This study aims to investigate the isolate and identify of infectious bronchitis virus(IBV)in chickens,and study its genetic variation and pathogenicity.In 2023,two strains named CK/CH/HN/SQ202301 and CK/CH/HN/SQ202302 were obtained from suspected infectious bronchitis(IB)infected materials collected in a region of Henan Province,China.Further analysis showed that the two isolates belong to the G Ⅰ-13 and GⅥ genotypes,respectively.The cleavage sites of S protein were all RRSRR.The prediction of glycosylation sites showed that the two isolates had 18 and 12 N-glycosylation sites respectively,but no O-glycosylation site.Recombinant analysis shows that C2023-1 was a recombinant strain.Pathogenicity was assessed by infecting 1-day-old SPF chicks with the two isolates,and the results showed that C2023-1 strain infection could cause clini-cal symptoms such as depression and head shaking,as well as death in chicks,with a mortality rate of 37.5％.There were no clinical symptoms or deaths after infection with C2023-2 strain.Viral load test results showed that both isolates continued to detoxify until the 10th day,and had strong rep-lication capacity in the kidney,trachea and bursa of Fabricius.The results indicate significant differences in the genetic characteristics and pathogenicity of the two isolates due to their different genotypes.This study not only provides new epidemiological data on IB,which contributes to a bet-ter understanding of IBV's epidemiological features and control challenges,but also adds valuable bioinformatics resources for IBV by analyzing its variation mechanisms and biological information.

To verify whether the two B-cell epitopes Pep1 and Pep4 in the FAdV-4 WZ fiber can be used as candidate epitopes for multivalent epitope vaccines,epitopes Pep1 and Pep4 were tandemly linked with the chicken infectious bronchitis virus strain M41 S1 protein gene in different patterns,and a recombinant fusion plasmid was constructed and expressed in E.coli BL21(DE3).It was confirmed by Western blot and ELISA tests that all four expressed fusion proteins reacted specific-ally with anti-M41 whole virus serum and WZ strain anti-Fiber 2-knob protein serum.After purifi-cation and immunization of BALB/c mice,specific antibodies against the peptide epitopes were de-tected in mouse sera.The results showed that the Pep4 epitope induced a stronger immune re-sponse than the Pep1 epitope.When Pep1 was connected with the amino and carboxyl termini of the fusion protein,respectively,both resulted in the production of the same level of anti-Pep1 anti-bodies in the immunized animals,whereas when Pep4 was connected with the carboxyl terminus of the fusion protein,the immunized animals produced a higher level of anti-Pep4-specific antibodies.This research indicates that the B cell epitopes Pep1 and Pep4 of the reactive WZ strain Fiber 2,when conjugated with proteins to form fusion proteins,can enhance the immunogenicity of Pep1 and Pep4 without affecting the antigenicity of the carrier protein.This study provides technical support and serves as a reference for the design and development of a multivalent epitope vaccine for FAdV-4.

To verify whether the two B-cell epitopes Pep1 and Pep4 in the FAdV-4 WZ fiber can be used as candidate epitopes for multivalent epitope vaccines,epitopes Pep1 and Pep4 were tandemly linked with the chicken infectious bronchitis virus strain M41 S1 protein gene in different patterns,and a recombinant fusion plasmid was constructed and expressed in E.coli BL21(DE3).It was confirmed by Western blot and ELISA tests that all four expressed fusion proteins reacted specific-ally with anti-M41 whole virus serum and WZ strain anti-Fiber 2-knob protein serum.After purifi-cation and immunization of BALB/c mice,specific antibodies against the peptide epitopes were de-tected in mouse sera.The results showed that the Pep4 epitope induced a stronger immune re-sponse than the Pep1 epitope.When Pep1 was connected with the amino and carboxyl termini of the fusion protein,respectively,both resulted in the production of the same level of anti-Pep1 anti-bodies in the immunized animals,whereas when Pep4 was connected with the carboxyl terminus of the fusion protein,the immunized animals produced a higher level of anti-Pep4-specific antibodies.This research indicates that the B cell epitopes Pep1 and Pep4 of the reactive WZ strain Fiber 2,when conjugated with proteins to form fusion proteins,can enhance the immunogenicity of Pep1 and Pep4 without affecting the antigenicity of the carrier protein.This study provides technical support and serves as a reference for the design and development of a multivalent epitope vaccine for FAdV-4.

This study aims to investigate the isolate and identify of infectious bronchitis virus(IBV)in chickens,and study its genetic variation and pathogenicity.In 2023,two strains named CK/CH/HN/SQ202301 and CK/CH/HN/SQ202302 were obtained from suspected infectious bronchitis(IB)infected materials collected in a region of Henan Province,China.Further analysis showed that the two isolates belong to the G Ⅰ-13 and GⅥ genotypes,respectively.The cleavage sites of S protein were all RRSRR.The prediction of glycosylation sites showed that the two isolates had 18 and 12 N-glycosylation sites respectively,but no O-glycosylation site.Recombinant analysis shows that C2023-1 was a recombinant strain.Pathogenicity was assessed by infecting 1-day-old SPF chicks with the two isolates,and the results showed that C2023-1 strain infection could cause clini-cal symptoms such as depression and head shaking,as well as death in chicks,with a mortality rate of 37.5％.There were no clinical symptoms or deaths after infection with C2023-2 strain.Viral load test results showed that both isolates continued to detoxify until the 10th day,and had strong rep-lication capacity in the kidney,trachea and bursa of Fabricius.The results indicate significant differences in the genetic characteristics and pathogenicity of the two isolates due to their different genotypes.This study not only provides new epidemiological data on IB,which contributes to a bet-ter understanding of IBV's epidemiological features and control challenges,but also adds valuable bioinformatics resources for IBV by analyzing its variation mechanisms and biological information.

"Weibing" is a fundamental concept in traditional Chinese medicine (TCM), representing a transitional state characterized by diminished self-regulatory abilities without overt physiological or social dysfunction. This perspective delves into the biological foundations and quantifiable markers of Weibing, aiming to establish a research framework for early disease intervention. Here, we propose the "Health Quadrant Classification" system, which divides the state of human body into health, sub-health, disease-susceptible state, and disease. We suggest the disease-susceptible stage emerges as a pivotal point for TCM interventions. To understand the intrinsic dynamics of this state, we propose laboratory and clinical studies utilizing time-series experiments and stress-induced disease susceptibility models. At the molecular level, bio-omics technologies and bioinformatics approaches are highlighted for uncovering intricate changes during disease progression. Furthermore, we discuss the application of mathematical models and artificial intelligence in developing early warning systems to anticipate and avert the transition from health to disease. This approach resonates with TCM's preventive philosophy, emphasizing proactive health maintenance and disease prevention. Ultimately, our perspective underscores the significance of integrating modern scientific methodologies with TCM principles to propel Weibing research and early intervention strategies forward.

Objective To investigate the effects of artesunate(Art)on the proliferation,apoptosis,and autophagy of nephroblastoma cell line(SK-NEP-1).Methods SK-NEP-1 cells were intervened with different concentrations of Art(0,10,20,40 and 80 μmol/L),and MTT method was applied to calculate the cell proliferation inhibition rate to screen the optimal intervention concentration;SK-NEP-1 cells were separated into control group,Art group,3-MA group(Art+autophagy inhibitor,3-methyladenine),and Rapa group(Art+autophagy activator rapamycin).EdU and flow cytometry were applied to detect cell proliferation and apoptosis,respectively;MDC staining was applied to detect autophagy in cells;the level of reactive oxygen species(ROS)in cells was detected by DCFH-DA fluorescent probe;the expression of proliferating cell nuclear antigen(PCNA),anti apoptotic factor B cell lym-phomatoma-2(Bcl-2),Bcl-2 associated X protein(Bax),microtubule junction protein 1 light chain 3 Ⅱ/3 Ⅰ(LC3 Ⅱ/LC3 Ⅰ),selective autophagy junction protein 1(p62),and benzyl chloride 1(Beclin-1)proteins in cells were detected by Western blot.Results Compared with 0 μmol/L Art,the proliferation inhibition rate of SK-NEP-1 cells was gradually increased after 10,20,40 and 80 μmol/L Art treatment(P＜0.05),and the IC50 value was 46.881 μmol/L,so 40 μmol/L Art was selected for follow-up experiments.Compared with the control group,the apoptosis rate,relative autophagy fluorescence intensity,ROS level,Bax,LC3 Ⅱ/LC3 Ⅰ,Beclin-1,PINK1,and Parkin protein expression levels of SK-NEP-1 cells in the Art group were obviously increased,the EdU positive cell rate,PCNA,Bcl-2,and P62 protein expression levels were obviously reduced(P＜0.05);The auto-phagy inhibitor 3-MA inhibited the promoting effect of Art on apoptosis and autophagy of nephroblastoma cells and inhibit proliferation(P＜0.05).Conclusions Art inhibits the proliferation of nephroblastoma cell line SK-NEP-1,and promotes autophagy and apoptosis.

Objective To explore the feasibility of electrical impedance tomography(EIT)for real-time and accurate monitoring of respiration during HP anti-G movements and the key parameters of pulmonary ventilation.Methods Twelve healthy male students in our university were enrolled in September 2023 and subjected in this study.HP anti-G movements were performed 3 times each for 30 s in the anti-G physiological training apparatus,during which EIT and ophthalmic horizontal arterial pressure were measured to analyze the relationship of the global and local parameters of pulmonary ventilation,including inspiratory volume(IV),expiratory uniform(EU),expiratory speed(ES),center of ventilation(COV)and right-to-left lung ventilation ratio(RtoL)with anti-G ability of anti-G straining maneuver(AGSM).Results The average eye horizontal systolic blood pressure(SBP)at the eye level was 148.82±22.75 mmHg during HP anti-G movements,which was significantly higher than that during quiet breathing(PJ)(95.17±8.51 mmHg,P＜0.001).From the global pulmonary ventilation,the participants had significantly increased IV during HP anti-G movements(P＜0.001).According the d value(mean increase of eye horizontal SBP=SBPHP-SBPPJ),the subjects were divided into 3 groups,with the d value of＞60,30～60 and＜30 mmHg,respectively.The inspiratory volume ratio(IVHP/IVPJ)was the highest in the＞60 mmHg group and the smallest in the＜30 mmHg group(P＜0.01).The subjects had significantly decreased EU and more evenly expiration(P＜0.05),but no change was seen in the expiratory uniformity ratio(EUHP/EUPJ)among the 3 groups.ES was obviously faster during HP anti-G movements(P＜0.001),and the expiratory speed ratio(ESHP/ESPJ)had no significant difference among the 3 groups.The inspiratory time and expiratory time were 0.77±0.32 and 1.59±0.21 s,respectively,and both of them were notably shorter during HP anti-G movements(P＜0.001,P＜0.01).From the local pulmonary ventilation,COV during HP anti-G movements was significantly smaller than that during PJ(P＜0.001),and the ventilation center deviated to the ventral side,and RtoL was decreased and the ventilation distribution deviated to the left lung(P＜0.05).Conclusion EIT can perform real-time imaging of pulmonary global and local ventilation during HP anti-G movements,and it has a great application prospect in AGSM training and monitoring.

Objective:To investigate the clinical effect of anterior cervical release and posterior fixation in the treatment of irreducible atlantoaxial dislocation with retropharyngeal internal carotid artery.Methods:Thirteen patients with irreducible atlantoaxial dislocation of retropharyngeal internal carotid artery from January 2015 to July 2019 were treated with anterior cervical release and posterior fixation. There were 8 males and 5 females, aged from 34 to 65 years with an average of 46.1±12.6 years. Positive, lateral and dynamic X-ray films, MR and CTA were performed before operation. There were 4 cases with bilateral retropharyngeal internal carotid artery and 9 cases with unilateral retropharyngeal internal pharyngeal artery. The time of operation, the amount of bleeding and intraoperative and postoperative complications were recorded. The main observations were Japanese Orthopaedic Association (JOA) score, atlantodental interval (ADI), Chamberlain line (CL), and changes in the morphology of the retropharyngeal internal carotid artery and implant fusion.Results:All the operations completed successfully. The operation time was 210-260 min, the average was 245±21 min; the blood loss was 350-600 ml, the average blood loss was 490±107 ml. There was no injury of internal carotid artery, vertebral artery, spinal cord or nerve root during the operation. All patients were followed up for 9 to 24 months, with an average of 15.1±6.2 months. Preoperative JOA score was 6.9±2.3 points, 1 month after operation was 13.5±2.5 points, and the last follow-up was 14.3±2.1 points. The difference was statistically significant ( F=30.91, P<0.001). The difference between 1 month after operation and before operation was statistically significant ( P<0.001), and the improvement rate of JOA score was 75.6%±15.2%. There was no significant difference between the last follow-up and 1 month after operation ( P>0.05). The preoperative ADI was 8.9±2.2 mm, 1 month after operation was 1.1±0.8 mm, and the last follow-up was 1.2±0.9 mm. The difference was statistically significant ( F=114.69, P<0.001). The difference between 1 month after operation and before operation was statistically significant ( P<0.001), and ADI had returned to normal level. There was no significant difference between the last follow-up and 1 month after operation ( P>0.05). The preoperative CL was 11.7±4.8 mm, 1 month after operation was 1.6±2.1 mm, and the last follow-up was 1.8±2.3 mm. The difference was statistically significant ( F=34.19, P<0.001). The difference between 1 month after operation and before operation was statistically significant ( P<0.001), and the position of odontoid process returned to normal level. There was no significant difference between the last follow-up and 1 month after operation ( P>0.05). Bone graft fusion was received at 6 to 12 months after operation, with an average of 10.2 months. Conclusion:CTA examination should be performed before anterior release of atlantoaxial dislocation to understand the position and shape of internal carotid artery. Anterior cervical release combined with posterior bone graft fusion is an effective method for the treatment of irreducible atlantoaxial dislocation with retropharyngeal internal carotid artery without increasing the risk of internal carotid artery injury.

Objective:To analyze the expression of cell differentiation cyclin 45 (CDC45) in hepatocellular carcinoma (HCC) tissues, to explore the relationship between CDC45 and the prognosis of HCC, and to explore the possibility of CDC45 as a potential biomarker and therapeutic target for HCC.Methods:Bioinformatics methods were used to analyze the mRNA levels of CDC45 in HCC tissues and normal tissues, and to analyze the relationship between its expression and the survival rate of HCC patients. The clinicopathological data of 90 patients with HCC undergoing surgical treatment were retrospectively analyzed. Immunohistochemical method was used to detect the expression level of CDC45 protein in HCC tissue and normal tissues, and to analyze its relationship with clinicopathological characteristics of patients with HCC.Results:CDC45 is highly expressed in colorectal cancer, breast cancer, prostate cancer, lung cancer, liver cancer, etc. The expression of CDC45 in HCC tissues was higher than that in normal tissues adjacent to cancer ( P<0.05). The expression of CDC45 is significantly related to the clinical prognosis of HCC patients ( P<0.001), and the higher the expression of CDC45, the lower the disease-free survival rate and overall survival rate of HCC patients. CDC45 level is significantly correlated with the number of tumor nodules ( P=0.003) and tumor size ( P=0.003) in HCC patients, but the correlation between CDC45 level and age, gender and alpha-fetoprotein (AFP) levels is not statistically significant (all P>0.05). Conclusions:CDC45 plays an important role in the progression of HCC and is expected to become a therapeutic target and potential biomarker for HCC.

A chemical investigation on