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.

Background and purpose:In breast cancer surgery,margin status assessment significantly impacts patient prognosis,with positive margins indicating higher recurrence and metastasis risks.Ensuring complete tumor resection is thus critical for surgical success.Indocyanine green(ICG)has garnered attention for its potential real-time imaging of breast cancer lesions under near-infrared light.This study employed ICG for intraoperative assessment of breast cancer lesion margin status and further explored the possibility of optimizing the safe margin distance surround the lesion in normal breast tissue.Methods:Clinical data of patients admitted to the Department of Thyroid and Breast Surgery,the Fourth Affiliated Hospital of Anhui Medical University(Affiliated Chaohu Hospital),from December 2021 to September 2022 were collected.A retrospective clinical study was conducted on breast cancer patients who were randomly assigned to either the ICG group or the conventional surgery group.Two to three hours before surgery,patients in the ICG group received a peripheral intravenous injection of 0.5 mg/kg ICG.Intraoperative fluorescence imaging was performed on the specimen before and after resection,as well as on the residual cavity.Near-infrared fluorescence imaging equipment was used to quantitatively measure fluorescence intensity of resected lesions at 4 directions(12,3,6,and 9 o'clock)and detect fluorescence in the residual cavity after lesion removal.Specimens were promptly sent to the pathology department for pathological examination,and safety margins of normal breast tissue in the 4 directions were recorded.The Strengthening the Reporting of Observational Studies in Epidemiology(STROBE)checklist was followed for this study.This study was approved by the Ethics Committee of the Fourth Affiliated Hospital of Anhui Medical University(Affiliated Chaohu Hospital)(No.KYXM-202310-46).Results:This study included 50 breast cancer patients,with 24 in the ICG group and 26 in the traditional surgery group.In the ICG group,fluorescence signals were detected at all lesion sites.Specifically,fluorescence density values at the lesion center,margin,and surrounding normal breast tissue were measured as 251.08±10.73,208.08±19.74,and 156.76±16.47,respectively,showing a gradual decrease from center outward with statistically significant differences(P＜0.05).Additionally,fluorescence ratios between the lesion center and margin,and center and surrounding normal tissue,were 1.22±0.13 and 1.62±0.19,respectively.After resection,abnormal fluorescence was observed in 2 of 24 cases in the residual cavity,with 1 case being invasive carcinoma with ductal carcinoma in situ and the other normal breast tissue.Ultimately,this study demonstrated that ICG achieved a sensitivity of 95.9%and a specificity of 97.9%in margin assessment.After specimen resection,the safety margins of normal glandular tissue surrounding the lesion were measured.The safety widths for the ICG group and the concurrent breast cancer surgery group were(8.36±6.42)mm and(15.08±4.75)mm,respectively.This difference was statistically significant(P＜0.05).Conclusion:ICG is a real-time,efficient,and cost-effective tracer that can be used to determine breast cancer margins,with excellent sensitivity and specificity.For early-stage breast cancer patients who are eligible for breast-conserving surgery,this tracer helps to reduce the amount of healthy breast tissue that is removed around the lesion.

Background and purpose:In breast cancer surgery,margin status assessment significantly impacts patient prognosis,with positive margins indicating higher recurrence and metastasis risks.Ensuring complete tumor resection is thus critical for surgical success.Indocyanine green(ICG)has garnered attention for its potential real-time imaging of breast cancer lesions under near-infrared light.This study employed ICG for intraoperative assessment of breast cancer lesion margin status and further explored the possibility of optimizing the safe margin distance surround the lesion in normal breast tissue.Methods:Clinical data of patients admitted to the Department of Thyroid and Breast Surgery,the Fourth Affiliated Hospital of Anhui Medical University(Affiliated Chaohu Hospital),from December 2021 to September 2022 were collected.A retrospective clinical study was conducted on breast cancer patients who were randomly assigned to either the ICG group or the conventional surgery group.Two to three hours before surgery,patients in the ICG group received a peripheral intravenous injection of 0.5 mg/kg ICG.Intraoperative fluorescence imaging was performed on the specimen before and after resection,as well as on the residual cavity.Near-infrared fluorescence imaging equipment was used to quantitatively measure fluorescence intensity of resected lesions at 4 directions(12,3,6,and 9 o'clock)and detect fluorescence in the residual cavity after lesion removal.Specimens were promptly sent to the pathology department for pathological examination,and safety margins of normal breast tissue in the 4 directions were recorded.The Strengthening the Reporting of Observational Studies in Epidemiology(STROBE)checklist was followed for this study.This study was approved by the Ethics Committee of the Fourth Affiliated Hospital of Anhui Medical University(Affiliated Chaohu Hospital)(No.KYXM-202310-46).Results:This study included 50 breast cancer patients,with 24 in the ICG group and 26 in the traditional surgery group.In the ICG group,fluorescence signals were detected at all lesion sites.Specifically,fluorescence density values at the lesion center,margin,and surrounding normal breast tissue were measured as 251.08±10.73,208.08±19.74,and 156.76±16.47,respectively,showing a gradual decrease from center outward with statistically significant differences(P＜0.05).Additionally,fluorescence ratios between the lesion center and margin,and center and surrounding normal tissue,were 1.22±0.13 and 1.62±0.19,respectively.After resection,abnormal fluorescence was observed in 2 of 24 cases in the residual cavity,with 1 case being invasive carcinoma with ductal carcinoma in situ and the other normal breast tissue.Ultimately,this study demonstrated that ICG achieved a sensitivity of 95.9%and a specificity of 97.9%in margin assessment.After specimen resection,the safety margins of normal glandular tissue surrounding the lesion were measured.The safety widths for the ICG group and the concurrent breast cancer surgery group were(8.36±6.42)mm and(15.08±4.75)mm,respectively.This difference was statistically significant(P＜0.05).Conclusion:ICG is a real-time,efficient,and cost-effective tracer that can be used to determine breast cancer margins,with excellent sensitivity and specificity.For early-stage breast cancer patients who are eligible for breast-conserving surgery,this tracer helps to reduce the amount of healthy breast tissue that is removed around the lesion.

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.

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.

Objective To establish an evaluation criterion for high-quality nursing services in comprehensive hospitals of Grade Three in Guangdong Province and provide references for promoting the evaluation of high quality nursing services and standardising high quality nursing services.Methods Based on literature review,brainstorming and references of relevant domestic systems,standards and policies,a pool of evaluation index items and a preliminary questionnaire were constructed.Delphi method was used for two rounds of expert consultations on evaluation indicators.Results The positive coefficients of the two rounds of expert consultations were of 80.00%and 91.67%,respectively,with the authority coefficients both at 0.93.The Kendall coordination coefficients for hospital part were 0.128 and 0.116 respectively,for ward part were 0.193 and 0.107 with statistical differences(both P＜0.001).The evaluation criteria for high-quality nursing services in general hospitals above the third level in Guangdong Province involved in nursing services of hospitals and wards.There were 8 primary indicators,22 secondary indicators and 65 tertiary indicators in the nursing services at hospital part,and 9 primary indicators,23 secondary indicators and 50 tertiary indicators in the nursing services at ward part.Conclusions The evaluation criteria for high-quality nursing services in comprehensive hospitals above the third level in Guangdong Province,established in this study,are scientifically valid and reliable.They allow improvement of management system for the high-quality nursing services as well as guide the healthy development of high-quality nursing services.

Sepsis is characterized by a severe and life-threatening host immune response to polymicrobial infection accompanied by organ dysfunction.Studies on the therapeutic effect and mechanism of immunomod-ulatory drugs on the sepsis-induced hyperinflammatory or immunosuppression states of various im-mune cells remain limited.This study aimed to investigate the protective effects and underlying mechanism of artesunate(ART)on the splenic microenvironment of cecal ligation and puncture-induced sepsis model mice using single-cell RNA sequencing(scRNA-seq)and experimental validations.The scRNA-seq analysis revealed that ART inhibited the activation of pro-inflammatory macrophages recruited during sepsis.ART could restore neutrophils'chemotaxis and immune function in the septic spleen.It inhibited the activation of T regulatory cells but promoted the cytotoxic function of natural killer cells during sepsis.ART also promoted the differentiation and activity of splenic B cells in mice with sepsis.These results indicated that ART could alleviate the inflammatory and/or immunosuppressive states of various immune cells involved in sepsis to balance the immune homeostasis within the host.Overall,this study provided a comprehensive investigation of the regulatory effect of ART on the splenic microenvironment in sepsis,thus contributing to the application of ART as adjunctive therapy for the clinical treatment of sepsis.

Tripterygium glycosides tablet(TGT),the classical commercial drug of Tripterygium wilfordii Hook.F.has been effectively used in the treatment of rheumatoid arthritis,nephrotic syndrome,leprosy,Behcet's syndrome,leprosy reaction and autoimmune hepatitis.However,due to its narrow and limited treatment window,TGT-induced organ toxicity(among which liver injury accounts for about 40％of clinical reports)has gained increasing attention.The present study aimed to clarify the cellular and molecular events underlying TGT-induced acute liver injury using single-cell RNA sequencing(scRNA-seq)technology.The TGT-induced acute liver injury mouse model was constructed through short-term TGT exposure and further verified by hematoxylin-eosin staining and liver function-related serum indicators,including alanine aminotransferase,aspartate aminotransferase,alkaline phosphatase and total bilirubin.Using the mouse model,we identified 15 specific subtypes of cells in the liver tissue,including endothelial cells,hepatocytes,cholangiocytes,and hepatic stellate cells.Further analysis indicated that TGT caused a significant inflammatory response in liver endothelial cells at different spatial locations;led to marked inflammatory response,apoptosis and fatty acid metabolism dysfunction in hepatocytes;activated he-patic stellate cells;brought about the activation,inflammation,and phagocytosis of liver capsular macrophages cells;resulted in immune dysfunction of liver lymphocytes;disturbed the intercellular crosstalk in liver microenvironment by regulating various signaling pathways.Thus,these findings elaborate the mechanism underlying TGT-induced acute liver injury,provide new insights into the safe and rational applications in the clinic,and complement the identification of new biomarkers and ther-apeutic targets for liver protection.