Objective To prepare recombinant plasminogen activator(Pla) protein in E.coli BL21 cells that can be used in studying interactions between Yersinia pestis proteins and immunologic diagnosis of plague.Methods The pla gene was amplified by PCR and cloned into the pET28a expression vector.E.coli BL21 competent cells were transformed with the recombinant vectors, and isopropyl-β-D-thiogalactopyranoside ( IPTG) was added to induce expression of Pla protein. The expressed protein was detected by SDS-PAGE electrophoresis.The inclusion bodies of Pla protein were denatured in 8 mol/L urea, and then refolded using gradient urea solutions.The purified protein was identified by SDS-PAGE electrophoresis and Western blot.Results and Conclusion The constructed expression vector was demonstrated to be correct through agarose gel electrophoresis and sequencing.The recombinant Pla protein was accumulated as an inclusion body in E.coli, and the overexpression product was mainly a target protein, the yield of which was very high.SDS-PAGE purity of the bioactive Pla protein was obtained by denaturing and refolding the inclusion bodies.This study provides a simple and quick method for highly efficient preparation of biologically active Pla protein.

The gene of mutated TNF-?D4 gene was amplified by overlap PCR and cloned into the prokaryotic expressive vector pBV220.TNF-?D4 contains two changes:substitutions of Pro8Arg,Ser9Lys,Asp10Arg,Ile157Phe,Leu29Ser,Arg31Val and a deletion of the N terminal four amino acids.The recombinant vector pBV220-TNF-?D4 was transformated into E.coli strain DH5?,and the high expression strain was obtained by screening monoclones.The level of expression was about 45% of total cell protein.After purification,the purity of fusion protein was above 90% by HPLC and relative ability was 8 ?107.TNF-?D4 was modificated by mPEG-ButyrALD。After purification,the purity of mPEG-TNF-?D4 was above 85% and relative ability was 8.6?107.The in vivo systemic toxicity of mPEG-TNF-?D4,which is indicated by LD50,is lower than that of rhTNF-?.These results strongly supported for the further study and exploitation of TNF-antitumor drug.

Objective To establish a simple and quick loop-mediated isothermal amplification (LAMP)method for detection of Yersinia pestis.Methods LAMP Primers were designed based on the specific sequence 3a in Y.pestis chromosome.LAMP reaction results were detected using turbidity meter or visual method.The specificity of the constructed method was evaluated by detecting Y.pestis and its closely-related bacteria.The different dilution DNA template was detected with LAMP and PCR to evaluate the sensitivity of the method.Results Thirty strains of bacteria closely related to Y.pestis were detected by the constructed LAMP,and all the results were negative,indicating that the method had a very high specificity.The detection sensitivity of this LAMP assay was 20 pg of DNA per reaction,which was ten-fold that of the regular PCR.The detection reaction was completed in 25 min.Conclusion This LAMP method is quick,sensitive, specific and simple,which is expecked to become an effective method for rapid detection of Y.pestis on the scene.

Objective To explore the clinical application of first class nursing evaluation list in patients with osteoporotic vertebral compression fractures after surgery.Methods Totally 100 patients with osteoporotic vertebral compression fractures hospitalized in the Department of Spinal Surgery of our hospital were selected from February 2014 to February 2016 by convenience sampling. 50 patients selected from February 2014 to January 2015 were divided into the control group, while 50 patients selected from February 2015 to February 2016 were divided into the observation group. Patients in the control group received routine nursing evaluation sheet for inspecting and supervising the implementation of postoperative nursing measure, while patients in the observation group also received the postoperative first class nursing evaluation list about osteoporotic vertebral compression fractures for inspecting and supervising the implementation of postoperative nursing measures. The ability of daily life, nursing satisfaction and quality of care of patients between the two groups were compared.Results The Barthel index score of patients in the observation group one week after surgery was (84.39±12.75), health education in nursing satisfaction (4.31±0.65), service attitude (4.36±0.31), nursing process (4.38±0.33), nursing effect (4.12±0.41), the nursing measure omission rate 4%, and the pass rate of quality of care 98%; while in the control group, the Barthel index score was (50.15±9.89) (one week after surgery), health education (3.46±0.58), service attitude (3.7±0.50), nursing process (3.75±0.52), nursing effect (3.34±0.47), nursing measure omission rate(30%), the pass rate of quality of care 80% (P<0.05). Conclusions Application of first class nursing evaluation list can greatly improve the recovery situation of patients with osteoporotic vertebral compression fractures, and it also can improve the ability of daily living and nursing quality, so it is worth clinical promotion.

Objective To establish competent characteristic system and a level standard model for orthopedic specialized nurses.Methods From October 2016 to March 2017, a total of 40 medical and nursing specialists from 10 provinces and cities were selected by convenience sampling methods as specialist consultants. Two rounds of Delphi specialist consultation were conducted on 7 primary indexes and 58 secondary indexes of the competent characteristic system for orthopedic specialized nurses which was primarily confirmed by literature search. The index names were modified, merged or deleted according to experts' opinion and statistical analysis results. At last, 12 experts in Beijing area were selected, and the typical behavior event interview method was adopted to determine the level standards of the secondary indexes.Results The experts' authority coefficient was 0.91. The effective recovery rates of questionnaires of the two rounds expert consultation were 95.00％ and 92.11％ respectively. The primary indexes consists of 7 indicators: clinical decision (0.1500), nursing practice (0.1543), management (0.1356), communication(0.1422), legal ethics(0.1289), professional development (0.1364) and education(0.1322). Twenty-eight items of secondary index were confirmed, and Likert Scale was applied to determine the level standards of secondary index and describe typical behavior. Level 1 to 5 of Likert Scale was corresponding to the levels to orthopedic specialist nurse's career planning respectively, including entry-level, qulified, competent, proficient, and expert level.Conclusions The evaluation indicators recommended in specialist consultation meet the specialized nurse competent characteristics, and quantitative evaluation of specialized nurses can be achieved by the level standard system. The evaluation and level standard system can analyze the strengths and weaknesses of the subjects, to achieve evaluation, education, management and career planning for orthopedic specialized nurse.

Objective To discuss the clinical effect of medial canthal skin defects reparation with nasofacial sulcus reversed fascio-cutaneous flaps.Methods Medial canthal skin defects in 10 patients were repaired with nasofacial sulcus reversed fascio-cutaneous flaps in the hospital.These patients included 3 cases of pigmented nevus,6 cases of skin tumors,1 case of scar.The size of defects varied from 0.8 cm× 1.0 cm to 1.8 cm×2.2 cm.Results In these 10 cases,the maximum area of fascio-cutaneous flaps was 2.0 cm×3.0 cm,and the minimum area was 1.2 cm× 1.5 cm.All the flaps survived with primary healing postoperatively.Incision line scar was inconspicuous and the color and texture of flaps were similar to those of surrounding tissues.Conclusions To maintain and restore the integrity of medial canthal by using nasofacial sulcus reversed fascio-cutaneous flaps would lead to a good aesthetic effects.

OBJECTIVETo investigate the significance of Rac subfamily members in the gastrointestinal carcinogenesis and progression.

METHODSThe mRNA expression of Rac1, Rac2 and Rac3 in 12 kinds of gastrointestinal cancer cell lines was examined by semi-quantitative RT-PCR. The activities of Rac1 protein in 5 kinds of gastric cancer cell lines were tested by pull-down assay.

RESULTSCompared with the normal gastric mucosa and intestinal epithelial cell line, the mRNA expression of Rac1 and Rac3 was up-regulated in most of gastrointestinal cancer cell lines. The activities of Rac1 protein increased markedly in gastric cancer cell lines.

CONCLUSIONThe increased mRNA expression of Rac1 and Rac3 in gastrointestinal cancer cell lines and the abnormal activation of Rac1 protein in gastric cancer cell lines might be correlated with the carcinogenesis of gastrointestinal cancer.

Cell Line, Tumor ; Gastrointestinal Neoplasms ; metabolism ; Humans ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction ; rac GTP-Binding Proteins ; genetics ; rac1 GTP-Binding Protein ; analysis ; genetics

Objective:To explore the clinical application of orbital septum fat reset for tear though deformity and palpbromalar groove in the lower blepharoplasty.Methods:From March 2015 to March 2018, 72 cases of lower eye baggy deformity with tear though deformity and palabromalar groove were treated with the procedures of orbital septum fat reset, relaxing the tears ligament and orbicularis retaining ligament; the orbital fat that was released from orbital septum was repositioned at the inferior orbital and fixed on periosteum.Results:The incisions of 72 cases were healed successfully, the shape of the eyes was naturally symmetrical, the palpebral and buccal parts were excessive, and the contour lines formed a monoconvexcurve, with no obvious scar. All the cases were available for 6 to 24 months follow-up. 66 cases (91.67%) were satisfied with the cosmetic results.Conclusions:Orbital septum fat reposition is an effective method for correction of tear though and eyelid sulcus deformity.

Objective To evaluate the effect of round going process of nursing group leader in Orthopedic Department.Methods The round going process of nursing group leader in orthopedic department had been implemented since January 2016 in the 309th Hospital of PLA. It was compared, before ( March 2015) and after ( March 2016) the implementation, of the patients′ awareness of health education, qualified rate of basic nursing quality, accuracy of nursing assessment and high risk assessment, patients′satisfaction, accuracy rate of nursing marks and implementation of checking and signature. Results Before and after the implementation, of patients′health education awareness, rates of awareness of specialty drugs were 92.50% and 98.33% and rates of perioperative health education were 94. 17% and 99. 17%; of qualified rates of basic nursing, rates of cleanliness were 76.30% and 91.11% and rates of comfort were 81.11% and 92.22%; of nursing assessment accuracy, rates of admission assessment were 93. 33% and 98. 52% and rates of high risk assessment were 91.67% and 98.33%; of accuracy of nursing marks, rates of nursing grade were 85. 88% and 96. 68%, rates of tube marking were 80.72% and 92.64%, and rates of high risk marking were 95.74% and 98.89%; of implementation of checking and signature, rates of patrol card were 83. 17% and 95. 26%, rates of nursing measure table were 84.95% and 97.67%, and rates of turning over card were 89.24% and 95.63% ( P<0.05) . Conclusions Process quality control, health education level and basic nursing quality can all be improved by round going process of nursing group leader in orthopedic department.

Objective To comprehend status quo of nursing assessment of duty nurses in orthopedic department and to develop corresponding strategies, so as to provide references for clinical teaching of specialty nursing.Methods In March 2016, 48 duty nurses in the Orthopedic Center in The 309th Hospital of Chinese PLA , were selected by convenience sampling method and examined about their nursing assessment, using the self-designed nursing assessment process and criteria. Comprehensive assessment was done by rank-sum ratio in CHISS 2011, and corresponding strategies were make accordingly.Results Analysis result of the rank-sum ratio showed that, 32 items could be divided into three levels according to times of mark deduction, with 7 items of the first level, 18 of the second, and 7 of the third. The first level included specialty physical examination, inquisition of the illness history, inquisition of current symptoms, assessment method of health education effect, induction of positive data, making nursing goals and putting forward nursing diagnosis. Conclusions In response the above mentioned problems, Nursing handbook of specialty physical examination in orthopedic department, Nursing inquisition guidelines of duty nurses in orthopedic department, and Standard operating procedure on nursing health education in orthopedic department were made. Layered training examinations were conducted accordingly, so as to improve nursing assessment quality of duty nurses.