Performance of yeast-like cell counting (YLCC) using the Sysmex UF-1000i for clinical candiduria screening.

Candiduria is common in clinical practice. However, an effective and convenient assay to screen for candiduria is still needed. This study aimed to evaluate the performance of the Sysmex UF-1000i urine analyzer for yeast-like cell counting (YLCC) to screen for candiduria prior to urine culture. We retrospectively analyzed data from 5233 urine samples from 1813 patients, including 837 males and 976 females. Urine culture and urinalysis-obtained YLCC data were used to estimate the performance of YLCC in diagnosing candiduria. Different cutoff values were used to calculate sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV). The YLCC-positive rates differed according to the Candida colony-forming units (CFU) counts in the urine samples. A sharp drop in YLCC-positive rate (from 64.3 to 22.0%) was observed between the urine groups with 104 CFUs and 103 CFUs. A cutoff value of 0 YLCs/µL results in the highest Youden index (0.71) with 77.04% sensitivity and 93.68% specificity. In a group of 34 hospitalized candiduria patients with serial urinalysis data, 25 were YLCC-positive before urine culture. In conclusion, YLCC with the Sysmax UF-1000i could serve as an auxiliary technique to exclude culture-negative specimens prior to urine culture. Positive YLCC results could imply candiduria, especially when persistent YLCC-positive results were observed.

Therapeutical effect of Allicin for colitis mice induced by DSS and its possible mechanisms / 中国免疫学杂志

Objective:To analyze the therapeutical effect of Allicin for colitis mice induced by DSS and its possible mechanisms. Methods:A total of 24 male mice were randomly divided into Control group,DSS group(2. 5% DSS,7 days) and Allicin group [DSS treatment and Allicin,10 mg/(kg·d),7 d]. Disease activity index,inflammatory score,TUNEL and Western bolt were performed to analyze the effect of Allicin on colitis induced by DSS. Results: With DSS group, the Allicin group had lower disease activity score and inflammatory score(P<0. 05). Allicin group had a lower number of intestinal epithelial cell apoptosis than that of DSS group,the difference was statistically significant(P<0. 05). Western bolt analysis shown that Allicin treatment significantly depressed the expression of p-JAK2 and p-STAT3 in intestinal mucosa when compared with these of DSS group ( P<0. 05 ) . Conclusion:Conclusion Allicin has significantly therapeutic effect on mice colitis induced by DSS, the possible mechanisms including anti-inflammatory effects,protected of the intestinal epithelial cells and inhibition of the JAK2/STAT3 signaling pathways.

Coincidence rates between empiric antimicrobial use and antimicrobial sus-ceptibility testing results in patients with pyelonephritis / 中国感染控制杂志

Objective To understand the empiric antimicrobial use in patients with pyelonephritis in a hospital,and pro-vide reference for clinical rational antimicrobial use.Methods Data of 620 patients with pyelonephritis admitted to the nephrology department of a hospital between January 2011 and September 2014 were collected,application of antimicrobial agents,coincidence between empiric antimicrobial use and antimicrobial susceptibility testing results in patients with different diseases and different ages were analyzed.Results Before antimicrobial susceptibility testing results were reported,620 pa-tients use 625 times of antimicrobial agents,5 of whom used two kinds of antimicrobial agents at the same time,8 varieties in 15 types of antimicrobial agents were involved,the most frequently used antimicrobial agents were third generation ceph-alosporins,cephamycins,and fluoroquinolones.The overall,partial,and non-coincidence rate between antimicrobial use and antimicrobial susceptibility testing results were 64.32%(n=402),8.32%(n=52),and 27.36%(n=171)respective-ly.The overall coincidence rate in patients with acute pyelonephritis was higher than those with chronic pyelonephritis (77.61% [n=357]vs 58.79%[n=97],P <0.05).The overall coincidence rate in patients <50 years old and ≥50 years old were 68.12%(156/229)and 75.25%(298/396)respectively,there was no significant different between two groups (χ2 =2.93,P =0.09).Conclusion The non-coincidence rate between empiric antimicrobial use and antimicrobial suscepti-bility testing results is high,measures needs to be taken to improve the empiric antimicrobials use.

Gene cloning and expression of the Tp0453 antigen immuno-dominant epitope fragment of Treponema pallidum and its potential use in serodiagnosis of syphilis / 中华传染病杂志

Objective To clone and express the Tp0453 antigen immuno-dominant epitope fragment of Treponema pallidum (Tp) in Escherichia coli,in an effort to develop serological tests with increased specificity for the diagnosis of syphilis.Methods The gene encoding Tp0453 recombinant outer membrane protein fragment was amplified by polymerase chain reaction (PCR),and inserted into expression vector pQE30 after T-A cloning,then confirmed by restriction map.The constructed recombinant plasmid pQE30-Tp0453 was transformed to E.coli M15 for expression induced by isopropyl β-D-1-thiogalactopyranoside.The expressed product was identified by Western blot,and purified by Ni2+-NTA agarose column chromatography.A double antigen sandwich enzymelinked immunosorbent assays (ELISA) was established by using the recombinant Tp0453 protein to test sera from 48 patients with positive Treponema pallidum particle agglutination test (TPPA),and 40 negative sera as control.Results The PCR amplicon of the target gene was about 490 bp.The recombinant plasmid pQE30-Tp0453 was correctly constructed and successfully expressed in E.coli M15.The expressed product,with a relative molecular of about 21 000,existed in a form of inclusion body,accounting for about 18％ of total somatic protein,and reached a purity of more than 95％ after purification.Western blot showed specific reaction of the expressed protein with Tp positive serum.The ELISA tests with the 88 clinical samples yielded a sensitivity of 97.9％ (47/48),and specificity of 100.0 ％ (40/40).The consistency of results between the ELISA test and the TPPA test was 98.9 ％ (87/88).Conclusion The expressed Tp0453 fragment has showed good immunoreactivity with serum from patients with syphilis,providing the foundation of further development of serological diagnostic kit with increased specificity for the diagnosis of TP infection.

Recombination and expression of a component of urease B subunit transmembrane protein of helicobacter pylori / 免疫学杂志

Objective　To recombine and express a component of urease B subunit transmembrane protein of helicobacter pylori. Methods　A 732 bp gene fragment of urease B subunit of helicobacter pylori was cloned into pET11C and transformed into BL21（DE3）E.coli. The positive clone was induced with IPTG. The expression of target protein was analysed by SDS-PAGE and Western blot. Results　It is successful to construct the recombinant plasmid pET-UreB0.7 containing urease B subunit 0.7 kb gene fragment. A protein （MW≈28 000 u） with immunoreactivity， was expressed by 19.8％ in BL21（DE3）E.coli induced with IPTG. Conclusions　The recombinant component of urease B subunit transmembrane protein may play a role in the research of its biological function and might be used as the vaccine against helicobacter pylori.