Objective To establish analysis methods of two-dimensional(2-DE)gel electrophoresis for human osteosarcoma.Methods A series of methods,including Immobilized pH gradient were used as ID.Some applications,such as sample preparation used as choice of IPG gel,were improved.Coomassie brilliant blue staining,ImageMaster 2D Elite 3.01 analysis software,MALDI-TOF/ TOF MS and SWISS-PROT database serching were used to separate and indentify the proteins of human osteosarcoma.Results The good use pattern including repetitive experiments showed that in three experiments,the amount of protein spots of the same team sample deviates from the relative standard as following,the average of variation coefficient(%) : 23.00?10.11 and 20.33?9.90;and the range of variation coefficient(%) were:3.80-6.89 and 2.706.89 from osteosarcoma and normal group respectively.The isoelectric point and molecular weigh of the same protein spots in three experiments deviated from relative standard as following:(8.93%?1.17)%,(10.16?2.02)%,(10.87?3.86)%,respectively.Therefore,better resolution and repetitive 2-DE atlas were obtained.The proteins from 11 pairs of sample were analysed by mass spectrometry,9 identified proteins(transthyretin precursor, Triosephosphate isomerase,slow skeletal muscle,cardiac muscle Troponin T,Cofilin-1,Myosin light chain 1,Calgranulin B,Heat-shock protein,Annexin A5, Fanconi anemia group D2 proteins) were more abundant in osteosarcoma tisstues and 2 proteins manganese SOD and carbonic dehydratase appeared down-regulation in osteosarcoma tisstues.Conclusion This optimized 2-DE map is an important tool for further study on osteosarcoma,and these identified proteins were related-proteins with osteosarcoma.It is suggested that the changes of the proteome are involved in the pathology processe of osteosarcoma.

Objective To investigate the method of establishing a model of delay eyeblink condi-tioning ( DEC) in no surgical cynomolgus monkey and analyze the related acquisition rule.Methods Using the special monkey chairs to fix 6 adult male cynomolgus monkeys,they were sitting on the chairs and keep-ing awake,and heads could rotate freely during the whole training session.In the experiment,the 6 monkeys were trained in DEC with a tone (500 ms,85 dB) as the conditioned stimulus (CS),paired with a corneal oxygen-puff (100 ms,5 psi) as the unconditioned stimulus (US).There were 120 trials per-session,and 2 sessions per-day.During the experiments,an infrared emitter/detector attached to spectacles for eyeblink re-cording,the data of conditioned response ( CR) and startle response ( SR) were analyzed offline.Result-s Among the 6 cynomolgus monkeys,4 of them completed all of the training process of DEC and 3 monkeys had successful acquisition of DEC.The average CR rate in the last training session reached ( 64.67 ± 2.00)%,(P<0.01);1 monkey only showed a high acquisition rate( 85.00%) at the first training session, and the low CR rate in the next training sessions with the average rate of 18.16%,suggesting that the DEC model was failed to established.The SR rates in 4 cynomolgus monkeys were low with average rate of 5.47%. Conclusion Although the DEC behavior training of the cynomolgus monkey is easy to be influenced by many inside or outside factors,DEC can be obtained under the condition of non surgery and proper braking.

Objective:To compare the postoperative effects of double eyelid surgery with different exophthalmos to find its influence on the surgery and necessary changes in preoperative design and during operation.Methods:A total of 50 female patients with single eyelid seeking beauty from June 2021 to March 2022 were selected from the Department of Plastic Surgery, Affiliated Hospital of Qingdao University. The ocular protrusion was measured by HETEL ophthalmostatometer before surgery. Both eyes at 12-15 mm were taken as normal group ( n=26), both eyes at 16-18 mm as mild protrusion group ( n=14) and both eyes at 19-22 mm as severe protrusion group ( n=10). All the patients were treated with double-eyelid surgery by orbital septum and unified postoperative nursing. Results:After six months of follow-up, there was no difference in eyelid width with closed eyes (all P>0.05). The width of double eyelid with open eyes in normal group was smaller than that in mild protrusion group ( F=23.23, all P<0.05), and the width of double eyelid with open eyes in mild protrusion group was smaller than that in severe protrusion group ( F=47.70, all P<0.05). There was no difference in the improvement rate of facial aesthetics among the three groups ( P>0.05). The " feeling of meet" and scar formation in the normal group were less than those in the mild protrusion group ( F=16.92, F=33.45, all P<0.05), and the " feeling of meet" and scar formation in the mild protrusion group were less than those in the severe protrusion group ( F=27.93, F=28.53, all P<0.05). The improvement rate of normal group was higher than that of mild and severe protrusion group (χ 2=7.25, 7.89, all P<0.05). There was no difference in the improvement rate between the mild and severe protrusion groups ( P>0.05). Conclusions:In clinical practice, it is necessary to make corresponding changes in the preoperative design and operation of double eyelid surgery for patients with high eyeball protrusion.