Hemoglobin, the principal protein in red blood cells, is crucial for oxygen transport in the bloodstream. The quantification of hemoglobin concentration is indispensable in medical diagnostics and health management, which encompass the diagnosis of anemia and the screening of various blood disorders. Immunological methods, based on antigen-antibody interactions, are distinguished by their high sensitivity and accuracy. Consequently, it is necessary to develop hemoglobin-specific antibodies characterized by high specificity and affinity to enhance detection accuracy. In this study, we immunized a Bactrian camel (Camelus bactrianus) with human hemoglobin and subsequently constructed a nanobody library. Utilizing a solid-phase screening method, we selected nanobodies and evaluated the binding activity of the screened nanobodies to hemoglobin. Initially, human hemoglobin was used to immunize a Bactrian camel. Following four immunization sessions, blood was withdrawn from the jugular vein, and a nanobody library with a capacity of 2.85×10⁸ colony forming units (CFU) was generated. Subsequently, ten hemoglobin-specific nanobody sequences were identified through three rounds of adsorption-elution-enrichment assays, and these nanobodies were subjected to eukaryotic expression. Finally, enzyme-linked immunosorbent assay and biolayer interferometry were employed to evaluate the stability, binding activity, and specificity of these nanobodies. The results demonstrated that the nanobodies maintained robust binding activity within the temperature range of 20-40 ℃ and exhibited the highest binding activity at pH 7.0. Furthermore, the nanobodies were capable of tolerating a 10% methanol solution. Notably, among the nanobodies tested, VHH-12 displayed the highest binding activity to hemoglobin, with a half maximal effective concentration (EC₅₀) of 10.63 nmol/L and a equilibrium dissociation constant (K_D) of 2.94×10^-7 mol/L. VHH-12 exhibited no cross-reactivity with a panel of eight proteins, such as ovalbumin and bovine serum albumin, while demonstrating partial cross-reactivity with hemoglobin derived from porcine, goat, rabbit, and bovine sources. In this study, a hemoglobin-specific high-affinity nanobody was successfully isolated, demonstrating potential applications in disease diagnosis and health monitoring.
Animals ; Camelus/immunology* ; Single-Domain Antibodies/immunology* ; Hemoglobins/immunology* ; Humans ; Peptide Library

Objective To investigate the diagnostic value of our regression model based on quantitative parameters of dual-energy CT and clinical features for stages T2 and T3 colorectal cancer.Methods A cross-section study was performed on 91 patients with colorectal cancer confirmed by postoperative pathology in our hospital from January 2022 to November 2023.All of them underwent dual-energy CT examination.According to the pathological T staging criteria of Chinese Colorectal Cancer Diagnosis and Treatment Standard(2020 Edition),they were divided into T2 group(n=43)and T3 group(n=48).Univariate analysis was used to compare the differences in quantitative CT parameters and clinical features between the 2 groups,and the obtained significant variables were employed to construct diagnosis models by univariate or multivariate logistic regression analysis.The area under receiver operating characteristic curve(AUC)of the CT parametric model and the model combined with clinical features was compared to evaluate the efficacy of diagnosing T2 and T3 stages.Results Univariate analysis showed that carcinoembryonic antigen(CEA),N stage,tumor location,tumor longest diameter(LD),CT value of virtual noncontrast(CT-VNC),fat fraction,electron density(Rho)and dual energy index(DEI)were significantly different between the T2 and T3 groups(P＜0.05).Multivariate logistic regression analysis found that N stage,tumor location,LD,fat fraction and DEI were independent risk factors for the diagnosis of stage T3.The AUC value of the model of above CT parameters in diagnosing stage T3 colorectal cancer was 0.671(95％CI:0.558～0.783),and the AUC value of the combined model of above CT parameters and clinical features was 0.886(95％CI:0.815～0.957),and statistical difference was observed in the AUC value between the combined model and the CT parametric model(P＜0.01).Conclusion The regression model constructed with dual-energy CT quantitative parameters combined with clinical features has high value in the preoperative diagnosis of stages T2 and T3 colorectal cancer before surgery.

Objective:The differentially expressed genes(DEGs)and activated signaling pathways in systemic sclerosis(SSc)were screened by bioinformatics methods,and Chinese medicines for potential treatment of SSc were explored,providing a new theoretical basis for the study of SSc and the screening of potential markers.Methods:The data sets GSE58095,GSE130953,GSE33463 and GSE58613 were selected from GEO database and divided into skin group and peripheral blood group according to the sample source.The DEGs of SSc patients was analyzed by R language,and the Wayne diagram was drawn to take the intersection of the two groups.Metascape was used for GO enrichment analysis and KEGG pathway enrichment analysis,and STRING and Cytoscape were used for protein interaction network analysis to find key pathways and hub genes.The core genes were mapped to the medical on-tology information retrieval platform,and related Chinese medicines for SSc treatment were screened.The effective components of Chi-nese medicines were obtained through TCMSP and HERB databases,and the target letters of active ingredients were obtained through swiss database.The"drug-active ingredient-target"network was constructed by Cytoscape.Results:Total 218 DEGs were identified from the skin group of patients with SSc,and 283 DEGs were screened from peripheral blood of patients with SSc.Among them,there were 7 DEGs co-upregulated in skin and peripheral blood,namely ISG15,LGALS3BP,BST2,C1QB,IFI27,CEACAM1 and FBP1.CAMK2N1 was up-regulated in skin but down-regulated in peripheral blood,ARG1 was down-regulated in skin but up-regulated in pe-ripheral blood.GO and KEGG analysis of SSc DEGs showed that these genes were significantly enriched in inflammatory response,he-moglobin complex,immune receptor activity and extracellular matrix.The results of protein interaction network suggest that more than 10 genes such as COL1A1,CTGF12,IL1B,IFNG and JUN may be potential markers of SSc and core genes of therapeutic targets.The potential Chinese medicines screened for SSc treatment include ginseng,sanguisorba,convolvula,wolfberry,safflower,etc.The main components of these herbs were β-sitosterol,quercetin,kaempferol,stigmasterol,luteolin,sitosterol,Spinasterol,and the target were AKR1B1,AR,CYP1B1,XDH,etc.Conclusion:This study uses bioinformatics to screen out core genes that may be potential markers and therapeutic targets for SSc,which is expected to be a new target for the early diagnosis and mechanism research of SSc.Meanwhile,the mapped Chinese medicine and its effective components can provide ideas for the research and development of Chinese medicine compounds for the treatment of SSc.

Diabetes mellitus is the third most prevalent disease worldwide,following cardiovascular diseases and cancer.There is no obvious symptom in the early stage of diabetes mellitus.However,in the middle and late stages,diabetes mellitus may lead to severe clinical complications such as diabetic cardiomyopathy,kidney disease,retinopathy,or neuropathy,the primary causes for diabetes-related deaths.Therefore,the early diagnosis and treatment of diabetes mellitus are of practical significance.Compared with traditional diagnostic methods including fasting blood-glucose and oral glucose tolerance tests,the measurement of glycated hemoglobin serves as a gold standard for evaluating long-term blood glucose control,due to the relative stability of glycated hemoglobin under a period of dietary and daily practice.Ear-ly treatment of diabetes mellitus may significantly improve the prognosis of patients and enhance the overall therapeutic out-come.Diet control,physical exercise,medication,as well as psychological and social supports are critical for the treat-ment of diabetes mellitus.To date,the therapeutic methods for diabetes mellitus have been constantly enriching,along with promoted drug design and development.Some emerging technologies,such as therapies using antibodies or stem cells,have been applied to treat diabetes mellitus.Hereby the latest progress in the prevention and treatment of diabetes mellitus was comprehensively reviewed,and the application of antibodies was discussed,which may provide insights into the research and development of antibody drugs for chronic human diseases including diabetes mellitus.

Objective To explore the value of the ratio between inferior turbinate and nasal cavity morphology in the diagnosis and treatment of inferior turbinate hypertrophy.Methods The clinical and CT imaging data of patients with nasal septum deviation were retrospectively collected.Patients were divided into an inferior turbinate hypertrophy group(experimental group)and a control group without nasal congestion symptoms and related lesions.Based on CT imaging data,four ratios of inferior turbinate to nasal cavity morphology were calculated:inferior turbinate bone to nasal cavity width ratio,lateral mucosa of inferior turbinate to nasal cavity width ratio,medial mucosa of inferior turbinate to nasal cavity width ratio,and inferior turbinate to nasal cavity width ratio.A t-test was used to compare the four ratios between the experimental and control groups.Results The mean values of inferior turbinate bone to nasal cavity width ratio,lateral mucosa of inferior turbinate to nasal cavity width ratio,medial mucosa of inferior turbinate to nasal cavity width ratio,and inferior turbinate to nasal cavity width ratio at the uncinate process level in the experimental group were 0.344±0.046,0.101±0.031,0.108±0.031,and 0.244±0.043,respectively.In the control group,these ratios were 0.061±0.019,0.074±0.018,0.097±0.044,and 0.146±0.038,respectively.All four ratios were significantly higher in the experimental group than in the control group,with statistically significant differences(P＜0.05).The area under the curve(AUC)for diagnosing inferior turbinate hypertrophy using inferior turbinate bone to nasal cavity width ratio,lateral mucosa of inferior turbinate to nasal cavity width ratio,medial mucosa of inferior turbinate to nasal cavity width ratio,and inferior turbinate to nasal cavity width ratio were 0.870,0.907,0.884,and 0.926,respectively.Conclusion The ratios of inferior turbinate to nasal cavity morphology can serve as quantitative indicators for inferior turbinate hypertrophy,providing clinical value for treatment planning.

Objective To explore the value of the ratio between inferior turbinate and nasal cavity morphology in the diagnosis and treatment of inferior turbinate hypertrophy.Methods The clinical and CT imaging data of patients with nasal septum deviation were retrospectively collected.Patients were divided into an inferior turbinate hypertrophy group(experimental group)and a control group without nasal congestion symptoms and related lesions.Based on CT imaging data,four ratios of inferior turbinate to nasal cavity morphology were calculated:inferior turbinate bone to nasal cavity width ratio,lateral mucosa of inferior turbinate to nasal cavity width ratio,medial mucosa of inferior turbinate to nasal cavity width ratio,and inferior turbinate to nasal cavity width ratio.A t-test was used to compare the four ratios between the experimental and control groups.Results The mean values of inferior turbinate bone to nasal cavity width ratio,lateral mucosa of inferior turbinate to nasal cavity width ratio,medial mucosa of inferior turbinate to nasal cavity width ratio,and inferior turbinate to nasal cavity width ratio at the uncinate process level in the experimental group were 0.344±0.046,0.101±0.031,0.108±0.031,and 0.244±0.043,respectively.In the control group,these ratios were 0.061±0.019,0.074±0.018,0.097±0.044,and 0.146±0.038,respectively.All four ratios were significantly higher in the experimental group than in the control group,with statistically significant differences(P＜0.05).The area under the curve(AUC)for diagnosing inferior turbinate hypertrophy using inferior turbinate bone to nasal cavity width ratio,lateral mucosa of inferior turbinate to nasal cavity width ratio,medial mucosa of inferior turbinate to nasal cavity width ratio,and inferior turbinate to nasal cavity width ratio were 0.870,0.907,0.884,and 0.926,respectively.Conclusion The ratios of inferior turbinate to nasal cavity morphology can serve as quantitative indicators for inferior turbinate hypertrophy,providing clinical value for treatment planning.

Objective:The differentially expressed genes(DEGs)and activated signaling pathways in systemic sclerosis(SSc)were screened by bioinformatics methods,and Chinese medicines for potential treatment of SSc were explored,providing a new theoretical basis for the study of SSc and the screening of potential markers.Methods:The data sets GSE58095,GSE130953,GSE33463 and GSE58613 were selected from GEO database and divided into skin group and peripheral blood group according to the sample source.The DEGs of SSc patients was analyzed by R language,and the Wayne diagram was drawn to take the intersection of the two groups.Metascape was used for GO enrichment analysis and KEGG pathway enrichment analysis,and STRING and Cytoscape were used for protein interaction network analysis to find key pathways and hub genes.The core genes were mapped to the medical on-tology information retrieval platform,and related Chinese medicines for SSc treatment were screened.The effective components of Chi-nese medicines were obtained through TCMSP and HERB databases,and the target letters of active ingredients were obtained through swiss database.The"drug-active ingredient-target"network was constructed by Cytoscape.Results:Total 218 DEGs were identified from the skin group of patients with SSc,and 283 DEGs were screened from peripheral blood of patients with SSc.Among them,there were 7 DEGs co-upregulated in skin and peripheral blood,namely ISG15,LGALS3BP,BST2,C1QB,IFI27,CEACAM1 and FBP1.CAMK2N1 was up-regulated in skin but down-regulated in peripheral blood,ARG1 was down-regulated in skin but up-regulated in pe-ripheral blood.GO and KEGG analysis of SSc DEGs showed that these genes were significantly enriched in inflammatory response,he-moglobin complex,immune receptor activity and extracellular matrix.The results of protein interaction network suggest that more than 10 genes such as COL1A1,CTGF12,IL1B,IFNG and JUN may be potential markers of SSc and core genes of therapeutic targets.The potential Chinese medicines screened for SSc treatment include ginseng,sanguisorba,convolvula,wolfberry,safflower,etc.The main components of these herbs were β-sitosterol,quercetin,kaempferol,stigmasterol,luteolin,sitosterol,Spinasterol,and the target were AKR1B1,AR,CYP1B1,XDH,etc.Conclusion:This study uses bioinformatics to screen out core genes that may be potential markers and therapeutic targets for SSc,which is expected to be a new target for the early diagnosis and mechanism research of SSc.Meanwhile,the mapped Chinese medicine and its effective components can provide ideas for the research and development of Chinese medicine compounds for the treatment of SSc.

Diabetes mellitus is the third most prevalent disease worldwide,following cardiovascular diseases and cancer.There is no obvious symptom in the early stage of diabetes mellitus.However,in the middle and late stages,diabetes mellitus may lead to severe clinical complications such as diabetic cardiomyopathy,kidney disease,retinopathy,or neuropathy,the primary causes for diabetes-related deaths.Therefore,the early diagnosis and treatment of diabetes mellitus are of practical significance.Compared with traditional diagnostic methods including fasting blood-glucose and oral glucose tolerance tests,the measurement of glycated hemoglobin serves as a gold standard for evaluating long-term blood glucose control,due to the relative stability of glycated hemoglobin under a period of dietary and daily practice.Ear-ly treatment of diabetes mellitus may significantly improve the prognosis of patients and enhance the overall therapeutic out-come.Diet control,physical exercise,medication,as well as psychological and social supports are critical for the treat-ment of diabetes mellitus.To date,the therapeutic methods for diabetes mellitus have been constantly enriching,along with promoted drug design and development.Some emerging technologies,such as therapies using antibodies or stem cells,have been applied to treat diabetes mellitus.Hereby the latest progress in the prevention and treatment of diabetes mellitus was comprehensively reviewed,and the application of antibodies was discussed,which may provide insights into the research and development of antibody drugs for chronic human diseases including diabetes mellitus.

General anesthesia plays a significant role in modern medicine. However, the precise mechanism of general anesthesia remains unclear, posing a key scientific challenge in anesthesiology. Advances in neuroscience techniques have enabled targeted manipulation of specific neural circuits and the capture of brain-wide neural activity at high resolution. These advances hold promise for elucidating the intricate mechanisms of action of general anesthetics. This review aims to summarize our current understanding of the role of cortical and subcortical nuclei in modulating general anesthesia, providing new evidence of cortico-cortical and thalamocortical networks in relation to anesthesia and consciousness. These insights contribute to a comprehensive understanding of the neural network mechanisms underlying general anesthesia.
Humans ; Anesthesia, General ; Animals ; Nerve Net/physiology* ; Cerebral Cortex/drug effects* ; Neural Pathways/drug effects* ; Thalamus/drug effects* ; Consciousness/drug effects*

Rheumatoid arthritis (RA) is an autoimmune disease characterized by severe synovial inflammation and cartilage damage. Despite great progress in RA therapy, there still lacks the drugs to completely cure RA patients. Herein, we propose a reprogrammed neutrophil cytopharmaceuticals loading with TNFα-targeting-siRNA (siTNFα) as an alternative anti-inflammatory approach for RA treatment. The loaded siTNFα act as not only the gene therapeutics to inhibit TNFα production by macrophages in inflamed synovium, but also the editors to reprogram neutrophils to anti-inflammatory phenotypes. Leveraging the active tendency of neutrophils to inflammation, the reprogrammed siTNFα/neutrophil cytopharmaceuticals (siTNFα/TP/NEs) can rapidly migrate to the inflamed synovium, transfer the loaded siTNFα to macrophages followed by the significant reduction of TNFα expression, and circumvent the pro-inflammatory activity of neutrophils, thus leading to the alleviated synovial inflammation and improved cartilage protection. Our work provides a promising cytopharmaceutical for RA treatment, and puts forward a living neutrophil-based gene delivery platform.