OBJECTIVE:To establish a RP-HPLC method for the determination of aristolochic acid A in decoction of Caulis Aristolochiae Manshuriensis METHODS:The analytical column was Spherisorb ODS2 column(4 6mm?250mm,5?m) The mobile phase consisted of a mixture,methanol-water-acetic acid(70∶27∶1) The flow rate was 1 0ml/min The UV detection wavelength was 250nm RESULTS:The linear range was 0 0 128?g～0 4 096?g(r=0 9 999) The regression equation was Y=4 553 7+5 388 319 3X The average recovery of aristolochic acid A was 97 33%(RSD=2 34%) CONCLUSION:This method is simple,sensitive and accurate

Objective:To explore the characteristics of type 3 secretion system and biofilm of Pseudomonas aeruginosa in diabetic foot wound, and to analyze the relationship between these factors, as well as to the antibiotic sensitivity.Methods:Thirty-three strains of Pseudomonas aeruginosa were collected from the foot wounds of diabetic foot inpatients in Tianjin Medical University Chu Hsien-I Memorial Hospital from February 1, 2018 to December 31, 2018. Thirteen strains of Pseudomonas aeruginosa were collected from non-diabetic wounds. All strains were tested for antibiotic sensitivity. The virulence genes exoS or exoU of Pseudomonas aeruginosa and the ability of biofilm formation were tested. The characteristics of exoS or exoU and biofilm of Pseudomonas aeruginosa were analyzed. Patients′ clinical outcomes were also analyzed.Results:Pseudomonas aeruginosa with exoS gene was the major pathogen, 90.9% found in diabetic foot group and 84.6% in control group, with no significant difference( χ2=0.54, P=0.46). The drug-resistant strains of Pseudomonas aeruginosa with exoS accounted for 16.7% in diabetic foot group and 18.2% in control group, also with no significant difference( χ2=0.18, P=0.83). There were 5 strains of Pseudomonas aeruginosa carrying exoU, 3 strains in diabetic foot group, of which 1 was resistant, 2 in control group, no resistant strain. Pseudomonas aeruginosa increased the ability of biofilm formation in diabetic foot group, accounting for 57.6%, and for resistant strains, 83.3% of them increased the biofilm formation ability. Two kinds of Pseudomonas aeruginosa produced different biofilms, but they were effectiveless for carbapenem antibiotics. The times of debridement ( P<0.01), time of antibiotic use ( P<0.01) were more in biofilm wound, but the healing rate reached 75%-90%. Conclusion:Pseudomonas aeruginosa secreting ExoS is the main one in the diabetic foot wound. The ability of Pseudomonas aeruginosa to produce biofilm in DF wound is increased. Biofilm is one reason for its antibiotic resistance. Multiple debridement combined with sensitive antibiotics is an effective method to remove biofilm.

Objective To detect the expression of costimulatory molecules B7-H4 and programmed death ligand 1 (PD-L1) in bladder cancer,and to explore the correlation between them and clinicopathological features of bladder cancer.Methods Immunohistochemical staining was used to detect the expression of B7-H4 and PD-L1 in 98 cases of bladder urothelial carcinoma,which were confirmed by pathology from August 2014 to December 2015 in our hospital.There were 23 females,aged 45-82 years,with an average age of 67.8 years.Among them,42 cases of adjacent normal tissues were used as controls.The clinical stage,histological grade and recurrence of bladder cancer were collected,and the correlation between them was analyzed.Results The positive rates of B7-H4 and PD-L1 in bladder urothelial carcinoma were 54.1％ (53/98) and 59.2％ (58/98),respectively,and there was no expression in normal bladder tissues (P ＜ 0.05).The positive expression rate of B7-H4 in muscle invasive bladder cancer (MIBC) patients was higher than that in non-muscle invasive bladder cancer (NMIBC) [73.5％ (25/34) vs.43.8％ (28/64),P =0.005].The positive expression rate of B7-H4 in high-grade patients was higher than that of low-grade [70.0％ (21/30) vs.47.1％ (32/68),P =0.036].The expression rate of B7-H4 in high-risk group was higher than that of low-intermediate risk group [57.1％ (20/35) vs.27.6％ (8/29),P =0.018].The positive expression rate of PD-L1 in patients with MIBC was higher than that in NMIBC [79.4％ (27/34) vs.48.4％ (31/64),P =0.003].The PD-L1 expression rate of histological high-level group was higher than that of low-level group [73.3％ (22/30) vs.52.9％ (36/68)],but the difference was not statistically significant (P =0.058).The PD-L1 expression rate in high-risk group was 68.6％ (24/35),and also higher than low-middle group 24.1％ (7/29) (P ＜ 0.05).There was a positive correlation between the expression of B7-H4 and PD-L1 in bladder urothelial carcinoma (r =0.318,P=0.002).The combined recurrence rate of the two groups was significantly higher than that of the negative expression of the two groups [66.7％ (14/21) vs.30.8％ (8/26),P=0.014].Conclusions The expression of B7-H4 and PD-L1 is up-regulated in bladder urothelial carcinoma,which is closely related to the clinical stage,histological grade,risk classification and recurrence of NMIBC.