Objective To construct a eukaryotic expression plasmid of pcDNA 3.1-Ag85A-CD226, and to use it as DNA vaccine then further evaluate its immunogenicity through oral administration in a mouse model.Methods The CD226-PCR2.1-ToPo plasmid was used as the template to clone CD 226 gene by PCR.The CD226 gene was then inserted into pcDNA 3.1-Ag85A plasmid to construct the recombinant plas-mid of pcDNA3.1-Ag85A-CD226.After identified by restriction enzyme analysis and sequencing , the re-combinant plasmid was transfected into HEK 293 cells by using lipofection .The expression of Ag85A-CD226 gene in HEK293 cells was detected by RT-PCR, Western blot and indirect immunofluorescence assay .The purified recombinant plasmid was used to prepare the Ag 85A-CD226 DNA vaccine by liposomal encapsula-tion.The vaccine was administered intragastrically to mice .The activities of NK cells , the cytokine levels in the supernatants of spleen cell cultures and the mRNA level of cytokines in the intestines were evaluated to analyze the immunogenicity of Ag85A-CD226 DNA vaccine.Results The Ag85A-CD226 DNA vaccine was prepared successfully .The expression of Ag85A-CD226 fusion protein was detected in HEK293 cells.The activities of NK cells from mice vaccinated with Ag 85A-CD226 DNA vaccine were higher than those from other control groups (P<0.01).The level of TNF-α, IFN-γand IL-2 in the supernatants of spleen cell cul-tures and in the intestines were significantly up-regulated in comparison with other control groups ( P <0.01).The level of IL-4 in the supernatants of spleen cell cultures was down-regulated in the experimental group (P<0.01), but the level of IL-4 in intestines showed no significant difference among the five groups (P>0.05).Conclusion The Ag85A-CD226 DNA vaccine could significantly enhance Th1 type immune responses systemically and in the intestine as in comparison with those vaccinated with single dose of Ag 85A DNA vaccine or CD226 DNA vaccine.

Objective To explore the incidence and risk factors of acute kidney injury (AKI) following hepatic arterial chemoembolization inpatients with hepatocellular carcinoma(HCC),Child class A. Methods Retrospective analysis was performed on patients with HCC,Child Class A who were admitted from January 2008 to October 2016.The inclusion criteria:(1) patients were diagnosed with pathologically confirmed hepatocellular carcinoma based on the standards of primary liver cancer diagnosis and treatment (2011 edition);(2) patients have complete laboratory parameters and imaging data within the 7 days before the TACE treatment;(3) the Child-pugh score was 5 to 6;(4) patients have no acute or chronic renalfailure before the treatment of TACE.The morbidity of AKI after TACE was investigated.Twenty-six factors such as age,sexy,contrast dose,hemoglobin were included as independent variable, so as to investigate the risk factors for postoperative AKI. Results A total of 818 patients were included in the study, including 38 experiencing postoperative AKI(4.64%).Multivariate logistic regression analysis identified elevated preoperative uric acid as the independent risk factor for postoperative AKI(OR=1.005,95% CI 1.000 to 1.009,P=0.037), and elevated preoperative hemoglobin as the protective factor for postoperative AKI(OR=0.974,95% CI 0.952 to 0.997,P=0.028).Conclusions AKI is not uncommon following hepatic arterial chemoembolization inpatients with HCC,Child Class A,and is correlated with elevated uric acid and anemia.Preoperative risk assessment and anemia correction might be effective for reducing the incidence of AKI following hepatic arterial chemoembolization.

Objective To investigate the effect of improving immunity in the treatment of chronic brucellosis,and to analyze and evaluate its clinical curative effect.Methods A patient with chronic brucellosis was treated with Mongolian medicine combined with chemical drugs to enhance immunity.The clinical symptoms,serological antibodies,Brucella DNA and immune function were compared before and after treatment.The specific antibody against Brucella in serum was detected by tube agglutination test (SAT) and tiger red plate agglutination test (RBPT).Brucella DNA in serum and blood cells was detected by PCR,and the peripheral blood lymphocyte subsets were detected by flow cytometry.Immuno-luminescence technique was used to detect serum immunoglobulin and complement components.Results After treatment,the clinical symptoms such as cold back,fatigue,and joint pain disappeared completely,and the results of serum specific antibodies against Brucella were SAT 1 ∶ 50 (++)and RBPT (+) with no changes before and after treatment,and the results of cells and serum were both negative after treatment though the results of DNA detection of Brucella were cell positive and seronegative before treatment.The results of immunological function test showed that γδT cells decreased to 9.50％ after treatment compared to 14.00％ before treatment,and the percentage of monocytes and Treg cells were 5.59％ and 7.33％ after treatment,which were higher than 3.35％ and 4.72％ of before treatment,and the level of complement C3 was 0.79 g/L before treatment and 0.91 g/L after treatment that was returned to normal reference range (0.88 ～ 2.01 g/L).Conclusion The patients with chronic brucellosis can improve their clinical treatment by improving immunity.

Objective:To investigate the clinical manifestations of population following COVID-19 by using questionnaires.Methods:COVID-19 among anesthesia workers and the surrounding population was investigated between 11 November 2022 and 31 December 2022 in China.The Tencent electronic questionnaire(ID.11492813) was sent to different WeChat groups of the Association of Anesthesiologists or Society of Anesthesiologists via the WeChat platform of the medical personnel in China. The survey was conducted between January 7 and January 15, 2023. Results:A total of 17 000 questionnaires were issued for this survey, 11 060 valid questionnaires from 31 provinces and autonomous regions were collected all over the country, with a recovery rate of 65.059%.There were 10068 (91.037%) participants diagnosed as having COVID-19, and among of them, 47.606% were male and 52.394% were female. The main post-COVID-19 clinical manifestations included fever (85.777%), cough (83.731%), fatigue (75.338%), parasomnia (64.352%), limb soreness (58.890%), dizziness, headache, tinnitus (38.617%), loss or abnormality of taste (37.763%), and loss or abnormality of smell (30.960%); peripheral neuralgia was usually found within 3 days after positive nucleic acid test or positive antigen test; there were 2 963 cases accompanied with sweating, and among of them, 47.25% were male and 52.75% were female, and 37.80% of these participants continued to sweat after the nucleic acid test or antigen test became negative. There were 1 151 cases with premature heart beats among the study population, and the symptoms aggravated following COVID-19 were found in 34.32% of these patients.Conclusions:In addition to the respiratory system, the central and peripheral nerves of patients are also affected following COVID-19, and the peripheral and central nerve disorders last until several days after negative nucleic acid test or antigen test, suggesting that anesthesiologists should pay more attention to monitoring of various nerve function and impact of surgery and anesthetic drugs on the stress response of the body in such patients.