1.Experimental Study on Expression of Carbapenem Resistant Pseudomonas Aeruginosa's 10 Membrane Protein Coding Genes in Kunming
Zan LU ; Hongyan ZHAO ; Chunfu LI ; Limin YIN ; Baojun REN ; Guibo SONG ; Xu YANG
Journal of Modern Laboratory Medicine 2025;40(1):7-12
Objective To understand the membrane protein molecular epidemiology of carbapenem-resistant Pseudomonas aeruginosa (CRPA) in the region,and provide some evidence for rational drug use or application of efflux pump inhibitors. Methods Collected Pseudomonas aeruginosa isolated from four hospitals in the region from October 2022 to August 2023,and used SYBR-PCR method to quantitatively detect the relative mRNA expression (RE) levels of 10 membrane protein coding genes,including mexA,B,C,D,E,F,X,Y,and oprD,M. Then categorized the strains into five groups based on ceftazidime,cefepime,imipenem,and meropenem resistance phenotype combination,including the compassionate group (Group Ⅰ),Group Ⅱ with full resistance,IPM,MEM resistant,CAZ and CFP sensitive groups (Group Ⅲ),IPM resistance,MEM non-resistance (sensitive or intermediate) group (Group Ⅳ),IPM,MEM resistance,CAZ and CFP non-resistance groups (Group V).The median RE of each membrane protein-coding gene was analyzed. Results A total of 108 strains of Pseudomonas aeruginosa were collected,with 24 strains in Group Ⅰ as controls and 84 strains in the carbapenem resistant group,including 32 strains in Group Ⅱ,22 strains in Group Ⅲ,13 strains in Group Ⅳ,and 17 strains in Group Ⅴ. The expression of mexD,mexE,mexF,mexX and mexY in the drug-resistant group was higher than that in the control group,and the differences were statistically significant (U=409.5~661.0,all P<0.05). There was no statistically significant difference in mexA,mexB,mexC,oprD and oprM with the control group (U=767.0~1004.5,all P>0.05). There was no significant difference in the expression of RE genes encoding various membrane proteins among strains from different hospitals (H=0.914~7.407,all P>0.05). Among the four different phenotypes,there was no statistically significant difference in the irregular distribution of mexA and oprM RE between each group and the control group (UmexA=95.0~264.0,UoprM=143.0~331.0). The mexC RE in each group was lower than that in the control group,but the differences were not statistically significant (U=134.0~344.5,all P>0.05). MeixE and meixY RE were both higher than the control group,and the differences were statistically significant (UmexE=48.0~230.0,UmexY=83.0~184.0). MeixB was lower than the control group in group Ⅳ (U=72.0),and the differences were statistically significant (all P<0.05). MeixD and meixF showed consistent expression,with higher expression in groups Ⅲ,Ⅳ and Ⅴ compared to the control group (UmeixD=34.0~102.0,UmeixF=65.0~113.0). MeixX was expressed higher in groups Ⅱ,Ⅳ and Ⅴ compared to the control group (U=164.0,58.0,111.0),while oprD was only expressed lower in group Ⅲ than in the control group (U=140.0),with statistically significant differences (all P<0.05). Although the expression of oprD in groups Ⅱ,Ⅳ and Ⅴ was lower than that in the control group,the differences were not statistically significant (U=381.0,102.0,144.0,all P>0.05). Conclusion ExCD,mexEF and mexXY are the main membrane protein combinations of CRPA efflux pumps in Kunming area. Upregulation of mexD,E,F,X,and Y membrane protein expression enhanced efflux. The correlation between mexAB oprM efflux pump and carbapenem resistance in CRPA in this area was low. The low expression of oprD played a role in the efflux mechanism in strains that do not produce β-lactase,but there was no significant difference in low expression in enzyme producing strains.
2.Experimental Study on Expression of Carbapenem Resistant Pseudomonas Aeruginosa's 10 Membrane Protein Coding Genes in Kunming
Zan LU ; Hongyan ZHAO ; Chunfu LI ; Limin YIN ; Baojun REN ; Guibo SONG ; Xu YANG
Journal of Modern Laboratory Medicine 2025;40(1):7-12
Objective To understand the membrane protein molecular epidemiology of carbapenem-resistant Pseudomonas aeruginosa (CRPA) in the region,and provide some evidence for rational drug use or application of efflux pump inhibitors. Methods Collected Pseudomonas aeruginosa isolated from four hospitals in the region from October 2022 to August 2023,and used SYBR-PCR method to quantitatively detect the relative mRNA expression (RE) levels of 10 membrane protein coding genes,including mexA,B,C,D,E,F,X,Y,and oprD,M. Then categorized the strains into five groups based on ceftazidime,cefepime,imipenem,and meropenem resistance phenotype combination,including the compassionate group (Group Ⅰ),Group Ⅱ with full resistance,IPM,MEM resistant,CAZ and CFP sensitive groups (Group Ⅲ),IPM resistance,MEM non-resistance (sensitive or intermediate) group (Group Ⅳ),IPM,MEM resistance,CAZ and CFP non-resistance groups (Group V).The median RE of each membrane protein-coding gene was analyzed. Results A total of 108 strains of Pseudomonas aeruginosa were collected,with 24 strains in Group Ⅰ as controls and 84 strains in the carbapenem resistant group,including 32 strains in Group Ⅱ,22 strains in Group Ⅲ,13 strains in Group Ⅳ,and 17 strains in Group Ⅴ. The expression of mexD,mexE,mexF,mexX and mexY in the drug-resistant group was higher than that in the control group,and the differences were statistically significant (U=409.5~661.0,all P<0.05). There was no statistically significant difference in mexA,mexB,mexC,oprD and oprM with the control group (U=767.0~1004.5,all P>0.05). There was no significant difference in the expression of RE genes encoding various membrane proteins among strains from different hospitals (H=0.914~7.407,all P>0.05). Among the four different phenotypes,there was no statistically significant difference in the irregular distribution of mexA and oprM RE between each group and the control group (UmexA=95.0~264.0,UoprM=143.0~331.0). The mexC RE in each group was lower than that in the control group,but the differences were not statistically significant (U=134.0~344.5,all P>0.05). MeixE and meixY RE were both higher than the control group,and the differences were statistically significant (UmexE=48.0~230.0,UmexY=83.0~184.0). MeixB was lower than the control group in group Ⅳ (U=72.0),and the differences were statistically significant (all P<0.05). MeixD and meixF showed consistent expression,with higher expression in groups Ⅲ,Ⅳ and Ⅴ compared to the control group (UmeixD=34.0~102.0,UmeixF=65.0~113.0). MeixX was expressed higher in groups Ⅱ,Ⅳ and Ⅴ compared to the control group (U=164.0,58.0,111.0),while oprD was only expressed lower in group Ⅲ than in the control group (U=140.0),with statistically significant differences (all P<0.05). Although the expression of oprD in groups Ⅱ,Ⅳ and Ⅴ was lower than that in the control group,the differences were not statistically significant (U=381.0,102.0,144.0,all P>0.05). Conclusion ExCD,mexEF and mexXY are the main membrane protein combinations of CRPA efflux pumps in Kunming area. Upregulation of mexD,E,F,X,and Y membrane protein expression enhanced efflux. The correlation between mexAB oprM efflux pump and carbapenem resistance in CRPA in this area was low. The low expression of oprD played a role in the efflux mechanism in strains that do not produce β-lactase,but there was no significant difference in low expression in enzyme producing strains.
3.Temporal-spatial Generation of Astrocytes in the Developing Diencephalon.
Wentong HONG ; Pifang GONG ; Xinjie PAN ; Zhonggan REN ; Yitong LIU ; Guibo QI ; Jun-Liszt LI ; Wenzhi SUN ; Woo-Ping GE ; Chun-Li ZHANG ; Shumin DUAN ; Song QIN
Neuroscience Bulletin 2024;40(1):1-16
Astrocytes are the largest glial population in the mammalian brain. However, we have a minimal understanding of astrocyte development, especially fate specification in different regions of the brain. Through lineage tracing of the progenitors of the third ventricle (3V) wall via in-utero electroporation in the embryonic mouse brain, we show the fate specification and migration pattern of astrocytes derived from radial glia along the 3V wall. Unexpectedly, radial glia located in different regions along the 3V wall of the diencephalon produce distinct cell types: radial glia in the upper region produce astrocytes and those in the lower region produce neurons in the diencephalon. With genetic fate mapping analysis, we reveal that the first population of astrocytes appears along the zona incerta in the diencephalon. Astrogenesis occurs at an early time point in the dorsal region relative to that in the ventral region of the developing diencephalon. With transcriptomic analysis of the region-specific 3V wall and lateral ventricle (LV) wall, we identified cohorts of differentially-expressed genes in the dorsal 3V wall compared to the ventral 3V wall and LV wall that may regulate astrogenesis in the dorsal diencephalon. Together, these results demonstrate that the generation of astrocytes shows a spatiotemporal pattern in the developing mouse diencephalon.
Mice
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Animals
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Astrocytes
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Neuroglia/physiology*
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Diencephalon
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Brain
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Neurons
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Mammals
4.The applications and prospects of non-coding RNAs in the diagnosis and treatment of heart failure
Chinese Journal of Laboratory Medicine 2019;42(8):613-617
Heart failure (HF) is a severe or advanced stage of heart disease with high mortality and readmission rates. The detection of biomarkers plays an important role in the diagnosis, treatment and clinical management of HF. In addition to the traditional biomarkers, the non-coding RNA markers such as miRNA, circRNA, and lncRNA have also shown better future applications in the diagnosis and differential diagnosis of HF, the severity and prognosis assessment, the risk assessment of cardiovascular events after discharge in patients with HF. These biomarkers will also contribute to personalized clinical management of HF.
5.The monitoring and analysis results of 15 clinical laboratory quality indicators from 2011 to 2015
Chong GUO ; Zijie LIU ; Guibo SONG ; Xin LI ; Xiao WANG ; Yong DUAN
Chinese Journal of Laboratory Medicine 2016;(1):29-33
Objective To investigate how to improve test quality by monitoring and analyzing 15 clinical laboratory quality indicators from the National Health and Family Planning Commission .Methods Data were collected from clinical laboratory department of the First Affiliated Hospital of Kunming Medical University between January 2011 and August 2015.15 quality indicators were analyzed retrospectively , including the error rate of specimen type , the coefficient variation unqualified rate of internal quality control test, the reporting rate of critical value , et al.Results The monitoring results of quality indicators basically satisfied the quality goals , except that the median of turn around time in pre-analytical phase was not established, routine internal quality control was not conducted in some laboratory tests in analytical phase and the reporting rate and reporting timely rate of critical value should be further improved in post -analytical phase .Conclusion Medical laboratory quality system can be continuously improved by means of setting up the quality goals of 15 quality indicators referring to sub-specialty and laboratory tests , as well as automated monitoring, statistics and analysis in LIS.
6.Analysis of genomic copy number variations in two sisters with primary amenorrhea and hyperandrogenism.
Yanliang ZHANG ; Qiuyue XU ; Xuemei CAI ; Yixun LI ; Guibo SONG ; Juan WANG ; Rongchen ZHANG ; Yong DAI ; Yong DUAN
Chinese Journal of Medical Genetics 2015;32(6):814-818
OBJECTIVETo analyze genomic copy number variations (CNVs) in two sisters with primary amenorrhea and hyperandrogenism.
METHODSG-banding was performed for karyotype analysis. The whole genome of the two sisters were scanned and analyzed by array-based comparative genomic hybridization (array-CGH). The results were confirmed with real-time quantitative PCR (RT-qPCR).
RESULTSNo abnormality was found by conventional G-banded chromosome analysis. Array-CGH has identified 11 identical CNVs from the sisters which, however, overlapped with CNVs reported by the Database of Genomic Variants (http://projects.tcag.ca/variation/). Therefore, they are likely to be benign. In addition, a -8.44 Mb 9p11.1-p13.1 duplication (38,561,587-47,002,387 bp, hg18) and a -80.9 kb 4q13.2 deletion (70,183,990-70,264,889 bp, hg18) were also detected in the elder and younger sister, respectively. The relationship between such CNVs and primary amenorrhea and hyperandrogenism was however uncertain. RT-qPCR results were in accordance with array-CGH.
CONCLUSIONTwo CNVs were detected in two sisters by array-CGH, for which further studies are needed to clarify their correlation with primary amenorrhea and hyperandrogenism.
Amenorrhea ; diagnosis ; genetics ; Chromosomes, Human, Pair 4 ; genetics ; Chromosomes, Human, Pair 9 ; genetics ; Comparative Genomic Hybridization ; methods ; DNA Copy Number Variations ; genetics ; Female ; Humans ; Hyperandrogenism ; diagnosis ; genetics ; Karyotyping ; Reverse Transcriptase Polymerase Chain Reaction ; Siblings ; Young Adult

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