1.Progress in Aptazymes
Progress in Biochemistry and Biophysics 2006;0(06):-
Aptazymes are a new artificial synzyme, selected from the random oligonucleotide sequence libraries against various of effector molecules. They own the advantages of an aptamer(the receptor site) and the ribozyme (the catalytic active site). Moreover, aptazymes as catalytic molecular beacon provide a new orientation for the quantitative analysis of effector molecules. Aptazymes not only have the application in genomics and proteomics, but also have potential applications in biosensor and DNA AND gate.
2.The application of clients' relationship management in large general hospital physical examination centre
Lan TANG ; Yuanxian ZHANG ; Fanling ZENG
Chinese Journal of Medical Education Research 2011;10(8):986-988
With the economic development, more and more people concerned about health. Institution and people engaged in physical examination is springing up. Facing increasingly less obvious advantages of product homogeneity, and the growing market which must cater to different customers, as a large general hospital medical center, we should make good management of customer relationship, improve customers' satisfaction, increase their loyalty, provide the most effective health protection for the physical examination so as to improve market competitiveness and bring social and economic benefits to the hospitals, thus achieving a win-win objective.
3.Effects of Environmental Tobacco Smoke on Serum Lipid and the Lipases of Serum and Liver and Lung in Rats
Ming ZENG ; Ting HUANG ; Lan GUAN
Journal of Environment and Health 1992;0(04):-
0.05).The smoking group had a higher TG level and a lower HDL-C level than the control one with statistical significance(P0.05).The HL activities in serum,lung and liver of smoking group were lower than those of control group(P
5.Rapid purification of human albumin with immunomagnetic microsphere technology
Minghui WU ; Lingli JIANG ; Fanbo ZENG ; Nidan WANG ; Lan TANG
Acta Pharmaceutica Sinica 2006;41(7):608-614
Aim To develop a rapid and inexpensive method for purification of human albumin, a method of immunomagnetic microspheres (IMMS) based on enzyme-linked immunosorbent assay (ELISA)for the purification of human albumin from human serum. Methods Polystyrene magnetic microspheres with carboxyl groups as carriers were prepared, and then the carboxyl groups on the surface of the microspheres were activated by ethylcarbodiimide (EDC). Finally rabbit anti-human serum albumin (HSA) antibodies were covalently bound to it and the complex can specifically capture HSA. After the procedure of capturing HSA, through taking rabbit anti-human albumin protein antibodies as a capture antibody, and goat anti-human albumin protein antibodies as a detection antibody, an ELISA on IMMS was developed, which can determine the recovery yield of HSA from the human serum. Results The result of the experiment was that the recovery of human albumin with IMMS was (86 ± 4) % , and IMMS were reused for two other purifying cycles, the results of which were (69.0 ± 0.6) % and (40.8 ± 0.8) % , and the purity of the product was about 90%. Conclusion The results above prove that the immunomagnetic purifiying strategy was shown to be efficient and offers an new thought for a large scale production of highpurity HSA.
6.Subtle mutation analysis of survival motor neuron gene in families with spinal muscular atrophy
Jian ZENG ; Yanhong LIN ; Aizhen YAN ; Fenghua LAN
Chinese Journal of Laboratory Medicine 2012;35(7):607-611
ObjectiveTo establish a analytical system for the survival motor neuron (SMN) subtle mutation,and evaluate its application in two families with spinal muscular atrophy (SMA).MethodsSMN genes in seven family members from two SMA families were analyzed at both transcript level and genomic level,by the use of the conventional PCR-RFLP,allele-specific PCR,multiplex ligation-dependent probe amplification (MLPA) and T subcloning and sequencing of SMNI gene.ResultsIn family A,the patient had a single SMN1 copy who was carrying nonsense mutation L228X,which was also found in his father.In family B,as the patient's sample was unavailable,the father was indeed a carrier with one normal SMN1 allele and the other SMN1 allele carrying a frameshift mutation 22_23insA.The remaining family members were SMA carriers with one SMN1 copy.ConclusionThis analytical system for SMN subtle mutation offers viable molecular basis for genetic counseling and prenatal diagnosis in SMA families.
7.Establishment of ovarian SKOV3 cell line stably expressing PES1 by Tet-on inducible system
Jieping LI ; Qinren ZHUANG ; Xiaopeng LAN ; Guobin ZENG ; Xiaofeng LUO
Journal of International Oncology 2012;39(6):465-468
Objective To further research the biological functions of PES1,the ovarian SKOV3 cell line with inducible stable PES1 expression is established by using Tet-on system.Methods PES1 was cloned into pTRE-Tight vector via PCR and its expression was identified. After transfected the regulating plasmid pTet-on,SKOV3 cells were screened with G418 and re-transfected pTRE-Tight-PES1.The positive cell clones were screened out with hygromycin and were induced by doxycycline (Dox) to definite the best induction concentration.Growth velocity of SKOV3 cells stably expressing PES1 induced by Dox was detected with viola crystallina.Results The SKOV3 cells with inducible PES1 expression were screened out after the cells were transfected pTRE-Tight-PES1 constructed.Dox could dose-dependently induce the PES1 expression with the concentration under 2 mg/L,and 2 mg/L of Dox induced the highest PES1 expression.Growth velocity of SKOV3 cells transfected pTRE-Tight has no significant difference between the SKOV3 cells transfected nothing induced with Dox.However,the SKOV3 cells transfected pTRE-Tight-PES1 grew faster than the cells transfected pTRE-Tight or without transfection in the fourth day (P =0.001 ).Conclusion The inducible stable PES1 expression SKOV3 cells are successfully established and could be used to be an effective cell model to research the biological functions of PES1.The expression of PES1 could promote the growth of SKOV3 cells.
8.Prokaryotic Expression and Purification of HIV-1 Vif and hAPOBEC3G, Preparation of Polyclonal Antibodies
Lan LI ; Yishu YANG ; Zelin LI ; Yi ZENG
Virologica Sinica 2008;23(3):173-182
To prepare HIV-1 Vif and hAPOBEC3G and to produce their antibodies, the full length gene fragment of HIV-1 Vif was amplified by PCR from a plasmid of HIV-1 NL4.3 cDNA, and the APOBEC3G gene was obtained by RT-PCR from the total RNA of H9 cells. The resulting DNA construct was cloned into a prokaryotic expression vector (pET-32a). Recombinant pET-vif and pET-APOBEC3G were expressed respectively in Eserichia coli BL21 (DE3) as an insoluble protein. The vector also contained a six-histidine tag at the C-terminus for convenient purification and detection. To express and purify the HIV-1 Vif and hAPOBEC3G in E. coli cells, the accuracy of inserted gene and specificity of proteins were detected by the two enzyme digestion method, SDS-PAGE, and Western blotting. Rabbits were then immunized by Vif or APOBEC3G protein and serum samples were tested by indirect ELISA to determine the level of antibodies. Immunoenzyme and immunofluorescence assays were performed to identify the specificity of polyclonal antibodies. The titer of the anti-Vif antibodies was 1:204800, and that of the anti-APOBEC3G antibodies was 1:102400. Thus the antibodies could detect the antigen expression in the cells, demonstrating that fusion proteins with high purity and their corresponding polyclonal antibodies with high titer and specificity were achieved.
9.Accelerated solvent extraction of ecdysterone from Achyranthes bidentata
Jiaquan WANG ; Minjun ZHAO ; Lan LIN ; Yinglei CHAI ; Su ZENG
Chinese Traditional and Herbal Drugs 1994;0(12):-
Objective To apply accelerated solvent extraction (ASE) technique to extract Achyranthes bidentata and to explore the application of this technique in quality control of Chinese materia medica. Methods Investigation of single factor was used to optimize the conditions that affected the efficiency for ASE from A. bidentata by RP-HPLC using ecdysterone as a quantitative marker. Results The optimized conditions for ASE of A. bidentata were obtained as follows: methanol as solvent, particle size between 0.3 and 0.45 mm, temperature at 100 ℃, pressure under 10.34 MPa, 6 min duration and once extraction. Conclusion ASE technique can be used to extract A. bidentata quickly and effectively.
10.Mechanism of DJ -1 against oxidative stress and its roles in the pathogenesis of age -related macular degeneration and Fuchs endothelial corneal dystrophy
Ya-Lan, ZHOU ; Jun, ZENG ; Bai-Hua, CHEN
International Eye Science 2016;16(8):1465-1468
?DJ-1 has been reported to act as aredox-activated chaperone and sensor of oxidative stress participated in a variety of activities in cellular, playing an important role in resisting oxidative stress, regulating signaling pathways and gene transcription, and maintaining mitochondria dynamic balance. DJ -1 is closely related to the occurrence and development of various diseases. Recently, the effect of DJ-1 in eye diseases has drawn more attention, and researchers have found its significant role of resistance to oxidative stress in the pathogenesis of Fuchs endothelial corneal dystrophy ( FECD) and age-related macular degeneration ( AMD ) .This review will state the mechanism of DJ-1 against oxidative stress and its role in the development of eye diseases.