Objective:To analyze the immunocontraceptive efficacy of CZP3 DNA vaccine by electric pulse stimulating and the correlation between antibody level and antifertility. Methods: The mice were immunized with 5, 10, 20 and 50 μg DNA vaccine pcDNA3. 0-CZP3 respectively and were stimulated at leg muscle by electric pulse. Detect the antibody level of each mice group and the highest antibody titer by ELISA. After mated with male mice for three weeks,calculated infertility rate of mice,the average litter size of each immune group and did statistical analysis. Results:Compared with the intramuscular injection alone,the electric pulse stimulation could increase serum antibody titers of mice significantly, the antibody titer of 50 μg group which was stimulated by electric pulse reached 1∶4 000. Moreover,the infertility of the mice increased and the mice average litter size reduced along with the increase of CZP3 antibody level significantly. Statistical analysis showed that the antibody level and the average litter size had a negative correlation. Conclusion:The electric pulse stimulation can improve the immune level and reduce the fertility efficiency of mice significantly,therefore establishes a foundation for further application of CZP3 DNA vaccine in stray dog contraception.

Objective To measure the lethal dosage values ( LD50 ) of i.v.asarone injection for mice and to establish a standard for abnormal toxicity test of asarone injection to potentially reduce the occurrence of adverse drug reaction.Methods To obtain the LD50 value, a weighted linear probit regression method ( Bliss method) is employed. The limit of abnormal toxicity test is determined according to Appendix XI C in its 2010 edition of the Chinese pharmacopoeia.Results It is found that the LD50 of intravenously asarone injection in mice ranges from 51.9 to 153.1 mg/kg.The abnormal toxicity test should be added as an additional item in the standard.Conclusions Based on analyses in this study, an appropriate limit of abnormal toxicity test is 15 mg/kg, which is also in line with current medical standard in China.

Objective:To explore the therapeutic effect of intra-articular injection of triptolide nanomaterials on rabbit antigen-induced knee arthritis.Methods:Twenty-seven New Zealand white rabbits were randomly divided into 4 groups. After antigen-induced arthritis (AIA) model were induced, the knee joints of triptolide nanomaterials (TPNA) group, triptolide (TP) group and betamethasone (BS) group were injected intra-articularly under ultrasound guidance with triptolide nanomaterials, triptolide and betamethasone respectively, 7 rabbits in each group. And the other 6 rabbits were punctured but not injected with any drugs as the control group. The pathological changes of joint swelling, synovitis and bone erosion were examined. Student's test, repeated measure data of analysis of variance (ANOVA), Mann-Whitney U test and Kruskal- Wallis test were used for statistical analysis. Results:① Before treatment, the knee joint diameters of the TPNA group, TP group, BS group and control group were (2.02±0.08) cm, (2.08±0.06) cm, (2.10±0.06) cm and (2.18±0.07) cm, respectively. After one week of administration, the knee joint diameters of the above groups were (1.85±0.06) cm, (1.89±0.07) cm, (1.93±0.08) cm and (2.15±0.08) cm, respectively. Knee joint swelling was significantly reduced in each treated group after a week of intra-articular injection. With the extension of treatment, the diameter of rabbit knee joints in each experimental group gradually decreased gradually ( F=58.83, P<0.01; F=53.78, P<0.01; F=68.24, P<0.01), and the diameter of rabbit knee joints in the TP group, TPNA group and BS group was significantly smaller than that of the control group ( F=63.83, P<0.01; F=71.94, P<0.01; F=140.79, P<0.01). ② The synovitis score of TP group was lower than that of the control group ( Z=-2.082, P<0.05), which was mainly mild synovitis. While the synovitis scores of TPNA group and BS group were lower than that of TP group ( Z=-2.082, P<0.05; Z=-2.687, P<0.05), most of which were free from synovitis. There was no statistical significant difference between BS group and TPNA group ( Z=-1.000, P>0.05). ③ The pathological scores of bone destruction in the TPNA group, TP group and BS group were all reduced compared with the control group ( Z=-2.505, P<0.05; Z=-2.216, P<0.05; Z=-2.505, P<0.05). There was no statistical significant difference between the TPNA group, TP group and BS group ( χ2=0.588, P>0.05). Conclusion:Intra-articular injection of triptolide nanomaterials can relieve joint swelling, reduce synovitis, and delay bone erosion. Its effect is similiar to glucocorticoid, better than simple triptolide. Triptolide nanomaterials have the potential to be an effective drug for arthritis by intra-articular injection.

Objective To test the effects of Vitamin D (VitD) on endothelial nitric oxide(NO) production and to study the signal pathway leading to NO release.Methods In vitro cultured human umbilical vein endothelial cells (HUVEC) were treated with various concentrations of VitD(0 mmol/L,0.01 mmol/L,0.10 mmol/L,1.00 mmol/L,10.00 mmol/L) for 60 min,and VitD at concentration of 1.00 mmol/L at different time points (30 min,60 min,90 min,120 min).The effect of VitD on NO production in presence of VitD receptor(VDR) agonist(ZK191784) or antagonist(ZK159222) for 60 min were examined in cell culture supernatant with kit for the detection of nitric oxide fluorescent probe(DAF-FM DA).HUVEC was cultured with VitD in presence of VDR agonist or antagonist for 60 min,and the effect of VitD on NO production with DAF-FM DA and the protein expression and phosphorylation of Caveolin-1 and endothelial nitric oxide synthase(eNOS) were detected by Western blot,respectively.Results VitD caused a concentration-dependent increase in NO production.The maximum effect was observed at a concentration of 1.0 mmol/L and the optimal time of stimulation was 60 min.Effects induced by VitD were enhanced by VDR agonist,and abolished by antagonist.VitD and VDR agonist maintained the expression of Caveolin-1 at the same low phosphorylation level the same as normal,increased the phosphorylated level of eNOS.However,VDR antagonist increased the phosphorylation of caveolin-l,but reduced the level of eNOS phosphorylation,respectively.Conclusions VitD can induce a significant increase in endothelial NO production through VDR.VitD interaction with VDR causes the low phosphorylation of caveolin-1 leading to eNOS activation and NO production.

A method for quantitative detection of florfenicol by colloidal gold lateral flow immunoassay was developed.The experimental conditions including pH value, concentrations of antibody in the process of conjugation between the colloidal gold and antibody, amount of gold-labeled antibody, concentration of the antigen sprayed on test lines (T line), and detection time were optimized.With a colloidal gold strip reader, the signal intensity of T lines and control lines (C line) on lateral flow strips was recorded.The T/C ratio of negative control and positive samples was defined as B0 and Bx, and the standard curve was established by plotting the Bx/B0 ratio against the concentration of florfenicol.This assay showed a good linear range from 0.1 to 1.5 ng/mL with the limit of detection of 0.08 ng/mL, while the result could be obtained within 15 min.The result showed that this quantitative method was convenient and rapid, and could be used in screening a large amount of samples on site.

Objective:To analysis the status of the mental health and related factors of fire fighters in Tianjin Binhai New District, and to provide suggestions for their psychological health protection.Methods:399 fire fighters in Tianjin Binhai New District were selected as study subjects in Jan to April 2019. Depression symptoms were measured by the depression module of the Patient Health Questionnaire (PHQ-9) . The Chinese version of Efrort Reword Imbalance (ERI) Questionnnaire were used to investigate and evaluate their occupational stress. Chi-Square test was used to analysis Categorical data. Binary logistic regression model was used to analysis the ralated factors of depression.Results:Among the 399 fire fighters, 71.1% (280/394) were found high level of depression symptom. The detection rates of depression symptoms in the related influceing factors ERI、station、disease、life pressure、eating habits and sleep disorder occupational stress were difierent ( P<0.05) . Sleep disorder, life pressure and ERI occupational stress were risk factors for depressive symptoms ( OR=1.921, 95% CI=1.002-3.682; OR=2.852, 95% CI=1.561-5.212; OR=2.367, 95% CI=1.163-4.818, P<0.05) . Conclusion:The rate of depression of fire fighters is relatively higher. Government should pay attention to and take measures to improve the psychological condition of fire fighters.

Objective:To analysis the status of the mental health and related factors of fire fighters in Tianjin Binhai New District, and to provide suggestions for their psychological health protection.Methods:399 fire fighters in Tianjin Binhai New District were selected as study subjects in Jan to April 2019. Depression symptoms were measured by the depression module of the Patient Health Questionnaire (PHQ-9) . The Chinese version of Efrort Reword Imbalance (ERI) Questionnnaire were used to investigate and evaluate their occupational stress. Chi-Square test was used to analysis Categorical data. Binary logistic regression model was used to analysis the ralated factors of depression.Results:Among the 399 fire fighters, 71.1% (280/394) were found high level of depression symptom. The detection rates of depression symptoms in the related influceing factors ERI、station、disease、life pressure、eating habits and sleep disorder occupational stress were difierent ( P<0.05) . Sleep disorder, life pressure and ERI occupational stress were risk factors for depressive symptoms ( OR=1.921, 95% CI=1.002-3.682; OR=2.852, 95% CI=1.561-5.212; OR=2.367, 95% CI=1.163-4.818, P<0.05) . Conclusion:The rate of depression of fire fighters is relatively higher. Government should pay attention to and take measures to improve the psychological condition of fire fighters.

OBJECTIVETo screen of high cariogenicity Streptococcus mutans (S. mutans) strains isolated from clinical specimens preliminary.

METHODSAcidogenicity, aciduricity, extracellular polysaccharide production and adhesion of 41 strains of S. mutans isolated from clinical specimens were investigated to screen high cariogenicity S. mutans strains.

RESULTSThere were different cariogenicity among 41 strains of S. mutans, in which 3 strains of S. mutans had all high ability to produce extracellular polysaccharide, adhere to the saliva-coated hydroxyapatite, produce acid and tolerate acid, indicated there were 3 strains with high cariogenicity S. mutans strains isolated from clinical specimens. Another 3 strains of S. mutans with all low ability to produce extracellular polysaccharide, adhere to the saliva-coated hydroxyapatite, produce acid and tolerate acid indicated they were low cariogenicity S. mutans strains isolated from clinical specimens.

CONCLUSIONWe may have obtained high cariogenicity S. mutans strains isolated from clinical specimens.

Dental Caries ; Durapatite ; Humans ; Saliva ; Streptococcus mutans

OBJECTIVETo identify Streptococcus mutans (S. mutans) strains from clinical samples.

METHODSPlaque samples from caries-active and caries-free sites on enamel surfaces were obtained and cultivated for S. mutans isolation. Morphology, biochemistry, automatic microorganism analysis system and polymerase chain reaction using primers homologous to surface protein antigen I/II (spaP), glucosyltransferase B (gtfB) and dextranase (dexA) were used to identify S. mutans. Genotype of isolated S. mutans was determined by arbitrarily primed polymerase chain reaction.

RESULTSForty-six strains of S. mutans were obtained from the 32 subjects and were identified as S. mutans by biochemistry, automatic microorganism analysis system and polymerase chain reaction. Five identical genotypes were found by arbitrarily primed polymerase chain reaction.

CONCLUSIONForty-one strains of S. mutans with different genotype were obtained from clinical samples.

Dental Caries ; Dental Plaque ; Genotype ; Glucosyltransferases ; Humans ; Polymerase Chain Reaction ; Streptococcus mutans

OBJECTIVETo select and identify ssDNA aptamers specific to Streptococcus mutans strains with different cariogenicity isolated from clinical specimens.

METHODSSubtractive SELEX technology targeting the whole intact cells was used to screen for ssDNA aptamers specific to the clinical isolates Streptococcus mutans strains with different cariogenicity. Radioactive isotope, flow cytometry, gene cloning and sequencing, MEME online software and RNA structure analysis software were employed to analyze the first and secondary structures of the aptamers and identify the screened aptamers.

RESULTSDetection by radioactive isotope showed sufficient pool enrichment after 9 rounds of subtractive SELEX. Flow cytometry showed that the selected aptamers H1, H16, H4, L1, L10 and H19 were capable of binding specifically with highly cariogenic Streptococcus mutans strains but not with strains with a low cariogenicity. The aptamer H19 had the strongest binding capacity to highly cariogenic Streptococcus mutans strains, with a dissociation constant of 69.45∓38.53 nmol/L.

CONCLUSIONWe have obtained the ssDNA aptamers specific to the clinical isolates of highly cariogenic Streptococcus mutans strains.

Aptamers, Nucleotide ; genetics ; Cloning, Molecular ; DNA Primers ; Dental Caries ; microbiology ; Gene Library ; Humans ; Nucleic Acid Conformation ; SELEX Aptamer Technique ; Species Specificity ; Streptococcus mutans ; classification ; genetics ; isolation & purification