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 Following four immunization sessions, blood was withdrawn from the jugular vein, and a nanobody library with a capacity of 2.85×10⁸ colony forming units (CFU) was generated.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 factors influencing the pregnancy outcomes during frozen-thawed embryo transfer(FET)cycles in patients with polycystic ovary syndrome(PCOS).Methods A retrospective analysis was conducted on patients'data from 882 FET cycles.According to the pregnancy outcome,the patients were divided into non-implantation group(Group A),abortion group(Group B1)and live birth group(Group B2).Clinical data and laboratory parameters were compared among the three groups,and ordered Logistic regression analysis was used to study the factors influencing pregnancy outcomes after FET.Patients were also divided into four groups(C1-C4)based on the number of high-quality embryos obtained(0-3,4-6,7-10,≥11),and their clinical data and laboratory parameters were compared.Results The clinical pregnancy rate,live birth rate,and miscar-riage rate in the 882 treatment cycles were 71.09%(627/882),61.68%(544/882),and 13.24%(83/627),respectively.Single-factor analysis showed significant differences in body mass index(BMI),infertility type,hu-man chorionic gonadotropin(hCG)day estradiol(E2)level,number of retrieved oocytes,and number of high-quality embryos among Groups A,B1,and B2(P<0.05).Further multiple Logistic regression analysis revealed that BMI(OR=1.046,95%CI:1.001-1.093,P=0.044)and a history of previous pregnancy(OR=1.417,95%CI:1.030-1.950,P=0.032)were independent risk factors for successful FET in PCOS patients,while an in-creased number of high-quality embryos was an independent protective factor for successful pregnancy.Based on the results of Group B2,compared to Group A,OR=0.920,95%CI:0.880-0.962,P=0.000;compared to Group B1,OR=0.923,95%CI:0.862-0.988,P=0.022.Compared with the other three groups(C1-C3),the total amount of gonadotropin(Gn)in the C4 group was the lowest and the number of oocytes obtained was the high-est(P<0.05).Multiple comparisons showed that Group C4 had lower BMI,follicle-stimulating hormone(FSH),very low-density lipoprotein(vLDL)levels,a higher luteinizing hormone and follicle-stimulating hormone(LH/FSH)ratio compared to Group C1(P<0.05).Group C4 had lower fasting insulin(FINS)and homeostasis model assessment of insulin resistance(HOMA-IR)levels compared to Group C3,and higher high-density lipoprotein-cholesterol(HDL-C)and apolipoprotein A1(Apo A1)levels compared to Groups C2 and C3(P<0.05).Con-clusion BMI,the history of previous pregnancy and the number of high-quality embryos were both independent factors for predicting pregnancy outcomes in PCOS patients undergoing FET cycles.Patients with a higher number of high-quality embryos have a higher clinical pregnancy rate during FET cycles.

Objective:To investigate the effect of combined use of midazolam and remifentanil without muscle relaxant on tracheal intubation in female patients undergoing breast surgery.Methods:A retrospective analysis was performed on 40 female patients with breast disease who underwent tracheal intubation and general anesthesia at The First Affiliated Hospital of Zhengzhou University between January 2023 and June 2023. These patients were divided into a control group ( n = 20) and an observation group ( n = 20) based on whether muscle relaxants were applied at the time of intubation. The control group received intravenous rocuronium bromide, whereas the observation group did not use muscle relaxants. Both groups were intravenously administered midazolam (0.1 mg/kg) and remifentanil (4 μg/kg) prior to tracheal intubation. The intubation conditions were evaluated based on factors such as the ease of inserting the laryngoscope and the patient's response to intubation, including coughing. Results:There were no statistically significant differences in age, height, and body mass between the two groups (all P > 0.05). The excellent rate of intubation conditions was significantly lower in the observation group compared with the control group [45% (9/20) vs. 85% (17/20), χ2 = 7.03, P = 0.008). The good rate of intubation conditions was significantly higher in the observation group compared with the control group [40% (8/20) vs. 5% (1/20), χ2 = 7.03, P < 0.05]. There was no statistically significant difference in the excellent and good rates of intubation conditions between the observation and control groups [85% (17/20) vs. 90% (18/20), χ2 = 0.23, P > 0.05]. No significant difference in intraoperative awareness score was observed between the observation and control groups [(2.59 ± 0.44) points vs. (2.61 ± 0.31) points, P > 0.05]. None of the patients in either group exhibited any episodes of arrhythmias. Furthermore, no adverse reactions such as muscle stiffness, nausea, vomiting, or skin itching were observed in either group following the surgical procedure. Conclusion:Without the use of muscle relaxants, intravenous administration of midazolam at 0.1 mg/kg and remifentanil at 4 μg/kg for tracheal intubation in female patients undergoing breast surgery can offer excellent intubation conditions, ensuring that the patient remains unconscious throughout the surgical procedure.

OBJECTIVE To identify an d analyze chemical cons tituents of Pleione yunnanensis with origin of Pleione yunnanensis. METHODS UPLC-Q-Exactive-Plus-Orbitrap-MS was adopted. The determination was performed on Hyperdil GOLD column with mobile phase consisted of 0.1％ formic acid solution-0.1％ formic acid acetonitrile solution （gradient elution ）at the flow rate of 0.3 mL/min. The column temperature was set at 40 ℃，and sample size was 2 µL. The electrospray ion source was adopted，and the scanning range was m/z 100-1 500，and the scanning mode was positive and negative ion exchange mode of full scan+ddMS2. The structure of chemical constituents were determined by using Compound Discoverer 3.1 software，comparing with mzCloud，PubChem network database and OTCML ，on the basis of reference substance and published literatures. RESULTS & CONCLUSIONS A total of 42 chemical constituents were identified （positive ion mode has 24，negative ion mode has 27）， including 13 benzyl succinate glycosides （such as dactylorhin C ，coelovirin A ，militarine），4 phenol glycosides （such as adenosine ， guanosine，gastrodin），3 alkaloids（choline，betaine，berberine），and one flavonoid （nobiletin），7 aromatics（such as DL-lysine ， DL-arginine，DL-glutamine），one sugar （sucrose），3 benzenes（shancigusin H ，shancigusin H isomer ，batatasin Ⅲ）and 10 others （such as p-methoxybenzoic acid ，monomethyl dodecanedioate ，diphenylamine）. Glucose oxybenzyl and some small mole cules are easy to be lost in the cleavage of benzyl succinate glycosides；glycosyl is easy to be lost in the cleavage of phenol glycosides；the cleavage of alkaloids mainly manifest as the cleavage and loss of small molecular substituents ；demethyla- tion reaction is occurred in most flavonoids.

Objective To determine the occurrence and clinical characteristics of ectopic pituitary adenoma (EPA) in China.Methods The study are done by searching systematically and comprehensively on major Chinese and English medical literature databases and conference papers before 2015,which are only limited to collected,summarized,sorted and analyzed EPA cases that reported by Chinese authors in English or Chinese occurred in China.Results ① Among the 86 Chinese articles and 27 English articles related to ectopic pituitary adenoma (EPA) gathered by the author,except for cases that have been confirmed as repeated reports,a total of 73 EPA cases were found.Of 70 cases with complete data,31 were male cases,accounting for 44.29％;39 were female cases,accounting for 55.71％;the ratio is 1:1.3.The frequency of EPA occurrence according to the location of the lesion,from high to low in turn,was sphenoid sinus (31 cases,42.47％),nasopharynx (7 cases,9.59％),suprasellar region (7 cases,9.59％),clivus (5 cases,6.85％),followed by the third ventricle,sphenoid sinus/clivus,nasal cavity,and the temporal lobe of the brain,with the same incidence of 4.11％.According to the functional properties of EPA,the frequency of different secreting hormones adenoma was PRL-ma(21 cases,28.77％),NF-ma (21 cases,28.77％),ACTH-ma (15 cases,20.55％),GH-ma (10 cases,13.70％),TSH-ma (2 cases,2.74％) and FSH-ma (1 cases.1.37％).Three cases of EPA were uncertain in their property due to lack of information.The incidence of PRL and nonfunctional tumors was the highest,which was different from what was reported in other countries.Among them,one case of EPA was in pineal region and one in parapharyngeal space,which was even more rare and were never reported abroad.(② Except for 3 cases with incomplete medical records,15 out of 70 cases of EPA were accompanied by empty sella,accounting for 21.43％,among which 11 (73.33％) cases involved the sphenoid sinus,and 3 (20％) cases involved clivus.The sphenoid sinus and clivus cases together accounted for 93.33％.(③ 29 out of the 69 cases of EPA with complete record were invasive pituitary adenomas,accounting for 42.03％ and including 1 case of pituitary adenocarcinoma,which accounted for 1.45％.(④ All cases were treated with surgery as the first choice,and some of them were treated with radiotherapy or drug therapy.Conc lusion Ectopic pituitary adenoma is extremely rare.By the end of 2015,the total number of cases reported in China is only 73,which are mostly located in the sphenoid sinus,suprasellar region and nasopharynx.In the functional classification,the most common types are still PRL adenoma and nonfunctional adenoma as in situ pituitary adenoma.42.03％ of EPA adenomas are invasive.Surgical resection of EPA is the first choice and some cases can be treated with radiotherapy and drug therapy.

Objective To investigate the effect of health education for discharged patients, and to explore the reasons and effective countermeasures on the health education. Methods Through the question-naire survey, convenient sampling method was used to evaluate the effect of the hospital health education. The existing problems and corresponding countermeasures taken for continuous improvement were analyzed. SPSS 17.0 statistical software was applied for data processing, the count data was expressed by rate and Statistical methods using chi-square test. Results The hospital health education satisfaction rate rose to 95.21%from 54.17%after intervention, and differences between the two groups was statistically significant (P<0.05). The satisfaction rate of the department of internal medicine, surgery department, and the department of gynaecology and pediatrics was significantly improved than before. Conclusion By improving health education importance, strengthening learning and training of specialized subject knowledge, rationally allo-cating human resources and establishing health education quality control system, the health education for discharged patients was effectively improved with satisfaction.