Batch fermentation of lycopene by recombinant E. c ol i ZYL-2 at various temperatures ranging between 23℃～37℃ were studied in 7L fermentor. At 33℃, cell specific growth rate in earlier stage of culture was higher, and the time obtained maximum cell dry weight was shorter at other tempe rature. while after 9 h, lycopene specific production rate was higher at 28℃. Based on these results, a two-stage temperature control strategy was developed in which 33℃ was used for fermentation for the first 9 h and the temperaturew as switched to 28℃ after 9h. Using this temperature-shift strategy, the maxim al lycopene content and productivity reached 605.25?g L-1 and 28.82?g L-1h-1. The lycopene fermentation level obtained by the strategy was higher than those in single temperature-control experiments.

Animals ; Arrhythmias, Cardiac ; metabolism ; prevention & control ; Female ; Hyperthermia, Induced ; methods ; Ischemic Preconditioning ; methods ; Male ; Myocardial Reperfusion Injury ; metabolism ; prevention & control ; Rats ; Rats, Sprague-Dawley ; Stress, Physiological ; physiology ; Superoxide Dismutase ; blood

Acute Disease ; Animals ; Female ; Hypoxia ; physiopathology ; Male ; Microcirculation ; physiology ; Rats ; Splanchnic Circulation ; physiology ; Superoxide Dismutase ; blood

Clinical features and thyroid-stimulating antibody(TSAb)in 32 newly diagnosed patients with Graves' ophthalmopathy(GO)were compared with those in 27 Graves' disease(GD)patients without GO(as control group).All of the patients with GO received intravenous glucocorticoids.The level of serum TSAb in patients with GO was significantly higher than that in patients without GO.TSAb was also associated with the prognosis.It suggests that TSAb seems to be the most active component among the TSH receptor antibodies related to ophthalmopathy and may act as a predictive parameter.

Objective The presence of alloreactive memory T cells in recipient is a critical handicap to achieve transplantation tolerance.To make a mouse model which mimics the present transplant patient is important for research at this subject.Thus,we developed a novel re-transplant model and compared the alloresponse in this model with that in the conventional memory T cellstransfer model (transfer control).Methods The re-transplant model was established via microsurgery and vessel cannula techniques,and the experiment was composed of three groups:the re- transplant group,memory T cell-transfer group (transfer control) and the conventional blank group (blank control).The research indexes included survival time of donor heart,rejection score of allograft,and detection of proliferation and differentiation of the alloreactive memory/effector T cells by by flow cytometry (FCM) and in vitro mixed lymphocyte reaction (MLR).Results The median survival time of allograft in re-transplant recipients was significantly shortened compared to that of transfer control,but there was no significant difference in rejection score of graft between them (the score in retransplant group was the most intense of the three groups). Moreover, proliferation and differentiation of the alloreactive effector T cells were more intensive in re- transplant recipients than in the transfer control,which was confirmed by in vitro MLR and by FCM of the splenocytes for detecting CD44highCD62L-memory/effector phenotype cells.Conclusion The recall alloresponse in retransplantation is more intensive than that in memory-transfer setting and this re-transplant model is more close to the clinic situation than the memory-transfer model in rodents.

Taxol is one of the most potent anti-cancer agents, which is extracted from the plants of Taxus species. Isopentenyl diphosphate isomerase (IPI) catalyzes the reversible transformation between IPP and DMAPP, both of which are the general 5-carbon precursors for taxol biosynthesis. In the present study, a new gene encoding IPI was cloned from Taxus media (namely TmIPI with the GenBank Accession Number KP970677) for the first time. The full-length cDNA of TmIPI was 1 232 bps encoding a polypeptide with 233 amino acids, in which the conserved domain Nudix was found. Bioinformatic analysis indicated that the sequence of TmIPI was highly similar to those of other plant IPI proteins, and the phylogenetic analysis showed that there were two clades of plant IPI proteins, including IPIs of angiosperm plants and IPIs of gymnosperm plants. TmIPI belonged to the clade of gymnosperm plant IPIs, and this was consistent with the fact that Taxus media is a plant species of gymnosperm. Southern blotting analysis demonstrated that there was a gene family of IPI in Taxus media. Finally, functional verification was applied to identify the function of TmIPI. The results showed that biosynthesis of β-carotenoid was enhanced by overexpressing TmIPI in the engineered E. coli strain, and this suggested that TmIPI might be a key gene involved in isoprenoid/terpenoid biosynthesis.
Amino Acid Sequence ; Carbon-Carbon Double Bond Isomerases ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; Paclitaxel ; biosynthesis ; Phylogeny ; Plant Proteins ; genetics ; Taxus ; enzymology ; genetics

Hyoscyamine 6 beta-hydroxylase (H6H) is the last rate-limiting enzyme directly catalyzing the formation of scopolamine in tropane alkaloids (TAs) biosynthesis pathway. It is the primary target gene in the genetic modification of TAs metabolic pathway. Full-length cDNA and gDNA sequences of a novel H6H gene were cloned from Datura arborea (DaH6H, GenBank accession numbers for cDNA and gDNA are KR006981 and KR006983, respectively). Nucleotide sequence analysis reveals an open reading frame of 1375 bp encoding 347 amino acids in the cDNA of DaH6H, while the gDNA of DaH6H contains four exons and three introns, with the highest similarity to the gDNA of H6H from D. stramonium. DaH6H also exhibited the most identity of 90.5% with DsH6H in amino acids and harbored conserved 2-oxoglutarate binding motif and two iron binding motifs. The expression level of DaH6H was highest in the mature leaf, followed by the secondary root, and with no expression in the primary root based on qPCR analysis. Its expression was inhibited by MeJA. DaH6H was expressed in E. coli and a 39 kD recombinant protein was detected in SDS-PAGE. Comparison of the contents of scopolamine and hyoscyamine in various TAs-producing plants revealed that D. arborea was one of the rare scopolamine predominant plants. Cloning of DaH6H gene will allow more research in the molecular regulatory mechanism of TAs biosynthesis in distinct plants and provide a new candidate gene for scopolamine metabolic engineering.
Cloning, Molecular ; DNA, Complementary ; Datura ; enzymology ; genetics ; Escherichia coli ; Hyoscyamine ; chemistry ; Mixed Function Oxygenases ; genetics ; Plant Leaves ; enzymology ; Plant Roots ; enzymology ; Recombinant Proteins ; genetics ; Scopolamine Hydrobromide ; chemistry

This research was a part of the investigation of traditional Chinese medicine resources survey in Markam. The medicinal plants in natural reserve were studied for the first in this paper. There were 300 species in 202 genera of 54 families, among them there were 7 species of ferns in 5 genera of 5 families, 6 species of gymnosperms in 4 genera of 3 families, and 287 species of angiosperms in 194 genera of 61 families. There were 166 species Tibetan medicinal plants in 102 genera of 47 families. Quantitative analysis was carried out in 6 aspects of family and genus composition, medicinal parts, drug properties, flavour of a drug, Tibetan medicine, toxicity and new plants. The concrete suggestions of protection and exploitation were put forward, which provided scientific basis for the sustainable utilization of medicinal plants in this area.
Biodiversity ; Conservation of Natural Resources ; Medicine, Tibetan Traditional ; Plants, Medicinal ; Tibet

Objective To analyze personality characteristics and psychological situation of children suffered from nephrotic syndrome of school age,and to provide clinical evidence for the children patients to be interfered fur- ther by psychological act.Methods EPQ,SAS and SDS were used to survey the personality and psychology of 30 cases of children suffered from nephrotic syndrome of school age and 28 cases of children who were in normal school age as statistic calculation comparison.Results The score of N,P,L inventory for the children suffered from nephrotie syndrome of school age was higher,and the score of E inventory was lower,which all had remarkably(P

Artemisnin is a novel sesquiterpene lactone with an internal peroxide bridge structure, which is extracted from traditional Chinese herb Artemisia annua L. (Qinghao). Recommended by World Health Organization, artemisinin is the first-line drug in the treatment of encephalic and chloroquine-resistant malaria. In the present study, transgenic A. annua plants were developed by overexpressing the key enzymes involved in the biosynthetic pathway of artemisinin. Based on Agrobacterium-mediated transformation methods, transgenic plants of A. annua with overexpression of both HDR and ADS were obtained through hygromycin screening. The genomic PCR analysis confirmed six transgenic lines in which both HDR and ADS were integrated into genome. The gene expression analysis given by real-time quantitative PCR showed that all the transgenic lines had higher expression levels of HDR and ADS than the non-transgenic control (except ah3 in which the expression level of ADS showed no significant difference compared with control); and the HPLC analysis of artemisinin demonstrated that transgenic A. annua plants produced artemisinin at significantly higher level than non-transgenic plants. Especially, the highest content of artemisinin was found in transgenic line ah70, in which the artemisinin content was 3.48 times compared with that in non-transgenic lines. In summary, overexpression of HDR and ADS facilitated artemisinin biosynthesis and this method could be applied to develop transgenic plants of A. annua with higher yield of artemisinin.
Artemisia annua ; genetics ; metabolism ; Artemisinins ; metabolism ; Biosynthetic Pathways ; Drugs, Chinese Herbal ; Mixed Function Oxygenases ; genetics ; Oxidoreductases ; genetics ; Plant Proteins ; genetics ; Plants, Genetically Modified ; genetics ; metabolism ; Plants, Medicinal ; genetics ; metabolism