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 ; Antibodies, Monoclonal/biosynthesis* ; Mice, Inbred BALB C ; Cats ; Mice ; Allergens/genetics* ; Glycoproteins/genetics* ; Enzyme-Linked Immunosorbent Assay ; Hybridomas/immunology* ; Recombinant Proteins/genetics* ; Female ; Antibody Specificity

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 ; Chloride Channels/genetics* ; Dent Disease/genetics* ; Male ; Child ; Computational Biology ; Mutation ; Proteinuria/genetics* ; Hypercalciuria/genetics*

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.

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.

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,

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.

Objective:To systematically integrate qualitative research on the psychological experience of non-suicidal self-injury (NSSI) in adolescents.Methods:A comprehensive literature search was conducted in databases including China National Knowledge Infrastructure, WanFang data, VIP, China Biology Medicine disc, PubMed, Web of Science, Embase, and Cochrane Library for qualitative studies on the psychological experience of NSSI in adolescents and the search period covered the inception of the databases until June 27, 2023. The methodological quality of the included studies was assessed using the Joanna Briggs Institute Critical Appraisal Checklist for Qualitative Research (2016 edition). The results were integrated using the Meta-synthesis method.Results:A total of 15 studies were included, comprising ten phenomenological studies, one descriptive qualitative study, one mixed-method study, one narrative study, one exploratory study, and one grounded theory study. Thirty-one findings were extracted and synthesized into ten categories, resulting in three main themes: triggers of adolescent NSSI, NSSI as a coping strategy, and the internal conflict and struggle of adolescents engaging in NSSI.Conclusions:Adolescents engaging in NSSI experience complex psychological and sensory responses. Healthcare professionals should closely monitor adolescents' emotional changes, enhance psychological assessments, and guide them in coping with negative emotions effectively to improve their physical and mental health.

Objectives:This study was designed to observe the impact of frailty status and multiple comorbidities on anticoagulation therapy adherence in elderly patients(≥75 years old)with atrial fibrillation. Methods:This retrospective study enrolled 528 patients(≥75 years old)with atrial fibrillation who were admitted to the geriatric department of Peking University Third Hospital from October 2016 to October 2019.A total of 443(83.9%)elderly patients receiving anticoagulation therapy were included,with an average age of(84.72±5.39)years.Thromboembolic and bleeding risk were evaluated by CHA2DS2-VASc and HAS-BLED,ATRIA bleeding score,while the frailty index(FI)and the age-adjusted Charlson comorbidity index(ACCI)were evaluated.Those with FI≥3 and FI between 1～2 were categorized into the frailty group and pre-frailty group.Patients with ACCI≥5 were defined as elderly multiple comorbidity group(high comorbidity burden).Patients who received anticoagulation therapy after discharge were followed up for one year in the outpatient clinic to observe their medication adherence and to find the related reasons. Results:Thromboembolic risk and bleeding scoring were performed in 443 elderly patients,with CHA2DS2-VASc and HAS-BLED scores of 5.61±1.57 and 3.08±0.89,respectively,and ATRIA score was 4.13±1.90.The FI assessment showed that all enrolled patients were in a state of frailty or pre-frailty.The average ACCI was 5.73±1.51.At one year of follow-up,96(21.67%)patients discontinued anticoagulation therapy.Frailty,high comorbidity burden,and high bleeding risk according to ATRIA(ATRIA≥5)were factors affecting medication adherence,with OR values of 1.114(95%CI:1.041-1.205,P=0.003),3.201(95%CI:2.126-4.509,P=0.001),and 1.457(95%CI:1.212-2.075,P=0.024),respectively.Considering non-death endpoint,both the HAS-BLED score(HAS-BLED≥3 分,OR=1.098,95%CI:1.035-1.116,P=0.014)and warfarin(OR=1.015,95%CI:1.001-1.030,P=0.040)were influential factors of medication compliance. Conclusions:The adherence to anticoagulation therapy post-discharge in elderly patients with atrial fibrillation is significantly influenced by frailty status and high comorbidity burden.If terms of non-death endpoint,high bleeding score and use of warfarin are influential factors of medication compliance.

Objective:To understand the security status of newborn fathers and analyze its related factors.Methods:A total of 256 fathers(aged 22-49 years old)of newborns born within 1 week were select-ed.They were assessed with the self-designed general information questionnaire,Parents'Postnatal Sense of Securi-ty(father version),Emotional Inhibition Scale,and Marriage Satisfaction Subscale of Enrich Marital Inventory.Results:The average score of security was(36.0±4.4).Multiple linear regression analysis showed that the security scores were positively correlated with residence area(β=0.35),education level(β=0.25),preterm birth(β=0.11)and the marital satisfaction scores(β=0.25);and were negatively associated with daily crying time(β=-0.28)and the EIS scores(β=-0.18).Conclusion:The sense of security of postpartum fathers is moderate,and it is associated with residence area,education level,preterm birth,marital satisfaction,daily crying time and emotional inhibition.

Objective:To develop a standardized training model incorporating feedback from a digital assessment tool and to evaluate whether the model provides effective training in tooth preparation.Methods:The study was based on the training data of 53 trainees enrolled between February and June 2018 from multiple institutions in China. The trainees were trained in a standardized training unit on the preparation of right maxillary mesial incisors (11 #) for metal ceramic crowns. Three sessions of practice-assessment-feedback before examination were performed in one day. A digital assessment system was used to obtain total and component scores for the preparation as indicators of observation. The scores of three practice sessions and examination were subjected to analysis of variance. Results:The mean total scores before training, after the first training session, after the second training session, after the third training session, and in the examination were (60.53±12.73), (60.12±12.98), (71.25±13.70), (70.70±11.84), and (69.67±12.85), respectively; the overall difference was statistically significant ( F=19.06, P<0.001). Compared to the total scores before training and after the first training session, the total scores after the second and third training sessions and in the examination were significantly increased ( P<0.001, P<0.001). The score deductions for cutting amount and shoulder were similar, with overall significant differences (score deductions for cutting amount, F=16.20, P<0.001; score deductions for shoulder, F=1.45, P=0.032). Compared to before training and after the first training session, the second and third training sessions and the examination showed significant decreases in score deductions for cutting amount and shoulder ( P<0.001, P=0.048). Conclusions:The feedback-based standardized training process established on a digital evaluation system can rapidly enhance the skills of dentists in tooth preparation through immediate and effective feedback and targeted improvements. The training process enables an independent practice mode and optimizes teaching outcomes.