Objective:To construct NMB0315 eukaryotic expression recombinant vector ,detect specific humoral and cellular immune response induced by the recombint DNA vaccine intramuscularly in female BALB /c mice,evaluate the immunocompetence and immunoprotection of the vaccine , so as to provide experimental basis for the development of a novel nucleic acid vaccine against N.meningitidis serogroup B .Methods: The whole NMB0315 gene was amplified by PCR from the standard strains MC 58 genomic DNA,cloned into a plasmid pcDNA3.1(+),identified by double digestion of the recombinant plasmid with restriction enzymes and se -quencing.The recombinant vector pcDNA 3.1 (+)/NMB0315 was transfected into eukaryotic COS-7 cells and RAW264.7 cells, the NMB0315 protein was detected by immunocytochemical method and Western blot respectively .The levels of specific humoral and cellular immune response were detected after inoculating in female BALB /c mice intramuscularly with the recombinant plasmid .The immune protective effect was investigated with the DNA vaccine and the bactericidal titer of the immune serum was deter mined by serum bactericidal assay ( SBA ) in vitro.Results: The recombinant pcDNA3.1 (+)/NMB0315 was effectively transcripted and expressed in eukaryotic cells and the specific humoral and cellular immune responses were induced in the inoculated mice .In the re-combinant pcDNA3.1(+)/NMB0315 group ,the levels of serum IgG,IgG1,IgG2a,IgG2b and IgG3 and genital tract sIgA were significantly higher than in controls ( P<0.001 ) .The stimulation index in the culture supernatant of the spleen lymphocytes of the vaccine group was higher than that of the control group (P<0.05).The ratios of serum IgG2a/IgG1 in the DNA vaccine group were less than 1.The bactericidal titer of the NMB 0315+CpG group reached 1:128 following three immunizations , the protection rate of the vaccine group was 70%against the N.meningitidis strain MC58.Conclusion:The NMB0315 nucleic acid vaccine could induce higher levels of humoral immunity and cellular immunity and showed effective protection against N .meningitidis serogroup B , the immune serum had strong bactericidal activity in vitro .

Objectives:To explore the accuracy of pedicle screw placement assisted with improved 3D-print-ed drill guiding template in cervical spine.Methods:The clinical data of 60 patients undergone posterior cervical pedicle screw placement in our hospital between January 2016 and January 2023 were analyzed retro-spectively,including 30 males and 30 females with an average age of 17-84 years(58.7±13.8 years).The pa-tients were divided into guiding template group and free-hand group based on whether the guiding template was used to assist the placement of screws or not.The improved 3D-printed guiding template was used to assist cervical pedicle screw placement in the guiding template group(n=30),and the self-made angular-ruler was used in the free-hand group(n=30).There was no significant difference in age,gender ratio,and preoper-ative diagnosis between the two groups(P＞0.05).Cervical CT scan was performed at one week after surgery,and the accuracy of pedicle screw placement was evaluated according to the Kaneyama standard:Grade 0,the screw was completely in the pedicle;Grade 1,the size of screw penetrating the cortex＜50%of the screw diameter;Grade 2,the size of screw penetrating the cortex≥50%of the screw diameter but not com pletely out;Grade 3,the screw was completely on the outside of the pedicle.The accuracy of pedicle screw placement(ratio of grades 0 and 1)and complications such as vascular and nerve injury,incision infection,cerebrospinal fluid leakage,screw loosening and breakage caused by screw misplacement were recorded.Re-sults:A total of 152 pedicle screws were placed in the guiding template group,including 74 screws of grade 0,68 of grade 1,10 of grade 2 and 0 of grade 3,with an accuracy of screw placement of 93.4%.A total of 136 pedicle screws were placed in free-hand group,including 53 screws of grade 0,61 of grade 1,18 of grade 2 and 4 of grade 3,with an accuracy of screw placement of 83.8%.The accuracy of screw placement in the guiding template group was significantly higher than that in the free-hand group(P＜0.05).There were no related complications such as vascular and nerve injury,incision infection and cerebrospinal fluid leakage caused by misplacement of pedicle screws.The patients were followed up for 5-29 months(14.2±7.7months),and there were no complications such as screw loosening or breakage.Conclusions:Improved 3D-printed drill guiding template can improve the accuracy of pedicle screw placement in cervical spine.

Recently, accumulating evidence suggests that high bilirubin level is involved in the pathophysiology of schizophrenia. High bilirubin level during early childhood may increase the risk of being suffered from schizophrenia after adulthood, and schizophrenia patients with high bilirubin level have aggravated psychiatric symptoms. As compared with other psychiatric patients and general population, schizophrenia patients usually have relatively higher bilirubin level; high bilirubin level is associated with acute psychotic states, positive symptoms, and poor prognosis in patients with schizophrenia. This article reviews the relation between bilirubin and schizophrenia and its potential pathophysiological mechanism in order to provide a new direction for the study of schizophrenia pathogenesis and auxiliary diagnosis.

The rapid development of high-throughput sequencing technologies has led to a dramatic decrease in the money and time required for de novo genome sequencing or genome resequencing projects, with new genome sequences constantly released every week. Among such projects, the plethora of updated genome assemblies induces the requirement of version-dependent annotation files and other compatible public dataset for downstream analysis. To handle these tasks in an efficient manner, we developed the reference-based genome assembly and annotation tool (RGAAT), a flexible toolkit for resequencing-based consensus building and annotation update. RGAAT can detect sequence variants with comparable precision, specificity, and sensitivity to GATK and with higher precision and specificity than Freebayes and SAMtools on four DNA-seq datasets tested in this study. RGAAT can also identify sequence variants based on cross-cultivar or cross-version genomic alignments. Unlike GATK and SAMtools/BCFtools, RGAAT builds the consensus sequence by taking into account the true allele frequency. Finally, RGAAT generates a coordinate conversion file between the reference and query genomes using sequence variants and supports annotation file transfer. Compared to the rapid annotation transfer tool (RATT), RGAAT displays better performance characteristics for annotation transfer between different genome assemblies, strains, and species. In addition, RGAAT can be used for genome modification, genome comparison, and coordinate conversion. RGAAT is available at https://sourceforge.net/projects/rgaat/ and https://github.com/wushyer/RGAAT_v2 at no cost.
Genome ; Genomics ; High-Throughput Nucleotide Sequencing ; methods ; standards ; Humans ; Reference Standards ; Sequence Analysis, DNA ; methods ; standards ; Software