1.Establishment of real-time fluorescence quantitative PCR method for detecting the N subgenome of SARS-CoV-2
Taoli HAN ; Zhi ZHANG ; Jiaxin ZHAO ; Pan LU ; Yang JIAO ; Jianhong ZHAO ; Yan GAO ; Shiyao ZHANG ; Kuankuan LIU ; Yujie LIU ; Ru FAN ; Wenjing LI ; Lingli SUN
Chinese Journal of Experimental and Clinical Virology 2025;39(1):96-101
Objective:To establish a fluorescent quantitative RT-PCR assay based on N_sgRNA of SARS-CoV-2 and preliminarily apply it on real samples.Methods:Recombinant plasmid, specific primers and probes of N_sgRNA were designed and synthesized based on Wuhan-Hu-1/2019_MN908947 and synthesis mechanism of subgenomic RNA (sgRNA). Using recombinant plasmid as amplification templates, the optimal reaction conditions and reaction system were screened according to the Ct value, fluorescence intensity, and shape of amplification curve and was evaluated for sensitivity, reproducibility, and specificity. Meanwhile, the possibility of practical application of the method was explored by testing 172 clinical samples and 256 municipal wastewater samples. Results:A qRT-PCR assay for N_sgRNA in SARS-CoV-2 was initially established. The detection limit of the assay was 20 copies/mL, and the variation coefficients of in-batch (0.002%~0.767%) and batch to batch repetition (0.016%~0.752%) were less than 1%. Only N_sgRNA recombinant plasmid was detected in the specificity assay. So the method is more highly sensitive, specific and reproducible. The RatiosgRNA/ gRNA of clinical samples HK.3, EG.5.1, JN.1 and their sub-lineages and their corresponding urban sewage samples in epidemic period were significantly different ( P<0.05). There is a strong correlation between the median of RatiosgRNA/ gRNA in clinical samples and sewage samples in the same period (correlation coefficient r=1.000, P=0.010). Conclusions:In this study, a qRT-PCR method for detecting N_sgRNA of SARS-CoV-2 was established and the method has the characteristics of higher sensitivity, stronger specificity and better repeatability, and it can be used to detect SARS-CoV-2 infectivity.
2.Establishment of a rapid detection method for SARS-CoV-2 Omicron variant nucleic acid based on BMD-PCR technology
Yang JIAO ; Taoli HAN ; Jianhong ZHAO ; Shiyao ZHANG ; Zhi LYU ; Shaocheng WANG ; Lingli SUN
Chinese Journal of Experimental and Clinical Virology 2025;39(1):109-114
Objective:The SARS-CoV-2 has a high natural mutation rate, and dynamic monitoring of virus variants remains a key focus in current COVID-19 prevention and control efforts.Methods:In this study, a sensitive and rapid method for detecting SARS-CoV-2 omicron variant nucleic acid was established based on the BMD-PCR technology.Results:This method showed good specificity, and had no cross-reactivity with 11 common viruses transmitted via the respiratory and gastrointestinal tracts, and the limit of detection is 555 copies/ml. Compared with SARS-CoV-2 whole-genome sequencing result, among 50 samples with original Ct values ≤32 tested for the Omicron variant, 49 samples tested positive for the N679K mutation site using BMD-PCR Omicron variant detection, achieving a concordance rate of 98.00%. For 30 samples JN.1 lineage, 29 samples tested positive for the K356T mutation site using BMD-PCR JN.1 lineage detection, with a concordance rate of 96.67%. For 10 samples with original SARS-CoV-2 detection Ct values between 35 and 32, 7 samples tested positive for the N679K mutation site using BMD-PCR Omicron variant detection, Resultsing in a detection rate of 70.00%. For samples with SARS-CoV-2 nucleic acid detection Ct values>35, the detection rate for the N679K mutation site in the BMD-PCR Omicron variant was 20.00%.Conclusions:This method can serve as a high-throughput supplementary approach for the preliminary identification of SARS-CoV-2 variant genotypes.
3.Visualization Analysis on Research Literature about Astragalus Polysaccharides from 2013 to 2023
Hong LI ; Liu LI ; Qiuqing HUANG ; Shiyao YANG ; Junju ZOU ; Fan XIAO ; Qin XIANG ; Xiu LIU ; Yanling FU ; Yongjun WU ; Rong YU
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(6):73-79
Objective To analyze the research status and hotspots in the field of astragalus polysaccharides;To provide references for further research.Methods Research literature about astragalus polysaccharides was retrieved from CNKI,Wanfang Data,VIP,PubMed,and Web of Science databases from 1st,Jan.2013 to 1st,July 2023.NoteExpress 3.7 software was used to manage the literature and ultimately establish a database.Excel 2019,CiteSpace 6.2.2R and VOSviewer 1.6.18 were used to visually analyze the publication volume,authors,institutions,and keywords of the included literature.Results A total of 2 462 articles were included,with 1 284 Chinese articles and 1 178 English articles.The main research institutions were Gansu University of Chinese Medicine,Shandong University of Traditional Chinese Medicine,and Beijing University of Chinese Medicine.The core authors of Chinese literature were Liu Yongqi,Wang Hongxin,Lu Meili,etc.The core authors of English literature included Zhang Wei,Li Ke,Yang Xiaojun,etc.High-frequency keywords of Chinese literature included Astragali Radix,rats,polysaccharides,cell apoptosis,and oxidative stress,etc.High frequency keywords in English literature included expression,in vitro,oxidative stress,apoptosis,etc.Conclusion The research on astragalus polysaccharides focuses on their pharmacological effects and mechanisms.Intestinal flora,immune regulation,autophagy and apoptosis are the hot action mechanisms in this field.The focus of disease research involves tumor and diabetes,and antiviral,anti infection and other pharmacological effects are the research trend.
4.Analysis and discussion of the common problems found in medical device clinical trials inspection results from 2016 to 2023
Yuyan LIANG ; Fang JI ; Ying PAN ; Shiyao XU ; Shu YANG ; Liang XU
Chinese Journal of Clinical Pharmacology and Therapeutics 2025;30(1):131-137
By collecting inspection results from medical device clinical trials from 2016 to 2023,to analyze and discuss common problems of clinical trials from both in vitro diagnostic reagents and medical devices,and propose the suggested mea-sures taken by participants in the clinical trial,so as to avoid similar problems occur and ensure the quality of clinical trials.
5.Chain mediation of oncology nurses' general self-efficacy and humanistic care competence in empathy and medical narrative competence
Yong DONG ; Shuaizhong CAI ; Ruie LI ; Wentong ZHOU ; Lei JIANG ; Haiyin YANG ; Shiyao YANG ; Yuexi ZHU ; Jinhu MIAO ; Qiongyao GUAN
Chinese Journal of Modern Nursing 2025;31(8):1072-1077
Objective:To explore the chain mediating role of general self-efficacy and humanistic care competence between oncology nurses' empathy and medical narrative competence, with a view to informing the development of targeted medical narrative competence enhancement strategies.Methods:Convenience sampling was used to select 1 104 oncology nurses from 17 ClassⅢ Grade A hospitals in Yunnan Province from May to August 2023 for the study. The General Information Questionnaire, Narrative Competence Scale, Chinese version of the Jefferson Scale of Empathy, Caring Ability Inventory, and General Self-Efficacy Scale were used to survey oncology nurses. Pearson correlation was used to analyze the correlation among oncology nurses' general self-efficacy, empathy, humanistic care competence, and medical narrative competence. Mediating effects were analyzed using Model 6 in the SPSS macro program PROCESS 4.0 plug-in. Bootstrap method was used to repeat the sampling 5 000 times for the chained mediating effect test.Results:A total of 1 104 questionnaires were distributed and the number of valid questionnaires recovered was 931, with a valid recovery rate of 84.33% (931/1 104). Among 931 oncology nurses, the scores of Narrative Competence Scale, Caring Ability Inventory, Chinese version of Jefferson Scale of Empathy, and General Self-Efficacy Scale were (151.52±17.61), (191.68±18.80), (95.31±13.96), and (27.59±6.02), respectively. There was a positive two-by-two correlation among oncology nurses' medical narrative competence, empathy, humanistic care competence, and general self-efficacy ( P<0.01). General self-efficacy and humanistic care competence acted as chain mediators between oncology nurses' empathy and medical narrative competence, with the mediating effect accounting for 43.13% of the total effect. Conclusions:General self-efficacy and humanistic care are mediating variables between oncology nurses' empathy and medical narrative competence. Nursing managers should focus on the cultivation of nurses' empathy, stimulate nurses' self-efficacy, and promote the practice of nursing humanistic care, which in turn improves oncology nurses' medical narrative competence and the quality of nursing services.
6.Visualization Analysis on Research Literature about Astragalus Polysaccharides from 2013 to 2023
Hong LI ; Liu LI ; Qiuqing HUANG ; Shiyao YANG ; Junju ZOU ; Fan XIAO ; Qin XIANG ; Xiu LIU ; Yanling FU ; Yongjun WU ; Rong YU
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(6):73-79
Objective To analyze the research status and hotspots in the field of astragalus polysaccharides;To provide references for further research.Methods Research literature about astragalus polysaccharides was retrieved from CNKI,Wanfang Data,VIP,PubMed,and Web of Science databases from 1st,Jan.2013 to 1st,July 2023.NoteExpress 3.7 software was used to manage the literature and ultimately establish a database.Excel 2019,CiteSpace 6.2.2R and VOSviewer 1.6.18 were used to visually analyze the publication volume,authors,institutions,and keywords of the included literature.Results A total of 2 462 articles were included,with 1 284 Chinese articles and 1 178 English articles.The main research institutions were Gansu University of Chinese Medicine,Shandong University of Traditional Chinese Medicine,and Beijing University of Chinese Medicine.The core authors of Chinese literature were Liu Yongqi,Wang Hongxin,Lu Meili,etc.The core authors of English literature included Zhang Wei,Li Ke,Yang Xiaojun,etc.High-frequency keywords of Chinese literature included Astragali Radix,rats,polysaccharides,cell apoptosis,and oxidative stress,etc.High frequency keywords in English literature included expression,in vitro,oxidative stress,apoptosis,etc.Conclusion The research on astragalus polysaccharides focuses on their pharmacological effects and mechanisms.Intestinal flora,immune regulation,autophagy and apoptosis are the hot action mechanisms in this field.The focus of disease research involves tumor and diabetes,and antiviral,anti infection and other pharmacological effects are the research trend.
7.Establishment of real-time fluorescence quantitative PCR method for detecting the N subgenome of SARS-CoV-2
Taoli HAN ; Zhi ZHANG ; Jiaxin ZHAO ; Pan LU ; Yang JIAO ; Jianhong ZHAO ; Yan GAO ; Shiyao ZHANG ; Kuankuan LIU ; Yujie LIU ; Ru FAN ; Wenjing LI ; Lingli SUN
Chinese Journal of Experimental and Clinical Virology 2025;39(1):96-101
Objective:To establish a fluorescent quantitative RT-PCR assay based on N_sgRNA of SARS-CoV-2 and preliminarily apply it on real samples.Methods:Recombinant plasmid, specific primers and probes of N_sgRNA were designed and synthesized based on Wuhan-Hu-1/2019_MN908947 and synthesis mechanism of subgenomic RNA (sgRNA). Using recombinant plasmid as amplification templates, the optimal reaction conditions and reaction system were screened according to the Ct value, fluorescence intensity, and shape of amplification curve and was evaluated for sensitivity, reproducibility, and specificity. Meanwhile, the possibility of practical application of the method was explored by testing 172 clinical samples and 256 municipal wastewater samples. Results:A qRT-PCR assay for N_sgRNA in SARS-CoV-2 was initially established. The detection limit of the assay was 20 copies/mL, and the variation coefficients of in-batch (0.002%~0.767%) and batch to batch repetition (0.016%~0.752%) were less than 1%. Only N_sgRNA recombinant plasmid was detected in the specificity assay. So the method is more highly sensitive, specific and reproducible. The RatiosgRNA/ gRNA of clinical samples HK.3, EG.5.1, JN.1 and their sub-lineages and their corresponding urban sewage samples in epidemic period were significantly different ( P<0.05). There is a strong correlation between the median of RatiosgRNA/ gRNA in clinical samples and sewage samples in the same period (correlation coefficient r=1.000, P=0.010). Conclusions:In this study, a qRT-PCR method for detecting N_sgRNA of SARS-CoV-2 was established and the method has the characteristics of higher sensitivity, stronger specificity and better repeatability, and it can be used to detect SARS-CoV-2 infectivity.
8.Establishment of a rapid detection method for SARS-CoV-2 Omicron variant nucleic acid based on BMD-PCR technology
Yang JIAO ; Taoli HAN ; Jianhong ZHAO ; Shiyao ZHANG ; Zhi LYU ; Shaocheng WANG ; Lingli SUN
Chinese Journal of Experimental and Clinical Virology 2025;39(1):109-114
Objective:The SARS-CoV-2 has a high natural mutation rate, and dynamic monitoring of virus variants remains a key focus in current COVID-19 prevention and control efforts.Methods:In this study, a sensitive and rapid method for detecting SARS-CoV-2 omicron variant nucleic acid was established based on the BMD-PCR technology.Results:This method showed good specificity, and had no cross-reactivity with 11 common viruses transmitted via the respiratory and gastrointestinal tracts, and the limit of detection is 555 copies/ml. Compared with SARS-CoV-2 whole-genome sequencing result, among 50 samples with original Ct values ≤32 tested for the Omicron variant, 49 samples tested positive for the N679K mutation site using BMD-PCR Omicron variant detection, achieving a concordance rate of 98.00%. For 30 samples JN.1 lineage, 29 samples tested positive for the K356T mutation site using BMD-PCR JN.1 lineage detection, with a concordance rate of 96.67%. For 10 samples with original SARS-CoV-2 detection Ct values between 35 and 32, 7 samples tested positive for the N679K mutation site using BMD-PCR Omicron variant detection, Resultsing in a detection rate of 70.00%. For samples with SARS-CoV-2 nucleic acid detection Ct values>35, the detection rate for the N679K mutation site in the BMD-PCR Omicron variant was 20.00%.Conclusions:This method can serve as a high-throughput supplementary approach for the preliminary identification of SARS-CoV-2 variant genotypes.
9.Mediating Role of Psychological Resilience between Perceived Spousal Support and Dignity Loss in Breast Cancer Patients
Haiyin YANG ; Yong DONG ; Shiyao YANG ; Yuexi ZHU ; Yuhan SHEN ; Jinhu MIAO ; Qiongyao GUAN
Journal of Kunming Medical University 2025;46(3):164-170
Objective To explore the mediating effect of psychological resilience between perceived spousal support and dignity loss in breast cancer patients.Methods A convenience sampling method was employed,involving 377 breast cancer patients who met the study criteria and received treatment at a tertiary oncology hospital in Yunnan Province from March to September 2023.Data were collected using a general information questionnaire,the Dignity Scale,the Perceived Spousal Support Scale,and the Psychological Resilience Scale.Data analysis was conducted using IBM SPSS 26.0 software.Results The scores for dignity loss,perceived spousal support,and psychological resilience of breast cancer patients were(41.72±6.77),(100.42±6.93),and(75.27±8.50),respectively.Pearson correlation analysis showed that dignity loss was negatively correlated with both perceived spousal support and psychological resilience(r1=-0.568,r2=-0.640,both P<0.05).Mediation effect analysis indicated that psychological resilience had a mediating effect between perceived spousal support and dignity loss,with a value of-0.320(95%CI-0.409 to-0.246),accounting for 57.1%of the total effect.Conclusion The dignity of breast cancer patients in this study is in a state of mild loss,and psychological resilience plays a partial mediating role between perceived spousal support and dignity loss.This suggests that clinical medical staff should pay attention to the spousal support situation of breast cancer patients,provide timely guidance and health education to spouses,enhance the level of support from spouses,and strengthen the confidence and capability of couples in jointly coping with the disease,thereby alleviating dignity loss in breast cancer patients and improving their quality of life.
10.A Systematic Evaluation of Incidence and Influencing Factors of Falls in Adult Cancer Patients
Shiyao YANG ; Yanxia YANG ; Haiyin YANG ; Yong DONG ; Jinhu MIAO ; Yuexi ZHU ; Qiongyao GUAN
Journal of Kunming Medical University 2025;46(5):101-109
Objective To systematically evaluate the incidence and influencing factors of falls in adult cancer patients,and provide evidence support for early identification and prevention of falls.Methods Eight Chinese and English databases,including PubMed,Embase,CNKI,and Wanfang Database,were searched by computer.The databases were established until January 2024 and included in cross-sectional,cohort,and case-control studies based on the incidence and/or influencing factors of falls in adult cancer patients.Two researchers independently screened and reviewed literature,evaluated literature quality,and extracted data before conducting meta-analysis using RevMan 5.4 software.Results A total of 19 articles were incorporated into the research,with a total sample population size of 70508 cases.The results showed that the incidence of comorbid falls in adult tumor patients was 23.0%(95%CI 0.23,0.24).Age,female,low education level,fall history,fall fear,middle and late stage of tumor,tumor type(breast cancer,reproductive system tumor,prostate cancer),comorbidity,chemotherapy,radiotherapy,multiple drugs(n≥5),drugs(antidepressants,antipsychotics,sedatives and hypnotics),related symptoms(fever,peripheral neuropathy,fatigue,depression),malnutrition,low weight,cognitive dysfunction,balance disorders,gait abnormalities,and low ability of daily living were the influencing factors for falls of adult tumor patients.Conclusion Cancer professionals should identify the influencing factors of falls early and take targeted measures to reduce the occurrence of falls.

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