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 Following four immunization sessions, blood was withdrawn from the jugular vein, and a nanobody library with a capacity of 2.85×10⁸ colony forming units (CFU) was generated.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 evaluate the changes in cerebral perfusion after moyamoya disease revascularization using two-phase arterial spin labeling(ASL)combined with territorial arterial spin labeling(t-ASL).Methods A retrospective selection was conducted on 34 moy-amoya disease patients with 37 cerebral hemispheres who underwent revascularization surgery.ASL[post labeling delay(PLD)of 1.5 s and 2.5 s],t-ASL,and digital subtraction angiography(DSA)were performed before and after surgery.The cerebral blood flow(CBF)and relative cerebral blood flow(rCBF)values in the middle cerebral artery(MCA)supply area from ASL were com-pared before and after surgery.t-ASL and DSA were used to score the area of revascularization on a 3-point scale,and weighted Kappa was used to compare the consistency of the revascularization area assessments.A DSA score greater than 1 point was classified as good revascularization.The diagnostic efficacy of ASL(PLD 1.5 s,2.5 s)and t-ASL for revascularization were compared.Results In ASL PLD 1.5 s,the preoperative and postoperative CBF values were(32.65±12.71)mL·100 g-1·min-1 and(43.09±18.05)mL·100 g-1·min-1,respectively,and the rCBF values were 0.76±0.23 and 1.02±0.31;In ASL PLD 2.5 s,the preoperative and postoperative CBF val-ues were(47.07±13.28)mL·100 g-1·min-1and(52.85±14.98)mL·100 g-1·min-1,respectively,and the rCBF values were 1.21±0.31 and 1.33±0.29.There were statistically significant differences between preoperative values and postoperative values above(P<0.05).The revascularization area score showed strong consistency(weighted Kappa coefficient=0.729).Receiver operating charac-teristic(ROC)curve analysis revealed that t-ASL reflected the revascularization area with area under the curve(AUC)of 0.982,while ASL PLD 1.5 s and 2.5 s CBF reflected the revascularization area with AUC of 0.813 and 0.794,espectively.Conclusion The com-bination of two-phase ASL and t-ASL more effectively evaluates cerebral blood after moyamoya disease revascularization.Furthermore,t-ASL demonstrates higher efficacy in evaluating postoperative revascularization.

Objective To evaluate the changes in cerebral perfusion after moyamoya disease revascularization using two-phase arterial spin labeling(ASL)combined with territorial arterial spin labeling(t-ASL).Methods A retrospective selection was conducted on 34 moy-amoya disease patients with 37 cerebral hemispheres who underwent revascularization surgery.ASL[post labeling delay(PLD)of 1.5 s and 2.5 s],t-ASL,and digital subtraction angiography(DSA)were performed before and after surgery.The cerebral blood flow(CBF)and relative cerebral blood flow(rCBF)values in the middle cerebral artery(MCA)supply area from ASL were com-pared before and after surgery.t-ASL and DSA were used to score the area of revascularization on a 3-point scale,and weighted Kappa was used to compare the consistency of the revascularization area assessments.A DSA score greater than 1 point was classified as good revascularization.The diagnostic efficacy of ASL(PLD 1.5 s,2.5 s)and t-ASL for revascularization were compared.Results In ASL PLD 1.5 s,the preoperative and postoperative CBF values were(32.65±12.71)mL·100 g-1·min-1 and(43.09±18.05)mL·100 g-1·min-1,respectively,and the rCBF values were 0.76±0.23 and 1.02±0.31;In ASL PLD 2.5 s,the preoperative and postoperative CBF val-ues were(47.07±13.28)mL·100 g-1·min-1and(52.85±14.98)mL·100 g-1·min-1,respectively,and the rCBF values were 1.21±0.31 and 1.33±0.29.There were statistically significant differences between preoperative values and postoperative values above(P<0.05).The revascularization area score showed strong consistency(weighted Kappa coefficient=0.729).Receiver operating charac-teristic(ROC)curve analysis revealed that t-ASL reflected the revascularization area with area under the curve(AUC)of 0.982,while ASL PLD 1.5 s and 2.5 s CBF reflected the revascularization area with AUC of 0.813 and 0.794,espectively.Conclusion The com-bination of two-phase ASL and t-ASL more effectively evaluates cerebral blood after moyamoya disease revascularization.Furthermore,t-ASL demonstrates higher efficacy in evaluating postoperative revascularization.

ObjectiveTo investigate the therapeutic effect of Qinggongtang in regulating Glu/GABA metabolic balance and the mechanism of its anxiolytic effect on rat models of anxiety. MethodFifty-four rats were randomly divided into normal， model， diazepam （0.225 mg·kg^-1）， and low-dose， medium-dose， and high-dose groups of Qinggongtang （5.085， 10.17， 20.34 g·kg^-1）， with nine rats in each group. Except for the normal group， the other groups were subjected to indeterminate vacutainer stress and chronic restraint stress for 12 days to prepare the anxiety model. On the 3^rd day of the stress， 10 days of corresponding drug intervention was started. At the end of the drug treatment， the anxiety level of rats in each group was evaluated by the elevated cross maze experiment （EPM） and the light and dark box experiment （LDB）， and the effect of Qinggongtang on the anxiety behavior of rats was preliminarily analyzed. The levels of Glu and GABA in the amygdala tissue of the rats were detected by enzyme linked immunosorbent assay （ELISA）， and the changes in the synaptic ultrastructure of the amygdala of the rats in each group were observed by electron microscopy. The mRNA expression of glutamic acid decarboxylase （GAD65 and GAD67）， glutamine synthetase （GS）， and glutamate transporter-1 （GLT-1） in the amygdala were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， and their protein expression was detected by Western blot. ResultCompared with those in the normal group， rats in the model group showed an obvious anxiety state and dull yellow and lusterless fur. They were irritable， easy to anger， and preferred to curl up in the corner. The number of times the EPM entered the open arm and the residence time in the open arm were significantly reduced （P<0.01）， and the residence time in the open box and the number of times the LDB went through the box were significantly reduced （P<0.01）. The content of Glu in the amygdala was increased （P<0.01）， and the content of GABA was reduced （P<0.01）. The value of Glu/GAB was elevated （P<0.01）， and the number of synaptic and pre-synaptic membrane vesicles in the amygdala was decreased. Sparse dense material in the post-synaptic membrane， increased synaptic gap， slightly disrupted internal structure， and decreased mRNA and protein expressions of GAD65， GAD67， GS， and GLT-1 in the amygdala were observed （P<0.01）. Compared with those in the model group， rats in the medium-dose and high-dose groups of Qinggongtang and the diazepam group had bright fur， sensitive reactions， and more active behavior. The number of times EPM entered the open arm and the residence time in the open arm increased significantly （P<0.01）， and the residence time in the open box and the number of times the LDB went through the box increased significantly （P<0.01）. The content of Glu in all-dose groups of Qinggongtang and the diazepam group decreased （P<0.05， P<0.01）， while GABA content increased （P<0.05， P<0.01）. The value of Glu/GABA decreased （P<0.01）， and the internal and external synaptic structure of each groups of Qinggongtang and the diazepam group was more complete. Synapses and vesicles were numerous， and the synaptic gap was more clearly defined. The efficacy of the high-dose group of Qinggongtang and the diazepam group was the best， and the mRNA and protein expressions of GAD65， GAD67， GS， and GLT-1 in the amygdala were increased in the high-dose group of Qinggongtang and diazepam group （P<0.05， P<0.01）. ConclusionQinggongtang can improve synaptic plasticity and affect the expression of GAD65， GAD67， GS， and GLT-1 in the amygdala of rats to regulate Glu/GABA metabolic balance and thus exert anxiolytic effects.

Somatic mutations in the promoter region of telomerase reverse transcriptase(TERT),a critical mechanism for telomerase reactivation,play a key role in tumorigenesis.The status of TERT promoter mutation serves as a crucial molecular biomarker for glioma assessment and classification,and is essential for early diagnosis of glioma subtypes,guiding treatment decision-making,and improving patient prognosis.With the recognition of the importance of molecular subtyping of gliomas,there has been a surge in research on non-invasive prediction of key molecular biomarkers based on preoperative imaging of gliomas,with a particular focus on TERT promoter studies using radiomics approaches.This article presents a comprehensive review of research on TERT promoter mutations in gliomas and imaging-related studies,with the goal of providing insights for future studies on non-invasive prediction of TERT promoters status and offering important references for the precision diagnosis and treatment of glioma patients.

Objective:To study the short-term clinical outcomes of different courses of antenatal corticosteroids (ACS) for preterm twins.Methods:From January 2017 to December 2021, preterm twins with gestational age (GA) 24-34 weeks admitted to the neonatal ward of our hospital and received ACS were retrospectively studied. The infants were assigned into single-course group, partial-course group and multiple-course group according to ACS courses. The short-term clinical outcomes were compared among the groups. SPSS software version 25.0 was used for statistical analysis.Results:A total of 286 infants were enrolled in this study, including 128 in single-course group, 89 in partial-course group and 69 in multiple-course group. Compared with single-course group, the risks of neonatal respiratory distress syndrome (RDS) in both partial-course group ( OR=2.332, 95% CI 1.028-5.293, P=0.043) and multiple-course group ( OR=3.872, 95% CI 1.104-13.584, P=0.034) were higher. The birth length in multiple-course group ( β=-0.016, 95% CI -0.029 - -0.002, P=0.024) was lower than single-course group. Conclusions:The risks of neonatal RDS in preterm twins are higher in partial-course and multiple-course of ACS. A full course of ACS should be used to prevent neonatal RDS until further evidence of effectiveness is available.

The use of bariatric and metabolic surgery as a central treatment for obesity has been steadily increasing. BMI, as a widely used metric for assessing obesity, has considerable relevance in the field of metabolic research. However, its limitations, such as its inability to account for variations in fat distribution, remain a subject of considerable controversy. In recent years, there has been a surge of interest in the relationship between changes in body composition and the risk of metabolic disease. Consequently, the study of the effects of bariatric and metabolic surgery on changes in body composition has become a major focus of bariatric and metabolic surgery research. As a potential replacement for BMI, body composition measurements are expected to improve and standardize the assessment of the effectiveness of bariatric and metabolic surgery. This underscores the urgent need for the development of methods and standards for body composition measurement. This paper undertakes a comprehensive review of the existing evidence on the application of body composition measurement techniques for the efficacy evaluation of bariatric and metabolic surgery. The intent is to provide new insights and pave the way for the exploration of future research directions in this area.

The use of bariatric and metabolic surgery as a central treatment for obesity has been steadily increasing. BMI, as a widely used metric for assessing obesity, has considerable relevance in the field of metabolic research. However, its limitations, such as its inability to account for variations in fat distribution, remain a subject of considerable controversy. In recent years, there has been a surge of interest in the relationship between changes in body composition and the risk of metabolic disease. Consequently, the study of the effects of bariatric and metabolic surgery on changes in body composition has become a major focus of bariatric and metabolic surgery research. As a potential replacement for BMI, body composition measurements are expected to improve and standardize the assessment of the effectiveness of bariatric and metabolic surgery. This underscores the urgent need for the development of methods and standards for body composition measurement. This paper undertakes a comprehensive review of the existing evidence on the application of body composition measurement techniques for the efficacy evaluation of bariatric and metabolic surgery. The intent is to provide new insights and pave the way for the exploration of future research directions in this area.

ObjectiveTo observe the intervention effect of Ruyi Zhenbao pills （RYZBP） on central pain after thalamic stroke in mice and explore the underlying mechanism. MethodThe central post-stroke pain syndrome （CPSP） model was induced by stereotactic injection of type Ⅳ collagenase into the hypothalamus in mice. The mice were divided into a sham group， a model group， low-， medium-， and high-dose RYZBP groups （0.65， 1.3， 2.6 g·kg^-1）， and a pregabalin group （0.075 g·kg^-1）. Seven days after modeling， the mice in the groups with drug intervention were administered with corresponding drugs by gavage according to the body mass， once per day for 25 days， while those in the sham group and the model group received an equal volume of normal saline. During this period， mechanical pain and cold pain were detected at different time points， and the apoptotic state of brain tissue cells was detected by in situ terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling （TUNEL）. The 36 classical broad-spectrum inflammatory factors were quantitatively analyzed by liquid-phase chip technology， and differential molecules were screened out and verified by Western blot and enzyme-linked immunosorbent assay （ELISA）. ResultCompared with sham operation group, mechanical pain threshold and cold sensitive pain threshold in model group were significantly changed （P<0.01）. TUNEL results showed that apoptosis of brain cells was obvious. Western blot and ELISA results showed that the expressions of interleukin-1α (IL-1α) and chemokine ligand 5 (CCL5) increased in hypothalamus tissue and serum， while the expressions of Ang-2, granulocyte-colony-stimulating factor (G-CSF) and IL-4 decreased significantly （P<0.01）. Compared with model group， RYZBW dose groups significantly increased mechanical pain threshold， decreased cold sensitivity pain threshold， decreased hypothalamus cell apoptosis ratio （P<0.01）， decreased the expression of IL-1α and CCL5 in hypothalamus tissue and serum， while the expression of ANG-2， G-CSF and IL-4 were significantly increased （P<0.05）. ConclusionRYZBP can relieve hyperalgesia in CPSP mice， and its mechanism is related to the regulation of the expression of pro-/anti-inflammatory factors IL-1α， CCL5， IL-4， G-CSF， and Ang-2.

BACKGROUND: Perioperative treatment has become an increasingly important aspect of the management of patients with non-small cell lung cancer (NSCLC). Small-scale clinical studies performed in recent years have shown improvements in the major pathological remission rate after neoadjuvant therapy, suggesting that it will soon become an important part of NSCLC treatment. Nevertheless, neoadjuvant immunotherapy may be accompanied by serious adverse reactions that lead to delay or cancelation of surgery, additional illness, and even death, and have therefore attracted much attention. The purpose of the clinical recommendations is to form a diagnosis and treatment plan suitable for the current domestic medical situation for the immune-related adverse event (irAE). METHODS: This recommendation is composed of experts in thoracic surgery, oncologists, thoracic medicine and irAE related departments (gastroenterology, respirology, cardiology, infectious medicine, hematology, endocrinology, rheumatology, neurology, dermatology, emergency section) to jointly complete the formulation. Experts make full reference to the irAE guidelines, large-scale clinical research data published by thoracic surgery, and the clinical experience of domestic doctors and publicly published cases, and repeated discussions in multiple disciplines to form this recommendation for perioperative irAE. RESULTS: This clinical recommendation covers the whole process of prevention, evaluation, examination, treatment and monitoring related to irAE, so as to guide the clinical work comprehensively and effectively. CONCLUSIONS Perioperative irAE management is an important part of immune perioperative treatment of lung cancer. With the continuous development of immune perioperative treatment, more research is needed in the future to optimize the diagnosis and treatment of perioperative irAE.