1.Effect of retinoblastoma binding protein 4 (RBBP4)on Sp1-mediated transcription of HIV long terminal repeat in 293 T cells
Juan WANG ; Jin YANG ; Zongxing YANG ; Linfang CHENG ; Nanping WU
Chinese Journal of Clinical Infectious Diseases 2017;10(1):31-36
Objective To investigate the effect of retinoblastoma binding protein 4 (RBBP4)in Sp1 -mediated HIV long terminal repeat(LTR)transcription.Methods RBBP4 expression vector and Sp1 expression vector were respectively co-transfected into 293 T cells with HIV promoter pHIV-LTR-Luc or Sp1 site mutated pHIV-LTR-sp1 -mut by liposome transfection,and the transfected cells were examined by dual luciferase reporter assay system.The effect of RBBP4 on the binding of Sp1 to LTR was further studied by chromatin immunoprecipitation (ChIP)and electrophoretic mobility shift assay (EMSA).Results The relative firefly luciferase activity activated by Sp1 was decreased from 62.5 to 16 at the dose of 500 ng of RBBP4 expression vector (t =14.52,P <0.01 ).When the Sp1 binding sites were mutated,the effects of 100,300 or 500 ng of RBBP4 expression vector on the firefly luciferase activity of HIV LTR were not statistically significance (t =1 .897,2.357 and 3.162,all P <0.05).ChIP results showed that when the binding of RBBP4 on HIV LTR increased,the binding of Sp1 on HIV LTR increased significantly (t =11 .93,P <0.01 ),while the reduced binding of RBBP4 on HIV LTR significantly attenuated the binding of Sp1 onto LTR(t =11 .38,P <0.01 ).The effect of RBBP4 on the binding of Sp1 to DNA in ChIP assays was further verified by EMSA assays.Conclusion RBBP4 can inhibit the Sp1 -mediated HIV LTR transcription in 293 T cells.
2.Determination of 9 Phenoxy Acid Herbicides in Soil by Solid Phase Extraction and LC/MS/MS
Yanbin YU ; Li LI ; Jun YANG ; Pigong TAN ; Zongxing LIU
Chinese Journal of Analytical Chemistry 2014;(9):1354-1358
A method for the determination of 9 phenoxy acid herbicides in soil by solid phase extraction and LC/MS/MS with matrix correction was developed. The sample was treated by ultrasonic extraction twice with 1. 5% formic acid in acetonitrile. The extract was cleaned up with NH2 solid phase extraction column. A C18 column with formic acid in pure water/methanol mixture as the mobile phase was used under gradient elution. Mass spectral acquisition was done in negative ion mode by multiple reactions monitoring ( MRM ) . Two transitions per compound were used to provide qualitative and quantitative analysis. There were good linear relationships in the range of 2-250 μg/L for 9 phenoxy acid herbicides. The correlation coefficients were in the range of 0 . 9927-0 . 9972 . The method detection limits were in the range of 1 . 66-3 . 82 μg/kg . The average recoveries at three spiked levels ( 10 , 40 , 200 μg/kg ) ranged from 85 . 3% to 110 . 0% with the relative standard deviations (RSD) of 3. 2%-12%.
3.Study on anti-microbial activity of allicin on campylobacter jejuni
Hongtao PAN ; Zongxing YANG ; Wencui ZHAO ; Yanping WANG ; Haiyan SONG ; Qi ZHANG
Journal of Pharmaceutical Practice 2015;(2):147-149,170
Objective To study the anti‐microbial activity and strength of allicin on Campylobacter jejuni .Methods Disc diffusion method (K‐B) was used to determine the diameter of the bacteriostatic ring .The minimal inhibitory concentra‐tion (MIC) of allicin ,ciprofloxacin and erythromycin were detected by constant broth dilution method ,respectively .The mini‐mal inhibitory concentrations and bacteriostatic rate of allicin ,ciprofloxacin and erythromycin were compared .Results The anti‐microbial activity on Campylobacter jejuni of allicin (MIC 12 .8 μg/ml ,bactetiostatic rate 90 .40% ) was better than that of ciprofloxacin (MIC 20 .48 μg/ml ,bactetiostatic rate 90 .21% ) and erythromycin (MIC 35 .84 μg/ml ,bactetiostatic rate 90 . 33% ) .Conclusion Allicin has anti‐microbial activity on Campylobacter jejuni in vitro .
4.Incidence and risk factors of renal injury in human immunodeficiency virus infection/acquired immunodeficiency syndrome patients with poor immune reconstitution
Danyan YU ; Xumin YING ; Huili LI ; Zhangqing ZHOU ; Xiaofeng ZHENG ; Dingyan YAN ; Zongxing YANG ; Wenhui ZHANG ; Guoxiang ZHENG ; Zhongdong ZHANG ; Jianhua YU
Chinese Journal of Infectious Diseases 2023;41(8):502-506
Objective:To investigate the incidence and risk factors of renal injury in human immunodeficiency virus (HIV) infection/acquired immunodeficiency syndrome (AIDS) patients with poor immune reconstitution.Methods:The HIV infection/AIDS patients with poor immune reconstitution who were visited Second Department of Infection of Hangzhou Xixi Hospital from January to December 2021 were enrolled. The clinical data and laboratory examinations of the patients were collected, and the relevant risk factors were analyzed by logistic regression.Results:Among 303 HIV infection/AIDS patients with poor immune reconstitution, 59(19.5%) patients had renal injury. Logistic regression analysis showed that hypertension (odds ratio ( OR)=0.200, 95% confidence interval (95% CI) 0.065 to 0.618, P=0.005), taking tenofovir ( OR=0.275, 95% CI 0.130 to 0.580, P=0.001), hypoproteinemia ( OR=1.045, 95% CI 1.006 to 1.086, P=0.022), and low CD4 + T lymphocytes level ( OR=1.009, 95% CI 1.003 to 1.014, P=0.001) were risk factors for renal injury. Conclusions:The incidence of renal injury in HIV infection/AIDS patients with poor immune reconstitution is high. Hypertension, taking tenofovir, hypoproteinemia, and low CD4 + T lymphocytes level are risk factors for renal injury in patients.
5.Periodic revalidation of autoverification for blood analysis and its suitability evaluation of application
Yingtong LI ; Xuejun WANG ; Wei XU ; Linlin QU ; Xianqiu CHEN ; Lijing WEI ; Ying WANG ; Hongli SHAN ; Zongxing YANG ; Yue CAI ; Xiaoquan YANG ; Wenrui SUN ; Dan LI ; Yue ZHANG ; Xi WANG ; Jin LIANG ; Jing HUANG ; Jiancheng XU ; Haiyan WANG ; Fang LIU ; Weining JIANG ; Chengming SHANG
Chinese Journal of Laboratory Medicine 2020;43(10):1021-1031
Objective:To conduct periodic revalidation of the 15 items and 43 terms autoverification rules of blood analysis after 1 year of application, analyze the application suitability and make the rules improved.Methods:Track the results of 528 010 blood analysis samples of our hospital from August 1, 2019 to January 31, 2020, and analyze the pass rate and interception rate of autoverification; 600 specimens in total were selected randomly for microscope examination, including 300 specimens which touched autoverification rules (1 012 items of autoverification rules) and were intercepted by autoverification and 300 specimens which untouched autoverification rules and were released by autoverification. The abnormal characteristics and unacceptable Delta check of the specimens also need to be concerned at the same time.The false negative rate and false positive rate, true negative rate, true positive rate and pass correct rate of autoverification were verified and compared with the rate of the second phase verification when the autoverification rule was established. The false negative rate, false positive rate, true negative rate and true positive rate of the Delta check rule which 54 716 specimens touched were calculated and compared with the second phase verification rate when the autoverification rule was established.The results of microscopic examination were used as the gold standard for the calculation of the rates, and P<0.05 was considered as a significant difference. The false positive and true positive of 1 012 autoverification rules were analyzed item by item.The false positive and true positive of 108 specimens which touched blast cell autoverification rule were analyzed terms by terms. The mean TAT and median TAT of 528 010 specimens and 193 750 outpatient specimens were calculated respectively, and the report percentages of 528 010 samples that TAT<30, 30-60 and>60 min were calculated respectively. Analyze and evaluate the application suitability of autoverification rules to juge whether they meet the needs of doctors and laboratory. The design process and the rules and application process of autoverification were optimized and improved.Results:The autoverification pass rate was 63.06% (332 971/528 010), the interception rate was 36.94% (195 039/528 010). The false negative rate was 1.00% (1/600), the false positive rate was 12.67% (76/600), the true negative rate was 49% (294/600), the true positive rate was 37.33% (224/600), and the correct rate was 98% (294/300). The pass rate, true negative rate, true positive rate and correct rate of the periodic reverification group were higher than the second phase verification group, the false negative rate and false positive rate were lower than that the second phase verification group. The false negative rate and true positive rate of the Delta check of periodic verification group were lower than that the second phase verification group, the false positive rate and true negative rate were higher than the second phase verification group, there were significant differences in the comparition results. The mean TAT of 528 010 specimens was25 min, and the median TAT was 22 min. The mean TAT of 193 750 outpatient specimens was 23 min, and the median TAT was 20 min. The report percentages of 528 010 samples that TAT<30 min, 30 min-60 min and>60 min were 83.30% (439 819/528 010), 8.00% (42 250/528 010) and 8.70% (45 941/528 010), respectively.Conclusion:The results of periodic revalidation of autoverification after 1 years application show that the 15 items and 43 terms autoverification rules of blood analysis could meet requirements about the accuracy and efficiency of the laboratory, and have a good suitability for application.