1.Isolation and characterization of an epidemic ON1 genotype of respiratory syncytial virus subtype A in Beijing
Xuchang SHAN ; Jiao REN ; Wei ZHANG ; Zhongxian ZHANG ; Lingfang ZHANG ; Jia LI ; Qiaohong CHU ; Jingdong SONG ; Zhihai CHEN ; Yao DENG ; Desheng ZHAI ; Wenjie TAN
Chinese Journal of Microbiology and Immunology 2025;45(1):10-16
Objective:To obtain a prevalent respiratory syncytial virus (RSV) clinical isolate in Beijing and analyze the genotype and biological characteristics of the strain.Methods:A nasopharyngeal secretion specimen was collected from a child with RSV infection in Beijing in 2023 and used for viral isolation. Viral nucleic acid was amplified using qRT-PCR. The isolated virus was identified by transmission electron microscopy, indirect immunofluorescence assay, and plaque formation assay. A phylogenetic analysis was conducted based on the whole-genome sequencing results. Virus titers were determined, and replication characteristics were analyzed. The efficacy of the isolated strain for in vitro screening of antiviral drugs was validated. Results:A clinical RSV isolate, named hRSV/C-Tan/BJ 202301, was successfully isolated, which could form syncytia in Hep-2 cells. Spherical, filamentous, and irregular virus particles were observed by electron microscopy. Immunofluorescence detection showed green fluorescence in Hep-2 cells, and plaque assay showed round plaques, which were similar to the Long strain in morphology. Genomic sequence analysis showed that it belonged to ON1 genotype. It exhibited similar cell growth kinetics characteristics with the Long strain and could be used for antiviral drug screening in vitro. Conclusions:In this study, one RSV strain is successfully isolated and identified. The biological characteristics and the phylogenetic relationship of this strain reflect the characteristics of the circulating strains in Beijing, which provides experimental material for RSV vaccine development and antiviral drug screening in China.
2.Development and application of an optimized focus-forming assay for quantitation of influenza A virus titer
Jia LI ; Qiaohong CHU ; Lingfang ZHANG ; Xuchang SHAN ; Tangqi WANG ; Ruiwen HAN ; Yujie JIANG ; Donghong WANG ; Baoying HUANG ; Yao DENG ; Wenjie TAN
Chinese Journal of Microbiology and Immunology 2025;45(1):45-52
Objective:To establish and optimize a novel method, focus forming assay (FFA), for quantitation of influenza A virus (FluA) and compare its application performance with traditional plague forming assay (PFA).Methods:The foci chromogenic effects of three peroxidase substrates in immunostaining were compared. The PFA and FFA methods were used to explore FluA incubation times and plaque morphology on 12-well plates, and to determine optimal incubation times and virus adsorption volumes for different FluA subtypes on 96-well plates. The correlation between FFA and PFA was evaluated, and the optimized FFA was applied to the in vitro antiviral efficacy analysis of Favipiravir and neutralization test against different subtypes of FluA. Results:TRUEBLUE substrate was identified as the optimal substrate for foci visualization. Compared with the PFA, the FFA showed improved sensitivity and reduced detection time in FluA titration, and good correlation was shown between the two methods′ results. By replacing the 96-well plate with the 12-well plate for FFA titration of different subtypes of FluA, the detection time was shortened, and the amount of serum samples used could be further reduced by optimizing the virus adsorption volume. The half-maximal effective concentration of favipiravir against influenza viruses assessed by the FFA and PFA methods showed no significant difference, and was consistent with the results obtained from quantitative PCR. Additionally, the focus reduction neutralization test and hemagglutination inhibition assays demonstrated strong correlation in determining antibody titers against FluA in serum neutralization assays.Conclusions:The improved FFA method developed here provides a more efficient experimental tool for FluA titration, antiviral drug screening and broad-spectrum vaccine evaluation.
3.Isolation and characterization of an epidemic ON1 genotype of respiratory syncytial virus subtype A in Beijing
Xuchang SHAN ; Jiao REN ; Wei ZHANG ; Zhongxian ZHANG ; Lingfang ZHANG ; Jia LI ; Qiaohong CHU ; Jingdong SONG ; Zhihai CHEN ; Yao DENG ; Desheng ZHAI ; Wenjie TAN
Chinese Journal of Microbiology and Immunology 2025;45(1):10-16
Objective:To obtain a prevalent respiratory syncytial virus (RSV) clinical isolate in Beijing and analyze the genotype and biological characteristics of the strain.Methods:A nasopharyngeal secretion specimen was collected from a child with RSV infection in Beijing in 2023 and used for viral isolation. Viral nucleic acid was amplified using qRT-PCR. The isolated virus was identified by transmission electron microscopy, indirect immunofluorescence assay, and plaque formation assay. A phylogenetic analysis was conducted based on the whole-genome sequencing results. Virus titers were determined, and replication characteristics were analyzed. The efficacy of the isolated strain for in vitro screening of antiviral drugs was validated. Results:A clinical RSV isolate, named hRSV/C-Tan/BJ 202301, was successfully isolated, which could form syncytia in Hep-2 cells. Spherical, filamentous, and irregular virus particles were observed by electron microscopy. Immunofluorescence detection showed green fluorescence in Hep-2 cells, and plaque assay showed round plaques, which were similar to the Long strain in morphology. Genomic sequence analysis showed that it belonged to ON1 genotype. It exhibited similar cell growth kinetics characteristics with the Long strain and could be used for antiviral drug screening in vitro. Conclusions:In this study, one RSV strain is successfully isolated and identified. The biological characteristics and the phylogenetic relationship of this strain reflect the characteristics of the circulating strains in Beijing, which provides experimental material for RSV vaccine development and antiviral drug screening in China.
4.Development and application of an optimized focus-forming assay for quantitation of influenza A virus titer
Jia LI ; Qiaohong CHU ; Lingfang ZHANG ; Xuchang SHAN ; Tangqi WANG ; Ruiwen HAN ; Yujie JIANG ; Donghong WANG ; Baoying HUANG ; Yao DENG ; Wenjie TAN
Chinese Journal of Microbiology and Immunology 2025;45(1):45-52
Objective:To establish and optimize a novel method, focus forming assay (FFA), for quantitation of influenza A virus (FluA) and compare its application performance with traditional plague forming assay (PFA).Methods:The foci chromogenic effects of three peroxidase substrates in immunostaining were compared. The PFA and FFA methods were used to explore FluA incubation times and plaque morphology on 12-well plates, and to determine optimal incubation times and virus adsorption volumes for different FluA subtypes on 96-well plates. The correlation between FFA and PFA was evaluated, and the optimized FFA was applied to the in vitro antiviral efficacy analysis of Favipiravir and neutralization test against different subtypes of FluA. Results:TRUEBLUE substrate was identified as the optimal substrate for foci visualization. Compared with the PFA, the FFA showed improved sensitivity and reduced detection time in FluA titration, and good correlation was shown between the two methods′ results. By replacing the 96-well plate with the 12-well plate for FFA titration of different subtypes of FluA, the detection time was shortened, and the amount of serum samples used could be further reduced by optimizing the virus adsorption volume. The half-maximal effective concentration of favipiravir against influenza viruses assessed by the FFA and PFA methods showed no significant difference, and was consistent with the results obtained from quantitative PCR. Additionally, the focus reduction neutralization test and hemagglutination inhibition assays demonstrated strong correlation in determining antibody titers against FluA in serum neutralization assays.Conclusions:The improved FFA method developed here provides a more efficient experimental tool for FluA titration, antiviral drug screening and broad-spectrum vaccine evaluation.
5.A randomized controlled trial of cognitive behavioral therapy combined with attention training technique on auditory hallucinations of schizophrenia
Xiuli ZHANG ; Xu'en LIU ; Shuo WANG ; Lingfang ZHANG ; Shuanrong LI
Chinese Mental Health Journal 2024;38(3):206-212
Objective:To evaluate the effects of cognitive behavior therapy(CBT)combined with attention training technique(ATT)on auditory hallucinations of schizophrenia.Methods:Sixty schizophrenia patients with au-ditory hallucinations were recruited according to the criteria of International Classification of Diseases,tenth Edition(ICD-10).They were randomly divided into the CBT group(30 cases,26 cases completed)and the CBT+ATT group(30 cases,25 cases completed).The CBT group received only 4 weeks of cognitive behavioral intervention for auditory hallucinations,and the CBT+ATT group received 6 weeks of attention training technology intervention based on cognitive behavioral intervention.At baseline and after 12 weeks,the Psychotic Symptom Rating Scales-Auditory Hallucinations(PSYRATS-AH)and Scale of Social function in Psychosis Inpatients(SSPI)were used to evaluate the auditory hallucinations symptoms and social functioning of the two groups,and the Attentional Control Scale(ACS)was used to evaluate patients'attentional control ability of the CBT+ATT group.Results:After 12 weeks,the differences(post intervention score-baseline score)of PSYRATS-AH total scores[(-6.8±2.2)vs.(-4.2±1.5)],auditory hallucinations'frequency,duration,number of distresses,control of voices were lower in the CBT+ATT group than in the CBT group(Ps<0.05),and the differences of the SSPI total scores[(13.9± 2.9)vs.(11.6±4.2)]and scores of Factor I and Factor Ⅲ were higher in the CBT+ATT group than in the CBT group(Ps<0.05).Conclusion:This study shows cognitive behavioral therapy combined with attention training technique could improve auditory hallucinations and social function more effectively in patients with schizophrenia.
6.Establishment and application of drug use evaluation criteria for roxadustat
Lingfang HUANG ; Shanshan HONG ; Jinhua ZHANG ; Xuhui HUANG
Chinese Journal of Pharmacoepidemiology 2024;33(2):128-136
Objective To establish the drug use evaluation(DUE)standards of roxadustat,and to evaluate its clinical application to promote its rational use.Methods Based on the drug labels,referring to relevant guidelines and expert consensus,the DUE criteria for roxadustat were established through the Delphi method,including items such as drug indications,drug use process and the results of medication.A retrospective study was conducted to evaluate the rationality of cases which included inpatients who used roxadustat for the first time from January 1,2020 to December 31,2022,with a medication period of more than one month in Fujian Provincial Hospital.Results A total of 175 medical records were included,of which 14 records were fully met the clinical application evaluation criteria,and the medication reasonable rate was 8.0%.The unreasonable situation were mainly manifested in the inappropriate clinical outcomes(92.0%),the inappropriate use dosages(52.6%),inappropriate monitoring of adverse reactions(34.9%),inappropriate timing of administration(13.1%),drug interactions(8.7%),inappropriate drug conversion(5.7%),inappropriate efficacy monitoring(4.0%).Conclusion The established DUE standards for roxadustat are scientific,practical and feasible,and the evaluation results show a rate of irrationality in the use of roxadustat in the hospital,and it is necessary to standardize the use of roxadustat in terms of dosage,adverse reaction monitoring,and timing of administration.
7.The mediating role of psychological resilience between work stress and sleep quality in psychiatric nurses
Weina GU ; Lingfang ZHANG ; Zhengjun GUO ; Shichang YANG
Chinese Journal of Modern Nursing 2024;30(12):1638-1642
Objective:To explore the mediating role of psychological resilience between work stress and sleep quality in psychiatric nurses.Methods:From April to August 2023, convenience sampling was used to select 457 psychiatric nurses working in three psychiatric hospitals in Henan Province as the research subjects. The survey was conducted using the General Information Questionnaire, the Chinese version of the Connor-Davidson Resilience Scale (CD-RISC), the Chinese Nursing Stressor Scale (CNSS), and the Pittsburgh Sleep Quality Index (PSQI). Pearson correlation was used to analyze the correlation between psychological resilience, work stress, and sleep quality among psychiatric nurses. SPSS 26.0 software was used to analyze the data, and Process macro program and Bootstrap method were used to test the mediating effect.Results:Among 457 psychiatric nurses, the total scores of the Chinese version of CD-RISC, CNSS, and PSQI were (58.89±13.27), (78.02±14.31), and (8.04±3.46), respectively. Pearson correlation showed that work stress was positively correlated with sleep quality ( r=0.703, P<0.01). In contrast, work stress was negatively correlated with psychological resilience ( r=-0.413, P<0.01), and psychological resilience was negatively correlated with sleep quality ( r=-0.712, P<0.01). Psychological resilience partially mediated the relationship between work stress and sleep quality, with a mediating effect value of 0.051, accounting for 30.0% of the total effect. Conclusions:Psychological resilience partially mediates the relationship between work stress and sleep quality in psychiatric nurses. Nursing managers should attach importance to and enhance the psychological resilience of psychiatric nurses, regularly relieve stress, improve sleep quality, and enhance the physical and mental health of nurses.
8.Effect of night shift work and overweight/obesity on blood pressure of workers in chemical fiber industry
Shuang LIU ; Jianfei WANG ; Tao LI ; Ye ZHU ; Lingfang FENG ; Jiong ZHU ; Jing WANG ; Hailing XIA ; Yiqiu CHEN ; Jiaping LI ; Xing ZHANG ; Zhaoqiang JIANG ; Jianlin LOU
China Occupational Medicine 2023;50(5):529-534
{L-End}Objective To analyze the effects of night shift work and overweight/obesity on blood pressure of workers in chemical fiber industry. {L-End}Methods A total of 1 004 workers of a chemical fiber factory were selected as the study subjects using convenient sampling method, and their blood pressure and body mass index were measured. Multiple linear regression model was used to analyze the relationship between night shift work and blood pressure, and multiple logistic regression was used to assess the independent impact and combined impact of night shifts and overweight/obesity on the risk of hypertension. {L-End}Results Compared with the non-night shift workers, the prevalence of hypertension in night shift workers was increased (5.3% vs 13.0%, P<0.05), with elevated systolic and diastolic blood pressure (both P<0.05). The results of multiple linear regression analysis showed that the systolic blood pressure and diastolic blood pressure of the night shift workers were higher than those of the non-night shift workers (both P<0.05), and the systolic blood pressure and diastolic blood pressure of overweight/obesity workers were higher than those of non-overweight/obesity workers (both P<0.01). The results of multiple logistic regression analysis showed that the risk of hypertension in night shift workers and overweight/obesity workers was higher than that in non-night shift workers and non-overweight/obesity workers [odds ratio (OR) and 95% confidence interval (CI) were 2.49 (1.04-5.99) and 2.65 (1.77-3.95), both P<0.05]. Night shift work and overweight/obesity showed a synergistic effect on blood pressure of workers. Compared to non-overweight/obesity non-night shift workers, overweight/obesity night shift workers had a higher risk of hypertension (OR=4.93, 95%CI: 1.70-14.29, P<0.01). {L-End}Conclusion Night shift work could lead to elevated blood pressure in workers in the chemical fiber industry, which is a potential risk factor for hypertension. The synergistic effect of night shift work and overweight/obesity may contribute to the increased risk of hypertension.
9.Anti-hepatoma Effect of Active Ingredients of Chinese Medicine via Wnt/β-catenin Signaling Pathway: A Review
Yonghua ZHAO ; Yufeng ZHANG ; Yafang FAN ; Nannan LI ; Lingfang LIU
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(21):276-282
Hepatoma is one of the most common malignant tumors of digestive system worldwide, and a main factor leading to cancer-related deaths. Its incidence is increasing year by year, posing a serious threat to human health. Currently, hepatoma is mainly treated by surgical resection, liver transplantation, radiation and drugs, but there are certain adverse reactions and problems of high recurrence rate and low survival rate. Chinese medicine has unique advantages in improving the comprehensive curative effect of hepatoma and reducing adverse reactions. With a variety of active ingredients, Chinese medicine can induce hepatoma cell apoptosis, inhibit the proliferation, migration and reverse multidrug resistance through multiple targets, thus exerting anti-hepatoma effect. It has become an important means for the prevention and treatment of hepatoma as well as a rich resource for anti-hepatoma drug research and development. Wnt/β-catenin signaling pathway, one of the most classical pathways in cancer, is involved in tumor cell proliferation, cell cycle, migration, invasion and tumor angiogenesis. Recently, many studies have reported that the active ingredients of Chinese medicine can play an anti-hepatoma role through this pathway. Therefore, this paper summarized the domestic and foreign literature in recent years, analyzed the relationship between wnt/β-catenin signaling pathway and the specific mechanism of hepatoma occurrence and development, and combed the literature on the effect of flavonoids, terpenoids, alkaloids, polysaccharides and other active ingredients of Chinese medicine on inducing hepatoma cell apoptosis, regulating cell cycle and inhibiting the invasion and metastasis through Wnt/β-catenin signaling pathway. In addition, the paper summarized the research progress of relevant active ingredients of Chinese medicine against hepatoma, to explore their specific mechanism against hepatoma through Wnt/β-catenin signaling pathway, so as to provide theoretical reference for further development of anti-hepatoma drugs.
10.Effect of asbestos exposure on oxidative stress
XIA Hailing ; JIANG Zhaoqiang ; FENG Lingfang ; YU Min ; ZHANG Min ; CHEN Junqiang ; ZHANG Xing ; LOU Jianlin
Journal of Preventive Medicine 2022;34(1):1-6
Objective :
To examine the effect of asbestos exposure on oxidative stress, so as to provide insights into the elucidation of pathogenesis and management of asbestos-related diseases.
Methods :
Totally 245 subjects were recruited from an asbestos manufacturing area in Zhejiang Province, and their gender, age and history of asbestos exposure were collected through a questionnaire survey. The serum levels of 8-hydroxy-2'deoxyguanosine ( 8-OHdG ), glutathione ( GSH ), malondialdehyde ( MDA ), superoxide dismutase ( SOD ) and total antioxidative capacity ( TAOC ) were measured using an enzyme-linked immunosorbent assay ( ELISA ), and the levels of catalase ( CAT ), peroxiredoxin 2 ( PRX2 ), SOD1, SOD2 and thioredoxin-1 ( TRX1 ) were detected in peripheral white blood cells ( WBCs ) using a liquid-chip assay. Multivariable linear regression analysis was performed to identify the association between asbestos exposure and oxidative stress parameters.
Results :
There were 50 subjects without a history of asbestos exposure (unexposed group), 102 subjects with asbestos exposure for less than 10 years ( AE<10-year group ) and 93 subjects with asbestos exposure for 10 years and more ( AE≥10-year group ). No significant differences were found among the three groups in terms of age, gender, proportion of smokers or proportion of alcohol consumers ( P>0.05 ). Significantly higher 8-OHdG and MDA in serum, and higher PRX2 in peripheral WBCs were detected in the AE≥10-year group than in the unexposed group ( P<0.05 ); lower GSH and TAOC in serum, and lower CAT in peripheral WBCs were detected in the AE≥10-year group than in the unexposed group ( P<0.05 ); higher 8-OHdG and MDA in serum, and higher PRX2 in peripheral WBCs were detected in the AE≥10-year group than in the AE<10-year group ( P<0.05 ). Multivariable linear regression analysis showed that asbestos exposure significantly correlated with 8-OHdG, MDA and TAOC in serum, and CAT and PRX2 in peripheral WBCs ( P<0.05 ).
Conclusion
Asbestos exposure may induce the oxidative stress damage, suggesting that oxidative stress may be involved in asbestos-related diseases.


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