Objective To investigate the mechanism of cross resistant inducibility of ciprofloxacin and imipenem which led to cross resistance in Pseudomonas aeruginosa in vivo. Methods Cross resistant mutant strains were selected and induced with clinical isolates of P.aeruginosa PA5 susceptible to ciprofloxacin and imipenem by experiment of murine peritonitis. Mutation of DNA gyrase gene, drug uptake and membrance proteins of P.aeruginosa PA5 and its mutants resistant to ciprofloxacin and imipenem were examined. Results Both ciprofloxacin and imipenem could induce resistant strain of P.aeruginosa in experimental murine peritonitis, the cross resistance rates after ciprofloxacin and imipenem challenge were 3.8% and 0.98% respectively. The results of PCR SSCP showed that 3 of six cross resistant of P.aeruginosa strains had gyrA gene mutation. Electrophoresis of outer and inner membrane proteins did not exist any difference between cross resistant strains and their parent strain PA5. Fluorometric assay for ciprofloxacin uptake by bacterial cells indicated that the accumulation of ciprofloxacin in all cross resistant variants decreased to 1/2～1/3 compared with that of PA5. After chanllenge with CCCP, the drug uptake in cross resistance mutants increased to the same level as in PA5. Conclusions The results show that cross resistant strains of P. aeruginosa could be selected and induced in vivo. Active drug efflux is the major factor contributing to the cross resistance of P. aeruginosa to both quinolone and imipenem.

Objective To construct the vector pET15b-Z-VP1 by inserting the Z fragment into amino-terminal of JCV VP1.Methods The VP1 and Z fragment were amplified by PCR from plasmid pET15b and pEZZ18 respectively,and then they were linked by recombinant PCR.The Z-VP1 fragment was inserted into plasmid pET15b by restriction enzyme BamHⅠ and NcoⅠ.Results The VP1 and Z fragment were obtained by PCR and gel purification.The Z-VP1 fragment,which was linked by recombinant PCR from VP1 and Z fragment,was inserted into plasmid pET15b between BamHⅠ and NcoⅠ sites,and confirmed by enzyme digestion and DNA sequencing.The expression of VP1-Z was confirmed by Western blotting.Conclusion The plasmid pET15b-Z-VP1 has been constructed successfully by inserting Z fragment into amino-terminal of VP1.

Objective To express the recombinant protein VP1-Z, and investigate whether VLP-Z has the physiological functions like as wild-type VLP. Methods The expression plasmid pET15b-VP1-Z was introduced into competent E. coil BL21 (DF3)/pLys cells by transformation, and the expression of re-combinant protein VP1-Z was induced by incubation of the cells with IPTG. The protein was prepared as pre-viously described for wild-type VLP. The morphous of VLP-Z were observed by electron microscopy, and the physiological functions of VLP-Z were investigated by hemagglutination test and by immunofluorescence. Re-sults The purified VLP-Z composed of VP1-Z possessed hemagglutination activity and yielded a prominent band of 50×10~3 on SDS-PAGE and staining with Coomassie Brilliant Blue. The VLP-Z exhibited virus-like particles structure like as wild-type VLP with a diameter of 45-50 nm, which was slightly bigger than that of wild-type VLP(42-45 nm). In immunofluorescence test, VP1-Z was detected within the cytoplasm and nu-cleus after HeLa cells were inoculated with VLP-Z. Conclusion The physiological functions of recombinat-ed protein VLP-Z were comparable with wild-type VLP.

Objective To determine pathogenesis and the suitable time of operation for myelomeningocele associated with hydrocephalus in neonate.Methods 6 underwent head CT scanning, 2 lumbosacral CT scanning and 6 lumbosacral X radiography on 6 patients myelomeningocele complicated with hydrocephalus.Ventriculoperitoneal shunt and repair of the myelomeningocele were performed respectively for one patient.from 1 day to 28 day old.Operation stage 1 in 5 patients.Repair of the myelomeningocele concurred with ventriculoperitoneal shunt. Intracranial pressure was measured in shunting procedure.Results 4 patients had normal intracranial pressure,2 patients increased intracranial pressure in the 6 patients.The volume of the hernial sac had markedly diminished and status of hernial sac had greatly improved wall in the patient who wnderwent two-stage procedures after shunt procedure. Lumbosacral wound healing was good . No recurrent myelomeningocele was found, no hydrocephalus was seen using head CT scanning and clinical manifestation has improved in these patients who were followed up 6 month to 3 year.Conclusions Hydrocephalus may deteriorate the degree of lumbosacral myelomeningocele. Effecacy of vntriculoperitoneal shunt and repair of the myelomeningocele was excellent in myelomeningocele complicated with hydrocephalus in neonate.Micro-operative technique might prevent the occurrence of tethered cord.

Adolescent ; Female ; Humans ; Hypersensitivity ; microbiology ; Mitosporic Fungi ; Sinusitis ; microbiology

Objective To investigate the changes of plasma levels of homocysteine (Hcy) and monocyte chemoattract protein 1 (MCP 1) in coronary artery disease (CAD) patients and to analyze the effect of Hcy on the expression of MCP 1. Methods The CAD diagnosis of all the studied subjects was confirmed by coronary angiography. A total of 70 patients with CAD and 50 control subjects were recruited in the study. Plasma Hcy concentration, plasma malondialdehyde (MDA) and plasma MCP 1 were determined by high performance liquid chromatography (HPLC), standard protocols and ELISA, respectively. Results Plasma levels of Hcy, MDA and MCP 1 in patients with CAD were significantly higher than those in the control subjects ( P

AlM:To discuss the feasibility of microcoria optometry in screening for children ametropia. METHODS: Totally 217 school - age children were selected, included 94 first-grade students ( 6 ~ 8 years old) and 123 fourth-grade students ( 9 ~12 years old ) . Refractive diopter was measured with automatic refractor RM-8000 to evaluate the accuracy of micocoria optometry in screening ametropia.RESULTS: After cycloplegia, both the mean sphere diopter and cylinder diopter in grade one students changed significantly (P<0. 05), the mean sphere diopter in grade four students changed significantly (P<0. 05), while the mean cylinder diopter had no statistical difference ( P>0. 05 ) in grade four students. Different refractive type: before and after mydriasis spherical myopia, spherical equivalent difference was 0. 263 ± 0. 618 and 0.216±0.653D, with statistical significance (P<0.01);ln hyperopia group, spherical myopia, spherical equivalent difference was 0. 947±0. 946 and 1. 039±0. 984D, with statistical significance ( P = 0. 000 ). The lenticular difference between the two groups were not statistically different ( P > 0. 05 ). Choosing small pupil computer optometry for ≤- 1. 00D, ≥- 0. 50D child myopia or hyperopia could get more accurate value of diagnostic cutoffs, Youden index was 0. 672 and 0. 580. CONCLUSlON: Microcoria optometry can be as a effective method of screening of children with ametropia, but if for optometry, school-age children must accept mydriasis.

Objective To compare the clinical utility of alpha-fetoprotein (AFP) and des-gammacarboxyprothrombin (DCP) in diagnosing hepatocellular carcinoma (HCC) in patients with a hepatic mass.Methods From January 2015 to May 2015,141 patients were diagnosed to have a liver tumor after imaging examinations in the Hepatobiliary Surgical General Hospital of PLA,Beijing,China.Preoperative AFP and DCP were measured using commercial assay kits.The reference standard was either pathologic or clinical diagnosis of HCC.The performance of AFP and DCP in diagnosing HCC was determined using receiver operating characteristic curve analysis.Results Of 141 patients,98 were diagnosed to have HCC and 43 without.The levels of AFP were significantly higher in patients with HCC than those without [80.0(3.9-1 375.0) μg/L vs.2.1 (1.6-3.2) μg/L,Z =6.98,P ＜ 0.01].Similar results were observed in the levels of DCP [141.5 (24.0-978.0) AU/L vs.19.0 (14.0-25.5) AU/L,Z =5.18,P ＜ 0.01].Receiver operating curves (ROC) indicated the cut-off value with the best sensitivity and specificity was 3.6 μg/L for AFP and 35 AU/L for DCP.The difference in the area under ROC between AFP and DCP was not statistically significant (0.87 vs.0.78,Z =1.72,P =0.085).The sensitivity and specificity for detection of HCC in patients with a hepatic mass were 56.1％ and 95.4％ for AFP ＞ or =20 μg/L,69.4％ and 83.7％ for DCP ＞ or =40 AU/L,respectively.The level of AFP was associated with DCP in patients with HCC (x2 =9.12,P ＜ 0.01,r =0.292) and parallel testing of AFP and DCP gave an optimal sensitivity of 79.6％ with a specificity of 81.4％ in diagnosing HCC.Conclusions DCP is a useful biomarker and it gave an equal performance as AFP in diagnosing HCC in patients with a liver mass in this study.Parallel testing of AFP and DCP effectively increased the diagnostic sensitivity.Although the biomarkers only marginally improved the diagnostic results,it could be useful in diagnosing HCC in individuals who had atypical imaging results.

Objective To investigate the relationship between single nucleotide polymorphisms (SNPs) of PRKCG gene (rs2547362,rs3745406) and osteosarcoma susceptibility in the osteosarcoma patients and the normal population.Methods Sixtyone patients with osteosarcoma who had been admitted in our hospital from January 2011 to December 2012 and 63 healthy adults were enrolled in this study.A 2-ml peripheral blood sample was taken from each participant.The RT-qPCR method was used to detect the genotype and allele frequency distribution of PRKCG gene at rs2547362 and rs3745406 in osteosarcoma patients and normal population.Osteosarcoma patients were divided into several groups according to the clinical parameters such as age,gender,histology,tumor location,Enneking classification,tumor metastasis and therapy,and then we analyzed the relations between the genetic polymorphism and clinical parameters.Results 1) The genotype of PRKCG gene at rs3745406 included CC,CT and TT.The differences of genotypes (CC,CT,TF) and alleles (C,T) frequency distribution at rs3745406 were not statistically significant between osteosarcoma patients and the normal population (P=0.490,P=0.554).2) The genotype of PRKCG gene at rs2547362 included CC,CT and TT.The differences of genotypes (CC,CT,TT) and the alleles(C,T) frequency distribution at rs2547362 were statistically significant between the osteosarcoma patients and the normal population (P=0.006,P=0.007).3) The differences of genotypes (CC,CT,TT) and alleles (C,T) frequency distribution at rs3745406 were statistically significant between patients with metastasis and patients without metastasis (P=0.000,P=0.000).The CT and TT genotypes and the T allele carrier frequency at rs3745406 were higher in patients with metastasis than in patients without metastasis.SNPs at rs2547362 were not associated with clinical parameters.Conclusion The genetic polymorphism of PRKCG gene at rs2547362 is associated with osteosarcoma susceptibility.The TT genotype and T allele at rs3745406 are associated with metastasis of osteosarcoma,which may be a risk factor for metastasis in the osteosarcoma patients.