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 ; enzymology ; genetics ; Base Sequence ; Cloning, Molecular ; DNA, Complementary ; Escherichia coli ; Fungal Proteins ; genetics ; metabolism ; Glycosylation ; Hexosyltransferases ; genetics ; metabolism ; Molecular Sequence Data ; Molecular Weight ; Pichia ; Sucrose ; metabolism ; Trisaccharides ; metabolism

Bone Marrow Cells ; Cell Line ; Cell Proliferation ; Humans ; Mesenchymal Stromal Cells ; cytology ; T-Lymphocytes ; cytology ; T-Lymphocytes, Regulatory ; cytology

Cochlea ; metabolism ; Deafness ; genetics ; Humans ; Mutation

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.

Objective To explore the clinical characteristics and management of kidney transplantation in the older adults.Method Forty older kidney recipients (≥60 years old) and 777 younger (18～59 years old) recipients from June 2009 to December 2012 were retrospectively reviewed to evaluate the clinical characteristics and managements.Result Of 40 older recipients,the comorbidities of diabetes and coronary artery disease in older group were higher than in the younger (25％ vs.4.9％,and 32.50％ vs.11.38％).During the first 6 months,7 older patients died,among which 4 died from severe pneumonia,2 from heart failure and 1 from pulmonary embolism.There were 31 deaths in younger group,among which 21 died from severe pneumonia,4 from heart failure,5 from cerebrovascular accident and 2 from pulmonary embolism.The mortality in the older group was higher than in the younger group (17.5％ vs.4.6％).Six-month and 3-year survival rate in the older recipients was lower than the younger recipients (81.56％ vs.95.35％,and 81.56％ vs.94.5％,respectively).Six-month graft survival rate and 3-year survival rate in the older group were also lower than in the younger group (78.75％ vs.92.02％,and 68.82％ vs.85.40％).At the 1st and 2nd year during follow-up,the serum creatinines in the older group were close to those in the younger group,while lower level was observed 3 years after transplantation in the older patients (89.38 ± 11.34 (mol/ L vs.116.57±48.68 (mol/L).Conclusion The patients with ESRD older than 60 years have more preoperative concomitant diseases and worse health status,which contribute to the complex clinical characteristics.More aggressive indication selection,sufficient pre-operative preparation,wellmatched histocompatibility and high quality of donor kidney are the key factors of successful transplantation.Optimized immunosuppressant therapy,delicate perioperative management,preventve and effective treatment of related complications are necessary to promote the survival of recipients and graft in long term.

ObjectiveTo explore the risk factors of obstructive hydrocephalus secondary to intracranial hemorrhage in premature infants.MethodsA total of 304 premature infants were selected who were diagnosed as sever intracranial hemor-rhage (grade III and IV) by cranial bedside ultrasound admitted to our hospital from Jun. 2013 to Sep. 2014. According to wheth-er the obstructive hydrocephalus was followed, all infants were divided into hydrocephalus group (n=59) and non-hydrocephalus group (n=185). The risk factors of obstructive hydrocephalus secondary to intracranial hemorrhage were analyzed and the lateral ventricle size was measured dynamically.ResultsThe univariate analysis showed the factors related with obstructive hydro-cephalus were as follows: gestational age≤32 weeks, birth weight< 1500g, severe asphyxia, cesarean section, RDS, neonatal infection, heart failure, PDA, acidosis, thrombocytopenia, coagulation abnormalities, and intracranial hemorrhage (gradeⅢ orⅣ) (allP<0.05). Multivariate logistic regression analysis showed that acidosis, thrombocytopenia, coagulation abnormalities, gesta-tional age≤ 32 weeks, severe asphyxia, intracranial hemorrhage (gradeⅢ orⅣ) were independent risk factors for obstructive hydrocephalus (OR: 1.76~20.46, allP<0.05). At each time point after birth, the ratio of posterior horn of lateral ventricle was signiifcantly higher in hydrocephalus group than that in non-hydrocephalus group (P<0.05). There were signiifcant differences in the changes of the posterior horn ratio of left or right lateral ventricle with time in hydrocephalus group (P=0.000), increasing at 14 days and reaching the peak at 28 days after birth.ConclusionsThe risk factors for obstructive hydrocephalus secondary to intracranial hemorrhage in neonates are important. Regular and dynamical monitoring of ventricle size by cranial ultrasound is needed in infants with sever intracranial hemorrhage.

Objective To explore the traits of improved polyacrylamide gel electrophoresis (PAGE)-ethidium bromide(EB) staining in the detection of genotype polymorphisms.Methods The methods of PAGE-silver staining,agarose gel electrophoresis (AGE)-EB staining and improved PAGE-EB staining in polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were used to detect the μ-opioid receptor (OPRM1) A118G genotypes in case group (n =167) infants with intracranial hemorrhage (ICH),and control group(n =163) infants without ICH,to conduct a case study analysis.And the application traits of three methods were compared.Results Genotypes of OPRM1 A118G were GG (169 bp),AG (193 bp,169 bp),AA (193 bp).Both the electrophoresis methods of PAGE-silver staining and PAGE-EB could be used to detect the genotypes of OPRM1 A118G clearly in this study.There was no statistically significant difference between the resolutions of DNA fragments (P =0.884).The first method,which had 13 experiment steps,consuming 4-5 hours,involving 12 kinds of chemical reagents,and the pictures were taken with the camera,was complex,with difficult operation,more time consuming; Compared with the first method,the secondmethod was simple,which had 6 test procedures,consuming 2 hours with 8 reagents,and the pictures were taken by using an automatic gel imager.AGE-EB could not be used to detect genotypes of OPRM1 A118G.Conclusions The method of improved PAGE-EB has the advantages of fast,easy operation,and high resolution,which is worthy of wider application.

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.

Objective To report the clinical and magnetic resonance imaging(MRI)features of 5 eases with idiopathic orbital myositis.Methods Four females and one male,aged 27 to 57 years,presented department of neurology in the First Hospital of Peking University in October 2008 to September 2009.The duration of disease Was between 3 months and 4 years.Recurrent course appeared in 3 of them.0rbital MRI Was performed in all of them.After diagnosis they underwent long.term corticosteroid treatment.Results All patients presented ocular pain,asymmetrical and incomplete ophthalmoplegia and mild proptosis.EMG revealed no significant decline in repetitive stimulation.Muscle biopsies of limb muscle were unremarkable.Creatine kinase and thyroid function test were in normal limits.MRI revealed unilateral.focal or difluse enlargement and enhancement of extraocular muscles,involving 1 extraocular muscle in 2 cases,2extraocular muscles in 2 cases,more extraocular muscles in 1 case.No evidence indicated bone destruction or cavernous sinus abnormalities.Five Cases showed improvement and remission after long-term administration of steroids.Conclusion Persistent and asymmetrical ophthalmoplegia is connnon in orbital myositis.Extraocular muscle swelling characterized the MRI changes.

Objective To investigate the possible existence of HBoV in children with acute respiratory infections in Nanjing area and explore its relationship with clinical characteristics.Methods A total of 397 nasopharyngeal secretion samples were collected from children with acute respiratory infection,admitted from July 2009 to June 2010 in Nanjing Children'S Hospital affiliated to Nanjing Medical University,and 50 cases of children without symptoms of respiratory infection were recruited as control group,whose nasopharyngeal secretion samples were also collected.HBoV was determined by real-time fluorescence quantitative PCR.MP and CT were detected by real-time fluorescence quantitative PCR in those HBoV-positive samples.RSV,ADV,IVA,IVB,PIV-1,PIV-2,PIV-3 and hMPV were detected by direct antigen-specific immunofluorescence assays.HBoV NP-1 fragments were amplified and sequenced in 5 HBoV positive samples randomly selected.The results were compared with the known GenBank sequence,and thereby the phylogenetic tree was established.The epidemiological characteristics,clinical presentation and the final clinical diagnosis of HBoV were analyzed according to the clinical data of the HBoV-positive patients.Results Thirty-three HBoV-positive cases were detected by real-time fluorescence quantitative PCR method with a positivity rate of 8. 3% ( 33/397 ). Among the 33 HBoV-positive cases, 19 cases (57.6%) were multiple infections with HBoV and other pathogens, the top three of which were MP (27.3% ,9/33 ),RSV (24.2% , 8/33 ) and PIV-3 ( 12. 1% ,4/33 ). Affected children aged from 7 to 36 months old accounted for 75.8% of the total ( 25/33 ). The measured HBoV NP-1 gene sequences of 5 specimens were consistent,indicating a high homology (99% to 100% ) with the stl, st2 and WHL-1. Conclusions HBoV is one of the pathogens of children's acute respiratory infections in Nanjing. HBoV NP-1 gene is highly conserved,with little variation in different seasons and in different regions and therefore can be used as a marker for real-time fluorescence quantitative PCR and other methods.