ABSTRACT
Self-assembling amphipathic peptides (SAPs) have alternating hydrophilic and hydrophobic residues and can affect the thermal stabilities and catalytic properties of the fused enzymes. In this study, a novel multifunctional tag, S1vw (HNANARARHNANARARHNANARARHNARARAR) was developed to modify fused enzymes. After fusing S1vw at the enzymes/proteins N-terminus through a PT-linker, the crude enzymatic activities of polygalacturonate lyase and lipoxygenase were enhanced 3.1- and 1.89-fold, respectively, compared to the wild-type proteins. The relative fluorescence intensity of the green fluorescent protein was enhanced 16.22-fold. All the three S1vw fusions could be purified by nickel column with high purities and acceptable recovery rates. Moreover, S1vw also induced the thermostabilities enhancement of the fusions, with polygalacturonate lyase and lipoxygenase fusions exhibiting 2.16- and 3.2-fold increase compared with the corresponding wild-type, respectively. In addition, S1vw could enhance the production yield of green fluorescent protein in Escherichia coli and Bacillus subtilis while the production of GFP and its S1vw fusion changed slightly in Pichia pastoris. These results indicated that S1vw could be used as a multifunctional tag to benefit the production, thermal stability and purification of the fusion protein in prokaryotic expression system.
Subject(s)
Escherichia coli , Green Fluorescent Proteins , Hydrophobic and Hydrophilic Interactions , Peptides , Pichia , Recombinant Fusion Proteins , MetabolismABSTRACT
BACKGROUND:Isoflurane cannot only induce a wide range of large neuronal apoptosis, but also inhibit hippocampal neurogenesis in neonatal rats, thereby resulting in hippocampus-dependent learning and memory defects. OBJECTIVE:To investigate the isoflurane effect on proliferation and differentiation of the hippocampal neural stem cels. METHODS:Twenty-six Sprague-Dawley rats were randomly divided into air group and isoflurane group (n=13 per group). Rats in the isoflurane group were subjected to 2.5% isoflurane inhalation for 3 minutes folowed by 1.5% isoflurane inhalation for 4 hours. Rats in the air group only breathed in air. After the intervention, blood glucose and arterial blood gas changes were detected in the two groups. Additionaly, rats in the two groups were given intraperitoneal injection of 5-bromodeoxyuridine before and after intervention. At 24 hours after the last injection of 5-bromodeoxyuridine, brain tissues were taken to make frozen sections for immunofluorescence staining. RESULTS AND CONCLUSION:There were no significant difference in pH, PaO2, PaCO2, HCO3, BE and SaO2 levels between the two groups (P> 0.05). Compared with the air group, the number of BrdU+ cels was significantly less in the isoflurane group (P < 0.05), while the number of NeuroD+/BrdU+ cels was significantly higher in the isoflurane group (P < 0.05). The incidence of adverse reactions was 23% in the isoflurane group, which was significantly higher than that in the air group (7.7%;P < 0.05). These findings indicate that isoflurane can inhibit the proliferation of neural stem cels in the hippocampal dentate gyrus, and promote their differentiation into neurons.
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OBJECTIVE:To observe the effects of dexmedetomidine hydrochloride on extubation stress response in elderly pa-tients during general anesthesia recovery period. METHODS:80 patients underwent general anesthesia of abdominal surgery were randomly divided into observation group and control group,with 40 cases in each group. Both groups received routine anesthesia before surgery and during surgery. 10 min before the end of surgery,observation group was given intravenous pump of dexmedeto-midine hydrochloride diluted to 10 ml 0.5 μg/kg,and control group was given constant volume of 10 ml 0.9% Sodium chloride so-lution. HR,MAP and SaO2 levels of 2 groups were compared before the induction of anesthesia (T0),at the moment of intrave-nous injection of dexmedetomidine hydrochloride or 0.9%Sodium chloride solution(T1),at the moment immediately before extuba-tion(T2),at the moment of extubation(T3),1 min after extubation(T4),5 min after extubation(T5)and 10 min after extubation (T6). Ramsay sedation score of 2 groups were compared at T0,T2,T4,T5 and T6;cough score,restlessness score,awake time(t1) and extubation time(t2)were also compared between 2 groups. RESULTS:Compared with at T0,the fluctuation of HR and MAR in 2 groups decreased significantly and became stable gradually at T4,T5 and T6,and HR and MAP of observation group at T1-T6 were significartly lower than those of control group,with statistical significance(P0.05). There was statistical significance in Ramsay sedation score between 2 groups at T2,T4,T5 and T6 (P<0.05). The cough score and restlessness score of observation group were significantly lower than those of control group,and t1 and t2 were significantly shorter than those of control group,with statistical significance(P<0.05). CONCLUSIONS:Dexmedetomidine hydro-chloride can relieve extubation stress reaction in elderly patients during general anesthesia recovery period with good safety.
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Background and purpose:Clinical data show that Endostar, a recombinant human endostatin, has the therapeutic beneift for patients with non-small cell lung cancer (NSCLC) while combined with chemotherapy or ra-diotherapy. However, the microenvironment changes induced by Endostar monotherapy in NSCLC is not yet clear. The purpose of this study was to prospectively study tumor vascular effects of Endostar monotherapy in patients with locally advanced or advanced NSCLC by dynamic contrast-enhanced perfusion computed tomography (CT perfusion, CT-p). Methods:Previously untreated patients with histologically or cytologically conifrmed locally advanced or advanced NSCLC were eligible. All patients received daily Endostar (7.5 mg?m2) for 14 days. CT-p scans were acquired at the baseline and post-treatment. CT-p parameters, such as blood lfow (BF), blood volume (BV) and permeability surface PS (area product), were measured in all patients.Results:Of all 7 patients enrolled, four were staged asⅢB and three as stageⅣ (2 with malignant pleural effusion, 1 with brain metastasis). The median BF, BV and PS values of baseline and post-treatment were 27.1/48.9 mL/100 mL/min, 86.8/84.8 mL/100 mL and 45.0/54.0 mL/100 mL/min, respectively. After administration of Endostar for 14 days , BF showed a signiifcant increase compared with that at baseline (P=0.028), whereas no signiifcant changes were found in BV (P=0.398) and PS (P=0.237) values.Conclusion:Our results suggest that Endostar monotherapy induces a signiifcant increase in BF whereas no signiifcant difference in BV and PS.
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Background and purpose:We investigated whether lfuorine-18 lfuorodeoxyglucose (18F-FDG) maximal standard uptake value (SUVmax) of the primary tumor (SUV-T), SUVmax of the regional lymph nodes (SUV-N) or the overall loco-regional lesion SUVmax (SUV-TOTAL) was related to survival of patients with stage Ⅲ non-small cell lung cancer (NSCLC) who received Cetuximab and combined definitive chemoradiotherpay. Methods:From September 2009 to July 2012, seventeen patients with unresectable stageⅢNSCLC receiving cetuximab with cisplatin/vinorelbine (NP) followed by concomitant NP and intensity-modulated radiotherapy (IMRT) at the Fudan University Shanghai Cancer Center were enrolled onto a prospectively study. All patients received positron emission tomography/computerized tomography (PET/CT) scans within 2 weeks before enrolment. Univariate analysis were used to assess the correlation between SUV-T, SUV-N, SUV-TOTAL, gender, age, histology, tumour-node-metastasis (TNM) stage, performance status (PS) as well as smoking status and survival. The factors which showed statistical signiifcance entered into multivariate Cox-regression model. Survival functions of different populations were estimated by Kaplan-Meier method and compared by Log-rank test. Results:In the univariate analysis, SUV-T, SUV-N, SUV-TOTAL, PS and smoking status were prognostic factors. The best cut-off values for SUV-T, SUV-N and SUV-TOTAL were 11, 11 and 20, respectively. Multivariate analysis revealed that SUV-TOTAL (P=0.012), SUV-T (P=0.025), and SUV-N (P=0.033) were independent predictors of survival with hazard ratio (HR) of 14.7, 11.2, and 6.2, respectively. Conclusion:Local, regional and locoregional maximal SUVs deifned by 18F-FDG PET-CT scanning may have a strong correlation with survival in this patients setting, which merits further study.
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Radiation therapy or chemoradiation will be used much more commonly in the treatment of patients in postoperative gastric cancer . We have reviewed the indication, toxicity, radiation dose, radiation field in postradiotherapy of gastric cancer patients.