3. Construction and identification of a vector expressing short hairpin RNA targeting annexin II gene
Academic Journal of Second Military Medical University 2006;27(11):1181-1185
Objective: To construct a vector encoding short hairpin RNA(shRNA) targeting annexin H and to observe its influence on annexin II expression. Methods: Annexin II-targeted hairpin RNA1 and RNA2 were devised according to the Gen-Bank annexin II sequence. Meanwhile, a nonspecific sequence was taken as negative control. The oligonucleotide strands of DNA fragments encoding the above shRNAs were synthesized. After annealing of the complementary strands, the DNA fragments were cloned into human pGenesil1 plasmid followed by amplification in E. coli and sequencing. The 3 pGenesil1 constructs were then transfected into HEK293 cells and the expression of annexin II was examined by real-time fluorescence quantitative RT-PCR and Western bolt. The non-transfected cells were taken as blank control. Results: Restrictive enzyme digestion and sequencing verified that the 3 recombinants (pGenesil1-annexin II shRNA1, pGenesil1-annexin II shRNA2, and pGenesil1-negative shRNA) were correct. The annexin II expression was significantly lower in HEK293 cells transfected with pGenesil1-annexin II shRNA2 compared with those transfected with pGenesil1-annexin II shRNA1, pGenesil1-negative shRNA, and blank control (P<0.05); and there was no significant difference between the latter 3. Conclusion: We have successfully generated an annexin II-targeted shRNA construct, which can significantly inhibit the expression of annexin II.
4. Establishment and optimization of two-dimensional gel electrophoresis for separation of human kidney phosphoproteome
Academic Journal of Second Military Medical University 2006;27(6):603-606
Objective: To separate human kidney phosphoproteome by two-dimensional gel electrophoresis. Methods: The phosphorylated proteins from human kidney tissues were enriched with phosphate metal affinity chromatography (PMAC) resin. After being concentrated and desalted, the samples were separated by isoelectric focusing on first dimension and SDS electrophoresis on second dimension. Results: The phosphorylated proteins were successfull y extracted from human kidney tissues and were separated by two-dimensional gel electrophoresis. Conclusion: Phosphoprotein enrichment technique combin ed with two-dimensional gel electrophoresis is an effective approach to study phosphoproteome, laying a foundation for further investigation of human kidney phosphoproteins.
5.Application of multi-slice spiral CT in diagnosis of tracheobronchial foreign bodies in children
Jianqiang WEI ; Ya ZHANG ; Bo LIU
Journal of Practical Radiology 2014;(11):1879-1882
Objective To evaluate the clinical value of multi-slice spiral CT in diagnosing tracheobronchial foreign bodies in chil-dren.Methods 139 patients with airway foreign body confirmed by bronchoscopy were examined with 128-slice spiral CT,and then the CT data was reconstructed by using multiple-planar reconstruction(MPR),curved planar reconstruction (CPR),volume render-ing(VR),minimum intensity projection(MinIP)and virtual endoscopy (VE).The CT results were compared with bronchofibersco-py findings.Results Among the 139 cases,the foreign bodies located at trachea in 13 cases,right bronchia in 75 cases,left bronchia in 48 cases,and bilateral bronchia in 3 cases.137 cases (98.6%)were correctly diagnosed,and 2 cases (1.4%)were missed on CT.Although the difference of the results at trachea and right main stem bronchus was statistically significant (P <0.05),the over-all results was not significant difference between CT and bronchoscopy examination (P =0.340).Associated findings included em-physema in 74 cases,pneumonia in 48 cases,atelectasis in 22 cases,mediastinal displacement in 57 cases,lung interstitial emphyse-ma in 4 cases,pneumomediastinum and pneumoderma in 4 cases,bronchiectasis in 2 cases,and pleural effusion in 4 cases.Conclu-sion MSCT and its postprocessing techniques can accurately and intuitively display the tracheobronchial foreign body and complica-tions,which has important value for clinical application.
8. Systhesis of magnetic thermosensitive hydrogel and its heat effect under alternating magnetic field
Chinese Pharmaceutical Journal 2012;47(9):693-695
OBJECTIVE: To synthesize magnetic thermosensitive hydrogel and study its heat effect under alternating magnetic field in vitro. METHODS: PLGA-PEG-PLGA triblock copolymer was synthesized by ring-opening polymerization of D, L-lactide and glycolide with PEG1500 in the presence of stannous iso caprylate. Magnetic thermosensitive hydrogel of different concentrations were prepared using different currents. The influences of the concentration of magnetic fluid and current of magnetic field on the heat effect were separately observed. RESULTS: The synthetized PLGA-PEG-PLGA triblock copolymer was excellently temperature sensitive. It retained the thermo-sensitivity of original hydrogel when the magnetic fluid was loaded. The 5, 10 and 20 min heating ability of magnetic fluid was positively linearly correlated with its concentration (r=0.9985, 0.9893 and 0.9711, respectively, n=3) and current of magnetic field (r=0.9948, 0.9977 and 0.9994, respectively, n=4). CONCLUSION: The magnetic thermosensitive hydrogel has excellent temperature sensitivity. When alternating magnetic field is applied, the temperature of the system can rise and reach above LCST of hydrogel. The temperature can be controlled by changing the concentration of magnetic fluid and the current of magnetic field. Copyright 2012 by the Chinese Pharmaceutical Association.
9. Limb characteristics of Yangshao residents in Wanggou site in Zhengzhou
Acta Anatomica Sinica 2020;51(1):114-123
Objective To research the physical development of ancient inhabitants in the Neolithic Age. Methods Referring to Anthropometric Method , the main projects including humerus, tibia and femur of 108 human bone specimens unearthed from Wanggou site have been measured respectively. The limb bone index, sexual dimorphism and height have been compared and analyzed by Excel software. Results The overall development level of humerus and femur was higher than that of the control group in the same period and weaker than that of the Inner Mongolian group. The development of the middle femur was weaker than that of the control groups, with less muscle attachment. The difference in the brawniness of humerus between the genders was statistically significant (i = 0. 811, P>0. 05) , while the difference in the brawniness of femur was just the opposite (t = 0. 63, P>0. 05 ). The maximum variation range of the male femur was 40. 6 to 49. 4 cm, which was at a moderate low level in the control group. The average height of male was 166. 22 cm, ranging from 164. 47-167.68 cm. The average height of female was 158.06 cm, ranging from 154.02-160.34 cm; There was a significant difference in height between the genders (i = 4. 96, P<0. 01).The height of male is close to the south China group, lower than the comparison group in the same region and the north region, and that of female is at a higher level in the south region. The sexual dimorphism index of Wanggou formation was 5. 7 %. Conclusion The humerus, tibia and femur of the male in Wanggou group are stronger than that of the female, while the upper limbs of the male were stronger than that of the female, and there was no significant difference in the strength of the lower limbs. We suggest that there is no significant difference in nutrition between male and female of Wanggou site during their growth.