Objective The study aims to investigate the immunological functions of the nucleocapsid (N) protein of the novel coronavirus Omicron (BA.1, BA.2) and evaluate the differences among different N proteins of mutant strains in immunogenicity. Methods By aligning sequences, the mutation sites of the Omicron (BA.1, BA.2) N protein relative to prototype strain of the novel coronavirus (Wuhan-Hu-1) were determined. The pET-28a-N-Wuhan-Hu-1 plasmid was used as template to construct pET-28a-BA.1/BA.2-N through single point mutation or homologous recombination. The three kinds of N protein were expressed in prokaryotic system, purified through Ni-NTA affinity chromatography, and then immunized into mice. The titer and reactivity of the polyclonal antibody, as well as the expression level of IL-1β and IFN-γ in mouse spleen cells, were detected using indirect ELISA and Western blot assay. Results The constructed prokaryotic expression plasmids were successfully used to express the Wuhan-Hu-1 N, BA.1 N, and BA.2 N proteins in E.coli BL21(DE3) at 37 DegreesCelsius for 4 hours. The indirect ELISA test showed that the titers of polyclonal antibody prepared by three N proteins were all 1:51 200. All three N proteins can increase the expression of IFN-γ and IL-1β cytokines, but the effect of Omicron N protein in activing two cytokines was more obvious than that of Wuhan-Hu-1 N protein. Conclusion The study obtained three new coronavirus N proteins and polyclonal antibodies, and confirmed that mutations in the amino acid sites of the N protein can affect its immunogenicity. This provides a basis for developing rapid diagnostic methods targeting N protein of different novel coronavirus variants.
Animals ; Mice ; SARS-CoV-2/genetics* ; Coronavirus Nucleocapsid Proteins/immunology* ; Nucleocapsid Proteins/isolation & purification* ; COVID-19/immunology* ; Antibodies, Viral/immunology* ; Mice, Inbred BALB C ; Interferon-gamma/metabolism* ; Interleukin-1beta/metabolism* ; Female ; Escherichia coli/metabolism* ; Mutation ; Humans

ObjectiveTo explore the effect of Guilu Erxianjiao on improving reproductive injury in diabetic rats and its possible mechanism. MethodFifty-three SD male rats were randomly divided into 5 groups， with 1 group as the normal group and the other 4 groups as the modeling groups. Rats in the modeling groups were fed with a high-fat diet combined with 30 mg·kg^-1 streptozotocin （STZ） intraperitoneal injection to induce diabetes， with the random blood glucose >16.7 mmol·L^-1 for 3 consecutive times as the criteria for inclusion in the model of diabetic reproductive injury. The rats with diabetic reproductive injury were then randomly divided into a model group， a Guilu Erxianjiao group （2 g·kg^-1）， a Vitamin E group （0.03 g·kg^-1）， and a Wuzi Yanzong pill group （0.6 g·kg^-1） according to the blood glucose level. The rats were given the corresponding drug dose intragastric administration， once a day for 4 weeks， and their body weight and blood glucose were measured weekly. After 4 weeks， samples were collected for index determination. Morphological changes in testis and epididymis were observed by hematoxylin-eosin （HE） staining， and apoptosis of testis cells was observed by in situ end labeling （TUNEL） staining. Sperm concentration and motility were detected by the semen analyzer. Serum levels of follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， and testosterone （T） were determined by enzyme-linked immunosorbent assay （ELISA）. The content of superoxide dismutase （SOD）， propylene glycol （MDA）， reactive oxygen species （ROS）， and glutathione peroxidase （GSH-Px） in the testicular tissue was determined by ELISA. The expressions of nuclear respiratory factor-2 （Nrf2）， heme oxygenase 1 （HO-1）， B-cell lymphoma-2 （Bcl-2）， and Bcl-2 associated X protein （Bax） in the testicular tissue were detected by Western blot. ResultAs compared with the normal group， testicular tissue atrophy， decreased spermatogenic tubules， epididymal wall hyperplasia， and lumen stenosis were observed in the model group. Sperm concentration and motility decreased （P<0.01）， and serum levels of T， FSH， and LH decreased （P<0.01） in the model group. The content of ROS and MDA in the testis increased （P<0.01）， while that of SOD and GSH-Px decreased （P<0.01） in the model group. The expression of Bax increased （P<0.01）， and the expressions of Nrf2， HO-1， and Bcl-2 decreased （P<0.01） in the model group. As compared with the model group， the pathological changes in the testis and epididymis in the Guilu Erxianjiao group were improved to some extent. Sperm concentration and motility increased （P<0.05， P<0.01）. In the Guilu Erxianjiao group， serum levels of T and LH increased （P<0.05， P<0.01）， while FSH levels showed no significant difference. The content of ROS and MDA in the testis decreased （P<0.01）， while that of SOD and GSH-Px increased （P<0.01） in the Guilu Erxianjiao group. The expression of Bax decreased （P<0.01）， and the expressions of Nrf2， HO-1， and Bcl-2 increased （P<0.05， P<0.01） in the Guilu Erxianjiao group. ConclusionGuilu Erxianjiao improves the reproductive injury and sperm quality of diabetic rats to a certain extent， and the mechanism may be related to the improvement of oxidative stress and anti-apoptosis.

Objective:To compile the evaluation scale of the quality of education in the clinical practice base of full-time Master of Specialist Nursing (MNS) based on student participation and the SERVQUAL model, evaluate the quality of clinical practice base, and provide reference for the evaluation of clinical practice base.Methods:Based on the theory of student participation and the SERVQUAL model, the evaluation scale of the educational quality of clinical practice base of MNS was constructed. By facilitating sampling methods, 176 MNS students from 11 universities in 6 provinces were selected, and a questionnaire of self-compiled general information and a survey of the evaluation scale of education quality of MNS clinical practice base were distributed.Results:The evaluation scale of education quality of clinical practice base of MNS contained 28 entries in five dimensions: tangibility, reliability, responsiveness, guarantee and empathy. The total Cronbach alpha coefficient of the scale was from 0.983 to 0.987, item level content validity index was from 0.92 to 0.96, scale level content validity index was 1 and the correlated coefficient of the structural validity Pearson was from 0.785 to 0.966. To explore factor analysis, the dimension value of the scale KMO value was greater than 0.7, and the expected part and the actual feel part KMO value were 0.956 and 0.963 respectively. The number of common factors was 2 and 3 respectively, and the cumulative interpretation variation was 79.1% and 78.4% respectively.The actual perception of the quality of education in the clinical practice base of MNS was lower than expected, and the differences in entries were statistically significant ( P<0.05). The analysis of multiple linear regression showed that the teaching time and level of clinical practice base undertaken by clinical practice base were the main factors affecting the expectation and actual feeling value ( P<0.05). The importance performance analysis analysis found that 11 entries were in the advantage zone, 3 entries were in the maintenance area, 12 entries were in the minor improvement area and 2 entries were in the priority improvement area. Conclusions:The evaluation scale of the quality of education of the clinical practice base of MNS have good confidence and validity. The actual degree of educational quality of the clinical practice base of MNS is lower than expected. The teaching time and level of clinical practice base undertaken by clinical practice base are the main factors that affect expectation and actual feeling. It is urgent to improve the theoretical knowledge and practice level of tutors and deal with occupational exposure experienced by MNS students.

Objectives To investigate the expression and distribution of two important pro-angiogenic factors, vascular endothelial growth factor (VEGF) and basic fibroblastic growth factor (bFGF), in condyloma acuminatum (CA) and their significance. Methods The expression and distribution of VEGF and bFGF in specimens from 35 cases of CA and 10 normal human skins (foreskin) were detected by streptavidin-peroxidase complex immunohistochemical technique. Results The levels of expression of VEGF and bFGF in CA were both significantly higher than those in normal skins (P﹤0.001). The levels of expression of VEGF and bFGF in CA were both throughout the epidermis but with different characteristic staining patterns, of which VEGF was intense in the basal and spinous layers and faint in the granular layer, while bFGF was faint in the basal layer and gradually became intense from spinous to granular layer and markedly increased in koilocytes. Conclusion The overexpression of VEGF and bFGF may play an important role in promoting new blood vessel formation in the dermis in CA lesions.