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

Debates persist regarding the efficacy and safety of azvudine, particularly its real-world outcomes. This study involved patients aged ≥60 years who were admitted to 25 hospitals in mainland China with confirmed SARS-CoV-2 infection between December 1, 2022, and February 28, 2023. Efficacy outcomes were all-cause mortality during hospitalization, the proportion of patients discharged with recovery, time to nucleic acid-negative conversion (T _NANC), time to symptom improvement (T _SI), and time of hospital stay (T _HS). Safety was also assessed. Among the 5884 participants identified, 1999 received azvudine, and 1999 matched controls were included after exclusion and propensity score matching. Azvudine recipients exhibited lower all-cause mortality compared with controls in the overall population (13.3% vs. 17.1%, RR, 0.78; 95% CI, 0.67-0.90; P = 0.001) and in the severe subgroup (25.7% vs. 33.7%; RR, 0.76; 95% CI, 0.66-0.88; P < 0.001). A higher proportion of patients discharged with recovery, and a shorter T _NANC were associated with azvudine recipients, especially in the severe subgroup. The incidence of adverse events in azvudine recipients was comparable to that in the control group (2.3% vs. 1.7%, P = 0.170). In conclusion, azvudine showed efficacy and safety in older patients hospitalized with COVID-19 during the SARS-CoV-2 omicron wave in China.

ObjectiveTo investigate the effects of Guizhi Fulingwan on oxidative stress and renal fibrosis in diabetic mice based on the nuclear factor E₂-associated factor 2/antioxidant response element （Nrf2/ARE） pathway， and to explore its mechanism of action. MethodA diabetic mouse model was established by feeding high-fat diet combined with intraperitoneal injection of streptozotocin. The diabetic mice were divided into a model group， a metformin group （250 mg·kg^-1）， and Guizhi Fulingwan low， medium， and high dose groups （400.4， 800.8， 1 601.6 mg·kg^-1）， with 10 mice in each group. Another 10 mice were included in a normal group. The treatment was conducted continuously for 8 weeks. Body mass and blood glucose were recorded， and the renal index was calculated. Triglyceride （TG）， total cholesterol （TC）， urea nitrogen （BUN）， serum creatinine （SCr）， superoxide dismutase （SOD）， and malondialdehyde （MDA） levels were measured. Pathological changes in renal tissues were observed using Hematoxylin-eosin， periodate-Schiff， and Masson staining. The protein levels of α-smooth muscle actin （α-SMA）， fibronectin （FN）， and transforming growth factor-β₁ （TGF-β₁） were detected by immunohistochemistry. The protein levels of Nrf2， Kelch-like ECH-associated protein 1 （Keap1）， SOD1， NAD（P）H single-nucleotide oxidase 1 （NQO1）， and heme oxygenase-1 （HO-1） were detected by Western blot. ResultCompared with the normal group， the model group showed elevated glucose， renal index， TG， TC， BUN， and SCr levels， thickened glomerular basement membrane， increased collagen fibers in renal tissues， decreased SOD activity， elevated MDA content， increased protein levels of FN， TGF-β₁， α-SMA， and Keap1， and decreased protein levels of Nrf2， SOD1， NQO1， and HO-1 （P<0.01）. Compared with the model group， the Guizhi Fulingwan and metformin groups showed reduced blood glucose， renal index， TG， TC， BUN， and SCr levels， improved renal histopathological injury， significantly increased SOD activity， significantly reduced MDA content， reduced protein levels of FN， TGF-β₁， α-SMA， and Keap1， and increased protein levels of Nrf2， SOD1， NQO1， and HO-1 （P<0.05， P<0.01）. ConclusionGuizhi Fulingwan can improve oxidative stress and inhibit renal fibrosis， and its mechanism may be related to the activation of the Nrf2/ARE signaling pathway.

Objective:To construct an anticipatory grief intervention scheme for family caregivers of advanced cancer patients based on narrative theory, and to provide reference for anticipatory grief nursing intervention.Methods:From October 2022 to May 2023, through literature research, semi-structured interview and brainstorming method, the first draft of nursing intervention plan was constructed, the Delphi method was used to conduct 2 rounds of correspondence consultation with 15 experts, and the indicators at all levels were modified according to the opinions of experts, and the final draft of intervention plan was formed.Results:The experts were all female, aged (49.67 ± 5.83) years old. The authority coefficient of the two rounds of experts was 0.87. The Kendall coordination coefficients of the first, second, and third level indicators after the first round of expert inquiry were 0.195, 0.113, and 0.093, respectively. The Kendall coordination coefficients of the first, second, and third level indicators after the second round of expert inquiry were 0.200, 0.119, and 0.101, respectively. The differences were statistically significant ( χ2 values were 8.76-107.21, all P<0.05).Finally, a nursing intervention plan based on narrative theory was formed, which included 4 primary indicators, 19 secondary indicators and 72 tertiary indicators. Conclusions:The anticipatory grief intervention scheme for family caregivers of advanced cancer patients is scientific, practical and feasible, and can be used for psychological nursing of family caregivers.

Objective To explore the feasibility and accuracy of neutrophil-to-lymphocyte ratio(NLR)in the prediction of testicular torsion(TT)in children with acute scrotal pain.Methods A retrospective case-control study was performed on 158 pediatric patients with ultrasound suspicion of TT who underwent surgical testicular examination during Jan.2017 and Jan.2024.The patients were divided into TT group and non-TT group.Clinical data and laboratory data at admission were analyzed.Sensitivity and specificity of NLR to TT were determined with the area under the curve(AUC)represented on the receiver operating characteristic(ROC)curves.Results There were with no statistically significant differences in clinical data between the two groups(P＞0.05).The NLR was significantly higher in the TT group than in the non-TT group[(4.82±2.37)vs.(2.85±0.75),P＜0.05].The optimal cut-off value of TT predicted by NLR was 2.07,the AUC was 0.809(95％CI:0.709-0.909),and the sensitivity and specificity were 97.9％and 93.3％,respectively,which were significantly higher than other factors.Conclusion For suspicious ultrasound diagnosis of pediatric acute scrotal pain cases,NLR can be used to predict the possibility of TT and may help to evaluate the urgent surgical treatment in these patients.

Objective:To explore the influencing factors of anticipatory grief among family caregivers of terminal-stage cancer patients,and provide a reference for formulating intervention programs for anticipatory grief.Methods:An interview outline was designed,and the descriptive qualitative research method was used.The aim was to select 11 family narrative caregivers of patients in the Department of Medical Oncology,Nanfang Hospital,Southern Medical University for semi-structured in-depth interviews.Based on the social ecology model,using the narrative theory of life and health as the framework,the content analysis method was adopted to analyze the data.Results:A total of 3 themes and 12 sub-themes were extracted,including the influencing factors at the individual level(narrative literacy,cognitive level towards death,and sense of self-worth),the influencing factors at the interpersonal level(patient's disease situation,education level,economic status,family narrative connection,peer narrative connection,and narrative connection between doctors and family members),and the influencing factors at the cultural policy level(social culture,physical environment of the ward,and policy level).Conclusion:Anticipatory grief was affected by multiple factors,and multi-dimensional narrative connection should be constructed from individual,interpersonal,family,social,and other levels to prevent caregivers from falling into a narrative lock of single caregiver identity,alleviate the impact of anticipatory grief on caregivers,and improve their quality of life.

With the development of artificial intelligence(AI),AI ethics has become one of the core topics of discussion and research.Starting from the ethical issues and human crises that may be triggered by AI in the process of medical application,this paper discussed the significance of the interpersonal narrative connection between doctors and patients advocated by narrative medicine.It was proposed that to ensure the ethicality,deep humanity,and deep emotional connection of AI applications,AI should create a"life narrative database"based on the"human life database"in the process of basic database modelling.Meanwhile,it also revealed that Chinese narrative medicine helped to improve the professional narrative ability of medical doctors and realise deeply humanized medicine,while actively promoting human-machine value symbiosis and contributing narrative power to the deep humanisation of AI.Combining the concept of narrative medicine in China with the future development and application prospects of medical AI,on the one hand,aims to raise awareness among more people about the importance of interpersonal narrative connection and narrative data for medical quality,on the other hand,calls for active cultivation of clinical narrative thinking by both real doctors and future intelligent doctors to better realize the bioethical and humanistic values emphasized by big health.

Objective:To explore the relationship between hemoglobin variability (Hb-var) and risk of all-cause death and cardiovascular death in patients with peritoneal dialysis (PD), and to provide basis for reducing the risk of death in PD patients.Methods:It was a multicenter retrospective cohort study. The clinical data of regular PD patients from Nanfang Hospital of Southern Medical University, Shunde Hospital of Southern Medical University, Foshan First People's Hospital and Ganzhou People's Hospital from July 1, 2008 to December 31, 2019 were collected. Hb-var was calculated based on hemoglobin at baseline before PD and in the first year after PD. The patients were divided into low Hb-var group, moderate Hb-var group and high Hb-var group according to the tertiles of first year Hb-var, and the differences of baseline clinical data among three groups were compared. Follow-up endpoints included death, transfer to hemodialysis, transfer to kidney transplantation, transfer to other centers, loss of follow-up, or on December 31, 2021. Cox regression analysis model was used to analyze the association of the first-year Hb-var with all-cause death and cardiovascular death. Fine-Gray competitive risk regression model was used to evaluate the impact of competitive events on mortality risk.Results:A total of 1 562 patients with PD were included in the study, aged (47.6±13.8) years old, with 821 males (52.6%) and baseline hemoglobin of 81 (69, 94) g/L. Hb-var in the first year of PD was 26.6 (16.7, 40.3) g/L. There were statistically significant differences in age, body mass index, serum albumin, hemoglobin, serum creatinine, serum calcium, serum phosphorus, intact parathyroid hormone and the proportion of renin-angiotensin system inhibitors among low Hb-var group (<20.0 g/L), moderate Hb-var group (20.0-35.5 g/L) and high Hb-var group (≥35.5 g/L, all P<0.05). The follow-up time was 33 (19, 51) months, and 208 patients (13.3%) died, among which 111 patients (53.4%) died of cardiovascular death. Multivariate Cox regression analysis showed that the higher Hb-var in the first year, the lower the risk of all-cause death ( HR=0.98, 95% CI 0.97-0.99, P=0.018) and cardiovascular death ( HR=0.98, 95% CI 0.97-0.99, P=0.041) in PD patients. Compared with low Hb-var group, the risk of all-cause death ( HR=0.56, 95% CI 0.37-0.82, P=0.003) and cardiovascular death ( HR=0.54, 95% CI 0.31-0.95, P=0.032) was lowest in the high Hb-var group. The competitive risk regression model analysis showed that Hb-var in the first year was still negatively correlated with the risk of all-cause death ( HR=0.98, 95% CI 0.97-0.99, P=0.041) and cardiovascular death ( HR=0.98, 95% CI 0.97-0.99, P=0.039). Conclusion:High Hb-var in the first year is associated with low risk of all-cause death and cardiovascular death in PD patients with severe anemia at baseline.

Objective:To describe the incidence of peritoneal dialysis (PD) catheter exit site infection (ESI) and to analyze its pathogenic bacteria characteristics.Methods:The clinical data of PD patients with ESI in Nanfang Hospital of Southern Medical University from 2020 to 2022 was reviewed to describe the dynamic changes in the incidence of ESI, and to analyze the distribution of ESI pathogens, antimicrobial sensitivity analysis of topical antibiotics and treatment outcomes.Results:A total of 187 ESI episodes occurred in 159 PD patients. The proportion of gram-positive and gram-negative ESI were 80.7% and 18.3%, respectively. The main gram-positive and gram-negative pathogen were coagulase negative Staphylococcus (51.1%) and Pseudomonas aeruginosa (60.0%). Rifampicin (sensitivity rate of gram-positive organisms was 88.9%; gram-negative organisms was 100.0%) and gentamicin (sensitivity rate of gram-positive organisms was 83.6%; gram-negative organisms was 93.3%) is the main topical sensitive antibiotics. 137 cases (73.3%) of ESI were cured within 2 weeks after treatment. Among the 50 cases (26.7%) of refractory ESI, 22.0% of cases were associated with tunnel infection, 10.0% with tunnel reconstruction, and only 4.0% with removal of PD catheter. Conclusion:Monitoring and reporting of ESI is an important part to improve the quality of PD management. By reviewing the incidence of ESI, pathogenic bacteria characteristics and treatment effects, real and reliable observational data for the formulation of relevant guidelines and clinical diagnosis and treatment can be provided.

AIM:To investigate the effects of ginkgetin on neurological deficit and angiogenesis induced by cerebral ischemia/reperfusion(I/R)and its underlying mechanism.METHODS:Sixty SD rats were randomly allocated into three groups:sham group,I/R model group,and ginkgetin(40 mg/kg)treatment(I/R+ginkgetin)group,with twenty rats in each group.The middle cerebral artery occlusion/reperfusion(MCAO/R)rat model was employed to simulate cere-bral I/R,and ginkgetin was administered continuously for 7 days following reperfusion.The cerebral infarction volume was quantified using TTC staining.Neuronal density in the ischemic penumbra was assessed through Nissl staining and immu-nohistochemistry for neuron-specific nuclear protein(NeuN).Microvessel density and angiogenesis in the ischemic pen-umbra of each group were analyzed using CD31 labeling and BrdU/von willebrand factor(vWF)double labeling immuno-fluorescence staining.Western blot analysis was performed to determine the levels of heat shock protein 70(HSP70),vas-cular endothelial growth factor(VEGF),and hypoxia inducible factor-1α(HIF-1α)in the ischemic penumbra.RE-SULTS:Compared with the I/R model group,the cerebral infarction volume was significantly reduced in ginkgetin treat-ment group(P<0.01),the number of neurons,the microvessel density,angiogenesis and the expression levels of HIF-1α,VEGF,and HSP70 in the ischemic penumbra were significantly increased(P<0.01).CONCLUSION:Ginkgetin exhibits the potential to augment angiogenesis and facilitate neurological function recovery in MCAO rats,while concur-rently upregulating the expression of HSP70,VEGF,and HIF-1α within the ischemic penumbra.