1.Application and Biological Evaluation of Magnetic Cell Sorting Technology
Tian HONG ; Jing-Wen LI ; Ren-Ai LI ; Er-Ning CHEN ; Lu-Lu ZHAO ; Mei-Hong DU
Progress in Biochemistry and Biophysics 2024;51(1):123-135
Magnetic cell sorting technology is a highly specific and rapid cell sorting technology using superparamagnetic nanocomposites for cell sorting, which is widely used in immunology, stem cytology, oncology, clinical medicine and other fields. Magnetic cell sorting technology is divided into positive isolation, negative isolation/untouched cell isolation, depletion, multi-step isolation and automated cell separation systems. In this review, we firstly give a brief introduction to the classification and application of magnetic cell sorting technology, then discuss several new techniques and challenges based on magnetic cell sorting in recent years, such as improving the sorting efficiency by improving the structure of magnetic materials and magnetic field structure. The necessity of biological evaluation of magnetic cell sorting products was emphatically analyzed. Through the biological evaluation, the advantages and disadvantages of magnetic cell sorting products can be understood, and the research and development ability could be improved. Therefore, 10 biological evaluation technical parameters related to magnetic cell sorting products were proposed: yield, purity, sterility, cytotoxicity, cell morphology, viability, light scattering characteristics of cells, fluorescent antibody labeling ability of cells, cell activation and cell proliferation. The 10 biological evaluation technical parameters play an important role in promoting the standardized application of magnetic cell sorting.
2.Principle and Application of Ultraviolet Crosslinking Immunoprecipitation Technology
Ya-Qiong DU ; Wan-Yao WANG ; Fan GAO ; Yang XU ; Wen-Tao SHI
Progress in Biochemistry and Biophysics 2024;51(1):136-144
The UV cross-linking immunoprecipitation (CLIP) technique was first established in 2003. Sequences of target RNAs and binding sites of specific RNA-binding proteins (RBPs) were identified within the entire transcriptome by UV cross-linking, immunoprecipitation, reverse transcription, and subsequent high-throughput sequencing. Over the last 20 years, CLIP has been continuously modified and improved. Advanced operability and accuracy have extended its application category. Currently, the widely used CLIP technologies include high-throughput sequencing with crosslinking-immunoprecipitation (HITS-CLIP), photoactivatable-ribonucleoside-enhanced CLIP (PAR-CLIP), individual nucleotide resolution CLIP (iCLIP), enhanced CLIP (eCLIP), infrared-CLIP (irCLIP), etc. HITS-CLIP combines high-throughput sequencing with UV cross-linking immunoprecipitation. The 254 nm UV cross-linking and RNAase digestion steps allow the technology to capture transient intracellular RBP-RNA interactions. However, there are limitations in the efficiency of UV cross-linking, with low resolution and high intrinsic background noise. For PAR-CLIP, photoactivatable ribonucleoside was incorporated into RNA molecules, and RBP cross-linked with RNA by 365 nm UV light to improve cross-linking efficiency and resolution. Cross-linking mediated single-base mutations provide more accurate binding site information and reduce interference from background sequences. Long-term alternative nucleotide incorporation, on the other hand, can be cytotoxic and may skew experimental results. iCLIP can identify RBP-RNA cross-linking sites at the single nucleotide level through cDNA circularization and subsequent re-linearization steps, but it has more experimental procedures, and partial cDNAs lost in the circularization step are inevitable. eCLIP discards the radioisotope labeling procedure and reduces RNA loss by ligating adaptors in two separate steps, greatly improving the library-building efficiency, and reducing bias associated with PCR amplification; however, the efficiency of immunoprecipitation cannot be visually assessed at the early stage of the experiment. The irCLIP technique replaces radioisotopes with infrared dyes and greatly reduces the initial number of cells required for the experiment; however, an infrared imaging scanner is essential for the irCLIP application. To address more particular scientific issues, derivative CLIP-related techniques such as PAPERCLIP, cTag-PAPERCLIP, hiCLIP, and tiCLIP have also been developed in recent years. In practice, the aforementioned CLIP approaches have their advantages and disadvantages. When deciding on a technical strategy, we should take into account our experimental objectives and conditions, such as whether we need to precisely define the RNA site for binding to RBP; whether we have the necessary experimental conditions for working with radioisotopes or performing infrared imaging; the amount of initial sample size, and so on. In addition, the CLIP technique has a relatively large number of procedures and can be divided into several successive experimental modules. We can try to combine modules from different mainstream CLIP technologies to meet our experimental requirements, which also gives us more opportunities to improve and refine them and to build more targeted derivative CLIP technologies according to our research objectives.
3.Research progress of relationship in long-novel coronavirus syndrome and sarcopenia
Wen ZHONG ; Hua'nan JIA ; Chuanfeng LIU ; Wei HUANG ; Jian DU ; Mengjun CHEN
Chinese Critical Care Medicine 2024;36(1):106-109
At present, the major public health challenges caused by novel coronavirus infection have gradually subside. However, a large number of people are still suffering from long-novel coronavirus syndrome or post-novel coronavirus syndrome. The clinical manifestations of long coronavirus syndrome are related to multiple systems, such as respiratory, circulatory, nervous, digestive and musculoskeletal systems, with various long-term persistent symptoms after novel coronavirus infection. At the same time, the infection of the novel coronavirus is an important cause of frailty and sarcopenia in the elderly population. However, at present, the scholars have not paid enough attention to the skeletal muscle weakness caused by the novel coronavirus. Therefore, this paper focuses on the long-novel coronavirus syndrome and sarcopenia to explore the pathological mechanism of skeletal muscle attenuation caused by the SARS-CoV-2 mediated "cytokine storm", mitochondrial damage, hypoxia state and other links, so as to raise the attention of clinical and academic researchers and improve the clinical strategy of frailty and sarcopenia after novel coronavirus infection.
4.Predicting the Risk of Arterial Stiffness in Coal Miners Based on Different Machine Learning Models.
Qian Wei CHEN ; Xue Zan HUANG ; Yu DING ; Feng Ren ZHU ; Jia WANG ; Yuan Jie ZOU ; Yuan Zhen DU ; Ya Jun ZHANG ; Zi Wen HUI ; Feng Lin ZHU ; Min MU
Biomedical and Environmental Sciences 2024;37(1):108-111
5.Relationship between the expression of biomarkers in the activated innate immune cell and vital organs injuries during the cardiopulmonary bypass
Wen XIONG ; Jinghan ZHANG ; Lei DU ; Xiaoling HU
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2024;31(11):1679-1683
Although great progress has been achieved in the techniques and materials of cardiopulmonary bypass (CPB), cardiac surgery under CPB is still one of the surgeries with the highest complication rate. The systemic inflammatory response is an important cause of complications, mainly characterized by activation of innate immune cells and platelets, and up-regulation of inflammatory cytokines. After activation, a variety of molecules on the membrane surface are up-regulated or down-regulated, which can amplify tissue inflammatory damage by releasing cytoplasmic protease and reactive oxygen species, and activate multiple inflammatory signaling pathways in the cell, ultimately leading to organ dysfunction. Therefore, the expression of these cell membrane activation markers is not only a marker of cell activation, but also plays an important role in the process of vital organ injury after surgery. Identification of these specific activation markers is of great significance to elucidate the mechanisms related to organ injury and to find new prevention and treatment methods. This article will review the relationship between these activated biomarkers in the innate immune cells and vital organ injuries under CPB.
6.Preliminary study of the transport and absorption mechanism of Astragalus polysaccharide-Ⅱ on M cells
Wan-wan LÜ ; Ke LI ; Shi-hong FENG ; Yu-wei WEN ; Xue-mei QIN ; Yu-guang DU ; Zhen-yu LI
Acta Pharmaceutica Sinica 2024;59(10):2820-2827
To explore the absorption mechanism of APS-Ⅱ
7.Research advances of pentacyclic triterpenoid natural products
Wen-xin DING ; Bai-lin DU ; Jiao LI ; Chun-lin ZHUANG
Acta Pharmaceutica Sinica 2024;59(5):1163-1175
Pentacyclic triterpenoids are a class of widespread natural compounds containing six isoprene structures with a wide range of pharmacological activities, including antibacterial, anti-inflammatory, antiviral, antitumor, immune regulation,
8.Three 2,3-diketoquinoxaline alkaloids with hepatoprotective activity from Heterosmilax yunnanensis
Rong-rong DU ; Xin-yi GUO ; Wen-jie QIN ; Hua SUN ; Xiu-mei DUAN ; Xiang YUAN ; Ya-nan YANG ; Kun LI ; Pei-cheng ZHANG
Acta Pharmaceutica Sinica 2024;59(2):413-417
Three 2,3-diketoquinoxaline alkaloids were isolated from
9.Vaccination certificate verification for children enrolled in kindergarten and primary school in Guizhou Province from 2020 to 2022
QIAO Sha, TANG Ning, DU Wen, DAI Lifang, DING Ling
Chinese Journal of School Health 2024;45(4):589-592
Objective:
To evaluate the implementation of vaccination certificate verification in Guizhou Province from 2020 to 2022, so as to provide reference for improving the efficiency of vaccination certificate verification and vaccine re inoculation work.
Methods:
Data was drawn from the 2020-2022 report on the verification of vaccination certificates for children entering daycare and enrollment in various cities and prefectures in Guizhou Province. In July, 2021, Guizhou Province began to implement a new inspection scheme with close cooperation between health and education departments, moving forward the gateway, parents using "Guizhou CDC" WeChat official account for self inspection, and a long term supervision and assessment mechanism. A comparative analysis was conducted on the evaluation of vaccination certificate verification rate, vaccination certificate holding rate, full revaccination rate of the National Immunization Program (NIP) for children and full vaccination rate of the NIP vaccine before(2020) and after(2021 and 2022) the implementation of the new plan. Chi square test was used for statistical analysis.
Results:
The rate of vaccination certificate verification of children enrolled in kindergarten and primary school in Guizhou Province increased from 99.85% in 2020 to 100% in 2022, the rate of holding certificate increased from 99.55% in 2020 to 99.91% in 2022, the rate of full vaccination NIP vaccines for kindergarten and primary school entry increased from 78.95% in 2020 to 96.59% in 2022, and the rate of full revaccination increased from 42.40% in 2020 to 79.19% in 2022 ( χ 2=2 203.19, 3 651.67, 291 896.31, 103 938.76, P < 0.01 ).
Conclusions
From 2020 to 2022, the rates of full vaccination and the full revaccination for NIP vaccine among children entering kindergarten in Guizhou Province have increased year by year. Each region should fully utilize the achievements of immunization planning informatization construction to establish effective inspection work ideas, and ensure that eligible children complete the full vaccination process of the national immunization plan vaccine.
10.Pathological Characteristics and Classification of Unstable Coronary Atheroscle-rotic Plaques
Yun-Hong XING ; Yang LI ; Wen-Zheng WANG ; Liang-Liang WANG ; Le-Le SUN ; Qiu-Xiang DU ; Jie CAO ; Guang-Long HE ; Jun-Hong SUN
Journal of Forensic Medicine 2024;40(1):59-63
Important forensic diagnostic indicators of sudden death in coronary atherosclerotic heart dis-ease,such as acute or chronic myocardial ischemic changes,sometimes make it difficult to locate the ischemic site due to the short death process,the lack of tissue reaction time.In some cases,the de-ceased died of sudden death on the first-episode,resulting in difficulty for medical examiners to make an accurate diagnosis.However,clinical studies on coronary instability plaque revealed the key role of coronary spasm and thrombosis caused by their lesions in sudden coronary death process.This paper mainly summarizes the pathological characteristics of unstable coronary plaque based on clinical medi-cal research,including plaque rupture,plaque erosion and calcified nodules,as well as the influencing factors leading to plaque instability,and briefly describes the research progress and technique of the atherosclerotic plaques,in order to improve the study on the mechanism of sudden coronary death and improve the accuracy of the forensic diagnosis of sudden coronary death by diagnosing different patho-logic states of coronary atherosclerotic plaques.


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