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To develop a novel glucose-lowering biomedicine with potential benefits in the treatment of type 2 diabetes, we used the 10rolGLP-1 gene previously constructed in our laboratory and the CRISPR/Cas9 genome editing technique to create an engineered Saccharomyces cerevisiae strain. The gRNA expression vector pYES2-gRNA, the donor vector pNK1-L-PGK-10rolGLP-1-R and the Cas9 expression vector pGADT7-Cas9 were constructed and co-transformed into S. cerevisiae INVSc1 strain, with the PGK-10rolGLP-1 expressing unit specifically knocked in through homologous recombination. Finally, an S. cerevisiae strain highly expressing the 10rolGLP-1 with glucose-lowering activity was obtained. SDS-PAGE and Western blotting results confirmed that two recombinant strains of S. cerevisiae stably expressed the 10rolGLP-1 and exhibited the desired glucose-lowering property when orally administered to mice. Hypoglycemic experiment results showed that the recombinant hypoglycemic S. cerevisiae strain offered a highly hypoglycemic effect on the diabetic mouse model, and the blood glucose decline was adagio, which can avoid the dangerous consequences caused by rapid decline in blood glucose. Moreover, the body weight and other symptoms such as polyuria also improved significantly, indicating that the orally hypoglycemic S. cerevisiae strain that we constructed may develop into an effective, safe, economic, practical and ideal functional food for type 2 diabetes mellitus treatment.
Asunto(s)
Ratones , Animales , Saccharomyces cerevisiae/metabolismo , Sistemas CRISPR-Cas , Glucosa/metabolismo , Glucemia/metabolismo , Diabetes Mellitus Tipo 2/terapia , Hipoglucemiantes/metabolismoRESUMEN
Santalene and santalol are the main components of valuable perfume sandalwood essential oil, and have good antibacterial, anti-oxidation and anti-tumor activities. Commercial sandalwood essential oil is mainly extracted from sandalwood tree that grows slowly and is difficult to cultivate. In addition, the extraction recovery of sandalwood essential oil from sandalwood tree is too low to meet the market demand. These factors make sandalwood essential oil expensive. An option is to use genetic engineering and molecular biological methods to heterologously express related synthase of santalene and santalol in microbial host. In this paper, the biosynthesis progress of santalene and santalol synthase, as well as the optimization of mevalonate metabolic pathways in the hosts are summarized. Furthermore, the strategies of applying protein engineering technology to carry out orthomutation of santalene synthase were also discussed, to provide reference for the optimal biosynthesis of santalene and santalol.
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Objective:To observe the clinical efficacy of warm needling moxibustion plus tuina in treating knee osteoarthritis (KOA) due to cold-dampness blocking collaterals. Methods:Forty-eight KOA patients were randomized into 2 groups by their visiting sequence, 24 cases in each group. The treatment group was intervened by warm needling moxibustion plus tuina, while the control group was treated with acupuncture plus tuina. The two groups were both treated once a day, 30 min for each session, 10 d as a treatment course, totally for 3 treatment courses. The visual analogue scale (VAS) and Lysholm knee scoring scale (LKSS) were observed before and after the treatment to evaluate the clinical efficacy. Results:After 3 treatment courses, the VAS and LKSS scores were significantly changed in the two groups (P Conclusion:Warm needling moxibustion plus tuina can produce a more significant efficacy than acupuncture plus tuina in treating KOA due to cold-dampness blocking collaterals.
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Objective To investigate the medical and pharmaceutical knowledge of patients with chronic diseases and analyse the influence factors of rational administration in patients ,to provid data to support the establishment of pharmaceutical service mode . Methods 386 cases of patients with chronic diseases were asked to finish the questionnaires for the medical and pharmaceutical knowledge ,and factors affecting the rational drug were analyzed by single factor and multiple factors Logistic regression analysis . Results Among the 386 patients ,cardiovascular and celebralvascular disease ratio was the highest(53 .3% ) ,followed by respiratory system diseases(13 .8% ) and the musculoskeletal system diseases (11 .50% );The averaged score of 386 patients was 1 .76 ± 0 .78 , medication knowledge was at a general level;single factor analysis results showed that there was significant difference(P<0 .05) between rational drug-use and abuse of drugs among patients in number ,form of payment ,marital status ,income ,education level , taking drug knowledge lectures ,combined treatment .Multivariate Logistic regression analysis showed that education level ,partici-pation in lectures ,drug combination ,disease species had a significant impact on the rational drug use among patients with chronic disease(P<0 .05) .Conclusion The pharmaceutical knowledge that patients with chronic disease mastered is unsatisfactory ;and unreasonable behavior of medication is common scence .Education level ,participation in lectures ,drug combination ,the number of diseases have great influence on the rational use of drugs in patients with chronic diseases .A kind of effective pharmaceutical service mode should be established for patients with chronic diseases by clinical pharmacists .This is a very meaningful work for rational ad-ministration .
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Objective To measure 222Rn and 220Rn concentrations in the underground non-uranium mines in Shandong Province,and to estimate the annual effective dose to the miners.Methods Concentrations of 222Rn and 220Rn in selected gold,iron,coal and clay mines were determined by passive time-integrating detectors with CR-39.Activity concentrations of 226Ra,232Th and 40K were determined using gamma spectrometry equipped with HPGe detector.Results The average concentrations of 220Rn in the gold,iron,coal and clay mines were estimated to be 1200,280,120 and 40 Bq/m3,respectively.The activity concentrations of 226Ra,232Th,40K in the ores for gold,iron,coal and clay mines ranged the same as the soil in China.The annual effective dose due to radon exposure in gold and iron mine was 7.70 mSv and 1.74 mSv,respectively.The annual doses received by miners in the coal and clay mines were lower than 1 mSv.Conclusions Underground miners in some gold and iron mines should be treated as workers occupationally exposed to ionizing radiation.The measurement such as increasing ventilation should be implemented to reduce underground radon concentration in these mines.