Objective To construct the typeⅢ secretion system (T3SS) deficient mutant of O157∶H7 EDL933, and to detect its ability of attachment after infection with HeLa cells.Methods The recombinant DNA fragments obtained kana-mycin resistant gene were constructed by overlap extension PCR and transferred into EDL933/pKD46 to replace escR gene by homologous recombination.Results The T3SS-deficient mutant (ΔescR) was successfully construted.Compared with the wild-type EDL933, the growth rate and attachment on HeLa cells of ΔescR were significantly decreased.Conclusion The deletion of escR gene, which encodes T3SS structural protein EscR, affects the attachment of EDL933, suggesting a better basis for future studies of T3SS.

Objective To construct a prokaryotic plasmid expressing the recombinant protein of enterohemorrhagic Escherichia coli(EHEC) effector NleB1 and to prepare the polyclonal antibody of mouse anti-NleB1.Methods The nleB1 (990 bp) gene was amplified from the genome EHEC O157∶H7 and cloned into the expression plasmid pET24a to construct the recombinant plasmid pET24a-nleB1 that was transformed into E.coli BL21(DE3).After induction with isopropylthio-gelactoside( IPTG) , the His-tag fusion proteins were purified by Ni+affinity chromatography and gel slices.The polyclonal antibody was prepared by immunizing BALB/c mice with purified recombinant proteins and analyzed by Western blotting and ELISA.Results The pET24a-nleB1 recombinant plasmid was successfully constructed, the fusion protein was ex-pressed and purified,and the polyclonal antibody was obtained by immunizing mice with purified fusion protein.Western blotting and ELISA staining demonstrated that the polyclonal antibody was successfully obtained.Conclusion The prepara-tion of the polyclonal antibody against EHEC O157∶H7 NleB1 will be of help for further studies on the function of NleB1 protein.

OBJECTIVETo provide more proofs for expounding the genetic relationships among the (varietal) species in genus Fritillaria from Sichuan province.

METHODThe ISSR marker technique was used to study relationships and genetic polymorphism of nineteen populations in ten species and one varietal species of genus Fritillaria. Genetic similarities were calculated by using NTSYS software and the dendrogram was constructed by using UPGMA method.

RESULTEleven primers were selected from 35 ISSR primers, and 179 DNA fragments were amplified from 19 populations. Of which, 179 fragments were polymorphic (percentage of polymorphic bands was 86.8%). The genetic similarity among all accessions ranged from 0.569 to 0.855. Clustering analysis showed that the 19 populations of Fritillaria could be distinctively classified into 4 groups. F. cirrhosa, F. przewalskii, F. cirrhosa var. logirnectarea and F. dajitensis were in the first group; The second group was the cluster of F. cirrhosa and F. mellea (wild and cultivated species); The third group was F. sulcisquamosa, F. thunbergii, wabunesis and F. delavayi; F. hupehensis alone formed the fourth group.

CONCLUSIONISSR marker technique is suitable for the genetic diversity of Fritillaria from Sichuan province. Interspecific identifications among the four original species of Bulhus Fritillariae Cirrhosae recorded by pharmacopoeia of China, and between them and the other species of genus Fritillaria from Sichuan province could not be gained by using ISSR markers technique. In addition, the cluster result of genus Fritillaria had some relationships with the geographical distribution.

China ; Fritillaria ; classification ; genetics ; Genetic Variation ; Microsatellite Repeats ; Phylogeny

Complex surgeries often need multi-disciplinary surgeons to work collaboratively and set synthesized operation plans. Aided by collaboration enabled three dimensional visualization software over network, operation plans can be made more intuitively and accurately. Because different disciplinary surgeons often work at different locations, a distributed collaboration virtue environment over network should be provided. By our knowledge, such a system does not exist yet. Toward this, we implement a platform upon Client/Server architecture over network. The 3D model is reconstructed from CT image data of DICOM format and the resulting mesh is then simplified, using the Visualization Toolkit (VTK). The simplified polygon mesh data is further seamlessly integrated into our 3D graphics system developed with the HOOPS/3DAF for displaying, where the model is converted to the lossless compression stream file format-HSF, which is suited for network transmission. Using this format, a collaboration enabled interactive visualization system is implemented upon Client/Server architecture, developed with HOOPS/NET toolkit. The reconstructed 3D model is clear, and the interactive collaboration with 3D visualization is fairly real-time. We then implemented a platform for developing collaborative surgery simulation software, into which the surgery operation simulation and prosthesis design function can be easily added.
Computer Simulation ; Diagnostic Imaging ; methods ; Humans ; Image Enhancement ; methods ; Image Interpretation, Computer-Assisted ; methods ; Imaging, Three-Dimensional ; methods ; Internet ; Software ; User-Computer Interface

Having manufactured the bone fatigue damage testing device SL-2000, we applied it to research on bone fatigue damage and on the bone microdamage which can indicate bone biomechanical property. According to the general principle of industry fatigue machine and bone fatigue test, the strength, frequency and times of loading, temperature and degree of wetness were controlled in the SL-2000 Device for making the experiment condition stable. After the fatigue damage to SD rats' vertebrae, the models of bone microdamage in SD rat were established. 0-200 N of strength, 0-10 Hz frequency, and 0-999999 times of loading, ambient temperature 50 degrees C experiment, and approximately 99% of the ambient degree of wetness could be adjusted continuously. Three kinds of microdamage such as microcrack, cross-hatch staining, and diffuse staining were observed in the SD rats' vertebrae fatigue damaged by the device. The microcrack density was 19.76+/-15.05 #/mm2, the microcrack length was 36. 74+/-11. 51 microm, and the area ratio of diffuse staining was 0.4117%. Therefore the device is suitable for bone fatigue damage test and for establishment of the model of bone microdamage.
Animals ; Biomechanical Phenomena ; Compressive Strength ; Equipment Design ; Fractures, Stress ; diagnosis ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Spinal Fractures ; diagnosis ; physiopathology ; Spine ; pathology ; Weight-Bearing

Objective To investigate whether the preoperative nutritional risk index(NRI) can predict the clinical out-comes of the patients underwent homologous orthotopic heart transplantation after 4 weeks.Methods With the use of continu-ous sampling method, a total of 168 adult patients who underwent homologous orthotopic heart transplantation from March 2013 to March 2015 were enrolled in this study.According to the cut-off points of the NRI, the patients were divided into 4 groups (high, moderate, mild and no nutritional risk).4 weeks after transplantation, the data about the length of stay in ICU, infec-tive and non-infective complications were collected , and the predictive value of the preoperative NRI for the clinical outcomes of the patients underwent homologous orthotopic heart transplantation were evaluated after 4 weeks of operation.In addition, the enrolled patients were divided into 2 groups according to their nutritional risk(with or without), and the differences between the two groups were compared.Results Among the patients, the prevalence of high, moderate, and low nutritional risk was 7. 1％, 32.7％ and 9.5％, respectively.The prevalence of infective complications for the 4 groups(from high risk to no risk) was 16.7％、7.3％、6.3％ and 3.5％, and the corresponding prevalence of non-infective complications was 16.7％、9.1％、6.3％ and 2.4％, respectively.In addition, the length of ICU stay of the 4 group was(10.9 ±1.0)days,(7.8 ±0.5)days, (6.7 ±0.7)days and(5.9 ±0.6)days, respectively.There were significant differences of length of ICU stay, infective and non-infective complications among the 4 groups(P<0.05 for all).Totally, there were significant differences of clinical out-comes between patients with and without nutritional risk(P <0.05 for all).Conclusion Preoperative NRI can predict the clinical outcomes regarding infective and no-infective complications as well as length of ICU stay for the adult patients under-went homologous orthotopic heart transplantation .

OBJECTIVETo assess the efficacy and experience of gracilis muscle transposition for complex rectovaginal fistula (RVF) and rectourethral fistula (RUF).

METHODSNineteen patients underwent gracilis muscle transposition for complex RVF and RUF from May 2009 to November 2011 in the Beijing Shijitan Hospital and the clinical data were prospectively collected. The success rate and complications were recorded. SF-36 quality of life score, Wexner fecal incontinence score, and female sexual function score before surgery and 6 months after surgery were recorded.

RESULTSIn 19 patients, there were 8 males (RUF) and 11 females (RUF). The times of failed attempt repair preoperatively ranged from 0-3 (mean, 1.0). The diameter of the fistula ranged from 0.5-3.0 cm (mean, 1.6), and all fistulas located above the sphincter. The operative time ranged from 145-400 minutes (median, 240). The postoperative hospital stay ranged from 10-39 days (median 21). Early postoperative complications included thigh pain and numbness in 2 cases, leg numbness in 2 cases. No long-term complications were noticed. The follow-up period ranged from 6-35 months (median, 18). The gracilis muscle transposition had a healing rate of 94.7% (18/19). As compared with the preoperative level, Wexner score decreased from 10.0±8.8 to 2.9±5.8, and the continence function improved significantly (P=0.002). Sexual function score of 11 female patients increased from 1.0±1.8 to 4.0±4.0, and the sexual function had a significant improvement after surgery (P=0.022). SF-36 quality of life scores improved significantly (P<0.001).

CONCLUSIONSGracilis muscle transposition for complex rectovaginal fistula and rectourethral fistula has high success rate with mild and rare complications.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Muscle, Skeletal ; surgery ; Prospective Studies ; Rectal Fistula ; surgery ; Rectovaginal Fistula ; surgery ; Surgical Flaps ; Thigh ; surgery ; Treatment Outcome ; Urethra ; surgery ; Urinary Fistula ; surgery ; Young Adult

In order to characterize the effects of 17β-estradiol (17β-E2) on the expression of IL-6,IL-11 and NF-κB in the human MG-63 osteoblast-likecell line, the expression of IL-6 was detected by RT-PCR, Northern blot and Western blot. The expression of IL-11 was determined by RT-PCR, and NF-κB by Western blot. The results showed that 17β-E2 down-regulated the expression of IL-6 mRNA and protein, IL-11 mRNA and NF-κB protein in MG-63 cells. It was suggested that the expression of NF-κB, IL-6 and IL-11 in MG-63 cells could be suppressed by 17β-E2, and this might lend support to estrogen replacement therapy in postmenopausal women.

OBJECTIVETo evaluate the sensitivity and specificity of CD8CD28/CD8CD28T lymphocyte balance in predicting the gastrointestinal hemorrhage (GH) in patients with inflammatory bowel disease (IBD).

METHODSForty-nine IBD patients, including 30 with ulcerous colitis (UC) and 19 with Crohn's disease (CD), were enrolled to test peripheral blood CD8CD28and CD8CD28T cells using flow cytometry. All the patients were followed up for one year. The receiver-operating characteristic (ROC) curves were used to test the efficiency of CD8CD28/CD8CD28T lymphocyte balance to predict GH. The differences in lasting time of remission (LTR) under different factors were compared using Kaplan-Meier survival analysis, and the correlation between CD8T lymphocytes and the factors were analyzed.

RESULTSThe utilization rates of immunosuppressant, steroids, and biological agent (BA) were significantly higher in CD patients than in UC patients (P=0.003, 0.043 and 0.002, respectively). The frequencies of CD8CD28T cells were obviously higher in UC patients than those in CD patients (t=3.022, P=0.004). CD8CD28T cells, CD8CD28T cells, and especially CD8CD28/CD8CD28ratio (area under curve of 0.977, P=0.000; cut-off value of 1.14 [13.95%/12.24%] with a sensitivity of 93.3% and a specificity of 91.2%) showed good efficiencies in predicting GH (P<0.01). The mean and median of LTR of IBD patients who did not receive BA or surgical treatment were significantly longer (Χ=9.730, P=0.002; Χ=15.981, P=0.000). CD8CD28/CD8CD28ratio was significantly related to both BA (P=0.009) and surgery (P=0.038).

CONCLUSIONBoth decreased CD8CD28T cells and elevated CD8CD28T cells are closely correlated with GH, and their ratio can predict the occurrence of GH with a high sensitivity and specificity and is correlated with BA and surgery at the cut-off value of 1.14.

To analyze the use of outcome indicators of randomized controlled trial(RCT) of acupuncture in the treatment of acute ischemic stroke in recent three years, so as to provide a basis for building a study on the core outcome indicators for the treatment of acute ischemic stroke with acupuncture. The RCTs of acupuncture treatment for acute ischemic stroke in recent three years were collec-ted through computer retrieval of eight Chinese and English databases and two clinical trial registries at home and abroad. Literature was screened out, and data was extracted. Risk of assessment bias tool Cochrane 6.1 was used for bias risk assessment, outcome indicators were summarized and analyzed. A total of 47 RCTs were included, and 3 studies were trials registration scheme. Outcome indicators were divided into 6 categories according to functional attributes, namely physical symptoms/signs, physical and chemical examination, quality of life, traditional Chinese medicine symptoms/syndromes, safety events and long-term prognosis. The study found that in addition to the common problems in previous studies covered by the status quo of outcome indicators selection of RCT of acupuncture in the treatment of acute ischemic stroke, there were also the other problems as follows: emphasis on macroscopic efficacy indicators but neglect of acupuncture specific indicators, lack of characteristic indicators and economic indicators of traditional Chinese medicine therapy, and unification of indicators measurement tool and measurement time point. In the future, the outcome indicators set for the treatment of acute ischemic stroke with acupuncture shall be established, and the core outcome indicators set shall be in line with the characteristics of traditional Chinese medicine treatment.
Acupuncture Therapy ; Brain Ischemia/therapy* ; Humans ; Ischemic Stroke ; Quality of Life ; Randomized Controlled Trials as Topic ; Stroke/therapy* ; Treatment Outcome