This experiment aimed to explore and research the process of preparing baicalein and wogonin through liquid fermentation with Bacillus natto. Active enzymes of produced by B. natto was used for the biological transformation of baclin and wogonoside, in order to increase the content of the haicalein and wogonin in the scutellaria. With the content of the baicalein and wogonin as evaluating indexes, the effects of carbon source, nitrogen source, the types and suitable concentration of inorganic salt, medium pH, granularities of medical materials, liquid volume in flask, shaking speed, liquid-to-solid ratio, fermentation time on the fermentation process were studied. The optimal process conditions for liquid fermentation of scutellaria were 1.0% of peptone, 0.05% of NaCl, pH at 6, the granularities of medical materials of the scutellaria screened through 40-mesh sifter, 33% of liquid, shaker incubator speed at 200 r x min(-1), liquid-to-solid ratio of 5:1, temperature at 37 degrees C, fermentation for 6 days, baclin's conversion rate at 97.6% and wogonoside's conversion rate at 97% in the scutellaria. According to the verification test, the process was stable and feasible, and could provide data reference for the industrial production.
Bacillus subtilis ; metabolism ; Biotransformation ; Fermentation ; Flavanones ; metabolism ; Flavonoids ; metabolism ; Glucosides ; metabolism ; Soy Foods ; microbiology

AIM: To discuss the protective effects and possible mechanisms of 17β-estradiol on human retinal pigment epithelial ( RPE) cells induced by high glucose. ·METHODS: RPE cells were cultured and divided into four groups according to randomized controlled method:blank control group:the cells were treated with 5. 5mmol/L routine glucose medium for processing; high glucose group: cells were treated with 100mmol/L glucose for 12h;17β-estradiol low concentration group: after treated with 10 μmol/L 17β-estradiol, cells were treated with 100mmol/L glucose for 12h; 17β-estradiol high concentration group: after treated with 100 μmol/L 17β-estradiol, cells were treated with 100mmol/L glucose for 12h. Cell viability were tested by MTT colorimetric detection. Cells apoptosis were detected by Hochest33258 staining. Intracellular reactive oxygen species( ROS) level were detected by H2 DCFDA staining. Expression of CAT, SOD and MDA were tested by colorimetric detection. · RESULTS: RPE cell activity decreased with the concentration of glucose increased; 17β-estradiol inhibited high glucose-induced cell viability decrease in RPE cells, decreased the apoptosis rate of RPE cells and intracellular ROS generation; besides, 17β-estradiol significantly increased the expression of CAT, SOD and decreased the expression of MDA in RPE cells. ·CONCLUSION: The 17β-estradiol effectively inhibited high glucose -induced RPE cells damage, which provide reliable experimental basis for the treatment of injuries in RPE cells.

Objective To retrospectively evalute the value and accuracy of adenosine stress and rest SPECT myocardial perfusion imaging. Methods A total of 1858 patients who were suspected or known for coronary artery disease (CAD) underwent 99Tcm-methoxyisobutylisonitrile ( MIBI ) myocardial perfusionSPECT with adenosine infusion using the standard 2-day protocol. Images were interpreted by two or more experienced nuclear medicine physicians . Coronary angiography was carried out in all patients within one month. Kappa test was used to analyze the correlation between the two imaging studies. Results By coronary angiography, there were 957 patients diagnosed of CAD (one-, two-, three-vessel disease: 506,256,195, respectively) and 901 normal. Stenosis was found in 1603 vessels, including left anterior descending coronary artery (LAD): 765, left circumflex coronary artery (LCX): 399 and right coronary artery (RCA): 439. By adenosine induced stress myocardial perfusion imaging, 876 patients were diagnosed of myocardial ischemia ( sensitivity: 876/957, 91.54% ) and 651 patients had negative findings ( specificity:651/901,72.25 % ). The positive and negative predictive values were 77.80% ( 876/1126 ) and 88.93% (651/732), respectively. The correlation coefficient between the two imaging studies was 0.641. The vessel-based sensitivity was 81.31% (622/765) for LAD, 56.64% (226/399) for LCX and 70.62% (310/439) for RCA, respectively. The sensitivity for detection of one-, two-, three-vessel stenosis was 87.55% (443/506), 94.92% (243/256) and 97.44% (190/195), respectively. The side-effects was mild and transient with an incidence rate of 84.12% ( 1563/1858), without major cardiac events. Conclusion Stress myocardial perfusion imaging induced by adenosine is reliable for the evaluation of myocardial blood supply in CAD patients.

Objective To predict the trend of hepatocellular carcinoma (HCC) mortality and investigate the features of its mortality including age, period, and birth cohort in males living in Haimen city of Jiangsu province, China. Methods Grey model (GM) was modeled using standardized mortality rate (SMR) of HCC from 1993 to 2006, and was applied to predicting SMR until 2012. Based on the mortality density (MD) for a four-year period, the goodness-of-fit of models and comparisons between models were evaluated so as to obtain the best one among these models including the effects of intercept, age-period-cohort (APC) , age-period (AP), age-cohort (AC),period-cohort(PC), and APC. Both APC full model and the best model were used to estimate effects of age, period, and cohort on HCC mortality. In addition, MD from 2005 to 2012 was predicted by the best model. Results Predictions based on GM (1,1 )showed that SMR was 48.578 per 100 000 population (relative error=-1.267% ) in 2007 year, which declined between 2008 and 2012. The lowest value was 45.578 per 100 000 people (in the 2012 year). The results of fitted models and comparisons between models showed that AP model was the best one (△G2=9.065,AIC=202.544). The curvatures of the effects of the three factors from APC model suggested that significances existed in changes of curvatures of 36.5-40.5 years old- (-0.368) and 64.5-68.5 years old-(-0.489) as well as in the change of 1956-1959 birth cohort (C21949.5. 1967.5=-0.492). The estimation of relative risks for AP model showed that the age effects were upward to 64.5-68.5 years old-, then downward; and that the period effects were found to be declined between 1993 and 2004. Predictions based on AP model suggested the decrease of HCC mortality. Conclusion The slightly decreasing trend of HCC mortality for males might be explained by age, period and a minor birth cohort effects in Haimen of China.

The different biological functions were studied in mouse bone marrow-derived endothelial progenitor cells isolated by differential time attachment to obtain the optimal adherent time in this study. Density gradient centrifugation-isolated bone marrow mononuclear cells were seeded on the fibronectin-coated dish. The 1-day cultured unattached cells were seeded on the second dish for 2 more days. Then unattached cells in the second dish were seeded on the third dish. The cells on 3 dishes were defined as 1-day adherent cells, 3-day adherent cells and 3-day unattached cells, respectively. After 20-day culture, the biological functions, such as the percentage of biomarkers, the ability of adhesion, and the ability of forming tubes in vitro were analyzed. The results showed that the percentages of positive CD34, FLK-1, and CD34/FLK-1 expressions in 1-day attached cells were significantly increased compared to those in the 3-day adherent or unattached cells (P < 0.01), which showed the strongest adhesion ability. The expression of eNOS in 1- or 3-day adherent cells was significantly higher than that in 3-day unattached cells (P < 0.01). The expression of VEGF in 3-day adherent cells was significantly higher than that in 1-day adherent cells or 3-day unattached cells (P < 0.01). These results suggest the biological functions of 1-day adherent cells are significantly stronger than that of 3-day adherent or unattached cells. VEGF expression in 3-day adherent cells is higher than that in 1-day adherent cells or 3-day unattached cells. The expression of eNOS in 1-day adherent cells or 3-day adherent cells is higher than that in 3-day unattached cells. The optimal adherent time to obtain mouse bone marrow-derived endothelial progenitor cells is 1-3 d.
Animals ; Bone Marrow Cells ; cytology ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Separation ; methods ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; Leukocytes, Mononuclear ; cytology ; Male ; Mice ; Mice, Inbred C57BL ; Nitric Oxide Synthase Type III ; metabolism ; Stem Cells ; cytology ; metabolism ; Time Factors ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo describe the clinical features, treatments and prognosis of very low birth weight infants (VLBWIs) requring mechanical ventilation, to assess the risk factors associated with the mortality of VLBWIs, and to evaluate the significance of the scoring system based on clinical risk index for babies (CRIB) and the score for neonatal acute physiology-perinatal extension II (SNAPPE-II) for predicting mortality risk for premature infants in China.

METHODSPerinatal data were collected from 127 VLBWIs requring mechanical ventilation who were admitted to the neonatal intensive care unit (NICU) from January 2010 to October 2011.

RESULTSThe enrolled infants had a mean gestational age of 31±2 weeks, a mean birth weight of 1290±170 g, a male/female ratio of 1.23∶1, and extremely low birth weight infant accounting for 6.3%. Of the 127 cases, 48.0% were administered with pulmonary surfactant (PS), and 49.6% received endotracheal intubation ventilation. The overall in-hospital mortality was 41.7%. Multivariate logistic regression revealed the following independent risk factors for mortality: low birth weight, multiple birth, cesarean section, and low PaO2/FiO2 ratio (OR = 1.611, 7.572, 4.062, and 0.133 respectively; P<0.05). SNAPPE-II and CRIB showed good performance in predicting prognosis, with areas under the ROC curve of 0.806 and 0.777 respectively.

CONCLUSIONSThe overall mortality rate of VLBWIs is still relatively high. The high-risk factors for VLBWI mortality include low birth weight, multiple birth, cesarean section, and low PaO2/FiO2 ratio. The neonatal illness severity scoring system (using SNAPPE-II and CRIB) can be used to quantify illness severity in premature infants.

Adult ; Female ; Hospital Mortality ; Humans ; Infant, Newborn ; Infant, Very Low Birth Weight ; Logistic Models ; Male ; Prospective Studies ; ROC Curve ; Respiration, Artificial ; Risk Factors

Flavonol synthase (FLS) is one of the key enzymes in flavonoids metabolic pathways. In this study, middle sequence was obtained from Carthamus tinctorius transcriptome sequencing results. Full-length cDNAs of FLS was cloned from petals of C. tinctorius to FLS by using RT-PCR and RACE technology. Its full-length cDNA was 1,201 bp, with an open reading frame of 1,101 bp and 336 encoded amino acids. The phylogenetic analysis showed that, FLS gene encoded amino acids in C. tinctorius were highly homologous with amino acids in congeneric Compositae species, especially Rudbeckia laciniata. The pBASTA-FLS plant expression vector was successfully built by the molecular biology method, which lays a foundation for further studying biology functions of the gene and biosynthesis mechanism of flavonoids.
Amino Acid Sequence ; Base Sequence ; Carthamus tinctorius ; classification ; enzymology ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; metabolism ; Molecular Sequence Data ; Open Reading Frames ; Oxidoreductases ; genetics ; metabolism ; Phylogeny ; Plant Proteins ; genetics ; metabolism

Objective:To investigate the quality regionalization and environmental impact factors of Stephaniae Tetrandrae Radix based on main active ingredients，and provide a reference for the determination of high-quality production areas and the dominant environmental factors affecting the content of Stephaniae Tetrandrae Radix. Method:Partial least squares regression analysis (PLS) and principal component analysis (PCA) were used to study the quality regionalization and environmental impact factors based on the main active ingredients of tetrandrine and fangchinoline, and investigate the environmental factors of the producing areas. Result:The content of fangchinoline was positively correlated with soil pH and annual average temperature，negatively correlated with latitude. The content of tetrandrine was positively correlated with soil pH，negatively correlated with annual rainfall and longitude. The total content was positively correlated with soil pH and annual average temperature，but negatively correlated with annual rainfall，latitude and longitude. Principal component analysis showed that the 50 production areas could be divided into four groups of quality formation. The groups with the highest scores were Shixing county in Guangdong，Shexian county in Anhui，Songxi county in Fujian，Nanxiong city in Guangdong and Xiangxiang city in Hunan，all of which were best areas for accumulation of the two main active ingredients. Conclusion:Soil pH，annual average temperature，annual rainfall，latitude and longitude are the main environmental factors affecting the main active ingredients of Stephaniae Tetrandrae Radix. The best areas for accumulation of tetrandrine and fangchinoline are Shixing county in Guangdong，Shexian county in Anhui，Songxi county in Fujian，Nanxiong city in Guangdong and Xiangxiang city in Hunan.