1.Establishment and verification of ultra high performance liquid chromatography and tandem mass spectrometry for determination of receptor binding domain protein content of virus-like particle SARS-CoV-2 vaccine
Chinese Journal of Biologicals 2025;38(10):1211-1218+1225
Objective To establish an ultra high performance liquid chromatography and tandem mass spectrometry(UHPLCMS/MS) method for determination of the protein content of the receptor binding domain(RBD) of virus-like particle(VLP)SARS-CoV-2 vaccine, and to validate and preliminarily apply it, so as to provide technical support for the quality control.Methods The characteristic peptide of RBD protein of SARS-CoV-2 vaccine was screened by high resolution mass spectrometer, and the content of RBD protein was determined by UHPLC-MS/MS. The separation was performed on a Thermo Fisher Hypersil GOLD C18(100 mm × 2. 1 mm, 3 μm) using the mobile phase A consisting of 0. 1% formic acid aqueous solution and mobile phase B consisting of 0. 1% acetonitrile formate solution at a flow rate of 0. 3 mL/min, via the gradient elution,0-3 min,2% B;3-10 min,2%→50% B;10-10.1 min,50%→90% B;10.1-12 min,90% B;12-12.1 min,90%→2% B; 12. 1-15 min, 2% B. The column temperature was 40 ℃ and the sample size was 1 μL. The specificity, stability,precision, linearity, limit of quantitation(LOQ), recovery and repeatability of the method were verified. Six batches of VLP SARS-CoV-2 vaccine bulk solution and six batches of finished products were detected by the method, and the correlation between the method and the method by amino acid analyzer was analyzed.Results The blank solution had no interference with the determination of characteristic peptide content. The test product solution showed good stability within 24 h at room temperature. The relative standard deviation(RSD) of the peak area of the characteristic peptide was 1. 0% after six consecutive injections of the characteristic peptide reference solution. The standard curve had a good linear relationship at the characteristic peptide concentration ranged from 0. 008 954 to 0. 358 200 μg/mL, and the LOQ was 0. 008 954 μg/mL. The RSDs of recovery rates of characteristic peptide in bulk solution and vaccine samples were all less than 10%. The contents of six batches of vaccine bulk solution and six batches of vaccine samples were 0. 169 40-0. 196 60 mg/mL and 0. 014 35-0. 020 50 mg/mL, respectively. The deviation of RBD protein content in VLP SARS-CoV-2 vaccine bulk solution determined by amino acid analyzer and characteristic peptide method was 5. 3%, and the determination results were basically the same.Conclusion The established UHPLC-MS/MS method is simple to operate with strong specificity and accurate quantitation,and can be used for the determination of RBD protein content in VLP SARS-CoV-2 vaccine.
2.Self-sufficient nanoparticles with dual-enzyme activity trigger radical storms and activate cascade-amplified antitumor immunologic responses.
Liping BAI ; Jin YANG ; Siting YU ; Zhongzheng XIANG ; Yuanyuan ZENG ; Meiling SHEN ; Xiaorong KOU ; Qinjie WU ; Changyang GONG
Acta Pharmaceutica Sinica B 2024;14(2):821-835
Radiotherapy (RT) can potentially induce systemic immune responses by initiating immunogenic cell death (ICD) of tumor cells. However, RT-induced antitumor immunologic responses are sporadic and insufficient against cancer metastases. Herein, we construct multifunctional self-sufficient nanoparticles (MARS) with dual-enzyme activity (GOx and peroxidase-like) to trigger radical storms and activate the cascade-amplified systemic immune responses to suppress both local tumors and metastatic relapse. In addition to limiting the Warburg effect to actualize starvation therapy, MARS catalyzes glucose to produce hydrogen peroxide (H2O2), which is then used in the Cu+-mediated Fenton-like reaction and RT sensitization. RT and chemodynamic therapy produce reactive oxygen species in the form of radical storms, which have a robust ICD impact on mobilizing the immune system. Thus, when MARS is combined with RT, potent systemic antitumor immunity can be generated by activating antigen-presenting cells, promoting dendritic cells maturation, increasing the infiltration of cytotoxic T lymphocytes, and reprogramming the immunosuppressive tumor microenvironment. Furthermore, the synergistic therapy of RT and MARS effectively suppresses local tumor growth, increases mouse longevity, and results in a 90% reduction in lung metastasis and postoperative recurrence. Overall, we provide a viable approach to treating cancer by inducing radical storms and activating cascade-amplified systemic immunity.
3.Application of Party building-guided quality control circle management in emergency services
Chengquan ZHAI ; Liping WANG ; Shoujing GONG ; Yadong LI ; Jilai QU ; Yugui HAO
Modern Hospital 2024;24(8):1175-1177,1181
Objective To investigate the role of quality control circle management guided by Party building philosophy in improving patient satisfaction in emergency services.Methods Quality control circle management guided by the Party building philosophy was implemented in May 2023.Two groups of 1 000 emergency patients each were selected,one from April to May 2023(pre-implementation)and the other from April to May 2024(post-implementation).The control group and the observation group were compared across several metrics in terms of satisfaction scores,satisfaction rates,120 emergency call response time,3-minute pre-hospital emergency dispatch rates,and average waiting times for emergency observation patients.Results The ob-servation group demonstrated higher satisfaction scores,satisfaction rates,and higher 3-minute pre-hospital emergency dispatch rates compared to the control group.Additionally,the observation group had shorter emergency call response time and average waiting time in the emergency department,with statistically significant differences(P<0.05).Conclusion The integration of quality control circle management guided by Party construction philosophy in the management of emergency patients has been shown to address operational issues,optimize service processes,and significantly improve patient satisfaction and experience.This method is worthy of broader application and promotion in clinical practice.
4.Mechanism of Action of Coptidis Rhizoma and Ophiopogonis Radix in Delaying Diabetic Nephropathy Based on EGFR/PI3K/Akt Signaling Pathway
Shaoyu LI ; Man GONG ; Qiufang LI ; Liping DAI ; Guiqun WANG ; Qiuchen YANG ; Qiongqiong ZHANG ; Erping XU ; Yalin LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(20):22-29
ObjectiveTo observe the effect of Coptidis Rhizoma and Ophiopogonis Radix on renal tissue injury and epidermal growth factor receptor (EGFR)/phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway in rats with diabetic nephropathy (DN) and explore its possible mechanism of delaying DN. MethodThirty-six male Wistar rats were randomly divided into a normal group (6 rats) and a model group (30 rats). The model group was fed with a high-fat and high-sugar diet combined with streptozotocin (STZ) to establish a rat model of type 2 diabetes. After the successful preparation of the model, the rats were randomly divided into the model group, low, medium, and high dose groups of Coptidis Rhizoma and Ophiopogonis Radix (100, 200, 400 mg·kg-1), and metformin group (200 mg·kg-1). After administration, the levels of fasting blood glucose (FBG), 24 h urine protein (24 h-UTP), creatinine (SCr), urea nitrogen (BUN), and uric acid (UA) were detected. Hematoxylin-eosin (HE) staining and Masson staining were used to observe the pathological changes of renal tissue in rats. Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were used to detect the related protein expression of EGFR, PI3K, and Akt and their mRNA expression levels in the renal tissue of rats in each group. ResultsCompared with the normal group, the levels of FBG, SCr, BUN, UA, 24 h-UTP, and kidney index in the model group were significantly increased (P<0.01), most renal tubular epithelial cells were necrotic, and the content of collagen in glomeruli was significantly increased (P<0.01). Compared with the model group, the above indexes of rats in each administration group were improved to varying degrees. The FBG, SCr, BUN, UA, 24 h-UTP, and kidney index of rats in each dose group and metformin group were significantly decreased (P<0.01, P<0.05). The necrosis degree of renal tubular epithelial cells was reduced, and the fibrosis area was decreased (P<0.01). There related protein and mRNA expressions of EGFR, PI3K, and Akt were significantly increased (P<0.05, P<0.01). ConclusionCoptidis Rhizoma and Ophiopogonis Radix can alleviate renal tissue injury in rats with DN, and their mechanism may be related to the regulation of the EGFR/PI3K/Akt signaling pathway.
5.Advances in Immunological Studies of Targeted Radionuclide Therapy
Liping YANG ; Yuxuan ZHENG ; Zhuoling RAN ; Rui HU ; Yuwei LIANG ; Yue SUN ; Chang LU ; Huijie LI ; Xuhan LI ; Jian GONG
Herald of Medicine 2024;43(10):1598-1602
The killing effect of radiation therapy on healthy cells has led to the creation of targeted radionuclide therapy,which effectively reduces the damage to surrounding normal cells.At present,alpha(α)and beta(β)radionuclides are the research hotspots of targeted therapy.Numerous preclinical and clinical studies have shown that radiation therapy not only has local anti-tumor effects,but also exerts systemic anti-tumor effects by triggering the body's immune response.This paper describes in detail the characteristics and clinical applications of commonly used radionuclides,and discusses the mechanism of radiation-triggered body immune response as well as the related research on the combined use of radiation therapy,targeted radionuclide therapy and immunotherapy.
6.Application and Prospect of Therapeutic Radionuclides
Hongzhu LIU ; Liping YANG ; Yuxuan ZHENG ; Chang LU ; Ran ZHANG ; Yuwei LIANG ; Xu GAO ; Zhenjiang ZHANG ; Minghui AN ; Jing XIE ; Jian GONG
Herald of Medicine 2024;43(10):1603-1609
Targeted radiation therapy using radionuclides is a favored approach for treating tumors.This procedure involves the delivery of drugs to the lesion site via carriers or interventional methods,followed by the emission of radiation energy that selectively irradiates the lesion tissue.This approach minimizes damage to normal tissue and achieves the desired therapeutic effect.Factors such as the type of therapeutic radionuclide,radiation energy,physical half-life,method of preparation,and toxicity determine their clinical application.In this paper,the characteristics and clinical application of therapeutic radionuclides were reviewed to providing reference for the clinical application of targeted therapeutic radionuclides.
7.Applications and Prospect of Diagnostic Radionuclide
Jiaxin DING ; Zhuoling RAN ; Yuxian ZHANG ; Ran ZHANG ; Lin YU ; Liping YANG ; Yuanqing NING ; Xu GAO ; Minghui AN ; Jing XIE ; Dong CHAI ; Jian GONG
Herald of Medicine 2024;43(10):1609-1614
Nuclear medicine plays an indispensable role in the diagnosis,treatment,and prognosis of a wide range of diseases.Nuclear medicine using radionuclides for diagnosis has the advantages of accuracy,speed,high sensitivity and high resolution.Currently,several radionuclides play pivotal roles in disease diagnosis.This article primarily examines the clinical application and research of diagnostic radionuclides,including 18 F,89 Zr,68 Ga,99m Tc,131 I,123 I,and 11 C.The objective is to offer valuable insights for disease diagnosis and staging of diseases.
8.The role of NLRP3 signaling pathway in allergic rhinoconjunctivitis
Yubo GONG ; Xiaohua GUO ; Wen-Jun LU ; Yuanchao LI ; Changyu QIU ; Yuanyuan SHI ; Liping XIA ; Lin SHI ; Wei WU ; Ling LUO
The Journal of Practical Medicine 2024;40(14):1922-1927
Objective The objective of this study was to establish a mouse model of allergic rhinoconjunctivitis and investigate the role of the NLRP3 signaling pathway in allergic rhinoconjunctivitis.Methods Thirty-three female C57 mice(SPF)were randomLy divided into 3 groups:the control group,the experimental group,and the NLRP3-/-group.On days 0,4,7,14,and 21,the experimental group and NLRP3-/-group received a 0.2 mL intraperitoneal injection of medicine containing OVA(100 μg)and adjuvant Al(OH)3(4 mg),respectively.After an interval of 3 days,each eye and nose were dosed with 10 μL of 5%OVA for five consecutive days a week to induce allergic symptoms.During sensitization and excitation stages,the control group was replaced with an equiva-lent amount of PBS.Ocular and nasal symptoms were observed and scored.The levels of OVA-specific IgE,IL-4,IL-17,and IL-18 in serum were measured using ELISA,while changes in palpebral conjunctiva and nasal mucosa were assessed by hematoxylin-eosin staining.The expression of NLRP3 mRNA in conjunctival tissue and nasal mucosa was determined using real-time PCR analysis.Statistical analysis was performed using SPSS17.0 software with P<0.05 considered as statistically significant difference.Results The experimental group and NLRP3-/-group exhibited induced nasal and ocular allergic symptoms.In the experimental group,the duration of nasal allergy symptoms was(10.500±1.080)days,while the duration of eye allergy symptoms was(20.300±2.058)days.In the NLRP3-/-group,the duration of nasal allergy symptoms was(13.400±1.955)days,and for eye allergy symp-toms it was(20.900±2.132)days.The duration of nasal allergies in the NLRP3-/-group significantly exceeded that in the experimental group(P<0.05),whereas there were no significant differences observed in eye allergy durations between these two groups(P>0.05).Levels of OVA-specific IgE,IL-4,and IL-17 were significantly higher in both the experimental and NLRP3-/-groups compared to those in the control group(P<0.05).Additionally,serum IL-18 content increased significantly in the experimental group when compared with both control and NLRP3-/-groups(P<0.05).Conjunctival tissue lesions as well as nasal mucosa damage were evident in both experimental and NLRP3-/-groups.mRNA expression levels of NLRP3 within conjunctival tissue and nasal mucosa from the experimental group showed a significant increase when compared to those from both control and NLRP3-/-groups(P<0.05).Conclusion Allergic rhinoconjunctivitis pathogenesis is influenced by various factors;however,the involvement of NLPR3 signaling pathway promotes its development.
9.Associations of health literacy with metabolic status and quality of life among comunity patients with type 2 diabetes in Shanghai
Liping YANG ; Haihong SONG ; Jue SHEN ; Jinli GAO ; Zhiping ZHANG ; Liuyu QIAN ; Huan GONG
Shanghai Journal of Preventive Medicine 2024;36(11):1051-1059
ObjectiveTo investigate the current status of health literacy among type 2 diabetes mellitus patients in Shanghai communities, to analyze its correlation with glycemic and lipid metabolic indicators and quality of life (QoL), and to provide an evidence for the development of effective intervention measures. MethodsA follow⁃up survey was conducted among diabetic patients diagnosed at hospitals above the secondary level from March to June, 2023, who were part of a diabetes cohort study jointly established by Songnan Community in Baoshan District and the Shanghai Institute of Endocrinology. Patients were assessed using a simplified evaluation form for diabetes-related health literacy and a QoL scale, and metabolic indicators such as blood glucose and blood lipids were measured. Pearson correlation coefficients were employed to assess the relationship between health literacy levels and metabolic indicators as well as QoL. Logistic regression models were used to estimate the odds ratios (OR) and 95% confidence intervals (CI) for the associations between patients’ health literacy levels, metabolic status, and QoL. ResultsAmong the 932 diabetic patients surveyed, the health literacy level was low, with an average score of (65.2±13.4) points of which (62.6±13.8) points for males and (67.9±12.5) points for females. The average QoL score was (44.1±9.2) points of which (44.0±9.5) points for males and (44.2±8.9) points for females. The achievement rates of HbA1c, total cholesterol (TC), and triglycerides (TG) were 51.6%, 30.3%, and 64.8%, respectively. The TC achievement rate among male patients (37.6%) was significantly higher than that among females (22.8%). Significant differences in HbA1c achievement rates were observed across age groups, education levels, disease durations, and BMI levels (P<0.05). Similarly, significant differences were found in FPG achievement rates across different age groups, annual household incomes, disease durations, and family histories (P<0.05). No statistically significant differences were noted in TG and HDL-C achievement rates across all variables (P>0.05). However, LDL-C achievement rates were significantly higher in patients without complications compared to those with complications (P<0.05). Health literacy showed a significant negative correlation with psychological dimension, treatment dimension, and QoL scores (r=-0.14, P<0.05). After adjusting for potential confounding factors, a significant association was found between patients’ health literacy levels and QoL, but not with metabolic indicators. ConclusionThere is a significant correlation between health literacy levels and QoL among diabetic patients in community settings. Further exploration is needed to identify factors influencing patients’ QoL and glycemic and lipid metabolic indicators.
10.Latent tuberculosis infection status and its risk factors among tuberculosis-related health-care workers in Shanghai
Lixin RAO ; Wei SHA ; Huili GONG ; Lihong TANG ; Liping LU ; Yan LIU ; Zheyuan WU ; Zurong ZHANG ; Xin SHEN ; Qingwu JIANG
Shanghai Journal of Preventive Medicine 2023;35(3):203-207
ObjectiveTo obtain the status of latent tuberculosis infection (LTBI) among tuberculosis (TB)-related health-care workers (HCWs) in Shanghai, and to explore the risk factors related to TB infection. MethodsA multi-center cross-sectional study was conducted by recruiting medical workers from multiple designated TB hospitals, centers for disease control and prevention, and community health service centers in Shanghai. Each subject was required to complete a questionnaire and to provide a blood sample for TB infection test. Univariate and multivariate analysis ware made in order to find risk factors relating to TB infection. ResultsA total of 165 medical workers were recruited, and the proportion of TB infection was 16.36% (95%CI: 11.49%‒22.76%). Multivariate logistic analysis showed that clinical doctors and nurses (adjusted OR=9.756, 95%CI: 1.790‒53.188), laboratory staffs (adjusted OR=78.975, 95%CI: 8.749‒712.918), and nursing and cleaning workers (adjusted OR=89.920, 95%CI: 3.111‒2 598.930) had higher risk of TB infection. ConclusionThe overall LTBI prevalence among TB-related HCWs is low. However, working as doctors, nurses, laboratory staffs, nursing workers and cleaning workers are risk factors of TB infection. TB-related HCWs who work at hospitals are at risk of TB infection comparing to medical staffs who work outside hospitals.


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