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*
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Mice, Inbred BALB C
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Cats
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Mice
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Allergens/genetics*
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Glycoproteins/genetics*
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Enzyme-Linked Immunosorbent Assay
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Hybridomas/immunology*
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Recombinant Proteins/genetics*
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Female
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Antibody Specificity
2.Bioinformatics analysis of a CLCN5 geneframeshift mutation in a patient with Dent disease.
Yingying ZHANG ; Nannan LI ; Liangliang FAN ; Jishi LIU
Journal of Central South University(Medical Sciences) 2025;50(5):913-918
Dent disease is a rare X-linked recessive inherited renal tubular disorder characterized by low molecular weight proteinuria (LMWP), hypercalciuria, nephrocalcinosis, and other clinical features, and can lead to progressive renal failure. It is primarily caused by mutations in the CLCN5 gene. This article reports the case of a 10-year-old male patient of Chinese descent who was incidentally found to have asymptomatic proteinuria during a routine health examination. Comprehensive biochemical testing and clinical evaluation revealed significant LMWP and hypercalciuria, while renal biopsy showed mesangial cell and matrix proliferation. Whole exome sequencing identified a novel deletion mutation in the CLCN5 gene (NM_001127899.4, c.1158delC, p.F387Lfs*42) causing a frameshift and premature termination, which is likely to disrupt its role in chloride/hydrogen ion exchange and endosomal acidification. Bioinformatic analysis indicated the variant is pathogenic. Genetic testing plays an important role in diagnosing rare kidney diseases. Early identification of pathogenic mutations is essential for facilitating timely intervention and appropriate management, potentially enhancing patient outcomes. This report expands the CLCN5 mutation spectrum and contributes to understanding the genetic and molecular mechanisms of Dent disease.
Humans
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Chloride Channels/genetics*
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Dent Disease/genetics*
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Male
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Child
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Computational Biology
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Mutation
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Proteinuria/genetics*
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Hypercalciuria/genetics*
3.Experience of WANG Yongyan in Treating Vascular Cognitive Impairment from the Perspectives of 'Deficient Qi Retention and Stagnation' and 'Toxin Damaging Brain Collaterals'
Yipin FAN ; Jianpeng LI ; Yingying ZHANG ;
Journal of Traditional Chinese Medicine 2025;66(23):2411-2415
This paper summarized the experience of professor WANG Yongyan in differentiating and treating vascular cognitive impairment (VCI). It is believed that the core etiology and pathogenesis of VCI are deficient qi retention and stagnation, and toxin damaging brain collaterals. The essence lies in depletion of essence and qi, and malnutrition of brain collaterals as the root cause, while phlegm, turbidity, stasis, and heat blocking brain collaterals act as the branch aspects. It reveals the pathogenesis essence of VCI as stagnation caused by deficiency and qi stagnation transforming into excess, internal generation of toxin and collateral injury. The overall therapeutic principle is to tonify deficiency, move stagnation, resolve toxin, and unblock the collaterals, guided by the three core treatment methods of nourishing, unblocking, and clearing. Clinically, according to the different characteristics of the plateau phase, fluctuation phase, and decline phase, treatment methods including replenishing qi and blood, tonifying the kidney and supplementing essence; promoting sanjiao (三焦) circulation and activating blood to dredge collaterals; clearing heat and resolving toxin, eliminating phlegm and reducing turbidity are respectively adopted. This provides clinical guidance for the treatment of VCI in traditional Chinese medicine.
4.Research progress in the study of the correlation between oral disease and chronic kidney disease
Yingying FAN ; Zhiyue LU ; Jianqiu JIN
Chinese Journal of Preventive Medicine 2024;58(4):561-568
Chronic kidney disease is a common and serious life-threatening health condition, often associated with multisystemic complications. In recent years, several studies have found that chronic kidney disease is not only closely associated with the occurrence and development of a variety of oral diseases, such as periodontal disease, oral mucosal disease, dental tissue disease, oral bone tissue disease, and oral carcinoma. Meanwhile, chronic kidney disease is also affected by some oral diseases. This article reviews the research progress on the correlation between chronic kidney disease and oral diseases, in order to prevent the occurrence and progression of oral diseases and maintain oral health status. of patients with chronic kidney disease in a more targeted manner.
5.Research progress in the study of the correlation between oral disease and chronic kidney disease
Yingying FAN ; Zhiyue LU ; Jianqiu JIN
Chinese Journal of Preventive Medicine 2024;58(4):561-568
Chronic kidney disease is a common and serious life-threatening health condition, often associated with multisystemic complications. In recent years, several studies have found that chronic kidney disease is not only closely associated with the occurrence and development of a variety of oral diseases, such as periodontal disease, oral mucosal disease, dental tissue disease, oral bone tissue disease, and oral carcinoma. Meanwhile, chronic kidney disease is also affected by some oral diseases. This article reviews the research progress on the correlation between chronic kidney disease and oral diseases, in order to prevent the occurrence and progression of oral diseases and maintain oral health status. of patients with chronic kidney disease in a more targeted manner.
6.Application of patient empowerment in early rehabilitation training for patients with deep vein thrombosis after receiving catheter-directed thrombolysis
Huimin ZHANG ; Shiwu YIN ; Jun CHEN ; Yingying ZHU ; Na ZHANG ; Beibei FAN
Journal of Interventional Radiology 2024;33(5):554-559
Objective To explore the application of patient empowerment in early rehabilitation training for patients with lower extremity deep vein thrombosis(DVT)after receiving catheter-directed thrombolysis(CDT).Methods A total of 110 patients with lower extremity DVT,who were scheduled for CDT therapy,were enrolled in this study.Using random digital table method,the patients were divided into control group(n=55)and intervention group(n=55).Routine rehabilitation activities were implemented for patients of the control group,while patient empowerment-based early rehabilitation training program was carried out for patients of the intervention group.The compliance to rehabilitation exercise,difference of upper/lower patella leg circumference,incidence of complications,duration of catheterization and thrombolysis,and length of hospital stay in both groups were calculated.The Self-Efficacy for Managing Chronic Disease,Chinese version of the Readiness for Hospital Discharge Scale,and the VEnous INsufficiency Epidemiological and Economic Study(VEINES)Quality of Life scale were used to evaluate the rehabilitation results of the patients of two groups.Results There were no significant differences in the duration of catheterization and thrombolysis,incidence of complications,and length of hospital stay between the two groups(all P>0.05).At the time of discharge,the difference of upper/lower patella leg circumference in the intervention group was remarkably smaller than that in the control group,and the compliance with rehabilitation exercise,the self-efficacy,the readiness for discharge,and the quality of life in 3 months after discharge in the intervention group were strikingly better than those in the control group(all P<0.05).Conclusion The implementation of patient empowerment in early rehabilitation training for patients with lower extremity DVT after receiving CDT therapy can improve the compliance with rehabilitation exercise,self-efficacy,limb swelling,readiness for discharge,and quality of life after discharge from hospital.(J Intervent Radiol,2024,33:554-559)
7.Analysis of Clinical Characteristics in 2 Cases of Hypoparathyroidism Sensorineural Deafness and Renal Dysplasia Syndrome
Min LIU ; Liping MENG ; Hui JI ; Ye FAN ; Yingying WANG ; Qin HONG
Journal of Audiology and Speech Pathology 2024;32(5):422-426
Objective To investigate the clinical characteristics and genetic causes in 2 patients with hypopar-athyroidism,sensorineural deafness and renal dysplasia syndrome(HDR).Methods A retrospective analysis of au-diology,gene detection,and other clinical diagnostic data was performed on 2 patients diagnosed with HDR syn-drome.Results Patient 1 failed the newborn hearing screening(otoacoustic emission)and was diagnosed with mod-erate sensorineural hearing loss through audiology evaluation.Follow-up tests of blood calcium and parathyroid hor-mone levels were normal,and ultrasound examinations of the urinary system and parathyroid gland showed no ab-normalities.Patient 2 passed the newborn hearing screening but failed the 3-year-old physical examination(otoa-coustic emission)and was diagnosed with moderate sensorineural hearing loss.Follow-up tests of blood calcium and parathyroid hormone levels were normal,and the parathyroid gland ultrasound showed no abnormalities,but the re-nal ultrasound showed bilateral small renal calculi with normal morphology.Both patients were diagnosed with HDR syndrome through gene testing,and the 2 GAT A3 gene mutation sites(c.867dup,c.65_68dup)causing the disease were both reported for the first time.Conclusion The clinical phenotypes of HDR syndrome are highly variable.Children with suspected hearing loss accompanied by hypoparathyroidism or renal dysfunction should have gene tes-ting and other related examinations as soon as possible to avoid misdiagnosis.
8.Bibliometric analysis of research related to medical humanistic care based
Yingying FAN ; Ziqi LI ; Jin XIE ; Yun LIU
Chinese Medical Ethics 2024;37(5):603-611
Objective:By analyzing the research related to medical humanistic care,the aim is to understand its research status and development trends,and provide theoretical and data support for research in this field.Methods:The relevant literature in the China National Knowledge Infrastructure(CNKI)database were systematically searched.Excel 2019 was used to analyze the situation of annual publication volume,journal distribution,and other content of the literature.CiteSpace software was used to visually analyze the knowledge graph related to authors and keywords.Results:A total of 825 articles were included in the study and the overall number of articles showed an upward trend since 2000.The research hotspots included humanistic care,medical humanities,nursing education,etc.It was found by analyzing that there were three problems in the research of humanistic care in medicine:the research focus is mainly on the need for medical staff to provide patients with more humanistic care,and there is relatively little research on the humanistic care of medical staff themselves;The types of journals are scattered,and the funding support is insufficient;No medical education collaborative research model has been formed.Conclusion:Medical humanistic care is the essence of the professional spirit of physicians and an important indicator in testing the effectiveness of medical humanistic education.In the future,the research on medical humanistic care should strengthen multi-dimensional humanistic care to effectively protect the rights and interests of medical staff,pay more attention to the combination of theoretical research and clinical practice,exploit the instrumental value of narrative medicine,increase support efforts from fund,and improve top-level design.
9.Renal impairment in mice induced by environmental high concentration of polyionized drinking water and high temperature exposure
Yingying LIU ; Fan DING ; Ruojing WANG ; Xuan WU ; Lin ZHANG ; Qing WU
Journal of Environmental and Occupational Medicine 2024;41(5):546-551
Background The burden of chronic kidney diseases (CKD) is continuously increasing in the globe. Environmental factors are one of the trigger factors for chronic kidney diseases of unknown etiology (CKDu). However, the current toxicological evidence on the renal effects induced by environmental high concentrations of multiple ions in drinking water and high temperature exposure is very limited. Objective To preliminary investigate the renal effects of exposure to drinking water with environmental high concentrations of fluoride, calcium, sodium, and bromide ions alone or in combination with high temperature in mice. Methods A mouse drinking water exposure model was established using ICR male mouse (8 weeks old) with exposure to 3 mg·L−1 fluoride ions, 250 mg·L−1 calcium ions, 400 mg·L−1 sodium ions, and 1 mg·L−1 bromide ions (to mimic the high concentration of ions in the groundwater in the areas with a high prevalence rate of CKDu in Sri Lanka) and high temperature of 32 ℃. ICR male mice were randomly divided into a mixed fluoride-calcium-sodium-bromide ion and high temperature exposure group, exposure groups of each ion and high temperature alone, a fluoride-calcium-sodium ion exposure group, and a fluoride-calcium-sodium-bromide ion exposure group. In the control group, the animals were given normal purified water at room temperature of (23±2) ℃. After 12 consecutive weeks of exposure, body weights and liver (kidney) organ coefficients were determined. Assessment of renal histopathologic damage was performed by hematoxylin-eosin staining and pathology scoring. At the end of the 12-week exposure period, 24 h urine samples were collected for the measurements of creatinine (UCr), albumin (ALB), neutrophil gelatinase-associated lipocalin (NGAL), and β2-microglobulin (β2-MG) levels. Cell apoptosis was assessed by TUNEL assay. Results The mice in the mixed exposure group showed a significant decrease in body weight and marked increases in the scores of renal histopathological injuries and the urinary levels of β2-MG compared to those of the control mice (P<0.05). Compared with the control group, the differences in body weight and urinary renal injury indexes of the mice in the fluoride-calcium-sodium and the fluoride-calcium-sodium-bromide ion groups (except for the decrease of the β2-MG levels in urinary in the latter group) were not statistically significant (P>0.05), but the renal histopathological injury scores were significantly increased (P<0.05). By contrast, body weights, liver (kidney) organ coefficient, and renal histopathological injury scores were comparable in the control mice and the mice fed with drinking water containing high levels of a single ion alone or housed at high temperature alone (P>0.05). Furthermore, the renal histopathological injury score showed no significant differences between the fluoride-calcium-sodium ion exposure group and the fluoride-calcium-sodium-bromide ion exposure group (P>0.05). The interaction between bromide ions and fluoride-calcium-sodium ions on renal tissue pathological damage was not statistically significant (P>0.05). Results from the TUNEL assay showed a significant increase in renal cell apoptosis in the fluoride-calcium-sodium ion exposure group (P<0.05). Conclusions Environmental high levels of mixed fluoride, calcium, and sodium ions in drinking water induce renal pathological damage in mice, which are exacerbated in combination with high temperature environment. High temperature exposure alone does not affect the pathological damage of renal tissue,
10.Protective mechanism of rhubarb decoction against inflammatory damage of brain tissue in rats with mild hepatic encephalopathy: A study based on the PI3K/AKT/mTOR signaling pathway
Guangfa ZHANG ; Yingying CAI ; Long LIN ; Lei FU ; Fan YAO ; Meng WANG ; Rongzhen ZHANG ; Yueqiao CHEN ; Liangjiang HUANG ; Han WANG ; Yun SU ; Yanmei LAN ; Yingyu LE ; Dewen MAO ; Chun YAO
Journal of Clinical Hepatology 2024;40(2):312-318
ObjectiveTo investigate the role and possible mechanism of action of rhubarb decoction (RD) retention enema in improving inflammatory damage of brain tissue in a rat model of mild hepatic encephalopathy (MHE). MethodsA total of 60 male Sprague-Dawley rats were divided into blank group (CON group with 6 rats) and chronic liver cirrhosis modeling group with 54 rats using the complete randomization method. After 12 weeks, 40 rats with successful modeling which were confirmed to meet the requirements for MHE model by the Morris water maze test were randomly divided into model group (MOD group), lactulose group (LT group), low-dose RD group (RD1 group), middle-dose RD group (RD2 group), and high-dose RD group (RD3 group), with 8 rats in each group. The rats in the CON group and the MOD group were given retention enema with 2 mL of normal saline once a day; the rats in the LT group were given retention enema with 2 mL of lactulose at a dose of 22.5% once a day; the rats in the RD1, RD2, and RD3 groups were given retention enema with 2 mL RD at a dose of 2.5, 5.0, and 7.5 g/kg, respectively, once a day. After 10 days of treatment, the Morris water maze test was performed to analyze the spatial learning and memory abilities of rats. The rats were analyzed from the following aspects: behavioral status; the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) and the level of blood ammonia; pathological changes of liver tissue and brain tissue; the mRNA and protein expression levels of phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), and mammalian target of rapamycin (mTOR) in brain tissue. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the MOD group, the RD1, RD2, and RD3 groups had a significantly shorter escape latency (all P<0.01), significant reductions in the levels of ALT, AST, IL-1β, IL-6, TNF-α, and blood ammonia (all P<0.05), significant alleviation of the degeneration, necrosis, and inflammation of hepatocytes and brain cells, and significant reductions in the mRNA and protein expression levels of PI3K, AKT, and mTOR in brain tissue (all P<0.05), and the RD3 group had a better treatment outcome than the RD1 and RD2 groups. ConclusionRetention enema with RD can improve cognitive function and inflammatory damage of brain tissue in MHE rats, possibly by regulating the PI3K/AKT/mTOR signaling pathway.

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