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 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.

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.

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.

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

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

Objective To investigate the effect of prior statin use on outcome after intravenous thrombolysis in patients with acute ischemic stroke. Methods Consecutive patients with acute ischemic stroke treated with intravenous thrombolysis at the Department of Neurology, the Third People's Hospital of Chengdu from July 2014 to August 2017 were enrolled, and divided into the statin use group and nonstatin use group according to prior statin use. Symptomatic intracranial hemorrhage and the outcome at 90 days after onset (good outcome and poor outcome were defined as the modified Rankin Scale score 0-2 and > 2, respectively) in the two groups were compared, and multivariate logistic regression analysis was used to identify the effect of prior statin use on the outcome. Results A total of 327 patients were enrolled, including 68 (20. 80％ ) in the statin use group, and 59 (79. 20％ ) in the nonstatin use group. There were no significant differences in the incidence symptomatic intracranial hemorrhage (7. 35％ vs. 10. 04％; χ2 = 0. 453, P = 0. 501), good outcome rate at 90 days (69. 12％ vs. 66. 02％; χ2 = 0. 232, P = 0. 630), and mortality rate (7. 35％ vs. 7. 34％; P = 1. 000) between the statin use group and the nonstatin use group. Multivariable logistic regression analysis showed that prior statin use were not an independent risk factor for symptomatic intracranial hemorrhage (odds ratio 0. 658, 95％ confidence interval 0. 233-1. 857; P = 0. 429) and poor outcome at 90 dafter onset (odds ratio 0. 848, 95％ confidence interval 0. 424-1. 696; P = 0. 641) in patients treated with intravenous thrombolysis. Conclusion Prior statin use is not associated with symptomatic intracranial hemorrhage and outcome after intravenous thrombolysis in patients with acute ischemic stroke.

Objective:To compare the clinical efficacy of transperitoneal robot assisted laparoscopic radical prostatectomy (RARP) versus extraperitoneal single port RARP.Methods:A retrospective analysis was perfoumed on 142 cases of RARP from July 2019 to June 2020 in Robotic Minimally Invasive Center of Sichuan Provincial People's Hospital, including 82 cases(Group A) , aged 70.0(65.6-78.0), undergoing transperitoneal RARP and 60 cases(Group B), aged 68.2 years old(60.1-79.2), undergoing extraperitoneal single port RARP. For group A, PSA was 12.9(5.6-64.0) ng/ml, with 26 cases of less than 10 ng/ml(31.7%), 40 cases of 10-20 ng/ml(48.8%), and 16 cases of more than 20 ng/ml(19.5%), the Gleason score was 7.2(6.0-10.0), with 14 cases(17.1%) of ≤6, 56 cases(68.3%) of 7, and 12 cases(14.6%) of ≥8, and the prostate volume was 61.3(29.0-112.0) ml. There were 49 cases with BMI≤25 kg/m 2, accounting for 59.8%, and 33 cases with BMI>25 kg/m 2, accounting for 40.2%. There were 17 cases(20.7%) of T 1, 44 cases(53.7%) of T 2 and 21 cases(25.6%) of T 3. The proportion of lymph node dissection was 17.1%, and 4 cases(4.9%)had a history of operation. For group B, the PSA was 12.2(1.0-42.6)ng/ml, with 20 cases (33.3%) of <10 ng/ml, 31 cases(51.7%)of 10-20 ng/ml, and 9 cases (15%) of >20 ng/ml. Gleason score was 7.1(6.0-9.0), with 12 cases (20.0%) of ≤6, 42 cases (70.0%) of 7, and 6 cases (10.0%)of ≥8. Prostate volume was 42.4(31.2-72.8)ml on average. There were 37 cases (61.7%) with BMI≤25 kg/m 2, and 23 cases (38.3%)with BMI >25 kg/m 2 . There were 17 cases(28.3%)of T 1, 32 cases(53.3%)of T 2 and 11 cases(18.3%)of T 3.The proportion of lymph node dissection was 11.7% and 4 cases (6.7%) had a history of operation.There was no statistically significant difference in term of age, PSA level, Gleason score, BMI, clinical stage, proportion of lymph node dissection or history of operation between the two groups( P>0.05), but there was statistically significant difference for prostate volume( P<0.05). All operations were performed by the same operator. Four different ways of bladder neck and urethral dissociation was selected according to the intraoperative conditions in Group A, include VIP style, T-shape incision style, VIP plus T-shape incision style or the style along the lateral side of the bladder neck. Small and single anterograde incision stripping of bladder neck was routinely performed in the Group B. Postoperative follow-up was performed to compare the operation time, intraoperative blood loss, bladder neck and urethral anastomosis time, postoperative hospital stay, postoperative exhaust time, postoperative complications, positive rate of surgical margin, indwelling time of urinary catheter, urinary continence satisfaction rate of immediately after operation, 3 months and 6 months after operation, wound healing and aesthetics. Results:All of the operations were successfully completed under robot-assisted laparoscopy, and there was no conversion to open surgery. The operation time was 56.0(45.0-112.0) min in the Group A and 65.4(55.5-96.8) min in the Group B, and there was no statistically significant difference( P>0.05). The intraoperative blood loss was 76.2(30.0-120.5) ml and 55.6(45.5-114.6) ml, respectively, and the difference was not statistically significant( P>0.05). The time of bladder neck urethral anastomosis was 18.9(12.6-25.6) min and 16.2(10.7-19.3) min, respectively, and the difference was not statistically significant( P>0.05). The postoperative hospital stay days were 9.3(8.0-16.0) d and 8.4(7.0-13.0) d, respectively, and the difference was not statistically significant( P>0.05). The postoperative exhaust time was 1.3(0.7-3.0) d and 3.4(2.0-7.0) d, respectively, and the difference was statistically significant( P<0.05). There was 1 case of anastomotic fistula with ureteral injury in Group A, and no serious complication in Group B, and the difference was not statistically significant( P>0.05). The number of positive surgical margin in the two groups was 13(15.9%)and 9(15.0%)respectively, and the difference was not statistically significant( P>0.05). The indwelling time of urinary catheter after operation was 9(7-21) d and 6(4-8) d, respectively, and the difference was statistically significant( P<0.05). The number of patients with satisfactory urinary continence immediately after surgery, 3 months and 6 months after surgery in the two groups were 8(9.8%), 51(62.2%), 62(75.6%) and 17(28.3%), 43(71.7%) and 54(90.0%), respectively. The differences were statistically significant( P<0.05). The total incision lengths in the two groups were 12.1(10.4-13.4) cm and 5.6(5.0-6.0) cm, respectively, and the difference was statistically significant( P<0.05). Conclusions:The extraperitoneal single port RARP is safe and feasible, and the postoperative effect is similar to that of transperitoneal RARP. It has the advantages of shorter recovery time, higher urinary continence satisfaction rate, neater and more beautiful incision. The long-term therapeutic effect needs further confirming by prospective study.