1.Effect of Yifei Jianpi Prescription on Lipopolysaccharide-induced Lung Immune Inflammatory Response in Rats Based on STAT1/IRF3 Pathway
Hongjuan YANG ; Yaru YANG ; Yujie YANG ; Zhongbo ZHU ; Quan MA ; Yanlin WU ; Hongmei LI ; Xuhui ZHANG ; Xiping LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):146-155
ObjectiveTo observe the effect of Yifei Jianpi prescription on the of signal transducer and activator of transcription protein 1 (STAT1)/interferon regulatory factor 3 (IRF3) signaling pathway in a pneumonia model induced by lipopolysaccharide (LPS) and to explore the mechanism of Yifei Jianpi prescription in improving lung immune and inflammatory responses. MethodsSixty male SPF SD rats were used in this study. Ten rats were randomly assigned to the normal control group, and the remaining 50 were instilled with LPS in the trachea to establish a pneumonia model. After successful modeling, the rats were randomly divided into the model group, dexamethasone group (0.5 mg·kg-1), and Yifei Jianpi prescription high-dose (12 mg·kg-1), medium-dose (6 mg·kg-1), and low-dose (3 mg·kg-1) groups, with 10 rats in each group. Treatment was administered once daily, and the normal control and model groups received the same volume of normal saline. After 14 days, flow cytometry was used to detect the classification of whole blood lymphocytes. Enzyme-linked immunosorbent assay (ELISA) was used to measure serum levels of immunoglobulin G (IgG), immunoglobulin A (IgA), immunoglobulin M (IgM), and the content of tumor necrosis factor-α (TNF-α), interleukin-8 (IL-8), interleukin-6 (IL-6), and interleukin-10 (IL-10) in alveolar lavage fluid (BALF). Hematoxylin-eosin (HE) staining was used to observe lung tissue pathology and score the damage. Thymus weight, spleen weight, and wet-to-dry weight ratio (W/D) were recorded. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of STAT1, IRF3, IL-6, and interferon-alpha (IFN-α) in lung tissues, while Western blot was performed to assess the protein expression of STAT1, IRF3, IL-6, and IFN-α. ResultsCompared with the normal control group, the model group showed significantly increased proportion of B lymphocytes in peripheral blood, decreased proportions of NK cells and CD4+/CD8+ (P<0.05, P<0.01), decreased serum levels of IgG and IgA, significantly increased IgM levels (P<0.01), significantly elevated content of TNF-α, IL-6, and IL-8 in BALF, and significantly decreased IL-10 levels (P<0.01). Lung tissue damage was evident, with significant increases in thymus and spleen weights and a higher W/D ratio (P<0.01). The mRNA and protein expression of STAT1, IRF3, IFN-α, and IL-6 in lung tissues was significantly upregulated (P<0.05,P<0.01). Compared with the model group, the Yifei Jianpi prescription groups showed significantly reduced proportions of B lymphocytes in peripheral blood, increased proportions of NK cells and CD4+/CD8+ ratios (P<0.05, P<0.01), significantly increased serum levels of IgG and IgA, significantly decreased IgM levels (P<0.05, P<0.01), significantly reduced levels of TNF-α, IL-6, and IL-8 in BALF, and significantly increased IL-10 levels (P<0.01). Lung tissue damage was alleviated, thymus and spleen weights were significantly reduced, and the W/D ratio was markedly decreased (P<0.01). The mRNA and protein expression of STAT1, IRF3, IFN-α, and IL-6 in lung tissues was significantly downregulated (P<0.05, P<0.01). ConclusionYifei Jianpi prescription can alleviate lung tissue damage and improve immune and inflammatory responses in LPS-induced pneumonia rats. The mechanism may be related to the inhibition of STAT1/IRF3 signaling pathway activation.
2.Progress of clinical detection methodology of varicella vaccine serum antibody
Yaru QUAN ; Yanli DAI ; Changgui LI ; Kangwei XU ; Jizong JIA
Chinese Journal of Laboratory Medicine 2025;48(6):785-790
Varicella is a common infectious disease caused by varicella-zoster virus (VZV), with high transmissibility and pathogenicity, and vaccination is a better preventive measure. The clinical diagnosis of varicella, the clinical immunogenicity evaluation of varicella vaccine, the continuous monitoring of the immune effect after marketing and the epidemiological investigation all require the detection of the antibody level of VZV in patients. Fluorescent antibody to membrane antigen method (FAMA) is the gold standard for detection of VZV antibody in serum, possessing advantages of high sensitivity and good specificity, playing an important role in detection of VZV antibody. However, the defects of FAMA operation complexity and low throughput are also troubling the clinical testing personnels. This article reviews research progress both at home and abroad on the technology of VZV serum antibody detection. The advantages and limitations of the current detection methods are discussed and the future development direction are highlighted on the dectection of VZV serum antibody IgG.
3.Development of an endotoxin detection method for mRNA vaccines based on lipid nanoparticle delivery system
Jiaru CAI ; Shaoyi CHEN ; Shuang LI ; Juan LI ; Yucai PENG ; Yaru QUAN
Chinese Journal of Microbiology and Immunology 2025;45(11):958-964
Objective:To develop a bacterial endotoxin detection method suitable for mRNA vaccines based on lipid nanoparticle(LNP)delivery system.Methods:The gel clot limit test outlined in the third section of the Chinese Pharmacopoeia(2020 edition)was utilized. Taking the human herpesvirus type 2 mRNA vaccine as the detection object,the optimal conditions for bacterial endotoxin detection were established by evaluating sample dilution factors,diluents,and other interfering elements. The results were validated using the recombinant C factor method. Finally,the applicability of the established method was validated through testing the respiratory syncytial virus mRNA vaccine and the shingles mRNA vaccine.Results:Increasing the sample dilution factor to 480 times with a magnesium-containing buffer and using 0.06 EU/ml tachypleus amebocyte lysate effectively mitigated the interference caused by LNP on the test results of the human herpesvirus type 2 mRNA vaccine,the respiratory syncytial virus mRNA vaccine,and the shingles mRNA vaccine. The results from recombinant C factor method and the gel clot limit method were consistent.Conclusion:A bacterial endotoxin gel clot limit detection method tailored for human herpesvirus type 2 mRNA vaccine,respiratory syncytial virus mRNA vaccine,and shingles mRNA vaccine is successfully developed based on existing protocols in the Chinese Pharmacopoeia(2020 edition). This study offers insights for the detection of bacterial endotoxin in LNP-based biological products.
4.Development of an endotoxin detection method for mRNA vaccines based on lipid nanoparticle delivery system
Jiaru CAI ; Shaoyi CHEN ; Shuang LI ; Juan LI ; Yucai PENG ; Yaru QUAN
Chinese Journal of Microbiology and Immunology 2025;45(11):958-964
Objective:To develop a bacterial endotoxin detection method suitable for mRNA vaccines based on lipid nanoparticle(LNP)delivery system.Methods:The gel clot limit test outlined in the third section of the Chinese Pharmacopoeia(2020 edition)was utilized. Taking the human herpesvirus type 2 mRNA vaccine as the detection object,the optimal conditions for bacterial endotoxin detection were established by evaluating sample dilution factors,diluents,and other interfering elements. The results were validated using the recombinant C factor method. Finally,the applicability of the established method was validated through testing the respiratory syncytial virus mRNA vaccine and the shingles mRNA vaccine.Results:Increasing the sample dilution factor to 480 times with a magnesium-containing buffer and using 0.06 EU/ml tachypleus amebocyte lysate effectively mitigated the interference caused by LNP on the test results of the human herpesvirus type 2 mRNA vaccine,the respiratory syncytial virus mRNA vaccine,and the shingles mRNA vaccine. The results from recombinant C factor method and the gel clot limit method were consistent.Conclusion:A bacterial endotoxin gel clot limit detection method tailored for human herpesvirus type 2 mRNA vaccine,respiratory syncytial virus mRNA vaccine,and shingles mRNA vaccine is successfully developed based on existing protocols in the Chinese Pharmacopoeia(2020 edition). This study offers insights for the detection of bacterial endotoxin in LNP-based biological products.
5.Progress of clinical detection methodology of varicella vaccine serum antibody
Yaru QUAN ; Yanli DAI ; Changgui LI ; Kangwei XU ; Jizong JIA
Chinese Journal of Laboratory Medicine 2025;48(6):785-790
Varicella is a common infectious disease caused by varicella-zoster virus (VZV), with high transmissibility and pathogenicity, and vaccination is a better preventive measure. The clinical diagnosis of varicella, the clinical immunogenicity evaluation of varicella vaccine, the continuous monitoring of the immune effect after marketing and the epidemiological investigation all require the detection of the antibody level of VZV in patients. Fluorescent antibody to membrane antigen method (FAMA) is the gold standard for detection of VZV antibody in serum, possessing advantages of high sensitivity and good specificity, playing an important role in detection of VZV antibody. However, the defects of FAMA operation complexity and low throughput are also troubling the clinical testing personnels. This article reviews research progress both at home and abroad on the technology of VZV serum antibody detection. The advantages and limitations of the current detection methods are discussed and the future development direction are highlighted on the dectection of VZV serum antibody IgG.
6.Research progess in respiratory syncytial virus receptors
Shuyan LI ; Huijie YANG ; Yaru QUAN ; Changgui LI
Chinese Journal of Microbiology and Immunology 2024;44(2):183-188
Respiratory syncytial virus (RSV) is one of the major pathogens of acute respiratory infections, becoming a huge global burden. Virus-receptor interactions play a key role in the pathogenesis of RSV infection. The distribution of receptors influences the cellular and the tissue tropism of RSV as well as the viral replication and proliferation in the host. However, the RSV receptors are currently unknown, which is one of the reasons that hinders the development of RSV vaccines and therapeutic drugs. In this study, the existing RSV receptors are reviewed with the hope to provide ideas for the research of RSV vaccines and therapeutic drugs.
7.Correlation between 23 metals and metalloids elements in the urine and thyroid function of traffic police
Yaru WANG ; Chao QUAN ; Tian XYU ; Shuxia LIU ; Lizhao YANG ; Ling ZHANG ; Xiayun DAI
Journal of Public Health and Preventive Medicine 2024;35(2):25-29
Objective To investigate the correlation between 23 metals and metalloids elements in the urine and thyroid function indicators in the blood of traffic police. Methods A cross-sectional study was performed to assess the effects of 23 metals and metalloids elements in the urine on blood thyroid function indicators in 166 traffic policemen (122 field staff and 44 internal staff) in Wuhan, Hubei Province. Each subject received an occupational health examination. Results After multiple corrections for false detection rates, in the polymetallic model, the levels of urinary manganese and urinary uranium were positively correlated with the levels of thyroid peroxidase antibody (TPOAb) in the blood (β = 66.57, 95% CI 2.92-130.22, P = 0.040 and β = 62.43, 95% CI 14.37-110.49, P = 0.011), and the level of urinary uranium was positively correlated with thyroid stimulating hormone (TSH) levels in the blood (β = 6.20, 95% CI 2.68-9.72 , P = 0.001). Urinary uranium level was negatively correlated with free thyroxine level in the blood (FT4) (β = -2.03, 95 % CI (-3.67 )- (-0.39), P = 0.015), and urinary lead level was negatively correlated with blood TSH level (β = -4.59, 95% CI (-8.67) - (-0.51), P = 0.027). Conclusion Manganese exposure is related to the increase of TPOAb level in blood, uranium exposure is related to the increase of TPOAb and TSH levels and the decrease of FT4 level in blood, and lead exposure is related to the decrease of TSH level in blood, suggesting that more attention should be paid to the effects of heavy metals on the thyroid of traffic police.
8.Comparison of two detection methods for antibody to varicella-zoster virus
Chinese Journal of Biologicals 2024;37(12):1490-1494
Objective To compare the detection ability of the developed enzyme-linked immunosorbent assay(ELISA) and the fluorescent antibody-to-membrane antigen(FAMA) assay for the IgG antibody to varicella-zoster virus(VZV), and to provide a reference for the evaluation of vaccine immune effect and the study of auxiliary immunization strategy.Methods Glycoproteins gE/gB were used as coating antigen to develop an ELISA method for quantitative detection of VZV IgG antibody. The developed ELISA and FAMA assay were used to detect VZV IgG antibody levels in 2 332 varicella vaccine serum samples(1 166 samples before and after immunization respectively), and the results of the two methods were compared and analyzed.Results The total positive rates detected by ELISA and FAMA assay were 88% and 89. 8%, the positive rates of serum samples before immunization were 76. 3% and 79. 6%, and the positive rates after immunization were 99. 7% and100%, respectively. The total coincidence rate of the two methods was 95. 8%, and Kappa value was 0. 789. The correlation coefficient(R~2) between antibody concentration detected by ELISA and antibody titer detected by FAMA assay was 0. 705.Conclusion The developed ELISA method has high consistency and correlation with FAMA assay, which can be used to evaluate the immune effect of vaccines and study immunization strategies.
9.Analysis of the etiological features and clinical characteristics of herpes zoster cases under 20 years old in Beijing City from 2017 to 2021
Jingbin PAN ; Tao ZHOU ; Luodan SUO ; Liyong YUAN ; Yaru QUAN ; Haihong WANG ; Xinghui PENG ; Tao WANG ; Zonglong ZHU ; Yanfei WANG ; Dan ZHAO ; Li LU
Chinese Journal of Preventive Medicine 2024;58(3):285-291
Objective:To analyze the etiological features and clinical characteristics of herpes zoster cases under 20 years old in Beijing City from 2017 to 2021.Methods:Herpes zoster cases were collected from a surveillance system in Beijing City from December 2017 to April 2021. The cases included individuals under 20 years old from seven sentinel hospitals located in two districts (Miyun District and Changping District). The basic information, the rash date of rash onset and the location and number of lesions were investigated at the first visit to the hospital, and the lesion swab samples were collected for laboratory testing. A telephone follow-up was conducted 21 days after the onset of the rash to investigate the degree of pain, duration of the rash and duration of pain. The individuals who still experienced neuralgia were further investigated for their pain condition at 90 days after the onset of the rash, to discover cases with postherpetic neuralgia. DNA was extracted from the rash fluid, and the ORF62 gene region was amplified and sequenced to obtain the viral sequence. The wild-type strain or chickenpox vaccine strain was identified by using sequence alignment, and the clinical characteristics of cases with different varicella vaccinations were compared.Results:A total of 78 herpes zoster cases under 20 years old were investigated during 2017-2021 in Beijing City, and 61 cases completed the follow-up survey. The age range of 61 cases was 1.83 to 20.54 years with a median age of 17.50 years. There were 36 males (59.02%) and 25 females (40.98%). Among them, there were 29 cases with the chickenpox vaccine immunization history (18 cases with one dose, 5 cases with two doses and 6 cases with unknown doses), 13 cases with no vaccination history and 19 cases with unknown vaccination history. Among the 78 cases, the herpetic fluid samples of 64 cases were positive for VZV, including 62 cases identified as wild-type strains and two cases as vaccine strains. The two vaccine strain cases were both 2-year-old girls who had received one dose of varicella vaccine and developed herpes zoster 3 months and 13 months after vaccination. Among the 29 cases with chickenpox vaccine immunization history, the majority had 10 to 49 lesions, accounting for 58.62% (17 cases). The trunk was the most common site of lesions, accounting for 44.83% (13 cases). About 51.72% (15 cases) reported "no or mild" pain intensity. The median ( Q1, Q3) scores for the worst pain, duration of pain and the time to crusting of lesions in the herpes zoster cases were 3 (1.5, 5) points, 10 (1.5, 12.5) days and 10 (6.5, 13) days, respectively. There was no statistically significant difference in the constituent ratio of the location of lesions, number of lesions and pain degree among the cases with vaccination history, without vaccination history and with unknown vaccination history ( P>0.05). There was also no statistically significant difference in the distribution of pain score, duration of lesions and duration of pain across the three groups ( P>0.05). Conclusion:Wild strains are the predominant pathogens in herpes zoster cases under 20 years old in Beijing City during 2017-2021. The varicella vaccination has no significant impact on the clinical manifestations of herpes zoster cases.
10.Analysis of the etiological features and clinical characteristics of herpes zoster cases under 20 years old in Beijing City from 2017 to 2021
Jingbin PAN ; Tao ZHOU ; Luodan SUO ; Liyong YUAN ; Yaru QUAN ; Haihong WANG ; Xinghui PENG ; Tao WANG ; Zonglong ZHU ; Yanfei WANG ; Dan ZHAO ; Li LU
Chinese Journal of Preventive Medicine 2024;58(3):285-291
Objective:To analyze the etiological features and clinical characteristics of herpes zoster cases under 20 years old in Beijing City from 2017 to 2021.Methods:Herpes zoster cases were collected from a surveillance system in Beijing City from December 2017 to April 2021. The cases included individuals under 20 years old from seven sentinel hospitals located in two districts (Miyun District and Changping District). The basic information, the rash date of rash onset and the location and number of lesions were investigated at the first visit to the hospital, and the lesion swab samples were collected for laboratory testing. A telephone follow-up was conducted 21 days after the onset of the rash to investigate the degree of pain, duration of the rash and duration of pain. The individuals who still experienced neuralgia were further investigated for their pain condition at 90 days after the onset of the rash, to discover cases with postherpetic neuralgia. DNA was extracted from the rash fluid, and the ORF62 gene region was amplified and sequenced to obtain the viral sequence. The wild-type strain or chickenpox vaccine strain was identified by using sequence alignment, and the clinical characteristics of cases with different varicella vaccinations were compared.Results:A total of 78 herpes zoster cases under 20 years old were investigated during 2017-2021 in Beijing City, and 61 cases completed the follow-up survey. The age range of 61 cases was 1.83 to 20.54 years with a median age of 17.50 years. There were 36 males (59.02%) and 25 females (40.98%). Among them, there were 29 cases with the chickenpox vaccine immunization history (18 cases with one dose, 5 cases with two doses and 6 cases with unknown doses), 13 cases with no vaccination history and 19 cases with unknown vaccination history. Among the 78 cases, the herpetic fluid samples of 64 cases were positive for VZV, including 62 cases identified as wild-type strains and two cases as vaccine strains. The two vaccine strain cases were both 2-year-old girls who had received one dose of varicella vaccine and developed herpes zoster 3 months and 13 months after vaccination. Among the 29 cases with chickenpox vaccine immunization history, the majority had 10 to 49 lesions, accounting for 58.62% (17 cases). The trunk was the most common site of lesions, accounting for 44.83% (13 cases). About 51.72% (15 cases) reported "no or mild" pain intensity. The median ( Q1, Q3) scores for the worst pain, duration of pain and the time to crusting of lesions in the herpes zoster cases were 3 (1.5, 5) points, 10 (1.5, 12.5) days and 10 (6.5, 13) days, respectively. There was no statistically significant difference in the constituent ratio of the location of lesions, number of lesions and pain degree among the cases with vaccination history, without vaccination history and with unknown vaccination history ( P>0.05). There was also no statistically significant difference in the distribution of pain score, duration of lesions and duration of pain across the three groups ( P>0.05). Conclusion:Wild strains are the predominant pathogens in herpes zoster cases under 20 years old in Beijing City during 2017-2021. The varicella vaccination has no significant impact on the clinical manifestations of herpes zoster cases.


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