1.Improvement of Ethanol Fermentation by Expression of Schizosaccharomyces pombe mel Gene and Disruption of GPD1 in Saccharomyces cerevisiae
Microbiology 1992;0(06):-
Based on homologous recombination,?-galactosidase gene mel from S.pombe was integrated to the chromosomal DNA of industrial S.cerevisiae and the key enzyme gene GPD1 of the glycerol anabolic pathway was disrupted contemporaneously,and recombinants were screened through increasing G418 concentration.It was shown that melibiose utility capability of recombinant S.cerevisiae MG1 was increased evidently,glycerol productivity was decreased,and the mel gene was expressed stabilized in the host cell.No impact on the cell character was appeared after introduced extrinsic gene,but the cells were flocculate when growing.When compared with parent industrial S. cerevisiae Y to ferment with corn powder and wheat starch as raw substrates,ethanol productivity was increased,glycerol productivity was decreased,and the melibiose in the broth was exhausted completely detected by HPLC analysis.
2.Effects of overexpression of NADH kinase gene on ethanol fermentation by Saccharomyces cerevisiae.
Han WANG ; Liang ZHANG ; Guiyang SHI
Chinese Journal of Biotechnology 2014;30(9):1381-1389
Glycerol is the main byproduct in ethanol production by Saccharomyces cerevisiae. In order to improve ethanol yield and the substrate conversion, a cassette about 4.5 kb for gene homologous recombination, gpd2Δ::PGK1(PT)-POS5-HyBR, was constructed and transformed into the haploid strain S. cerevisiae S1 (MATa) to replace the GPD2 gene by POS5 gene. The NADH kinase gene POS5 was successfully over expressed in the recombinant strain S. cerevisiae S3. Comparing with the parent strain, the recombinant strain S. cerevisiae S3 exhibited an 8% increase in ethanol production and a 33.64% decrease in glycerol production in the conical flask fermentation with an initiatory glucose concentration of 150 g/L. Overexpression of NADH kinase gene seems effective in reducing glycerol production and increasing ethanol yield.
Ethanol
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chemistry
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Fermentation
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Glycerol
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chemistry
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Industrial Microbiology
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Mitochondrial Proteins
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genetics
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metabolism
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Phosphotransferases (Alcohol Group Acceptor)
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genetics
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metabolism
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Saccharomyces cerevisiae
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genetics
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metabolism
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Saccharomyces cerevisiae Proteins
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genetics
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metabolism
3.Recent research on tofacitinib in the treatment of pediatric rheumatic diseases.
Shi-Hai ZHOU ; Ya-Qun XIONG ; Ya CHEN
Chinese Journal of Contemporary Pediatrics 2022;24(4):447-453
Tofacitinib is a Janus kinase inhibitor and can block the Janus kinase-signal transducer and activator of transcription signal transduction pathway and reduce the production and release of a variety of cytokines. It has great potential in the treatment of various rheumatic diseases with a rapid onset of action and can reduce corticosteroid dependence and related adverse events. The therapeutic effect of tofacitinib in adult patients has been confirmed, and it has been increasingly used in pediatric patients in recent years. This article reviews the clinical application of tofacitinib in the treatment of pediatric autoimmune diseases.
Adult
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Child
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Humans
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Janus Kinases/metabolism*
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Piperidines/therapeutic use*
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Protein Kinase Inhibitors/therapeutic use*
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Pyrimidines/therapeutic use*
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Rheumatic Diseases/drug therapy*
4.Transformation of phosphotransferase system in Escherichia coli.
Mengrong XIAO ; Liang ZHANG ; Shuangping LIU ; Guiyang SHI
Chinese Journal of Biotechnology 2014;30(10):1561-1572
We constructed several recombinant Escherichia coli strains to transform phosphoenolpyruvate: carbohydrate phosphotransferase system (PTS system) and compared the characteristics of growth and metabolism of the mutants. We knocked-out the key genes ptsI and ptsG in PTS system by using Red homologous recombination in E. coli and meanwhile we also knocked-in the glucose facilitator gene glf from Zymomonas mobilis in the E. coli chromosome. Recombinant E. coli strains were constructed and the effects of cell growth, glucose consumption and acetic acid accumulation were also evaluated in all recombinant strains. The deletion of gene ptsG and ptsI inactivated some PTS system functions and inhibited the growth ability of the cell. Expressing the gene glf can help recombinant E. coli strains re-absorb the glucose through Glf-Glk (glucose facilitator-glucokinase) pathway as it can use ATP to phosphorylate glucose and transport into cell. This pathway can improve the availability of glucose and also reduce the accumulation of acetic acid; it can also broaden the carbon flux in the metabolism pathway.
Biological Transport
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Escherichia coli
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enzymology
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genetics
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Gene Deletion
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Gene Knock-In Techniques
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Gene Knockout Techniques
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Glucose
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metabolism
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Phosphoenolpyruvate Sugar Phosphotransferase System
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genetics
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Zymomonas
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genetics
5.Determining postmortem interval by accumulated temperature method.
Journal of Forensic Medicine 2006;22(6):438-458
The experience of bug's growing and accumulated temperatures were important ways for determination of postmortem interval in forensic science. Here we used reverse accumulated temperature methods to estimate postmortem interval and made accordant result with their true time.
Adult
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Animals
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Cadaver
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Cause of Death
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Entomology
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Female
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Forensic Pathology/methods*
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Humans
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Larva/growth & development*
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Life Cycle Stages
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Muscidae/physiology*
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Postmortem Changes
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Temperature
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Time Factors
6.Upregulated heme oxygenase-1 expression of mouse mesenchymal stem cells resists to chemotherapy-induced bone marrow suppression.
Shuya CHEN ; ; Jishi WANG ; ; Qin FANG ; Rui GAO ; Qianying SHI ; ; Hui ZHANG ; Jiangyuan ZHAO ;
Chinese Medical Journal 2014;127(7):1310-1316
BACKGROUNDBone marrow hematopoietic function suppression is one of the most common side effects of chemotherapy. After chemotherapy, the bone marrow structure gets destroyed and the cells died, which might cause the hematopoietic function suppression. Heme oxygenase-1 (HO-1) is a key enzyme of antioxidative metabolism that associates with cell proliferation and resistance to apoptosis. The aim of this study was to restore or resist the bone marrow from the damage of chemotherapy by the HO-1 expression of mouse mesenchymal stem cells (mMSCs) homing to the mice which had the chemotherapy-induced bone marrow suppression.
METHODSOne hundred and sixty female Balb/c mice (6-8-weeks old) were randomly divided into four groups. Each group was performed in 40 mice. The control group was intraperitoneally injected for 5 days and tail intravenously injected on the 6th day with normal saline. The chemotherapy-induced bone marrow suppression was established by intraperitoneally injecting cyclophosphamide (CTX) into the mice which performed as the chemotherapy group. The mMSCs were tail intravenously injected into 40 chemotherapically damaged mice which served as the mMSCs group. The difference between the HO-1 group and the mMSCs group was the injected cells. The HO-1 group was tail intravenously injected into the mMSCs that highly expressed HO-1 which was stimulated by hemin. The expression of HO-1 was analyzed by Western blotting and RT-PCR. Cell proliferation was measured using the 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. Histopathologic examinations were performed 1 week after injection.
RESULTSCompared with the control group, the expression levels of HO-1 mRNA and protein were significantly higher in the HO-1 group (all P < 0.05), even obviously than the mMSCs group. CTX treatment induced apoptosis and inhibited proliferation. After injected, the white blood cell (WBC), red blood cell (RBC) and platelet (PLT) declined fast and down to the bottom at the 7th day. The bone marrow structure was destroyed incomplete. In vitro, the survival rate of cells in chemotherapy group was less than 50% after 24 hours. In contrast, mMSCs could do a favor to the cellular cleavage and proliferation. They slowed down the cell mortality and more than 50% cells survived after 24 hours. The effects of blocking apoptosis and bone marrow recovery could be more effective in the HO-1 group. In the HO-1 group, it had observed that the bone marrow structure became complete and the hemogram closed to normal at 7th day.
CONCLUSIONSHO-1 played an important role in promoting the recovery of CTX-induced hematopoietic damage. We suggest that HO-1 is able to restore the functions of chemotherapy-induced hematopoietic damage.
Animals ; Apoptosis ; drug effects ; Blood Platelets ; drug effects ; Blotting, Western ; Bone Marrow ; drug effects ; enzymology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cyclophosphamide ; toxicity ; Erythrocytes ; drug effects ; Female ; Heme Oxygenase-1 ; genetics ; metabolism ; Leukocytes ; drug effects ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; enzymology ; physiology ; Mice ; Mice, Inbred BALB C ; Reverse Transcriptase Polymerase Chain Reaction
8.Design, synthesis and vasorelaxant activity of R, S-1-(substituted phenyl)-4-3-(naphtha-1-yl-oxy)-2-hydroxypropyl-piperazine derivatives.
Xiao-zhong FU ; Lei TANG ; Mu YUAN ; Jing-shan SHI
Acta Pharmaceutica Sinica 2007;42(7):735-740
According to the results of activity-structure relationship (SAR) studies of alpha1-adrenoceptor antagonists hydantoin-phenylpiperazine and benzimidazo-arypiperazine derivatves, to design and synthesize a series of novel phenylpiperazine alpha1-adrenoceptor antagonists with more potent vasorelaxant activity, active metabolites of naftopidil were used as lead compounds. Ten novel R,S-1-substituted phenyl-4-[3-(naphthal-yl-oxy)-2-hydroxy propyl]-piperazine were designed and synthesized, their vasorelaxant activity was evaluated by calculating inhibition rate of phenylephrine-induced vasocontration of rabbit artery trips. Five compounds exhibited vasorelaxant activity, and compound 16 showed significant vasorelaxant activity in vitro. At 0.01 and 1 micromol x L(-1), its inhibition rates were 7.03% and 22.72%, respectively. This compound possessed ideal vasorelaxant activity in vitro, and would be selected for further anti-hypertension evaluation in vivo. Moreover, by analyzing the primary activity and structure relationship of these compounds, it could be concluded that the SAR results of the reported phenylpiperazine alpha1-adrenoceptor antagonists could be used for reference in designing novel derivatives of naftopidil with optimal pharmacological properties.
Adrenergic alpha-1 Receptor Antagonists
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Animals
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Antihypertensive Agents
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chemical synthesis
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chemistry
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pharmacology
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In Vitro Techniques
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Molecular Structure
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Piperazines
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chemical synthesis
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chemistry
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pharmacology
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Rabbits
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Structure-Activity Relationship
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Vasoconstriction
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drug effects
9.Gene expression of breast cancer resistance protein in adult acute lymphocytic leukemia and its clinical significance.
Xue-Jie JIANG ; Ji-Shi WANG ; Qin FANG
Journal of Experimental Hematology 2008;16(1):31-34
The objective of this study was to investigate the relationship between the expressions of breast cancer resistance protein (BCRP) gene and drug resistance as well as prognosis in adult patients with acute lymphocytic leukemia (ALL). Semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the expression of BCRP gene in 97 adult patients with acute lymphocytic leukemia (ALL) and 30 normal subjects. Beta(2) microglobulin (beta(2)-MG) was used as an internal reference. BCRP/beta(2)-MG ratio >or= 0.48 was defined as BCRP positive. The results showed that the positive ratio of BCRP gene expression in untreated group was 28.7%, in contrast that in refractory and relapsed patients was 51.2%. The first complete remission rates in patients with negative and positive expression were 71.7% and 29.7% respectively. In ALL subtypes BCRP gene expression of L3 type was distinctly higher than that of L(1), L(2) types. In immune types the BCRP gene expression of B-ALL was higher than that of T cell type, especially in mature B cell type with obviously statistical significance (p<0.01). It is concluded that the high expression of BCRP gene may induce clinical drug resistance, and may be an unfavorable factor for prognosis in adult patients with acute lymphocytic leukemia.
ATP Binding Cassette Transporter, Sub-Family G, Member 2
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ATP-Binding Cassette Transporters
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genetics
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metabolism
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Adolescent
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Adult
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Aged
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Drug Resistance, Neoplasm
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genetics
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Female
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Humans
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Male
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Middle Aged
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Neoplasm Proteins
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genetics
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metabolism
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Precursor Cell Lymphoblastic Leukemia-Lymphoma
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genetics
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metabolism
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Prognosis
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Reverse Transcriptase Polymerase Chain Reaction
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Young Adult