Selection of utilization of carbon source in soya oligosaccharide by three strains of S.cerevisiae was studied.The results showed that S.cerevisiae C could selectively utilize sucrose and the residual rate of stachyose and raffinose could be more than 96%.Using yeast extract as nitrogen source,the sucrose could be used up after 36 hours of culture.Further study showed that the content of sucrose in soya oligosaccharide powder was less than 1.3% after fermentation of waste water of soy whey and downstream processing.

Objective To clone and express the alkaline protease AprA , one important virulence factor secreted by Pseudomonas aeruginosa(PAE)in Escherichia coli, to clone and express the inhibitor of AprA (AprI) and its substrate flagellin , and to detect the function of AprA and the inhibitory function of AprI .Methods The genes encoding AprA ,AprI and flagellin gene were amplified respectively by PCR using PAE PAO 1 genome DNA as the template .The expression vec-tors (pET-28a-AprA, pET-28a-AprI and pET-28a-Flagellin) were constructed and transformed into E.coli BL21(DE3) respectively.The recombinant AprA protein was expressed by IPTG induction and purified via denaturing and renaturation. The recombinant AprI and flagellin were expressed and purified by Ni 2+affinity chromatography .The cleavage activities of AprA on flagellin were detected by SDS-PAGE.Results Recombinant AprA , AprI and flagellin protein were expressed and purified .It was demonstrated that AprA cleaved flagellin , which was blocked by AprI .Conclusion Recombinant AprA could cleave its substrates as an alkaline protease , and its inhibitor AprI inhibits the activities of AprA .This study will contribute to further investigations on the role of AprA in the pathogenesis of PAE .

Objective:To clarify the effect of ubiquitination hepatitis B virus core antigen (Ub-HBcAg) on dendritic cells (DC) autophagy, and to explore the mechanism of autophagy in enhancing DC antigen presentation and inducing hepatitis B virus-specific cytotoxic T lymphocyte (CTL) responses.Methods:Ub-HBcAg lentiviral vector (LV-Ub-HBcAg), lentiviral vector-hepatitis B virus core antigen (LV-HBcAg) and no-load plasmid LV (LV) were constructed and packaged. DC2.4 cells were divided into LV-Ub-HBcAg group, LV-HBcAg group and LV group. The blank control group (NC group) was also set. The protein expression of autophagy-related protein P62, microtubule associated protein 1 light chain 3 beta (LC3B), autophagy related 5(ATG5) and Beclin-1 were detected by Western blotting. The expressions of co-stimulatory molecules such as CD86, CD80 and major histocompatibility complex (MHC)-Ⅱ were detected by flow cytometry. Cell counting kit-8 (CCK-8) method was used to detect T lymphocytes proliferation. The non-radioactive lactic acid dehydrogenase (LDH) release method was applied to detect the killing ability of CTL. Statistical analysis was conducted by independent sample t test. Results:The relative protein expressions of LC3B-Ⅱ/LC3B-Ⅰ, Beclin-1 and ATG5 in NC group were 0.445±0.076, 0.522±0.026 and 0.761±0.038, respectively, which were all lower than those in LV-Ub-HBcAg group (0.926±0.021, 0.919±0.016 and 1.451±0.028, respectively). The relative protein expression of P62 in the NC group was higher than that in LV-Ub-HBcAg group ((1.875±0.016) vs (0.647±0.121)). The differences were all statistically significant ( t=6.102, 9.842, 17.490 and 10.590, respectively, all P<0.01). The expressions of CD86 (75.51%), CD80 (83.35%), MHC-Ⅱ (66.66%) in the LV-Ub-HBcAg group were high, and those in the NC group were 8.03%, 7.49%, 0.04%, respectively. The specific CTL killing rate ((65.310±2.091)%) of the LV-Ub-HBcAg group was significantly higher than both NC group ((14.400±0.497)%) and LV-HBcAg group ((54.870±1.443)%), and the differences were both statistically significant ( t=23.690 and 4.111, respectively, both P<0.05). Conclusion:Ub-HBcAg promotes the DC autophagy, up-regulates the expressions of costimulatory molecules on cell surface of DC to induce the maturation and activation, and then stimulates T lymphocyte to induce a stronger specific CTL response under the effort of ubiquitination.

OBJECTIVETo investigate the role of mitochondria in TBMS1-medited apoptosis of human cervical carcinoma HeLa cell line.

METHODMitochondrial transmembrane potentia (deltapsim) was assayed by flow cytometry. Apoptotic induction by TBMS1 was determined by gel electrophoresis of fragmented DNA. Cytochrome c (Cyt c) was detected by Western blotting.

RESULTThe results showed that TBMS1 induced apoptosis in HeLa cells, that TBMS1 decreased deltapsim and facilitated Cyt c release, and that TBMS1 induced apoptosis of HeLa cells dose-and time-dependently in accordance with increase of cytosolic Cyt c. TBMS1 had direct effect on isolated rat mitochondria which induced a time- and dose-dependent release of Cyt c from mitochondria. The results also showed that CsA protected partly HeLa cells from the effect of TBMS1.

CONCLUSIONThe mitochondrial apoptosis pathway has effects on TBMS1 induced human cervical carcinoma HeLa cell line apoptosis.

Animals ; Apoptosis ; drug effects ; Cucurbitaceae ; chemistry ; Cyclosporine ; pharmacology ; Cytochromes c ; metabolism ; Dose-Response Relationship, Drug ; HeLa Cells ; Humans ; Male ; Membrane Potentials ; drug effects ; Mitochondria, Liver ; physiology ; secretion ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Saponins ; administration & dosage ; isolation & purification ; pharmacology ; Triterpenes ; administration & dosage ; isolation & purification ; pharmacology

We studied the alteration of the maize straw apoplast proteins in the process of preservation, and analyzed the effects of apoplast proteins on Penicillum expansum cellulase activities. The results show that: the extractable apoplast proteins are gradually decreased during the preservation of maize straw. Meanwhile, their synergistic effects on P. expensum cellulose are also attenuated. The apoplast proteins extracted from fresh maize straw possess endogenous EG activities, which is unstable and completely vanished after 6 months preservation. The apoplast proteins from the preserved straw exhibit significant synergistic effect on FPA, cotton lyase and beta-glucosidase. The maximal synergistic values are 95.32%, 102.06% and 96.6%, respectively. But interestingly, they inhibit the CMCase activity (max. 49.52%). Apoplast proteins show distinctive synergy with betaG and EG, but have no effect on CBH activity. After eliminating the effect of endogenous EG, the apoplast proteins from fresh maize straw have enhanced synergistic or inhibiting effects on FPA, Cotton lyase, betaG and CMCase than those extracted from the preserved straw. Based on our observation, the apoplast proteins play important roles in regulating the cellulase activities. The detailed analysis of the related mechanisms will greatly benefit the studies of the natural biomaterials hydrolysis.
Cellulase ; metabolism ; Penicillium ; enzymology ; Plant Proteins ; metabolism ; Plant Stems ; metabolism ; Zea mays ; metabolism

A strain KX6,producing thermotolerant endoglucanase,was isolated from compost. The morpholo-gical identification and 16S rRNA sequence analysis showed it belongs to Streptomyces xylophagus. The production and characterization of endoglucanase from Streptomyces xylophagus KX6 was studied. Maximum endoglucanase yield of 0.538 IU/ml was achieved with medium pH8.0,containing CMC2Na 1.0% as carbon resource,soybean meal 1% as nitrogen resource,2% inoculating volume,30% 250 ml triangle flask bulk for medium volume at 40℃ 200r/min shaker for 48h. The endoglucanase exhibited optimum catalytic activity at pH7.0 and 50℃. The enzyme was stable at 50℃,and able to retain 60% of the full activity,when it was incubated at 60℃ for 1h.The enzyme was stable at pH6.0~7.0. All these findings suggest that the enzyme is a thermotolerant neutral endoglucanase.

Objective To carry out mutation analysis of deafness-associated genes for deaf newborns and their parents, and to estimate the recurrence risk for their parents to have deaf descendants.Methods Suspected cases of inherited deafness were identified by neonatal hearing screening and questionnaires. Genomic DNAs of suspected cases and their parents were extracted from their peripheral blood samples . Common deafness-associated genes(i.e. GJB2,SLC26A4 and 12S rRNA genes)were amplified by polymerase chain reaction(PCR),and those PCR products were sequenced for the mutation analysis.Results From 2013 to 2016, 193 cases of deafness were found in neonatal hearing screening,29 cases of suspected as hereditary deafness were screened,and 17 out of 29 cases were found to have mutations in deafness-associated genes(detection rate:58.62%). GJB2 homozygous mutations were identified in two cases and their parents,and the recurrence risk to have deaf descendants was 100%. Four cases of suspected hereditary deafness had GJB2 homozygous mutations,and their parents were both GJB2 mutation carriers. There was one case with SLC26A4 homozygous mutations,and their parents were both SLC26A4 mutation carrier. Two cases were detected to have GJB2 V371 homozygous mutations,and their parents were both GJB2 V371 mutation carriers. For those seven parents carrying deafness-associated mutations above,the recurrence risk of deafness for their descendants was 25%.Conclusion In addition to hearing screening,the genetic diagnosis of deafness-associated genes is helpful to clarify the cause of suspected neonatal hereditary deafness,and can provide objective reproductive counseling and guidance for those deaf parents or parents with deaf children.

Objective To investigate a possible protective effect of sodium ferulate (SF) on monosodium glutamate (MSG)-induced neurotoxicity in adult mice. Methods Sixty mice were randomly divided into control, SF, MSG, and MSG+SF [20,40,80mg/(kg·d)] groups, n=10. The animals in MSG group received intragastric (ig) administration of MSG (2.0g/(kg·d)], the animals in the MSG+SF groups received simultaneously ig administration of MSG [2.0 g/(kg·d)] and intraperitoneal (ip) administration of SF [20,40,80mg/(kg·d)], the animals in SF group received ip administration of SF [40mg/ (kg·d)], and the animals in control group received ig and ip administration of normal saline, respectively, once-daily for 10d. On day 1 after the last ig administration of MSG or (and) SF the behavioural tests (test of Y-maze discrimination learning and open field test) were performed, and on day 4 after the treatment of MSG or (and) SF the histopathology of the animal brains was studied to analyze the MSG-induced functional and morphological changes and the possible protective effect of SF. Results The correct responses of Y-maze test on day 6 after the last administration of MSG and/or SF in MSG-treated group (13.83/20) were significantly less than those in control (16.42/20)(P<0.01), and those in MSG[2.0g/(kg·d)]+SF[40mg/(kg·d)]-treated mice (16.30/20) were close to those in control (P>0.05). Examination of histopathology displayed MSG-treated hippocampal lesions characterized by intracellular edema, degeneration and necrosis of neurons, and hyperplasia, and the hippocampal lesion did not appear in the MSG [2.0g/(kg·d)]+SF[40mg/(kg·d)]-treated mice. Conclusions SF partially countered the behavior disorders and hippocampal lesions induced by MSG; therefor, SF has a potent neuroprotection against MSG-induced neurotoxicity in adult mice.

BACKGROUNDLiver fibrosis normally progresses to cirrhosis and destroys the normal architecture of the liver, resulting in liver dysfunction and irreversible cirrhosis. The aim of this study was to investigate the anti-fibrosis effect and the possible underlying mechanisms of decorin.

METHODSThe mice model of liver fibrosis was induced by intraperitoneal injection of 50% (v/v) of carbon tetrachloride (CCl4) diluted in olive oil (1 ml/kg body weight) once every 2 days for 5 weeks. Three weeks after injecting CCl4 intraperitoneally, mice were randomly divided into normal control with vehicles only (olive oil), mouse model given CCl4 only, and CCl4 plus decorin (DCN, 250 µg/kg). Two weeks later, all the mice were sacrificed and their liver tissues were analyzed for the expressions of genes related to liver fibrosis and under hematoxylin-eosin staining, Masson staining, and immunohistochemical staining of all groups. Aspartate transaminase, alanine transaminase, and total bilirubin of the serum were determined for evaluation of the liver function.

RESULTSExogenous protein decorin could reduce liver fibrosis induced by CCl4 in mice. The degree of fibrosis in the experimental group was alleviated, and the contents of collagen fibers were lower in the experimental group than those of the control group. In addition, expressions of transforming growth factor β1 and α-smooth muscle actin decreased in the experimental group.

CONCLUSIONSTaking liver fibrosis model of mouse as the experimental target and by injecting exogenous protein decorin into the model, we confirmed that decorin could inhibit the expression of proteins related to fibrosis and reduce the formation of liver fibrosis in mice.

Animals ; Carbon Tetrachloride ; toxicity ; Decorin ; therapeutic use ; Immunohistochemistry ; Liver Cirrhosis ; chemically induced ; prevention & control ; Mice ; Transforming Growth Factor beta1 ; metabolism

Objective To understand the incidence and characteristics of nonfatal drowning among primary and middle school students in rural area and to provide basic information for intervention.Methods A rural town was selected and all students from 3th-8th grades,10th grade and 11th grade were studied.All data were collected,using a self-administrated questionnaires which was guided by investigator.Results The overall incidence rate of nonfatal drowning was 5.65%(549/9732)and were 7.69%,5.80%,2.39%for primary,secondary and high school students,respectively.Male students had a higher rate(7.14%)than that of females(4.03%).The incidence rates of non-treated,treated in emergency and under hospitalization were 4.52%,0.77% and 0.35%.The major reasons of drowning were swimming (46.88%),falling into waters(15.67%),diving(13.79%)and rescuing others(6.24%).The proportion of drowning occurred in the afternoon,evening,at noon or in the morning were 59.94%,15.64%,14.77%and 9.65%respectively.The common sites of drowning were river/lake(42.48%),swimming pool(19.56%),reservoir(11.39%)and pond(4.38%).66.76%of the drowning cases were witnessed by other person,and 17.86%were conscious when being removed from waters.Conclusion The incidence of nonfatal drowning among students in rural areas was high,and the natural body of waters was the most common site causing drowning while swimming was the major reason of drowning.Intervention targeting on primary and middle sehool students in rural should be carried out to reduce the incidence.