1.Effect of rare-earth elements La3+ on growth of hairy roots and normal roots of Rheum palmatum and their yield of anthraquinone
Shihai YANG ; Xiaofeng LIU ; Dean GUO
Chinese Traditional and Herbal Drugs 1994;0(10):-
Object To study the effect of rare-earth elements La3+ on growth of hairy roots and normal roots of Rheum palmatum L and their yield of anthraquinones. Methods The biomass and an- thraquinone yield of two hairy clones DH5c, DH7a, and normal root NOR cultures of R. palmatum were evaluated statistically after various concentration of La3+ were administrated into culture medium. Results Significant differences of biomass yield and anthraquinone yield were shown among six La3+ concentrations, in which 10 mg/L showed obvious inhibitory effect for root growth and 1. 0 mg/L was the best for anthraquinone yield. In general, aloe-emodin and rhein were predominant in five anthraquinones after treated with the rare-earth element. Conclusion The growth of hairy root cultures of R. palmatum and their anthraquinone yield are greatly promoted by addition of La3+ to the medium.
2.Biotransformation of podophyllotoxin by cell suspension and root culture systems of Rheum palmatum
Shihai YANG ; Jiayuan LIU ; Bo YU ; Xiaofeng LIU
Chinese Traditional and Herbal Drugs 1994;0(05):-
Objective To investigate the biotransformation of podophyllotoxin by cell suspension culture and root culture systems of Rheum palmatum.Methods Plant tissue culture technology was employed.The products were isolated by silica gel column chromatography and their structures were elucidated by spectral methods.Results Adding of podophyllotoxin had no obvious effects on the pH value of cell suspension culture and root culture systems of R.palmatum.In the case of present study,73.8% of podophyllotoxin was converted by cell suspension culture and 56.3% by root culture.The products were different between the two culture system.Podophyllotoxin was converted into picropodophyllin with cell suspension culture system and many other products besides epipodophyllotoxin with root culture system.Conclusion Both of cell suspension culture and root culture systems of R.palmatum are able to convert podophyllotoxin.
3.Ischemic precondition inhibits pancreatic acinar cells apoptosis in rats with ischemia/reperfusion injury following pancreas transplantation: Role of reactive oxygen and mitochondrial DNA repair enzyme
Yiling HOU ; Hai BO ; Ziquan LIU ; Shihai XIA
Chinese Journal of Tissue Engineering Research 2010;14(18):3279-3285
BACKGROUND: Ischemic preconditioning (IPC) can induce endogenous protection mechanism, which effectively prevent ischemia/reperfusion injury following organ transplantation. Cold and warm ischemia may induce ischemia/reperfusion injury of pancreas transplantation, and apoptosis of pancreatic acinar cells is one of the important reasons of pancreas graft functional defect after transplantation. Mitochondrial DNA has repair system, and its balance with mitochondrial DNA injury influences disease occurrence and outcome.OBJECTIVE: To observe the effect of IPC on apoptosis of transplanted pancreatic acinar cells, and the possible role of reactive oxygen (ROS) and mitochondrial DNA repair enzyme.METHODS: A total of 50 health, male, Sprague-Dawley rats were randomly divided into three groups: sham operated (n = 10), donors (n = 20) and recipients (n = 20). The recipients were randomly divided into ischemia/reperfusion group (IR, n = 10) and IPC group (n = 10). The sham operated group was subjected to abdominal open and close operation. IR group and IPC group received establishment of diabetic model by streptozotocin injection. IR rats received whole pancreatic-duodenal transplantation alone. IPC rats received whole pancreatic-duodenal transplantation exposed ischemic preconditioning with 5 minutes ischemia and 5 minutes reperfusion twice. All grafts were keep with warm ischemia time 15 minutes and cold ischemia (in 4 ℃ UW preservation solution) time 180 minutes. Twelve hours after reperfusion, serum amylase, blood glucose, Caspase-3, -9 activity were detected. Pancreatic acinar cell apoptosis was measured by flow cytometry. Mitochondrial cross-membrane potential (Δψ) was measured by monitoring the fluorescence spectrum of rhodamine 123. Mitochondrial H2O2 generation was determined using dichlorofluorescein as a probe. 8-oxodG in mitochondrial DNA (mtDNA) was measured with HPLC system. Release of cytochrome C, phosphorylation of Akt and mitochondrial OGG1 protein expression were determined by Western-blotting. RESULTS AND CONCLUSION: The ischemia preconditioning can relieve the pancreatic acinar cell apoptosis in pancreas graft and relieve IR injury by decreasing mitochondrial oxidative stress, mtDNA injury, and increasing phosphorylation of Akt and mitochondrial OGG1 expression.
4.Study on Bi-Min-Gan Granules for Regulation of Allergic Rhinitis Treg Cells Induced Immune Tolerance
Jun SHI ; Yu LIU ; Shihai YAN ; Daonan YAN
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;17(4):841-845
This study was aimed to exploreBi-Min-Gan (BMG) granules induced immune tolerance for allergic rhinitis cases through changes of Treg cells to achieve the goal of treatment. SD rats were used as experimental animal, which were divided into the blank group, model group and BMG granules group. Allergic rhinitis rat model was established. Intragastric administration of BMG granules was given. Levels of CD4+CD25+Foxp3+Treg and IL-4, IL-10, TGF-β1, IL-13 were detected with flow cytometry and ELISA assay. The results of flow cytometry showed that CD4+CD25+Foxp3+Treg decreased after the successful establishment of the model (P<0.05); but it increased after treatment of BMG granules (P<0.05) to reach the level of the normal blank group (P>0.05). ELISA showed the concentrations of IL-10 and TGF-β1 decreased significantly after the successful establishment of the model (P<0.0001); concentrations of IL-4 and IL-13 increased significantly (P<0.0001). While, after treatment of BMG granules, concentrations of IL-10 and TGF-β1 in blood rebound but did not reach the level of the normal blank group;concentrations of IL-4 and IL-13 in the blood decreased but did not reach the level of the normal blank group. It was concluded that BMG granules can upregulate the number of Treg cells in allergic rhinitis rat, increase the secretion of IL-10 and TGF-β1, inhibit Th2 cell activity, and reduce secretion of IL-4 and IL-13, so as to improve symptoms of allergic rhinitis.
5.Integron-mediated multidrug resistance in clinical multi-resistant Pseudomonas aeruginosa
Peiming LIU ; Huilin YAO ; Shihai LU ; Nan ZHUANG
Chinese Journal of Clinical Laboratory Science 2006;0(03):-
Objective To investigate the action of integron-mediated multidrug resistance in clinical multi-resistant Pseudomonas aeruginosa in Huaibei area.Methods The antibiotic susceptibility of 13 different antibiotics of 36 multi-drug resistant P.aeruginosa was tested by K-B.The three-dimensional method was taken to differentiate the various beta-lactamases.The integron was determined by PCR with integron-specific primer.Results The positive rate of integron detection in multi-resistant Pseudomonas aeruginosa was 86.1%.Two kinds of integron with different length were found,i.e.1 000 bp and 2 000 bp.PSE-1,aadA2,aadB,aac(6)-II and CARB-8 were the resistance-related gene cassette in the integrons.Conclusions Integron may enroll in multiple resistance of multi-resistant Pseudomonas aeruginosa.
6.?-Lactam Genes in Clinical Isolates of Pseudomonas aeruginosa
Peiming LIU ; Huilin YAO ; Shihai LU ; Shuguo QIN ; Yuanhong XU
Chinese Journal of Nosocomiology 2004;0(10):-
OBJECTIVE To investigate the genes associated with the drug-resistance to ?-lactam antibiotics in Pseudomonas aeruginosa(PAE) isolated from clinical patients in Huaibei,Anhui Province.METHODS Antibiotic sensitivity test was performed by VITEK-AMS.The three-dimensional method was taken to differentiate various beta-lactamases.The ?-lactamases and the outer membrane protein D2(oprD2) genes were detected by polymerase chain reaction(PCR) in 36 PAE strains.RESULTS In 36 PAE strains,the positive rates of genes of TEM,DHA,CTX-Ge and CARB were 77.8%,69.2%,27.8% and 25.0%,respectively,that of the OXA-1,OXA-10,VIM-2 and IMP-1 were 16.7%,16.7%,13.9% and 2.8%,respectively.The rate of lacking oprD2 gene was 58.3%,and none possessed VEB,SHV,GES,PER,GIM and SPM-2 genes.CONCLUSIONS P.aeruginosa carries various beta-lactamase genes in clinical patients of Huaibei,and the deletion ratio of oprD2 gene is higher.
7.Three ceramides from gorgonian Echinogorgia sp.
Liu LIAO ; Nan WANG ; Qiu LIANG ; Xiaojian LIAO ; Shihai XU
Chinese Traditional and Herbal Drugs 1994;0(06):-
Objective To find new compound from gorgonian.Methods Three ceramides have been isolated from the South China Sea gorgonian Echinogorgia sp.by silica gel column chromatography.Results Their structures were established as(2S,3S,4R)-N-[2-(1,3,4-trihydroxyicosan-2-yl)]-hexadecanamide(1),(2S,3S,4R)-N-[2-(1,3,4-trihydroxyicosan-2-yl)]-heptadecanamide(2),and(2S,3S,4R)-N-[2-(1,3,4-trihydroxyicosan-2-yl]-octadecanamide(3) by spectroscopic methods and chemical conversion.Conclusion It is the first time to report the three chemical compounds from coral Echinogorgia sp.and compound 2 is a new compound.
8.Study on sealing ability of iRoot used as root end retrograde filling material
Li LI ; Jia LIU ; Shihai YIN ; Wanshan LI ; Leixi REN
Chongqing Medicine 2017;46(22):3097-3100
Objective To detect the sealing ability of iRoot as root end retrograde filling material in vitro experiment,and to compare it with mineral trioxide aggregate(MTA) in order to provide reference basis for clinical use of iRoot.Methods The dental crown was removed in 67 single root canal anterior teeth.The routine root canal preparation and filling were performed.The root end was resected by 3 mm for conducting root end retrograde preparation.Sixty-three teeth in the samples were randomly divided into 4 experimental groups.Then the different materials were used for root end filling:iRoot BP (group A,n=17),iRoot BP plus (group B,n=17),MTA (group C,n =17),and group D(iRoot BP + gutta percha,n =12),the residual 4 samples served as the control group.The apical microleakage of 10 teeth in each experimental group was detected by the fluid dynamics.The microleakage situation at the time points of 1,7,14,28 d was observed.In addition,2 teeth selected from each group were performed the filling material ultramicrostructure observation by SEM.Each 5 teeth were remained in the group A,B and C and performed the Micro CT scanning.The porosity rate of filling material was detected.Results No obvious edge gap was found between the filling materials and dentin surface in the group A,B and D by SEM,but obvious gap could be seen in various samples of group C.Micro CT scanning found that the porosity rates in the group A,B and C were 0.020 ±0.023,0.045 ±0.035 and 0.031 ± 0.011 respectively,but showing no statistical difference(P>0.05).The fluid dynamic experiment showed that the microleakage detection value in each group had no statistical difference(P>0.05).Conclusion The sealing ability of iRoot and MTA used in root end retrograde sealing has no obvious difference,But iRoot operability is superior to MTA.
9.Effect of phytohormones and elicitors on growth and anthraquinone production of Cassia obtusifolia hairy roots
Shihai YANG ; Xiaofeng LIU ; Dean GUO ; Junhua ZHENG
Chinese Traditional and Herbal Drugs 1994;0(05):-
Objective To study the effects of phytohormones and elicitors on the growth and anthraquinone production of Cassia obtusifolia hairy roots. Methods Blades and petioles of C. obtusifolia〖WTBZ〗 were infected with Agrobacterium rhizogenes LBA9402 and the in vitro culture system of the hairy root was established. Results Hairy roots were induced by A. rhizogenes LBA9402. The results showed that the addition of 0.5% NAA to the hairy roots increased the content of five kinds of anthraquinones by four times, and Aspergillus niger elicitor had no much effect and JA had slightly inhibitory effect on root growth, but both of them significantly promoted the anthraquinone accumulation, the anthraquinone yield increased more than two times with the treatment of 2.0 mL elicitor/50 mL medium and by 73% with the treatment of 10 ?mol/L jasmin acid (JA), respectively. JA had little inhibitory effect on the growth of fairy roots. Conclusion The fundation has been established for further oplimizing the proper cultural system for C. obtusifolia hairy roots and regulating the secondary melabolite.