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.Chemical constituents from Crepis crocea.
Yuan-yuan LI ; Zhao-qi PENG ; Shi-lin HE ; Yan NI ; Xu-liang HAO
China Journal of Chinese Materia Medica 2015;40(19):3800-3804
Thirteen compounds were isolated from the ethyl acetate fraction of Crepis crocea by column chromatographies on silica gel, Sephadex LH-20 and semi-preparative HPLC. The structures were elucidated on the basis of spectral analysis as tectorone I (1), 8β- (2-methyl- 2-hydroxy-3-oxobutanoyloxy) -glucozaluzanin C (2), tectoroside (3), luteolin-7-O-glucoside (4), cosmosiin (5), esculetin (6), 3,4-dihydroxybenzaldehyde (7), trans-4-hydroxycinnamic acid (8), Caffeic acid (9), methyl p-hydroxyphenyllactate (10), ethylp- hydroxyphenyllactate (11), cis-3,4-dihydroxy-β-ionion (12). All the compounds, except for compounds 4 and 9, were isolated from this plant for the first time, and tectorone I (1) is a new natural product.
Crepis
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chemistry
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Drugs, Chinese Herbal
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chemistry
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isolation & purification
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Mass Spectrometry
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Molecular Structure
6.Effects of B7-1 on the cytoskeleton rearrangement in podocytes induced by angiotensin Ⅱ
Aimin DU ; Xinghua CHEN ; Qian YANG ; Qi YAN ; Wei LIANG ; Guohua DING
Chinese Journal of Nephrology 2017;33(2):126-131
Objective To observe the effect of costimulatory molecule B7-1 on cytoskeleton rearrangement in mouse podocytes induced by angiotensin Ⅱ (Ang Ⅱ),and to study the underlying molecular mechanism of B7-1 in the pathological changes of podocytes.Methods All cultivation of conditionally immortalized mouse podocytes (MPC) in vitro were divided into the following groups:normal control group,CTLA-4 group,Ang Ⅱ group (10-6 mmol/L 12 h,24 h;10-8 mmol/L 12 h,24 h) and CTLA-4 with Ang Ⅱ group.Transfect B7-1 RNA interference fragment (siRNA) to the mature podocytes,and then restimulated by Ang Ⅱ (10-6 mmol/L 12 h),the change of podocyte cytoskeleton after Ang Ⅱ stimulation were observed.The expression of B7-1 in each group was assayed by flow cytometry and Western blotting.The nephrin and p-nephrin protein levels in the four groups were also analyzed by Western blotting.At the same time,the podocyte cytoskeleton distribution as indicated by F-actin was observed by fluorescence microscopy.Results Flow cytometry and Western blotting showed that B7-1 was not expressed in the normal control group.Ang Ⅱ showed a concentration and time dependent induction of B7-1 expression in mouse podocytes (P < 0.05).Western blotting indicated that Ang Ⅱ induced B7-1 protein expression (P < 0.05).Expression of nephrin and p-nephrin was significantly down-regulated by Ang Ⅱ (P < 0.05).Compared with the normal control group,the expression of podocyte protein nephrin and p-nephrin in Ang Ⅱ stimulation group was significantly reduced (P < 0.05).Using FITC phalloidin fluorescence staining showed that CTLA-4+Ang Ⅱ stimulation group cytoskeleton rearrangement was improved significantly and F-actin recombinant score (mCFS) decreased compared with Ang Ⅱ group (P < 0.05),suggesting that Ang Ⅱ led to the disorder of the podocytes cytoskeleton and the destruction of the cytoskeleton of podocytes by Ang Ⅱ could be improved after B7-1 blocking.Compared with the Ang Ⅱ stimulation,transfection of B7-1 siRNA + Ang Ⅱ stimulation group improved F-actin cytoskeletal rearrangement,and mCFS also decreased significantly (P < 0.05),suggesting that transfection of B7-1 siRNA might improve the damage of Ang Ⅱ on podocytes cytoskeleton.Conclusions B7-1 participates in the process of cytoskeleton reconstruction and plays an important role in the pathological changes of podocytes.
7.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
8.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.
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.