1.Detection of serum antibody levels after inoculation of an inactivated SARS-CoV-2 vaccine
XU Nani XU Nani ; HU Xiaowei HU Xiaowei ; LI Haiyan LI Haiyan ; ZHENG Lin ZHENG Lin ; KONG Qingxin KONG Qingxin ; QIN Pan QIN Pan ; LI Jing LI Jing ; YE Fei YE Fei
Journal of Preventive Medicine 2022;34(1):33-37
Objective :
To investigate the serum levels of antibodies against SARS-CoV-2 after inoculation of an inactivated SARS-CoV-2 vaccine, so as to provide insights into the evaluation of the vaccine immunogenicity.
Methods :
In this single-arm Objective performance criteria trial, residents aged 18 to 59 years and inoculated with an inactivated SARS-CoV-2 vaccine in Xihu District, Hangzhou City from October to December of 2020 were selected using a cluster sampling method. Blood samples were collected prior to inoculation, 14 and 28 days post-inoculation of the first dose, and 28 days post-inoculation of the second dose. Serum levels of anti-SARS-CoV-2 IgM and IgG antibodies were detected using the magnetic particle-based chemiluminescence immunoassay. The seroconversion of antibodies and dynamic changes of antibody levels were analyzed.
Results :
Totally 310 participants were enrolled, including 133 subjects on day 14 post-inoculation of the first dose, 97 subjects on day 28 post-inoculation of the first dose and 254 subjects on day 28 post-inoculation of the second dose. The seroconversion rates of anti-SARS-CoV-2 IgG antibody were 6.02%, 28.87% and 98.43%, and the median IgG antibody levels were 1.76 ( interquartile range, 3.25 ), 5.69 ( 9.95 ) and 52.05 ( 47.60 ) AU/mL ( P<0.05 ), respectively, while the seroconversion rates of anti-SARS-CoV-2 IgM antibody were 9.02%, 11.34% and 12.99%, and the median IgG antibody levels were 1.89 ( 3.28 ), 2.06 ( 4.71 ) and 2.65 ( 4.01 ) AU/mL ( P>0.05 ), respectively. In addition, higher serum levels of anti-SARS-CoV-2 IgG and IgM antibodies were detected post-inoculation relative to pre-inoculation ( P<0.05 ), and higher serum IgG antibody levels were found in subjects aged 18 to 39 years than in those aged 40 to 59 years ( P<0.05 ).
Conclusions
Inoculation of two doses of the inactivated SARS-CoV-2 vaccine achieves a high immunogenicity among residents aged 18 to 59 years 28 days post-inoculation, and the anti-SARS-CoV-2 IgM antibody is detectable in some residents following inoculation of the first dose.
2.Simultaneous Determination of Chromium Ⅲ, Chromium Ⅵin Leather and Cloth by Ion Chromatography with On-line Sample Pretreatment Column-Switching Technique
Jie HE ; Jiasheng YU ; Zhongping HUANG ; Zuoyi ZHU ; Muhua WANG ; Nani WANG ; Jiajie ZHANG ; Peimin ZHANG ; Gang LI ; Yan ZHU
Chinese Journal of Analytical Chemistry 2014;(8):1190-1195
A new analytical method has been developed for the simultaneous determination of CrⅢand CrⅥusing on-line sample pretreatment valve-switching ion chromatography. The organic matrix in leather was removed by using a reverse-phase column as the pretreatment column. Before injection, EDTA was added into sample solution to react with the CrⅢto form anion which could absorb visible light strongly. After injection, the ions separated by the pretreatment column were received in a collection loop. Then the ions were delivered into an analytical column and separated. CrⅥ then was derived with the derivatization reagent 1, 5-diphenylcarbazide ( DPC) , and detected together with CrⅢ-EDTA complex by a UV-Vis detector. Under the optimum conditions, the linear range of the method for CrⅢ and CrⅥ was 0. 3-10 mg/L (r=0. 9991) and 0. 05-2 mg/L ( r = 0. 9992 ), whereas detection limits ( S/N = 3 ) were 80. 78 μg/L and 6. 67 μg/L, respectively. The recoveries were in the range of 88. 7%-108. 5% with the relative standard deviations for retention time and peak area less than 3%. The method could be applied to determine CrⅢ and CrⅥ in leather and cloth effectively and quickly.
3.Predicting the Prognosis and Immunotherapeutic Response of Triple-Negative Breast Cancer by Constructing a Prognostic Model Based on CD8+T Cell-Related Immune Genes
Li NANI ; Qiu XIAOTING ; Xue JINGSONG ; Yi LIMU ; Chen MULAN ; Huang ZHIJIAN
Biomedical and Environmental Sciences 2024;37(6):581-593
Objective Triple-negative breast cancer(TNBC)poses a significant challenge for treatment efficacy.CD8+T cells,which are pivotal immune cells,can be effectively analyzed for differential gene expression across diverse cell populations owing to rapid advancements in sequencing technology.By leveraging these genes,our objective was to develop a prognostic model that accurately predicts the prognosis of patients with TNBC and their responsiveness to immunotherapy. Methods Sample information and clinical data of TNBC were sourced from The Cancer Genome Atlas and METABRIC databases.In the initial stage,we identified 67 differentially expressed genes associated with immune response in CD8+T cells.Subsequently,we narrowed our focus to three key genes,namely CXCL13,GBP2,and GZMB,which were used to construct a prognostic model.The accuracy of the model was assessed using the validation set data and receiver operating characteristic(ROC)curves.Furthermore,we employed various methods,including Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway,immune infiltration,and correlation analyses with CD274(PD-L1)to explore the model's predictive efficacy in immunotherapeutic responses.Additionally,we investigated the potential underlying biological pathways that contribute to divergent treatment responses. Results We successfully developed a model capable of predicting the prognosis of patients with TNBC.The areas under the curve(AUC)values for the 1-,3-,and 5-year survival predictions were 0.618,0.652,and 0.826,respectively.Employing this risk model,we stratified the samples into high-and low-risk groups.Through KEGG enrichment analysis,we observed that the high-risk group predominantly exhibited enrichment in metabolism-related pathways such as drug and chlorophyll metabolism,whereas the low-risk group demonstrated significant enrichment in cytokine pathways.Furthermore,immune landscape analysis revealed noteworthy variations between(PD-L1)expression and risk scores, Conclusion Our study demonstrates the potential of CXCL13,GBP2,and GZMB as prognostic indicators of clinical outcomes and immunotherapy responses in patients with TNBC.These findings provide valuable insights and novel avenues for developing immunotherapeutic approaches targeting TNBC.
4.Study on specific antibodies in patients infected with SARS-CoV-2 and recipients of inactivated vaccine
Nani XU ; Jianhua YU ; Xiaowei HU ; Haiyan LI ; Lin ZHENG ; Qingxin KONG ; Fei YE ; Jing LI ; Pan QIN
Chinese Journal of Microbiology and Immunology 2022;42(1):16-22
Objective:To detect the serum levels of SARS-CoV-2-specific IgM and IgG antibodies in patients infected with SARS-CoV-2 and recipients of inactivated vaccine in different periods for understanding their variation patterns in vivo. Methods:Chemiluminescence immunoassay was used to detect the levels of SARS-CoV-2-specific IgM and IgG antibodies in 144 serum samples of 44 COVID-19 patients, 381 serum samples of 118 asymptomatic infected cases and 398 serum samples of 273 inactivated vaccine recipients collected at different periods. The results were statistically analyzed together with basic characteristics and vaccination status.Results:The positive rates of IgM antibody in COVID-19 patients, asymptomatic infected cases and inactivated vaccine recipients were 52.27% (23/44), 23.73% (28/118) and 14.29% (39/273). The positive rate of IgM antibody was higher in COVID-19 patients than in asymptomatic infected cases and vaccine recipients (χ 2=12.106, P=0.001; χ 2=34.755, P<0.001). The positive rates of IgG antibody in the three populations were 100.00% (44/44), 97.46% (115/118) and 98.81% (166/168), and the differences were not statistically significant (χ 2=2.944, P=0.229). In COVID-19 patients, the concentration of IgM antibody in <40 years old group was lower than that in ≥40 years old group (Waldχ 2=6.609, P=0.010), and the concentration of IgG antibody in patients with vaccination was higher than that in patients without vaccination (Waldχ 2=12.402, P<0.001). In asymptomatic infected cases, the concentration of IgG antibody was higher in people with vaccination than in those without vaccination (Waldχ 2=4.530, P=0.033). In SARS-CoV-2 vaccine recipients, the concentration of IgG antibody in <40 years old group was higher than that in ≥40 years old group (Waldχ 2=9.565, P=0.002). Dynamic analysis of antibody levels showed that from week 1 to week 9, the concentrations of IgM and IgG antibodies in COVID-19 patients were higher than those in asymptomatic infected cases and vaccine recipients. Conclusions:The concentrations of IgM and IgG antibodies in COVID-19 patients were higher than those in asymptomatic infected cases and inactivated vaccine recipients. COVID-19 patients aged ≥40 years had higher level of IgM antibody. COVID-19 patients and asymptomatic infected cases who had received vaccination had higher concentration of IgG antibody. Inactivated vaccine showed good immunogenicity after whole course of immunization, and the IgG antibody level in <40 years old group was higher.
5.Breeding of transgenic mice expressing human tau isoform with P301L mutation and identification of homozygous transgenic mice.
Yan-yan WANG ; Ru-zhui CHEN ; Xiao-nani ZHU ; Jing LIU ; Zhi-hui LI ; Xiu-juan LIU ; Zhi-hui LI ; Xin NA ; Shan-shan LIANG ; Guo-guang QIU ; Wei ZHANG ; Hai WANG ; Xue-lan WANG
Chinese Journal of Applied Physiology 2012;28(3):221-224
OBJECTIVETo establish homozygous transgenic mouse strain expressing human tau isoform with P301L mutation.
METHODSFive transgenic mice expressing human tau isoform with P301L mutation were obtained by microinjection into male nuclei. Homozygote and hemizygote were identified by PCR and real-time fluorescent quantitative PCR.
RESULTSNinety five homozygous transgenic mice were selected, and the results indicated that homozygous transgenic mice were superior to hemizygote in simulating the changes of biological characteristics.
CONCLUSIONExogenous gene tau is able to stably transmit to next generation and the combination of SYBR Green real-time fluorescent quantitative PCR with the traditional mating is a fast, reliable and economical way to screen homozygous and hemizygous transgenic mice.
Animals ; Female ; Homozygote ; Humans ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Microinjections ; Mutation ; tau Proteins ; genetics
6.Simultaneous Determination of 4 Triglyceride Anti-tumor Components in Coix lacryma Seed Oil by Quatitative Analysis of Multi-components by Single Marker
Renjie WU ; Pingcui XU ; Dan SHOU ; Gonghua LI ; Qingshan LAN ; Nani WANG
China Pharmacy 2019;30(10):1375-1380
OBJECTIVE: To establish a method for simultaneous determination of 4 triglyceride anti-tumor components in Coix lacryma seed oil. METHODS: HPLC-ELSD was adopted. The determination was performed on Inertsil ODS-3 C18 column with mobile phase consisted of acetonitrile-isopropanol (57 ∶ 43, V/V) at the flow rate of 1.0 mL/min. The column temperature was 30 ℃, and the sample size was 10 μL. Evaporative light scattering detector was used, the drift tube temperature was 70 ℃, and the gas flow rate was 2 L/min. Using glycerol trioleateas internal standard, relative correction factors (RCF) of linolein trilinolein, 1,2-linoleic acid-3-palmitic acid glyceride and 1-palmitic acid-2-oleic acid-3-linoleic acid glyceride were calculated respectively. The contents of above 3 components in C. lacryma seed oil were calculated by RCF. The contents of 4 components in C. lacryma seed oil were determined by external standard. The results of content determination by quatitative analysis of multi-components by single marker (QAMS) were compared with external standard method. RESULTS: The linear ranges were 0.15-4.50 μg for linolein trilinolein, 0.15-4.50 μg for 1,2-linoleic acid-3-palmitate, 0.35-10.50 μg for 1-palmitic acid-2-oleic acid-3-linoleic acid glyceride, 0.35-10.50 μg for glycerol trioleate (r≥0.999 5). The limits of quantification were 0.13, 0.06, 0.07, 0.12 μg. The limits of detection were 0.04, 0.02, 0.02, 0.03 μg, respectively. RSDs of precision, stability, and repeatability tests were less than 2.0%(n=6). The average recoveries were 95.43%-102.67%(RSD<2.0%, n=6). Average RCFs of linolein trilinolein, 1,2- linoleic acid-3-palmitic acid glyceride and 1-palmitic acid-2- oleic acid-3-linoleic acid glyceride were 0.31, 0.88, and 1.21, respectively. RCFs reproducibility was perfect under different experiment conditions. There was no significant difference in results of content determination between QAMS and external standard method (P>0.05). CONCLUSIONS: The method is simple, rapid, accurate and reliable. It is used for simultaneous determination of linolein trilinolein, 1, 2-linoleic acid-3-palmitate, 1-palmitic acid-2-oleic acid-3-linoleic acid glyceride and glycerol trioleateas in C. lacryma seed oil.