3.Isolation of thermophilic bacteria Thermus sp. YBJ-1 and cloning of amylase gene.
Chinese Journal of Biotechnology 2004;20(3):434-436
Thermophilic bacteria strain YBJ-1 was isolated from hot spring samples collected from Yangbajing, Tibet. The 16sr DNA sequence of YBJ-1 (1511bp in length) shares 98% identity with that of Thermus scotoductus strain ITI-252T. The full-length ORF of amylase gene of YBJ-1 (amyT) was amplified by PCR technique and cloned into T-vector. The complete sequence of amyT is 1767bp in length, coding for 588 amino acids. The deduced amino acids share 99% similarity with alpha-cyclodextrinse of Bacillus sterothermophilus, 96% with maltogenic amylse of Thermus. sp IM6501, and 81% with neopullulanase of Bacillus sterothermophlus.
Amylases
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genetics
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Bacterial Proteins
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genetics
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Cloning, Molecular
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Culture Media
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Open Reading Frames
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genetics
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Thermus
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enzymology
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genetics
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isolation & purification
4.Characterization and gene cloning of the endoglucanase from Pseudoalteromonas sp. DY3 strain.
Chinese Journal of Biotechnology 2004;20(2):233-237
A bacteria strain DY3 with high endoglucanse activity was isolated from deep sea sediment sample ES0109. The 16S rDNA sequence of DY3 exhibits identity of 99% with those of the same genus bacteria Pseudoalteromonas citrea and Pseudoalteromonas elyakovii . The celX gene of DY3 obtained by PCR method is 1479bp in length and encodes a protein of 492 amino acids. The protein encoded by celX gene exhibits 95% sequence identity with endoglucanase CelG from Pseudoalteromonas haloplanktis. There are two modules in the deduced amino acids sequence, a catalytic domain of glycosyl hydrolases family 5 at the N terminal and a carbohydrate binding domain at the C terminal which was linked to catalytic domain by a short linker. The optimal temperature of CelX is 40 degrees C and the optimal pH was between 6 and 7.
Amino Acid Sequence
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Bacterial Proteins
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genetics
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Cellulase
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genetics
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Cloning, Molecular
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Gene Expression Regulation, Enzymologic
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Molecular Sequence Data
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Pseudoalteromonas
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enzymology
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genetics
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Sequence Alignment
5.Selective thoracic fusion in the scoliosis associated with syringomyelia.
Ke-Yi YU ; Jian-Xiong SHEN ; Gui-Xing QIU ; Jian-Guo ZHANG ; Yi-Peng WANG ; Yu ZHAO ; Bin YU
Chinese Journal of Surgery 2011;49(7):627-630
OBJECTIVETo evaluate the surgical results of selective thoracic fusion (STF) for scoliosis associated with syringomyelia.
METHODSFrom January 2001 to January 2009, 93 cases of scoliosis associated with syringomyelia were retrospectively reviewed. There were 11 cases who underwent STF and were followed up more than 2 years, which included 8 female and 3 male, the mean age was 14.9 years (9 - 21 years). Curve type, coronal and sagittal Cobb angle, apical vertebral rotation apical vertebral translation, flexibility, trunk shift were recorded and analyzed.
RESULTSThere were 9 double curves and 2 triple curves, the Lenke type of thoracolumbar/lumbar curve included Lenke A in 2 cases, Lenke B in 7 cases and Lenke C in 2 cases. The average coronal Cobb angle of thoracic curve before and after surgery were 62.6° and 19.0° respectively, and the average correction rate was 69.6%. The average coronal Cobb angle of thoracolumbar/lumbar curve before and after surgery were 36.1° and 11.6° respectively, and the average spontaneous correction rate was 67.9%. The followed up time ranged from 24 to 48 months (mean 29.5 months), the average loss of correction rate was 6.8%. Only one trunk decompensation was noted at final follow-up. Pedicle screw nut loosening occurred in one patient and this patient underwent revision surgery, no neurological complication was noted at final follow-up.
CONCLUSIONSSTF could be safely performed in scoliosis associated with syringomyelia. Thoracolumbar/lumbar curve in these patients has similar spontaneous correction ability compared with idiopathic scoliosis patients. The satisfactory result could be achieved according to the STF criteria for IS.
Adolescent ; Child ; Female ; Humans ; Male ; Retrospective Studies ; Scoliosis ; complications ; surgery ; Spinal Fusion ; methods ; Syringomyelia ; complications ; surgery ; Thoracic Vertebrae ; surgery ; Young Adult
6.Transcriptome profiling and analysis of Panax japonicus var. major.
Shao-peng ZHANG ; Jian JIN ; Bing-xiong HU ; Ya-yun WU ; Qi YAN ; Wan-yong ZENG ; Yong-lian ZHENG ; Zhang XI-FENG ; Ping CHEN
China Journal of Chinese Materia Medica 2015;40(11):2084-2089
The rhizome of Panax japonicus var. major have been used as the natural medicinal agent by Chinese traditional doctors for more than thousand years. Most of the therapeutic effects of P. japonicus var. major had been reported due to the presence of tetracyclic or pentacyclic triterpene saponins. In this study, Illumina pair-end RNA-sequencing and de novo splicing were done in order to understand the pathway of triterpenoid saponins in this species. The valid reads data of 15. 6 Gb were obtained. The 62 240 unigenes were finally obtained by de novo splicing. After annotation, we discovered 19 unigenes involved in ginsenoside backbone biosynthesis. Additionally, 69 unigenes and 18 unigenes were predicted to have potential function of cytochrome P450 and UDP-glycosyltransferase based on the annotation results, which may encode enzymes responsible for ginsenoside backbone modification. This study provides global expressed datas for P. japonicus var. major, which will contribute significantly to further genome-wide research and analysis for this species.
Gene Expression Profiling
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Panax
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genetics
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Saponins
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biosynthesis
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Sequence Analysis, RNA
7.The family investigation of a weak D type 15 donor.
Wen XIONG ; Jian-jiang QIN ; Yan LIU ; Yi-yan ZHOU ; Chao-peng SHAO
Chinese Journal of Medical Genetics 2007;24(1):35-37
OBJECTIVETo study the genetic feature of weak D type 15 allele (RHD845A) in a Chinese family.
METHODSRh D, C, c, E and e phenotypes of 4 members in a weak D type 15 family were tested by serological and polymerase chain reaction (PCR), D antigen was proven by indirect antiglobulin test. A pair of primers specific for RHD845A were designed, and a sequence specific primer-PCR (PCR-SSP) method was established to detect RHD845A allele in all family members. Subsequently the dual-tube PCR method was used to determine the RHD zygosity of 4 members.
RESULTSThe RHD845A allele existed in all 4 family members and the RHD zygosity test showed that all members were RHD +/RHD + homozygous. The parents and nephew possessed one normal RHD gene as RHD845A allele carriers, which caused RhD positive. The proband and his old-sister took two RHD845A alleles, which caused weak D phenotype.
CONCLUSIONThe proband is the weak D type 15 allele homozygous. The weak D type 15 gene is an ancestral allele, but not a mutation.
Adult ; Alleles ; Family Health ; Female ; Genotype ; Homozygote ; Humans ; Male ; Pedigree ; Point Mutation ; Polymerase Chain Reaction ; Rh-Hr Blood-Group System ; genetics
8.Effect of continuous passive motion on basic fibroblast growth factor expression during tendon-bone repair after surgical repair of acute rupture of the supraspinatus tendon in rabbits.
Sen LI ; Shao-xiong MIN ; Hui ZHANG ; Guo-jian FU ; Peng-cheng WANG ; An-min JIN
Journal of Southern Medical University 2010;30(5):1020-1023
OBJECTIVETo study the effect of continuous passive motion (CPM) on basic fibroblast growth factor (b-FGF) expression during tendon-bone repair in rabbits and explore the role of stress in the postoperative repair after acute rotator cuff injury.
METHODSSixteen rabbits randomized into CPM group (n=8) and non-CPM group (n=8) were subjected to surgically induced acute rupture of the supraspinatus tendon and subsequent surgical repair, with another two rabbits serving as the control. Two weeks after the operation, the rabbits in CPM group underwent CPM training, and those in non-CPM group were normally fed only. At 2, 4, 6, and 8 weeks after the operation, 2 rabbits from each group were sacrificed and the tissue samples were obtained for detecting the changes in b-FGF expression.
RESULTSTwo weeks after the operation, b-FGF expression was detected in both groups, and the CPM group showed slightly higher and more diffusive expression. At 4 weeks, b-FGF expression was significantly higher and distributed over a greater area in CPM group and in the non-CPM group. A large number of fibroblasts positive for b-FGF expression were identified in CPM group, aligning in parallel with the tendon membrane. At 6 weeks, b-FGF in the CPM group showed no obvious changes but that in the non-CPM group became lightened. At 8 weeks, b-FGF expression was reduced in both groups, which was more obvious in the non-CPM group.
CONCLUSIONCPM can promote b-FGF expression to enhance type III collagen synthesis at the tendon-bone interface in early stage of tendon-bone repair following acute rupture of supraspinatus tendon in rabbits, thereby contributing to tendon-bone recovery after rotator cuff injury.
Animals ; Bone Remodeling ; drug effects ; physiology ; Fibroblast Growth Factor 2 ; metabolism ; Male ; Postoperative Period ; Rabbits ; Random Allocation ; Range of Motion, Articular ; Rupture ; surgery ; Tendon Injuries ; rehabilitation ; surgery ; Tendons ; metabolism ; Wound Healing ; physiology
9.Clinical analysis of 33 cases of acute recurrent pancreatitis
Liangshun XIONG ; Jingyi WU ; Jian FEI ; Enqiang MAO ; Ruoqing LEI ; Jiancheng WANG ; Sheng CHEN ; Weize WU ; Tianquan HAN ; Chenghong PENG ; Yaoqing TANG ; Shengdao ZHANG
Chinese Journal of Hepatobiliary Surgery 2010;16(3):164-166
Objective To investigate the etiology of acute recurrent pancreatitis (ACP) and de-termine how to further enhance its level of treatment.Methods The clinical data of 33 patients with ACP treated in Ruijin Hospital from 2003 to 2007 were retrospectively analyzed.Results Of the 33 patients with an average age of 55 (22-86), 18 (55%) were male and 15 (45%) female.ACP occurred once in 26 patients, twice in 4 and 3 times in 3.The disease appeared whithin 1 year in 29 patients, 1-2 years in 2, 2-3 years in 1 and 3 years in 1 after being dischared from hospital.For its etiology, it was of biliary origin in 29 patients, hyperlipidemia in 1, pancreatic tumor in 1 and unknow reasons in 2.Twenty-four patients were treated with operation or endoscopy.Two patients died and the mortali-ty was 9.1%.Conclusion ACP is mainly due to biliary origin in China.Operative intervention at an appropriate opportunity can effectively reduce the recurrence of biliary-origin pancreatitis.
10.OATP1B1 in drug-drug interactions between traditional Chinese medicine Danshensu and rosuvastatin.
Jinhua WEN ; Xiaohua WEI ; Xiaohua CHENG ; Rong ZUO ; Hongwei PENG ; Yanni Lü ; Jian ZHOU ; Xuelian ZHENG ; Jun CAI ; Yuqing XIONG ; Li CAO
Acta Pharmaceutica Sinica 2016;51(1):75-9
The study was designed to explore the drug-drug interactions mechanisms mediated by OATP1B1 between traditional Chinese medicine Danshensu and rosuvastatin. First, the changes of rosuvastatin pharmacokinetics were investigated in presence of Danshensu in rats. Then, the primary rat hepatocytes model was established to explore the effects of Danshensu on the uptake of rosuvastatin by hepatocytes. Finally, HEK293T cells with overexpression of OATP1B1*a and OATP1B1*5 were established using a lentiviral delivery system to explore the effects of Danshensu on the uptake of rosuvastatin. Rosuvastatin pharmacokinetic parameters of C(max0, AUCO(0-t), AUC(0-∞) were increased about 123%, 194% and 195%, by Danshensu in rats, while the CL z/F value was decreased by 60%. Uptake of rosuvastatin in the primary rat hepatocytes was decreased by 3.13%, 41.15% and 74.62%, respectively in the presence of 20, 40 and 80 μmol x L(-1) Danshensu. The IC50 parameters was (53.04 ± 2.43) μmol x L(-1). The inhibitory effect of Danshensu on OATP1B1 mediated transport of rosuvastatin was related to the OATP1B1 gene type. In OATP1B1*5-HEK293T mutant cells, transport of rosuvastatin were reduced by (39.11 ± 4.94)% and (63.61 ± 3.94)%, respectively, by Danshensu at 1 and 10 μmol x L(-1). While transport of rosuvastatin was reduced by (8.22 ± 2.40)% and (11.56 ± 3.04)% and in OATP1B1*1a cells, respectively. Danshensu significantly altered the pharmacokinetics of rosuvastatin in rats, which was related to competitive inhibition of transport by OATPJBI. Danshensu exhibited a significant activity in the inhibition of rosuvastatin transport by OATP1B1*5-HEK293T, but not by OATP1B1*1a, suggesting a dependence on OATP1B1 sequence.