1.Cloning and bioinformatic analysis and expression analysis of beta-glucuronidase in Scutellaria baicalensis.
Shuang-shuang GUO ; Lin CHENG ; Li-min YANG ; Mei HAN
China Journal of Chinese Materia Medica 2015;40(22):4370-4377
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
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Base Sequence
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Cloning, Molecular
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Computational Biology
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Glucuronidase
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chemistry
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genetics
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metabolism
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Molecular Sequence Data
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Open Reading Frames
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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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Protein Structure, Secondary
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Scutellaria baicalensis
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chemistry
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enzymology
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genetics
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Sequence Alignment
2.Study on acute inhalation toxicity of Luowei,a plant molluscacide
Ping YI ; Cheng PENG ; Shuang LI
Chinese Journal of Schistosomiasis Control 2016;(1):79-80
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.
3.THE EXPERIMENTAL EFFECTS OF FREEZING THE AREA OF FIRST PORTA HEPATIS IN PIGS
Jianping LIU ; Qingjia OU ; Shuang CHENG
Medical Journal of Chinese People's Liberation Army 1981;0(04):-
To investigate the effects of freezing the area of first porta hepatis which involves Glisson duct system,16 pigs were randomly divided into 2 groups. Group A:structures of the first porta hepatis were frozen by a dish-like cryoprobe (D=3.0cm) with temporary blockage of hepatic portal blood during freezing,the temperature at the end of 1,2,3minutes of cryotherapy was -50℃,-110℃,-131℃.Group B:sham operation,only with hepatic portal blockage for 3 minutes. Liver function tests, 99mTc-EHIDA scintiscanning,and pathological observation were used to evaluate the effects of the cryosurgery postoperatively.The results showed that the bile duct systems on first porta hepatis were severely destroyed by the cryosurgery,the cryoimpairment of common bile duct and/or common hepatic duct led to complications of progressive, irreversible, cryptic necrosis, fistula and pyogenic angiocholitis. The large freezing necrotic lesion in the porta hepatis might compress the portal vein. The perfect protection of biliary system should be accomplished in the cryosurgery of the first porta hepatis to avoid severe complications.
4.Advances in personalized treatment of small cell lung cancer
Shuang ZHANG ; Jingjing LIU ; Ying CHENG
Chinese Journal of Clinical Oncology 2017;44(12):571-576
Small cell lung cancer (SCLC) is a lethal malignancy characterized by rapid growth, early metastatic spread, and unfavorable survival outcomes. Optimizing treatment for patients with SCLC has been the focus for investigators. The emergence of precision medi-cine and personalized treatment brought significant breakthroughs into SCLC treatment and changed the therapeutic model. The de-velopment of molecular bioinformatics increased our understanding of complex molecular mechanisms of SCLC, and novel targets for personalized treatment have been developed. Clinical trials testing these targets are ongoing, which show the potential of personal-ized treatment for SCLC.
5.Establishment and application of a high-throughput drug screening model based on COL1A1 promoter for anti-liver fibrosis.
Shuang-Shuang ZHAO ; Ju-Xian WANG ; Yu-Cheng WANG ; Rong-Guang SHAO ; Hong-Wei HE
Acta Pharmaceutica Sinica 2015;50(2):169-173
For screening the potential drugs as anti-liver fibrosis candidates, we established a high- throughput drug screening cell model based on COL1A1 promoter. The activity of COL1A1 promoter and luciferase reporter gene can be elevated by TGF-β1, and inhibited by candidate drugs. We constructed a recombined plasmid with COL1A1 promoter and luciferase reporter gene pGL4.17, the activity of COL1A1 promoter was reflected by fluorescence intensity. COL1A1 promoter activity was detected by Dual-Luciferase Reporter Assay System, it came that the relative luciferase activity of COL1A1 promoter was 15.98 times higher than that of control group induced by TGF-β1, showing the recombined plasmid could be used in cell model. The recombined plasmid was transfected into human hepatic stellate cells LX2, detected the effect of potential drugs, and obtained a stable expression system through stable transfection and monoclonal cell culture. A sample which could reduce COL1A1 promoter activity signally by our cell model, decreased collagen I mRNA and protein expression detected by real-time RT-PCR and Western blotting. It indicates this novel cell model can be used in high-throughput drug screening of potential anti-liver fibrosis drugs.
Collagen Type I
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genetics
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Drug Evaluation, Preclinical
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methods
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Genes, Reporter
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Hepatic Stellate Cells
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High-Throughput Screening Assays
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Humans
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Liver Cirrhosis
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drug therapy
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Luciferases
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Plasmids
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Promoter Regions, Genetic
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RNA, Messenger
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Transfection
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Transforming Growth Factor beta1
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pharmacology
6.Effects of a new diet intervention for college teachers on their dietary behavior change
Shuang LIU ; Qiuli ZHAO ; Yanqiu WANG ; Jiangping MA ; Shanshan CHENG ; Shuang CANG
Chinese Journal of Practical Nursing 2016;32(32):2502-2506
Objective To construct the new diet intervention scheme of dyslipidemia of university teachers, discuss the effect of this plan on eating behavior change. Methods Choose two groups of college teachers in Harbin, who were hyperlipemia and had physical examinations from June to September, 2014. 44 teachers from one college were conducted the new dietary intervention as experimental group, 37 teachers from another college were the control group, used the usual method. Compared two groups before and after the intervention of dietary behavior and blood lipid. Results The experimental group′s dietary behaviors changed strongly after implement the new dietary intervention, the scores of DTS before the experiment (58.82 ± 18.47) points,3 months after the experiment (48.36 ± 14.25) points and 6 months after the experiment (44.18±14.92) points were statistically significant (F=21.308, P < 0.01). There was no statistically significant difference in control group (F = 1.129, P > 0.05), respectively (60.51 ± 16.91) points, (57.19 ± 16.35) points, (56.92 ± 21.35) points. After 6 months, the experimental group′s subjects of TC was (4.28±3.73) mmol/L, the control group was (6.23±1.04) mmol/L, the difference was statistically significant (t = 3.082, P < 0.05), there were no significant differences in TG, LDL-C and HDL-C of the two groups (P>0.05). Conclusions The new dietary guidance plan can effectively improve and maintain the dietary behavior in hyperlipidemia college teachers, and decrease the blood lipid level.
7.Effect of Different Electron Donors on Reductive Dechlorination of 2,4-Dichlorophenol
Ting CHENG ; You-Zhi DAI ; Chun-Xiang LUO ; Shuang-Shuang LI ; Wen-Qi TANG ;
Microbiology 2008;0(08):-
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.
8.Clinical analysis of 1272 cases of pelvic endometriosis
Xiaoling ZHANG ; Yufen CHENG ; Shuang WANG ; Qingxian ZHU
Clinical Medicine of China 2010;26(10):1026-1029
Objective To assess the surgical effectiveness of pelvic endometriosis (EMS) by Laparoscopic. Methods Retrospective analysis was performed in 1272 pelvic EMS patients underwent laparoscopic surgical treatment. All patients were classified into four groups according to the R-AFS,63 patients of Ⅰ stage,44 patients of Ⅱ stage,475 patients of Ⅲ stage and 690 patients of Ⅳ stage. Symptoms of EMS,surgical effect and follow-up outcomes were compared. Results In all stages, there were no significant differences on moderate to severe dysmenorrhea,increase in CA125 and infertility (Ps > 0. 05 ). In the EMS patients combined with AM or/and DIE, the rates of the moderate to severe dysmenorrhea, CA125 increase and infertility were 84.6% (11/13 ) ,92.3 %(12/13) and 53.8% (7/13)in stage Ⅰ ,81.8% (9/11) ,90. 9% ( 10/11 )and 54.5% (6/11) in stage Ⅱ ,84.8%(173/204) ,93. 1% (190/204) and 47.1% (96/204) in stage Ⅲ ,and 81.6% (213/261) ,91.2% (238/261) and 46.4% ( 121/261 ) in stage Ⅳ, respectively. The rates in non-consolidated AM or/and DIE patients were 46.0%(23/50),62.0% (31/50) and 22.0% ( 1 1/50) in stage Ⅰ ,45.5 % ( 15/33 ),54.5% ( 18/33 ) and 18.2% (6/33)in stage Ⅱ ,41.7% ( 113/271 ) ,62.7% ( 165/271 ) and 23.6% (64/271) in stage Ⅲ ,and 47.3% (203/429),60.1% (248/429) and 21.7% (93/429). The difference between the two groups with and without consolidated AM or/and DIE had statistical significance( P < 0.05 or P < 0.01 ). After the treatment, the scores of life's energy, ache,emotion,sleep,social activity and body movement of NHP were significantly superior than those before treatment in all patients ( P < 0.01 ). The surgery time-consuming of Ⅲ and Ⅳ stage patients were (64.5 ± 18.4) min and (61.7 ± 17.1 ) min respectively, which were significantly higher than that of Ⅰ and Ⅱ stage ( ( 31.9 ± 12.3 ) min and (40.3 ± 10. 6) main ] ( t = 20.25, P < 0.01 ). The massive hemorrhages and the internal damage organs occurred in Ⅲ and Ⅳ stage surgery [2.9% and 3.5% ,respectively] ,whereas much less in Ⅰ and Ⅱ stage (0.8% and 1.4% ) ,with no significant difference(P >0. 05). After treatment,the rate of symptoms recurrence of Ⅲ and Ⅳ stage patients respectively were 21.4% ,which were higher than that of Ⅰ and Ⅱ stage patients ( 2.3% ) (P < 0.05 )The rate of pregnancy in Ⅲ and Ⅳ stage patients ( 15.4% ) were lower than that of Ⅰ and Ⅱ stage patients (69. 6% ) ( P < 0. 01 ). Conclusions Pelvic EMS decreased the quality of life. EMS patients combined with adenomyosis or/and deep infiltrating EMS have more severe pelvic pain symptoms, less surgical effectiveness and more serious complications. Ⅲ and Ⅳ stage patients are more liable to symptoms recurrence and lower pregnancy rate.
9.Regulation of Moxibustion on the Expression of NF-κBp65 and PPARγ mRNA in Colon of Rats with Ulcerative Colitis
Xiaoming FENG ; Tongbin CHENG ; Huangan WU ; Huirong LIU ; Shuang ZHOU
Journal of Acupuncture and Tuina Science 2014;(5):265-272
Objective: To explore the mechanism of herb-partitioned moxibustion for ulcerative colitis (UC) through observing the colonic mucosal histopathological changes and the expression of nuclear factor kappaB (NF-kB) and peroxisome proliferators-activated receptorγ (PPARγ) mRNA of UC rats. <br> Methods: Male SD rats were randomly divided into a normal group and a model group. UC model was established by general immunological plus local irritation method. After model identification, rats in the model group were randomly divided into a model group, a herb-partitioned moxibustion (HPM) group and a Western medicine (Salicylazosulfapyridine, SASP) group. Rats in the HPM group received treatment at bilateral Tianshu (ST 25) and Dachangshu (BL 25), two cones for each point, once a day for 7 d. SASP group rats were gavaged with SASP. The pathological scores were evaluated according to hematoxylin-eosin (HE) staining of colonic tissues. We used light microscopy to observe degree of colonic mucosal damage and the quantitative polymerase chain reaction (QPCR) to detect the expression of NF-κBp65 and PPARγ in colorectal mucosa. <br> Results: Compared with the normal group, histopathological scores were significantly higher in the model group (P<0.05); compared with the model group, the scores were decreased significantly in the HPM group and the SASP group. Compared with the normal group, NF-κBp65 mRNA expression was increased with statistical significance in the model group (P<0.05); compared with the model group, NF-κBp65 mRNA expressions were decreased significantly in the HPM group and the SASP group. Compared with the normal group, PPARγ mRNA expression was increased significantly in the model group (P<0.05); compared with the model group, PPARγ mRNA expressions were decreased significantly in the HPM group and the SASP group. <br> Conclusion: HPM could improve the mucosa damage of UC rats, which is possibly through down-regulating NF-κBp65 to achieve anti-inflammatory effect. Whether decreasing the PPARγ mRNA is possibly involved in preventing precancerosis will need further study.
10.Significance of Expression of Serum Soluble Vascular Cell Adhesion Molecular and Soluble Intercellular Molecule-1 in Children with Henoch-Schoenlein Purpura
yong, GU ; guo-cheng, XU ; shuang-gen, MAO
Journal of Applied Clinical Pediatrics 2006;0(21):-
Objective To detect the levels of plasma soluble vascular cell adhesion molecular(sVCAM-1) and soluble intercellular mo-lecule-1(sICAM-1) in children with Henoch-Schoenlein purpura(HSP) and its clinical significance.Methods The plasma levels of sVCAM-1 and sICAM-1 were measured by enzyme-linked immunosorbent assay(ELISA) in 53 children with HSP(37 cases in acute stage and 16 cases in recovery stage)and in 25 healthy subjects respectively,and the change that in acute stage and recovery stage was analyzed,at the same time,the 2 factors in children with renal injury and without injury were analyzed.Results The serum levels of sVCAM-1 and sICAM-1 in acute stage HSP children were significantly higher than those in recovery stage and normal controls(Pa