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.
6.Variety of corneal endothelial cell in glaucoma by confocal microscope
Hong-Liang, GAO ; Zhi-Qiang, GUO ; Hong-Yu, FAN ; Li-Juan, XU ; Yan-Hua, QI
International Eye Science 2014;(10):1831-1833
ATM: To define the causes of corneal endothelial cell damage, to investigate the preventive methods, and to observe the variety of corneal endothelial cell in glaucoma using confocal microscope.
METHODS: Totally, 143 eyes of 97 patients with different types of glaucoma, and matched normal people were 20 cases, all 40 eyes. The cell density, cell area and cell variable coefficient were measured used confocal microscope. These indicatives of every kind of glaucoma were compared.
RESULTS: The corneal endothelial cell density of normal group was 2 893. 88±255. 026/mm2 , the group of acute angle-closure glaucoma ( AACG ) was 1 674. 11±683.95/mm2 , and the group of open angle glaucoma (OAG) was 2687. 22±391. 87/mm2, the group of chronic angle-closure glaucoma (CACG) was 2706. 97±351. 27/mm2. In all index the average cell density of corneal endothelial and the average area have statistical significance ( F =62.950, 8. 795;P=0. 000), especially the group of AACG.CONCLUSION: The index of corneal endothelial cell in AACG is lower than that of normal. All index in OAG and CACG is difference with that of normal, but the difference has no statistical significance. And the dominant factor of damaged corneal endothelial is the time of intraocular hypertension.
7.Effects of iodine deficient and hypothyroid on protein expression of extracellular signal-regulate kinase (ERK1/2) in the cerebellum of rats
Jin-yan, LI ; Jing, DONG ; Yi, WANG ; Jian, GONG ; Liang, SHAN ; Ming-qi, GAO ; Jie, CHEN
Chinese Journal of Endemiology 2009;28(3):239-243
Objective To study the effects of iodine deficiency and hypothyroidism on protein expression of extracellular signal-regulate kinase(ERK1/2) in the cerebellum of rots. Methods Twenty-eight healthy Wistar rots, female, 60 days old, were randomly divided according to their body weight into control group, iodine deficient group and hypothyroidism groups. Hypothyroidism groups in accordance with drinking water containing propylthiouracil(PTU) were divided into doses of 5 mg/L and 15 mg/L groups, 7 rats in each group. Rats after pregnancy, iodine deficient rats were administered with iodine-deficient diet and hypothyroid rats were administered with PTU in drinking water. Pup's cerebellum in each group were weighed on day 14,21,28 and 42. Cerebellum tissue was observed for cerebellar morphology using silver staining and detected for ERK1/2 protein using immunohistochemistry on day 7,14,21,28 and 42. Results On day 14,21,28 and 42, cerebellum weight of pups from iodine-deficient[(0.0945±0.0233), (0.1347±0.0046), (0.1542±0.0094), (0.1949±0.0048)g]were significantly lighter than control[(0.1856±0.0123), (0.2049±0.0098), (0.2268±0.0065), (0.2606±0.0086)g, all P < 0.05]and hypothyroidism groups [for 5 mg/L group: (0.1741±0.0172), (0.1927±0.0103), (0.2181±0.0064), (0.2583±0.0054)g, all P<0.05; for 15 mg/L group: (0.1604±0.0083), (0.1682±0.0103), (0.1996±0.0073) and (0.2579±0.0067)g, all P< 0.05]the difference had statistical significance(F=6.325,8.870, 16.191 and 21.574, all P<0.05). Compared to the controls on day 7, iodine-deficient group didn't have clear layers; on day 21, disappearance of external granule cells from iodine-deficient and 15 mg/L groups was delayed, still two or three layer external granule cells remained; on day 28 and 42, molecular layer from 5, 15 mg/L and iodine-deficient groups and became thinner. Immunohistochemistry showed that on day 7, there was no statistical difference of integrated optical density average of ERK1/2, in all the groups(F=1.102, P>0.05); on day 14,21,28 and 42, integrated optical density average of ERK1/2 in iodine-deficient group[(7.3245±0.5070), (8.3606±1.0683), (9.1217±1.0402), (12.1587±0.7581), all P<0.01]and hypothyroidism groups [for 5 mg/L group: (11.4307±1.5200), (14.919±0.8497), (16.0082±1.1130), (15.7721±0.8293), all P< 0.01; for 15 mg/L group: (7.8538±0.9775), (11.2461±0.8138),(12.78±1.3783), (13.0871±1.1450), all P < 0.01]was significantly lower than those of controls [(16.2831±0.5143), (20.2653±0.9551), (22.7485±1.0267), (22.1725±0.9939), all P < 0.01], the difference having a statistical signifieance(F=16.131,13.543,26.953,41.583, all P<0.01). Conclusions Iodine deficiency and hypothyroidism during critical periods of brain development may change eerebellar morphology and down regulate the protein expression of ERK1/2, which may result in damage of cerebellum development.
8.Investigation and analysis of trace element levels in children with osteomalacia malformation from endemic fluorosis area in Shuicheng, Guizhou
Fu-cheng, LI ; Liang, QI ; Xu-zi, CONG ; Ying, YANG ; Shu-lin, YAN
Chinese Journal of Endemiology 2011;30(2):197-201
Objective To investigate the levels of trace elements such as fluorine(F), and aluminium (Al)etc. of osteomalacia malformation children and to make etiological diagnosis in reference with clinical manifestations.Methods Urine and occipitalia hairs of 14 diseased children(patient group) from endemic fluorosis area and 13 healthy children(control group) from non-endemic area were included in the study on November, 2008, and contents of 10 elements of fluorine(F), aluminum(Al), chromium(Cr), manganese(Mn), ferrum(Fe), cuprum(Cu), zinc(Zn), arsenic (As), selenium(Se), strontium(Sr), and barium(Ba) were tested. The data were analyzed with medical soft package PEMS 3.1. Results Urinary contents of F, Al, Mn, Cu, Sr, and Se(1.18 mg/L, 112.6 μg/L,6.62,29.86 mg/L, 177.5,4.23 ng/L) in patient group were significantly different from those in control group (0.48,47.1,2.04,16.61 mg/L, 55.17,15.52 ng/L, t = 4.592,2.486,4.850,2.210 2.078,2.912, all P< 0.05); Hair contents of Al, Mn, As, Sr, Ba, Fe, and Se in patient group(59.27,5.26,0.96,1.50,1.29,297.13,0.45 mg/kg)were significantly different from those of control group( 18.69,0.72,1.09,0.62,0.68,69.02,1.323 mg/kg, t = 4.583,6.318,3.309,2.704,5.606,6.294, all P < 0.05); in patient group, the correlation coefficients of urinary Fe to Al,Zn, As, and Se were all bigger tan 0.662(all P< 0.05), those of urinary Se to Mn, Ba, Cu, Zn, Sr, and As were all bigger than 0.694(all P< 0.05), those among urinary Mn, Sr, As, and Ba were bigger than 0.550(all P<0.05), those of hair Al to Mn, Cr, Fe, and Cu were bigger than 0.732(all P< 0.05), those of hair Ba to Mn,Cr, Fe, and Sr, and of hair Mn to Cr and Fe, and those between Cr and As, between Cu and Sr were all bigger than 0.686 (all P < 0.05). In control group, the correlation coefficients of urinary Cu to Zn, Se, and Ba, those of Zn to Se and Ba, and those of Cr to Mn and Ba were all bigger than 0.516(all P < 0.05), those of hair Al to Mn,Fe, Cu, As, and Se, and those of hair Se to Fe, Cu, and As, those of hair Fe to Mn, Cu, and As, those of hair Cu to Zn and As, and that between Zn and As were bigger than 0.739(all P < 0.05). The correlation coefficient of urinary F to Se in patient group(0.762) was significantly different from that in control group( - 0.469, u = 2.079,P < 0.05). Conclusions The burden of F and Al of osteomalacia malformation children in endemic fluorosis area of Shuicheng county is too high. The contents of multi-elements in urine and hairs and their correlation are coincident with high levels of Al and F and they cause network increase of multi-element content changes and their correlation. According to bone X-ray features combining with the living environment, the diagnosis of endemic Al-F fluorosis can be made. The biological significance of reducing urinary and hair Se levels and the correlations of F and Al need to be further studied.
9.lnhibition effect of AG490 on high glucose-induced epithelial-mesenchymal transition in human lens epithelial cells
Li, FENG ; Liang, ZHANG ; Xiao, HAN ; Bo, LU ; Xin-Ling, WANG ; Qi-Chang, YAN
International Eye Science 2014;(12):2122-2126
AlM:To study the relationship between JAK-STAT pathway and epithelial - mesenchymal transition in human lens epithelial cells. Meanwhile, the function of AG490 as a JAK inhibitor was also demonstrated in this article.
METHODS:Human lens epithelial cells SRA01/04 ( LECs ) were treated by low concentration of glucose (5. 5mmol/L). High concentration of glucose (30. 5mmol/L) was used to treat the cells in order to form the high glucose model. According to adding AG490 or not, cells were divided into the control group and the experimental group, appropriate concentration 10ü mol/L and 50ü mol/L of AG490 were chosen and acting time of 6, 12, 24, 48h were selected. Effect of AG490 on cell migration was measured by wound healing test. The expression of TGF-β1 , FN,α-SMA mRNA were examined by RT-PCR.
RESULTS:With the prolonged acting time ( 6, 12, 24 and 48h), cell activity increased in the HG group, as well as more expression of TGF-β1 , FN,α-SMA mRNA were detected compared to the LG group (P<0. 05). ln AG490 group, the cell migration activity and expression of TGF-β1 , FN,α-SMA mRNA decreased compared to the HG group (P<0. 05).
CONCLUSlON:JAK-STAT pathway takes part in high glucose-induced epithelial-mesenchymal transition in human lens epithelial cells. The mechanism is that it impacts the transcriptional expression of TGF- β1 and extracellular matrix. AG490, a JAK inhibitor, inhibits high glucose-induced epithelial-mesenchymal transition in human lens epithelial cells, And the inhibition enhances with the increasing concentration of AG490.
10.Clinical Evaluation of S-Adenosyl-L-Methionine in Treatment of Jaundice in 202 Newborn Infants
qi-liang, CUI ; hui, ZHANG ; hai-yan, LIU ; yuan-qing, LIN ; hui-yuan, TAN
Journal of Applied Clinical Pediatrics 2004;0(08):-
Objective To observe the effect of S-adenosyl-L-methionine(SAMe) in treatment of jaundice in newborns and its mechanism.Methods Two hundred and two newborn infants with jaundice were treated with SAMe,76 cases in control group treated with phototherapy and liver enzyme induction elixir;SAMe 30-60 mg/(kg?d) were added to 202 cases intravenously in treatment group.The total biliorubin(T-BILI),direct bilinrubin(D-BILI) and indirect bilinrubin(I-BILI) were dynamically detected.Results Six days after treatment,the skin jaundice index in treatment group decreased remarkably.T-BILI,D-BILI and I-BILI decreased significantly.The curing effectiveness was higher in treatment group than that in control group.The number of applicating blood products and albumin,and blood produets/albumin were decreased in treatment group than those in control group.In those who used glucose to dissolve the SAMe 2.68% had blood-vessel phlebitis.Conclusions SAMe can efficiently quicken the retrogression of jaundice in newborns.It can reduce the use of blood products.It is a reliable and safe drug to treat jaundice in newborns.