Using chemically defined medium as the control, mechanism of corn steep liquor (CSL) in complex medium during glycerol production by Candida glycerinogenes was studied.The results showed that there were three key factors in CSL that had some great influences on glycerol fermentation of C.glycerinogenes, including phosphorus, nitrogen, and trace elements.The maximum glycerol yield of 53.44% was achieved at an optimal phosphorus concentration of 121.75mg/L, where the CSL concentration was 14g/L.Phosphorus in CSL could control the distribution of carbon metabolism flux between EMP pathway and HMP pathway.With the increase in CSL concentrations, superfluous phosphorus could restrain HMP pathway and activate EMP pathway, thus resulting in remarkable changes in various fermentation parameters of complex medium.Nitrogen in CSL could play a cooperative role in the regulative function of phosphorus.However, it was not a suitable nitrogen source for C.glycerinogenes.Trace elements in CSL could markedly improve the glucose consumption rate, accelerate the cell growth, and enhance the glycerol yield.

In the reductive branch, glycerol is first dehydrated to 3-hydroxypropionaldehyde that is then reduced to 1,3-PD under the consumption of reducing power (NADH). If over-expression of the gene dhaB encoding glycerol dehydrase is achieved,the reducing power will be scarce and 3-hydroxypropionaldehyde will be accumulated,which is disadvantage to produce 1,3-propanediol.The structure gene yqhD from E.coli encoding 1,3-propanediol oxidoreductase isoenzyme[under the consumption of reducing power (NADPH)]and the gene dhaB encoding glycerol dehydrase from Klebsiella pneumoniae was amplified using PCR method.The two gene were transferred into expression vector pEtac to construct a novel recombinant Klebsiella pneumoniae (pEtac-dhaB-tac-yqhD).Over-expression of yqhD and dhaB in Klebsiella pneumoniae was achieved with pEtac-dhaB-tac-yqhD.The fermentation result on aerobic conversion showed the increase of 20% of 1,3-propanediol yield by Klebsiella pneumoniae(pEtac-dhaB-tac-yqhD) was obtained compared with Klebsiella pneumoniae.The main by-products,acetic acid and butanediol decreased estrogen receptors 30% obviously.

Aged, 80 and over ; Echinocandins ; adverse effects ; therapeutic use ; Humans ; Lipopeptides ; Liver ; drug effects ; Liver Function Tests ; Mycoses ; drug therapy ; Retrospective Studies

To study the bioactive polypeptides included in Bufo skin and its secretions the plasmid skin cDNA library of adult Japanese toad Bufo japonicus formosus was prepared. The pSD64TR has been used as the vector and the cloning sites are Xho I and EcoR I. To screen cDNAs encoding bioactive components, the plasmid cDNA library was transformed into E. coli DH5 competent cells, and positive colonies were screened by colony PCR (polymerase chain reaction). The suspension of a single colony in LB medium was used as the template, SP6 (the upstream primer of the plasmid cDNA library) and a primer with Xho I site and polyT were used as the primers. As the result, 465 positive colonies out of 1 344 were obtained and their plasmid were collected and sequenced. By homologous analysis, it was found that one of the cDNAs encoding a peptide with high homolog with transgelin-2, which was registered in GenBank (accession number: JX197456), and it was indicated as a partial cDNA sequence with a deletion at the 5' end. The transcript is 997 bp consisting of 31 bp 5', 618 bp 3' untranslated region (UTR) and an open reading frame (ORF) of 348 bp encoding a polypeptide of 115 amino acids. In the putative protein product, there is a calponin homology domain, two cysteine residues for a disulfide bond and three a-helix domains, and five potential phosphorylation sites. The homologous analysis indicates 90% similarity with Xenopus (Silurana) tropicalis and 89% with Xenopus laevis, and 71%-85% with other species.

The dhaD gene encoding glycerol dehydrogenase(GDH) from Klebsiella sp.was amplified,and was inserted into expression vector pET-28a(+),the plasmid pET-28a-dhaD was constructed and was transformed into Escherichia coli BL21(DE3).SDS-PAGE showed that the gene dhaD was expressed successfully in recombinant E.coli BL21.Then GDH was purified by Ni-NTA affinity chromatography,the results showed a single band about 39kDa on SDS-PAGE gel,and the specified activity was about 156U/mg.The special activity of GDH is 4.6-fold higher than that of unpurified and the activity recovery is 67.4%.The optimum reaction pH was 11.0,and the GDH activity have little changed when incubated in the buffer of pH7.0～11.0.The optimum reactive temperature was 30℃,and the GDH was more stable on the temperature of 25℃～45℃.The Km value was 0.54mmol/L and Vmax was 0.49 ?mol/ml?min in the glycerol.

Objective To study the effects of health management on cardiovascular events in the elderly patients with hypertension. Methods A total of 182 elderly patients with hypertension were randomly assigned to the control group (n = 61 ), pharmacologic therapy group (n = 61 ) or health management group ( n = 60). Serum biomarkers, brachial-ankle pulse wave velocity ( baPWV ), and blood pressure were tested at baseline and after intervention. Results There were no differences between the 2 groups in clinical characteristics at baseline. The average following-up period was (21 ± 7 ) months. The improvement of systolic blood pressure ( t = 3.915, P = 0. 000 ), pulse pressure ( t = 3. 966, P = 0. 000), and baPWV ( t = 3. 093, P = 0. 002) in the health management group was more significant than the control group;the systolic blood pressure ( t = 2. 008, P= 0. 046 ) was bitterly improved than the pharmacologic therapy group. The accumulative survival rate of the health management group (96. 7% ) was higher than the control group (83.6%; x2 =5. 921 ,P =0. 015) ,similar to the pharmacologic therapy group (93.3%; x2 =2. 821,P=0.091 ). Decreased systolic blood pressure, diastolic blood pressure, pulse pressure and baPWV were protective factors. After adjusted by age and gender,the improvement of systolic blood pressure was found to be an independent protective factors ( RR = 0. 75, P ＜ 0. 05 ). Conclusion Health management in elderly patients with hypertension could more significantly reduce the risk of cardiovascular diseases.

Objective To study the effects of health management on blood pressure and lifestyle of hypertensive outpatients.Methods A total of 319 hypertensive outpatients were randomly assigned to the control group (n =160) or the health management group (n =159).Pharmacologic therapy was given to the control group,while in the health management group,intensive health management combined with pharmacologic therapy was conducted.Blood pressure,height,body weight (BW),total cholesterol (TC),and fasting blood glucose (FBG) of the participants were measured and compared at baseline and 12 months.Results There were no significant differences of clinical characteristics between two groups at baseline.After 12 months' intervention,more significant decrease of systolic blood pressure and diastolic blood pressure was found in the health management group (t values were 2.701 and 2.306,respectively;both P ＜ 0.05).There were statistically significant differences of body mass index (BMI) and serum TC levels between the control and the health management group (t values were 2.111 and 2.227,respectively ;P ＜ 0.05).After the intervention,two groups showed no significant difference in current cigarette smoking (x2=2.787,P ＞ 0.05).The participants in the health management group showed improved physical exercises,diet,and adherence to treatment at the end of the observation (x2 values were 59.459,52.018,6.321 and 5.392,respectively; all P ＜ 0.05).Conclusion Compared with pharmacologic therapy,health management combined with pharmacologic therapy could significantly improve clinical parameters and life style of hypertensive patients.

ObjectiveTo study the effects of health management on elderly patients with metabolic syndrome and lower extremity arterial disease.Methods A total of 118 metabolic syndrome( MS ) patients with ankle-brachial index (ABI) ＜ 0.9 and no intermittent claudication were randomly assigned to the control group( n =60,receiving pharmacologic therapy ) or health management group ( n =58,receiving intensive health management + pharmacologic therapy).Blood pressure,ABI,total cholesterol (TC),and fasting blood glucose(FBS) were tested before and 12 after the intervention.Results Clinical data were comparable between the two groups at baseline.After 12-months' intervention,systolic blood pressure(SBP) and FBS in both groups were significantly decreased,especially in the health management group[SBP:(141 ± 10)mm Hg(1 mm Hg=0.133 kPa) vs(146±11) mm Hg,t =2.581,P=0.011; FBS:(5.32 ±1.33) mmol/L vs ( 5.92 ± 1.61 ) mmol/L,t =2.231,P =0.028].TC ( 5.51 ± 0.51 ) mmol/L vs ( 5.11 ± 0.49 ) mmol/L,(t=4.307,P=0.000) and ABI(0.77 ±0.17 vs 0.84 ±0.19,t=2.091,P=0.039) of the health management group were significantly improved at 12 months.Patients in the health management group also showed higher SF-36 scores than those in the control group( physical functioning:72.2 ± 12.4 vs 65.2 ±20.1,t =2.268,P =0.025 ; emotion:73.9 ± 18.0 vs 65.6 ± 21.1,t =2.295,P =0.023 ; mental health:63.9 ± 13.3vs 58.3 ± 12.5,t =2.358,P =0.020 ).Conclusion Compared with medication,health management and pharmacologic therapy could significantly improve clinical parameters and quality of life of elderly MS patients combined with peripheral arterial disease.

The hom gene encoding for homoserine dehydrogenase was amplified from the genomic DNA of Corynebacterium glutamicum ATCC 13032.After the kanamycin-resistant gene(Km)cassette from plasmid pET28a was inserted into the center of hom,the hom::Km cassette was then electroporated into the competent cell of C.glutamicum ATCC 13032.And kanamycin-resistant clones were obtained.PCR was performed to confirm whether the Km gene was integrated into the hom gene of these clones and the recombinant strains of hom-disrupted were screened out.Fermentation results showed that the lysine yield of the hom-disrupted strain C.g-hom::Km-8 reached 4.7 g/L,which was 6.7 times that of C.glutamicum ATCC 13032.

In order to isolate genes related with the osmoadaptation and glycerol metabolism of Candida glycerinogenes, a transformation system based on the dominant selectable marker Zeocin and restriction enzyme-mediated integration (REMI) was established. Effects of seven restriction enzymes on transformation efficiency of C.glycerinogenes were tested. Transformation conditions were optimized in the presence of Hind III. Under the optimal conditions of OD_ 600 ≈1.3, voltage of 1.5 kV, 2.0?10~9 competent cells/mL, 100 units of Hind III added, the transformation efficiency was up to 129 trnaformants/?g DNA. 58% of transformants were stable on nonselective medium. These results suggest that REMI technique would be beneficial to the genetic transformation of C.glycerinogenes.