Large Instrument- and Detergent-Free Assay for Ultrasensitive Nucleic Acids Isolation via Binary Nanomaterial

Liu, Huifang,Zhao, Fei,Jin, Choong Eun,Koo, Bonhan,Lee, Eun Yeong,Zhong, Linlin,Yun, Kyusik,Shin, Yong American Chemical Society 2018 ANALYTICAL CHEMISTRY - Vol.90 No.8

<P>Nucleic acid-based diagnostics are widely used for clinical applications due to their powerful recognition of biomolecule properties. Isolation and purification of nucleic acids such as DNA and RNA in the diagnostic system have been severely hampered in point-of-care testing because of low recovery yields, degradation of nucleic acids due to the use of chaotropic detergent and high temperature, and the requirement of large instruments such as centrifuges and thermal controllers. Here, we report a novel large instrument- and detergent-free assay via binary nanomaterial for ultrasensitive nucleic acid isolation and detection from cells (eukaryotic and prokaryotic). This binary nanomaterial couples a zinc oxide nanomultigonal shuttle (ZnO NMS) for cell membrane rupture without detergent and temperature control and diatomaceous earth with dimethyl suberimidate complex (DDS) for the capture and isolation of nucleic acids (NA) from cells. The ZnO NMS was synthesized to a size of 500 nm to permit efficient cell lysis at room temperature within 2 min using the biological, chemical, and physical properties of the nanomaterial. By combining the ZnO NMS with the DDS and proteinase K, the nucleic acid extraction could be completed in 15 min with high quantity and quality. For bacterial cells, DNA isolation with the binary nanomaterial yielded 100 times more DNA, than a commercial spin column based reference kit, as determined by the NanoDrop spectrophotometer. We believe that this binary nanomaterial will be a useful tool for rapid and sensitive nucleic acid isolation and detection without large instruments and detergent in the field of molecular diagnostics.</P> [FIG OMISSION]</BR>

Pharmacokinetics and Biodistribution of Surface Modification Polymeric Nanoparticles

Liu, Mingxing,Li, Huifang,Luo, Guoan,Liu, Qingfei,Wang, Yiming 대한약학회 2008 Archives of Pharmacal Research Vol.31 No.4

The objective of this study is to investigate the pharmacokinetics and biodistribution of free breviscapine (BVP) and coated BVP-loaded poly (D, L-lactic acid) nanoparticles (BVP-PLA-NPs) in rats after i.v. administration. Coated BVP-PLA-NPs were prepared by the spontaneous emulsification solvent diffusion method and characterized. The BVP content in the NPs, the biological samples and in vitro release was measured by the high-performance liquid chromatography (HPLC). The mean sizes of coated BVP-PLA-NPs were 177 and 319 nm with a narrow distribution and smooth sphere shapes, entrapment efficiency of 86.9% and 93.1%, respectively. Drug release profiles in phosphate buffer and plasma exhibited a biphasic release phenomenon. After i.v. administration of free BVP and NPs suspensions in rats, area under plasma concentration-time curve and elimination $t_{1/2}$ were increased 9.3-fold and 10.9-fold for 177 nm of NPs, and 4.4-fold and 17.1-fold for 319 nm of NPs compared with that of free BVP, respectively. NPs were mainly distributed in liver, spleen, heart and brain. In addition, NPs could penetrate blood brain barrier (BBB) and the particle size had some effect on pharmacokinetics and biodistribution. Coated BVP-PLA-NPs could effectively avoid the capture by the reticuloendothelial system and prolong the half-life of BVP. Moreover, these NPs could penetrate BBB and enhance the accumulation of BVP in brain.

Synthesis, Characterization, and Antibacterial Properties of Silver Nanoparticles-Graphene and Graphene Oxide Composites

Huifang Liu,Linlin Zhong,윤규식,Monica Samal 한국생물공학회 2016 Biotechnology and Bioprocess Engineering Vol.21 No.1

In the field of nanotechnology, silver nanoparticles have been considered a promising antibacterial material for a century. The potential applications of graphene-based materials are increasingly recognized for their special physico-chemical and biological properties. In particular, graphene and graphene oxide as the foundation of nanocomposites have garnered much interest among researchers in many fields. In this review, we concentrate on different aspects of silver nanoparticle composites with graphene and graphene oxide, focusing on their synthesis methods, special characteristics, and antibacterial properties; we also briefly discuss limitations and future research.

Pharmacokinetics and Biodistribution of Surface Modification Polymeric Nanoparticles

Mingxing Liu,Huifang Li,Guoan Luo,Qingfei Liu,Yiming Wang 대한약학회 2008 Archives of Pharmacal Research Vol.31 No.4

The objective of this study is to investigate the pharmacokinetics and biodistribution of free breviscapine (BVP) and coated BVP-loaded poly (D, L-lactic acid) nanoparticles (BVP-PLANPs) in rats after i.v. administration. Coated BVP-PLA-NPs were prepared by the spontaneous emulsification solvent diffusion method and characterized. The BVP content in the NPs, the biological samples and in vitro release was measured by the high-performance liquid chromatography (HPLC). The mean sizes of coated BVP-PLA-NPs were 177 and 319 nm with a narrow distribution and smooth sphere shapes, entrapment efficiency of 86.9% and 93.1%, respectively. Drug release profiles in phosphate buffer and plasma exhibited a biphasic release phenomenon. After i.v. administration of free BVP and NPs suspensions in rats, area under plasma concentration-time curve and elimination t1/2 were increased 9.3-fold and 10.9-fold for 177 nm of NPs, and 4.4-fold and 17.1-fold for 319 nm of NPs compared with that of free BVP, respectively. NPs were mainly distributed in liver, spleen, heart and brain. In addition, NPs could penetrate blood brain barrier (BBB) and the particle size had some effect on pharmacokinetics and biodistribution. Coated BVP-PLA-NPs could effectively avoid the capture by the reticuloendothelial system and prolong the half-life of BVP. Moreover, these NPs could penetrate BBB and enhance the accumulation of BVP in brain.

Association between Initial Chest CT or Clinical Features and Clinical Course in Patients with Coronavirus Disease 2019 Pneumonia

Liu Zhe,Jin Chao,Wu Carol C.,Liang Ting,Zhao Huifang,Wang Yan,Wang Zekun,Li Fen,Zhou Jie,Cai Shubo,Zeng Lingxia,Yang Jian 대한영상의학회 2020 Korean Journal of Radiology Vol.21 No.6

Objective: To identify the initial chest computed tomography (CT) findings and clinical characteristics associated with the course of coronavirus disease 2019 (COVID-19) pneumonia. Materials and Methods: Baseline CT scans and clinical and laboratory data of 72 patients admitted with COVID-19 pneumonia (39 men, 46.2 ± 15.9 years) were retrospectively analyzed. Baseline CT findings including lobar distribution, presence of ground glass opacities, consolidation, linear opacities, and lung severity score were evaluated. The outcome event was recovery with hospital discharge. The time from symptom onset to discharge or the end of follow-up (for those remained hospitalized) was recorded. Data were censored in events such as death or discharge without recovery. Multivariable Cox proportional hazard regression was used to explore the association between initial CT, clinical or laboratory findings, and discharge with recovery, whereby hazard ratio (HR) values < 1 indicated a lower rate of discharge at four weeks and longer time until discharge. Results: Thirty-two patients recovered and were discharged during the study period with a median length of admission of 16 days (range, 9 to 25 days), while the rest remained hospitalized at the end of this study (median, 17.5 days; range, 4 to 27 days). None died during the study period. After controlling for age, onset time, lesion characteristics, number of lung lobes affected, and bilateral involvement, the lung severity score on baseline CT (> 4 vs. ≤ 4 [reference]: adjusted HR = 0.41 [95% confidence interval, CI = 0.18–0.92], p = 0.031) and initial lymphocyte count (reduced vs. normal or elevated [reference]: adjusted HR = 0.14 [95% CI = 0.03–0.60], p = 0.008) were two significant independent factors that influenced recovery and discharge. Conclusion: Lung severity score > 4 and reduced lymphocyte count at initial evaluation were independently associated with a significantly lower rate of recovery and discharge and extended hospitalization in patients admitted for COVID-19 pneumonia.

Keratin-based drug-protein conjugate with acid-labile and reduction-cleavable linkages in series for tumor intracellular DOX delivery

Huifang Zhang,Mingliang Pei,Peng Liu 한국공업화학회 2019 Journal of Industrial and Engineering Chemistry Vol.80 No.-

PK-SS-Hy-D drug-protein prodrug was obtained by conjugating doxorubicin (DOX) onto the PEGylatedkeratin (PK) with both bioreducible disulfide linkage and acid-cleavable hydrazone bond in a seriesconnection mode. The controlled release profiles demonstrated the pH and reduction dual-responsivetriggered release of DOX from the proposed drug-protein conjugate nanoparticles, with a low prematuredrug leakage of 5.5% in the simulated physiological medium. The in vitro experiments indicated that theproposed prodrug nanoparticles could delivery DOX into the cell nuclei, with an enhanced anti-tumorefficacy. It is expected as a potential candidate for future tumor chemotherapy with minimized toxic sideeffect.

The Core Knowledge and Skills of Nursing Competency Regarding Mealtime Assistance for Hemiplegic Patients in China

Chen Huifang,Nakatani Hisae,Liu Ting,Zhao Haiwei,Xie Di 한국간호과학회 2020 Asian Nursing Research Vol.14 No.2

Purpose: Hemiplegic patients often experience malnutrition and feeding risks due to disabilities and inadequate nursing support; nursing roles regarding mealtime assistance remain unclear in China. Therefore, this study aimed to clarify the core knowledge and skills of nursing competency regarding mealtime assistance for hemiplegic patients in China. Methods: A cross-sectional survey of self-administered questionnaire to 640 nurses (response rate: 57.7%) from two tertiary teaching hospitals and two sanatoriums was conducted. Survey content included 25 items regarding the mealtime assistance competency plus 6 items on demographic characteristics. The factor structure of the 25 items was explored and verified by exploratory and confirmatory factor analysis. Its reliability was confirmed by Cronbach's alpha. Results: Three factors including 22 items with a cumulative rate of 60.9% were identified: “Assistance knowledge and skills for acute period”, “Knowledge about assistance and guidance for recovery period,” and “Professional basic knowledge regarding hemiplegia.” Its reliability was ensured with Cronbach's alpha ranging from .86 to .96. In addition, evidence for its construct validity was obtained, as structural equation modeling revealed a good fit to the data within the allowable range based on various fit indices. Conclusion: This study clarified the core knowledge and skills regarding mealtime assistance competency for hemiplegic patients, with “Assistance for acute period” most concerned, followed by “assistance and guidance for recovery period,” and “Professional basic knowledge,” and the latter two need more attention. Obtained results can provide useful evidence for competent nursing practice to improve the quality of mealtime assistance for hemiplegic patients in China.

Temporal distribution, influencing factors and pollution sources of urban ambient air quality in Nanchong, China

Hong Zhou,Youping Li,Huifang Liu,Zhongyu Fan,Jie Xia,Shanli Chen,Yuxiang Zheng,Xiaocui Chen 대한환경공학회 2015 Environmental Engineering Research Vol.20 No.3

The PM10, SO₂ and NO₂ mass concentrations were obtained over five years from monitoring stations across Nanchong, a southwest city in China. Changes in urban air quality over time, as well as the factors influencing that change, were evaluated based on air pollutant concentrations, the Air Pollution Index (API), and the Comprehensive Pollution Index (P). The results showed that the total annual mean PM10, SO₂ and NO₂ concentrations over the five years studied were 61.1±1.1, 45.0±3.9 and 34.9±4.9 μg·m-3, respectively. The annual mean concentrations displayed a generally decreasing trend; lower than the annual mean second-level air quality limit. Meanwhile, the annual mean API values were in a small range of 52-53, the air quality levels were gradeⅡ, and P values were 1.06-1.21 less than the slight level (P ≤ 1.31). Total monthly mean PM10, SO₂, NO₂ concentrations, and API and P values were consistently higher in winter and spring than during autumn and summer. The results of a correlation analysis showed that temperature and pressure were the major meteorological factors influencing pollution levels. Pollution sources included industrial coal and straw burning, automobiles exhaust and road dust, fireworks, and dust storms.

A single-tube approach for in vitro diagnostics using diatomaceous earth and optical sensor

Zhao, Fei,Koo, Bonhan,Liu, Huifang,Eun Jin, Choong,Shin, Yong Elsevier 2018 Biosensors & bioelectronics Vol.99 No.-

<P><B>Abstract</B></P> <P>Versatile, simple and efficient sample preparation is desirable for point-of-care testing of emerging diseases such as zoonoses, but current sample preparation assays are insensitive, labour-intensive and time-consuming and require multiple instruments. We developed a single-tube sample preparation approach involving direct pathogen enrichment and extraction from human specimens using diatomaceous earth (DE). Amine-modified DE was used to directly enrich a zoonotic pathogen, <I>Brucella</I>, in a large sample volume. Next, a complex of amine-modified DE and dimethyl suberimidate was used for nucleic acid extraction from the enriched pathogen. Using our single-tube approach, the pathogen can be enriched and extracted within 60min at a level of 1 colony formation unit (CFU) from a 1ml sample volume in the same tube. The performance of this approach is 10–100 times better than that of a commercial kit (10<SUP>2</SUP> to 10<SUP>3</SUP> CFU/ml) but does not require a large centrifuge. Finally, we combined the single-tube approach with a bio-optical sensor for rapid and accurate zoonotic pathogen detection in human urine samples. Using the combination system, <I>Brucella</I> in human urine can be efficiently enriched (~ 8-fold) and the detection limit is enhanced by up to 100 times (1CFU/ml bacteria in urine) compared with the commercial kit. This combined system is fast and highly sensitive and thus represents a promising approach for disease diagnosis in the clinical setting.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A single-tube sample preparation approach involving direct pathogen enrichment and extraction. </LI> <LI> Pathogen can be enriched and extracted within 60min at a level of 1 colony. </LI> <LI> Combination of the single-tube approach with a bio-optical sensor for rapid and accurate zoonotic pathogen detection. </LI> <LI> This system can be efficiently enriched (~ 8-fold) and the detection limit is enhanced by up to 100 times. </LI> </UL> </P>

자동차산업에서 위기경영체제가 구성원의 충성도 및 태도변화에 미치는 영향

박상현(Sang Hyeon Park),유혜방(Huifang Liu),정진섭(Jin Sup Jung) 피터드러커 소사이어티 2021 창조와 혁신 Vol.14 No.2

본 연구는 자동차산업에 있어서, 기업의 위기경영체제 조성이 구성원의 충성도 및 태도변화에 미치는 관계를 실증하고자 하였다. 먼저, 기존 문헌을 기반으로 연구모델을 구축하였으며, 자동차산업에 종사하고 있는 임·직원을 대상으로 설문조사를 실시하였다. 총 203부의 설문지를 분석에 활용하였다. 실증분석의 결과를 보면, 기업의 위기경영체제는 구성원의 충성도에 정(+)의 영향을 미쳤으며, 구성원의 충성도는 태도변화에 정(+)의 영향을 미쳤다. 한편 기업의 위기경영체제는 구성원의 태도변화에 통계적으로 유의한 정(+)의 영향을 미치지 않았다. 또한 구성원의 충성도는 위기경영체제와 태도변화 사이에 완전매개효과를 보여주었는데, 이는 종업원의 충성도가 중요함을 시사하고 있다. 끝으로, 결론 부문에서는 본 연구 결과를 기반으로 다양한 측면의 시사점 및 한계점을 제시하였다. This study sought to demonstrate the relationship of a company s ‘crisis management system’ to its members ‘loyalty’ and ‘the change of attitude’ in the automobile industry. First, a research model was established based on the existing literature, and a survey was conducted on executives and employees engaged in the automobile industry. A total of 203 copies were used for analysis. According to the results of the empirical analysis, the company s ‘crisis management system’ influenced the ‘loyalty’ of its members, and the ‘loyalty’ influenced the ‘change of attitude.’ On the other hand, the company s crisis management system did not have a statistically significant effect on the change of attitude of its members. Members loyalty also showed a complete mediating effect between the crisis management system and the change of attitude, suggesting that employees loyalty is important. Finally, the conclusion section suggests various aspects of implications and limitations based on the results of this research.