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 Following four immunization sessions, blood was withdrawn from the jugular vein, and a nanobody library with a capacity of 2.85×10⁸ colony forming units (CFU) was generated.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 explore the efficacy of low-frequency repetitive transcranial magnetic stimulation(rTMS)combined with virtual reality interactive robot training in improving upper limb function of patients with stroke.Methods From February to December 2023,92 patients in the hos-pital were randomly divided into control group(n=30),virtual reality group(n=31),and com-bined group(n=31).The control group received conventional rehabilitation therapy;the virtual re-ality group received conventional rehabilitation therapy and virtual reality interactive robot training for upper limb;the combined group received low-frequency rTMS on the therapeutic basis of the virtual reality group.Before treatment and 4 weeks after treatment,the Upper Extremity Fugl-Meyer Assess-ment(UFMA)score,the Functional Test for the Hemiplegic Upper Extremity-Hong Kong Version(FTHUE-HK)score,motor evoked potential(MEP)amplitude,cortical latency(CL)value,and the ratio of root mean square of myoelectricity(RMS)of wrist dorsiflexor muscles between the affect-ed and unaffected sides were compared among the three groups.Results Four weeks after treatment,the UFMA and FTHUE-HK scores of the three groups significantly improved compared with those before treatment,the UFMA and FTHUE-HK scores of the combined group were significantly higher than those of the control group and the virtual reality group,and the UFMA score of the virtual reality group was significantly higher than that of the control group(P＜0.05);the RMS ratios and MEP amplitudes of the three groups significantly increased compared with those before treatment,the RMS ratios and MEP amplitudes of the combined group were significantly higher than those of the control group and the virtual reality group,and the virtual reality group had higher values than the control group,with significant between-group differences(P＜0.05);the CL of the three groups significantly shortened compared with that before treatment,the CL of the combined group was significantly shorter than that of the control group and the virtual reality group,and the CL of the virtual reality group was significantly shorter than that of the control group(P＜0.05).Conclusion The rTMS assisted virtu-al reality interactive robot training can effectively improve upper limb function in stroke patients.

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.

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.

Pheochromocytomas and paragangliomas are rare but therapeutically challenging neuroendocrine tumors in urology. Surgical treatment has been recognized as a definitive treatment. However, traditional surgical methods present certain risks and limitations. The Da Vinci robotic surgery offers a new approach for treating these tumors. This review elucidates the technical features, clinical applications, and treatment outcomes of Da Vinci robotic surgery and the progress in the treatment of pheochromocytomas and paragangliomas. In addition, this review discusses the prospects of combining Da Vinci robotic surgery with other emerging technologies, emphasizing the further research to validate its long-term efficacy and safety.

Objective To explore the effect of task-oriented training of intelligent mirror gloves combined with low-frequency repeti-tive transcranial magnetic stimulation(rTMS)on hand function recovery in stroke patients. Methods From October 1st,2022 to June 30th,2023,136 stroke patients in Northern Jiangsu People's Hospital were ran-domly divided into control group,mirror group,rTMS group and combination group,with 34 patients in each group.All the groups received routine rehabilitation treatment.In addition,the mirror group received task-orient-ed training of intelligent mirror gloves,rTMS group received low-frequency rTMS,and the combination group received task-oriented training combined with low-frequency rTMS,for four weeks.The Fugl-Meyer Assess-ment-Upper Extremities(FMA-UE)score,Wolf Motor Function Test(WMFT)score,and surface electromyo-graphic root mean square(RMS)of forearm extensor and flexor muscle groups on the affected/healthy side be-fore and after treatment were compared.And the differences of transcranial magnetic stimulation-motor-evoked potentials(MEP)between rTMS group and combination group before and after treatment were also compared. Results Four cases in the control group,seven in the mirror group,five in rTMS group and six in the combination group dropped off.The intra-group effect(F>996.656,P<0.001),inter-group effect(F>20.333,P<0.001)and inter-action effect(F>72.796,P<0.001)were significant in the scores of FMA-UE and WMFT,and the RMS ratio of forearm extensor and flexor muscle groups among four groups,in which the combination group was the best.After treatment,the amplitude of MEP increased in rTMS group and combination group(|t|>3.842,P<0.05),and was higher in the combination group than in rTMS group(t=-3.060,P<0.01). Conclusion The task-oriented training of intelligent mirror gloves combined with low-frequency rTMS could effectively promote the recovery of hand function in stroke patients.

Pheochromocytomas and paragangliomas are rare but therapeutically challenging neuroendocrine tumors in urology. Surgical treatment has been recognized as a definitive treatment. However, traditional surgical methods present certain risks and limitations. The Da Vinci robotic surgery offers a new approach for treating these tumors. This review elucidates the technical features, clinical applications, and treatment outcomes of Da Vinci robotic surgery and the progress in the treatment of pheochromocytomas and paragangliomas. In addition, this review discusses the prospects of combining Da Vinci robotic surgery with other emerging technologies, emphasizing the further research to validate its long-term efficacy and safety.

Stroke often causes severe motor, sensory, and daily living function impairments, especially the recovery of distal limb extensor motor function is the most difficult. With the widespread application of Contralateral Control Functional Electrical Stimulation (CCFES) in stroke rehabilitation and continuous improvement of integrated wearable devices in recent years, it has been found that CCFES and combination therapy have good therapeutic effects in improving wrist extension and ankle dorsiflexion function in stroke patients. CCFES can improve both distal and proximal upper limb function, when applied to lower limbs, attention should be paid to the reverse coordination mechanism. Early intervention, sufficient treatment courses, and multiple combination CCFES treatment plans can accelerate the improvement of stroke patients' function.

Objective:To investigate the relationship between enteral nutrition-related diarrhea and intestinal flora in critically ill patients and the effect of microflora transplantation.Methods:A total of 60 critically ill patients with enteral nutrition-related diarrhea who were scheduled to undergo microflora transplantation in Taizhou Hospital of Integrated Traditional Chinese and Western Medicine from January 2020 to August 2021 were prospectively and continuously selected as the research group, and 60 critically ill patients without enteral nutrition-related diarrhea were selected as the control group. The bacterial count of 4 kinds of intestinal flora in the feces including bifidobacterium, lactobacillus, enterococcus, and escherichia coli were detected and compared between the two groups, and the value of the fecal colony numbers of 4 kinds of intestinal flora in diagnosing non-enteral nutrition-related diarrhea in critically ill patients was analyzed by receiver operating characteristic (ROC) curve. All patients in the research group received microflora transplantation, and the diarrhea score, hematochezia score, partial Mayo score and European five-dimension health scale (EQ-5D) were detected and compared before treatment, 1 week after treatment and 1 month after treatment to evaluate the treatment effect. The Pearson linear correlation method was used to analyze the relationship between the colony count of 4 kinds of intestinal flora colonies in the feces of the research group at baseline and the therapeutic indexes for 1 week and 1 month after treatment.Results:The number of fecal bifidobacterium and lactobacillus colonies in the study group were lower than those in the control group: (7.12 ± 0.58) × 10 7 cfu/L vs. (11.85 ± 1.25) × 10 7 cfu/L, (8.78 ± 1.05) × 10 7 cfu/L vs. (11.25 ± 1.57) ×10 7 cfu/L. The colony number of enterococcus and Escherichia coli were higher than those of control group: (8.58 ± 0.88) × 10 7 cfu/L vs. (3.84 ± 0.72) ×10 7 cfu/L, (8.25 ± 0.97) ×10 7 cfu/L vs. (3.66 ± 0.63) ×10 7 cfu/L. The differences were statistically significant ( P<0.05). ROC curve analysis results showed that the area under the curve of fecal bifidobacterium, lactobacillus, enterococcus and escherichia coli colonies in diagnosing patients with enteral nutrition-related diarrhea were all >0.7, which had certain diagnostic value. The diarrhea scores, stool blood scores and some Mayo scores of the study group at 1 week and 1 month after treatment were lower than those before treatment: (1.52 ± 0.36) and (1.13 ± 0.24) points vs. (2.45 ± 0.51) points, (0.95 ± 0.28) and (0.77 ± 0.21) points vs. (2.39 ± 0.54) points, (4.17 ± 1.24) and (3.26 ± 0.85) points vs. (7.86 ± 1.82) points, and the EQ-5D score of patients 1 week and 1 month after treatment was higher than that before treatment: (0.66 ± 0.11) and (0.79 ± 0.13) points vs. (0.58 ± 0.08) points, the difference was statistically significant ( P<0.05). Conclusions:The intestinal flora of critically ill patients is closely related to enteral nutrition-related diarrhea, and can affect the therapeutic effect of bacterial flora transplantation and the health status of patients.

Coronavirus disease (COVID-19) is an infectious disease caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), with strong contagiousness, high susceptibility and long incubation period. cell entry by SARS-CoV-2 requires the binding between the receptor-binding domain of the viral spike protein and the cellular angiotensin-converting enzyme 2 (ACE2). Here, we briefly reviewed the mechanisms underlying the interaction between SARS-CoV-2 and ACE2, and summarized the latest research progress on SARS-CoV-2 neutralizing monoclonal antibodies and nanobodies, so as to better understand the development process and drug research direction of COVID-19. This review may facilitate understanding the development of neutralizing antibody drugs for emerging infectious diseases, especially for COVID-19.
Angiotensin-Converting Enzyme 2 ; Antibodies, Monoclonal ; Antibodies, Neutralizing ; Antibodies, Viral ; COVID-19 ; Humans ; Peptidyl-Dipeptidase A/metabolism* ; Protein Binding ; SARS-CoV-2 ; Single-Domain Antibodies ; Spike Glycoprotein, Coronavirus/metabolism*