2.Cloning and prokaryotic expression analysis of squalene synthase 2 (SQS2) from Salvia miltiorrhiza f. alba.
Qi-xian RONG ; Dan JIANG ; Liang-ping ZHA ; Ye SHEN ; Yan ZHANG ; Lu-qi HUANG
China Journal of Chinese Materia Medica 2015;40(7):1259-1265
According to the designed specific primers of gene fragment based on the Salvia miltiorrhiza transcriptome data, a full-length cDNA sequence of SQS2 from S. miltiorrhiza f. alba was cloned by the method of reverse transcription polymerase chain reaction (RT-PCR). The SmSQS2 cDNA sequence was obtained, this sequence is named SmSQS2 and its GenBank registration number is KM244731. The full length of SmSQS2 cDNA was 1245 bp, encoding 414 amino acids including 5'UTR 115 bp and 3'UTR 237 bp. Sequence alignment and phylogenetic analysis demonstrated that SmSQS2 had relative close relationship to the SQS2 of S. miltiorrhiza. The induction of E. coli [pET28-SQS2] in different temperature, induction time, IPTG concentrations and density of inducing host bacterium (A600) were performed, Shaking the culture at 30 degrees C until the A600 is approximately 0.6 and add IPTG to final concentration of 0.2 mmol x L(-1), and then the optimal expression of SmSQS2 recombinant protein were accumulated after the induction time of 20 h. The research provided important base for the study of sterol and terpene biosynthesis of SQS2 in S. miltiorrhiza f. alba.
Cloning, Molecular
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Farnesyl-Diphosphate Farnesyltransferase
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chemistry
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genetics
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metabolism
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Models, Molecular
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Molecular Sequence Data
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Phylogeny
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Plant Proteins
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chemistry
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genetics
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metabolism
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Salvia miltiorrhiza
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chemistry
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classification
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enzymology
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genetics
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Sequence Alignment
3.Research Progress About Some Problems on Cordyceps militaris
Yan-Feng HAN ; Jian-Dong LIANG ; Wen DU ; Zong-Qi LIANG ;
Microbiology 2008;0(09):-
Cordyceps militaris is of significant economic value in medical care and food exploitation because of its many physiological activities. This paper reviews(1) the taxonomic position of its anamorph,(2) interesting culture ways,(3) strain degeneration and genetic variability,and(4) research progress in bioactive compounds and pharmacological functions.
4.Nephrotoxicity study of Aristolochia fangchi in rats by metabonomics.
Qi LIANG ; Cheng NI ; Ming XIE ; Qi ZHANG ; Yanxia ZHANG ; Xianzhong YAN ; Meijuan YANG ; Shuangqing PENG ; Yuzhong ZHANG
Journal of Integrative Medicine 2009;7(8):746-52
To study the changes of metabolites in rat urine after treatment of Aristolochia fangchi decoction by metabonomic method.
5.Factors associated with prevalence, awareness, treatment and control of hypertension among adults in Dagang Oil Field:A cross-sectional survey
Wenwei QI ; Hui JIA ; Yuanyuan WANG ; Yan CHEN ; Yingzi LIANG ; Lifeng LI ; Jian WEI ; Guangping LI
Chinese Journal of Health Management 2015;(1):55-60
Objective To investigate factors associated with prevalence, awareness, treatment and control of hypertension in Dagang Oil Field, and to provide the scientific evidence for the intervention measurements. Methods A cross-sectional, population-based survey was conducted in Dagang Oil Field from 2008 to 2010. Totally, 16 553 adults, 8 174 men (49.38%) and 8 379 women (50.62%), completed all of the questionnaires, measurement examination and blood specimen collection. Results The prevalence of hypertension was 27.30%. Multivariate logistic regression analysis showed that male, old age, overweight or obesity, family history of hypertension, marriage, education attainment, snoring, high heart rate, fasting blood glucose, serum total protein, and hyperuricemia were associated with hypertension. The awareness, treatment, control rates among all hypertensive subjects, and control rate among treated hypertensive participants were 52.65%, 40.07%, 15.51%, and 38.78%, respectively. Awareness was more common in those who were 45-64 years old , high medical costs, family history of hypertension, and smoking currently. Odds ratio (OR) (95% CI) of awareness was 5.24 (2.78 to 9.88), 24.11 (11.91 to 48.84) and 27.46(13.92 to 54.18) for medical costs 1-59.9 yuan/month, 60-199.9 yuan/month, and≥200 yuan/month, respectively. In contrast, exercising 2-4 times/week and longer sleep duration were associated with lower awareness. Medication treatment was more common in those who had old age, high medical costs, hypertension family history, coronary heart disease, prefer vegetable, while less common in those who exercise occasionally, sleep longer and drink alcohol currently. OR (95%CI) of treatment was 10.89 (3.76 to 31.56), 54.07 (19.20-152.24), and 87.31 (31.54 to 241.75) for medical costs 1-59.9 yuan/month, 60-199.9 yuan/month, and ≥200 yuan/month, respectively. Controlled hypertension was more common in those who had old age, high medical costs, exercise 2-4 times/week, coronary heart disease, prefer crude fibre diet, but less common in those who drink alcohol currently, have higher heart rate, and higher education attainment. OR (95%CI) of controlled hypertension was 13.59 (6.50-28.44), 14.67 (7.01-30.68), and 17.43 (8.34-36.39) for medical costs 1-59.9 yuan/m, 60-199.9 yuan/m, and≥200 yuan/m, respectively. Conclusion Despite high rate of awareness, treatment and control of hypertension still need to be strengthened.
6.MRI study of the infantile developmental dislocation of the hip
Liang WANG ; Mao SHENG ; Wanliang GUO ; Qi WANG ; Yan REN ; Yunfang ZHEN ; Yi YANG ; Junkang SHEN
Journal of Practical Radiology 2014;(4):649-652
Objective To investigate the diagnostic value of MRI in the infantile developmental dislocation of the hip.Methods Forty-five cases of preoperative hip MRI data of DDH (patient group)and 40 cases of normal hip (control group)MRI data were retrospectively analyzed.Three types were classified according to the standard of Dunn.Then the bony acetabular index (BAI),cartilaginous acetabular index (CAI)were measured on coronal MRI and all the intra-articular structures (articular cartilage,labrum, teres ligament and iliopsoas)in the acetabulum that impeded the reduction of the femoral head were observed.Results There were normal hips (80 hips)and dislocated hips (73 hips).In dislocated hips typeⅠ (30 hips),typeⅡ (20 hips),and typeⅢ (23 hips) were found.The values of BAI were (26.13±2.19)°and (35.87±4.42)°,CAI (13.84±1.07)°and (21.92±4.70)°for patient group and control group.The values of BAI were (31.80±1.74)°,(37.80±3.40)°and (39.17±4.11)°,CAI (17.20±2.68)°,(20.91±2.02)°and (27.50±2.78)°for the typesⅠ,Ⅱ,Ⅲ respectively.Differences of BAI and CAI between patient group and control group were showed (t=-11.048 and -9.188,P =0.000).Statistically significants were found among the typeⅠ,Ⅱ,Ⅲ(F=22.159 and 47.241,P =0.000).There were linear correlations for patient group ,control group,and the typeⅠ,Ⅱ,Ⅲ (r= 0.964,0.844,0.953,0.931 and 0.870,P =0.000).Some pathological changes such as varus labrum,thick teres ligament,iliopsoas spasm,articular cartilage hyperplasia,fat hyperplasia,joint effusion were observed in the type Ⅱand Ⅲ.Conclusion MRI is not only used for measuring the values of BAI and CAI,and a linear correlation among all kinds of DDH,but also some factors hindering the reduction of the femoral head can be found clearly.It can provide an important reference basis for the diagnosis and treatment of DDH.
7.Construction and effect of Neuropilins-2 eukaryotic expression vector for RNA interference
Qi ZHOU ; Houjie LIANG ; Xiaochu YAN ; Qiuping PENG ; Jinming ZHOU ; Feng WU ; Daping ZHONG ; Zhiheng BIAN
Journal of Third Military Medical University 2003;0(08):-
Objective To construct Neuropilins-2 eukaryotic expression vector for RNA interference.Methods Recombinant targeting on gene NRP2 was designed and established with plasmid pGenSil-1 based on NRP2 cDNA equences of Genomes.Two pairs of oligonucleotides were synthesized according to the Tuschl and inserted into plasmid pGenSil-l to generate siRNA eukaryotic expression vector,DH5? strains were transformed,plasmid were extracted,and recombinant vectors were identified by the restriction map and the sequence analysis.The recombinant plasmid(pGenSil-NRP2) was transfected into the cultured LOVO cells.At 48 h after transfection,the whole cell protein was extracted,and the protein level was detected by Western blotting with mouse-anti-human NRP2 monoclonal antibody.Results Recombinant plasmids were completely coincided with the designs by the restriction map and the sequence analysis.pGenSil-NRP2 expression vector into LOVO cells down-regulated the protein level of NRP2 at 48 h after transfection.The recombinant eukaryotic expression vector were constructed successfully.Conclusion siRNA recombinant can be constructed successfully by RNAi technique for inhibiting NRP2 expression.
8.The cAMP-mediated protein kinase signal transduction pathway is involved in the pyrogenic effect of CRH in rats
Huadong WANG ; Yanping WANG ; Yang QU ; Renbin QI ; Daxiang LU ; Chujie LI ; Liang YAN
Chinese Journal of Pathophysiology 2001;17(8):785-
The primary action of corticotropin releasing hormone (CRH) is stimulation of the synthesis and release of adrenocorticotropic hormone and β-endorphin from the pituitary in response to stress. In addition, a number of studies indicate that CRH exerts other physiological actions within the central nervous system which are independent of the pituitary. These include increased body temperature and thermogenesis. However, the intracellular mechanism responsible for pyrogenic action of CRH is still unclear. The purpose of these studies was to determine whether or not cAMP was involved in the pyrogenic action of CRH in the rat. Intracerebroventricular (icv) microinjection of CRH (2.5 μg, 5.0 μg, 10 μg) caused increases in colonic temperature and hypothalamus cAMP level in conscious rats. The pyrogenic effects of CRH were abolished or markedly inhibited by prior injection (icv) of an adenylate cyclase inhibitor, 2,,3,-dideoxyadenosine (DDA, 30 μg) or an inhibitor of cAMP-dependent protein kinase, adenosine-3,,5,-(cyclic) monophosphorothionate (Rp-cAMPs, 15 μg). This is the first report demonstrating the pyrogenic effcet of centrally administration of CRH on the rat via the cAMP-mediated protein kinase signal transduction pathway.
9.Treatment of thoracolumbar spinal tuberculosis with single posterior debridement, bone grafting, internal fixation and local chemotherapy.
Ying-Qi YAN ; Man-Li XIA ; Jia-Ping DAI ; Sui-Liang GONG ; Gang CHEN ; Xiao ZHOU
China Journal of Orthopaedics and Traumatology 2015;28(4):323-326
OBJECTIVETo evaluate the clinical effects of single posterior debridement, bone grafting, internal fixation and local chemotherapy in treating thoracolumbar spinal tuberculosis.
METHODSFrom February 2009 to September 2012,11 patients with thoracolumbar spinal tuberculosis were treated by single posterior debridement, bone grafting, internal fixation and local chemotherapy. There were 7 males and 4 females, aged from 27 to 65 years old with an average of 53.7 years. The courses of disease was from 3 months to 2 years with the mean of 9 months. According to ASIA standard of spinal cord injury, 3 cases were grade C and 8 cases D. After treatment, clinical effects were evaluated by ASIA grade, visual analogue score (VAS) and Oswestry Disability Index (ODI); kyphosis Cobb angle change was observed by X-rays.
RESULTSEleven patients were followed up from 12 to 29 months with an average of 18 months. ASIA grade of spinal cord injury, 3 patients with grade C improved to grade D in 2 cases and grade E in 1 case 8 patients with grade D improved to grade E in 7 cases and unchanged in 1 case. VAS decreased from preoperative 6.10 ± 1.30 to 1.70 ± 0.80 at 3 d after operation (P < 0.05). ODI improved from preoperative (68.36 ± 10.41)% to (14.55 ± 8.99)% (P < 0.05) at 3 d after operation. Kyphotic Cobb angle was corrected from preoperative (22.64 ± 4.84)° to (4.27 ± 1.49)° (P < 0.05) on the 3rd day after operation, and angle loss was mild at final follow-up, there was no significant difference between postoperative at 3 d and final follow-up.
CONCLUSIONSingle posterior debridement, bone grafting, internal fixation and local chemotherapy for the treatment of thoracolumbar spinal tuberculosis can effectively remove the lesion, improve nerve function and correct deformity, has advantage of single incision, little trauma, and low recurrence rate. But it still need long-term and systemic treatment with anti-TB drugs.
Adult ; Aged ; Bone Transplantation ; Debridement ; Female ; Humans ; Internal Fixators ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Thoracic Vertebrae ; surgery ; Tuberculosis, Spinal ; therapy
10.Purification, Biochemical Properties, and Activities of a Novel Factor X Activator (F V e-1 ) from Daboia Russelli Siamensis ( Myanmar ) Venom
Xi LIN ; Shubo XIN ; Jiezhen QI ; Xiuxia LIANG ; Jiashu CHEN ; Pengxin QIU ; Guangmei YAN
Journal of Sun Yat-sen University(Medical Sciences) 2012;33(2):141-148
[Objective] To purify and characterize a novel factor X activator,Fve-1 from Daboia russelli siamensis (Myanmar) venom.[ Methods]F V e-1 was purified by ion-exchange chromatography and gel filtration.The hemostatic activity of F V e-1 was determined based on chromogenic substrates.The fibrinogen-clotting activity of F V e-1 was also determined.Thermal stability, pH stability,enzyme activity,and inhibition of F V e- 1 were determined by its remaining procoagulant activity.N-treminal sequence was determined by the method of automated Edman degradation.[ Results ]F V e-1 was achieved by chromatography with a molecular weight of 13,808 and an isoelectric point of 4.6. The hemostatic activity of 0.5 mg Fve-1 was equal to that of 1.5625 u thrombin or that of 54.93 ng RVV X. F V e-1 primarily activated F X, but did not affect on prothrombin and fibrinogen. The suitable pH and temperature range of F V e-1 was 6.5-7.5 and 25-60 ℃,respectively.The activity of F V e-1 was enhanced by Ca2+ and inhibited by EDTA and DTT.The N-terminal sequence of F V e-1 was NH2-N-L-Y-Q-F-G-E-M-I-N.[Conclusion] F V e-1 is a factor X-activating enzyme,which could activate FX to FX a,but have minimal effect on prothrombin and fibrinogen.