Preparation of the Water-Soluble Fluorene-Containing Fluorescent Polymer by One-Pot Method

Yuan Wang,Hongchi Zhao,Haijian Tan,Xiaomeng Li,Yunxia Qian,Libin Bai,Yonggang Wu,Shufang Lv 한국고분자학회 2015 Macromolecular Research Vol.23 No.10

A water-soluble fluorescent polymer, 2,7-di{4'-[3''-(S-poly(acryloyl ethylene diamine hydrochloride)-2'''- methyl propionic acid)propionyloxy hexyloxy]phenyl}-9,9-di(n-octyl)fluorene (P3) with regular and ordered structure, was designed and synthesized with high conversion using click reaction between thiols and carbon-carbon double bonds. Because the S-H bond is relatively weak, thiol groups are subject to rapid oxidation during purification and storage and thereby result in the low conversion in the following click reaction. To circumvent this limitation, the synthesis of thiol groups and click reaction were simultaneously performed in one system (one-pot method). The intermediate products P1 (S-1-dodecyl-S'-[poly(N-Boc-acryloyl ethylene diamine)-2'-methyl propionic acid]trithiocarbonate) and M2 (2,7-di[4-(acryloyloxy hexyloxy)phenyl]-9,9-di(n-octyl)fluorene) were prepared via reversible addition fragmentation chain transfer (RAFT) polymerization and Williamson synthesis, respectively. Then, 2,7- di{4'-[3''-(S-poly(N-Boc-acryloyl ethylene diamine)-2'''-methyl propionic acid)propionyloxy hexyloxy]phenyl}- 9,9-di(n-octyl)fluorene (P2) was synthesized with high conversion (83%) by employing a click reaction with P1 and M2 as starting materials. Finally, water-soluble fluorescent polymer P3 was obtained after the deprotection of P2. M2, P1, P2 and P3 were characterized by Fourier transform infrared (FTIR) spectrum, nuclear magnetic resonance (NMR), high-resolution mass spectroscopy (MS), gel permeation chromatograph (GPC), UV-Visible (UV-Vis) spectrum, and fluorescence, respectively. The results revealed that P3 displayed good solubility in water and not only exhibited a blue fluorescence emission band in water but also had a similar photoluminescent spectra to those of M2 and P2 in dichloromethane. The fluorescence quantum yield of P3 in aqueous solution could reach 0.10. In addition, the result of cell cytotoxicity indicates that P3 has low cytotoxicity and exhibits potential for biomedical applications. This work provides a new insight into the study of water-soluble fluorescent polymer.

Current Status of Serum Insulin and C-Peptide Measurement in Clinical Laboratories: Experience from 94 Laboratories in China

Zhou Weiyan,Deng Yuhang,Zhao Haijian,Zhang Chuanbao 대한진단검사의학회 2022 Annals of Laboratory Medicine Vol.42 No.4

Background: Accurate measurements of serum insulin and C-peptide are needed for the therapy and classification of diabetes. This study investigated the status of serum insulin and C-peptide measurements in China by analyzing the results of five pooled serum samples measured in 94 laboratories. Methods: Patient serum samples were pooled into five groups according to insulin and C-peptide concentrations and measured in 94 laboratories using different measurement systems. The inter- and intra-laboratory %CV as well as inter- and intra-measurement system %CV were calculated to assess the status of insulin and C-peptide measurements. To verify whether the disagreement between laboratories was due to different calibrators, as reported in previous studies, one low-level and one high-level sample extracted from the five pooled serum samples were used to recalibrate clinical measurement systems. Results: The mean intra-laboratory, intra-measurement system, inter-laboratory, and inter-measurement system %CVs were 2.7%, 4.8%, 21.8%, and 22.4%, respectively, for insulin and 2.3%, 6.7%, 16.4%, and 24.5%, respectively, for C-peptide. The inter- and intra-laboratory %CVs for insulin decreased with increasing concentration. After recalibration with low- and high-level samples, the mean inter-measurement %CV decreased from 22.4% to 17.2% for insulin and from 24.5% to 5.7% for C-peptide. Conclusions: The intra-laboratory and intra-measurement system imprecision values are satisfactory for serum insulin and C-peptide measurements. However, the results from laboratories using different measurement systems were not comparable, and there is still much work needed to achieve the standardization or harmonization of serum insulin and C-peptide measurements.

MULTIVARIATE EFFECTIVENESS OF ECOLANE AND ECOHMI BASED COOPERATIVE VEHICLE-INFRASTRUCTURE SYSTEM

Qiang Fu,Yiping Wu,Xiaohua Zhao,Yang Bian,Li HaijianLi 한국자동차공학회 2023 International journal of automotive technology Vol.24 No.1

The cooperative vehicle-infrastructure system (CVIS) offers opportunities to further enhance the potential of energy saving and emission reduction for eco-driving. However, it is still not clear about the effectiveness of CVIS on promoting eco-driving nor the multivariate effectiveness of CVIS designed for eco-driving on traffic flow and safety characteristics. A dedicated ecological lane (Ecolane) and ecological human-machine-interaction (EcoHMI) based CVIS (Ecolane-HMI-CVIS) were developed based on driving simulator technology. An experiment of 35 participants was conducted to study eco-driving behavior and final utility to identify the influence of Ecolane-HMI-CVIS. The results indicated that the Ecolane-HMI-CVIS enhanced eco-driving behavior and reduced emission, with CO and NOX significantly reduced by 10.72 % and 9.83 % respectively. The Ecolane-HMI-CVIS reduced the headway and promoted vehicle operation stability, ordering, and improved traffic capacity about 10 %. No negative impact of Ecolane-HMI-CVIS was observed on traffic safety. This study developed a test platform based on driving simulator to explore the multivariate influence of Ecolane-HMI-CVIS. In addition to analyzing the overall effect of Ecolane-HMI-CVIS during the whole experimental section, its detailed influence at each key zone and in spatial change process were also analyzed. This research contributed to better understanding of the working mechanism and effectiveness of Ecolane-HMI-CVIS, and provide technical and policy references of CVIS based eco-driving for traffic management departments.

Evaluation of Coefficients of Variation for Clinical Chemistry Tests Based on Internal Quality Control Data Across 5,425 Laboratories in China From 2013 to 2022

Wang Wei,Zhang Zhixin,Zhang Chuanbao,Zhao Haijian,Yuan Shuai,Liu Jiali,Dong Na,Wang Zhiguo,Kang Fengfeng 대한진단검사의학회 2024 Annals of Laboratory Medicine Vol.44 No.3

Background: Clinical chemistry tests are most widely used in clinical laboratories, and diverse measurement systems for these analyses are available in China. We evaluated the imprecision of clinical chemistry measurement systems based on internal QC (IQC) data. Methods: IQC data for 27 general chemistry analytes were collected in February each year from 2013 to 2022. Four performance specifications were used to calculate pass rates for CVs of IQC data in 2022. Boxplots were drawn to analyze trends of CVs, and differences in CVs among different groups were assessed using the Mann–Whitney U-test or Kruskal–Wallis test. Results: The number of participating laboratories increased significantly from 1,777 in 2013 to 5,425 in 2022. CVs significantly decreased for all 27 analytes, except creatine kinase and lipase. Triglycerides, total bilirubin, direct bilirubin, iron, and γ-glutamyl transferase achieved pass rates >80% for all goals. Nine analytes with pass rates <80% based on 1/3 allowable total error were further analyzed; the results indicated that closed systems exhibited lower CVs than open systems for all analytes, except total protein. For all nine analytes, differences were significant between tertiary hospitals and non-tertiary hospitals and between accredited and non-accredited laboratories. Conclusions: The CVs of IQC data for clinical chemistry have seen a continuous overall improvement in China. However, there is ample room for imprecision improvement for several analytes, with stricter performance specifications.

Commutability Assessment of Processed Human Plasma Samples for Normetanephrine and Metanephrine Measurements Based on the Candidate Reference Measurement Procedure

Deng Yuhang,Liu Qingxiang,Liu Zhenni,Zhao Haijian,Zhou Weiyan,Zhang Chuanbao 대한진단검사의학회 2022 Annals of Laboratory Medicine Vol.42 No.5

Background: To identify candidate external quality assessment (EQA) materials for normetanephrine and metanephrine measurements, we assessed the commutability of eight processed human plasma samples. The agreement between routine assays and the candidate reference measurement procedure (cRMP) was also evaluated. Methods: Fifty-three clinical samples and eight processed plasma samples were prepared. The processed samples included pooled and individual plasma samples spiked with pure normetanephrine and metanephrine and non-spiked pooled and individual plasma samples. The clinical and processed samples were subjected to four routine isotope dilution tandem mass spectrometry assays and cRMP. Commutability was assessed based on two approaches recommended by the CLSI and International Federation of Clinical Chemistry (IFCC). Passing–Bablok regression and Bland–Altman analysis were used to evaluate the agreement between the routine assays and cRMP. Results: The commutability results of the CLSI approach were better than those of the IFCC approach. For the CLSI approach, spiked individual plasma samples and spiked high-concentration pooled plasma samples were commutable for all routine assays for both analytes. The non-spiked pooled plasma sample was commutable for two out of four routine assays for metanephrine and three out of four routine assays for normetanephrine. The agreement between the routine assays and the cRMP was satisfactory, except for one routine assay showing significant bias. Conclusions: High-concentration spiked pooled plasma samples and spiked individual plasma samples are candidate EQA materials for normetanephrine and metanephrine measurements.

A putative pathogen-resistant regulatory pathway between MicroRNAs and candidate target genes in maize

Jian Gao,Mao Luo,Chun Zhang,Hua Peng,Haijian Lin,Yaou shen,Maojun Zhao,Guangtang Pan,Zhiming Zhang 한국식물학회 2015 Journal of Plant Biology Vol.58 No.4

MicroRNAs (miRNAs) are a family of small non-coding RNAs that, in most cases, negatively regulate gene expression at the post-transcriptional level. Plant miRNAs have been implicated in developmental processes and adaptation to environmental stress including biotic and abiotic stresses. Here, we report a comprehensive analysis of miRNAs and associated target genes under banded leaf and sheath blight (BLSB) stress caused by R. solani in maize. Eight differentially expressed miRNAs were randomly selected from deep sequencing results and validated by qRT-PCR together with their putative target genes, most of which are transcription factors as well as metabolic genes involved in auxin signaling. The results revealed that majorities of the analyzed miRNAs show an inverse correlation with their corresponding predicted target genes. In addition, a putative regulatory network of miRNAs-mRNAs responsive to R. solani was constructed. This study provides insight into the regulatory functions of miRNAs, thereby expanding our knowledge of the molecular mechanisms of pathogen resistance.

An Accurate Isotope Dilution Liquid Chromatography-Tandem Mass Spectrometry Method for Serum C-Peptide and Its Use in Harmonization in China

Deng Yuhang,Zhang Chao,Wang Jing,Zeng Jie,Zhang Jiangtao,Zhang Tianjiao,Zhao Haijian,Zhou Weiyan,Zhang Chuanbao 대한진단검사의학회 2023 Annals of Laboratory Medicine Vol.43 No.4

Background: Serum C-peptide results from various routine methods used in China are highly variable, warranting well-performing methods to serve as an accuracy base to improve the harmonization of C-peptide measurements in China. We developed an accurate isotope dilution liquid chromatography-tandem mass spectrometry (ID-LC–MS/MS) method for serum C-peptide measurement and explored its use in harmonization. Methods: After protein precipitation with ZnSO4 solution, C-peptide was extracted from serum samples by anion-exchange solid-phase extraction and quantified by ID-LC–MS/MS in positive ion mode. The precision and analytical recovery of the ID-LC–MS/MS method were assessed. Seventy-six serum samples were analyzed using the ID-LC–MS/MS method and six routine immunoassays. Ordinary linear regression (OLR) and Bland-Altman (BA) analyses were conducted to evaluate the relationship between the ID-LC–MS/MS method and routine immunoassays. Five serum pool samples assigned using the ID-LC–MS/MS method were used to recalibrate the routine assays. OLR and BA analyses were re-conducted after recalibration. Results: The within-run, between-run, and total precision for the ID-LC–MS/MS method at four concentrations were 1.0%–2.1%, 0.6%–1.2%, and 1.3%–2.2%, respectively. The analytical recoveries for the ID-LC–MS/MS method at three concentrations were 100.3%–100.7%, 100.4%–101.0%, and 99.6%–100.7%. The developed method and the immunoassays were strongly correlated, with all R2 >0.98. The comparability among the immunoassays was substantially improved after recalibration. Conclusions: The performance of the ID-LC–MS/MS method was carefully validated, and this method can be used to improve the harmonization of serum C-peptide measurements in China.

Commutability Assessment of Candidate External Quality Assessment Materials for Aminotransferase Activity Measurements Based on Different Approaches in China

Long Qichen,Qi Tianqi,Zhang Tianjiao,Wang Jing,Zeng Jie,Ying Yan,Meng Wang,Wei Huang,Haijian Zhao,Wenxiang Chen,Chuanbao Zhang 대한진단검사의학회 2021 Annals of Laboratory Medicine Vol.41 No.1

Background: Using commutable external quality assessment (EQA) materials is important for monitoring successful harmonization efforts. We assessed the commutability of four human serum pool (HSP) preparations to identify candidate EQA materials for alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activity measurement. Methods: One set each of 85 clinical samples (CSs) was collected for ALT and AST activity measurement. The 15 candidate EQA materials included four types of HSP preparations (A to D): materials A, C, and D contained human original recombinant (HOR) aminotransferases; materials B was mixed leftover samples. The CSs and 15 candidate EQA materials were analyzed using seven routine assays, and the ln-transformed results were analyzed in 21 assay pairs. Commutability was assessed using Deming regression, with a 95% prediction interval (CLSI approach) and the difference in bias with an error component model (International Federation of Clinical Chemistry and Laboratory Medicine [IFCC] approach). Results: For ALT, all materials were commutable for 14–21 assay pairs according to the CLSI and IFCC approaches. For AST, B01-03 showed commutability for 14-21 assay pairs, and C01-03 and D01-03 showed commutability for no less than 10 assay pairs according to the two approaches. A01-06 were commutable for 9-16 assay pairs according to the CLSI approach, but for 6-9 assay pairs according to the IFCC approach. Conclusions: Mixed leftover samples showed desirable commutability characteristics as candidate EQA materials for routine aminotransferase activity measurements. Human serum bases supplemented with HOR were commutable for most routine ALT activity measurements.