Objective To investigate antimicrobial resistance characteristics and mechanisms of imipenem-resistant Pseudomonas aeruginosa (P .aeruginosa)isolated from clinical specimens.Methods Bacterial strains were identi-fied by BD Phoenix 100 automatic microbial analysis system,antimicrobial susceptibility testing was performed by Kirby-Bauer method;Carbapenemase genes (IMP ,VIM ,OXA,GES )and outer membrane protein gene oprD2 were detected by polymerase chain reaction.Results Resistant rates of imipenem-resistant P .aeruginosa strains to ami-kacin was the lowest (8.33%);resistant rates to gentamicin and tobramycin were<20%;resistant rates to the third and fourth generation cephalosporins,aztreonam,meropenem,compound sulfamethoxazole and minocycline were all>60%,and all strains were resistant to ampicillin/sulbactam.The positive rate of OXA-17 gene was 2.78%(n=1 ),deletion rate of oprD2 was 38.89%,the other drug-resistant genes were not detected.Conclusion Except aminoglycosides,resistance of imipenem-resistant P .aeruginosa to other antimicrobial agents is serious;resistance of P .aeruginosa to imipenem may be due to lacking of OprD2 and production of carbapenemases.

Objective To study the expression and clinical significance of TNF related apoptosis-inducing ligand(TRAIL) and Caspase-3 in the bone marrow mononuclear cell(BMMNC) of aplastic anemia(AA) ,and to approach their effect on the apoptosis of haemopoietic stem cell in AA patients.Methods The TRAIL and Caspase-3 expression in 15 cases with newly diagnosed AA patients and remission patients,and 15 cases with normal bone marrow patients were detected by flow cytometry and microplate reader. Results The TRAIL expression of newly diagnosed AA patient, emission AA patients and control group was ( 5.94 ± 2. 57 ) ％, ( 2. 87 ± 1. 72 ) ％ and ( 3.01 ±2. 06)％ respectively,the Caspase-3 expression of newly diagnosed AA patient, emission AA patients and control group was ( 1. 3840 ± 0. 0236), (0. 8018 ± 0. 0126) and ( 0. 7441 ± 0. 0112 ) respectively. The TRAIL and Caspase-3 expression of newly diagnosed AA patient were significantly higher than the remission AA patients and control group,the difference was significant( all P ＜0. 05) ;The TRAIL and Caspase-3 expression of control group were higher than the remission AA patients, but the difference was not significant ( all P ＞ 0. 05 ). Conclusion The TRAIL and Caspase-3 had high expression in the BMMNC of AA patients, and the apoptosis induced by TRAIL and Caspase-3 should play an important role in the pathogenesis of AA.

20?mm?Hg, nonpulmonary hypertension group (control group n=9), with preoperative mean pulmonary artery pressure

Ticks transmit various diseases to livestock and human beings. The researches on ticks are of great importance in medical and veterinary sciences. To express a glutathione peroxidase gene (HlGPx) from Hemaphysalis longicornis in Escherichia coli (E. coli) so as to provide basis for further studies, the gene was amplified from cDNAs of H. longicornis adult ticks by PCR prompted by information from EST library. The TGA codon encoding selenocysteine in the gene was mutated into the universal genetic code for cysteine,TGC, by site-directed mutagenesis. E. coli was transformed with the recombinant expression vector pGEX-4T-1/HlGPx'(the mutated HlGPx) and induced to express recombinant HlGPx', which was then analyzed by sodium dodecylsulphate-polyacrylamide gel electrophoresis and Western blotting. The result showed that the complete coding sequence of HlGPx was obtained successfully, the deduced polypeptide was 223 aa, with a calculated molecular mass of 25.0 kDa; the recombinant fusion protein was approximately 51 kDa, correspondent to the calculated. The antibody against glutathione S-transferase (GST) recognized the recombinant protein fused with GST in a Western blotting assay, which confirmed the result above-mentioned. In conclusion, the mutated HlGPx was expressed in E. coli successfully and it could be used for further preparation of immune serum and functional analysis.

Objective To evaluate whether the comparability of 3 automatic blood cell analyzers meet the clinical requirements by conducting the comparative study on the detection results of these instruments.Methods With the Sysmex 2100 automatic blood cell analyzer as the reference instrument,Sysmex 1000i and Abbott 1800 as the experimental instrument,the original quality control provided by the instrument factory and the patient′s fresh anticoagulant blood samples in the laboratory were adopted to monitor for continuous 40 d by these three instruments and the detection results of WBC,RBC,HGB,HCT and PLT were analyzed.Results The detection results of these 3 instruments were statistically tested by the F test,the differences showed no statistical significance (P >0.05)and the bias was in 1/2 of the maximum permissible error range in America department clinical test revised regulations (CLIA′88).Conclusion The detection results by these 3 instruments are comparable and can meet the clinical requirements.

Background Triamcinolone acetonide (TA) is often used in the treatment of uveitis and fundus disease,but whether it has toxic effect to normal tissue around eyes is unclear.Objective The aim of this study was to explore the adverse and toxic effects of TA on the periorbitally normal tissue following the periocular injection.Methods Twenty-seven New Zealand albino rabbits were randomly divided into experimental group,normal saline solution group and blank control group.TA was periocularly injected twice (20 mg for each) at 1-month interval in the rabbits of the experimental group,and 0.5 ml normal saline solution was used in the same way in the rabbits of the normal saline solution group.Not any drug was used in the blank group.The rabbits were sacrificed 1 month and 2 months,3 months after the secondary injection.For the preparation of the specimens of extraocular muscles,peribulbus adipose tissue,lacrimal gland and optic nerve.Hematoxylin and eosin staining of the specimens was performed to examine the the pathological change under the optical microscope,and the ultrastructural of peribulbus tissues was observed under the transmission electron microscope.Results No statistically significant change was found in IOP among various time points and groups (Fgroup =0.952,P =0.881 ; Ftime =7.297,P =0.411).The hyphological structures and ultrastructure of extraocular muscles,peribulbus adipose tissue,lacrimal gland and optic nerve were normal in the rabbits of the normal saline solution group and the blank control group.However,the optical microscopy showed varying degrees of atrophy or dissolving in the extraocular muscle fibers,the increase of glandular epithelial cells and enlargement of nuclei in the lacrimal gland as well as visible disorder arrangement of nerve fibers,vacuoles degeneration and the decrease of glial cells in optial nerve tissue in the rabbits of the experimental group from 1 month though 3 months after the secondary injection.Under the transmission electron microscope,the muscle stripes,bright band and dark band were incompletely disappeared,and expansionary sarcoplasmic reticulum and oval nucleus at the inferior to sarolemma were seen in the extraocular muscles.In addition,swelling lacrimal gland epithelial cells,thickening nuclear membrane and more organelles were exhibited in the lacrimal gland.In the optic never tissue,uneven myelin,space between axons and myelin,lamellar separation and degeneration also were revealed in the rabbits of the experimental group after injection of TA.Peribulbus adipose tissue was near normal both by optical microscope and transmission electron microscope in the experimental group.Conclusions The periocular injection of TA dose not elevated the IOP.However,it will result in the histogical and ultrastructural damages of extraocular muscles,lacrimal gland and optic nerve.

ObjectiveTo test the expression of epidermal growth factor-like domain 7 (EGFL7),microvessel density (MVD) and foeal adhesion kinase pY397 (FAKpY397) in human glioma tissues,and to evaluate their relationship.MethodsThe expression of EGFL7 and FAKpY397 in 56 cases of human glioma and 8 cases of normal brain tissues were detected by immunohistochemistry test,and MVD was detected by CD34 staining.ResultsThere was a significant difference of the positive rates of EGFL7 between normal brain tissue (0) and gliomas (75％),χ2 =17.45,P ＜0.01.With the increased pathological grade,the expression level of EGFL7 increased (χ2 =26.24,P ＜ 0.01 ).There was a significant difference of the positive rates of FAKpY397 between normal brain tissue ( 12.5％ ) and gliomas (73.2％),χ2 =6.23,P ＜ 0.05.With the increased pathological grade,the expression level of FAKpY397 increased (χ2 =6.71,P ＜ 0.01 ). MVD on normal brain was( 15 ± 4 )/HP,on Ⅰ - Ⅱ grade and Ⅲ -Ⅳ grade gliomas was ( 27 ± 3 )/HPand ( 60 ± 4 )/HP respectively,there was a significant difference on MVD between normal brain tissue and gliomas (P ＜ 0.01 ).Higher level of MVD was found in gliomas with higher grade ( P ＜ 0.01 ).There was a positive correlation between EGFL7 and FAKpY397 expressions in gliomas (r =0.314,P ＜0.01 ).There was a significant difference on MVD between positive and negative expression of EGFL7 ( t =26.55,P ＜ 0.01 ). MVD was (56 ± 4 )/HP and (25 ± 3 )/HP respectively.ConclusionThe expression of EGFL7 of human gliomas has a favorable positive correlation with the degree of malignancy,MVD and FAKpY397.It is indicated that EGFL7not only palys an important regulative role in glioma neovascularization,but also it may participate directly in glioma occurrence and invasion.

Objective To provide the scientific basis for realizing the inter-accreditation of laboratory electrolyte detection results by comparing the electrolyte detection results in 5 grade 3A hospitals of Zhuhai city .Methods Each 10 serum samples with low , middle and high concentrations of electrolyte were collected for simultaneously detecting the electrolyte kalium (K) ,natrium (Na) and chlorinum (Cl) .The detection results were performed the statistical analysis and comparison .The mutual bias within 1/2 of al-lowable error of CLIA′88 indicated that the detection results were mutually accredited ,if the mutual bias exceeding 1/2 of allowable error ,the detection results could not be mutually accredited .Results The difference of electrolyte detection results in 5 hospitals accorded with the stipulation requirement of CLIA′88 .Conclusion The electrolyte detection results of 5 hospitals could be mutually accredited .

BACKGROUND: Studies have shown that hemiarthroplasty is suitable for single femoral head diseases with normal acetabulum. But indications for choosing bone cement or biological fixation types, short- and long-term efficacy as well as the complications have not yet reached agreement.OBJECTIVE: To discuss the efficacy of two kinds of femoral prosthesis for femoral neck fractures in hemiarthroplasty,and the application of three-dimensional CT reconstruction in displaced fractures.METHODS: Clinical data of 73 cases of femoral head fractures undergoing hemiarthroplasty were analyzed retrospectively, and divided into bone cement and biological groups. All patients were followed up for 1.5 years, and the operation indexes, hospitalization time and hip joint function were compared; the consistent degree with intraoperative findings was compared through hip three-dimensional CT and X-ray examination simultaneously.RESULTS AND CONCLUSION: (1) There was no significant difference in the effective rate between groups (P > 0.05).(2) The operation time in the biological group was significantly shorter than that in the bone cement group (P < 0.05); the intraoperative blood loss and hospitalization time in the bone cement group were significantly less than those in the biological group (P < 0.05). (3) The Harris scores in the bone cement group were significantly higher than those in the biological group at 1 month after replacement (P < 0.05); the Harris scores in the biological group were significantly higher than those in the bone cement group at 18 months after replacement (P < 0.05); while there were no significant differences between two groups at 3, 6 and 12 months after replacement. (4) The hip three-dimensional reconstruction and X-ray examination showed no error compared with the intraoperative findings, reaching 100%. (5) Our findings suggest that age, disease history and bone quality should be considered when choosing an appropriate type of hemiarthroplasty for the patients with femoral neck fractures, and bone cement prosthesis is suitable for the elder patients with severe osteoporosis; while those with good bone quality should receive biological prosthesis. Besides, the diagnosis accuracy of three-dimensional reconstruction is superior to X-ray examination, which is conductive to fully understand displaced fractures.

Objective:To provide a potential platform for transferring specific antigen against fish bacterial diseases based on attenuated Lm (EGDe-ΔactA/ΔinlB).Methods: Attenuated Lm (EGDe-ΔactA/ΔinlB) was used to express outer membrane protein K (Ompk),a conserved and effective vaccine candidate in vibrio.The identification of recombinant strains and detection of antigen genes were operated with PCR and RT-PCR,respectively.Results: The results of PCR showed that Lm-Ompk (L-O),Lm-Lmo0576-Ompk (L-L-O) and Lm-P-Ompk (L-P-O) were constructed successfully.The identity of foreign gene was 100％ compared with sequence of NCBI.The analysis of transcription showed that the expressions of Ompk in L-O,L-L-O and L-P-O were significant (P<0.001).Moreover,the expression of Ompk in the condition of antibiotic was higher than that in the BHI without antibiotic (P<0.05).Conclusion: Lm-Ompk (L-O),Lm-Lmo0576-Ompk (L-L-O) and Lm-P-Ompk (L-P-O) were constructed successfully.