168 after the challenge.Results Compared with pcD and PBS,both the vaccines pcD/Gpd and pcD/Gpd-IL-2 elicited significantly higher levels of anti-Gpd IgG antibodies in rabbits at different time points during the vaccination and infection period,with the titers peaking at 1 ∶ 1024 and 1∶4096,respectively(both P ＜ 0.01).There were also significant differences in the serum levels of anti-Gpd IgG antibodies between the pcD/Gpd-and pcD/Gpd-IL-2-immunized rabbits at different time points(all P ＜0.01).The levels of IL-2 in the supematant of spleen cells from pcD/Gpd-and pcD/Gpd-IL-2-immunized rabbits on week 8 after the immunization were 110 ± 12.6 and 167 ± 15.7 μg/L respectively,and those of IFN-γwere 225 ± 17.6 and 447 ± 22.4 μg/L respectively,significantly higher than those in that from the other two groups of rabbits(all P ＜ 0.01).Furthermore,an apparent proliferation response was observed in spleen cells from pcD/Gpd-and pcD/Gpd-IL-2-immunized rabbits with a higher stimulation index compared with pcD-and PBS-immunized rabbits(all P ＜ 0.01).Dark-field microscopic examination of early-stage infected lesions revealed that pcD/Gpd-IL-2-immunized rabbits had a lower detection rate(17.5％)of Tp from lesions,occurrence of ulcerative lesions(15％)and shorter curing time compared with pcD/Gpd-immunized rabbits.Conclusion The recombinant plasmid pcDNA3.1(+)/Gpd-IL-2 could induce protective humoral and cellular immune response more efficiently in rabbits.

Objective To clone, express Tp0319 gene from Treponemapallidum (T. pallidum), and to assess the immunocompetence of recombinant protein. Methods The immuno-dominant region of Tp0319gene was chosen by computer analysis, amplified from T. pallidum complete genome by PCR, subcloned into the expression vector pQE32 to construct a recombinant plasmid, pQE32/Tp0319, which was then expressed in E. coli M15. The recombinant protein was purified with Ni-NTA affinity chromatography, and identified by using sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) and Western blot. New Zealand rabbits were immunized with the recombinant protein, and the titer of anti-Tp0319 antibodies in sera from immunized rabbits were measured with indirect ELISA. Also, indirect ELISA with the recombinant Tp0319 as coating antigen was performed to detect the anti-Tp0319 antibody in sera from 200 normal human controls and 200 patients with syphilis. Results The prokaryotic expression vector pQE32/Tp0319 was constructed successfully, and the recombinant protein Tp0319 with a molecular weight of about 30 000 was attained. Specific humoral response was elicited by the recombinant protein in New Zealand rabbits and the specific antibody titer was more than 1: 10 240 after immunization for 3 times. Western blot proved that the recombinant protein could specifically react with anti-T. pallidum IgG antibody-positive sera. Indirect ELISA was successfully developed with the recombinant Tp0319, and detected antibodies to T. pallidum in control sera with a sensitivity and specificity of 100% (40/40), respectively. Compared with T. pallidum particle agglutination (TPPA) assay, the sensitivity and specificity of the indirect ELISA were 92.6% and 100%, respectively, in the detection of T. pallidum in sera from patients and controls, and the concordance between the indirect ELISA and TPPA was 96%. Conclusions The prepared recombinant protein shows a satisfactory immunocompetence, which may lay a foundation for its further application in the serodiagnosis of syphilis.

[Objective]To measure the posterior sloope angle of proximal tibial medial meniscus(PSAPTMM) in Chinese adult,provide parameters for improving the design of knee prosthesis more suitable for Chinese and for resecting tibial plateau in total knee arthroplasty.[Method]Sixty-three male knees and 52 female knees of Chinese,without tibial deformity,genu varum and valgum,abnormal change of articular surface,and tibial trauma,with average age of 45.6 years(ranged,20~74 years,were divided into 3 groups according to persons age: group A,18~39 years;group B,40~50 years;group C,more than 60 years.The sagital tomography of medial meniscus of MRI were taken,stored,and measured using JENNY way.The data were analyzed statistically.[Result]By two factors analysis,the sex and age had no significant influence on PSAPTMM,and the angle was not different between the left and right knee.In male adult,the average degree of left PSAPTMM was(2.7?1.8,of the right(2.9?1.7)? with an average degree of both sides of(2.9?1.8.In female adult,the average degree of left PSAPTMM was(3.5?1.7,of the right(2.8?1.6,with the average degree of both sides(3.2?1.7.The average degree of group A were(3.6?1.6,group B(2.7?1.8,group C(2.8?1.7.There showed no significant difference of PSAPTMM between sex,side,and age.[Conclusion]The average degree of PSAPTMM in northern Chinese adult is(3.0?1.7,ranges from(0~6.2,and is larger than that of the westerns.

With the development of basic disciplines such as molecular biology,immunology,cell biology and so on. the pathogen biology research do not stop at the organ and cellular level,but go deep into the protein and gene level. It is a great boost to the deep studies of pathogen biology in diagnosis,treatment,pathogenesis,prevention and epidemiology.

ObjectiveTo evaluate the effect of transureteroscopic Holmium:YAG laser lithotripsy on ureteral calculi.Methods105 cases of ureteral stones were treated by transureteroscopic Holmium:YAG laser lithotripsy. The transverses of stones were 4～15 mm and the vertical lengths were 5～25 mm.ResultsSuccessful lithotripsy was achieved on one session in 98 cases (95%), with the stones completely expelled within 2～6 weeks. Complications occurred in 4 cases and stones moved up in 3 cases.ConclusionThe transureteroscopic Holmium:YAG laser lithotripsy is an effective and safe method for ureteral calculi.

Objective To clone.and express Tp0453 outer membrane protein of Treponema pallidum and develop an indirect ELISA for sero diagnosis of syphilis. Methods The immuno-dominant epitope of Tp0453 was amplified by PCR and subcloned into the expression vector pQE32.The recombinant protein was expressed in E.coli M15 and purified with Ni-NTA affinity chromatography columns. Indirect ELISA was developed to detect the antibody to Tp in human sera.Results 60 control sera was tested by ELISA.The sensitivities was 100%(30/30), and the specificities was 100%. While detecting uninfected and infected T. pallidum human sera, the sensitivities of ELISA was 96.8% compared with the results of the TPPA tests, and the specificities was 100% when the results of ELISA was compared with those of the TPPA test. The concordance of results between the ELISA test and the TPPA test was 98.2%.Conclusion The recombinant Tp0453 outer membrane protein showed excellent immuno-reactive activity, and were suitable for development of ELISA for sero-diagnosis of syphilis.

Objective To investigate the distribution of subtypes of T. pallidum (TP) in Hengyang and Jiangmen regions. Methods Eighty-five specime ns taken from patients with suspected chancre collected in Hengyang and Jiangmen from February 2002 to January 2004 were screened by a PCR targeted TP polA gene , and then the arp gene and tpr gene were amplified from TP positive specimens. The PCR products of tpr gene were digested by restriction endonuclease MSe I. Th e sizes of the arp gene and the restriction fragment length polymorphism (RFLP) of the tpr gene were analyzed for subtyping. Results Of 69 TP-positive specime ns, 57 could be subtyped, and 10 subtypes were found. Among them, 26 (45.6%) wer e subtype 14d, and other subtypes included 10d(1), 12a(3), 12g(2), 13d(6), 14a(5 ), 14b(2), 14f(6), 15d(5) and 16d(1). No significant difference of the distribut ion of TP subtypes between Hengyang and Jiangmen was found. Conclusion Multipl e T.pallidum subtypes have been prevalent in Hengyang and Jiangmen, although the predominant subtype is 14d, there is no significant geographic heterogeneity be tween these two regions.