2.Molecular cloning and over-expression of a fructosyltransferase from Aspergillus niger QU10.
Guoqing ZHANG ; Jing YANG ; Jiaji SHI ; Shijun QIAN ; Yapeng CHAO
Chinese Journal of Biotechnology 2015;31(4):512-522
The main commercial production of fructooligosaccharides (FOS) comes from enzymatic transformation using sucrose as substrate by microbial enzyme fructosyltransferase. A fructosyltransferase genomic DNA was isolated from Aspergillus niger QU10 by PCR. The nucleotide sequence showed a 1 941 bp size, and has been submitted to GenBank (KF699529). The cDNA of the fructosyltransferase, containing an open reading frame of 1 887 bp, was further cloned by RT-PCR. The fructosyltransferase gene from Aspergillus niger was functionally expressed both in Escherichia coli and Pichia pastoris GS 115. The highest activity value for the construction with the α-factor signal peptide reached 431 U/mL after 3 days of incubation. The recombinant enzyme is extensively glycosylated, and the active form is probably represented by a homodimer with an apparent molecular mass of 200 kDa as judged from mobility in seminative PAGE gels. The extracellular recombinant enzyme converted sucrose mostly to FOS, mainly 1-kestose and nystose, liberating glucose. FOS reached a maximal value and represented about 58% of total sugars present in the reaction mixture after 4 h reaction. The results suggest that the availability of recombinant Pichia pastoris as a new source of a FOS-producing enzyme might result of biotechnology interest for industrial application.
Aspergillus niger
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enzymology
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genetics
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Base Sequence
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Cloning, Molecular
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DNA, Complementary
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Escherichia coli
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Fungal Proteins
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genetics
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metabolism
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Glycosylation
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Hexosyltransferases
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genetics
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metabolism
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Molecular Sequence Data
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Molecular Weight
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Pichia
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Sucrose
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metabolism
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Trisaccharides
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metabolism
3.Optimization of the honey-fried processing for Radix et Rhizoma Glycyrrhizae by orthogonal design
Qian ZHOU ; Tai ZHANG ; Dianhua SHI ; Lili SUN
Chinese Traditional Patent Medicine 1992;0(03):-
AIM:To choose the optimal processing condition for stir-frying Radix et Rhizoma Glycyrrhizae with honey.METHODS:The contents of glycyrrhizic acid and liquiritin and macroscopical identification were selected as indexes according to orthogonal design L9(34),four factors including the proportion of processed honey and water,soaking moistening time,drying temperature,and drying time.RESULTS:Optional processing conditions consisted of using two part processed honey to one part water as mixture,30 min soaring moistening time,oven temperature rose to 130 ?C and keeping for 20 min.CONCLUSION:The above processing can make the contents of glycyrrhizic acid liquiritin maintaining more 2.33%(RSD 0.43%) and 0.9%(RSD 1.10%),the optimal honey-fried processing technology for Radix et Rhizoma Glycyrrhizae is reasonable.
4.Effects of urinastatin on the expressions of NF-?B and TNF-?, liver transplantation in patients
Shaobo ZHANG ; Qian HE ; Liang GE ; Jianshe SHI ; Gengjie WANG
Chinese Journal of Current Advances in General Surgery 1999;0(02):-
Objective: To investigate the effect of urinastatin on the expression levels of NF-?B and TNF-? liver transplantation in patients. Methods: Thirty-six patients of late stage hepatic cirrhosis were randomly divided into two eaquel groups: the experiment group was administrated by urinastatin 300 000 U and saline 10 mL via venous injection during liver transplantation, the control group was administrated 10 mL saline by venous injection at the same time. The blood samples were harvestd at 1, 2, 4 and 6 h after the blood recovery of donated liver. The nuclear factor-?B (NF-?B) P65 relative level of blood was detected by Western blot analysis and the TNF-? level of blood was detected by double antibody Sandwich enzyme linked immunosorbent assay. Serum ALT and AST were also measured. Results: The expression level of serum NF-?B p65, TNF-? in the experiment group were lower than those in the control group, and the difference was significant(P
6.Optimization of the honey-fried processing for Radix et Rhizoma Glycyrrhizae by orthogonal design
Qian ZHOU ; Tai ZHANG ; Dianhua SHI ; Lili SUN
Chinese Traditional Patent Medicine 2010;(3):447-450
AIM:To choose the optimal processing condition for stir-frying Radix et Rhizoma Glycyrrhizae with honey.METHODS:The contents of glycyrrhizic acid and liquiritin and macroscopical identification were selected as indexes according to orthogonal design L_9(3~4),four factors including the proportion of processed honey and water,soaking moistening time,drying temperature,and drying time.RESULTS:Optional processing conditions consisted of using two part processed honey to one part water as mixture,30 min soaring moistening time,oven temperature rose to 130 ℃ and keeping for 20 min.CONCLUSION:The above processing can make the contents of glycyrrhizic acid liquiritin maintaining more 2.33%(RSD 0.43%)and 0.9%(RSD 1.10%),the optimal honey-fried processing technology for Radix et Rhizoma Glycyrrhizae is reasonable.
7.Expression of type Ⅰ interferon in monocyte-derived dendritic cells after Toll like receptor 3 triggered in patients with chronic hepatitis B
Yujie ZHANG ; Guangfeng SHI ; Qian LI ; Ning LI ; Mingquan CHEN
Chinese Journal of Infectious Diseases 2009;27(6):343-347
Objective To detect the expression of type Ⅰ interferon in monocyte-derived dendritic cells(MoDCs)after Toll like receptor(TLR)3 triggered in patients with chronic hepatitis B(CHB),and to evaluate immune responses of CHB patients and its roles in the mechanisms of persistent infection of hepatitis B virus(HBV)and chronicity of hepatitis.Methods Peripheral blood mononuclear cells(PBMCs)were isolated and purified using magnetic beads(plasma was saved simultaneously)from 26 CHB patients and 18 healthy volunteers(HV).Dendritic cells(DCs)were induced and proliferated in a culture medium with recombinant human granulocyte macrophage colony stimulating factor(rhGM-CSF)and recombinant human interleukin(rhIL-4).EX3s were stimulated with Poly Ⅰ:C and the supernatants were collected at 0 h and 24 h after stimulation.Type Ⅰ interferon(IFN-α and IFN-β)in plasma and supernatants were examined by enzyme linked immunosorbent assay (ELISA).Results The levels of type Ⅰ interferon in plasma were not significantly different in groups of HV and CH B.IFN-α and IFN-β expressions in supernatants before Poly Ⅰ:C stimulation were(80.00±16.15)ng/L,(36.39±13.90)ng/L in CHB group and(76.76±15.90)ng/L,(37.14±13.68)ng/L in HV group,respectively.And there were no statistical differences between two groups(t=1.651,t=0.178;both P>0.05).IFN-α expressions in supernatants at 24 h after stimulation in two groups were both higher than those before stimulation(at 0 h),but there were no statistical differences(t=1.534,t=1.243;both P>0.05).IFN-β expressions in supernatants at 24 h after stimulation in HV group was(54.57±16.80)ng/L,which was significantly higher than that at 0 h(37.14±13.68)ng/L(t=4.061,P<0.05).However,there was no significant difference at 24 h than tht at 0 h in CHB group(t=1.796,P>0.05).At 24 h after stimulation.IFN-β level was(54.57±16.80)ng/L in HV group,which was significantly higher than that[(41.64±12.57)ng/L]in CHB group(t=2.921,P<0.05).Conclusions Functions of MoDCs from CHB patients are impaired and MoDCs could not express type Ⅰ interferon normally.Expression of type Ⅰ interferon after TLR3 triggered in CHB patients is mainly IFN-β.
8.Clinical characteristics and management of acute myocardial infarction after kidney transplantation
Liping CHEN ; Xiaoling ZHANG ; Li XIAO ; Yeyong QIAN ; Bingyi SHI
Chinese Journal of Organ Transplantation 2013;34(10):604-606
Objective To explore the clinical characteristics and management of acute myocardial infarction (AMI) early after kidney transplantation (<3 months).Method Five cases of AMI early posttransplantation among 122 kidney transplant recipients from June 2011 to December 2012 were retrospectively reviewed.Results Of 5 AMI patients,there were 2 cases within one week postoperatively,one case at 11 th day postoperation,and the other two at 29th day and 46th day after operation respectively.Acute left heart failure was complicated in 3 cases within first two weeks.All the AMI patients had elevated TnⅠ levels which declined subsequently.The climax of TnⅠ levels in all the 5 AMI patients were above 5 ng/mL,and more than 20 ng/mL in two AMI patients within one week.Given by symptomatic and supportive treatment,antiplatelet and anticoagulation therapies and cardioprotective medications,all the five AMI patients were improved.Low molecular heparin was additionally administrated to the 2 cases within first week according to the severe conditions.New emerged small volume of hematocele was proved by ultrasound after 3 days and low molecular heparin was ceased.All the 5 patients survived and neither thrombolysis nor percutaneous coronary intervention therapy was given to them.Conclusion In addition to general prevention against AMI in kidney recipients with high risk factors,managing anemia and hypertensiorn,and improving graft function and systematic status are also important to decrease the risk of AMI.Moreover,cardioprotective therapy including antiplatelet therapies,beta-blockers,angiotensin-converting enzyme inhibitors (ACEI)/angiotensin-2 receptor blockers and statins,which are recommended to the general population with AMI,will also profit to the kidney transplant recipients with AMI.However,aggressive intervention therapies might be more prudent to be used in this population.
9.Follow-up study of discharged patients infected with novel recombinant avian-origin influenza A H7N9
Ying ZHU ; Zhiyong ZHANG ; Yuxin SHI ; Qingle WANG ; Qian MA
Chinese Journal of Radiology 2013;47(9):786-789
Objective To analyze the laboratory results and chest CT appearances of novel recombinant avian-origin influenza A (H7N9) in discharged patients for understanding the imaging changes.Methods The clinical and imaging data of 4 patients with novel recombinant avian-origin influenza A (H7N9) were collected.The imaging changes and laboratory results were analyzed.Results All patients underwent chest CT examination 2 to 5 times from admission to discharge.The lesions primarily presented as ground-glass opacity (GGO) and mainly located in the inferior lobe of the left lung in 3 cases,in the inferior lobe of the right lung in 1 case,alternating between absorption and progress.GGO was absorbed obviously in about 3 days after treatment and consolidation was progressed after treatment.Consolidation absorption was observed with continue treatment in 1 case.The pulmonary lesions were absorbed obviously in 1 case 7 days after discharge.Synchronous reduction of the absolute values of CD3,CD8,CD4,CD45 were observed in 3 cases and the ratio of CD4/CD8 was normal.Conclusions (1) The areas of pulmonary involvement are large and variable in H7N9.The lesion area and imaging patterns are important for disease prognosis.(2) H7N9 virus infection may causes immunosuppression,decrease of CRP value can predict the improvement of the disease.
10.The Research Progress of Nurse Professionalism and Its Influencing Factors
Qian WU ; Xiao SUN ; Yan SHI ; Shuying ZHANG
Chinese Medical Ethics 2015;(4):491-494
This paper expounds the concept and connotation of the nurse professionalism, introduces the re-search status of foreign nurses professionalism assessment tools,discusses the influence factors of nurse professional-ism,including:personal factors, organizational factors and occupational factors.It also puts forward following sugges-tions:constructing the nurse professionalism of theoretical system and evaluation methods, advocating the spirit of nurses professionalism cultivation,creating a health care professional environment,providing the reference the devel-opment of our country's theory and improve the nurse professionalism.