Improvement of FK506 Production by Selection of 4-Aminobutyric Acid-tolerant Mutant and Optimization of its Fermentation Using Response Surface Methodology

Tianyi Yang,Jiaojiao Li,Liangzhi Li,Haiyang Zhang,Jing Ma,Zhi Chen,Cuiying Hu,Xin Ju,Jiaolong Fu 한국응용생명화학회 2014 Applied Biological Chemistry (Appl Biol Chem) Vol.57 No.6

Tacrolimus (FK506), a 23-membered polyketidemacrolide with immunosuppressant activity, can be produced byStreptomyces tsukubaensis. We studied a variety of mutant strainsof S. tsukubaensis for the microbial production of FK506. Thebest strain (CZ-19) was obtained from the parent strain LLZ-1 bynitrosoguanidine mutation and 4-Aminobutyric acid (FK506precursor structure analogs) adaption. In the shake-flask experiments,titer of FK506 by CZ-19 was 532.44 mg/L, increased by 65.13%compared to that of the parent strain. Through single factorexperiments and response surface methodology, we furtheroptimized the medium for improved FK506 production by CZ-19in shake flask culture. The optimal medium for enhanced FK506production was as follows: 17.19 g/L corn starch, 21.78 g/Lglucose, 8.06 g/L peptone, and 18.98 mg/L 4-aminobutyric acid. The predicted FK506 titer was 906.49 mg/L, and the experimentaldata confirmed the validity of the model. The present studydemonstrates that S. tsukubaensis CZ-19 is a promising strain forindustrial production of FK506.

Improvement of FK506 Production by Selection of 4-Aminobutyric Acid-tolerant Mutant and Optimization of its Fermentation Using Response Surface Methodology

Yang, Tianyi,Li, Jiaojiao,Li, Liangzhi,Zhang, Haiyang,Ma, Jing,Chen, Zhi,Hu, Cuiying,Ju, Xin,Fu, Jiaolong 한국응용생명화학회 2014 Applied Biological Chemistry (Appl Biol Chem) Vol.57 No.6

Tacrolimus (FK506), a 23-membered polyketide macrolide with immunosuppressant activity, can be produced by Streptomyces tsukubaensis. We studied a variety of mutant strains of S. tsukubaensis for the microbial production of FK506. The best strain (CZ-19) was obtained from the parent strain LLZ-1 by nitrosoguanidine mutation and 4-Aminobutyric acid (FK506 precursor structure analogs) adaption. In the shake-flask experiments, titer of FK506 by CZ-19 was 532.44 mg/L, increased by 65.13% compared to that of the parent strain. Through single factor experiments and response surface methodology, we further optimized the medium for improved FK506 production by CZ-19 in shake flask culture. The optimal medium for enhanced FK506 production was as follows: 17.19 g/L corn starch, 21.78 g/L glucose, 8.06 g/L peptone, and 18.98 mg/L 4-aminobutyric acid. The predicted FK506 titer was 906.49 mg/L, and the experimental data confirmed the validity of the model. The present study demonstrates that S. tsukubaensis CZ-19 is a promising strain for industrial production of FK506.

디지털 클로딩 기술 기반 가상착의 업사이클링 패션디자인

천탠이 ( Tianyi Chen ),양은경 ( Eun Kyoung Yang ),이연희 ( Younhee Lee ) 복식문화학회 2021 服飾文化硏究 Vol.29 No.3

The purpose of this study is to develop up-cycling fashion design methods centered on discarded denim material for the study of original up-cycling design methods. Up-cycling fashion design work was developed using digital clothing technology. This is a recent hot topic among sustainable fashion design methods. Up-cycling fashion design expression methods (categorized as dismantlement, collages, dépaysement, grafting, weaving, and tearing) were centered on design methods. These methods create various three-dimensional modeling effects in planar forms, whereby five pieces can be applied to the fabric and digitally produced. The results are as follows: First, the use of discarded denim fabric for the development of up-cycling fashion design pieces enabled the recycling of existing resources, provided solutions to environmental pollution problems, and provided expansion opportunities for design processes for sustainable fashion products that expand the design value of denim products and their utility. Second, new eco-friendly fashion designs that attempt to achieve diversity in modern fashion trends could be presented through formative contemporary fashion produced by up-cycling work products. Third, up-cycling fashion design work is expected to provide opportunities for eco-friendly fashion design methods. This will expand the value of sustainable fashion design by recycling simple waste materials through the use of three-dimensional digital clothing technology and further through the presentation of expanded life cycles that extend product planning, production, and life cycles.

Construction of an Efficient Mutant Strain of Trichosporonoides oedocephalis with HOG1 Gene Deletion for Production of Erythritol

( Liangzhi Li ),( Tianyi Yang ),( Weiqiang Guo ),( Xin Ju ),( Cuiying Hu ),( Bingyu Tang ),( Jiaolong Fu ),( Jingsheng Gu ),( Haiyang Zhang ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.4

The mitogen-activated protein kinase HOG1 (high-osmolarity glycerol response pathway) plays a crucial role in the response of yeast to hyperosmotic shock. Trichosporonoides oedocephalis produces large amounts of polyols (e.g., erythritol and glycerol) in a culture medium. However, the effects of HOG1 gene knockout and environmental stress on the production of these polyols have not yet been studied. In this study, a To-HOG1 null mutation was constructed in T. oedocephalis using the loxP-Kan-loxP/Cre system as replacement of the targeted genes, and the resultant mutants showed much smaller colonies than the wild-type controls. Interestingly, compared with the wild-type strains, the results of shake-flask culture showed that To-HOG1 null mutation increased erythritol production by 1.44-fold while decreasing glycerol production by 71.23%. In addition, this study investigated the effects of citric acid stress on the T. oedocephalis HOG1 null mutants and the wild-type strain. When the supplementation of citric acid in the fermentation medium was controlled at 0.3% (w/v), the concentration of erythritol produced from the wild-type and To-HOG1 knockout mutant strains improved by 18.21% and 21.65%, respectively.

Hepatotoxic mechanism of diclofenac sodium on broiler chicken revealed by iTRAQ-based proteomics analysis

Chuanxi Sun,Tianyi Zhu,Yuwei Zhu,Bing Li,Jiaming Zhang,Yixin Liu,Changning Juan,Shifa Yang,Zengcheng Zhao,Renzhong Wan,Shuqian Lin,Bin Yin 대한수의학회 2022 Journal of Veterinary Science Vol.23 No.4

Background: At the therapeutic doses, diclofenac sodium (DFS) has few toxic side effects on mammals. On the other hand, DFS exhibits potent toxicity against birds and the mechanisms remain ambiguous. Objectives: This paper was designed to probe the toxicity of DFS exposure on the hepatic proteome of broiler chickens. Methods: Twenty 30-day-old broiler chickens were randomized evenly into two groups (n = 10). DFS was administered orally at 10 mg/kg body weight in group A, while the chickens in group B were perfused with saline as a control. Histopathological observations, serum biochemical examinations, and quantitative real-time polymerase chain reaction were performed to assess the liver injury induced by DFS. Proteomics analysis of the liver samples was conducted using isobaric tags for relative and absolute quantification (iTRAQ) technology. Results: Ultimately, 201 differentially expressed proteins (DEPs) were obtained, of which 47 were up regulated, and 154 were down regulated. The Gene Ontology classification and Kyoto Encyclopedia of Genes and Genomes pathway analysis were conducted to screen target DEPs associated with DFS hepatotoxicity. The regulatory relationships between DEPs and signaling pathways were embodied via a protein-protein interaction network. The results showed that the DEPs enriched in multiple pathways, which might be related to the hepatotoxicity of DFS, were “protein processing in endoplasmic reticulum,” “retinol metabolism,” and “glycine, serine, and threonine metabolism.” Conclusions: The hepatotoxicity of DFS on broiler chickens might be achieved by inducing the apoptosis of hepatocytes and affecting the metabolism of retinol and purine. The present study could provide molecular insights into the hepatotoxicity of DFS on broiler chickens.

Outcomes of Anti-CD19 CAR-T Treatment of Pediatric B-ALL with Bone Marrow and Extramedullary Relapse

Xinyu Wan,Xiaomin Yang,Fan Yang,Tianyi Wang,Lixia Ding,Lili Song,Yan Miao,Xiang Wang,Yani Ma,Chengjuan Luo,Jingyan Tang,Longjun Gu,Jing Chen,Yanjing Tang,Jun Lu,Benshang Li 대한암학회 2022 Cancer Research and Treatment Vol.54 No.3

PurposeAnti-CD19 chimeric antigen receptor T-cell immunotherapy (19CAR-T) has achieved impressive clinical results in adult and pediatric relapsed/refractory (r/r) B-lineage acute lymphoblastic leukemia (B-ALL). However, the application and effect of CAR-T therapy in B-ALL patients with extramedullary relapse are rarely issued even disqualified in some clinical trials. Here, we examined the efficacy of 19CAR-T in patients with both bone marrow and extramedullary involvement.Materials and MethodsCAR-T cells were generated by transfection of primary human T lymphocytes with a lentiviral vector expressing anti-CD19 single chain antibody fragments (scFvs) with the cytoplasmic domains of 4-1BB and CD3ζ, and used to infuse patients diagnosed as having r/r B-ALL with extramedullary origination. Clinical responses were evaluated by the use of bone marrow aspiration, imaging, and flow cytometry. ResultsEight patients received 19CAR-T infusion and all attained complete remission (CR). Only one patient was bridged to hematopoietic stem cell transplantation (HSCT). Although three patients relapsed after infusion, they received 19/22CAR-T infusion sequentially and attained a second remission. To date, five patients are in continuous CR and all eight patients are still alive. The mean follow-up time was 21.9 months, while the 24-month estimated event-free survival is 51.4%. Conclusion19CAR-T therapy can lead to clinical remission for extramedullary relapsed pediatric B-ALL patients. However, the problem of CD19+ relapses after CAR-T remained to be solved. For patients relapsing after CAR-T, a second CAR-T therapy creates another opportunity for remission for subsequent HSCT.

Spatial correlation of aerodynamic forces on 5:1 rectangular cylinder in different VIV stages

Yongfu Lei,Yanguo Sun,Tianyi Zhang,Xiongwei Yang,Ming-shui Li 한국풍공학회 2022 Wind and Structures, An International Journal (WAS Vol.34 No.1

To better understand the vortex-induced vibration (VIV) characteristics of a 5:1 rectangular cylinder, the distribution of aerodynamic force and the non-dimensional power spectral density (PSD) of fluctuating pressure on the side surface were studied in different VIV development stages, and their differences in the stationary state and vibration stages were analyzed. The spanwise and streamwise correlations of surface pressures were studied, and the flow field structure partitions on the side surface were defined based on the streamwise correlation analysis. The results show that the variation tendencies of mean and root mean square (RMS) pressure coefficients are similar in different VIV development stages. The RMS values during amplitude growth are larger than those at peak amplitude, and the smallest RMS values are observed in the stationary state. The spanwise correlation coefficients of aerodynamic lifts increase with increase of the peak amplitude. However, for the lock-in region, the maximum spanwise correlation coefficient for aerodynamic lifts occurs in the VIV rising stage rather than in the peak amplitude stage, probably due to the interaction of vortex shedding force (VSF) and self-excited force (SEF). The streamwise correlation results show that the demarcation point positions between the recirculation region and the main vortex region remain almost constant in different VIV development stages, and the reattachment points gradually move to the tailing edge with increasing amplitude. This study provides a reference to estimate the demarcation point and reattachment point positions through streamwise correlation and phase angle analysis from wind tunnel tests.

Hepatotoxic mechanism of diclofenac sodium on broiler chicken revealed by iTRAQ-based proteomics analysis

Sun, Chuanxi,Zhu, Tianyi,Zhu, Yuwei,Li, Bing,Zhang, Jiaming,Liu, Yixin,Juan, Changning,Yang, Shifa,Zhao, Zengcheng,Wan, Renzhong,Lin, Shuqian,Yin, Bin 대한수의학회 2022 Journal of Veterinary Science Vol.23 No.3

Background: At the therapeutic doses, diclofenac sodium (DFS) has few toxic side effects on mammals. On the other hand, DFS exhibits potent toxicity against birds and the mechanisms remain ambiguous. Objectives: This paper was designed to probe the toxicity of DFS exposure on the hepatic proteome of broiler chickens. Methods: Twenty 30-day-old broiler chickens were randomized evenly into two groups (n = 10). DFS was administered orally at 10mg/kg body weight in group A, while the chickens in group B were perfused with saline as a control. Histopathological observations, serum biochemical examinations, and quantitative real-time polymerase chain reaction were performed to assess the liver injury induced by DFS. Proteomics analysis of the liver samples was conducted using isobaric tags for relative and absolute quantification (iTRAQ) technology. Results: Ultimately, 201 differentially expressed proteins (DEPs) were obtained, of which 47 were up regulated, and 154 were down regulated. The Gene Ontology classification and Kyoto Encyclopedia of Genes and Genomes pathway analysis were conducted to screen target DEPs associated with DFS hepatotoxicity. The regulatory relationships between DEPs and signaling pathways were embodied via a protein-protein interaction network. The results showed that the DEPs enriched in multiple pathways, which might be related to the hepatotoxicity of DFS, were "protein processing in endoplasmic reticulum," "retinol metabolism," and "glycine, serine, and threonine metabolism." Conclusions: The hepatotoxicity of DFS on broiler chickens might be achieved by inducing the apoptosis of hepatocytes and affecting the metabolism of retinol and purine. The present study could provide molecular insights into the hepatotoxicity of DFS on broiler chickens.

Ratio of PM2.5 to PM10 Mass Concentrations in Beijing and Relationships with Pollution from the North China Plain

Zhigang Li,Xin Yang,Chuanfeng Zhao,Tianyi Fan 한국기상학회 2021 Asia-Pacific Journal of Atmospheric Sciences Vol.57 No.3

Five years of PM2.5/PM10 ratios and their potential relationships with wind and relative humidity (RH) were analyzed for three areas in Beijing (northwestern mountainous area, urban area and southern suburbs), and these values were compared with those of five other cities, including Tianjin, Dalian, Hangzhou, Shanghai and Guangzhou. In the past five years, both PM2.5 and PM10 mass concentrations have decreased over the three Beijing regions, particularly during winters. The PM2.5/PM10 ratios have distinct seasonal characteristics, with more frequent high ratios in winter than in other seasons. The high ratio frequency decrease is most evident in winter and in the southern Beijing suburbs. This fine particle proportion decrease is related to air pollution control policies, including the national project ‘Coal to Gas’. HighPM2.5/PM10 ratios are linked to heavy pollution levels and low wind speeds, indicating the importance of PM2.5 accumulation during pollution events in Beijing. The higher PM2.5/PM10 ratios in Beijing are also closely related to southerly winds and high humidity, indicating the contribution of anthropogenic pollution transported from the south. Due to similar geographic environments, Tianjin is similar to Beijing in terms of the frequency distribution characteristics of the PM2.5/PM10 ratios. The coastal city of Dalian is further north and not similar to Beijing or Tianjin, owing to a sea breeze influence. Different from Beijing and Tianjin, the southern cities of Hangzhou, Shanghai and Guangzhou show almost no change in the PM2.5/PM10 ratio frequency distribution with increases in wind speed and relative humidity.

Live Imaging of Endogenous PSD-95 Using ENABLED: A Conditional Strategy to Fluorescently Label Endogenous Proteins

Fortin, Dale A.,Tillo, Shane E.,Yang, Guang,Rah, Jong-Cheol,Melander, Joshua B.,Bai, Suxia,Soler-Cedeñ,o, Omar,Qin, Maozhen,Zemelman, Boris V.,Guo, Caiying,Mao, Tianyi,Zhong, Haining Society for Neuroscience 2014 The Journal of neuroscience Vol.34 No.50

<P>Stoichiometric labeling of endogenous synaptic proteins for high-contrast live-cell imaging in brain tissue remains challenging. Here, we describe a conditional mouse genetic strategy termed endogenous labeling via exon duplication (ENABLED), which can be used to fluorescently label endogenous proteins with near ideal properties in all neurons, a sparse subset of neurons, or specific neuronal subtypes. We used this method to label the postsynaptic density protein PSD-95 with mVenus without overexpression side effects. We demonstrated that mVenus-tagged PSD-95 is functionally equivalent to wild-type PSD-95 and that PSD-95 is present in nearly all dendritic spines in CA1 neurons. Within spines, while PSD-95 exhibited low mobility under basal conditions, its levels could be regulated by chronic changes in neuronal activity. Notably, labeled PSD-95 also allowed us to visualize and unambiguously examine otherwise-unidentifiable excitatory shaft synapses in aspiny neurons, such as parvalbumin-positive interneurons and dopaminergic neurons. Our results demonstrate that the ENABLED strategy provides a valuable new approach to study the dynamics of endogenous synaptic proteins <I>in vivo</I>.</P>