1.Preparation and identification of monoclonal antibodies against cat allergen Fel d 1.
Linying CAI ; Zichen ZHANG ; Zhuangli BI ; Shiqiang ZHU ; Miao ZHANG ; Yiming FAN ; Jingjie TANG ; Aoxing TANG ; Huiwen LIU ; Yingying DING ; Chen LI ; Yingqi ZHU ; Guijun WANG ; Guangqing LIU
Chinese Journal of Cellular and Molecular Immunology 2025;41(4):348-354
Objective Currently, there is no commercially available quantitative detection kit for the main Felis domestic allergen (Fel d 1) in China. To establish a rapid detection method for Fel d 1, this study aims to prepare monoclonal antibodies against Fel d 1 protein. Methods The codon preference of Escherichia coli was utilized to optimize and synthesize the Fel d 1 gene. The prokaryotic expression plasmid pET-28a-Fel d 1 was constructed and used to express and purify the recombinant Fel d 1 protein. Subsequently, the recombinant protein was immunized into BALB/c mice and monoclonal antibodies (mAbs) were prepared by the hybridoma technique. An indirect ELISA was established using the recombinant Fel d 1 as the coating antigen, and hybridoma cell lines were screened for positive clones. The specificity and antigenic epitopes of the mAbs were confirmed by Western blot analysis. Finally, the selected hybridoma cells were injected into the peritoneal cavities of BALB/c mice for large-scale monoclonal antibody production. Results The recombinant plasmid pET-28a-Fel d 1 was successfully constructed, and soluble Fel d 1 protein was obtained after optimizing the expression conditions. Western blot and antibody titer assays confirmed the successful isolation of two hybridoma cell lines, 7D11 and 5H4, which stably secreted mAbs specific to Fel d 1. Antibody characterization revealed that the 5H4 mAb was of the IgG2a subtype and could recognize the amino acid region 105-163 of Fel d 1, while the 7D11 mAb was the IgG1 subtype and could recognize the amino acid region 1-59. Conclusion The high-purity recombinant Fel d 1 protein produced in this study provides a promising alternative for clinical immunotherapy of cat allergies. Furthermore, the monoclonal antibody prepared in this experiment lays a material foundation for the in-depth study of the biological function of Fel d 1 and the development of ELISA detection.
Animals
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Antibodies, Monoclonal/biosynthesis*
;
Mice, Inbred BALB C
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Cats
;
Mice
;
Allergens/genetics*
;
Glycoproteins/genetics*
;
Enzyme-Linked Immunosorbent Assay
;
Hybridomas/immunology*
;
Recombinant Proteins/genetics*
;
Female
;
Antibody Specificity
2.SnoRNAs: The promising targets for anti-tumor therapy.
Xiaoyun HU ; Wanlin CUI ; Min LIU ; Fangxiao ZHANG ; Yingqi ZHAO ; Mingrong ZHANG ; Yuhang YIN ; Yalun LI ; Ying CHE ; Xianglong ZHU ; Yuxuan FAN ; Xiaolan DENG ; Minjie WEI ; Huizhe WU
Journal of Pharmaceutical Analysis 2024;14(11):101064-101064
Recently, small nucleolar RNAs (snoRNAs) have transcended the genomic "noise" to emerge as pivotal molecular markers due to their essential roles in tumor progression. Substantial evidence indicates a strong association between snoRNAs and critical clinical features such as tumor pathology and drug resistance. Historically, snoRNA research has concentrated on two classical mechanisms: 2'-O-ribose methylation and pseudouridylation. This review specifically summarizes the novel regulatory mechanisms and functional patterns of snoRNAs in tumors, encompassing transcriptional, post-transcriptional, and post-translational regulation. We further discuss the synergistic effect between snoRNA host genes (SNHGs) and snoRNAs in tumor progression. More importantly, snoRNAs extensively contribute to the development of tumor cell resistance as oncogenes or tumor suppressor genes. Accordingly, we provide a comprehensive review of the clinical diagnosis and treatment associated with snoRNAs and explore their significant potential as novel drug targets.
3.Vitamin D receptor (VDR) mediates the quiescence of activated hepatic stellate cells (aHSCs) by regulating M2 macrophage exosomal smooth muscle cell-associated protein 5 (SMAP-5).
Xuwentai LIU ; Yue WU ; Yanyi LI ; Kaiming LI ; Siyuan HOU ; Ming DING ; Jingmin TAN ; Zijing ZHU ; Yingqi TANG ; Yuming LIU ; Qianhui SUN ; Cong WANG ; Can ZHANG
Journal of Zhejiang University. Science. B 2023;24(3):248-261
An effective therapeutic regimen for hepatic fibrosis requires a deep understanding of the pathogenesis mechanism. Hepatic fibrosis is characterized by activated hepatic stellate cells (aHSCs) with an excessive production of extracellular matrix. Although promoted activation of HSCs by M2 macrophages has been demonstrated, the molecular mechanism involved remains ambiguous. Herein, we propose that the vitamin D receptor (VDR) involved in macrophage polarization may regulate the communication between macrophages and HSCs by changing the functions of exosomes. We confirm that activating the VDR can inhibit the effect of M2 macrophages on HSC activation. The exosomes derived from M2 macrophages can promote HSC activation, while stimulating VDR alters the protein profiles and reverses their roles in M2 macrophage exosomes. Smooth muscle cell-associated protein 5 (SMAP-5) was found to be the key effector protein in promoting HSC activation by regulating autophagy flux. Building on these results, we show that a combined treatment of a VDR agonist and a macrophage-targeted exosomal secretion inhibitor achieves an excellent anti-hepatic fibrosis effect. In this study, we aim to elucidate the association between VDR and macrophages in HSC activation. The results contribute to our understanding of the pathogenesis mechanism of hepatic fibrosis, and provide potential therapeutic targets for its treatment.
Humans
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Hepatic Stellate Cells/pathology*
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Receptors, Calcitriol
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Liver Cirrhosis/pathology*
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Macrophages/metabolism*
4.Rapid health technology assessment of 4 kinds of drugs for membranous nephropathy
Shengyu ZHANG ; Laixinyue SHU ; Pengli ZHU ; Lijuan NING ; Yongwu CHEN ; Fei WU ; Yingqi WU ; Aizong SHEN
China Pharmacy 2023;34(8):988-992
OBJECTIVE To compare the efficacy, safety and economy of tacrolimus (TAC), cyclosporin A (CsA), cyclophosphamide (CTX) and rituximab (RTX) in the treatment of membranous nephropathy (MN). METHODS Retrieved from Pubmed, the Cochrane Library, Wanfang data, CNKI and health technology assessment (HTA) official website, HTA reports, systematic reviews/meta-analysis and pharmacoeconomic studies about TAC, CsA, CTX and RTX combined with glucocorticoid in the treatment of MN were collected during the inception and Mar. 2022. After data extraction and quality evaluation, descriptive analysis was performed on the results of the included studies. RESULTS A total of 15 articles were included, involving 13 systematic reviews/meta-analysis and 2 pharmacoeconomic studies. In terms of efficacy, TAC and CsA showed significant advantages in increasing the response rate, and could improve the levels of urine protein, serum albumin, serum creatinine and serum total cholesterol. In terms of safety, the incidence of adverse reaction induced by TAC, CsA and RTX was low and the symptoms were mild. In terms of economics, CTX cost lower but caused severe adverse reaction; TAC cost higher but showed higher remission rate and good safety. CONCLUSIONS TAC combined with glucocorticoid may be the recommended scheme for MN.
5.Efficacy of human umbilical cord mesenchymal stem cells via different transplantation approaches in treatment of rats with liver cirrhosis
Yingqi QIU ; Hongwei WANG ; Hongyan ZHU ; Hongliang YU ; Fan XIE ; Cuibao JIANG
Journal of Clinical Hepatology 2023;39(12):2851-2857
ObjectiveTo investigate the therapeutic effect of the frozen and fresh preparations of human umbilical cord mesenchymal stem cells (hUC-MSC) on a rat model of liver cirrhosis after transplantation via the portal vein or the caudal vein. MethodsA total of 70 specific pathogen-free healthy male Sprague-Dawley rats were randomly divided into normal group (13 rats fed with ordinary tap water and rat food) and liver cirrhosis model group (57 rats given subcutaneous multi-point injection of mixed carbon tetrachloride/olive oil solution). At week 8, the growth of rats was observed for both groups, and 3 rats were selected from each group for histopathological examination to confirm the formation of liver cirrhosis. A total of 50 rats were selected from the liver cirrhosis model and were divided into model group, portal vein group+fresh cell preparation group, portal vein+frozen cell preparation group, caudal vein+fresh cell preparation group, and caudal vein+frozen cell preparation group using a random number table, with 10 rats in each group. Fresh or frozen hUC-MSC were transplanted via the portal vein or the caudal vein, and after 4 weeks of administration, the different groups were compared in terms of the changes in liver function parameters and liver fibrosis degree. Continuous data were expressed as mean±standard deviation, and the independent-samples t test was used for comparison between two groups; a one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsAt week 8 of modeling, the model group showed the formation of pseudolobules of different sizes in the liver and met the diagnostic criteria for liver cirrhosis, with significant increases in the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBil), and alkaline phosphatase (ALP) compared with the normal group (all P<0.001), suggesting that the rat model of liver cirrhosis was established successfully. There were significant differences in the levels of ALT, AST, TBil, and ALP between the five groups (F=232.00, 177.10, 112.30, 121.70, all P<0.001). Further comparison between two groups showed that the model group had significantly higher levels of ALT, AST, TBil, and ALP than the normal group (all P<0.01), and the portal vein group+fresh cell preparation group, the portal vein+frozen cell preparation group, the caudal vein+fresh cell preparation group, and the caudal vein+frozen cell preparation group had significantly lower levels of ALT, AST, TBil, and ALP than the model group (all P<0.01). ConclusionThere are significant improvements in liver function and liver fibrosis degree in a rat model of liver cirrhosis at week 4 after the transplantation of hUC-MSC, and frozen or fresh cell preparation and different transplantation approaches have no significant influence on treatment outcome.
6.Single-cell RNA sequencing reveals the dynamics of hepatic non-parenchymal cells in autoprotection against acetaminophen-induced hepatotoxicity
Lingqi YU ; Jun YAN ; Yingqi ZHAN ; Anyao LI ; Lidan ZHU ; Jingyang QIAN ; Fanfan ZHOU ; Xiaoyan LU ; Xiaohui FAN
Journal of Pharmaceutical Analysis 2023;13(8):926-941
Gaining a better understanding of autoprotection against drug-induced liver injury(DILI)may provide new strategies for its prevention and therapy.However,little is known about the underlying mechanisms of this phenomenon.We used single-cell RNA sequencing to characterize the dynamics and functions of hepatic non-parenchymal cells(NPCs)in autoprotection against DILI,using acetaminophen(APAP)as a model drug.Autoprotection was modeled through pretreatment with a mildly hepatotoxic dose of APAP in mice,followed by a higher dose in a secondary challenge.NPC subsets and dynamic changes were identified in the APAP(hepatotoxicity-sensitive)and APAP-resistant(hepatotoxicity-resistant)groups.A chemokine(C-C motif)ligand 2+endothelial cell subset almost disappeared in the APAP-resistant group,and an R-spondin 3+endothelial cell subset promoted hepatocyte proliferation and played an important role in APAP autoprotection.Moreover,the dendritic cell subset DC-3 may protect the liver from APAP hepatotoxicity by inducing low reactivity and suppressing the autoimmune response and occurrence of inflammation.DC-3 cells also promoted angiogenesis through crosstalk with endothelial cells via vascular endothelial growth factor-associated ligand-receptor pairs and facilitated liver tissue repair in the APAP-resistant group.In addition,the natural killer cell subsets NK-3 and NK-4 and the Sca-1-CD62L+natural killer T cell subset may promote autoprotection through interferon-y-dependent pathways.Furthermore,macrophage and neutrophil subpopulations with anti-inflammatory phenotypes promoted tolerance to APAP hepatotoxicity.Overall,this study reveals the dynamics of NPCs in the resistance to APAP hepatotoxicity and provides novel insights into the mechanism of autoprotection against DILI at a high resolution.
7.Construction of post competency evaluation index system for Da Vinci robot specialist nurses based on onion model
Chenjie YANG ; Yingqi CHEN ; Ning ZHOU ; Tingting FU ; Hui FEI ; Hong ZHU ; Xinger QIAN
Chinese Journal of Modern Nursing 2023;29(33):4494-4500
Objective:To construct the post competency evaluation index system for Da Vinci robot specialist nurses based on onion model, so as to provide references for the training and evaluation of Da Vinci robot specialist nurses in China.Methods:From August to September 2022, using the "onion model" as the theoretical framework, a preliminary draft of the index system was developed using domestic and foreign literature review, semi-structured interviews and group discussions. The Delphi method was applied to 22 nursing experts in the relevant fields to conduct two rounds of expert correspondence, screen and modify the items, and calculate the weight using the analytic hierarchy process. Finally, the post competency index system for Da Vinci robot specialist nurses based on the onion model was formed.Results:Two rounds of expert correspondence were conducted. The positive coefficient of experts was 90.91% (20/22) in the first round and 100.00% (20/20) in the second round, the coefficient of expert authority was 0.840 and 0.873, and the Kendall's coordination coefficient was 0.181 and 0.324 ( P<0.01), respectively. The final post competency evaluation index system for Da Vinci robot specialist nurses consisted of 4 first-level indicators, 14 second-level indicators and 58 third-level indicators. Conclusions:This study constructs a comprehensive post competency evaluation index system for Da Vinci robot specialist nurses based on onion model, with high expert enthusiasm and authority. It can provide references for the training and evaluation of Da Vinci robot specialist nurses in China.
8.Efficacy and safety of albumin-bound paclitaxel in the treatment of advanced non-small cell lung cancer
Peng JI ; Lijuan NING ; Yongwu CHEN ; Pengli ZHU ; Fei WU ; Yingqi WU ; Hui YAN ; Yadi GENG ; Shengyu ZHANG ; Aizong SHEN
China Pharmacy 2022;33(22):2762-2765
OBJECTIVE To observe the efficacy and safety of albumin-bound paclitaxel in the treatment of advanced non- small cell lung cancer (NSCLC). METHODS Clinical data of patients with advanced NSCLC treated in our hospital from January 2018 to December 2021 were selected. According to their chemotherapy regimen,they were divided into albumin-bound paclitaxel group and paclitaxel group, with 100 patients in each group. Both groups received chemotherapy regimen containing Paclitaxel for injection (albumin-bound) or Paclitaxel injection for at least 2 cycles (every 21 days as a cycle). The progression-free survival (PFS) and efficacy of the two groups were compared,and the occurrence of toxic and side effects were recorded. RESULTS The patients in albumin-bound paclitaxel group completed 430 cycles of chemotherapy, with an average of 4.3 cycles; patients in paclitaxel group completed 476 cycles of chemotherapy, with an average of 4.8 cycles. The median PFS (4.0 months) and the response rate (13.00%) of albumin-bound paclitaxel group were not significantly different from those of paclitaxel group (4.0 months,9.00%) (P>0.05). The disease control rate (99.00%) was significantly higher than that in paclitaxel group (89.00%), and the incidences of leukopenia, neutropenia, thrombocytopenia,anemia, sensory neuropathy, fatigue,nausea and vomiting,joint myalgia in albumin-bound paclitaxel group were significantly lower than those in paclitaxel group (P<0.05). CONCLUSIONS Albumin-bound paclitaxel is effective in the treatment of advanced NSCLC, and it can better control the progression of the disease and is safer than ordinary paclitaxel.
9.Disinfection quality of secondary and primary schools and kindergartens in Wuxi
ZOU Yaming, LIU Yunhua, WANG Hesheng, YOU Yingqi, LAN Cejie, ZHU Ding
Chinese Journal of School Health 2021;42(2):291-293
Objective:
To comprehensively understand the disinfection quality of secondary and primary schools and kindergartens in Wuxi, so as to find out problems and to provide advice for improvement.
Methods:
Stratified random sampling method was applied in the investigation to select 73 schools and classes. The qualities of room air, surface of object, hand hygiene of staff, tableware, ultraviolet lamp and disinfectant in use (including bacterial contamination and concentration of chlorine-containing) were all tested.
Results:
A total of 2 563 samples were collected with the total disinfection qualified rate 88.02%. The qualified rates of kindergarten, primary and secondary schools were 87.89%, 90.67% and 85.83% respectively. The rates of Xishan and Jiangyin districts were 75.24% and 75.89% respectively. The quality of urban schools was better than the rural(χ 2=16.57, P<0.01). There was no statistical significance between public and private schools (χ 2=0.01, P=0.92). The rank of qualified rates of different objects was: bacterial contamination (100.00%) > room air (93.13%) > tableware (91.87%) > surface of object (89.40%) > ultraviolet lamp (84.00%) > concentration of chlorine-containing (73.68%) > hand hygiene of staff (73.53%). The quality of secondary schools was lower than kindergartens and primary schools in the aspects of room air and surface of object.
Conclusion
The disinfection quality of secondary and primary schools and kindergartens in Wuxi is good in general. More attention should be paid on hand hygiene, concentration of chlorine-containing and ultraviolet lamp. The qualities of room air and surface of object of secondary schools need to be improved.
10.Construction of Clinical Pharmacist Performance Appraisal System under the New “National Examination ” Situation
Yingqi WU ; Shengyu ZHANG ; Tong YIN ; Zhangbao WU ; Bin ZHU ; Aizong SHEN
China Pharmacy 2021;32(18):2184-2189
OBJECTIVE:To prov ide reference for mobilizing the work enthusiasm of clinical pharmacists ,and further promoting the strategic objectives of performance appraisal in three-level public hospitals (“National examination ”for short ). METHODS:A department performance appraisal team was established ,and a key performance indicator system consisting of 4 first-level indicators and 9 second-level indicators was constructed by using literature retrieval and expert consultation. The performance distribution method of double assessment of performance score and performance score was established ,and a performance publicity and feedback performance mechanism was formed. Relevant data were collected to compare the core work indicators of clinical pharmacists ,use intensity of antibiotics ,compliance rate of essential drugs in our hospital from Apr. to Dec. 2019(before implementation )and Apr. to Dec. 2020(after implementation ). RESULTS :After the implementation of performance appraisal scheme ,the total number of medication recommendations of clinical pharmacists increased from 1 192 to 5 226,with an increase of 338.42%;the number of medication suggestions received increased from 846 to 4 855,with an increase of 473.88%; and the rate of drug suggestions received increased from 70.97% to 92.90%;the number of pharmaceutical consultation increased from 195 to 1 284,with an increase of 558.46%;the number of drug counseling increased from 1 203 to 2 719,increasing by 126.02%. Form Apr. to Dec. 2020,the number of patient safety medication evaluation forms reached 660. The antibiotics use density(AUD)in clinical departments of 13 clinical pharmacists were decreased to different extent after the implementation of performance appraisal scheme ,the decine rate was 92.31%(12/13),and the compliance rate was 69.23%(9/13);utilization rate of essential medicine among outpatients of 11 clinical pharmacists ’clinical departments had achieved positive growth ,and those among inpatients of 2 clinical pharmacists ’clinical departments had achieved positive growth. CONCLUSIONS :The performance appraisal system of clinical pharm acists formulated by our hospital links the “National examination ”index with the performance , appraisal of clinical pharmacists ,which can provide ideas for No.2018sxzx57,No.2020jyxm2328,No.2020jyxm2307) the performance appraisal of three-level public hospitals and help to promote the high-quality and sustainable development of hospital pharmaceutical care.


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