2.Application of Response Surface Methodology in Optimization of Precursors for Taxol Production by Fusarium mairei K178
Wen-Liang DAI ; Long CHENG ; Wen-Yi TAO ;
China Biotechnology 2006;0(11):-
The concentration of precursors for the production of taxol with Fusarium mairei K178 was optimized by response surface methodology.Firstly,Plackett-Burman design was undertaken to evaluate the effects of eight factors.With statistic regression analysis,the significant factors affecting taxol production were determined as follows: phenylalanine,sodium acetate and sodium benzoate.Box-Behnken design was used to optimize the three critical internal factors mentioned above,and the optimum concentration levels and the relationships among these factors was found out.By quadratic regression model equation with Design-Expert statistic methods,the optimal concentration of the variables were determined as: phenylalanine 2.8mg/L,sodium acetate 3.3g/L,sodium benzoate 31.8mg/L.Under such conditions,the taxol production was increased to 242.6?g/L,which was 15.6% higher than the maximum value in the single factor tests.The experiment values under the optimal conditions agreed with the predicted values,which indicated that the model was proper and effective.
3.Treatment of deep wound infection with fibrin glue amikacin complex
Zhijian MA ; Hongchang YANG ; Zhong CHEN ; Zhaoxiang WU ; Tao LI ; Gang CHENG ; Peng LIAO ; Yi OU ; Canzhang LI ; Tao HAN
Chinese Journal of Trauma 2009;25(6):554-557
Objective To discuss the efficiency of fibrin glue amikacin complex in controlling infection by observing the changes of leukocyte and neutrophilic granulocyte classifying counts after fibrin glue amikacin complex treated deep wound. Methods Clinical case-control study was used in the study. All patients were divided randomly into test group (100 patients) and control group (100 pa-tients), matched by wound location, wound size, time from injury to operation, combined injury and gen-eral antibiotics use to compare leukocyte and neutrophilic granulocyte classifying counts between both groups and observe possible toxic and side-effect in test group. Results Firstly, the test group and control group had the comparability in aspects of gender distribution, average age and injury mechanism (P >0.05). Secondly, there was statistical difference in classifying counts of leukocyte and neutrophilic granulocyte in the test group at different time points (P <0.05). The classifying counts of leukocyte and neutrophilic granalocyte peaked at 24 hours after operation, then decreased with treatment time and reached the lowest at 24 days after surgery or at day 1 before discharge. Thirdly, there existed statistical significance upon leukocyte counts in the test group and control group except for at day 1 after operation (P > 0.05), with lower counts in test group than control group. In aspect of neutrophil classifying counts, there was statistical significance (P < 0.05) at other time points in beth groups except for time points at days 1,2 and 12 (P >0.05). The test group had lower neutrophil classifying counts compared with con-trol group at different time points. Conclusion The fibrin glue amikacin complex has good clinical effort and high security, with no toxic or side effort in treatment of deep wound infection, and is worth clinical applicaiton.
4.Effect of docosahexaenoic acid on hepatic ischemia-reperfusion injury in rats
Tao ZHANG ; Guang YANG ; Huanhuan XIU ; Yi MA ; Donghui CHENG ; Wenqi HUANG
Chinese Journal of Anesthesiology 2015;35(9):1071-1074
Objective To evaluate the effect of docosahexaenoic acid (DHA) on hepatic ischemia-reperfusion (I/R) injury in rats.Methods Fifteen male Sprague-Dawley rats, aged 8-10 weeks, weighing 250-300 g, were randomly divided into 3 groups (n =5 each) using a random number table: sham operation group (group S), hepatic I/R group (group I/R) , and group DHA.Hepatic I/R was induced by clamping the hepatic pedicle supplying the left and middle lobes of the liver for 60 min, followed by 24 h reperfusion in anesthetized rats.DHA 4 mg/kg was injected intravenously at 30 min before ischemia and 10 min of reperfusion in group DHA.The equal volume of solvent was given instead in S and I/R groups.Blood samples were taken from the inferior vena cava at 24 h of reperfusion for determination of serum alanine aminotransferase (ALT), and aspartate aminotransferase (AST) activities, and resolvin D1 concentrations.The rats were then sacrificed, and the livers were removed for determination of myeloperoxidase (MPO) activity (by spectrophotometry), and interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) mRNA expression (by quantitative real-time reverse transcriptase polymerase chain reaction).The livers were cut into sections which were stained with haematoxylin and eosin, and examined under light microscope.Results Compared to group S, the serum ALT and AST activities, serum resolvin D1 concentrations, and MPO activity, and IL-6 and TNF-α mRNA expression in liver tissues were significantly increased in I/R and DHA groups (P<0.05).Compared to group Ⅰ/R, the serum resolvin 1D1 concentrations, and MPO activity and TNF-α mRNA expression in liver tissues were significantly decreased (P<0.05) , and no significant difference was found in the serum ALT and AST activities in group DHA (P>0.05).There was no significant difference in pathological changes of the liver between group DHA and group I/R.Conclusion DHA can attenuate inflammatory responses during hepatic I/R, but it is not sufficient to mitigate liver injury in rats.
5.Comparison of cerebral white matter fractional anisotropy and metabolite concentrations measured by 1 H-MRS in healthy adults
Sainan CHENG ; Qiang LIU ; Yubo Lü ; Yuchao LI ; Tao GONG ; Yi ZHANG
Journal of Practical Radiology 2014;(10):1614-1617
Objective To investigate the relationship between cerebral white matter fractional anisotropy(FA)and metabolite con-centrations measured by 1 H magnetic resonance spectroscopy(1 H-MRS)in healthy adults.Methods Diffusion tensor imaging(DTI) and 1 H-MRS were acquired from thirty-one healthy adults at 3.0 T MR system.The concentrations of N-acetylaspartate(NAA), choline(Cho),creatine(Cr),the ratios of NAA/Cr and FA values of DTI were measured in 353 voxels,covering the bilateral medium corona radiata.According to the age,all voxels were divided into two groups:the senior group (mean age=29 years),and the senior group(mean age=5 6 years).The correlations between FA values and age were analyzed using Pearson’s correlation coefficients.The correlations between FA values and metabolite concentrations were estimated using partial correlations test controlling for the age re-lated bias.Results Pearson’s correlation analysis revealed a negative correlation between FA values and age in 353 voxels(r=-0.146,P<0.05),senior group(r=-0.204,P<0.05)and senior group(r=-0.162,P<0.05).Only the correlation between NAA and FA values were significant(r=0.339,P<0.05;r=0.213,P<0.05;r=0.430,P<0.05).There were positive relation-ships between FA values and NAA/Cr among 353 voxels (r=0.166,P<0.05)and senior group(r=0.305,P<0.05).Conclusion There are correlations between cerebral white matter FA values and metabolite concentrations measured by 1 H-MRS .Combining 1 H-MRS with DTI can reveal the relationship between microscopic white matter integrity and metabolic characteristics of the white matter.
6.Study on hydrolysis of baicalin into baicalein by immobilized beta-glucosidase in a two-phase system.
Yi-Shun YANG ; Tao CHENG ; Jun YANG ; Tong ZHANG ; Zhen-Zhen CAI
China Journal of Chinese Materia Medica 2014;39(4):669-673
The optimum conditions of baicalin hydrolysis into baicalein by immobilized beta-glucosidase in a two-phase system was studied and the yield was observed. A two-phase system comprising of sodium acetate buffer and chloroform was determined by comparing the solubleness of baicalein in different solvents and partition coefficient of baicalein in related aqueous-organic two-phase system. beta-Glucosidase was immobilized by the crosslinking-embedding method using sodium alginate as the carrier The optimum reaction temperature, pH value, Michaelis constant, the thermal stability and pH stability were assayed. By comparing the yield of baicalin hydrolysis into baicalein by immobilized beta-glucosidase in two-phase system, the optimum reaction conditions were determined-the optimum reaction temperature, pH value and time were 50 degrees C, 5.0 and 10 h, respectively. The yield of baicalein was 85.28%. Compare with one-phase system, two-phase system had an advantage in reaction rate and yield.
Biocatalysis
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Drugs, Chinese Herbal
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chemistry
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Enzyme Stability
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Enzymes, Immobilized
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chemistry
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Flavanones
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chemistry
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Flavonoids
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chemistry
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Hydrolysis
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beta-Glucosidase
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chemistry
7.Clinical characteristics and outcome of novel coronavirus pneumonia patients with different body mass index
Min WANG ; Fang YANG ; Xinxin ZHU ; Chuanfang CHENG ; Qi LI ; Tao ZHAO ; Yi ZOU ; Yimin YAN
Chinese Journal of Endocrinology and Metabolism 2021;37(1):17-22
Objective:To investigate the clinical characteristics and outcome of novel coronavirus pneumonia (COVID-19)patients with different body mass index (BMI), and to provide the basis for disease assessment and prognosis.Methods:The clinical data of 541 patients with COVID-19 diagnosed in Xiaogan Hospital Affiliated to Wuhan University of Science and Technology from January 16 to March 28, 2020 were collected. The patients were divided into normal weight group, overweight group, and obesity group according to BMI. The clinical characteristics and outcomes of the three groups were compared. The correlation between BMI and clinical classification was analyzed by ordinal logistic regression.Results:There were 288 cases (53.23%) in normal weight group, 193 cases (35.67%) in overweight group, and 60 cases (11.09%) in obesity group. Compared with normal weight group, overweight and obesity groups displayed higher proportion of hypertension, with increased levels of white blood cells, neutrophils, C reactive protein, aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, and triglyceride in laboratory test results, and higher proportion of severe and critical illness ( P<0.05 or P<0.01). After adjusting for age, gender, and underlying diseases, regression analysis showed that higher BMI predicted more severe clinical classification ( OR=1.079, 95% CI 1.010-1.154). Conclusion:In COVID-19 patients, overweight and obese patients are more likely to develop into severe and critical illness, suggesting that obesity may be an important risk factor affecting the clinical outcome of COVID-19.
8.The Research of Enzymology Characterization about Arginine Deiminase from Enterococcus faecalis
Cheng-Fu LI ; Kai LI ; Jia-You LI ; Qing-Cai JIAO ; Qian LIU ; Li-Tao YI ;
Microbiology 1992;0(06):-
Arginine Deiminase(ADI) was purified to homogeneity using ammonium sulfate precipitation,Q-Sepharose Fast Flow anion exchange chromatography and SephadexG-75 gel filtration chromatography. This purification protocol resulted in a 34.5-fold purification of ADI with 31.4% final yield. A molecular weight of about 190 kD determined by native gradient polyacrylamide gel electrophoresis. The enzyme has only one kind of 46 kD subunit determined by SDS-PAGE. Combining the results from the two kinds of electrophoresis,the authors deduce that the enzyme may be a tetramer. The optimum pH and temperature for lipolytic activity of ADI was pH 6.5 and 50℃,respectively. It was extremely stable at 45℃ and retained 97.9% of its original activity for 30 min. The stability declined rapidly as soon as the temperature rose over 50℃. ADI was highly stable in the pH range from pH 5-8. ADI acted on L-arginine but not on D-arginine. ADI catabolism was dependent on metal ions. At their adequate concentration,Mn2+,Mg2+ and Co2+ were the effective promoter,while superfluous Zn2+and Co2+ inhibited ADI activity. L-citrulline did not act on ADI,but L-ornithine inhibited ADI activity. The degradation of L-arginine with ADI catalysis was according to simple Michaelis-Menten equation. The Michaelis constant was 3.2686 mmol/L and the maxi-mum velocity was 2.44 ?mol/min.
9.Serum Uric Acid and Prehypertension
Tao CHEN ; Wei LI ; Bo HU ; Yang WANG ; Xiao-Ru CHENG ; Xuan JIA ; Yi SUN ;
Chinese Journal of Hypertension 2006;0(08):-
Background Previous experimental and clinical studies have proved that elevated serum uric acid increased risk for developing hypertension.Whereas,there are a paucity of information on the relationship between serum uric acid and prehypertension.Objective The purpose of this research is to evaluate the association between the serum uric acid and prehypertension.Method A cohort of seven thousand eight hundred thirty-nine subjects without hypertension and other cardiovascular diseases were recruited from a cross sectional study in urban and rural place in 9 provinces during 2005-2006.Based on serum uric acid(324 ?mol/L for overall population,366 ?mol/L for male,285 ?mol/L for female),people were categorized into quartiles.The odds ratio for prehypertension was calculat ed with the lowest quartile as the reference.Results The prevalence of prehypertension increased with increasing uric acid in total population(P324 ?mol/L)to lowest quartile 1(
10.Study on a Novel Hepatitis B Virus Multi-epitope DNA Vaccine with siRNA,hIL-12 Coexpressing
Jing-Xian HUANG ; Yi-Cheng CAO ; Zheng-Ping DU ; Chang-Li TAO ; Hua-Qiang YANG ;
China Biotechnology 2006;0(08):-
Objective:To construct a DNA vaccine co-expressing the HBV compound multi-epitope antigen gene, the hIL-12 and the anti-HBV siRNA genes, and to express this DNA vaccine in HepG2 cells. Methods:The HBV multi-epitope antigen gene was designed and synthesized before it was fused with enhanced green fluorescent protein(EGFP) gene, and cloned into the multi-clone site(MCS) of the eukaryotic expression vector pVAX1. The expressinig units of hIL-12 and siRNA were cloned into the BspH I and Mlu I site of pVAX1 respectively. Then the recombinant plasmid pVAX1-siHBV-HB-EGFP-hIL12 was transiently transfected HepG2 cells. The expression of HBV compound multi-epitope gene was observed through EGFP report gene. The expression of hIL-12 was analyzed by ELISA and the effects of anti-HBV siRNA was confirmed with rtPCR . Results: The analysis of enzyme digestion and sequencing both demonstrated that the trible-expressing HBV DNA vaccine has been constructed successfully. The green fluorescent image was detected in the transfected cells which could confirm the expression of the multi-epitope antigen gene. The amount of hIL-12 secretion was 1289pg/ml in supernatant at 48h after transfection and 1712pg/ml at 72h after transfection. The mRNA amount of HBV S gene, which was the siRNA target, had been obviously knockdown. Conclusion: The DNA vaccine co-expressing the HBV compound multi-epitope antigen gene, the hIL-12 and the siRNA genes was constructed and transiently expressed in HepG2 cells, and siRNA had shown us a good anti-HBV effect. It laid a foundation of further study on anti-HBV effect of the new DNA vaccine.