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Chinese Journal of Biotechnology ; (12): 2066-2075, 2020.
Article in Chinese | WPRIM | ID: wpr-878466


To achieve uniform soluble expression of multiple proteins in the same Escherichia coli strain, and simplify the process steps of antigen production in genetic engineering subunit multivalent vaccine, we co-expressed three avian virus proteins including the fowl adenovirus serotype 4 (FAdV-4) Fiber-2 protein, infectious bursal disease virus (IBDV) VP2 protein and egg-drop syndrome virus (EDSV) Fiber protein in E. coli BL21(DE3) cells after optimization of gene codon, promoter, and tandem expression order. The purified proteins were analyzed by Western blotting and agar gel precipitation (AGP). The content of the three proteins were well-proportioned after co-expression and the purity of the purified proteins were more than 80%. Western blotting analysis and AGP experiment results show that all the three co-expression proteins had immunoreactivity and antigenicity. It is the first time to achieve the three different avian virus antigens co-expression and co-purification, which simplified the process of antigen production and laid a foundation for the development of genetic engineering subunit multivalent vaccine.

Animals , Antigens, Viral/genetics , Biological Assay , Chickens/immunology , Escherichia coli/genetics , Infectious bursal disease virus/immunology , Poultry Diseases , Vaccines, Synthetic/isolation & purification , Viral Structural Proteins/immunology , Viral Vaccines/immunology
Article in Chinese | WPRIM | ID: wpr-669286


Objective To discuss the effects of lidocaine infusion on perioperative immune function by evaluating the levels of stress hormone and natural killer (NK) cell cytotoxicity.Methods Thirty-five patients of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,aged 35-65 yr,undergoing elective radical hysterectomy,were randomized into lidocaine group (group L)and control group (group C).Fifteen minutes before anesthesia induction,a bolus of 1.5 mg/kg of lidocaine was administered each patient in group L and followed by a continuous infusion at 1.5 mg·kg-1 ·h-1 lasting to the end of surgery.Meanwhile,the patients in group C received the same volume of saline.Venous blood samples were collected individually 24 h before the operation,the end of the operation and 48 h after the operation.Levels of prostaglandin,epinephrine and norepinephrine were assayed by ELISA kits.NK Cells were obtained by CD56 antibody magnetic isolation.The cytotoxicity of NK cell was detected by LDH releasing assay,and phosphor-protein kinase A (p-PKA)and protein kinase A (PKA) were detected by Western blotting.Results There were no significantly different in the plasm levels of PGF2,EP1 and NE.The plasm levels of prostaglandin (562.5±98.2 vs.663.2±119.0) pg/ml,epinephrine (24.9±4.8 vs.29.7±3.5) pg/ml and norepinephrine (408.3 ±47.2 vs.499.6±45.6) pg/ml in patients of group L were lower than those in group C (P<0.05)48 h after the surgery.The cytotoxicity of NK cell was higher in group L than that in group C (44.1 ±5.0 vs.37.1±5.5)% (P<0.05) 48 h after the surgery.The ratio of p-PKA/PKA was lower in group L than that in guoup C (0.060±0.008 vs.0.099±0.011) (P<0.05) at the end of the surgery.Conclusion Perioperative intravenous lidocaine infusion can reduce the level of plasma catecholamine and PGE2,and protect the cytotoxicity of NK cell,possibly via inhibiting of cAMP-PKA signaling pathway.

Article in Chinese | WPRIM | ID: wpr-606635


Objective To investigate the changes and clinical significance ot the expression ot microRNA in plasma and peripheral blood mononuclear cells in patients with schizophrenia.Methods 174 patients with schizophrenia in Danzhou People 's Hospital were selected as the case group and the other 80 healthy persons as control group The relative expression levels of 8 microRNA in two groups of plasma and pcripheral blood mononuclear cells were detected by real-time quantitative fluorescent PCR(MiR-195,MiR-346,MiR-181b,MiR-212,MiR-30e,MiR-432,MiR 7,MiR-34a),and the differences of microRNA in the plasma and peripheral blood mononuclear cells were compared between the two groups.ROC curve was used to analyze the sensitivity and specificity of microRNA as diagnostic criteria for schizophrenia and Logistic regression analysis of the relative risk of microRNA in schizophrenia.Results The expression levels of MiR-195,MiR-181b,MiR-132,MiR-30e,MiR-7 and in the patients of the case group(3.11±1.05,2.18±0.72,1.85±0.74 and 9.61±1.87) were significantly higher than those in the control group(4.48±1.07,2.92±0.86,3.53±1.07 and 11.96±2.73,P<0.05 or P<0.01).The expression levels of MiR-181b,MiR-212,MiR-30e and MiR-34a in peripheral blood mononuclear cells of patients in the case group (-4.20±1.16,0.27 ±0.55,-4.83± 1.05 and 2.64± 1.08) were significantly higher than those in control group (-3.56±0.81,0.91±0.68,-3.49±1.22 and 3.95±1.03,P<0.05 or P<0.01).Logistic regression analysis showed that plasma MiR-181b and MiR-30e were significantly relative risk (OR=2.357,95 % CI:1.361 ~ 4.093;OR=2.064,95 % CI:1.147~3.815),and peripheral blood mononuclear cells MiR-30e also had significant relative risk (OR=1.628,95 %CI:0.914~2.926).ROC curve analysis showed that 95%CI and AUC in plasma and peripheral blood mononuclear cells of MiR-181b were 0.702 (0.784~0.632),0.658 (0.593 to 0.736),and plasma and peripheral blood mononuclear cell of MiR-30e were 0.775 (0.706~0.857),0.758 (0.686~0.839),respectively.Spearman correlation analysis showed that plasma MiR 181b and plasma MiR-30e were significantly correlated (r=0.547,P =0.043).Conclusion Abnormal expression of microRNA in patients with schizophrenia,and plasma and peripheral blood mononuclear cells MiR-181b and MiR-30e had good diagnostic value for schizophrenia patients.

Article in Chinese | WPRIM | ID: wpr-443453


A total of 26 male patients with urethral condyloma acuminatum took Polystictus glycopeptide capsules orally for 4 weeks plus 1-5 sessions of photodynamic therapy.Prior to the first session,human papilloma virus (HPV) genotype of wart tissue was detected.Verrucous body fell off completely.The follow-up period was 6 months.There was one recurrent case.And 24 cases had HPV genotype DNA.The positive rate of high-risk infection was 46.2% ; the high/low-risk mixed type cross infection was present.The multiple HPV6 low-risk infection rate was 23.1% and multiple high-risk infection rate 58.3% ; highrisk type HPV16,HPV31,HPV58 and HPV6,HPV11 were all of mixed types.