Objective: To establish an accurate and rapid typing method for Rh typing of samples from patients who have received recent blood transfusions by utilizing the difference in osmotic fragility between fresh and old red blood cells. Methods: A lysing solution suitable for destroying old RBCs was prepared. Sixty-one samples collected in our hospital in 2024 with Rh typing of double groups were treated with the lysing solution to remove the old allogeneic red blood cells while preserving the patient's own fresh red blood cells, followed by repeat Rh typing tests. Results: For 61 samples with Rh typing in double groups, 41 were accurately detected identified through the red blood cell lysis method, yielding an identification rate of 67.21%. No significant difference was observed compared to the detection rate of the commonly used capillary centrifugation modified method (χ =0.103, P>0.05). Conclusion: The red blood cell lysis method provides a novel and rapid experimental approach for clinical use in processing Rh-typed samples that are of double groups, thereby offering a basis for Rh compatibility blood transfusion.

BACKGROUND:The Guilu Erxian Glue consists of Testudinis Plastrum,Cornu Cervi,Lycii Fructus,and Ginseng Radix.In earlier clinical observations,it is discovered that using Guilu Erxian Glue to treat patients with liver-kidney deficiency type knee osteoarthritis effectively alleviated knee pain,increased the range of motion,and improved walking ability.However,the exact mechanism by which oral administration of Guilu Erxian Glue can produce local therapeutic effects on the knee joint is still unclear.OBJECTIVE:To investigate the effects of Guilu Erxian Glue on gut microbiota in rats with knee osteoarthritis and to evaluate its mechanism using 16S rDNA sequencing and machine learning analysis.METHODS:Totally 18 female SD rats were randomly divided into three groups:blank group,model group,and Guilu Erxian Glue group,with 6 rats in each group.A knee osteoarthritis model was prepared using the destabilization of the medial meniscus surgical method.After successful modeling,the Guilu Erxian Glue group was given a decoction of Guilu Erxian Glue by gavage,while the blank and model groups were given an equal amount of distilled water.After 28 days of continuous intervention,high performance liquid chromatography was used to detect the active ingredients of Guilu Erxian Glue.MRI imaging was used to observe the condition of rat knee articular cartilage.Fecal samples were collected;DNA was extracted using a kit,amplified and purified by PCR,and an Illumina sequencing library was constructed.The Illumina MiSeq platform was used for high-throughput sequencing to generate raw sequence data.After obtaining the raw data,QIIME2 software was used to process the data.Linear Discriminant Analysis Effect Size analysis and random forest algorithm were used to screen for differential species in microbial data.KEGG and MetaCyc functional pathway analyses were used to explore the association between key microbial communities and experimental groups.Linear discriminant analysis effect values and random forest algorithm were used to screen for differential species.Association networks were used to analyze the interactions between microbial communities,and machine learning methods were used to analyze the composition and changes of gut microbiota.RESULTS AND CONCLUSION:(1)LC-MS component identification was conducted on the traditional Chinese medicine formula of Guilu Erxian Glue,and a total of 7 effective ingredients were identified.(2)MRI imaging showed that synovitis scope of high-density shadows in rats of the Guilu Erxian Glue group was reduced,and the degeneration of medial femoral condyle cartilage was less than that in the model group.(3)16S rDNA sequencing showed that the model group rats exhibited significant microbial imbalance,with a significant decrease in the abundance of Firmicutes and Bacteroidetes at the phylum level,while the proportion of Proteobacteria increased significantly(P＜0.05).The gut microbiota structure of rats in the Guilu Erxian Glue group was significantly improved,and the proportion of Firmicutes and Bacteroidetes increased,restoring a more diverse microbiota composition,approaching that of the blank group(P＜0.05).(4)KEGG and MetaCyc functional pathway analysis showed that the Guilu Erxian Glue group significantly activated multiple metabolic pathways,including amino acid metabolism,lipid metabolism,and biotin synthesis pathways(P＜0.05).(5)The results indicate that Guilu Erxian Glue contains seven active ingredients,and the changes in gut microbiota of knee osteoarthritis rats were analyzed using 16S rDNA sequencing.Guilu Erxian Glue can significantly improve the imbalance of gut microbiota,restore the abundance of beneficial bacteria,and have a significant impact on the composition of gut microbiota,providing scientific basis for the efficacy and mechanism of Guilu Erxian Glue.

BACKGROUND:The Guilu Erxian Glue consists of Testudinis Plastrum,Cornu Cervi,Lycii Fructus,and Ginseng Radix.In earlier clinical observations,it is discovered that using Guilu Erxian Glue to treat patients with liver-kidney deficiency type knee osteoarthritis effectively alleviated knee pain,increased the range of motion,and improved walking ability.However,the exact mechanism by which oral administration of Guilu Erxian Glue can produce local therapeutic effects on the knee joint is still unclear.OBJECTIVE:To investigate the effects of Guilu Erxian Glue on gut microbiota in rats with knee osteoarthritis and to evaluate its mechanism using 16S rDNA sequencing and machine learning analysis.METHODS:Totally 18 female SD rats were randomly divided into three groups:blank group,model group,and Guilu Erxian Glue group,with 6 rats in each group.A knee osteoarthritis model was prepared using the destabilization of the medial meniscus surgical method.After successful modeling,the Guilu Erxian Glue group was given a decoction of Guilu Erxian Glue by gavage,while the blank and model groups were given an equal amount of distilled water.After 28 days of continuous intervention,high performance liquid chromatography was used to detect the active ingredients of Guilu Erxian Glue.MRI imaging was used to observe the condition of rat knee articular cartilage.Fecal samples were collected;DNA was extracted using a kit,amplified and purified by PCR,and an Illumina sequencing library was constructed.The Illumina MiSeq platform was used for high-throughput sequencing to generate raw sequence data.After obtaining the raw data,QIIME2 software was used to process the data.Linear Discriminant Analysis Effect Size analysis and random forest algorithm were used to screen for differential species in microbial data.KEGG and MetaCyc functional pathway analyses were used to explore the association between key microbial communities and experimental groups.Linear discriminant analysis effect values and random forest algorithm were used to screen for differential species.Association networks were used to analyze the interactions between microbial communities,and machine learning methods were used to analyze the composition and changes of gut microbiota.RESULTS AND CONCLUSION:(1)LC-MS component identification was conducted on the traditional Chinese medicine formula of Guilu Erxian Glue,and a total of 7 effective ingredients were identified.(2)MRI imaging showed that synovitis scope of high-density shadows in rats of the Guilu Erxian Glue group was reduced,and the degeneration of medial femoral condyle cartilage was less than that in the model group.(3)16S rDNA sequencing showed that the model group rats exhibited significant microbial imbalance,with a significant decrease in the abundance of Firmicutes and Bacteroidetes at the phylum level,while the proportion of Proteobacteria increased significantly(P＜0.05).The gut microbiota structure of rats in the Guilu Erxian Glue group was significantly improved,and the proportion of Firmicutes and Bacteroidetes increased,restoring a more diverse microbiota composition,approaching that of the blank group(P＜0.05).(4)KEGG and MetaCyc functional pathway analysis showed that the Guilu Erxian Glue group significantly activated multiple metabolic pathways,including amino acid metabolism,lipid metabolism,and biotin synthesis pathways(P＜0.05).(5)The results indicate that Guilu Erxian Glue contains seven active ingredients,and the changes in gut microbiota of knee osteoarthritis rats were analyzed using 16S rDNA sequencing.Guilu Erxian Glue can significantly improve the imbalance of gut microbiota,restore the abundance of beneficial bacteria,and have a significant impact on the composition of gut microbiota,providing scientific basis for the efficacy and mechanism of Guilu Erxian Glue.

Objective To investigate clinical features and severity influencing factors of misophonia in psychiatric outpatients. Methods A total of 130 misophonia patients from Weifang People's Hospital (Oct 2023-Jun 2025) were assessed with scales and psychiatric interviews. Linear regression was used to analyze influencing factors. Results Among the 52.3% reported gradually worsening symptoms. Primary triggers were human oronasopharyngeal sounds (65.3%). Core emotional responses were disgust (86.2%) and anger (61.5%), with 75.4% experiencing loss of control. Common coping strategies included covering ears/wearing earplugs (66.1%) and avoidance (64.6%). Mean Amsterdam Misophonia Scale-Revised score was 26.4±5.2, indicating severe social dysfunction. Common comorbidities were obsessive-compulsive disorder (11.5%), depressive disorder (8.5%), and social anxiety disorder (5.4%). Multivariate analysis identified depressive disorder, social anxiety disorder, obsessive-compulsive disorder and post-traumatic stress disorder as independent factors affecting severity (all P<0.05). Conclusion Misophonia follows a chronic course with severe impairment in social functioning and quality of life. Obsessive-compulsive disorder, depressive disorder, social anxiety disorder, and post-traumatic stress disorder are key factors influencing symptom severity.

Objective: To eliminate the interference of daratumumab (DARA) in the titer determination of IgG antiA/antiB antibodies, resolve the discrepancy between test results and theoretical expectations, and obtain accurate and reliable test results. Methods: A group A blood sample with a positive IgG antiA titer result was confirmed to have a recent history of DARA administration. After removal of IgM antibodies from the plasma, IgG antiA/antiB titers were determined using the antiglobulin test card with a serial twofold dilution method, following pretreatment of reagent red blood cells with a rabbit antihuman CD38 functional monoclonal antibody. Results: Without pretreatment with the rabbit antihuman CD38 functional monoclonal antibody, the IgG antiA titer of the sample was 128. After pretreatment, the IgG antiA titer became negative. Conclusion: Based on the red blood cell CD38 antigen masking strategy, pretreatment of red blood cells with a rabbit antihuman CD38 functional monoclonal antibody can eliminate the interference of DARA on the titer determination of IgG antibodies, thereby providing accurate diagnostic evidence and decision support for clinical treatment.

The aim of this study is to establish a rapid,efficient,and sensitive method for detecting the infectious laryngotracheitis virus(ILTV).The DNA of ILTV was extracted and used as a tem-plate to develop a recombinant enzyme-mediated isothermal amplification(RAA)fluorescence de-tection method for ILTV through optimization of conditions,sensitivity analysis,and repeatability assessment.Additionally,the nucleic acids of avian influenza virus(AIV),IBV,and Newcastle dis-ease virus(NDV)were detected to verify the specificity of this method.Finally,this method was applied to analyze 59 clinical samples collected from multiple large-scale chicken farms in Hebei Province,and the results were compared with those obtained from real-time fluorescence quantifi-cation(qPCR)and PCR methods according to national standards.The results showed that the RAA detection method established in this study had a reaction system of 25.0 μL buffer,2.1 μL primer,0.6 μL probe,5.0 μL magnesium acetate,and 5.0 μL template.The reaction temperature was 39 ℃ and the amplification time was within 20 minutes.The sensitivity of this method was 101 copies/μL,and the specificity detection was 100％.Testing of 59 clinical samples showed that 17 were detected positive by both RAA fluorescence and qPCR,and 12 were detected by PCR,and the detection rate of RAA(fluorescence)was consistent with real-time fluorescence quantification and qPCR,which was higher than that of the PCR assay.The research results indicate that the RAA fluorescence method has a short detection time,good specificity and sensitivity,and can be used for rapid detection of ILTV.

Objective:To investigate and analyze the service needs of family members of Neurosurgical Intensive Care Unit patients and provide references for improving Neurosurgical Intensive Care Unit nursing services.Methods:Totally 270 family members of neurosurgical patients hospitalized between March and May 2024 at Intensive Care Unit of Beijing Tiantan Hospital, Capital Medical University were selected by convenience sampling. Kano Model-based Questionnaire on the Needs of Family Members of Neurosurgical Intensive Care Unit Patients designed based on literature review and expert consultation, was distributed to investigate their families service needs. The Kano model analysis was applied to categorize service needs and prioritize them, followed by constructing a satisfaction-importance quadrant diagram.Results:A total of 270 questionnaires were distributed, with 248 valid responses collected, yielding an effective response rate of 91.85% (248/270) . The Kano attribute analysis revealed that all service needs were classified as "expected attributes". The satisfaction-importance quadrant diagram indicated that 10 items were located in the "advantage zone", nine items in the "retention zone", four items in the "observation zone", and seven items in the "improvement zone".Conclusions:The Kano model-based analysis of service needs helps healthcare providers identify the diverse needs of family members of Neurosurgical Intensive Care Unit patients. It is recommended that while maintaining the provision of services in the "advantage zone", healthcare providers integrate hospital resources to continuously improve services in the "improvement zone" to enhance family satisfaction and strengthen hospital competitiveness.

The aim of this study is to establish a rapid,efficient,and sensitive method for detecting the infectious laryngotracheitis virus(ILTV).The DNA of ILTV was extracted and used as a tem-plate to develop a recombinant enzyme-mediated isothermal amplification(RAA)fluorescence de-tection method for ILTV through optimization of conditions,sensitivity analysis,and repeatability assessment.Additionally,the nucleic acids of avian influenza virus(AIV),IBV,and Newcastle dis-ease virus(NDV)were detected to verify the specificity of this method.Finally,this method was applied to analyze 59 clinical samples collected from multiple large-scale chicken farms in Hebei Province,and the results were compared with those obtained from real-time fluorescence quantifi-cation(qPCR)and PCR methods according to national standards.The results showed that the RAA detection method established in this study had a reaction system of 25.0 μL buffer,2.1 μL primer,0.6 μL probe,5.0 μL magnesium acetate,and 5.0 μL template.The reaction temperature was 39 ℃ and the amplification time was within 20 minutes.The sensitivity of this method was 101 copies/μL,and the specificity detection was 100％.Testing of 59 clinical samples showed that 17 were detected positive by both RAA fluorescence and qPCR,and 12 were detected by PCR,and the detection rate of RAA(fluorescence)was consistent with real-time fluorescence quantification and qPCR,which was higher than that of the PCR assay.The research results indicate that the RAA fluorescence method has a short detection time,good specificity and sensitivity,and can be used for rapid detection of ILTV.

Objective:To investigate and analyze the service needs of family members of Neurosurgical Intensive Care Unit patients and provide references for improving Neurosurgical Intensive Care Unit nursing services.Methods:Totally 270 family members of neurosurgical patients hospitalized between March and May 2024 at Intensive Care Unit of Beijing Tiantan Hospital, Capital Medical University were selected by convenience sampling. Kano Model-based Questionnaire on the Needs of Family Members of Neurosurgical Intensive Care Unit Patients designed based on literature review and expert consultation, was distributed to investigate their families service needs. The Kano model analysis was applied to categorize service needs and prioritize them, followed by constructing a satisfaction-importance quadrant diagram.Results:A total of 270 questionnaires were distributed, with 248 valid responses collected, yielding an effective response rate of 91.85% (248/270) . The Kano attribute analysis revealed that all service needs were classified as "expected attributes". The satisfaction-importance quadrant diagram indicated that 10 items were located in the "advantage zone", nine items in the "retention zone", four items in the "observation zone", and seven items in the "improvement zone".Conclusions:The Kano model-based analysis of service needs helps healthcare providers identify the diverse needs of family members of Neurosurgical Intensive Care Unit patients. It is recommended that while maintaining the provision of services in the "advantage zone", healthcare providers integrate hospital resources to continuously improve services in the "improvement zone" to enhance family satisfaction and strengthen hospital competitiveness.

Objective To determine the reference red blood cells with weak agglutination intensity of low positive quali-ty control products by comparing RhD antigen expression intensity difference according to the serological results.Methods The RhD(+)red blood cells were detected by microcolumn gel method with 1 500 times diluted anti-D typing reagent.The samples with weak and strong RhD antigen expression intensity were selected as the reference red blood cells for weak agglu-tination intensity of low positive quality control products,and verification was performed.Results Ten RhD(+)red blood cells were detected with diluted anti-D typing reagent,of which 8 were 1+and 2 were±.Red blood cells with agglutination intensity of 1+were used as the benchmark to determine the maximum dilution ratio of anti-D typing reagent when their ag-glutination intensity was 1+.As the preparation standard of low positive quality control products,the agglutination intensity of red blood cells with low RhD antigen expression intensity was extremely weak±,which was difficult to ensure the stability of its control limit properties.Based on red blood cells with agglutination intensity of±,the maximum dilution ratio of anti-D typing reagent with agglutination intensity of 1+was re-determined as the preparation standard of low positive quality con-trol products,and the results met the requirements of quality control product setting.Conclusion Using red blood cells with low RhD antigen expression intensity as the benchmark to set the weak agglutination intensity of the low positive quality control products can avoid the loss of control due to the low target value.