The β-Glucuronidase gene (sbGUS) cDNA firstly from Scutellari abaicalensis leaf was cloned by RT-PCR, with GenBank accession number KR364726. The full length cDNA of sbGUS was 1 584 bp with an open reading frame (ORF), encoding an unstable protein with 527 amino acids. The bioinformatic analysis showed that the sbGUS encoding protein had isoelectric point (pI) of 5.55 and a calculated molecular weight about 58.724 8 kDa, with a transmembrane regions and signal peptide, had conserved domains of glycoside hydrolase super family and unintegrated trans-glycosidase catalytic structure. In the secondary structure, the percentage of alpha helix, extended strand, β-extended and random coil were 25.62%, 28.84%, 13.28% and 32.26%, respectively. The homologous analysis indicated the nucleotide sequence 98.93% similarity and the amino acid sequence 98.29% similarity with S. baicalensis (BAA97804.1), in the nine positions were different. The expression level of sGUS was the highest in root based on a real-time PCR analysis, followed by flower and stem, and the lowest was in stem. The results provide a foundation for exploring the molecular function of sbGUS involved in baicalcin biosynthesis based on synthetic biology approach in S. baicalensis plants.
Amino Acid Sequence ; Base Sequence ; Cloning, Molecular ; Computational Biology ; Glucuronidase ; chemistry ; genetics ; metabolism ; Molecular Sequence Data ; Open Reading Frames ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Protein Structure, Secondary ; Scutellaria baicalensis ; chemistry ; enzymology ; genetics ; Sequence Alignment

Objective To study the acute inhalation toxicity of 50% Luowei,a plant molluscacide,in rats. Methods Twenty adult Wistar rats,half male and half female,were given of 5000 mg/m3 of 50% Luowei through a dynamic inhalation de?vice and the death and recovery of the rats were observed within 14 days,and LC50 was drawn. Results After exposure,the ac?tivities of the rats decreased and a few individual animals had scratching symptom,but the symptom disappeared after the expo?sure. No animals died during the whole observation period. Therefore,the acute inhalation LC50 was greater than 5000 mg/m3 for rats,and Luowei belonged to low toxicity level. Conclusions 50% Luowei will not cause animal death if it is inhaled into ani?mal body. However,people still need to wear protective equipment in production and use.

A test was conducted to examine the effect of several electron donors such as glucose, sodium acetate, Fe0, Fe0+glucose and Fe0+sodium acetate on reductive dechlorination of 2,4-dichlorophenol (2,4-DCP) through inoculating the unacclimated anaerobic mixed bacteria. The optimum condition and sus-tainability of Fe0 as electron donor was also been discussed. The results showed that, Fe0+glucose enhanced the dechlorination of contaminant effectively compared to glucose. Sodium acetate, Fe0 and Fe0+sodium acetate were all effective electron donors and Fe0 was the optimum, the optimum initial pH was 8.0 and quantity of added Fe0 was 2.0 g/L. 4-CP was the mainly intermediate product for 2,4-DCP dechlorination. Fe0 could support the electron for reductive dechlorination of 2,4-DCP continuously. In contrast, when so-dium acetate as electron donor, the effect of dechlorination was inferior to Fe0 with the consumption of sodium acetate.

OBJECTIVETo investigate the role of endoplasmic reticulum stress in the apoptosis of testicular germ cells in hyperlipidemic rats.

METHODSWe randomly assigned 42 four-week-old male Wistar rats into a normal control group (n = 12) and a high-fat group (n = 30) to be fed on a normal diet and a high-fat diet, respectively, for 10 weeks. Then we measured the concentrations of triglyceride (TG) and total cholesterol (TC) in the serum using an automatic biochemistry analyzer, detected the apoptosis of testicular germ cells by TUNEL staining, and determined the protein and mRNA expressions of GRP78 and. caspase-12 in the testis tissue by immunohistochemistry and RT-PCR, respectively.

RESULTSThe concentrations of TG and TC were significantly increased in the animals of the high-fat group ([3.00 ± 0.92] and [3.04 ± 0.39] mmol/L) as compared with the control rats ([1.43 ± 0.41] and [1.55 ± 0.23] mmol/L) (P < 0.01), and so was the apoptosis index of the testicular germ cells ([37.17 ± 2.74]% vs [5.16 ± 0.81]%, P < 0.01). The high-fat group, in comparison with the control, also showed remarkably upregulated protein and mRNA expressions of GRP78 (0.32 ± 0.03 and 0.86 ± 0.05 vs 0.19 ± 0.01 and 0.37 ± 0.03, P < 0.01) and caspase-12 (0.34 ± 0.02 and 0.87 ± 0.01 vs 0.12 ± 0.01 and 0.34 ± 0.03, P < 0.01) in the testis tissue.

CONCLUSIONThe apoptosis of testicular germ cells is increased in hyperlipidemic rats, which may be attributed to endoplasmic reticulum stress.

Animals ; Apoptosis ; physiology ; Caspase 12 ; metabolism ; Cholesterol ; blood ; Diet, High-Fat ; adverse effects ; Endoplasmic Reticulum Stress ; physiology ; Heat-Shock Proteins ; metabolism ; In Situ Nick-End Labeling ; Male ; Random Allocation ; Rats ; Rats, Wistar ; Spermatozoa ; pathology ; Staining and Labeling ; Testis ; metabolism ; Transcriptional Activation ; Triglycerides ; blood ; Up-Regulation

Objective: To investigate the free radical scavenging activities of the extract from pomegranate peel and compare with that from juice. Methods: Free radicals (-2O, 稯H and ROO? scavenging activities were investigated in different special chemical systems. Its inhibition activity on LDL oxidation was studied with LDL oxidation model in vitro. Results:Both extracts had strong abilities to scavenge -2O, 稯H and ROO?radicals and prevent LDL oxidation in dose-dependent manner. The extract from peel showed higher activities than that from juice. Conclusion: The natural antioxidants in the extracts of pomegranate peel or juice can effectively scavenge -2O, 稯H and ROO?radicals, and prevent LDL oxidation significantly, especially much more in the former.

Objective: To investigate the effects of pomegranate peel extract on antioxidant capacity and lipid metabolism in C57BL/6J mice fed high fat diet in comparison with pomegranate juice extract. Method: The hyperlipidemic model was developed by feeding C57BL/6J mice a high fat diet. The effects of pomegranate peel extract (supplemented in drinking water) on serum FRAP value, paraoxonase(PON), GSH-Px and SOD activities as well as serum and hepatic lipids contents were observed. The morphologic change of aortic walls was also examined. Results: The antioxidant capacity and lipid metabolism were improved by supplementation of pomegranate peel extract in hyperlipidemic mice. The pathologic changes manifested in aortic walls were also inhibited. The pomegranate juice extract presented similar effects. Conclusion: The pomegranate peel extract inhibited the early process of atherosclerosis. The possible mechanisms were related to the improved antioxidant capacity and lipid metabolism.

OBJECTIVETo prospectively compare the rates of gastroesophageal variceal rebleeding in patients underwent TIPS alone and TIPS combined with embolization of gastric coronary veins.

METHODSAccording to the bleeding state within one week before the shunt placement, 122 patients with hepatic cirrhosis indicated for the secondary prevention of gastroesophageal variceal rebleeding were allocated to the shunt group (n = 44, treated with TIPS alone) and the shunt plus embolization group (n = 78, treated with TIPS combined with embolization of gastric coronary veins). All the patients were followed up for 1 year, and the 1-year cumulative rates of rebleeding, shunt patency and mortality were compared.

RESULTSThe basic characteristics of patients in the two groups were comparable (P is more than 0.05). The 1-year cumulative re-bleeding rates were 41.5% in the shunt group and 19.5% in the shunt combined with embolization group (x2 = 6.320, P = 0.012). The differences of 1-year cumulative rates of shunt patency and mortality between the two groups were not significant (P is more than 0.05).

CONCLUSIONSTIPS combined with embolization of gastric coronary veins could reduce significantly the rate of rebleeding in 1 year after the shunt placement as compared with TIPS alone.

Adult ; Aged ; Embolization, Therapeutic ; Esophageal and Gastric Varices ; surgery ; Female ; Gastrointestinal Hemorrhage ; surgery ; Humans ; Male ; Middle Aged ; Portasystemic Shunt, Transjugular Intrahepatic ; Stomach ; blood supply

A slight halophilic denitrifying bacteria(designated GYL)was screened from the activated sludge which was used to treat high-salinity wastewater.According to the results of morphological observation,physiological and biochemical test,sequence analysis of the 16S rDNA,strain GYL was identified as Halo-monas sp..This strain could survive at 10% salinity and the optimal salinity range for growth was 2%~7%.The suitable pH value for growth was 7.5~8.5 and sucrose was the most effective carbon source.The nitro-gen removal efficiency exceeded 80% when the temperature ranged from 25?C to 30?C.Meanwhile hetero-trophic nitrification characteristics of this strain were measured.Results showed that this strain was able to realize SND and ammonia removal rate was 98.3%.It showed that this strain could perform the whole proc-ess of bacteria denitrification independently.

Objective To investigate the impact of miR-200c overexpression on colon cancer cell proliferation ability and the related mechanism.Methods MicroRNAs which may combined with the transcription factor AP-2α were screened and forecasted by the bioinformatics database,while its eukaryotic expression plasmids and specific inhibitor were synthesized.Plasmids PEZX-miR-200c,PEZX-NC,pmirGLO-AP-2α3'UTR,pmir-GLO and the specific inhibitors miR-67-inhibtor,miR-200c-inhibitor were transfected in vitro into colon cancer HCT-116 and SW480 cells and the HEK293T cell by Lipofectamine2000.The expression of AP-2α mRNA and protein in colon cancer cells was analyzed by qRT-PCR,Westem blot and immunocytochemical staining.CCK-8 assay and flow cytometry were adopted to observe the effect of miR-200c on colon cancer cells proliferation and apoptosis.Dual-Luciferase assay experiments were performed to observe the relative luciferase activity induced by miR-200c.Results The proliferation activity was significantly decreased in anti-miR-200c/SW480 group,while in PEZX-miR-200c/HCT-116 group,it was higher than that in PEZX-NC/HCT-116 group.The apoptosis ability was significantly increased in anti-miR-200c/SW480 group [(78±0.7) ％ vs (66±1.1) ％,P ＜ 0.05].The expression of AP-2o both in mRNA and protein levels was decreased in PEZX-miR-200c/HCT-116 group,while the protein level was increased in Anti-miR-200c/SW480 group.The relative luciferase activity inhibited by miR-200c was decreased in HEK-293T cells transfected with PEZX-miR-200c and pmir-GLO-AP-2α3' UTR (0.51±0.09 vs 0.98±0.04,P ＜ 0.01).Conclusion MicroRNA-200c could promote cell proliferation ability by targeting transcriptional factor AP-2α in human colorectal cancer cells.

Objective To evaluate the diagnostic value of the metabolic parameters for differentiating focal autoimmune pancreatitis (F-AIP) and pancreatic cancer (PC) by dual time 18F-FDG PET/CT scan.Methods Ten F-AIP patients and 20 PC patients in Changhai Hospital from May 2011 to November 2014 were enrolled in this study.All the AIP patients were histological confirmed or diagnosed by clinical follow up.The PC patients were histological confirmed and gender-and age-matched with F-AIP patients.50％ SUVmax was set as the threshold to fine-tune the boundary of interest.The extracted parameters included SUV SUV metabolic tumor volume (MTV),total lesion glycolysis (TLG),target-to-background ratio (TBR) and the retention indexes(RI) of all the parameters above.The PET/CT imaging features were also observed.Results The high metabolic lesions were observed in both F-AIP patients and PC patients.There were 6 F-AIP patients whose lesion was located in pancreas head,4 F-AIP patients whose lesion was located in pancreas body and tail.There were 12 PC patients whose lesion was located in pancreas head,8 PC patients whose lesion was located in pancreas body and tail.In F-AIP patients,2 cases had dilated pancreatic duct,6 had dilated biliary duct,8 had increased metabolism in mediastinal lymph node and 2 had abdominal lymphadenopathy,which were 8,5,5 and 14 cases in PC patients.The positive rate of mdeiastinal lymphadenopathy in F-AIP patients was statistically higher than that in PC patients,while the positivity rate of abdominal lymphadenopathy in AIP patients was lower than that in PC patients.The difference was statistically significant (both P ＜ 0.05).There were no statistical differences on the positivity rate of the dilated pancreatic duct,intra-and extra-hepatic bile duct between two groups.SUVmax,SUVmean and MTV in F-AIP were 5.37 ± 0.88,3.48 ± 0.66,21.79 ±15.60 in early stage and 6.45 ±1.51,4.23 ± 1.10,19.36 ± 14.63 in delayed stage,and those in PC were 8.31 ±3.08,5.41±1.95,9.26±8.35 in early stage,and 9.75±3.86,6.36±2.56,9.09±10.71 in delayed stage.SUVmax and SUVmean in F-AIP were lower than those in PC,whereas MTV were larger in F-AIP than that in PC.ROC curves for SUVmax,SUVmean and MTV were made.The AUC of SUV was the highest at 0.85,the cut-off value was 4.45,the corresponding sensitivity was 65％ and the specificity was 90％.TLG,TBR and RI of all the parameters were not statistically different in F-AIP and PC.Conclusions The 18F-FDG PET/CT metabolic parameters,such as SUVmax,SUVmean,MTV,could be of special diagnostic significance in discriminating F-AIP from PC.