In this study, we constructed a yeast consortium surface-display expression system by using Flo1 as an anchor protein. Endoglucanase II (EGII) and cellobiohydrolase II (CBHII) from Trichoderma reesei, and β3-glucosidase 1 (BGLI) from Aspergillus aculeatus were immobilized on Saccharomyces cerevisiae Y5. We constructed the cellulose-displaying expression yeast consortium (Y5/fEGII:Y5/fCBHII:Y5/fBGLI = 1:1:1) and investigated the enzymatic ability and ethanol fermentation. The displayed cellulolytic enzymes was stabile during the 96-h fermentation. The yeast consortium produced 0.77 g/L ethanol from 10 g/L phosphoric acid swollen cellulose (PASC) within 96 h. The yield (in grams of ethanol produced per gram of carbohydrate consumed) was 0.35 g/g, which correspond to 68.6% of the theoretical yield.
Aspergillus ; enzymology ; Cellulase ; genetics ; Cellulose ; metabolism ; Cellulose 1,4-beta-Cellobiosidase ; genetics ; Enzymes, Immobilized ; genetics ; Ethanol ; metabolism ; Fermentation ; Glucosidases ; genetics ; Mannose-Binding Lectins ; metabolism ; Protein Binding ; Saccharomyces cerevisiae ; genetics ; metabolism ; Saccharomyces cerevisiae Proteins ; metabolism ; Trichoderma ; enzymology

Objective To compare the effects of tracheal intubation (TI) and laryngeal mask (LM) during general anesthesia (GA) induction on the α 1-band of quantitative pharmaco-electroencephalography (QPEEG).Methods Fortypatients undergoing GA were randomly divided into two groups:group T included 20 patients who received TI and group L included 20 who received a LM.Parameters like heart rate (HR),mean arterial pressure (MAP),and QPEEG were recorded before anesthesia induction (T0),after induction (T1),and after intubating the cannula or LM (T2).Using power-spectrum analysis,we calculated the power percentage of the α 1-band of QPEEG.Results The HR,MAP,and power percentage of the α 1-band in most areas of the brain were lower at T1 than at T0 (P ＜ 0.05) in both groups.Moreover,the HR,MAP,and α 1-band power percentage were higher at T2 than at T1 (P ＜ 0.05) in group T,whereas they showed no significant change at T2 (P ＞ 0.05) in group L.Conclusion TI is stronger than LM for stimulating the circulatory system.Moreover,TI may cause an increase in the power percentage of the α 1-band of QPEEG.This finding suggests that the α1-band power percentage of QPEEG can be an effective means of monitoring stimulation.

Objective To observe the antidepressant effect of the volatile oil of Myristica in mice and investigate its mechanism.Methods Healthy male Kunming mice were divided into control group,fluoxetine hydrochloride group,high,medium and low dose group,and all of the mice were gavaged for 7 days.The role of antidepressant on the mice were observed with tail suspension tests and forced swimming tests.Then serotonin(5-HT),dopamine(DA) and norepinephrine (NE) in the brain of the mice were determined by enzyme-linked immunosorbent assay.Results Compared with the control group((82.60 ± 24.70)s),the medium dose group could shorten the immobility time of the tail suspension tests ((54.40± 15.87) s),and showed statistical significance (P＜ 0.05).The content of 5-HT,DA,NE in the brain of the medium group were (19.35±2.79) ng/ml,(12.16±0.71)pg/ml,(0.27±0.12) ng/ml,and control group were (14.95±4.83) ng/ml,(11.32±0.95) pg/ml,(0.20±0.11) ng/ml.Compared with the control group,the content of 5-HT of the medium dose group was increased and showed statistical significance(P＜0.05).Conclusion The volatile oil of Myristica fragrans Houtt has antidepressant effect,and may be related to raising the content of 5-HT,DA,NE in the brain of the mice,especially 5-HT.

Objective To compare the effects of different doses of sufentanil on theα1?band of quantitative pharmaco?electroencephalography (QPEEG)during the induction of general anesthesia by tracheal intubation(TI). Methods Forty selected patients under general anesthesia were randomly divided into two groups,with 20 patients per group. Patients in group Ⅰ were administered 0.2μg/kg sufentanil,whereas patients in group Ⅱ were administered 0.3μg/kg sufentanil. Subsequently,the patients were administered 2 mg/kg propofol and 0.15 mg/kg cisatracurium. HR,MAP,and QPEEG were recorded before induction(T0),after induction(T1),and after insertion of the cannula(T2). Using the method of power spectrum analysis,theα1?band power percentage of QPEEG was calculated. Results In comparison with T0,the values of HR,MAP,andα1?band power percentage in most areas of the brain were both decreased at T1(P<0.05). Furthermore,in comparison with T1,the parameters were increased in group Ⅰ at T2(P<0.05),but no significant changes were observed in group Ⅱ (P>0.05). Conclusion The administration of 0.3μg/kg sufentanil during anesthesia induction can effectively depress the cardiovascular response to TI and stabilize theα1?band power per?centage. This suggests that theα1?band power percentage of QPEEG can be an effective means to monitor the depth of sedation.

Objective To observe the interactions between a anti-interleukin-8 monoclonal antibody (anti-IL-8 mAb) carried targeted ultrasound contrast agents and the injured vascular endothelial cells,as well as to explore the role of IL-8 in the formation of atherosclerotic plaques and a new assessing method of vascular endothelial functions.Methods The targeted ultrasound contrast agent was prepared by using a erosslinking agent to couple a anti-IL-8 mAb with SonoVue microbubbles.The interactions between SonoVue microbubbles/the targeted microbubbles and normal/injured endothelial cells were observed under an inverted microscope,respectively.The numbers of endothelial cells and adhered microbubbles were counted under high power magnification.The ratio of microbubbles to endothelial cells was calculated for the quantitative analysis of the interactions.Results In the control group,only a slight amount of original SonoVue microbubbles were bound to normal/injured endothelial cells.In contrast,it was visible under the microscope that the anti-IL-8 mAb carried SonoVue could be bound to endothelial cells,and the number of microbubbles bound to the surface of injured endothelial cells was significantly higher than that bound to the normal endothelial cells.Conclusions The anti-IL-8 mAb carried targeted ultrasound contrast agent could be readily bound to the surface of the injured cells specifically,and thus suggesting a new direction for ultrasonic detection of vascular endothelial injury and the ultrasonic assessment of vascular endothelial functions.

Ischemic stroke is a cerebrovascular disease with high disability rate and mortality. At present,there is no effective treatment to promote the recovery of neurological function after ischemic stroke. Exosomes can not only mediate the communication between cells,but also have the ability to cross the blood-brain barrier,so they have received extensive attention in the treatment of ischemic stroke. Exosomes are modified by bioengineering technologies to prepare engineered exosomes with brain targeting and therapeutic effects,which have been applied in the research and treatment of ischemic stroke,in order to improve the repair of neurological function after stroke,reduce the clinical disability rate and mortality,and improve the survival and quality of life of patients. This article reviews exosomes,the role of exosomes in ischemic stroke,and the preparation of engineered exosomes,and discusses the application prospect of engineered exosomes in the treatment of ischemic stroke,with a view to providing a reference for subsequent research.

Objective To investigate the prevalence of DUOX2 mutations in Chinese patients with congenital hypothyroidism (CH) and to discuss the inheritance pattern of DUOX2 gene.Methods Blood samples were collected from 91 CH children and their genomic DNA was extracted from peripheral blood leukocytes.All exons and exon-intron boundaries of DUOX2 were analyzed by target next-generation sequencing and family trios was established to study the inheritance pattern of DUOX2 gene.Results Fifty-four out of 91 children with CH carried DUOX2 mutation, with a prevalence of 59.34%.Of the 54 CH children, 36 carried DUOX2 biallelic mutations.In all 12 family trios with probands carrying biallelic DUOX2 mutations, the parents carried heterozygous DUOX2 mutations while still showing normal thyroid function, suggesting that CH caused by DUOX2 mutations is inherited in an autosomal recessive manner.Conclusion DUOX2 gene is one of the most frequently mutated genes in Chinese CH patients and its inheritance pattern is an autosomal recessive one.

Objective To investigate the effects of transcutaneous electrical acupoint stimulation (TEAS) on electroencephalography (EEG) in piglets anesthetized with sevoflurane.Methods Twelve piglets,aged three to seven days,weighing 1.5 to 3.5 kg,were randomly divided into 2 groups:TEAS (group T,n =6) and control (group C,n =6).Group T received continuous TEAS at points baihui and tianmen for 30 minutes.Anesthesia was induced with 8.0％ sevoflurane over 3 minutes and maintained with 3.5％ sevoflurane in both groups.The changes were observed on EEG.Results The heart rates (HR) at intubation and extubation were lower in group T than group C (P ＜ 0.05).Compared with group C,the EEG spike frequency was lower in group T during anesthesia induction and maintenance (P ＜ 0.05).Conclusion Sevoflurane can induce EEG spikes in piglets,which can be reduced by TEAS.

Objective To investigate the maternal and fetal outcomes of pregnant women with subclinical hypothyroidism, and clinical observation of thyroxine replacement. Methods From March 2014 to March 2015, the clinical records of 216 women with subclinical hypothyroidism(including 166 cases with thyroxine replacement), and hypothyroidism(n=69)during pregnancy who delivered at our hospital were reviewed. The maternal complications and neonatal outcomes were compared with 406 healthy women who delivered during the same period. Results The age, number of fetus, and morbidity rate of gestational hypertension were without significant differences in those groups. The morbidity of gestational diabetes in subclinical hypothyroidism group and hypothyroidism group were significantly higher than those in control group(13.4%, 17.4% vs 0.2%, P<0.05). The rate of thyroid peroxidase antibody and thyroglobulin antibody positive in subclinical hypothyroidism group and hypothyroidism group were significantly higher than those in control group(26.9%, 23.2% vs 9.9%; 15.7%, 23.2% vs 8.1%, all P<0.05). No matter treated or not treated in subclinical hypothyroidism group, the preterm birth, Apgar score, low birth weight, birth defects, and infant congenital hypothyroidism were without significant differences as compared to the control group(P>0.05). Further compared those between treated and untreated subclinical hypothyroidism, the results were also without significant difference(P>0.05). Conclusions Subclinical hypothyroidism had no significant influence on pregnancy outcomes and perinatal events. Thyroxine replacement in subclinical hypothyroidism pregnant women also had no significant influence on pregnancy outcomes and perinatal events.

Objective:To investigate the role of long non-coding RNA nuclear paraspeckle assembly transcript 1 (Neat1) in pyroptosis of ultraviolet B (UVB)-induced human lens epithelial cells (LECs) and to explore the possible mechanism.Methods:The human lens epithelial cell line HLE-B3 was cultured in vitro, and cells at log phase were exposed to ultraviolet B for 0, 2, 4 and 8 hours, respectively.The expression of cysteine aspartic acid-specific protease-1 (caspase-1), a protein related to pyroptosis, was detected by Western blot.The relative expression level of Neat1 in cells after different irradiation durations was determined by real-time quantitative PCR.Cell viability was determined by the cell counting kit-8 (CCK-8) method to screen the optimal irradiation duration for UVB-induced LECs pyroptosis, which was finally determined to be 4 hours.HLE-B3 cells were divided into negative siRNA transfection group, siRNA Neat1 transfection group, negative siRNA transfection+ irradiation group and siRNA Neat1 transfection+ irradiation group, and were transfected with corresponding reagents for 24 hours.The negative siRNA transfection+ irradiation group and siRNA Neat1 transfection+ irradiation group were irradiated with UVB for 4 hours after transfection.The cell viability was detected by the CCK-8 method.The pyroptosis rate was detected by flow cytometry.The expression levels of caspase-1, gasdermin D (GSDMD) and nod-like receptor protein 3 (NLRP3) proteins were detected by Western blot.The concentration of interleukin (IL)-1β was detected by enzyme-linked immunosorbent assay (ELISA). Ultrastructural changes in HLE-B3 cells were observed under a transmission electron microscope. Results:The grayscale of caspase-1 protein bands increased with the extension of irradiation duration.The relative expression levels of caspase-1 protein at 0, 2, 4 and 8 hours of irradiation were 0.05±0.01, 0.25±0.07, 0.51±0.04 and 0.74±0.02, respectively, with a statistically significant overall difference ( F=168.223, P<0.001), and significant differences were found in paired comparisons (all at P<0.05). With prolonged irradiation, the relative expression level of Neat1 mRNA increased and the cell viability decreased, with statistically significant differences in paired comparisons (all at P<0.05). Compared with negative siRNA transfection group, the cell viability was increased in siRNA Neat1 transfection group and decreased in negative siRNA transfection+ irradiation group, with statistically significant differences (both at P<0.01). Compared with negative siRNA transfection+ irradiation group, the cell viability was increased in siRNA Neat1 transfection+ irradiation group, showing a statistically significant difference ( P<0.05). The pyroptosis rate was significantly lower in negative siRNA transfection group and siRNA Neat1 transfection+ irradiation group than in negative siRNA transfection+ irradiation group, and the differences were statistically significant (both at P<0.01). The relative expression levels of caspase-1, NLRP3 and GSDMD proteins in negative siRNA transfection+ irradiation group were higher than those in negative siRNA transfection group and siRNA Neat1 transfection+ irradiation group and the differences were statistically significant (all at P<0.01). The concentration of IL-1β was significantly higher in negative siRNA transfection+ irradiation group than in negative siRNA transfection group and siRNA Neat1 transfection+ irradiation group, and the differences were statistically significant (all at P<0.05). Cell swelling, formed cell membrane pores, vacuolated cells and fuzzy mitochondrial cristae were seen in negative siRNA transfection+ irradiation group and siRNA Neat1 transfection+ irradiation group by transmission electron microscopy.Compared with negative siRNA transfection+ irradiation group, slighter cell swelling, fewer cell membrane pores and lighter mitochondrial swelling were seen in siRNA Neat1 transfection+ irradiation group. Conclusions:Neat1 is involved in human LECs pyroptosis induced by UVB through the classic pyroptosis pathway mediated by caspase-1.Knockdown of Neat1 can inhibit the pyroptosis of human LECs.