The shortage of human hearts has brought the current research focus on finding an animal source as substitute hearts.The immunological barriers to cardiac xenotransplantation are now more clearly defined,allowing retrospective interpretation of past clinical experience in humans.Due to physiological compatibilities as well as ethical and infectious considerations,pigs have now emerged as the most likely source of future xenografts.With the introduction of transgenic pigs expressing human complement regulatory proteins and new immunosuppressive strategies have shown early promise in the laboratory,cardiac xenotransplantation has been the social focus.This article explores ethical issues that surround developments in this field.

OBJECTIVETo clone and sequence the cDNA of squalene epoxidase gene in Eleutherococcus senticosus.

METHODTotal RNA of E. senticosus was extracted by the improved isothiocyanate method and reverse transcripted into cDNA. The primers were designed depending on the reported SE cDNA sequences of Panax ginseng. The SE cDNAs in E. senticosus was amplified using RT-PCR strategy.

RESULTSequencing results showed two different cDNA fragments (SE1, SE2) with 1665, 1629 bp each ORF which encoded 554,542 amino acids, respectively. The identities of nucleotides and amino acids between SE1, SE2 were 91.49%, 92.55%. SE1, SE2 had the highest amino acids similarity to the SE1 of P. notoginseng, 93.45%, 94.87% respectively. SE1, SE2 both had a FAD binding domain. The deduced speculated amino acids of SE1, SE2 each had 2,4 membrane-spanning helices.

CONCLUSIONThe two SE sequences in E. senticosus were firstly separated and reported, which has made foundation for E. senticosus secondary metabolite engineering researches.

Amino Acid Sequence ; Cloning, Molecular ; DNA, Complementary ; chemistry ; genetics ; Eleutherococcus ; enzymology ; genetics ; Isoenzymes ; classification ; genetics ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Squalene Monooxygenase ; classification ; genetics

OBJECTIVETo analyze the effect of endophytic fungi on expression amount of key enzyme genes SS (squalene synthase gene), SE (squalene epoxidase gene) and bAS (beta-amyrin synthase gene) in saponin biosynthesis and saponins content in Eleutherococcus senticosus.

METHODWound method was used for back meeting the endophytic fungi to E. senticosus. With GAPDH as internal control gene, the expression of key enzyme genes was detected by real time PCR method. E. senticosus saponins content was measured by spectrophotometry method.

RESULTWhen wound method back meeting P116-1a and P116-1b after 30 d, the expression content of SS improved significantly (P < 0.05), however the back meeting of P109-4 and P312-1 didnt change the expression of SS. After that SS expression showed reduction-equality-reduction varying trend. Thirty days after back meeting P312-1, the expression content of SE improved significantly (P < 0.05). Ninty days after back meeting P116-1b and P312-1, the expression content of SE improved significantly to 130%,161%, respectively (P < 0.05). After 120 d, back meeting four endophytic fungi, the expression of SE were significantly higher than the control (P < 0.05). Back meeting four endophytic fungi form 60 d to 120 d, the expression of bAS was significantly higher than the control (P < 0.05). The back meeting four endophytic fungi improved E. senticosus saponins content significantly (P < 0.05).

CONCLUSIONEndophytic fungi P116-1a, P116-1b, P1094 and P312-1 significantly effected the expression of key enzyme genes SS, SE and bAS and then affected E. senticosus saponins content. Among the genes, bAS was key target gene.

Eleutherococcus ; chemistry ; metabolism ; microbiology ; Endophytes ; physiology ; Farnesyl-Diphosphate Farnesyltransferase ; genetics ; Fungi ; physiology ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Plant ; Intramolecular Transferases ; genetics ; Saponins ; analysis ; biosynthesis ; Squalene Monooxygenase ; genetics

OBJECTIVETo clone farnesyl diphosphate synthase (FPS) gene from Eleutherococcus senticosus and analyze the bioinformatics and expression of the gene.

METHODThe FPS full length cDNA was cloned by rapid amplification of cDNA ends (RACE). The data was analyzed by bioinformatics method, the structure and function of FPS was deduced. The expression of FPS in different organ of E. senticosus was detected by RT-PCR.

RESULTThe full length of FPS cDNA was 1 499 bp containing a 1 029 bp ORF that encoded 342 amino acids. The deduced protein sequence exhibited two Asp riches conserved motifs (DDXXD). Without transmembrane domain, FPS was located in cytoplasm. RT-PCR result showed that FPS gene expressed in different organs of E. senticosus. The expression amounts of FPS in different organs were different significantly (P < 0.05).

CONCLUSIONThe FPS gene of E. senticosus was successfully cloned for the first time, and provided a stable foundation for studying on its effect and expression control on E. senticosus saponins biosynthesis.

Amino Acid Sequence ; Cloning, Molecular ; Computational Biology ; Conserved Sequence ; Eleutherococcus ; enzymology ; genetics ; Gene Expression Regulation, Plant ; Geranyltranstransferase ; chemistry ; genetics ; metabolism ; Models, Molecular ; Molecular Sequence Data ; Phylogeny ; Protein Conformation

OBJECTIVETo clone calmodulin (CaM) gene in Eleutherococcus senticosus, and study the effect of endophytic fungi on expression amount of CaM gene.

METHODThe CaM full length cDNA sequence was cloned by rapid amplification of cDNA ends (RACE). The gene was analyzed and corresponding structure and functions were predicted by the bioinformatics methods. The expression amount of CaM gene affected of endophytic fungus P116-1a, P116-1b, P1094 and P312-1 was detected by RT-PCR.

RESULTThe full length of CaM cDNA was 856 bp containing an ORF of 450 bp that encoded a protein of 149 amino acids. The homologous of predicted protein was almost 100% with plants like Panax ginseng and Daucus carota. RT-PCR results showed that endophytic fungus improved CaM expression amount significantly (P<0.05). The highest expression amount of CaM occurred 90 d after reinoculated with endophytic fungi P1094, up to 2.96 times of the control.

CONCLUSIONThe CaM gene of E. senticosus was successfully cloned for the first time. The results demonstrated that endophytic fungus of E. senticosus improved CaM expression amount significantly.

Calmodulin ; chemistry ; genetics ; metabolism ; Cloning, Molecular ; Eleutherococcus ; classification ; genetics ; metabolism ; microbiology ; Endophytes ; physiology ; Fungi ; physiology ; Gene Expression Regulation, Plant ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism

Objective:To investigate the feasibility and safety of a new method of natural orifice transluminal endoscopic surgery(NOTES) -totally transtracheal endoscopic thyroidectomy(TTET) .Methods:Three miniature swines and 6 beagle dogs were underwent TTET.Under general anesthesia,special designed endotracheal tube with 2-channel was used and endoscope and instruments were inserted through the respective channel.Incision of tracheal anterior wall was accomplished and partial or subtotal thyroidectomy was performed.Finally,the defects in the trachea were sutured with ENDO STITCH instrument.Results:Partial thyroidectomy was successfully accomplished on 3 pigs and subtotal thyroidectomy was done on 6 dogs.No serious complications such as anoxia,asphyxia,airway obstruction and death occurred during the operation.Animals were sacrificed 2h after the procedure and incision of trachea was found to be closely sutured.There were no subcutaneous emphysema and haematoma formation.Conclusion:Preliminary experimental results showed the feasibility and safety of TTET.Transtracheal access maintains the integrity of cervical tissues and achieves an optimal cosmetic outcome.TTET may open up a new field of NOTES on thyroid surgery.

OBJECTIVETo reveal the influence of harvest season and tree age on the content of taxol and 10-DAB III, and provide the basis for the harvest time of Taxus madia.

METHODBranches and leaves of the labeled 25 three-year-old plants and 25 five-year-old plants were collected every two months from March 26,2009 to January 26, 2010. Taxol and 10-DAB III content of different age and growth season of Taxus in branches were determined by HPLC.

RESULTTaxol and 10-DAB III content were significantly different in different harvesting age. The content of five-year-old plants was significantly higher than that of three-year-old plants. Taxol and 10-DAB III contents were significantly different in different harvesting season, and the highest content of taxol and 10-DAB III was 0.56, 0.32 mg x g(-1), respectively, in May.

CONCLUSIONThe May is the suitable harvest season for T. madia, but the suitable harvest age need further study which according to the main active component and biomass accumulation.

Antineoplastic Agents, Phytogenic ; analysis ; pharmacology ; Asteraceae ; chemistry ; Chromatography, High Pressure Liquid ; Paclitaxel ; analysis ; pharmacology ; Plant Preparations ; pharmacology ; Seasons ; Taxoids ; analysis ; pharmacology ; Taxus ; chemistry ; drug effects ; Trees ; chemistry