Objective To cultivate and identify the transgenic affalfa containing Echinococcus granulosus Eg95 gene. Methods The alfalfa plants were transformed by co-cultivating alfalfa cotyledons via recombinant Agrobacterium tumefaciens LBA4404 harboring pBI-Eg95. The transgenic alfalfa explants were selected by kanamyein after calli formation, shoots and roots regeneration in the selective medium, the seedlings of transgenic plants were obtained which were finally transplanted into pots containing nutrient soil. After 2-3 months growth, the complete transgenic alfalfa plants containing Echinococcus granulosus Eg95 gene were obtained. To identify the transgenic alfalfa plants, the total DNA, RNA and leaf protein were extracted from fresh leaf tissue of the transgenic alfalfa plants and confirmed by PCR, RT-PCR, SDS-PAGE and Western blot assay. Results A specific band around 471 bp was amplified by PCR with total DNA, and the same band was obtained by RT-PCR with total RNA, which confirmed that the Eg95 gene was stably integrated into the transformed alfalfa genome. SDS-PAGE analysis showed that the relative molecular mass(Mr) of the expressed protein was about 16.5×103, consistent with the Eg95 protein, and the level of Eg95 expression was up to 0.06% of total soluble leaf protein by Bio-Rad Quantity one assay. Western blot verified the expressed protein was reactive with the sera of mice infected with Echinococcus granulosus. Conclusion The transgenic alfalfa plants containing Echinococcus granulosus Eg95 gene are successfully cultivated.

Objective To understand the differences of virulence genes and molecular typing in Campylobacterjejuni isolates from poultry products and diarrhea patients in Shenzhen.Methods According to specific primers,four virulence genes (cdtB,cadF,flaA,virB1 1 )of C.jejuni were detected by polymerase chain reaction (PCR).Molecular typing for C.jejuni strains was performed by Pulsed-field gel electrophoresis (PFGE).Results There were no differences of gene distribution (cdtB+,cadF+,virB1 1-)between isolates from poultry products and diarrhea patients.Two virulence genes of cdtB and cadF were found in all of ten C.jejuni strains lacking virB1 1 .The carriage rate of flaA in food-borne isolates (3/5 )was higher than those in patient isolates (2/5).In the PFGE map,the clustering analysis of C.jejuni strains showed that a total of 5 to 9 DNA bands were observed in ten strains through the digestion of Sam I.There was high homology (above 85%) between food-borne isolates and patients isolates,but the distribution of flaA in these highly homologous strains was differ-ent.Conclusion So far,C.jejuni strains with cdtB,cadF and flaA were present in Shenzhen,and showed high diversity and homology.This implies that the occurrence of diarrhea in patients with C.jejuni was associated with the contaminated poul-try products by this pathogen.Their findings can provide basic data and evidences about diarrheal disease caused by food-borne C.jejuni for the local region.

BACKGROUND:Taurine is an important non-enzymatic system antioxidant in the lens.The mechanism of its anti-oxidative effect is mainly to protect lens from oxidative injury by anti-lipid peroxidation.OBJECTIVE:This study was to observe the effect of exogenous taurine on lens epithefial cell apoptosis-induced by H2O2 in vitro.DESIGN:A randomized controlled animal experiment.SETTING:Department of Ophthalmology,Affiliated Hospital of Qingdao University Medical College.MATERIALS:Seventy-five adult New Zealand standard tabbits,of either gender,weighing 1.5-2.5 kg,were provided by Qingdao Laboratory Animal Center.Reagent kit for in situ detecting cell apoptosis(Sigma Company,USA),taurine and H2O2(Shanghai Guangda Chemical Reagent Factory,China)were included in this study.METHODS:This study was performed in the Department of Ophthalmology and Central Laboratory.Affiliated Hospital of Qingdao University Medical College from Mav 2005 to June 2007.Rabbit lenses were harvested and randomly divided into 3 groups:control group,in which,clear lenses were incubated in non-serum and non-phenolsulfonphthalein MEM medium which was renewed every 24 hours,H2O2 group,in which,clear lenses were incubated in non-serum and non-phenolsulfonphthalein MEM medium containing 1 mmol/L H2O2 with addition of 62 μL H202(30 g/L)every 6 hours,and H2O2+taurine group,in which,clear lenses were incubated in non-serum and non-phenolsulfonphthalein MEM medium containing 1 mmol/L H2O2 and 10 g/L taurine.which was renewed every 6 hours.The protocol was conducted in accordance with ethical guidelines for the use and care of animals.MAIN OUTCOME MEASURES:Observation of lens opacity 6.12,24.48 and 72 hours after culture;Lens epithelial cell apoptosis determined by DNA in situ end labeling and DNA fragment analysis.RESULTS:Lens opacity:The lens opacity in the H2O2 group was aggregrated gradually along with the time of oxidative injury.The lens opacity in the H2O2 group was severer than that in the H2O2+taurine group.Lens epithelial cell apoptosis:There were no apoptotic cells in the control group within 72 hours.The number of apoptotic cells in the H2O2 group was increased gradually with the prolonged time of oxidative injury.Till the 72nd hour,the cells were all tamed into apoptotic cells.A few aopototic celIs were found in the H2O2+taurine group since hour 24,and then were more and more,and the number of apoptotic cells accounted for about 30%at hour 72.The apoptotic rate in the H202+taurine group was significantly lower than that in the H2O group at each time point(q=8.6845,P＜0.01).There was no significant difference in apoptotic rate between the H202+taurine group and the control group (P＞0.05). Findings of DNA fragmentation assay:The DNA"ladder"was found in the H2O2 group at hours 24,36,48 and 72,while no DNA"ladder"but only normal electrophoresis straps were present in the other two groups 24 hours after culture.CONCLUSION:Taurine can inhibit the oxidative injury-induced apoptosis of rabbit lens epithelial cells,and alleviate lens opacity.

Aspergillosis, Allergic Bronchopulmonary ; diagnosis ; drug therapy ; Child ; Humans ; Male

Abstract: Objective　To analyze the clinical characteristics of Chlamydia psittaci pneumonia and improve the diagnosis and treatment skills of clinicians on this disease. Methods The clinical data of thirty-nine Chlamydia psittaci pneumonia cases detected by metagenomic next-generation sequencing (mNGS) from September 2020 to January 2022 at the Affiliated Hospital of Southwest Medical University were retrospectively analyzed. Results There was a history of poultry exposure in 89.7%(35 cases) of the patients. The most common clinical manifestations were high fever (92.3%, 36), cough (76.9%,30), muscle soreness (48.7%,19), headache (38.5%,15), etc. Laboratory examinations showed 76.9% of patients had a normal leukocyte count, and 76.9% had decreased lymphocyte count, often accompanied by elevated C-reactive protein (100%), procalcitonin (97.4%), interleukin-6 (95.8%), interleukin-10 (95.8%), alanine aminotransferase (74.4%), and aspartate aminotransferase (84.6%). Univariate analysis indicated that there were statistically significant differences in the levels of aspartate transaminase, blood urea nitrogen, C-reactive protein, and procalcitonin between severe pneumonia patients and non-severe pneumonia patients（P<0.05）. Multivariate logistic regression analysis showed that an elevated blood urea nitrogen (OR=4.899) had guiding significance for predicting the occurrence of severe pneumonia. Bronchoscopy examination showed no abnormalities in 53.6% of the patients. The imaging manifestations of pulmonary lesions were mainly lobar pneumonia (61.5%) and air bronchograms (94.9%). Therapeutically, it was sensitive to tetracyclines, macrocyclic lactones, and fluoroquinolones. A total of 84.6%(33 cases) of the patients were cured and discharged from the hospital at the end of the treatment. Conclusion Chlamydia psittaci pneumonia is a zoonotic disease that can be detected by mNGS. An elevated blood urea nitrogen level has guiding significance for predicting the occurrence of severe pneumonia. Empirically-selected regimens based on doxycycline are effective for the treatment of Chlamydia psittaci pneumonia.

BACKGROUND:Cerebral aneurysm is a kind of mortal hemangioma, and its treatments such as endovascular embolization and clipping both cause high postoperative recurrence rate and mortality. So the stent implantation for cerebral aneurysm is coming into being. OBJECTTVE:To evaluate the hemodynamic parameters after stent implantation into cerebral aneurysm and to provide a novel feasible strategy for clinical treatment. METHODS:A retrospective analysis was preformed based on the CT image data of 11 patients with cerebral aneurysm from the Affiliated Hospital of Xinjiang Medical University. Firstly, the flexible and solid model of cerebral aneurysm was established by the MIMICS and reverse engineering. Secondly, the matching stent model was implanted into the cerebral aneurysm, and then the blood flow structure of cerebral aneurysm was analyzed by the fluid dynamics theory and the Fluent with the method of two-way flow solid coupling. Final y, comparative analysis of the kinetic parameters of cerebral aneurysm before and after implantation, including wal pressure, blood velocity, path line of the blood flow, wal shear stress, wal deformation was conducted, and blood flow characteristics after stent implantation were analyzed under different entrance velocity. RESULTS AND CONCLUSION:After implantation, the wal surface pressure was reduced about 61.1%;the blood flow velocity around the stent and the inside of the cerebral aneurysm was decelerated obviously;under setting 2 000 lines of blood flow, the number of path line of blood flow into the cerebral aneurysm reduced about 75.0%, the maximum wal shear stress decreased about 79.3%, and the maximum wal deformation reduced to a lower level. The entrance velocity was respectively v1=0.1 m/s, v2=0.2 m/s, v3=0.3 m/s and the wal pressure was in a gradient ascent;the wal shear stress increased with the velocity, meanwhile,τzou (left neck of aneurysm)<τzhong (aneurysm )<τyou (right neck of aneurysm). The path lines of blood flow mainly concentrated in the top of the aneurysm, and the blood velocity markedly affected the surface deformation. These results indicate that main hemodynamic parameters are obviously improved after stent implantation into cerebral aneurysm, and the blood velocity should never be neglectful in the treatment process.

Objective To evaluate the therapeutic effects and safety of open surgery vs.endovascular procedures for subclavian steal syndrome (SSS).Methods A retrospective analysis was made on 112 patients from July 2005 to July 2014 including 72 undergoing endovascular therapy and 40 cases treated by open bypass surgery.Results There were not difference between endovascular therapy group and surgical group in technical success rate (95.8％ vs.100％,P =0.239),postoperative subclavian artery diameter [(8.3 ± 1.6) mm vs.(8.1 ± 0.7) mm,P =0.374],postoperative suffering/contralateral systolic pressure ratio [(0.95 ±0.12) vs.(0.96 ±0.15),P =0.518].Nor there were the differences in mortality [0(0％) vs.3 (7.5％),P =0.018],postoperative renal dysfunction [2 (2.8％) vs.5 (12.5％),P =0.042],wound inffections [1 (1.4％) vs.4 (10％),P=0.034],duration of operation [(1.5 ±0.32) h vs.(2.7±0.51) h,P=0.027],days in hospital [(5.7±3) dvs.(9.3±3) d,P=0.013].After 1,3,5 years postoperative follow-up the two groups were not significantly different in patency rates,subclavian artery diameter,suffering/contralateral systolic pressure ratio (all P ＞ 0.05).Conclusion For subclavian artery steal syndrome,endovascular treatment is safe,effective and minimally invasive.

Objectives To investigate the effects of lipoxin receptor agonist BML-111 on IFN-βand IE86 mRNA expression of macrophages infected by human cytomegalovirus (HCMV). Methods Macrophages were infected with HCMV (MOI=0.5), and the cultured cells were randomly divided into control group, HCMV group, HCMV+BML-111 group, and HCMV+MP group. The cells were collected at 0,1,2,4,8 and 12 h after infection, and the levels of IFN-βand IE86 mRNA were tested by real-time PCR. Results Compared with HCMV group, the levels of IFN-βmRNA in HCMV+BML-111 group increased significantly (P < 0.05), while the levels of IFN-βmRNA in HCMV+MP group decreased significantly (P < 0.05); Compared with HCMV group, there were no significant differences of the levels of IE86 mRNA in HCMV+BML-111 group (P>0.05), while the levels of IE86 mRNA in HCMV+MP group increased significantly (P < 0.05). Conclusion BML-111 exerts antiviral activity by promoting the expression of IFN-βmRNA at the early stage of HCMV infection.

This paper aims to discuss the use and management of operation instrument in treatment of the wounded on the sea by using the new type of field operation instrument set so as to improve the rescue ability on the sea. The operation set has a new structure and is designed reasonably. It is easy to be washed and disinfected. It can be packed sterilizedly for a long time and deployed rapidly. All of the operation instrument is fixed in the box to prevent it from falling when it is used on the sea and it is convenient to count and arrange. The new type of field operation instrument set is fit for rescuing the wounded on the sea. It is convenient to be used, sterilized, stored and carried.

Aims HDAC2 gene was cloned into pEGFP-C2 vector to explore the efficiency of the plasmid trans-fection in renal fibroblasts COS-7 cells to identify the expression of both mRNA and protein levels and to ob-serve the distribution of the protein. Methods The HDAC2 cDNA was amlified by PCR and cut with the double enzyme Xho I and BamH I, then inserted into the eukaryotic expression vector pEGFP-C2 with T4 en-zyme. The recombinant vector was verified by PCR, restriction enzymes cut and sequencing identification. Then it was transfected into COS-7 cells and the ex-pression of pEGFP-C2-HDAC2 was monitored by fluo-
rescence microscope and PCR. Results Fragments of HDAC2 could be seen after dealt with double diges-tion, and GFP could also be detected in the transfected COS-7 cells. HDAC2 gene expression could be detec-ted by PCR and Western blot. The fusion expression of pEGFP-C2-HDAC2 could be detected by Western blot. Conclusion Eukaryotic expression vector of HDAC2 has been successfully constructed, the fusion expres-sion of HDAC2 and GFP protein can be detected in COS-7 cells.