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

OBJECTIVE To retrospectively analyze the clinical efficacy of Qingchi San in the treatment of mild-to-moderate ul-cerative colitis.METHODS A retrospective analysis was conducted on 221 ulcerative colitis patients treated at the Gastroenterology Department of First Teaching Hospital of Tianjin University of Traditional Chinese Medicine from December 2014 to February 2024.Participants were divided into an observation group(n=121)and a control group(n=100).The control group received oral me-salazine,while the observation group received Qingchi San enema in addition to the control group's treatment.Both groups received a 4-week course of treatment.The two groups were compared in terms of clinical efficacy,clinical remission rate,TCM syndrome score,time to intestinal bleeding remission,and changes in inflammatory markers(C-reactive protein,erythrocyte sedimentation rate,and white blood cells)and coagulation markers(platelets and D-dimer).RESULTS After treatment,the total effective rate and clinical remission rate in the observation group were significantly higher than those in the control group(P<0.05,P<0.01).The TCM syn-drome scores for each item(diarrhea,abdominal pain,bloody stools with mucus,and tenesmus)in the observation group were signifi-cantly improved compared with those in the control group(P<0.01).The time to intestinal bleeding remission in the observation group was significantly shorter than that in the control group(P<0.01).C-reactive protein,erythrocyte sedimentation rate,and white blood cell count in the observation group were significantly decreased compared with those in the control group(P<0.05,P<0.01).There was no statistically significant difference in coagulation markers(platelets and D-dimer)between the two groups(P>0.05).No signif-icant treatment-related adverse reactions,such as liver and kidney damage,were observed during treatment.CONCLUSION Qing-chi San enema can improve the clinical efficacy of patients with mild-to-moderate ulcerative colitis,improve clinical symptoms,short-en bleeding time,and reduce inflammatory markers.

OBJECTIVE To retrospectively analyze the clinical efficacy of Qingchi San in the treatment of mild-to-moderate ul-cerative colitis.METHODS A retrospective analysis was conducted on 221 ulcerative colitis patients treated at the Gastroenterology Department of First Teaching Hospital of Tianjin University of Traditional Chinese Medicine from December 2014 to February 2024.Participants were divided into an observation group(n=121)and a control group(n=100).The control group received oral me-salazine,while the observation group received Qingchi San enema in addition to the control group's treatment.Both groups received a 4-week course of treatment.The two groups were compared in terms of clinical efficacy,clinical remission rate,TCM syndrome score,time to intestinal bleeding remission,and changes in inflammatory markers(C-reactive protein,erythrocyte sedimentation rate,and white blood cells)and coagulation markers(platelets and D-dimer).RESULTS After treatment,the total effective rate and clinical remission rate in the observation group were significantly higher than those in the control group(P<0.05,P<0.01).The TCM syn-drome scores for each item(diarrhea,abdominal pain,bloody stools with mucus,and tenesmus)in the observation group were signifi-cantly improved compared with those in the control group(P<0.01).The time to intestinal bleeding remission in the observation group was significantly shorter than that in the control group(P<0.01).C-reactive protein,erythrocyte sedimentation rate,and white blood cell count in the observation group were significantly decreased compared with those in the control group(P<0.05,P<0.01).There was no statistically significant difference in coagulation markers(platelets and D-dimer)between the two groups(P>0.05).No signif-icant treatment-related adverse reactions,such as liver and kidney damage,were observed during treatment.CONCLUSION Qing-chi San enema can improve the clinical efficacy of patients with mild-to-moderate ulcerative colitis,improve clinical symptoms,short-en bleeding time,and reduce inflammatory markers.

Selenium is an essential trace element for human body.The primary functional form of selenium in the organism is selenoproteins(Sep),which plays a crucial role in maintaining normal physiological functions,influencing immune responses,and is also implicated in tumorigenesis and related metabolic disorders.Recent studies have shown that Sep is involved in the pathophysiological process of inflammatory bowel disease(IBD).This article reviewed the mechanisms of Sep in IBD,aiming to explore the pathogenic mechanisms of IBD and provide insights for the diagnosis and treatment of the disease.

Selenium is an essential trace element for human body.The primary functional form of selenium in the organism is selenoproteins(Sep),which plays a crucial role in maintaining normal physiological functions,influencing immune responses,and is also implicated in tumorigenesis and related metabolic disorders.Recent studies have shown that Sep is involved in the pathophysiological process of inflammatory bowel disease(IBD).This article reviewed the mechanisms of Sep in IBD,aiming to explore the pathogenic mechanisms of IBD and provide insights for the diagnosis and treatment of the disease.

Objective To explore the effect of cannabinoid receptor 2(CB2)on orthodontic tooth movement(OTM)rate and periodontal tissue reconstruction of pressure area in mice.Methods Thirty CB2-/-male mice and thirty littermate control WT male mice were individually accepted the orthodontic appliance at their age of 6 weeks.The mice were respectively scarified at 3 days,7 days,14 days and 21 days after the operation.Then the tooth movement distance was examined through the stereomicroscope.Hematoxylin-eosin staining was performed to explore the biological responses of periodontium at the distal mesial root pressure area.Anti-tartrate acid phospha-tase staining was performed to calculate the number and distribution of osteoclasts at the distal mesial root pressure area,and MMP-9 was evaluated by immunohistochemistry to examine the number of MMP-9(+)monocytes and multinucleated cells in the same district as the TRAP staining.Results Compared with those WT mice at 3,7,14 and 21 days,OTM distance showed a gradual increased tendency according with experimental time over 21 days.The widths of periodontal ligament on the pressure side were markedly greater in CB2-/-mice than WT mice at 7,14 and 21 days(P＜0.000 1).The numbers of TRAP positive osteoclasts were significantly greater in CB2-/-mice than those in WT mice at 14 days of OTM(P＜0.001).MMP-9 immunohistochemical staining showed that the number of MMP-9(+)monocytes and multinucleated cells was more in CB2-/-mice than that in WT mice at 14 days of OTM(P＜0.05).Conclusion The absence of CB2 accelerates orthodontic tooth movement under or-thodontic force.The absence of CB2 reinforces bone resorption in orthodontic tooth movement compressive area dur-ing orthodontic tooth movement.

Objective:To discuss the effect of downregulating the proline-rich protein 11(PRR11)expression on drug resistance of the esophageal cancer drug resistant cells,and to clarify the related mechanism.Methods:The drug resistant cells EC9706/cisplatin(DDP)were established by incrementally stimulating the human esophageal cancer EC9706 cells with the increasing concentrations of DDP.The drug sensitivity of the EC9706/DDP cells was detected by MTT assay;the expression levels of PRR11 mRNA and protein in the EC9706/DDP cells and their parent EC9706 cells were detected by real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods.The EC9706/DDP cells were divided into control group,sh-NC group(infected with sh-NC),sh-PRR11 group(infected with sh-PRR11),sh-NC+DDP group(infected with sh-NC and treated with 4 mg·L-1 DDP),and sh-PRR11+DDP group(infected with sh-PRR11 and treated with 4 mg·L-1 DDP).The expression levels of PRR11 mRNA in the cells in various groups were detected by RT-qPCR method;the expression levels of PRR11,phosphoinositide 3-kinase(PI3K)p110α,protein kinase B(AKT),phosphorylated AKT(p-AKT),P-glycoprotein(P-gp),and multidrug resistance-associated protein 1(MRP1)proteins in the cells in various groups were detected by Western blotting method;the apoptotic rates of the cells in various groups were detected by flow cytometry.Results:The DDP-resistant cell line EC9706/DDP was successfully obtained,and the drug resistance index was 7.23±0.86.Compared with the EC9706 cells,the expression levels of PRR11 mRNA and protein in the EC9706/DDP cells were increased(P<0.05).Compared with control and sh-NC groups,the expression levels of PRR11 mRNA and protein in the cells in sh-PRR11 group were decreased(P<0.05),and the 50%inhibitory concentration(IC50)of DDP was decreased(P<0.05).Compared with sh-NC group,the expression levels of PI3K p110α,p-AKT,P-gp,and MRP1 proteins in the cells in sh-NC+DDP and sh-PRR11 groups were decreased(P<0.05),and the apoptotic rate of the cells was increased(P<0.05).Compared with sh-NC+DDP group and sh-PRR11 group,the expression levels of PI3K p110α,p-AKT,P-gp,and MRP1 proteins in the cells in sh-PRR11+ DDP group were increased(P<0.05),and the apoptotic rate of the cells was increased(P<0.05).Conclusion:Downregulating the expression of PRR11 gene in the drug resistant EC9706/DDP cells can inhibit the expressions of drug resistance-related proteins,reverse the resistance to DDP,and induce the apoptosis;its mechanism may be related to the inhibition of activation of the PI3K/AKT signaling pathway.

Objective:To investigate the clinical and genetic characteristics of families with myotonic dystrophy type I.Methods:Two families with myotonic dystrophy type I admitted to Department of Neurology, First Affiliated Hospital of He'nan University of Science and Technology in September and October 2021 were chosen; their clinical data were retrospectively analyzed. The muscle pathological changes were confirmed by electromyography and muscle MRI. Repeat-primed PCR assay was used to detect the number of CTG repeats in the 3' end non-coding region of DMPK gene. Results:Clinical heterogeneity existed among the two families of patients; muscle weakness and muscular atrophy of the skeletal muscle were the main clinical manifestations; limb weakness, axe face, percussion myotonia and myoglobular sign were noted; systemic multi-system symptoms included palpitation and chest tightness, cataracts, gastrointestinal symptoms, fatigue/lethargy, and cognitive impairment. Electromyography showed myotonic potential and myogenic damage. Muscle fatty infiltration and atrophy were noted in muscle MRI, and lesions were predominantly in the gastrocnemius. All patients had abnormal amplification of DMPK gene CTG (number of CTG repeats> 50 or 100). Conclusion:In addition to skeletal muscle involvement, systemic multi-system involvement such as cardiac, eye, respiratory, endocrine, and nervous system should also be noted by clinicians in patients with myotonic dystrophy type I.

Objective:To investigate the factors affecting phenotypes in the patients of methylmalonic acidemia combined with homocysteinemia cblC type with MMACHC c. 609G>A homologous variant. Methods:A retrospective study on the clinical manifestations, complications, treatment, and outcome in 164patients of cblC type with MMACHC c. 609G>A homologous variant was conducted.The patients were diagnosed by biochemical and genetic analysisfrom January 1998 to December 2020. Results:Among the 164 patients, 2 cases were prenatally diagnosed and began treatment after birth. They are 3 and 12 years old with normal physical and mental development. Twenty-one cases were diagnosed by newborn screening. Among them, 15 cases had with normal development. They were treated fromthe age of two weeks at the asymptomatic period. Six cases began treatment aged 1 to 3 months after onset. Their development was delayed. One hundred and forty-one cases were clinically diagnosed. Their onset age ranges from a few minutes after birth to 6 years old. 110 cases had early-onset (78.0%). 31 cases had late-onset (22.0%). Five of them died. 24 patients lost to follow-up. Of the 141 clinically diagnosed patients, 130 (92.2%) with psychomotor retardation, 69 (48.9%) with epilepsy, 39 (27.7%) with anemia, 30 (21.3%) had visual impairment, 27 (19.1%) had hydrocephalus, 26 (18.4%) had feeding difficulties, 7 (5.0%) with liver damage, and 5 (3.5%) with metabolic syndrome. The frequency of hydrocephalus and seizures was significantly higher in the early-onset group. The urinary methylmalonic acid increased significantly in the patients with epilepsy. During the long-term follow-up, the level of plasma total homocysteine in the seizure-uncontrolled group was significantly higher than that in the seizure-controlled group, the difference had a statistical significance ( P<0.05). Conclusion:Most of the patients with MMACHC c. 609G>A homozygous variant had early-onset disease, with a high mortality and disability rate. If not treated in time, it will lead to neurological damage, resulting in epilepsy, mental retardation, hydrocephalus, and multiple organ damage. Pre-symptomatic diagnosis and treatment are crucial to prevent irreversible neurological damage. Neonatal screening and prenatal diagnosis are important to improve the outcome of the patients.

Objective:To study the effect of dexmedetomidine (Dex) combined with sevoflurane anesthesia on postoperative cognitive function and hemodynamics in elderly patients with humeral fractures during the operation.Methods:A total of 120 elderly patients with humeral fractures diagnosed and treated in Chaoyang Central Hospital from January 2018 to June 2020 were selected and divided into the control group (59 cases) and the study group (61 cases) according to the non-randomized clinical concurrent controlled study and patients′ voluntary principle. Sevoflurane inhalation was given to patients to maintain general anesthesia in two groups, while Dex anesthesia was given to the study group. Cognitive function, hemodynamics, stress response and pain were compared between the two groups before and after the operation, and anesthesia related complications were observed and recorded.Results:The mean arterial pressure (MAP) in the study group at 10 min after endotracheal intubation (T 1), operation time (T 2), 30 min after the beginning of the operation (T 3) were lower than those in the control group, the heart rate (HR) in the study group at T 2, T 3 and the immediate time after the operation (T 4) were lower than those in the control group, the differences were statistically significant ( P<0.05). The levels of norepinephrine (NE) and cortisol (Cor) in the study group were lower than those in the control group after the surgery: (0.92 ± 0.19) mmol/L vs. (1.10 ± 0.28) mmol/L, (213.69 ± 20.83) μg/L vs. (258.43 ± 28.27) μg/L, the differences were statistically significant ( P<0.05). The scores of Montreal Cognitive Assessmentin the study group on 3, 7 d after the operation were higher than those in the control group: (23.42 ± 1.37) points vs. (21.39 ± 1.53) points, (25.83 ± 0.95) points vs. (25.14 ± 0.99) points, the differences were statistically significant ( P<0.05). The incidence of anesthesia related complications in the study group was lower than that in the control group: 8.2%(5/61) vs. 22.0%(13/59), χ2 = 4.50, P<0.05. Conclusions:The application of Dex combined with sevoflurane in the anesthesia of elderly patients with humeral fractures is beneficial to maintain hemodynamic stability, reduce stress response and pain degree, and promote the recovery of cognitive function.