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AIM: To investigate the effects of TNF receptor-associated death domains (TRADD) of latent membrane protein 1 (LMP1) on the proliferation of nasopharyngeal cancer SP18 cells.METHODS: The SP18-LMP1 cells and SP18-LMP1TRADD cells, which expressed LMP1 and LMP1 TRADD proteins, respectively, were established.The proliferation of SP18 cells affected by LMP1TRADD was detected by cell counting to analyze the cell growth curve, and by colony formation assay, soft agar formation assay, and flow cytometry.Moreover, the expression profile of differential genes between SP18-LMP1 cells and SP18-LMP1TRADD cells was analyzed by gene chips.RESULTS: The cell growth curve, and the results of colony formation and soft agar formation displayed that the growth velocity and colony forming ability of SP18-LMP1 cells were stronger than those of SP18-LMP1TRADD cells (P<0.01).The results of flow cytometry analysis showed that the proliferation index of SP18-LMP1 cells was higher than that of SP18-LMP1TRADD cells (P<0.01).Sixty-three differentially expressed genes associated with cell proliferation were screened out, in which 33 genes were up-regulated and 30 were down-regulated in the SP18-LMP1TRADD cells.CONCLUSION: TRADD active region is an important functional site of LMP1 to promote the proliferation of SP18 cells.LMP1 may improve the cell proliferation index and induce the proliferation of SP18 cells through TRADD.
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Objective To compare the changes of serum levels of heme oxygenase‐1(HO‐1) ,C‐reactive protein(CRP) and Chitinase‐3‐1ike protein 1(YKL‐40) in patients with rectal cancer after laparoscopic‐assisted radical operation versus open radical surgery .Methods According the integration and elimination standard ,60 cases of patients with rectal cancer were selected and di‐vided into laparoscopic group (30 cases) and open group (30 cases) ,the concentrations of HO‐1 ,YKL‐40 and CRP in peripheral blood of patients were detected on the first and third postoperative day and preoperative day(P<0 .05) ,and were compared between the two groups .Results The serum levels of HO‐1 ,YKL‐40 and CRP in the laparoscopic group were significantly lower than those in the open group on the first and third postoperative day(P<0 .05) .The serum levels of HO‐1 ,YKL‐40 and CRP were significant‐ly increased after surgery ,and serum levels of HO‐1 and CRP on third postoperative day were significantly increased compared with that on the first postoperative day in the two groups after surgery ,while serum level of YKL‐40 on the third postoperative day was significantly lower than that on the first postoperative day in the two groups after surgery .Conclusion The stress reaction is com‐paratively mild in patients with rectal cancer after laparoscopic assisted radical operation ,and the postoperative period may be shor‐ter in patients treated with laparoscopic assisted radical operation than those treated with open radical surgery .
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To understand the Escherichia coli (E .coli) O157∶ H7 isolated from cow in Zhengzhou ,Henan Province ,a total of 146 samples of cow fecal and milk were collected in the different farms ,and E .coli O157∶ H7 was detected with mul-tiplex polymerase chain reaction (PCR) in our laboratory .Then the biochemical characteristics ,growth dynamic ,the biofilm formation ,and the toxin genes of the E .coli O157∶ H7 isolates were analyzed .The results showed that 2 strains of E .coli O157∶H7 were found ,with the detection rate of 1 .4% ,and the isolates were named as L1 and L2 in current study ,respec-tively .The E .coli O157∶H7 clinical isolates had the same biochemical characteristics with that of the typical E .coli .The L1 and L2 isolates presented similar growth curve ,which entered into the log phase earlier than that of the standard strain .L1 strain formed thick ,confluent ,complete biofilm after 48 hours post-inoculation ,and the biofilm of L2 strain was formed com-pletely in 36 hours .The two E .coli O157∶ H7 isolates were positive with eaeA and hlyA genes ,and the L1 strain also carried the Stx2 virulence gene .Our results reinforce the epidemiological data of E .coli O157∶H7 ,and underscore the need for more effective surveillance of animal-derived E .coli O157∶H7 isolates in Zhengzhou City ,China .
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To improve the expression level of tmAMP1m gene from Tenebrio molitor in Escherichia coli, we studied the effects of expression level and activity of the fusion protein HIS-TmAMP1m by conditions, such as culture temperature, inducing time and the final concentration of inductor Isopropyl beta-D-thiogalactopyranoside (IPTG). We analyzed the optimum expression conditions by Tricine-SDS-PAGE electrophoresis, meanwhile, detected its antibacterial activity by using agarose cavity diffusion method. The results suggest that when inducing the recombinant plasmid with a final IPTG concentration of 0.1 mmol/L at 37 degrees C for 4 h, there was the highest expression level of fusion protein HIS-TmAMP1m in Escherichia coli. Under these conditions, the expression of fusion protein accounted for 40% of the total cell lysate with the best antibacterial activity. We purified the fusion protein HIS-TmAMPlm with nickel-nitrilotriacetic acid (Ni-NTA) metal-affinity chromatography matrices. Western blotting analysis indicates that the His monoclonal antibody could be specifically bound to fusion protein HIS-TmAMPlm. After expression by inducing, the fusion protein could inhibit the growth of host cell transformed by pET30a-tmAMP1m. The fusion protein HIS-TmAMP1m had better stability and remained higher antibacterial activities when incubated at 100 degrees C for 10 h, repeated freeze thawing at -20 degrees C, dissolved in strong acid and alkali, or treated by organic solvents and protease. Moreover, the minimum inhibitory concentration results demonstrated that the fusion protein HIS-TmAMP1m has a good antibacterial activity against Staphylococcus aureus, Staphylococcus sp., Corynebacterium glutamicum, Bacillus thuringiensis, Corynebacterium sp. This study laid the foundation to promote the application of insect antimicrobial peptides and further research.
Subject(s)
Animals , Anti-Infective Agents , Pharmacology , Antimicrobial Cationic Peptides , Genetics , Pharmacology , Escherichia coli , Genetics , Metabolism , Genetic Vectors , Genetics , Insect Proteins , Genetics , Recombinant Fusion Proteins , Genetics , Pharmacology , Tenebrio , ChemistryABSTRACT
This paper presents a new method of separating the junction between the epidermis and the dermis with enzyme,and then,observing the small wound of the papillary of the dermis by means of SEM.The result showed that this new method would be advantageous to the identification of small stab wounds and providing some concrete identify targets.
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Han Jiangbo. Department of Theoretical Mechanics, Luoyang Industry College. Luoyang Since January 1982 we have relocated the insertion of a part of pectoralis major or deltoid muscle in the treatment of 5 cases of habitual dislocation of shoulder. After over a year of follow-up no recurrence of dislocation has been found in all the cases. The shoulder joint seemed to be stable, the range of movements increased, and disturbance in supination was corrected. The operative procedure was studied in fresh cadavers and human skeleton. Studies on mechanical model of shoulder joint, with theoretical calculation of changes in pronation after muscle displacement, showed that movements of pronation were remarkably increased after displacement of partial pectoralis major and deltoideus. It could also compensate the weatened subscapularis.