1.Prokaryotic expression, purification and immunogenicity of SARS-CoV-2 omicron variant nucleocapsid protein.
Zewen TU ; Quansheng WANG ; Shiguo LIU ; Haosen LIU ; Chunyan ZENG ; Juanjuan XIE ; Mingzhi LI ; Jingcai LI ; Min WANG ; Shiqi WENG ; Lumei KANG ; Lingbao KONG
Chinese Journal of Cellular and Molecular Immunology 2025;41(8):735-743
Objective The study aims to investigate the immunological functions of the nucleocapsid (N) protein of the novel coronavirus Omicron (BA.1, BA.2) and evaluate the differences among different N proteins of mutant strains in immunogenicity. Methods By aligning sequences, the mutation sites of the Omicron (BA.1, BA.2) N protein relative to prototype strain of the novel coronavirus (Wuhan-Hu-1) were determined. The pET-28a-N-Wuhan-Hu-1 plasmid was used as template to construct pET-28a-BA.1/BA.2-N through single point mutation or homologous recombination. The three kinds of N protein were expressed in prokaryotic system, purified through Ni-NTA affinity chromatography, and then immunized into mice. The titer and reactivity of the polyclonal antibody, as well as the expression level of IL-1β and IFN-γ in mouse spleen cells, were detected using indirect ELISA and Western blot assay. Results The constructed prokaryotic expression plasmids were successfully used to express the Wuhan-Hu-1 N, BA.1 N, and BA.2 N proteins in E.coli BL21(DE3) at 37 DegreesCelsius for 4 hours. The indirect ELISA test showed that the titers of polyclonal antibody prepared by three N proteins were all 1:51 200. All three N proteins can increase the expression of IFN-γ and IL-1β cytokines, but the effect of Omicron N protein in activing two cytokines was more obvious than that of Wuhan-Hu-1 N protein. Conclusion The study obtained three new coronavirus N proteins and polyclonal antibodies, and confirmed that mutations in the amino acid sites of the N protein can affect its immunogenicity. This provides a basis for developing rapid diagnostic methods targeting N protein of different novel coronavirus variants.
Animals
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Mice
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SARS-CoV-2/genetics*
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Coronavirus Nucleocapsid Proteins/immunology*
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Nucleocapsid Proteins/isolation & purification*
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COVID-19/immunology*
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Antibodies, Viral/immunology*
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Mice, Inbred BALB C
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Interferon-gamma/metabolism*
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Interleukin-1beta/metabolism*
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Female
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Escherichia coli/metabolism*
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Mutation
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Humans
2.Clinical characteristics of postoperative pneumonia patients undergoing different types of surgeries and distribution of onset time
Ran XIN ; Lei QIAO ; Shuangfeng SUN ; Yingying PAN ; Juanjuan KONG ; Hongyu WANG ; Ying YAN
Chinese Journal of Nosocomiology 2025;35(20):3074-3078
OBJECTIVE To explore the time windows for postoperative pneumonia in patients undergoing different surgeries,providing evidence-based references for optimizing infection monitoring and prevention and control strategies.METHODS Sociodemographic characteristics,clinical information and surgical details of 263 patients with postoperative pneumonia from four different types of medical institutions between Jan.2019 and Dec.2024 were retrospectively collected.The time windows for postoperative pneumonia in patients undergoing different surgeries were analyzed.RESULTS There were no statistically significant differences in the time windows for post-operative pneumonia among groups in terms of sociodemographic factors and underlying diseases.However,sta-tistically significant differences were observed in the time windows for postoperative pneumonia based on surgery type,incision type,surgical approach and surgery duration(P<0.05).The average time for the onset of postop-erative pneumonia in 263 patients was 2.00(1.00,7.00)days.The postoperative time windows varied for sur-geries involving different systems.The peak incidence occurred on day 0(16 cases)and day 1(17 cases)af-ter neurological surgery,while the peak incidence for digestive system and orthopedic surgeries was on day 1.The time span for the onset of pneumonia after skin surgeries was wider(0-53 days postoperatively)without a clear peak.In addition,33.33%of cardiovascular system surgery cases developed pneumonia 10 days postoperatively.There were also significant time differences in the diagnostic elements of postoperative pneumonia,with fever and abnormal white blood cell counts appearing earlier(median appearance time length:4.00 days)than lung imaging changes(median appearance time length:7.00 days).CONCLUSIONS This study demonstrates significant time differences in the on-set of postoperative pneumonia and confirms the significant spatiotemporal heterogeneity in the diagnostic elements of postoperative pneumonia.These findings provide a quantitative basis for developing dynamic,surgery-type-spe-cific monitoring protocols and prevention and control measures for postoperative pneumonia.
3.Clinical characteristics of postoperative pneumonia patients undergoing different types of surgeries and distribution of onset time
Ran XIN ; Lei QIAO ; Shuangfeng SUN ; Yingying PAN ; Juanjuan KONG ; Hongyu WANG ; Ying YAN
Chinese Journal of Nosocomiology 2025;35(20):3074-3078
OBJECTIVE To explore the time windows for postoperative pneumonia in patients undergoing different surgeries,providing evidence-based references for optimizing infection monitoring and prevention and control strategies.METHODS Sociodemographic characteristics,clinical information and surgical details of 263 patients with postoperative pneumonia from four different types of medical institutions between Jan.2019 and Dec.2024 were retrospectively collected.The time windows for postoperative pneumonia in patients undergoing different surgeries were analyzed.RESULTS There were no statistically significant differences in the time windows for post-operative pneumonia among groups in terms of sociodemographic factors and underlying diseases.However,sta-tistically significant differences were observed in the time windows for postoperative pneumonia based on surgery type,incision type,surgical approach and surgery duration(P<0.05).The average time for the onset of postop-erative pneumonia in 263 patients was 2.00(1.00,7.00)days.The postoperative time windows varied for sur-geries involving different systems.The peak incidence occurred on day 0(16 cases)and day 1(17 cases)af-ter neurological surgery,while the peak incidence for digestive system and orthopedic surgeries was on day 1.The time span for the onset of pneumonia after skin surgeries was wider(0-53 days postoperatively)without a clear peak.In addition,33.33%of cardiovascular system surgery cases developed pneumonia 10 days postoperatively.There were also significant time differences in the diagnostic elements of postoperative pneumonia,with fever and abnormal white blood cell counts appearing earlier(median appearance time length:4.00 days)than lung imaging changes(median appearance time length:7.00 days).CONCLUSIONS This study demonstrates significant time differences in the on-set of postoperative pneumonia and confirms the significant spatiotemporal heterogeneity in the diagnostic elements of postoperative pneumonia.These findings provide a quantitative basis for developing dynamic,surgery-type-spe-cific monitoring protocols and prevention and control measures for postoperative pneumonia.
4. Treatment progress of triple positive breast cancer
Xiaoyu FU ; Deguang KONG ; Juanjuan LI
Chinese Journal of Clinical Pharmacology and Therapeutics 2023;28(8):866-875
Triple positive breast cancer overexpress ER (estrogen receptor), PR and HER2 (human epidermal growth factor receptor 2, HER2), accounting for about 50%-60% of the HER2 positive breast cancer patients. Based on the data from clinical trials, the crosstalk between the ER signaling pathway and the HER2 signaling pathway in triple-positive breast cancer may weaken the efficacy of anti-HER2 therapy and endocrine therapy, and this feature has attracted widespread attention. Emerging evidence shows that while blocking HER2 signaling pathway, together with enhancing blocking of ER signaling pathway, such as anti-HER2 dual-targeting + endocrine therapy ± CDK4 / 6 inhibitors, could effectively overcome drug resistance, and improve the efficacy. Predictive biomarkers including Ki67, intrinsic subtypes, and multi-gene assay, which have the potential benefit for personalized treatment.
5.Efficacy of hyperbaric oxygen on peritoneal dialysis patients with cognitive impairment and its impact on quality of life
Fukun NIU ; Fanling HU ; Shuxia GUO ; Yan ZHUANG ; Xianglei KONG ; Juanjuan SUN ; Fengyu WU
Chinese journal of nautical medicine and hyperbaric medicine 2022;29(5):665-670
Objective:To analyze the effect of hyperbaric oxygen therapy on peritoneal dialysis(PD)patients with cognitive impairment and its impact on quality of life.Methods:A total of 120 maintenance PD patients with cognitive impairment admitted to Linyi Central Hospital from December 2019 to December 2020 were divided into observation group and control group according to the random number table method,with 60 cases in each group. Patients in both groups received conventional treatments including PD for end-stage renal disease. The control group received cognitive training,and the observation group received hyperbaric oxygen therapy on the basis of the training of the control group. The changes of cognitive function and quality of life of the patients in the two groups were compared.Results:There was no significant difference in the levels of BUN,Scr,PaO 2,PaCO 2,and pH between the two groups before and after treatment( P>0.05). After treatment,Hb levels in the two groups were significantly higher than those before treatment,with statistically significant differences( P<0.05),while there was no statistically significant difference between the two groups( P>0.05). After treatment,the Addenbrooke’s Cognitive Examination-Revised(ACE-R)total score and sub-scale scores of patients in both groups were significantly increased than those before treatment,with statistically significant differences( P<0.05),and those scores in the observation group were significantly higher than those in the control group,with statistically significant differences( P<0.05). The serum BNDF level of the observation group was significantly higher than that before treatment. The serum Hcy and CRP levels of the two groups were significantly lower than those before treatment,with statistically significant differences( P<0.05),and those two levels of the observation group were significantly lower than those of the control group,with statistically significant differences( P<0.05). The indicators of quality of life in the two groups were significantly improved than those before treatment,with statistically significant differences( P<0.05),and the indicators of the observation group were better than those of the control group,with statistically significant differences( P<0.05). There was no statistically significant difference in the incidence of adverse events between the two groups during treatment( P>0.05). Conclusion:Hyperbaric oxygen therapy can significantly improve the cognitive function and quality of life of PD patients,and it is safe and reliable,which is worthy of clinical promotion.
6.Efficacy of hyperbaric oxygen on peritoneal dialysis patients with cognitive impairment and its impact on quality of life
Fukun NIU ; Fanling HU ; Shuxia GUO ; Yan ZHUANG ; Xianglei KONG ; Juanjuan SUN ; Fengyu WU
Chinese journal of nautical medicine and hyperbaric medicine 2022;29(5):665-670
Objective:To analyze the effect of hyperbaric oxygen therapy on peritoneal dialysis(PD)patients with cognitive impairment and its impact on quality of life.Methods:A total of 120 maintenance PD patients with cognitive impairment admitted to Linyi Central Hospital from December 2019 to December 2020 were divided into observation group and control group according to the random number table method,with 60 cases in each group. Patients in both groups received conventional treatments including PD for end-stage renal disease. The control group received cognitive training,and the observation group received hyperbaric oxygen therapy on the basis of the training of the control group. The changes of cognitive function and quality of life of the patients in the two groups were compared.Results:There was no significant difference in the levels of BUN,Scr,PaO 2,PaCO 2,and pH between the two groups before and after treatment( P>0.05). After treatment,Hb levels in the two groups were significantly higher than those before treatment,with statistically significant differences( P<0.05),while there was no statistically significant difference between the two groups( P>0.05). After treatment,the Addenbrooke’s Cognitive Examination-Revised(ACE-R)total score and sub-scale scores of patients in both groups were significantly increased than those before treatment,with statistically significant differences( P<0.05),and those scores in the observation group were significantly higher than those in the control group,with statistically significant differences( P<0.05). The serum BNDF level of the observation group was significantly higher than that before treatment. The serum Hcy and CRP levels of the two groups were significantly lower than those before treatment,with statistically significant differences( P<0.05),and those two levels of the observation group were significantly lower than those of the control group,with statistically significant differences( P<0.05). The indicators of quality of life in the two groups were significantly improved than those before treatment,with statistically significant differences( P<0.05),and the indicators of the observation group were better than those of the control group,with statistically significant differences( P<0.05). There was no statistically significant difference in the incidence of adverse events between the two groups during treatment( P>0.05). Conclusion:Hyperbaric oxygen therapy can significantly improve the cognitive function and quality of life of PD patients,and it is safe and reliable,which is worthy of clinical promotion.
7.Screening and analysis of Treponema pallidum specific antibody among childbearing age people in rural areas of Yunnan province, 2013-2017
Zhengyuan XIE ; Guang CAO ; Cai KONG ; Juanjuan CHEN ; Tao WANG ; Shui ZHENG ; Baoxin LI ; Yixiao LI ; Weilei ZU ; Hanfeng YE
Chinese Journal of Epidemiology 2021;42(8):1475-1481
Objective:To determine the prevalence, epidemiological characteristics, and related factors of syphilis infection among rural childbearing age people to promote medical interventions on pre-pregnancy aristogenesis and syphilis infection in Yunnan province.Methods:The subjects in this study were 18-49-year-old rural couples of childbearing age from the National Free Preconception Health Examination Project in Yunnan province during 2013-2017. The descriptive study was carried out to determine the positive rate of Treponema pallidum specific antibody (TPsAb) and related sociodemographic characteristics. Results:The overall positive rate of TPsAb was 0.38% (8 204/2 160 455) in 2 160 455 rural childbearing age people in Yunnan. The positive rate of TPsAb was 0.39% (4 019/1 040 981) in men,higher than that in women (0.37%,4 185/1 119 474). The positive rate of TPsAb was highest in the age group 45-49 years (0.70%,158/22 511). The positive rate of TP antibody in the minority ethnic groups and Han ethnic groups appeared the same (0.38%). However,the highest positive rate of TPsAb was 0.77% (461/60 153) in Hani ethnic group among all the 17 minority ethnic groups. People with education level of primary education had the highest anti-TP positive rate (0.54%,2 327/431 275). The positive rate of TPsAb appeared the highest in Zhaotong (0.73%, 2 049/281 614) area among all the 16 prefectures of the province. The positive rate of TPsAb in the population from the impoverished regions (0.50%,2 963/590 039) was higher than in other disadvantaged areas (0.34%, 3 115/929 368) or areas with average income (0.33%,2 126/641 048). Significant differences appeared in the positive rate of TPsAb among populations of different ages, ethnic, education level, and economic level groups (trend χ2 test, P<0.001). Conclusions:The positive rate of TP antibody in rural childbearing age people in Yunnan was related to multi factors,including sex,age,ethnic group,education level,area, residence, and local economic situation. The positive rate of TPsAb was higher in men than in women. And people with elder age, lower education level, being Hani or Lahu ethnic group, and Zhaotong areas had higher TPsAb positive rates and higher syphilis infection risk.
8.Analysis of HBsAg screening among rural childbearing age people in Yunnan province during 2013-2017
Zhengyuan XIE ; Cai KONG ; Juanjuan CHEN ; Baoxin LI ; Shui ZHENG ; Yixiao LI ; Weilei ZU ; Xin LI ; Yanling YUAN ; Hanfeng YE
Chinese Journal of Epidemiology 2020;41(9):1522-1526
Objective:To explore the current status of HBV infection before pregnancy among rural childbearing age people in Yunnan province, find out the population at high risk and areas with high incidence and provide evidence for promoting pre-pregnancy aristogenesis and medical interventions for prevention of HBV infection.Methods:The subjects in the study were rural childbearing age people aged 18-49 who received the national free pre pregnancy eugenic health examination in Yunnan province during 2013-2017. Descriptive analysis was conducted for HBsAg positive rate and its difference among various characteristics. The SPSS 22.0 software was used for statistical analysis.Results:A total of 2 180 841 rural childbearing age people were surveyed, the HBsAg positive rate before pregnancy was 3.13%(68 314/2 180 841) in Yunnan. And the HBsAg positive rate was 3.56% (37 694/1 057 376) in men, higher than that in women (2.73%, 30 620/1 123 465). The HBsAg positive rate was highest in age group 41-49 years (3.77%, 3 126/82 931). The HBsAg positive rate was higher in minority ethnic group (3.77%, 27 974/742 518) than in Han ethnicity group (2.81%, 40 024/1 425 870). Moreover, the highest HBsAg positive rate of was 8.20%(851/10 378) in Yao ethnic group. The people with education level of primary education had the highest HBsAg positive rate (3.97%, 17 294/435 219). The HBsAg positive rate was highest in Xishuangbanna Dai autonomous prefecture (6.65%, 3 875/58 232) in Yunnan. Moreover, the HBsAg positive population in severe poor area (3.32%, 19 755/595 665) was higher than those in less poor area (3.17%, 29 750/939 101) or normal areas (2.91%, 18 809/646 075). There was significant difference in HBsAg positive rate among populations in different age groups, ethnic groups, education level groups, areas and economic level groups ( P<0.001). Conclusions:The HBsAg positive rate in rural childbearing age people in Yunnan was related to multi factors, including sex, age, ethnic group, education level, area and economic development level. The HBsAg positive rate was higher in men, older age and lower education level, minority ethnic group (especially in Yao ethnic group), from Xishuangbanna prefecture and the poor living conditions in childbearing age population.
10.The characteristics of the surface electromyographic signals recorded from submental and infrahyoid muscles when swallowing in patients with nasopharyngeal carcinoma after radiotherapy
Juanjuan HE ; Xiaomei WEI ; Qingcong KONG ; Chunqing XIE ; Zulin DOU
Chinese Journal of Physical Medicine and Rehabilitation 2017;39(12):899-902
Objective To study the surface electromyographic signal characteristics of submental and infrahyoid muscles when swallowing in patients with nasopharyngeal carcinoma (NPC) following radiotherapy.Methods Fifteen post-radiotherapy NPC patients with pharyngeal dysphagia confirmed by videofluoroscopie swallowing study (VFSS) were selected into the patient group,while another 15 healthy subjects were recruited into the healthy group.Surface electomyography (sEMG) was used to collect sEMG signals from submental muscles and infrahyoid muscles during both dry and 3 ml thick liquid swallowing,and sEMG duration,average amplitude and peak amplitude parameters were compared between the two groups.Results Compared with healthy subjects,the patient group showed significantly longer sEMG duration of submental muscles and infrahyoid muscles during dry swallowing and 3 ml thick liquid swallowing (P<0.05).Moreover,significantly longer sEMG duration of submental muscles and infrahyoid muscles was observed when swallowing 3 ml thick liquid,compared with dry swallowing (P<0.05).The average sEMG amplitude and peak amplitude of submental muscles and infrahyoid muscles of patients when dry swallowing and 3 ml thick liquid swallowing were both significantly higher than those of healthy subjects (P<0.05).Conclusion Submental muscles and infrahyoid muscles manifested enhanced contractions during swallowing in NPC patients with dysphagia,which played a compensatory role after radiotherapy.

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