5.Cloing and High Level Expression of VP6 Gene From Group A Human Rotavirus in E.coli
Zhi-Liang CAO ; Yu-Ling WEN ; Yuang-Ding CHEN ;
China Biotechnology 2006;0(03):-
The structural protein VP6 of rotavirus form the middle layer of the triple-layered viral capsid, playing a key role in the organization of the virion. The gene of structural protein 6 of rotavirus strain TB-Chen isolated from a clinic sample was amplified using PCR from the reverse transcription product of RV genome RNA, using pET as expression vector, a recombinant plasmid pET-VP6 containing coding sequence of VP6 protein was constructed. The results showed that the VP6 was highly efficiently expressed in E. coli BL21(DE3) cells which were transformed with the recombinant plasmid pET-VP6.The expressed VP6 protein possessed 27.4% of total cells protein, with an approximately 45kDa of molecular weight, and could be recognized by guinea pig anti-SA11 antibody on Western blot. The results obtained provide important basis for further study on structure and function of the VP6 protein.
6.Effect of Chinese Herbs on Gastrointestinal Motility of Chronic Experimental Beagle Model
Qicheng CHEN ; Fengshun PANG ; Lixing CAO ; Zhi JIANG ; Lyu ZHOU ; Zhiqiang CHEN
Journal of Guangzhou University of Traditional Chinese Medicine 2016;33(5):674-678
Objective To observe the effect of 7 kinds of Chinese herbs(Pericarpium Arecae, Folium Sennae, Fructus Tsaoko, Fructus Amomi, Rhizoma Atractylodis Macrocephalae, Fructus Aurantii, Fructus Aurantii I mmaturus), and 2 kinds of active compounds (bornyl acetate, synephrine) of Chinese herbals on gastrointestinal motility of chronic experimental beagle model. Methods Six beagles were used for inducing chronic experimental model. The beagles’antrum, duodenum, jejunum, ileum and colon were implanted with strain gauges to record canine gastrointestinal motility. Gastric fistula was set up for the intake of Chinese medicine decoction, and the external jugular vein catheter was made for intravenous administration. After modeling, the fasting gastrointestinal motility features of the beagles were observed for 120 min every day, and for 7 continuous days. From the 7th day after modeling, fasting gastrointestinal motility before medication was recorded as fundamental control, and when the interdigestive migrating motor complex (MMC) I phase arrived, the gastrointestinal motilities were sequently recorded after treatment with the 7 kinds of herbs(gavage of 200 mL of the decoction of each herb through gastric fistula), normal saline(200 mL), bornyl acetate(active ingredient of Fructus Aurantii and Fructus Aurantii Immaturus, intravenous injection), and synephrine (active ingredient of Fructus Amomi, intravenous injection). All of the animals were treated with only one kind of Chinese herb one day , and the observation of each herb lasted for 2 continuous days. MMC cycle, frequency of contraction, sum of contraction, amplitude of contraction, average of amplitude, and motor index (MI) were observed by strain gauges. Results On postoperative day 1-6, the animals had gastrointestinal hypomotility and no MMC cycle was recorded. On postoperative day 7, the canine antrum, duodenum, jejunum and ileum showed typical MMC cycle while the colon had irregular MMC cycle at fasting interdigestive period. Compared with the gastrointestinal motilities before treatment and those of the normal saline group, the number of contraction waves and MI were significantly increased in the antrum, duodenum, jejunum, ileum and colon after intervention with Pericarpium Arecae and bornyl acetate, were increased in the duodenum, jejunum, ileum and colon after treatment with Folium Sennae and Fructus Tsaoko (P<0.05), and were also enhanced in the antrum and duodenum after administration of Fructus Amomi(P < 0.05). Treatment with Fructus Aurantii induced the decrease of MI in the jejunum, Fructus Aurantii Immaturus decreased MI of the jejunum and colon, and synephrine decreased the number of antrum contraction waves and MI of the antrum and jejunum(P < 0.05). Conclusion The chronic experimental model is effective for the screening of Chinese herbs for improving gastrointestinal motility. Pericarpium Arecae, Folium Sennae, Fructus Tsaoko, Fructus Amomi, and bornyl acetate can increase gastrointestinal motility, while Fructus Aurantii, Fructus Aurantii Immaturus and synephrine can inhibit the gastrointestinal movement, and Rhizoma Atractylodis Macrocephalae has no effect on gastrointestinal motility.
7.Effects of genistein on PCNA expression and cell cycle in human hypertrophic scar fibroblasts in vitro
Chuan CAO ; Shirong LI ; Heng YAO ; Zhi FENG ; Xia DAI ; Yanqing CHEN ; Xiaoge LI ; Liang CHEN
Chinese Journal of Medical Aesthetics and Cosmetology 2008;14(3):159-162
Objective To observe the effects of genistein on PCNA expression and cell cycle in fibroblasts derived from human hypertrophic scar in order to explore the mechanism of its inhibition on hypertrophic scar (HS) fibroblast proliferation. Methods The human hypertrophic scar fibroblasts were cultured in vitro. Genistein with various concentrations (25, 50, 100 μmol/L) was co-cultured in the medium for 48 hours. The expression of PCNA was detected with immunocytochemical staining method and the cell cycle was measured with flow cytometry. Results Genistein could significantly decrease PCNA expression in HS fibroblasts, especially when its concentration at 50 μmol/L or 100 μmol/L. The cell percentage of G0~G1 phase decreased with drug′s concentration, and G2~M percentage increased conversely, implying the suspension of mitosis. In 100 μmol/L group, most cells blocked at S phase and a hypodiploid apoptosis peak could be observed ahead of G1 phase. Conclusion Genistein can inhibit the proliferation of human hypertrophic scar by blocking cell division as well as decreasing DNA synthesis.
9.Screening of Poly-?-glutamic Acid High Productive Strain and Optimization of Fermentation Medium
Xue-Chao WU ; Xin-Jiang CAO ; Zhi-Xia JI ; Shou-Wen CHEN ;
Microbiology 2008;0(10):-
Bacillus subtilis B6-1 was used as an original strain for mutagenic treatment and a defined medium was used as the selective medium. A mutant named B. subtilis W003 was isolated after three serial ultraviolet (UV) irradiations and one diethyl sulfate (DES) treatment. The ?-PGA yield on a rotary shaker was enhanced from 10.9 g/L in parental strain to 20.5 g/L in the mutant. It was illustrated by single factor experiments that the optimal carbon and nitrogen sources were glucose and (NH4)2SO4 respectively. The optimal fermentation medium was achieved by orthogonal test. In the optimal medium, a ?-PGA yield of 45.3 g/L was obtained after 36 h cultivation.
10.Significance of Detection of Allergen-Specific Serum Immunoglobulin E in Children with Asthma
xin, SONG ; wei, WANG ; jing, ZHAO ; ling, CAO ; yu-zhi, CHEN
Journal of Applied Clinical Pediatrics 2004;0(09):-
Objective To investigate the positive rates of food allergen (fx5),mixed molds (mx1),phadiatop and house dust mites (d1) allergen-specific serum immunoglobulin E(sIgE) in children with asthma or cough variant asthma (CVA).Their age distribution characte-ristics were also compared and analysed.Methods Blood samples of 597 asthmatic children aging from 6 months to 16 years old were applied to fx5,mx1,pha-diatop and d1 test by using fluoroenzyme-immunometricassay,UniCAP100.The positive rate of fx5,mx1,phadiatop and d1 were compared.Age distribution characteristics of children with positive sIgE were analysed.Results 1.The positive rate of fx5,mx1,pha-diatop and d1 were 34.6%,30.0%,35.2% and 23.5%,respectively.2.The highest positive rate of phadiatop and d1 were highest in children of 6-16 age group(67.8% and 46.3%);The positive rate of fx5,mx1 peaked in children of 2 years and 5 years separately(44.1% and 53.6%);3.Among 181 children with mx1 positivity 90 cases manifested phadiatop negativity.4.Sixty-five percent of children with clinically diagnosed asthma or CVA presented positivity in test in fx5,mx1 or phadiatop at least.Conclusions Two thirds asthma and CVA patients exist sIgE.Asthmatic infants mainly manifest food allergen sensitization.The positive of phadiatop increased with age.Positivity of mx1 increased with age ≤5 years old in children,and it decreases in older children(≥6 years old).Mx1 is one of the main allergens in asthmatic children.