This study aims to obtain nanoantibody of vaccinia virus protein D8 by SUMO tag and analyze its binding activity with antigen.The sequence was designed and synthesized to construct the prokaryotic expression plasmid pKMD-SUMO-D8,which was transformed into Escherichia coli BL21(DE3)receptor cells and induced to express.Anti-SUMO-D8 with His and SUMO tags were purified,and then all tags were removed by SUMO protease to further obtain Anti-D8.Final-ly,the binding activity was detected and analyzed by indirect ELISA and IFA.The results showed that the prokaryotic expression plasmid pKMD-SUMO-D8 and recombinant strains were construc-ted.Under the condition of 30 ℃,the expression of Anti-SUMO-D8 was best when induced by IPTG with 0.1 mmol/L for 4 h.High purity Anti-SUMO-D8 and Anti-D8 nanoantibodies were ob-tained.Indirect ELISA results showed that both Anti-SUMO-D8 and Anti-D8 had binding activity with antigen.Under the same conditions,the binding activity of Anti-D8 to vaccinia virus and E8L was higher than that of Anti-SUMO-D8.Indirect IFA results showed that 48 h after infection with vaccinia virus,some nuclei of the experimental group showed green fluorescence under fluorescence microscope.Nanoantibodies of vaccinia virus protein D8 could be efficiently prepared by SUMO tag,and the prepared nanoantibodies had binding activity.This study provides new idea for the preparation of nanoantibody,and also lays a foundation for further study of the biological function of vaccinia virus protein D8 nano-antibody.

Objective:To explore the potential genetic causes of unexplained neonatal encephalopathy.Methods:This retrospective study enrolled 113 infants diagnosed with unexplained neonatal encephalopathy and underwent genetic testing in the Children's Hospital of Hunan Province from January 2019 to May 2021. Perinatal data, clinical manifestations, electroencephalograph, brain MRI findings, genetic information, and prognosis of those patients were analyzed. T-test or Chi-square test were used for data analysis. Results:Of the 113 infants enrolled, 74 (65.5%) were males. The gestational age at birth was (38.6±1.5) weeks, and the birth weight was (2 957±561) g. The most common clinical manifestation was the disturbance of consciousness (83/113, 73.5%), followed by seizures (39/113, 34.5%). There were 38.2% (34/89) of the patients with abnormal brain MRI, and 80.4% (74/92) presented abnormal electroencephalography. Among the 113 infants, 60 (53.1%) had genetic abnormalities, including 48 with single nucleotide variations, eight with copy number variations, and four with chromosome abnormalities. Single nucleotide variations in the 48 patients were classified into syndromic ( n=18, 37.5%), metabolic ( n=16, 33.3%), epileptic ( n=11, 22.9%) and mitochondrial-related genes ( n=3, 6.3%), of which 14 were not included in any database. Among the 103 cases which were successfully followed up until December 31, 2021, 75 (72.8%) had a poor prognosis, including 52 (50.5%) death cases and 23 (22.3%) cases of development retardation. Birth weight and the incidence of seizures in the poor prognosis group were both lower than those in the non-poor prognosis group [(2 876±536) vs (3 254±554) g, t=3.15; 29.3% (22/75) vs 53.6% (15/28), χ2=5.20; both P<0.05], while the incidence of disturbance of consciousness was higher [80.0% (60/75) vs 53.6% (15/28), χ2=7.19, P<0.05]. The proportion of infants with genetic abnormalities in the poor prognosis group was higher than that in the non-poor prognosis group, but the difference was not statistically significant [53.3% (40/75) vs 46.4% (13/28), χ2=0.39, P=0.533]. Conclusions:Genetic abnormality is one of the leading causes of unexplained neonatal encephalopathy. Nucleotide variation is the most common genetic type. Syndromic, metabolic, and epileptic variants are frequently detected in these patients.

Objective To explore the design of word-picture matching semantic judgment task based on Chinese high frequency nouns and further explore the way to apply the task in a functional magnetic resonance imaging (fMRI)/event-related potential (ERP) study.Methods The materials, stimuli and procedure of the word-picture matching semantic task were provided. Then, a healthy subject for the pilot study was included. In the pilot study, fMRI and ERP data of the subject were collected during the word-picture matching task. The fMRI and ERP data were analyzed to test the feasibility of the word-picture matching task in an fMRI/ERP task.Results The results of fMRI analysis showed an increased activity in the right middle frontal gyrus under the word-picture presenting condition compared with the "+" presenting condition. In addition, fMRI analysis showed an increased activity in the right middle temporal gyrus in the word-picture mismatching condition compared with the word-picture matching condition. The results of ERP analysis showed an increased activity in the left frontal/temporal area in the word-picture matching condition and an increased activity in the right frontal area in the word-picture mismatching condition.Conclusion Language associated brain regions can be identified in the fMRI/ERP research based on the word-picture matching task described in this article, which indicated that the task is effective in exploring language processing mechanism in the brain.