2.Establishment and verification of multiplex fluorescence quantitative PCR method for detection of CHO host cell residual DNA content and fragmentation degree
Chinese Journal of Biologicals 2024;37(12):1495-1504
Objective To establish and verify a multiplex fluorescence quantitative PCR method for detection of CHO host cell residual DNA content and fragmentation degree, so as to provide a reliable method for safety detection of related vaccines.Methods Two sets of primers(Alu-L and Alu) and universal probe were designed for clone250 and clone49c, members of Alu sequence family of CHO cells. The UBI3 gene of capsicum was introduced as external standard gene, and corresponding primers and probe were designed. Multiplex reaction system of Alu sequence and external standard gene was established, and the residual DNA of CHO cells in samples was quantified by the multiplex fluorescence quantitative PCR. The linear range,limit of quantitation(LOQ), specificity, robustness, precision and accuracy of the method were verified. In addition, the DNA fragmentation degree of CHO cells was determined by comparing the △Ct [Ct(Alu-L)-Ct(Alu)] of the amplification curves of long and short primers of Alu sequences. Results The multiplex reaction standard curve had a good linear relationship in the range of 0. 001-0. 1 ng/μL, R~2≥ 0. 99. The addition of external standard gene had no effect on the detection ability for CHO target, and the LOQ was 0. 5 fg/μL. The DNA of human, E.coli, rats and mice had no effect on the detection, the primers targeting residual DNA of CHO cells had no specific amplification in the genomes of seven animal cell lines, and the fragmented DNA samples showed no effect on the detection results. The CVs of precision verification were all less than 15%, and the recoveries of simulated samples in normal saline and PBS containing 1% BSA were in the range of 70%-130%. The△Ct of Alu and Alu-L amplification curves increased with the degree of sample fragmentation, and the fitting curve R~2≥ 0. 99.Conclusion The multiplex fluorescence quantitative PCR detection method established in this study can rapidly and accurately quantify the residual DNA of CHO cells and determine the fragmentation degree of samples according to △Ct, which can be used for the safety detection of the related vaccines.
3.Progress in research on liquichip technology in biomedical
International Journal of Biomedical Engineering 2011;34(6):371-374
Liquichip technology utilizes different fluorescent coding microbead as carriers of bio-probes and flow cytometry as optical detection method,to accomplish bio-molecules reaction in suspension liquid system.This article introduces the application of liquidchip technology in the analysis of autoimmune disease,cytokine,contagious disease,endocrinopathy,neural disease,tumor marker detection and so on.With the advantages of high sensitivity,high speed,high flux,multi-analyte,high accuracy and excellent repeatability,liquichip technology has promising prospect to be widely used as advanced biomedical detection platform.
6.Relationship between CD_(44v) and leukemia
Journal of Leukemia & Lymphoma 2010;19(3):184-187
CD_(44) adhesion molecule is a cell surface transmembrane glycoprotein.It displays many varinat isoforms generated by alternative splicing of exons, including CD_(44s). and CD_(44v). It has been reported that CD_(44) adhesion molecule is strongly expressed on the surface of leukemic blasts in all acute myeloid leukemia subtypes.It has been found through in vitro study that CD_(44) monoclonal antibodies can inhibit proliferation of all acute myeloid leukemia cell lines and induce terminal differentiate or apoptosis of some cell lines to diverse extent. In this review the relationship between CD_(44v), and leukemia was analysed.
7.The relationship of self-management skills and the quality of life among solitary elderly patients with hypertension
Chinese Journal of Practical Nursing 2014;30(2):65-67
Objective To investigate the relationship of self-management skills and the quality of life among solitary elderly patients with hypertension.Methods 110 cases of solitary elderly patients with hypertension were measured with self-designed Self-management of Hypertension Scale and Short Form Health Survey (SF-36) and the correlation of them was analyzed.Results The levels of self-management skills of elderly hypertensive patients was at a middle level,and the mean score of self-management skills was (78.6 ± 5.2) points.The total score of SF-36 of patients with good self-management skills was higher than those with bad self-management skills.The therapy management,catering management,psychosocial management,sports management and overall quality of self-management of elderly patients with hypertension were positively correlated with survival quality.The factors influencing self-management ability of patients incuded education,gender,social resource utilization,psychological conditions.Conclusions Community health nurses should pay attention to self-management skills such as diet,exercise,therapy,social and psychological aspects in elderly hypertensive patients in order to improve their quality of life.Furthermore community nurses should strengthen intervention for solitary eldedy males with lower education level,poor utilization of social resources to improve their self-management skills.
8.Design of Electrocardiosignal Filter
Chinese Medical Equipment Journal 1989;0(04):-
Objective There are many interferences such as power interference, electrode polarization interference, myoelectricity interference, baseline drift, apparatus buzz and so on in electrocardiosignal acquisition system. It is very difficult to effectively remove these interferences in the process of actual measurement. Methods Power harmonic interference, electrode polarization interference and myoelectricity interference were removed by low-pass filter. Baseline drift was removed by high-pass filter. Power basic frequency interference was removed by 50 Hz trap filter. Results These three design proposals had such advantages as good effect, rapid speed and easy realization. Conclusion They can be well applied in ECG monitor with embedded CPU and real time character. Besides, they can also be used in medical apparatus measuring other biological electric signals and industrial observation and control system.
9.Review on Working Principles of Brain Computer Interface System
Chinese Medical Equipment Journal 2004;0(08):-
Actual research on brain-computer interface system is analyzed.The basic structure and operational principle of this system are set forth.Methods of signal collection,feature extraction and selection classification of system signals are concluded.Latest analysis methods and existing defects of feature extraction and selection classification are introduced in detail.Problems existing in brain-computer interface system are also analyzed.Great advantages on feature extraction of wavelet transform & wavelet packet decomposition and wide prospects on selection classification of neural network are pointed out at last.
10.Inhibitory effect and its mechanism of 7-O-succinyl macrolactin A against cell proliferation, invasion and migration in human lung cancer H460 cells
Journal of China Pharmaceutical University 2017;48(1):82-88
This study aimed at investigating the effects and mechanisms of 7-O-succinyl macrolactin A in inhibi-ting human lung cancer. After treatment of human lung cancer cell lines H460 with 7-O-succinyl macrolactin A, MTS assay was employed to determine cell proliferation;crystal violet staining was used to detect cell adhesion of H460;transwell chamber assay and wound healing assay were performed to evaluate cell invasion and migration;and flow cytometry assay was adopted to evaluate cell cycle. Western blotting and real-time PCR were also employed to determine the expression of β-catenin, c-Myc, Cyclin D1, vimentin, N-cadherin, CD44, integrin β1, Bcl-2 , Survivin and MMP-2/9. The phosphorylation of AKT and mTOR was determined as well. In vitro prolifera-tion of H460 was inhibited significantly by 7-O-succinyl macrolactin A. Cell adhesion, invasion and migration abilities were also attenuated. Western blot and real-time PCR showed that the expressions of β-catenin, c-Myc, cyclin D1, vimentin, N-cadherin, CD44, integrin β1, Bcl-2, Survivin and MMP-2/9 were down-regulated by 7-O-succinyl macrolactin A. It was also found that phosphorylation of AKT and mTOR was inhibited by 7-O-succinyl macrolactin A. 7-O-succinyl macrolactin A can inhibit the in vitro growth and invasion of human lung cancer cell lines H460.