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Directed Evolution of Beta-galactosidase from Escherichia coli into Beta-glucuronidase
Xiong, Ai-Sheng,Peng, Ri-He,Zhuang, Jing,Liu, Jin-Ge,Xu, Fang,Cai, Bin,Guo, Zhao-Kui,Qiao, Yu-Shan,Chen, Jian-Min,Zhang, Zhen,Yao, Quan-Hong Korean Society for Biochemistry and Molecular Biol 2007 Journal of biochemistry and molecular biology Vol.40 No.3
In vitro directed evolution through DNA shuffling is a powerful molecular tool for creation of new biological phenotypes. E. coli $\beta$-galactosidase and $\beta$-glucuronidase are widely used, and their biological function, catalytic mechanism, and molecular structures are well characterized. We applied an in vitro directed evolution strategy through DNA shuffling and obtained five mutants named YG6764, YG6768, YG6769, YG6770 and YG6771 after two rounds of DNA shuffling and screening, which exhibited more $\beta$-glucuronidase activity than wild-type $\beta$-galactosidase. These variants had mutations at fourteen nucleic acid sites, resulting in changes in ten amino acids: S193N, T266A, Q267R, V411A, D448G, G466A, L527I, M543I, Q626R and Q951R. We expressed and purified those mutant proteins. Compared to the wild-type protein, five mutant proteins exhibited high $\beta$-glucuronidase activity. The comparison of molecular models of the mutated and wildtype enzymes revealed the relationship between protein function and structural modification.
Single-cell RNA sequencing reveals B cell–related molecular biomarkers for Alzheimer’s disease
Xiong Liu-Lin,Xue Lu-Lu,Du Ruo-Lan,Niu Rui-Ze,Chen Li,Chen Jie,Hu Qiao,Tan Ya-Xin,Shang Hui-Fang,Liu Jia,Yu Chang-Yin,Wang Ting-Hua 생화학분자생물학회 2021 Experimental and molecular medicine Vol.53 No.-
In recent years, biomarkers have been integrated into the diagnostic process and have become increasingly indispensable for obtaining knowledge of the neurodegenerative processes in Alzheimer’s disease (AD). Peripheral blood mononuclear cells (PBMCs) in human blood have been reported to participate in a variety of neurodegenerative activities. Here, a single-cell RNA sequencing analysis of PBMCs from 4 AD patients (2 in the early stage, 2 in the late stage) and 2 normal controls was performed to explore the differential cell subpopulations in PBMCs of AD patients. A significant decrease in B cells was detected in the blood of AD patients. Furthermore, we further examined PBMCs from 43 AD patients and 41 normal subjects by fluorescence activated cell sorting (FACS), and combined with correlation analysis, we found that the reduction in B cells was closely correlated with the patients’ Clinical Dementia Rating (CDR) scores. To confirm the role of B cells in AD progression, functional experiments were performed in early-stage AD mice in which fibrous plaques were beginning to appear; the results demonstrated that B cell depletion in the early stage of AD markedly accelerated and aggravated cognitive dysfunction and augmented the Aβ burden in AD mice. Importantly, the experiments revealed 18 genes that were specifically upregulated and 7 genes that were specifically downregulated in B cells as the disease progressed, and several of these genes exhibited close correlation with AD. These findings identified possible B cell-based AD severity, which are anticipated to be conducive to the clinical identification of AD progression.
Meng Zhang,Xianfeng Qiao,Xiong Peng,Maohua Chen 한국응용곤충학회 2016 Journal of Asia-Pacific Entomology Vol.19 No.2
The bird cherry-oat aphid, Rhopalosiphumpadi (L.) (Hemiptera: Aphididae), is an important pest insect ofwheat, Triticum aestivum (L.), in China. The susceptibility and resistance of wheat cultivars to different populations of R. padi have been rarely reported. In the current study, eight R. padi populations were collected from different provinces of China, the resistance or susceptibility of six wheat cultivars (Emai 596, Emai 18, Emai 251, Zhengmai 98, Xanthus and 35A9B) to these eight populations was tested by the ratio of aphid abundance according the standard method previously reported. The results showed that R. padi populations from Lanzhou of Gansu Province and Taian of Shandong Province showed similar pattern of aphid-host reactions among the six wheat cultivars. Populations from Nanyang of Henan Province and Baicheng of Jilin Province shared a different pattern of reaction. The respective populations from Xianyang of Shaaxi Privince, Beibei of Chongqing Province, Hongtong of Shanxi Province and Baoding of Hebei Province showed unique resistance/susceptible patterns of aphid-host reactions from other populations. The variability in the response to wheat cultivars to R. padi indicated the existence of biotypic variation in populations. The weight of adult aphid survivors of different populations varied on six wheat cultivars, but did not correspond well to the susceptibility and resistance of wheat cultivars. Our findings are important for the sustainable control of R. padi in different wheat growing regions of China.
Wen, Ying,Pan, Xiong-Fei,Huang, Wen-Zhi,Zhao, Zhi-Mei,Wei, Wen-Qiang,Chen, Feng,Lan, Hui,Huang, He,Yang, Chun-Xia,Qiao, You-Lin Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.1
Background: The current study examined health-related quality of life (QoL) for patients with esophageal/gastric cardia precursor lesions or cancer before and after treatment to facilitate improved prevention and treatment. Materials and Methods: Patients with different stages of esophageal/gastric cardia lesions completed two QoL questionnaires, EORTC QLQ-C30 and supplemental QLQ-OES 18, before primary treatment, and at 1, 6 and 12 months after treatment. Results: Fifty-nine patients with precursor lesions, 57 with early stage cancer, and 43 with advanced cancer responded to our survey. Patients with precursor lesions or early stage cancer reported better QoL overall than those with advanced cancer before treatment (p<0.01). Global QoL scores before treatment and at 1 month after treatment were $71{\pm}9$ versus $69{\pm}9$ (p>0.01), $71{\pm}8$ versus $61{\pm}11$ (p<0.01), $67{\pm}11$ versus $62{\pm}9$ (p<0.01) for three stages of lesions. At 6 months after treatment, some QoL measures recovered gradually in precursor lesion and early cancer patients, while some continuously deteriorated in advanced cancer patients. At 12 months, all QoL scores were comparable to baseline for patients with precursor lesions (p>0.01), while global QoL, social, pain, and insomnia scores for early stage and advanced cancer were inferior to corresponding baseline levels (difference between means>5, p<0.01). At this time point, compared with patients with early stage cancer, those with advanced cancer showed worse QoL with all function and most symptom measures (p<0.01). Conclusions: Patients with precursor lesions or early stage esophageal/gastric cardia cancer show better QoL than those with advanced cancer. This indicates that screening, early diagnosis and treatment may improve the QoL for esophageal/gastric cardia cancer patients. Target intervention and counseling should be given by health care providers during treatment and follow-up to facilitate QoL improvement.
The small GTPase CDC42 regulates actin dynamics during porcine oocyte maturation
ZHANG, Yu,WANG, Qiao-Chu,LIU, Jun,XIONG, Bo,CUI, Xiang-Shun,KIM, Nam-Hyung,SUN, Shao-Chen 家畜繁殖硏究所 2017 Journal of Reproduction and Development Vol. No.
<P> The mammalian oocyte undergoes an asymmetric division during meiotic maturation, producing a small polar body and a haploid gamete. This process involves the dynamics of actin filaments, and the guanosine triphosphatase (GTPase) protein superfamily is a major regulator of actin assembly. In the present study, the small GTPase CDC42 was shown to participate in the meiotic maturation of porcine oocytes. Immunofluorescent staining showed that CDC42 was mainly localized at the periphery of the oocytes, and accumulated with microtubules. Deactivation of CDC42 protein activity with the effective inhibitor ML141 caused a decrease in actin distribution in the cortex, which resulted in a failure of polar body extrusion. Moreover, western blot analysis revealed that besides the Cdc42-N-WASP pathway previously reported in mouse oocytes, the expression of ROCK and p-cofilin, two molecules involved in actin dynamics, was also decreased after CDC42 inhibition during porcine oocyte maturation. Thus, our study demonstrates that CDC42 is an indispensable protein during porcine oocyte meiosis, and CDC42 may interact with N-WASP, ROCK, and cofilin in the assembly of actin filaments during porcine oocyte maturation.</P>
Manli Li,Enqi Jin,Zhiyong Qiao,Yuting Xiong 한국섬유공학회 2019 Fibers and polymers Vol.20 No.9
In order to study the effects of molecular structure of carboxyl-containing vinyl monomers on applicationperformance of feather keratin (FK) graft copolymer for warp sizing, three kinds of carboxyl acids, i.e. acrylic acid (AA),methacrylic acid (MAA), and itaconic acid (IA), were grafted onto molecular chains of FK through initiation of K2S2O8-NaHSO3 redox system. FK-g-PAA, FK-g-PMAA, and FK-g-PIA with similar grafting ratios around 31 % were prepared assizing agents. Major sizing properties of the three grafted keratins were studied in terms of water-solubility, apparentviscosity, contact angles of sizing paste on cotton fibers, mechanical properties of sizing film, and adhesion to fibers. It wasfound that sizing properties of the grafted FK strongly depended on molecular structure of the carboxyl-containing vinylmonomers. In view of overall performance of the grafted keratin sizes, the most appropriate carboxyl-containing vinylmonomer should be AA.
Yi-Kang Sun,Xiao-Long Li,Qiao Wang,Bo-Yang Zhou,An-Qi Zhu,Chuan Qin,Le-Hang Guo,Hui-Xiong Xu 대한초음파의학회 2022 ULTRASONOGRAPHY Vol.41 No.2
Purpose: This prospective study explored the value of synchronous tele-ultrasound (US) to aid doctors inexperienced in US with breast US examinations.Methods: In total, 99 patients were enrolled. Two trainee doctors who were inexperienced in US (trainee A [TA] and trainee B [TB]) and one doctor who was an expert in US completed the US examinations sequentially. TA completed the US examinations independently, while TB was instructed by the expert using synchronous tele-US. Subsequently, the expert performed on-site US examinations in person. Separately, they selected the most clinically significant nodule as the target nodule. Consistency with the expert and image quality were compared between TA and TB to evaluate tele-US. Furthermore, TB and the patients evaluated tele-US through questionnaires.Results: TB demonstrated higher consistency with the expert in terms of target nodule selection than TA (93.3% vs. 63.3%, P<0.001). TB achieved good inter-observer agreement (ICC, >0.75) with the expert on five US features (5/9, 55.6%), while TA only did so for one (1/9, 11.1%) (P=0.046). TB’s image quality was higher than TA’s in gray value, time gain compensation, depth, color Doppler adjustment, and the visibility of key information (P=0.018, P<0.001, P<0.001, P=0.033, and P=0.006, respectively). The comprehensive assessment score was higher for TB than for TA (3.96±0.82 vs. 3.09±0.87, P<0.001). Tele-US was helpful in 69.7% of US examinations and had a training effect in 68.0%. Furthermore, 63.6% of patients accepted tele-US and 60.6% were willing to pay.Conclusion: Tele-US can help doctors inexperienced in US to perform breast US examinations.