1.Protection Wei Qi to Prevent The Six Channels Transmission Change
Journal of Zhejiang Chinese Medical University 2014;(4):392-393
[Objective] To discuss application of Protection Wei Qi in the six channels transmission change. [Methods] Study and summarize application of Protection Wei Qi in the six channels transmission change through methods of literature research and comprehensive analysis. [Results] The strength of Wei Qi in the book mostly is a key factor in the development of outcomes in the Six channels Disease.He always takes Protection Wei Qi as a fundamental criterion in the Six channels Disease treatment. Disease in Sanyang, Protect the Wei Qi by Quxie;Disease into Sanyin, Protect the Wei Qi by Fuzheng. [Conclusion] Zhang Zhongjing had great importance to the role of the Wei Qi in clinic. And he ran through Shang Han Lun always by treatment ideas of Protection Wei Qi. The performance of Protection Wei Qi though was particularly prominent in the six channels transmission change.
2.Long noncoding RNA MALAT1 regulates Rac1b expression and is associ-ated with colorectal cancer metastasis
Zihuan YANG ; Xingzhi FENG ; Lekun FANG ; Dandan HUANG ; Jianping WANG
Chinese Journal of Pathophysiology 2015;(8):1417-1421
AIM:ToinvestigatetheroleofMALAT1incolorectalcancermetastasis.METHODS:ThemRNA expression levels of MALAT1 and Rac1b in the tumor and adjacent normal tissues were examined by real-time PCR. MALAT1 was knocked down by siRNA in colorectal cancer cell lines .The expression of Rac1b and the epithelial-mesen-chymal transition markers was examined by Western blot .Cell proliferation was determined by EdU analysis .The effects of MALAT1 on the cell migration and invasion were examined by Transwell assay .RESULTS: The expression of MALAT1 was down-regulated in colorectal cancer .Down-regulation of MALAT1 induced Rac1b overexpression, which in turn in-creased the expression levels of E-cadherin and β-catenin.Furthermore, down-regulation of MALAT1 promoted the cell proliferation, invasion and migration.CONCLUSION:MALAT1 is associated with metastasis of colorectal cancer through regulating the expression of Rac1b and the downstream factors.
3.Synthesis of pyrroloquinoline quinone by recombinant Gluconobacter oxydans.
Runle YE ; Feng LI ; Fan DING ; Zhenhui ZHAO ; Sheng CHEN ; Jianfeng YUAN
Chinese Journal of Biotechnology 2020;36(6):1138-1149
Pyrroloquinoline quinone (PQQ), an important redox enzyme cofactor, has many physiological and biochemical functions, and is widely used in food, medicine, health and agriculture industry. In this study, PQQ production by recombinant Gluconobacter oxydans was investigated. First, to reduce the by-product of acetic acid, the recombinant strain G. oxydans T1 was constructed, in which the pyruvate decarboxylase (GOX1081) was knocked out. Then the pqqABCDE gene cluster and tldD gene were fused under the control of endogenous constitutive promoter P0169, to generate the recombinant strain G. oxydans T2. Finally, the medium composition and fermentation conditions were optimized. The biomass of G. oxydans T1 and G. oxydans T2 were increased by 43.02% and 38.76% respectively, and the PQQ production was 4.82 and 20.5 times higher than that of the wild strain, respectively. Furthermore, the carbon sources and culture conditions of G. oxydans T2 were optimized, resulting in a final PQQ yield of (51.32±0.899 7 mg/L), 345.6 times higher than that of the wild strain. In all, the biomass of G. oxydans and the yield of PQQ can be effectively increased by genetic engineering.
Fermentation
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Gene Knockout Techniques
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Gluconobacter oxydans
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genetics
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metabolism
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Industrial Microbiology
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methods
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Multigene Family
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genetics
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Organisms, Genetically Modified
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PQQ Cofactor
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biosynthesis
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genetics
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Promoter Regions, Genetic
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genetics
4.Molecular engineering of transketolase from Escherichia coli and tartaric semialdehyde biosynthesis.
Jianfeng WANG ; Wenying LI ; Zhenqi XIN ; Wenna FENG ; Xiaomin SUN ; Jianfeng YUAN
Chinese Journal of Biotechnology 2022;38(12):4615-4629
Transketolase (EC 2.2.1.1, TK) is a thiamine diphosphate-dependent enzyme that catalyzes the transfer of a two-carbon hydroxyacetyl unit with reversible C-C bond cleavage and formation. It is widely used in the production of chemicals, drug precursors, and asymmetric synthesis by cascade enzyme catalysis. In this paper, the activity of transketolase TKTA from Escherichia coli K12 on non-phosphorylated substrates was enhanced through site-directed saturation mutation and combined mutation. On this basis, the synthesis of tartaric semialdehyde was explored. The results showed that the optimal reaction temperature and pH of TKTA_M (R358I/H461S/R520Q) were 32 ℃ and 7.0, respectively. The specific activity on d-glyceraldehyde was (6.57±0.14) U/mg, which was 9.25 times higher than that of the wild type ((0.71±0.02) U/mg). Based on the characterization of TKTA_M, tartaric acid semialdehyde was synthesized with 50 mmol/L 5-keto-d-gluconate and 50 mmol/L non-phosphorylated ethanolaldehyde. The final yield of tartaric acid semialdehyde was 3.71 g with a molar conversion rate of 55.34%. Hence, the results may facilitate the preparation of l-(+)-tartaric acid from biomass, and provide an example for transketolase-catalyzed non-phosphorylated substrates.
Escherichia coli/genetics*
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Transketolase/chemistry*
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Tartrates
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Escherichia coli Proteins/genetics*