Lipase from Burkholderia cepacia complex is one of the most versatile biocatalyst and is used widely in many biotechnological application fields including detergent additives, the resolution of racemic compounds, etc. Based on the known whole genomic information of B. cepacia, both ampicillin and kanamycin were added to the TB-T media, the traditional selective media, to screen B. cepacia complex strains from rhizosphere soil samples. The single colonies on the plates with the modified TB-T media were then qualitatively determined the ability to produced the extracellular lipase in the rhodamine B-olive oil agar plates. Thirty-five strains of lipolytic pseudo-B. cepacia complex were isolated and the positive rate of lipolytic bacteria was 65%. Among them, 15 pseudo-B. cepacia complex strains with the tolerance to benzene, n-hexane and n-heptane at the concentration of 10% (V/V) were selected and identified by the recA gene sequence. All of the 15 lipolytic bacteria belonged to the B. cepacia complex.

OBJECTIVETo observe the effect of Rgl treatment on prognosis of alcoholic hepatitis using a rat model.

METHODSFemale Sprague-Dawley rats were radomly divided into four groups:unmodeled control, untreated model, Rgl-treated model, and dexamethasone (DXM)-treated model. The model groups were generated by intragastric injection of alcohol. The unmodeled control group was given an equal dosage of normal saline by the same route. After model establishment, the Rg1 treatment group and the DXM treatment group were administered a 120-hour treatment of Rgl or DXM; the unmodeled controls were administered normal saline on the same schedule. All rats were then fasted for 120 hours and venous blood samples were collected for detection of serum aspartate aminotransferase (AST), alanine transaminase (ALT), total bilirubin (TBil), albumin (Alb), tumor necrosis factor-alpha (TNFat) and interleukin 6 (IL-6). Markers of liver inflammation were measured by immunohistochemistry, western blotting, and real-time quantitative reverse transcription PCR. Fat and apoptosis indices were assessed by hematoxylin-eosin staining and TUNEL assay, respectively. The t-test and F test were used for statistical analyses.

RESULTSThe model group showed remarkably more liver steatosis (over one-third of the tissue) than the unmodeled control group, indicating proper establishment of alcoholic liver disease in the modeled rats. The AST, ALT, TBil, and IL-6 levels were significantly higher in the untreated model group than in the Rgl-treated group and the DXM-treated group. The values were significantly different between the Rg1-treated group and the DXM-treated group:ALT, 69.19+/-8.00 U/L vs.102.88+/-5.16 U/L; TBil, 0.36+/-0.07 µmol/L vs.1.20+/-0.18 µmol/L; IL-6, 126.50+/-6.50 U/ml vs.169.19+/-7.68 U/ml; TNFa, 268.31+/-13.19 µg/L vs.318.94+/-7.87 µg/L (all P less than 0.05). Expression of caspase3 and caspase8 was significantly higher in the model group than in the Rgltreated group and the DXM-treated group (both P<0.05). The apoptosis index was significantly lower in the Rgltreated group and the DXM-treated group than in the model group (both P<0.05). The mRNA and protein expression of caspase3, caspase8 and NF-kB were significantly lower in the Rgl-treated group and the DXM-treated group than in the model group (allP less than 0.05), and the levels of all were significantly lower in the Rgl-treated group cornered to the DXM-treated group (all P<0.05). Conehision In rats with alcoholic hepatitis, Rg1 can significantly relieve pathological injury, improve liver function by blocking the apoptotic pathway, and inhibit release of inflammatory cytokines.

Alanine Transaminase ; Animals ; Aspartate Aminotransferases ; Bilirubin ; Cytokines ; Disease Models, Animal ; Ethanol ; Female ; Ginsenosides ; Hepatitis, Alcoholic ; NF-kappa B ; Rats ; Rats, Sprague-Dawley

Objective To investigate the relationship between multi-slice spiral CT perfusion imaging and tumor angiogenesis and cell cycle protein D1 (cyclinD1) expression in patients with peripheral lung cancer. Methods Seventy-three patients with peripheral lung cancer underwent 16-slice spiral CTperfusion imaging. The CT perfusion imaging were analyzed for time density curve (TDC), perfusion parametric maps and the respective perfusion parameters (BF, BV, PS, PH, and PHpm/PHa). Correlation between the respective perfusion parameters and immunohistochemical findings of MVD measurement and cyclinD1 expression were evaluated. Statistical method used one factor analysis of variance (One-wayANOVA) and Pearson correlation analysis. Results The shape of TDC of three groups of peripheral lung cancers (adenocarcinoma, squamous cell carcinoma, and other type of cancer) was similar. They showed a steeper slope, was obvious increase of nodular HU value [ peak height of TDC is respectively (44. 87±6. 83), (34. 91 ± 8.05), (40. 66±5. 87) HU ], changed little after reaching peak height, became flat at peak, had a platform. Forty-four patients were cyclinD1 positive expression, the peripheral lung cancers with cyclinD1 positive expression showed significantly higher MVD value than that of the peripheral lung cancers with cyclinD1 negative expression [ respectively (33.88± 14. 81), (23. 17±11.66) band/high magnification, P < 0. 01]. The cancers with cyclinD1 positive expression showed significantly higher PH,PHpm/PHa, BF,BV, PS value than those of the cancers with cyelinD1 negative expression[respectively P <0. 05]. MVD was positively correlated with PH, PHpm/PHa, BF, BV, and PS of the three groups ofcancers with cyclinD1 positive expression. Among, the correlation coefficient (r value) of BV, PS, BF was0. 409, 0. 517,0. 503 respectively(all P < 0. 01). MVD was significantly correlated with them. The r valueof PH, PHpm/PHa is 0. 319,0. 324 respectively(all P < 0. 05). MVD was less correlated with them. MVDwas not correlated with PH, PHpm/PHa, BF, BV, and PS of the three groups of cancers with cyclinD1 negative expression. Conclusions Multi-slice spiral CT perfusion imaging moderately correlated with tumorangiogenesis and reflected MVD measurement and cyclinD1 expression. It provided not only a noninvasive method of quantitative assessment for blood flow patterns of peripheral lung cancer but also an applicable diagnostic method for peripheral lung cancer.

Based on previous bioinformational analysis results, two Aspergillus niger lipase (ANL) mutants, ANL-Ser84Gly and ANL-Asp99Pro were constructed to screen ANL mutants with oil-water interface independence. ANL-Ser84Gly still displayed a pronounced interfacial activation, while ANL-Asp99Pro displayed no interfacial activation. The specific activity of ANL-Ser84Gly towards p-nitrophenyl palmitate (-myristate, -laurate and -decanoate) decreased by 29.8% (53.1, 60.1 and 77.1, respectively) than that of ANL, while the specific activity of ANL-Asp99Pro towards p-nitrophenyl palmitate increased by 2.2-fold. The mutation in the hinge region at both sides of the lid domain also destabilized various secondary structure factors of ANL-S84G and ANL-D99P, which resulted in a substantial decrease in thermostability. The achievement to construct oil-water interface-independent ANL mutants would help to further understand lipase interfacial activation mechanism.
Aspergillus niger ; enzymology ; genetics ; Base Sequence ; Enzyme Stability ; Fungal Proteins ; genetics ; metabolism ; Lipase ; genetics ; metabolism ; Molecular Sequence Data ; Mutant Proteins ; genetics ; Oils ; Substrate Specificity ; Water

Carbon dots(CDs)were firstly prepared via a simple pyrolysis route using citric acid as the carbon precursor and glycine as modifier. The obtained CDs were further purified by n-butyl alcohol, which made the particle size more uniform and the fluorescence intensity stronger. When excited at 380 nm, the maximum emission wavelength of the CDs was about 480 nm, with a quantum yield of 47%. Based on fluorescence quenching, a novel method for chloramphenicol assay was developed with glycine-passivated CDs. This method is simple and rapid. A linear relationship between the change of fluorescence intensity and the concentration of chloramphenicol was obtained with a relation coefficient of 0. 999 7. The recovery was in the range of 99% to 101% with a relative standard deviation of 0. 3%, which shows CDs′ potential application in drug detection.

Objective To investigate whether activation of mitochondrial acetal dehydrogenase 2(ALDH2)alleviates hypoxic pulmonary hypertension by regulating the SIRT1/PGC-1α signaling pathway.Methods Thirty 8-week-old C57 BL/6 mice were randomized into control,hypoxia,and hypoxia+Alda-1(an ALDH2 activator)group(n=10),and the mice in the latter two groups,along with 10 ALDH2 knockout(ALDH2-/-)mice,were exposed to hypoxia(10%O2,90%N2)with or without daily intraperitoneal injection of Alda-1 for 4 weeks.The changes in right ventricular function and pressure(RVSP)of the mice were evaluated by echocardiography and right ventricular catheter test,and pulmonary artery pressure was estimated based on RVSP.Pulmonary vascular remodeling,right ventricular injury,myocardial α-SMA expression,distal pulmonary arteriole muscle normalization,right ventricular cross-sectional area,myocardial cell hypertrophy,and right cardiac hypertrophy index were assessed with HE staining,immunofluorescence staining and WGA staining,and the expressions of ALDH2,SIRT1,PGC-1α,P16INK4A and P21CIP1 were detected.In pulmonary artery smooth muscle cells with hypoxic exposure,the effect of Alda-1 and EX527 on cell senescence and protein expressions was evaluated using β-galactose staining and Western blotting.Results The wild-type mice with hypoxic exposure showed significantly increased RVSP,right ventricular free wall thickness and myocardial expressions of P16INK4A and P21CIP1,which were effectively lowered by treatment with Alda-1 but further increased in ALDH2-/-mice.In cultured pulmonary artery smooth muscle cells,hypoxic exposure significantly increased senescent cell percentage and cellular expressions of P16INK4A and P21CIP1,which were all lowered by treatment with Alda-1,but its effect was obviously attenuated by EX527 treatment.Conclusion ALDH2 alleviates hypoxia-induced senescence of pulmonary artery smooth muscle cells by upregulating the SIRT1/PGC-1α signaling pathway to alleviate pulmonary hypertension in mice.

Objective To investigate whether activation of mitochondrial acetal dehydrogenase 2(ALDH2)alleviates hypoxic pulmonary hypertension by regulating the SIRT1/PGC-1α signaling pathway.Methods Thirty 8-week-old C57 BL/6 mice were randomized into control,hypoxia,and hypoxia+Alda-1(an ALDH2 activator)group(n=10),and the mice in the latter two groups,along with 10 ALDH2 knockout(ALDH2-/-)mice,were exposed to hypoxia(10%O2,90%N2)with or without daily intraperitoneal injection of Alda-1 for 4 weeks.The changes in right ventricular function and pressure(RVSP)of the mice were evaluated by echocardiography and right ventricular catheter test,and pulmonary artery pressure was estimated based on RVSP.Pulmonary vascular remodeling,right ventricular injury,myocardial α-SMA expression,distal pulmonary arteriole muscle normalization,right ventricular cross-sectional area,myocardial cell hypertrophy,and right cardiac hypertrophy index were assessed with HE staining,immunofluorescence staining and WGA staining,and the expressions of ALDH2,SIRT1,PGC-1α,P16INK4A and P21CIP1 were detected.In pulmonary artery smooth muscle cells with hypoxic exposure,the effect of Alda-1 and EX527 on cell senescence and protein expressions was evaluated using β-galactose staining and Western blotting.Results The wild-type mice with hypoxic exposure showed significantly increased RVSP,right ventricular free wall thickness and myocardial expressions of P16INK4A and P21CIP1,which were effectively lowered by treatment with Alda-1 but further increased in ALDH2-/-mice.In cultured pulmonary artery smooth muscle cells,hypoxic exposure significantly increased senescent cell percentage and cellular expressions of P16INK4A and P21CIP1,which were all lowered by treatment with Alda-1,but its effect was obviously attenuated by EX527 treatment.Conclusion ALDH2 alleviates hypoxia-induced senescence of pulmonary artery smooth muscle cells by upregulating the SIRT1/PGC-1α signaling pathway to alleviate pulmonary hypertension in mice.

Objective 　To investigate the optimized range of body mass index (BMI) selection for patients undergoing 70 kV high-pitch dual-source coronary CT angiography (CCTA) on the third-generation dual-source CT (DSCT). Methods 　Patients undergoing prospective high-pitch ultra-low contrast media (CM) CCTA on the third-generation DSCT using the automatic tube voltage selection at 70 kV were included and divided into three groups:group A,with BMI≤24 kg/m;group B,with 24 kg/m0.05). Subjective image quality in group A(Z=2.91,P=0.004) and B(Z=2.27,P=0.021) were both significantly better than that in group C. Conclusion 　The ultra-low tube voltage (70 kV) combined with ultra-low CM CCTA protocol on third-generation high-pitch DSCT may be better for patients with BMI<28 kg/m2 than those with BMI ≥28 kg/min China.
Body Mass Index ; China ; Computed Tomography Angiography ; methods ; Contrast Media ; Heart Rate ; Humans ; Prospective Studies ; Radiation Dosage ; Radiographic Image Interpretation, Computer-Assisted ; Signal-To-Noise Ratio