Elevating Limonene Production in Oleaginous Yeast Yarrowia lipolytica via Genetic Engineering of Limonene Biosynthesis Pathway and Optimization of Medium Composition

Bo-Qian Cheng,Liu-Jing Wei,Yu-Bei Lv,Jun Chen,Qiang Hua 한국생물공학회 2019 Biotechnology and Bioprocess Engineering Vol.24 No.3

As an alternative terpenoid producer, nonconventional oleaginous yeast Yarrowia lipolytica was chosen for limonene production. Y. lipolytica can give high biomass yield and metabolize a broad range of substrates such as glycerol, alkanes, fatty acid, fats, and oils. As previously reported, optimization of limonene synthesis pathway and mevalonate (MVA) pathway leads to the accumulation of 112-fold higher limonene as compared to an initial strain. In this study, we introduced an additional copy of limonene synthesis gene (LS), which resulted in an increase of limonene production. This engineered strain was used to carry out further optimization study. Amongst all the carbon sources tested, the highest level of limonene production was obtained from glycerol, and citrate was selected as an auxiliary carbon source. In fed-batch fermentation with an optimized medium, the engineered strain was found to produce 165.3 mg/L limonene, which corresponds to the highest yield till date for the production of limonene in Y. lipolytica.

Stigma and Efficacy of Zhizhu Kuanzhong Capsules Versus Doxepin in the Treatment of Refractory Functional Dyspepsia: A Randomized Controlled Trial

Qian-Qian Wang,Li Cheng,Bi-Yu Wu,Hong-Yi Qiu,Ping Xu,Bo Wang,Xiu-Juan Yan,Sheng-Liang Chen 대한소화기 기능성질환∙운동학회 2023 Journal of Neurogastroenterology and Motility (JNM Vol.29 No.3

Background/AimsStigma related with antidepressants is prevalent in patients with functional dyspepsia. It affects medication compliance and efficacy. Herbal medicine acquired a deep-rooted cultural identity in relieving dyspeptic symptoms in Asians. The research was designed to compare the effectiveness of Zhizhu Kuanzhong capsules (ZZKZ) versus doxepin hydrochloride (doxepin) on alleviating stigma and medication nonadherence among patients with refractory FD (rFD). MethodsPatients with rFD from February 2021 to February 2022 were randomly allocated to receive either doxepin (n = 56) or ZZKZ (n = 57) combined with omeprazole for 4 weeks. Medication possession ratio (MPR), the disease- and medication-associated stigma were analyzed. The scales were utilized to assess dyspeptic symptoms (Leeds Dyspepsia Questionnaire) and psychological conditions (Generalized Anxiety Disorder Questionnaire and Patient Health Questionnaire). ResultsThe MPR values for ZZKZ were significantly higher than those for doxepin (P < 0.001). The stigma scores decreased in ZZKZ group while increased in doxepin group compared to baseline after treatment. The proportion of patients showing ZZKZ-associated stigma was significantly lower than doxepin-associated stigma (P < 0.001). The MPR values were negatively correlated with post-treatment stigma scores in both groups (P < 0.001). Dyspeptic symptoms and psychological condition were improved in both groups after treatment, with no significant difference on post-treatment Leeds Dyspepsia Questionnaire, Generalized Anxiety Disorder Questionnaire, or Patient Health Questionnaire scores between 2 groups. ConclusionZZKZ is superior to doxepin in alleviating stigma and medication non-adherence, with comparable efficacy in improving dyspeptic symptoms and psychological condition of patients with rFD.

MicroRNA-206 Reduces Osteosarcoma Cell Malignancy In Vitro by Targeting the PAX3-MET Axis

Qian-Rong Deng,Fang-Biao Zhan,Xian-Wei Zhang,Shi-Long Feng,Jun Cheng,You Zhang,Bo Li,Li-Zhong Xie 연세대학교의과대학 2019 Yonsei medical journal Vol.60 No.2

Purpose: This study was undertaken to explore how miR-206 represses osteosarcoma (OS) development. Materials and Methods: Expression levels of miR-206, PAX3, and MET mRNA were explored in paired OS and adjacent tissuespecimens. A patient-derived OS cell line was established. miR-206 overexpression and knockdown were achieved by lentiviraltransduction. PAX3 and MET overexpression were achieved by plasmid transfection. Treatment with hepatocyte growth factor(HGF) was utilized to activate c-Met receptor. Associations between miR-206 and PAX3 or MET mRNA in OS cells were verifiedby AGO2-RNA immunoprecipitation assay and miRNA pulldown assay. OS cell malignancy was evaluated in vitro by cell proliferation,metastasis, and apoptosis assays. PAX3 and MET gene expression in OS cells was assayed by RT-qPCR and Western blot. Activation of PI3K-AKT and MAPK-ERK in OS cells were assayed by evaluating Akt1 Ser473 phosphorylation and total threoninephosphorylation of Erk1/2, respectively. Results: Expression levels of miR-206 were significantly decreased in OS tissue specimens, compared to adjacent counterparts,and were inversely correlated with expression of PAX3 and MET mRNA. miR-206 directly interacted with PAX3 and MET mRNAin OS cells. miR-206 overexpression significantly reduced PAX3 and MET gene expression in OS cells in vitro, resulting in significantdecreases in Akt1 and Erk1/2 activation, cell proliferation, and metastasis, as well as increases in cell apoptosis, while miR-206 knockdown showed the opposite effects. The effects of miR-206 overexpression on OS cells were reversed by PAX3 or METoverexpression, but only partially attenuated by HGF treatment. Conclusion: miR-206 reduces OS cell malignancy in vitro by targeting PAX3 and MET gene expression.

Low-loss carrier-stored IGBT with p-type Schottky diode-clamped shielding layer

Yi, Bo,Zhao, Qing,Zhang, Qian,Cheng, JunJi,Huang, HaiMeng,Pan, YiLan,Hu, XiaoRan,Xiang, Yong The Korean Institute of Power Electronics 2021 JOURNAL OF POWER ELECTRONICS Vol.21 No.8

A novel carrier-stored trench bipolar transistor (CSTBT) with heavily doped carrier-stored layer (CSL) is proposed and investigated by TCAD tools. The voltage of CSL is shielded by a buried p-type layer (P-bury) whose potential is clamped by a p-type Schottky Barrier Diode (pSBD) in series-connection with a PN diode. Hence, the CSL can be heavily doped, and the trade-off between on-state voltage drop (V_on) and turn-off loss (E_off) is substantially improved. Compared with that of a conventional CSTBT with floating P-base (FP-CSTBT), the E_off of the proposed CSTBT is reduced by 27.9% at V_on=1.1 V. Owing to the shielding effect of the P-bury layer, the saturation current density of the proposed CSTBT is reduced by 52% compared with that of the FP-CSTBT. Consequently, significantly enlarged short-circuit safe operation area is obtained, and the short-circuit withstand time (t_sc) is increased to 12.8 s at ultra-low V_on (~1.1 V).

Statistics-based Optimization of Extracellular Polysaccharide Production from Hirsutella sinensis using a Fermentation Process and In vitro Immunomodulatory Activity

Liang He,Jun-Wen Cheng,Yan-Bin Wang,Hai-Bo Li,Hua Qian,Wei-Qi Li,Xue-Yong Ren 한국식품과학회 2014 Food Science and Biotechnology Vol.23 No.2

A 3 factor, 3 level Box-Behnken factorialdesign combined with response surface methodology wasused to optimize the fermentation process for production ofextracellular polysaccharide (EPS) from Hirsutella sinensis. The 3 independent variables were temperature (X1), initialpH (X2), and the glucose concentration (X3). Experimentaldata were fitted to a 2nd order polynomial equation usingmultiple regression. The optimal fermentation conditionsof the production of EPS were a temperature of 18.21oC,an initial pH of 5.81, and a glucose concentration of 7.39g/L. The maximum predicted EPS production of 2.41 g/Lwas close to the actual experimental EPS yield (2.42±0.038%), demonstrating the model validity. EPS from H. sinensis had a direct in vitro immuno-stimulating activityusing murine macrophage RAW264.7 cells, and stimulatedrelease of several major cytokines (IL-1β, NF-κB, TNF-α,and iNOS) in a dose-dependent manner.

Selection signature reveals genes associated with susceptibility loci affecting respiratory disease due to pleiotropic and hitchhiking effect in Chinese indigenous pigs

Xu, Zhong,Sun, Hao,Zhang, Zhe,Zhang, Cheng-Yue,Zhao, Qing-bo,Xiao, Qian,Olasege, Babatunde Shittu,Ma, Pei-Pei,Zhang, Xiang-Zhe,Wang, Qi-Shan,Pan, Yu-Chun Asian Australasian Association of Animal Productio 2020 Animal Bioscience Vol.33 No.2

Objective: Porcine respiratory disease is one of the most important health problems causing significant economic losses. To understand the genetic basis for susceptibility to swine enzootic pneumonia (EP) in pigs, we detected 102,809 single nucleotide polymorphisms in a total of 249 individuals based on genome-wide sequencing data. Methods: Genome comparison of susceptibility to swine EP in three pig breeds (Jinhua, Erhualian, and Meishan) with two western lines that are considered more resistant (Duroc and Landrace) using cross-population extended haplotype homozygosity and F-statistic (F_ST) statistical approaches identified 691 positively selected genes. Based on quantitative trait loci, gene ontology terms and literature search, we selected 14 candidate genes that have convincible biological functions associated with swine EP or human asthma. Results: Most of these genes were tested by several methods including transcription analysis and candidate genes association study. Among these genes: cytochrome P450 1A1 and catenin beta 1 (CTNNB1) are involved in fertility; transforming growth factor beta receptor 3 plays a role in meat quality traits; Wnt family member 2, CTNNB1 and transcription factor 7 take part in adipogenesis and fat deposition simultaneously; plasminogen activator, urokinase receptor (completely linked to AXL receptor tyrosine kinase, r² = 1) plays an essential role in the successful ovulation of matured oocytes in pigs; colipase like 2 (strongly linked to SAM pointed domain containing ETS transcription factor, r² = 0.848) is involved in male fertility. Conclusion: These adverse genes susceptible to swine EP may be selected while selecting for economic traits (especially reproduction traits) due to pleiotropic and hitchhiking effect of linked genes. Our study provided a completely new point of view to understand the genetic basis for susceptibility or resistance to swine EP in pigs thereby, provides insight for designing sustainable breed selection programs. Finally, the candidate genes are crucial due to their potential roles in respiratory diseases in a large number of species, including human.