1.Molecular mechanisms of quinolone resistance in non-typhoidal Salmonella
Jiajun LUO ; Shihan ZENG ; Yulan HUANG ; Yulian FENG ; Fangyin ZENG ; Xiaoyan LI
Chinese Journal of Preventive Medicine 2024;58(2):248-253
By conducting retrospective analysis, this study aim to investigate the resistance mechanism of quinolones in non-typhoidal Salmonella (NTS). A total of 105 strains of NTS isolated from clinical specimens from the Fifth Affiliated Hospital of Southern Medical University from May 2020 to February 2021 were used as research objects. VITEK2 Compact automatic identification drug sensitivity analysis system and serological test were used to identify the strains. The sensitivity of the strains to ciprofloxacin, levofloxacin and nalidixic acid was detected by AGAR dilution method. The whole genome of 105 strains of NTS was sequenced. Abricate and other softwares were used to analyze drug-resistant genes, including plasmid-mediated quinolone resistance gene (PMQR) and Quinolone resistance determination region (QRDR). Serotypes and ST types were analyzed using SISTR and MLST, and phylogenetic trees were constructed. The results showed that the NTS isolated in this region were mainly ST34 Salmonella typhimurium (53.3%). The drug sensitivity results showed that the drug resistance rates of NTS to ciprofloxacin, levofloxacin and nalidixic acid were 30.4%, 1.9% and 22.0%, respectively, and the intermediate rates of ciprofloxacin and levofloxacin were 27.6% and 54.2%.A total of 46 (74.2%) of the 62 quinolone non-susceptible strains carried the PMQR gene, mainly qnrS1 (80.4%), followed by aac(6′)-Ib-cr(15.2%); there were 14 NTS and 8 NTS had gyrA and parC gene mutations, respectively. The gyrA was mutations at the amino acid position 87, Asp87Tyr, Asp87Asn, Asp87Gly, and Thr57Ser mutations were detected in parC. In conclusion, this study found that NTS had relatively high resistance to quinolones, carrying qnrS1 gene mainly resulted in decreased sensitivity of NTS to ciprofloxacin and levofloxacin, and gyrA:87 mutation mainly resulted in NTS resistance to Nalidixic acid; Salmonella typhimurium in clinical isolates showed clonal transmission and required further epidemiological surveillance.
2.Molecular mechanisms of quinolone resistance in non-typhoidal Salmonella
Jiajun LUO ; Shihan ZENG ; Yulan HUANG ; Yulian FENG ; Fangyin ZENG ; Xiaoyan LI
Chinese Journal of Preventive Medicine 2024;58(2):248-253
By conducting retrospective analysis, this study aim to investigate the resistance mechanism of quinolones in non-typhoidal Salmonella (NTS). A total of 105 strains of NTS isolated from clinical specimens from the Fifth Affiliated Hospital of Southern Medical University from May 2020 to February 2021 were used as research objects. VITEK2 Compact automatic identification drug sensitivity analysis system and serological test were used to identify the strains. The sensitivity of the strains to ciprofloxacin, levofloxacin and nalidixic acid was detected by AGAR dilution method. The whole genome of 105 strains of NTS was sequenced. Abricate and other softwares were used to analyze drug-resistant genes, including plasmid-mediated quinolone resistance gene (PMQR) and Quinolone resistance determination region (QRDR). Serotypes and ST types were analyzed using SISTR and MLST, and phylogenetic trees were constructed. The results showed that the NTS isolated in this region were mainly ST34 Salmonella typhimurium (53.3%). The drug sensitivity results showed that the drug resistance rates of NTS to ciprofloxacin, levofloxacin and nalidixic acid were 30.4%, 1.9% and 22.0%, respectively, and the intermediate rates of ciprofloxacin and levofloxacin were 27.6% and 54.2%.A total of 46 (74.2%) of the 62 quinolone non-susceptible strains carried the PMQR gene, mainly qnrS1 (80.4%), followed by aac(6′)-Ib-cr(15.2%); there were 14 NTS and 8 NTS had gyrA and parC gene mutations, respectively. The gyrA was mutations at the amino acid position 87, Asp87Tyr, Asp87Asn, Asp87Gly, and Thr57Ser mutations were detected in parC. In conclusion, this study found that NTS had relatively high resistance to quinolones, carrying qnrS1 gene mainly resulted in decreased sensitivity of NTS to ciprofloxacin and levofloxacin, and gyrA:87 mutation mainly resulted in NTS resistance to Nalidixic acid; Salmonella typhimurium in clinical isolates showed clonal transmission and required further epidemiological surveillance.
3.Cloning and function analysis of elongase of very long chain fatty acid gene Bmelo424 in silkworm.
Weidong ZUO ; Ning KANG ; Chunlin LI ; Yue LUAN ; Xiaoling TONG ; Fangyin DAI ; Cheng LU
Chinese Journal of Biotechnology 2019;35(3):435-444
Genes belonging to the elongases of very long chain fatty acid (ELOVL) family affect many physiological functions in organism. In this paper, Bmelo424 gene, a member of the ELOVL family in silkworm, was cloned and its ORF was 558 bp. Its protein sequence was predicted to have four transmembrane domains, six serine phosphorylation sites, eight threonine phosphorylation sites and four tyrosine phosphorylation sites, and its subcellular localization was in the endoplasmic reticulum. Secondary structure analysis showed that the percentage of alpha-helix and beta-strand was 26.7% and 20% respectively. The results of fluorescence quantitative PCR showed that Bmelo424 gene was expressed in all tissues of silkworm, especially with the highest expression in head. By heterologous expression of Bmelo424 gene in Saccharomyces cerevisiae, the effect of Bmelo424 gene on fatty acid elongation was studied. GC-MS results indicated that the fatty acid content of C16:1n-7 in S. cerevisiae with pYES2-Bmelo424 recombinant plasmid increased significantly, whereas the content of C16:0, C18:0 and C18:1n-9 decreased. The results of temperature stress revealed that Bmelo424 gene could improve the low temperature adaptability of S. cerevisiae, but its high temperature adaptability decreased. This provides a reference for exploring the function of Bmelo424 gene in silkworm.
Acetyltransferases
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Amino Acid Sequence
;
Animals
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Bombyx
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Cloning, Molecular
;
Fatty Acids
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Saccharomyces cerevisiae
4.Performance evaluation of the hook effect of five immunoturbidimetric kits for the detection of specific proteins
Jie YANG ; Ling LI ; Hanyun ZHANG ; Peng ZHANG ; Fangyin ZENG
International Journal of Laboratory Medicine 2019;40(2):140-143
Objective To evaluate the performance of hook effect of five immunoturbidimetric kits for the detection of specific proteins on biochemical analyzers.Methods Five immunoturbidimetric kits with higher market share that came from Beijing BSBE (A), Sichuan maccura (B), Shenzhen Mindray (C), Ningbo Medical System (D) and Beijing Leadman (E) were used to determine six specific proteins.A series of concentration gradient samples were prepared and tested to compare the performance of hook effect from different manufactures′kits when the analytical measurement ranges were known.Results In the five kits, the upper limits of the safe range of antigen excess about ASO, hs-CRP andβ2-MG were relatively higher in B and C.No hook effect occurred at the approximate concentration of 10 000IU/mL, 1 000mg/L and 226mg/L respectively.The highest upper limits for CysC were C and E kits, and both were greater than 112mg/L.The upper limits of the safety range for other manufacturers were more than 700mg/L about RBP except for D.The maximum upper limit of mALB was D.Hook effect did not appear at the concentration of 43 560mg/L approximately.Conclusion Different manufactures′immunoturbidimetric kits have different hook effect performance.The laboratories should verify the hook effect performance before using the kits, and select the most suitable kit to prevent hook effect.
5.Efficacy and safety of Gemcitabine combined with cisplatin in the treatment of bladder cancer
Zongping WANG ; Yipeng XU ; Fangyin LI ; Shaoxing ZHU
Chinese Journal of Biochemical Pharmaceutics 2017;37(5):316-317,320
Objective To evaluate the clinical efficacy and safety of gemcitabine combined with cisplatin in the treatment of bladder cancer.MethodsFrom October 2012 to August 2015, 92 patients with bladder cancer were enrolled in our hospital.Patients were divided into observation group and control group by random number table method.On the basis of routine nutrition support and symptomatic treatment, cisplatin was administered by intravenous infusion of cisplatin 70mg/m2 in the first 3d in control group.On the 1d and 8d, gemcitabine 1000mg/m2 was intravenously infused in observation group and 21 days treatment was taken continuous for 2 courses.Curative effect, IL-17, IL-18, transforming growth factor-β (TGF-β), vascular endothelial growth factor (VEGF) and urinary TGF-β1 levels and adverse reactions of two groups were comparatively studied.ResultsThe total effective rate in the control group (63.05%) was significantly lower than that in the observation group (76.09%) (P<0.05).After treatment, the levels of serum IL-17, IL-18, TGF-β1 and VEGF in the two groups were significantly decreased and with significant difference between two groups (P<0.05).The levels of urinary TGF-β1 were significantly increased and with significant difference between two groups (P<0.05).The incidence of adverse reactions in the control and observation groups was 15.22% and 6.52%, respectively.There was no significant difference between the two groups.ConclusionGemcitabine combined with cisplatin has a significant clinical efficacy and safety in the treatment of bladder cancer.
6.Urinary metabolomics study of renal cell carcinoma based on gas chromatography-mass spectrometry.
Lin ZHANG ; Ling LI ; Hairui KONG ; Fangyin ZENG
Journal of Southern Medical University 2015;35(5):763-766
OBJECTIVETo identify the biomarkers of renal cell cancer (RCC) through urine metabolic analysis.
METHODSUrine samples of 27 RCC patients, 26 patients with other urinary cancers and 26 healthy volunteers were examined with gas chromatography-mass spectrometry (GC-MS). SIMCA-P+12.0.1.0 software was used for principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA) to screen for the differential metabolites.
RESULTSPCA (R2X=0.846, Q2=0.575) and OPLS-DA (R2X=0.736, R2Y=0.974, Q2Y=0.897) model were established for the RCC patients and control subjects. Fourteen metabolites were selected as the characteristic metabolites, including pentanoic acid, malonic acid, glutaric acid, adipic acid, amino quinoline, quinoline, indole acetic acid, and tryptophan, whose levels in the urine were significantly higher in the RCC patients than in the normal subjects (P<0.01); the RCC patients showed significantly higher urine contents of pentanoic acid, phenylalanine, and 6-methoxy-nitro quinoline than those with other urinary tumors (P<0.01).
CONCLUSIONThe urine metabolites identified based on GC-MS analysis can distinguish RCC patients from patients with other urinary cancers and healthy subjects, suggesting their potential as diagnostic markers for RCC.
Biomarkers ; urine ; Carcinoma, Renal Cell ; urine ; Discriminant Analysis ; Gas Chromatography-Mass Spectrometry ; Humans ; Least-Squares Analysis ; Metabolome ; Metabolomics ; Principal Component Analysis ; Software
7.Urinary metabolomics study of renal cell carcinoma based on gas chromatography-mass spectrometry
Lin ZHANG ; Ling LI ; Hairui KONG ; Fangyin ZENG
Journal of Southern Medical University 2015;(5):763-766
Objective To identify the biomarkers of renal cell cancer (RCC) through urine metabolic analysis. Methods Urine samples of 27 RCC patients, 26 patients with other urinary cancers and 26 healthy volunteers were examined with gas chromatography-mass spectrometry (GC-MS). SIMCA-P+12.0.1.0 software was used for principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA) to screen for the differential metabolites. Results PCA (R2X=0.846, Q2=0.575) and OPLS-DA (R2X=0.736, R2Y=0.974, Q2Y=0.897) model were established for the RCC patients and control subjects. Fourteen metabolites were selected as the characteristic metabolites, including pentanoic acid, malonic acid, glutaric acid, adipic acid, amino quinoline, quinoline, indole acetic acid, and tryptophan, whose levels in the urine were significantly higher in the RCC patients than in the normal subjects (P<0.01); the RCC patients showed significantly higher urine contents of pentanoic acid, phenylalanine, and 6-methoxy-nitro quinoline than those with other urinary tumors (P<0.01). Conclusion The urine metabolites identified based on GC-MS analysis can distinguish RCC patients from patients with other urinary cancers and healthy subjects, suggesting their potential as diagnostic markers for RCC.
8.Urinary metabolomics study of renal cell carcinoma based on gas chromatography-mass spectrometry
Lin ZHANG ; Ling LI ; Hairui KONG ; Fangyin ZENG
Journal of Southern Medical University 2015;(5):763-766
Objective To identify the biomarkers of renal cell cancer (RCC) through urine metabolic analysis. Methods Urine samples of 27 RCC patients, 26 patients with other urinary cancers and 26 healthy volunteers were examined with gas chromatography-mass spectrometry (GC-MS). SIMCA-P+12.0.1.0 software was used for principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA) to screen for the differential metabolites. Results PCA (R2X=0.846, Q2=0.575) and OPLS-DA (R2X=0.736, R2Y=0.974, Q2Y=0.897) model were established for the RCC patients and control subjects. Fourteen metabolites were selected as the characteristic metabolites, including pentanoic acid, malonic acid, glutaric acid, adipic acid, amino quinoline, quinoline, indole acetic acid, and tryptophan, whose levels in the urine were significantly higher in the RCC patients than in the normal subjects (P<0.01); the RCC patients showed significantly higher urine contents of pentanoic acid, phenylalanine, and 6-methoxy-nitro quinoline than those with other urinary tumors (P<0.01). Conclusion The urine metabolites identified based on GC-MS analysis can distinguish RCC patients from patients with other urinary cancers and healthy subjects, suggesting their potential as diagnostic markers for RCC.
9.Current status of physical activities among preschool children and relationship of activity type with bone mass
Ju ZHANG ; Fangyin WU ; Guo ZENG ; Li ZHANG ; Qiaoying ZENG
Chinese Journal of Health Management 2013;(3):159-162
Objective To investigate physical activities of preschool children by gender and to explore the effects of activity type on bone indexes.Methods During 2009 and 2010,397 preschool children of 3-5 years old were randomly selected from 4 kindergartens in Chengdu Province of China.Ultrasound bone analyzer was used to assess children's bone mass.A physical activity questionnaire was completed by parents to evaluate physical activities at leisure time.Student's t test and least square regression were used for data analysis.Results Concerning activity types,boys spent more leisure time on running and Wushu than girls did (t values were 1.94 and 2.84,respectively ; both P < 0.05).However,girls spent more time on dancing (0.78 h),jumping rope (0.08 h) and manual labour (0.22 h) each day (t values were-9.50,-3.43 and-1.92,respectively; all P < 0.05).The weekly total exercise time and energy consumption per unit of body weight of girls vs.boys were 7.29 vs.6.51 h and 127.57 vs.113.85 kJ (t values were 2.63 and 2.04,respectively ; both P < 0.05).About per day time on sleeping and per week time on watching television,there were no significant difference between boys and girls (t =0.180,0.520;P >0.05).But boys spent more time on electronic game and computer than girls (t =0.760,2.510;P < 0.05).The normalized correlation coefficient for bone mass and moderate physical activities or jumping was 0.184 and 0.275,respectively (both P < 0.05).Conclusions Our data suggest that preschool children's bone volume may be positively correlated with moderate physical activities and jumping activities.Introducing some moderate physical activities or activity appliance,toys and playing fields as well as increasing professional training might be helpful.
10.Partial cystectomy in treatment of localized muscle invasive bladder cancer
Yang ZHAO ; Guiping CHEN ; Hua WANG ; Fangyin LI ; Zongping WANG ; Shaoxing ZHU
Chinese Journal of Urology 2013;(7):497-500
Objective To evaluate the efficacy of partial cystectomy in treatment of localized muscle invasive bladder cancer.Methods From 1999 to 2005,data from 71 patients with muscle invasive bladder cancer(MIBC) were reviewed.There were 47 patients underwent partial cystectomy (PC) and 24 underwent total cystectomy (TC).The overall survival and disease-free survival in patients with MIBC with PC or TC were compared.All patients had pathologic T2-T3.Matched Kaplan-Meier survival analyses compared the effect of PC vs.TC on overall survival and disease-free survival.Univariate (log rank) and multivariate (Cox' proportional hazard model) analyses were used to test the statistical significance of several potential prognostic factors for survival rate.Results In the entire cohort,the overall survival rate and disease-free survival rate estimated at 5 years were 57% and 50% for PC patients,53% and 46% for TC patients,respectively (P>0.05).On univariate analysis,T stage (include vessel tumor embolus) and whether the tumor was pedunculated were the significant predictors of tumor recurrence.Age,gender,tumor quantity,tumor size and histology category were not associated with prognosis.Cox proportional hazard regression model confirmed that the independent prognosis factors of tumor was T stage (EXP(B)=1.64,P<0.05).Conclusions PC might not undermine cancer control in appropriately selected patients with MIBC.

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