To improve the expression level and catalytic efficiency of (R)-carbonyl reductase from Candida parapsilosis in Escherichia coli, we optimized the mRNA secondary structure of (R)-carbonyl reductase gene in translation initiation region (from +1 to +78), and constructed the corresponding variant. The formation of hairpin structure was significantly reduced and the Gibbs free energy was dramatically decreased from -9.5 kcal/mol to -5.0 kcal/mol after optimization. As a result, the expression level of (R)-carbonyl reductase in the variant was increased by 4-5 times and its specific activity in cell-free extract was enhanced by 61.9% compared to the wild-type strain. When using the whole cells as catalyst and 2-hydroxyacetophenone as substrate with a high concentration of 5.0 g/L, the variant showed excellent performance to give (R)-1-phenyl-1, 2-ethanediol with optical purity of 93.1% enantiomeric excess and a yield of 81.8%, which were increased by 27.5% and 40.5% respectively than those of the wild-type. In conclusion, the optimization of mRNA secondary structure in translation initiation region can overcome the steric hindrance of translation startup, promote translation smoothly to acquire high expression of target protein, and favor protein folding correctly to efficiently improve the enzyme specific activity and biotransformation function.
Alcohol Oxidoreductases ; biosynthesis ; chemistry ; genetics ; Base Sequence ; Biocatalysis ; Candida ; enzymology ; Catalysis ; Escherichia coli ; genetics ; metabolism ; Molecular Sequence Data ; Mutant Proteins ; genetics ; Nucleic Acid Conformation ; Peptide Chain Initiation, Translational ; RNA, Messenger ; chemistry ; Recombinant Proteins ; biosynthesis ; genetics ; Stereoisomerism

OBJECTIVE：To estab lish“TCM Data Analysis Platform Based on Strategy Pattern ”（TCMDP），and to analyze the medication rules of TCM prescription against primary hepatic carcinoma （PHC），so as to provide reference for TCM prescription screening. METHODS ：According to the idea of strategy pattern ，TCMDP software was developed ，and the frequency statistics，association，clustering，classification and other methods and the different algorithms covered were encapsulated. Effective TCM prescriptions against PHC were collected and screened from 4 tertiary-grade class-A hospitals in Nanchang city and CNKI ， PubMed. A database of prescriptions for PHC （PDOPHC）was established ，and TCM prescriptions in the data base were put into TCMDP. Clustering algorithm modified by CMC-DD ，modified WD-Get Rules algorithm after merging depth and width search and frequency statistics were used to analyze the distribution of prescription syndrome. Taking phlegm-blood stasis syndrome type （TYPHC）as example ，data mining was carried out on drug efficacy category ，four properties ，five flavors and channel tropism ， drug property combination ，key couplet medicines and key drug combination. RESULTS ：A total of 907 TCM prescriptions against PHC were collected ，mainly involving 10 syndrome types ，such as phlegm-blood stasis syndrome ，deficiency of the vital essential and blood stasis syndrome and liver and spleen blood stasis syndrome. TCM related to TYPHC commonly used in pre- scriptions were tonifying deficiency medicine ， phlegm-re- solving medicine ，promoting blood circulation and removing blood stasi s medicine ，etc．Four properties were mostly cold ，warm and calm ；the five flavors were most ly swe et，bitter and pungent ；channel tropism were mostly spleen ，liver，stomach， lung，heart and kidney ；drug combination were mostly warming-sweet-spleen ，warming-pungent-spleen and cold-bitter-liver ，etc.； there were 36 couplet medicines with frequency ≥30 times；there were 31 strong association rules and 8 key drug combinations in the prescriptions. WD-Get Rules results showed that in the TCM prescriptions against PHC ，Atractylodes macrocephala ，Codonopsis pilosula，Poria cocos and Glycyrrhiza uralensis were in the key position. They were often combined with many kinds of drugs ， which were related to the effect of Sijunzi decoction on strengthening Qi and tonifying spleen. CMC-DD results showed that therapy for TYPHC focused on strengthening the spleen and stomach ，replenishing Qi and blood ；and at the same time ，it treated TYPHC with phlegm-blood stasis syndrome from multiple angles ，such as soothing the liver and regulating Qi ，eliminating phlegm ，clearing heat and detoxification ，promoting blood circulation and nourishing Yin. CONCLUSIONS ：Clinical treatment of TYPHC mainly adopts the methods of invigorating the spleen and replenishing Qi ，soothing the liver and regulating Qi ，promoting blood circulation and removing blood stasis ，which conforms to the medication principle of “strengthening the body and eliminating pathogenic factors”. The application of TCMDP can realize the data mining and analysis of TCM in multi-level and multi-directional way ， which provides a new and effective way for the mining and research of compatibility rules of TCM prescriptions.

Objective To investigate the effect and mechanism of Feixin Decoction(Astragali Radix,Pericae Semen,Carthami Flos,Descurainiae Semen Lepidii Semen,Paeoniae Radix Rubra,etc.)on monocrotaline-induced pulmonary arterial hypertension(PAH)rats based on peroxisome proliferator-activated receptor-γ/nuclear factor-κB(PPAR-γ/NF-κB)signaling pathway.Methods Forty-eight male SD rats were randomly divided into normal group,model group,Sildenafil group(0.025 g·kg-1)and low-,medium-and high-dose of Feixin Decoction groups(11.7,23.4,46.8 g·kg-1).PAH rat model was established by single intraperitoneal injection of monocrotaline solution(60 mg·kg-1).After 1 hour of modeling,the rats were given intragastric administration once a day for 28 days.Hemodynamic and echocardiographic parameters including right ventricular systolic pressure(RVSP),mean pulmonary artery pressure(mPAP),right ventricular hypertrophy index(RVHI),pulmonary artery acceleration time(PAAT),pulmonary artery ejection time(PET),tricuspid annular plane systolic excursion(TAPSE),right ventricular internal diameter(RVIDd)and right ventricular anterior wall thickness(RVAWT)were measured in each group.The pathological changes of pulmonary arterioles were observed by HE staining.The expression level of α-smooth muscle actin(α-SMA)in rat pulmonary artery was detected by immunofluorescence.The levels of plasma interleukin-1β(IL-1β),IL-6 and tumor necrosis factor-α(TNF-α)were detected by ELISA.The expression levels of PPAR-γ/NF-κB signaling pathway-related proteins were detected by immunohistochemistry and Western Blot.Results Compared with the normal group,the RVSP,mPAP,RVHI,RVIDd and RVAWT of the model group were significantly increased(P＜0.01).PAAT,PAAT/PET and TAPSE were significantly decreased(P＜0.01).The wall of pulmonary arterioles was significantly thickened,and the percentage of wall thickness of pulmonary arterioles to vascular diameter and the percentage of vascular wall area to total cross-sectional area of pulmonary arterioles were significantly increased(P＜0.01).The positive expression rate of α-SMA protein in pulmonary artery was significantly increased(P＜0.01).The levels of plasma IL-1β,IL-6 and TNF-α were significantly increased(P＜0.01).The positive expression rate of PPAR-γ protein in lung tissue was significantly decreased(P＜0.01),and the positive expression rate of NF-κB protein was significantly increased(P＜0.01).The protein expressions of PPAR-γ and IκB-α in lung tissue were significantly down-regulated(P＜0.01).The protein expression ratio of p-NF-κB/NF-κB was significantly increased(P＜0.01).Compared with the model group,RVSP,mPAP,RVHI,RVIDd and RVAWT in each administration group were significantly decreased(P＜0.05,P＜0.01),while PAAT,PAAT/PET and TAPSE were significantly increased(P＜0.05,P＜0.01).The thickness of the vascular wall was significantly reduced,and the percentage of the wall thickness of the pulmonary arterioles to the diameter of the blood vessels and the percentage of the vascular wall area to the total cross-sectional area of the small arteries were significantly reduced(P＜0.05,P＜0.01).The positive expression rate of α-SMA protein in pulmonary artery was significantly decreased(P＜0.05,P＜0.01).The plasma levels of IL-1β,IL-6 and TNF-α were significantly decreased(P＜0.05,P＜0.01).The positive expression rate of PPAR-γ protein in lung tissue was significantly increased(P＜0.05,P＜0.01),and the positive expression rate of NF-κB protein was significantly decreased(P＜0.05,P＜0.01).The protein expression of PPAR-γ in lung tissue was significantly up-regulated(P＜0.05,P＜0.01),and the protein expression ratio of p-NF-κB/NF-κB was significantly decreased(P＜0.01).The protein expression of IκB-α in the lung tissue of rats in the high-dose group of Feixin Decoction was significantly up-regulated(P＜0.01).Conclusion Feixin Decoction can improve pulmonary artery pressure,right ventricular dysfunction and pulmonary vascular remodeling in PAH rats induced by monocrotaline.The mechanism may be related to the regulation of PPAR-γ/NF-κB signaling pathway to inhibit inflammatory response.

OBJECTIVE: To systematically evaluate the efficacy difference between warming acupuncture and other acupuncture methods in the treatment of primary obesity. METHODS: A computer-based retrieval was conducted at PubMed, EMBASE, CENTRAL, CINAHL, Alt HealthWatch, CNKI, CBM, WANFANG database and VIP database. Retrieval time was from the establishment date of database to October 4, 2017. Randomized controlled trial (RCT) of warming acupuncture comparing with other acupuncture methods for the treatment of primary obesity were included. The relative risk () and weighted mean difference ( ) were used as combined effects for categorical variables and continuous variables, respectively. RESULTS: Totally 13 RCTs were included involving 878 patients. The Meta-analysis indicated compared with other acupuncture methods, warming acupuncture could more reduce weight (: -1.49 kg, 95% : -2.53 to -0.45, =0.005), improve the total effective rate (=1.16, 95% : 1.09 to 1.24, <0.000 01), reduce BMI (: -1.24 kg/m, 95% : -2.34 to -0.14, =0.03), reduce waist circumference (: -1.65 cm, 95% : -2.53 to -0.76, =0.02) and reduce hip circumference (: -2.86 cm, 95% : -4.37 to -1.35, =0.000 2), but had no significant influence on total cholesterol (: -0.05 mmol/L, 95% :-0.98 to 0.88, =0.91). CONCLUSION The warming acupuncture has better efficacy on primary obesity than other acupuncture methods, but less effects on lipid indicators.
Acupuncture Therapy ; Databases, Factual ; Humans ; Lipids ; Obesity ; therapy ; Randomized Controlled Trials as Topic

The incidence rate of nonalcoholic fatty liver disease (NAFLD) is increasing. Diet is considered one of the main driving forces regulating the composition of intestinal microbiota, and the intestine and the liver are closely linked through the portal vein, so changes in gut microbiota may affect liver function and promote inflammation, insulin resistance, and steatosis, thereby causing NAFLD. This article elaborates on the relationship between diet, gut microbiota, and the liver and the research advances in how this axis promotes the progression of NAFLD, as well as the change in potential mechanism due to intestinal dysbacteriosis and related treatment methods.