1.Analysis on integrity of target nucleic acid in recombinant adeno-associated virus
Chinese Journal of Biologicals 2024;37(6):641-645
Objective To explore a method for detecting the integrity of target nucleic acid in recombinant adeno-associated virus(rAAV),and establish a preliminary analysis algorithm.Methods The free DNA fragment of rAAV was digested,and the virus genome was extracted.Five pairs of overlapping primers were designed,using the orthogonal array method,which were detected by digital PCR respectively,with conventional conditions:20 μL reaction system with EveGreen and4 μL template,and digital PCR conditions:95 ℃ 5 min;95 ℃ 15 s,55 ℃ 30 s,72 ℃ 90 s for 45 cycles.The enhancement condition of ultra-long nucleic acid fragment was 25 μL reaction system with Mix B-2 000 bp and 2 μL template,and the quantitative analysis was performed by using the software attached to the instrument.Using the least square method,the number of full-length fragments was fitted and analyzed,and then the integrity distribution of target nucleic acid was interpreted.Results The fragments with the distance between primers of no more than 1 200 nt were amplified effectively,and a series of effective copies of fragments with a length of about 1 000 nt were obtained by systematic analysis.The copy number of common fragments was fitted and analyzed by the least square method.It was estimated that the full-length fragments in the sample were no more than 1 234 copies/μL,and there was a signficant difference(P < 0.05)between this value and the maximum measured value of 1 443 copies/μL,with the difference of approximately 16.9%.Conclusion A preliminary detection method for the integrity of target nucleic acid in rAAV has been developed,and a certain amount of incomplete target nucleic acids were analyzed in the test sample,laying a foundation for further in-depth research.
2.Research progress and application prospects of circular RNA
Chinese Journal of Biologicals 2025;38(04):473-480+489
Circular RNA(circRNA) is a type of RNA with a circular coding structure, characterized by a circular topological structure formed during the RNA splicing process. Compared to traditional linear mRNA, circRNA exhibits enhanced stability and immunogenicity, leading to its wide-ranging potential applications in areas such as vaccines, biomarkers, and targeted therapies. This paper summarizes the research progress of circRNA, including its synthesis and degradation in vivo and in vitro, components, biological functions, and applications, aiming to provide novel insights for their utilization in vaccine design and development.
3.Detection and Significance of Class Ⅱ Integrons in Clinically Isolated Strains of Stenotrophomonas maltophilia
Xiaowei DENG ; Changting LIU ; Tianzhi LI ; Junfeng WANG ; Liwu LIANG ; Yanping HUANG ; Chen MO
Chinese Journal of Nosocomiology 2006;0(09):-
OBJECTIVE To investigate the distribution and characterization of the classes Ⅰ,Ⅱ and Ⅲ integrons on Stenotrophomonas maltophilia and clarify their influence on the bacterial drug-resistance.METHODS A multi-PCR assay using specific primers of int1,int2 and int3 was constructed to screen classes Ⅰ,Ⅱ and Ⅲ integrons.RESULTS Class Ⅰ integron was detected in 13.4% of clinical isolates,3 isolates harbored among class Ⅱ integrons. There was not been reported in abroad.CONCLUSIONS Classes Ⅰ and Ⅱ integrons could play an important role in causing the antibiotic multidrug resistance.
4.Noncoding RNAs in cancer and cancer stem cells.
Tianzhi HUANG ; Angel ALVAREZ ; Bo HU ; Shi-Yuan CHENG
Chinese Journal of Cancer 2013;32(11):582-593
In recent years, it has become increasingly apparent that noncoding RNAs (ncRNA) are of crucial importance for human cancer. The functional relevance of ncRNAs is particularly evident for microRNAs (miRNAs) and long noncoding RNAs (lncRNAs). miRNAs are endogenously expressed small RNA sequences that act as post-transcriptional regulators of gene expression and have been extensively studied for their roles in cancers, whereas lncRNAs are emerging as important players in the cancer paradigm in recent years. These noncoding genes are often aberrantly expressed in a variety of human cancers. However, the biological functions of most ncRNAs remain largely unknown. Recently, evidence has begun to accumulate describing how ncRNAs are dysregulated in cancer and cancer stem cells, a subset of cancer cells harboring self-renewal and differentiation capacities. These studies provide insight into the functional roles that ncRNAs play in tumor initiation, progression, and resistance to therapies, and they suggest ncRNAs as attractive therapeutic targets and potentially useful diagnostic tools.
Gene Expression Regulation, Neoplastic
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Humans
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MicroRNAs
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genetics
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metabolism
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Neoplasms
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genetics
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metabolism
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pathology
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therapy
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Neoplastic Stem Cells
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metabolism
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RNA, Long Noncoding
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genetics
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metabolism
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RNA, Untranslated
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genetics
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metabolism
5.A survey on clinical applications of transradial access for peripheral interventions in 2022
Xin ZHOU ; Minjie YANG ; Xiaoqian MENG ; Jiarui LI ; Yonghui HUANG ; Tianzhi AN ; Jian YOU ; Ersheng LI ; Wen ZHANG ; Zhiping YAN
Chinese Journal of Clinical Medicine 2024;31(3):440-444
Objective To investigate the current status,obstacles,and specific needs associated with the application of transradial access(TRA)in peripheral interventions in Chinese hospitals,with the aim of promoting the broader adoption of TRA in interventional procedures.Methods The Committee of Interventional Oncology of China Anti-Cancer Association conducted a cross-sectional questionnaire survey to investigate and analyze the nationwide situation of TRA in peripheral interventional surgeries in 2022.Results Personnel from 60 hospitals participated in the questionnaire.The results showed that the number of peripheral interventionals performed using TRA was significantly lower than that performed using transfemoral access(TFA),with considerable variability among hospitals.The primary obstacles to the adoption of TRA were the lack of suitable catheters and the difficulty of radial artery puncture.Approximately 86.67%of the institutions expressed a high expectation for the innovation of new TRA-specific devices,particularly catheters,microcatheters,and guidewires.60.00%of medical institutions indicated a strong desire for systematic training and participation in multi-center clinical trials.Conclusions TRA peripheral intervention is feasible in many hospitals in China,however,systematic training and further promotion of TRA are essential.The innovation of new TRA-specific devices and TRA in peripheral intervention is urgent.