Objective: To clone the full-length cDNA of jasmonate-zim-domain protein (JAZ) gene in Aquilaria sinensis to provide the basic information for further study on gene function in sesquiterpenes biosynthesis pathway. Methods: With the total RNA as template, the full-length cDNA of JAZ in A. sinensis was cloned through rapid amplification of cDNA ends (RACE) technique and reverse transcription PCR (qRT-PCR) method. The bioinformatics of the JAZ gene was analyzed as well. The expression of this gene was detected by qRT-PCR method with MeJA and mechanical wounding treatment in A. sinensis callus. Results: The full-length cDNA (1 507 bp) of JAZ gene was named AsJAZ1; GenBank registration number was KP677281. AsJAZ1 was obtained with an open reading frame (ORF) of 990 bp and encoding 330 amino acids. The relative molecular mass of AsJAZ1 calculated was 34 280, and the isoelecric point was 6.89. Real time PCR results indicated that both MeJA treatment and mechanical wounding could stimulate the increase of mRNA expression of AsJAZ1; There was a sharp rise at 0.5 h with about 27 times higher than the control (without MeJA treatment) with MeJA treatment, then dropped significantly. In mechanical wounding treatment, the highest peak presented in 2 h about 17 times compared to the control, then dropped significantly too. The expression of AsJAZ1 gene returned to be normal in 24 h. Conclusion: We have obtained the full-length cDNA sequence of AsJAZ1 gene firstly, which was extremely sensitive to wounding and responded to the early damage.

Objective: To clone the full-length cDNA of interacting protein of JAZ (NINJA) gene in Aquilaria sinensis, and to provide the basic information for further study on gene function in sesquiterpenes biosynthesis pathway. Methods: With the total RNA as template, the full-length cDNA of NINJA in A. sinensis was cloned through RACE technique and RT-PCR method. The bioinformatics of cloing NINJA gene was analyzed as well. The expression mode of this gene was with MeJA treatment in A. sinensis callus detected by qRT-PCR method. Results: The full-length cDNA (1 982 bp) of NINJA gene in A. sinensis, named as AsNINJA1 was obtained with an open reading frame of 1 221 bp and encoding 406 amino acids. The relative molecular mass of AsNINJA1 protein calculated was 43 697, and the isoelectric point was 6.02. The qRT-PCR results indicated that MeJA treatment could stimulate the increase of mRNA expression of AsNINJA1; There was a sharp rise at 4 h with nearly 100 times higher than the control (without MeJA treatment), then dropped significantly. Conclusion: The full-length cDNA sequence of AsNINJA1 gene is obtained; AsNINJA1 is extremely sensitive to MeJA treatment, and responded to the early damage.

Aim To observe the improvement of visual-auditory cognitive functions in the human entering high altitude by taking in tea polyphenols.Methods Thirty eight males living at 3 700 m high altitudes for 90 days constantly were randomly divided into two groups: ①group Ⅰ(placebo,40 mg/day); ②group Ⅱ(TP,300 mg/day).Cognitive functions were measured by integrated visual and auditory continuous performance test and the difference between groups was evaluated by the comparisons of post-treatment to pre-treatment.Results Compared with pre-treatment,PruA was significantly higher after taking in TP(P

The study aimed to clone the open reading frame of cinnamate 4-hydroxylase (C4H) from Aquilaria sinensis and analyze the bioinformatics and expression of the gene. One unique sequence containing C4H domain was discovered in our previous reported wound transcriptome dataset of A. sinensis. The open reading frame of C4H was cloned by RT-PCR strategy with the template of mixed RNA extracted from A. sinensis stem which treated by different wound time. The bioinformatic analysis of this gene and its corresponding protein was performed. C4H expression profiles in responds to MeJA (methyl jasmonate) application were analyzed by real-time PCR. The length of C4H open reading frame (ORF) was 1 515 bp, encoding 514 amino acids. The GenBank accession number is KF134783. Inducible-experiments showed that the genes were induced by mechanical wound as well as MeJA induction, and reached the highest expression level at 8 h and 20 h, respectively. The full-length cDNA of C4H and its expression patterns will provide a foundation for further research on its function in the molecular mechanisms of aromatic compounds and flavonoids biosynthesis.
Amino Acid Sequence ; Cloning, Molecular ; Models, Molecular ; Molecular Sequence Data ; Open Reading Frames ; Oxidoreductases ; chemistry ; genetics ; metabolism ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Thymelaeaceae ; chemistry ; enzymology ; genetics ; Trans-Cinnamate 4-Monooxygenase ; chemistry ; genetics ; metabolism

OBJECTIVE: To study placenta-derived mesenchymal stem cells with HLA-G (Human Leukocyte Antigen, HLA-G) positive expression induce Treg (regulatory T cell, Treg) in vitro. METHODS: placenta-derived mesenchymal stem cells were separated from neonatal placenta; PEGFP - N1 -HLA-G plasmid was transfected in placenta-derived mesenchymal stem cells by liposome transfection.The cells were divided into 3 groups including control group, PEGFP-N1 group and PEGFP-N1-HLA-G group, 5 complex walls in each group. Expression of HLA-G protein was detected by Western Blotting; after identification of cells, healthy human peripheral blood CD4 T lymphocytes were cultured with placenta-derived mesenchymal stem cells with HLA-G positive expression, and the ratio of CD4CD25Foxp3Treg in T lymphocytes was accounted. RESULTS: After transfection of PEGFP-N1-HLA-G, the placenta-derived mesenchymal stem cells can express HLA-G protein significantly, compared with the control group and PEGFP - N1 group (<0.01). After HLA-G positive placenta-derived mesenchymal stem cells and CD4 + T lymphocytes were cultured for 24 h, the ratio of CD4CD25Foxp3Treg in T lymphocytes was (16.41±0.94)%. After HLA - G positive placenta-derived mesenchymal stem cells and CD4 T lymphocytes were cultured for 48 h, the ratio of CD4CD25Foxp3Treg in T lymphocytes was (16.46±0.59)% significantly, compared with the control group and PEGFP - N1 group (<0.01). CONCLUSIONS Placenta-derived mesenchymal stem cells modified by HLA-G gene can effectively induce CD4CD25Foxp3Treg in vitro.
Female ; Forkhead Transcription Factors ; HLA-G Antigens ; Humans ; Mesenchymal Stem Cells ; Placenta ; Pregnancy ; T-Lymphocytes, Regulatory

OBJECTIVETo investigate the effect of nano-hydroxyapatite/collagen (nHA/collagen) composite as a graft extender and enhancer when combined with recombinant human bone morphogenetic protein-2 (rhBMP-2) on lumbar intertransverse fusion in rabbits.

METHODSSixty-four adult female New Zealand white rabbits, aged 1 year and weighing 3.5-4.5 kg, underwent similar posterolateral intertransverse process arthrodesis and were randomly divided into 4 groups based on different grafts: autogenous cancellous bone alone (ACB group), nHA/collagen alone (HAC group), half autogenous cancellous bone and half nHA/collagen (ACB+HAC group) and nHA/collagen combined with rhBMP-2 (HAC+BMP group). The fusion masses were analyzed by manual palpation, radiography, biomechanical testing and histological examination.

RESULTSFusion was observed in 4 cases in the 6th week and in 5 cases in the 10th week after surgery in ACB group. No case showed fusion in HAC group. In ACB+HAC group, there was fusion in 3 cases in the 6th week and in 4 cases in the 10th week after surgery. In HAC+BMP group, fusion in 1 case was found in the 4th week, in 5 cases in the 6th week and in 6 cases in the 10th week after surgery. It suggested that ACB, ACB+HAC and HAC+BMP groups showed similar fusion ratio and mechanical strength in the 6th and 10th week after surgery. According to the microstructure analysis of the samples, nHA/collagen had no negative effect when implanted together with ilium autograft. In HAC+BMP group, new bone-like tissue was observed in the 2nd week postoperatively, and nearly all of the implanted composites were replaced by mature bone matrix and new bones in 10th week postoperatively.

CONCLUSIONSThe nHA/collagen, especially combined with rhBMP-2, is a promising bone substitute, for it has quick biodegradation, fine bone-bending ability, and high osteoconductivity on posterolateral spinal fusion in rabbits.

Analysis of Variance ; Animals ; Biomechanical Phenomena ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; pharmacology ; Disease Models, Animal ; Durapatite ; pharmacology ; Female ; Lumbar Vertebrae ; drug effects ; surgery ; Osseointegration ; drug effects ; Probability ; Rabbits ; Sensitivity and Specificity ; Spinal Fusion ; methods ; Tensile Strength ; Transforming Growth Factor beta

OBJECTIVETo investigate the feasibility and safety of adult-to-adult living-related donor liver transplantation using a right lobe graft.

METHODSThe clinical data of 2 cases of living-related donor liver transplantation performed between July, 2010 and November, 2010 were analyzed.

RESULTSLiver transplantation was performed using a right lobe graft including the middle hepatic vein in one case and a right lobe graft without the middle hepatic vein in the other. The ratio of graft volume to standard liver volume was 46.2% and 47.3% in the two cases, with GR/WR of 0.83 and 0.80, and donor residue liver of 42.1% and 39.5%, respectively. The donor operation lasted for 6.5 h and 5 h in the two cases with blood loss of about 200-250 ml without blood transfusion. The donors recovered uneventfully without any surgical complications, whose liver function was normal 7 days after the operation, and were discharged 14 days and 16 days after the surgery, respectively. The recipient operation lasted for 8 h and 7 h with blood loss of about 800-1000 ml. The right hepatic vein, hepatic artery, portal vein and bile duct reconstruction were performed by end-to-end anastomoses in the 2 recipients. Bile duct anastomosis stricture occurred in the first recipient 2 months after transplantation and was treated with percutaneous transhepatic cholangiography and drainage. The second recipient recovered smoothly without any complications. The recipients have so far survived 9 months and 5 months, respectively.

CONCLUSIONAdult-to-adult living-related donor liver transplantation is a safe and effective option for treatment of end-stage liver diseases in the context of cadaveric liver graft shortage.

Adult ; Female ; Hepatectomy ; Humans ; Liver Cirrhosis ; surgery ; Liver Neoplasms ; surgery ; Liver Transplantation ; methods ; Living Donors ; Male ; Middle Aged ; Retrospective Studies

The full-length coding sequence (cds) of jasmonate-zim-domain protein (AsJAZ1) gene was cloned from Aquilaria sinensis, the prokaryotic vector was constructed and the recombinant proteins expression was induced to provide the basic material for interactive proteins screen and gene function research. In this study, with the total RNA isolated from A. sinensis leave as template, the full-length cds of AsJAZ1 gene was amplified using RT-PCR method and subcloned into pET-28a vector. The recombinant plasmid identified by restriction enzyme digestion and nucleotide sequencing was transformed into E. coli BL21(DE3). Inducing with 0.5 mmol•L⁻¹ IPTG at 37 ℃ for 4 hours, a fusion protein about 39 kDa was maximumly obtained. AsJAZ1 fusion protein had been expressed successfully mainly in the form of inclusion bodies and only a very small amount was secreted into the cytoplasm in the supernatant.

The MYC2 transcription factor is a member of the important plant bHLH transcription factor families, and it is also the core regulatory elements in jasmonate (JA) signaling pathway. However, there is a little information about AsMYC2 gene in Aquilaria sinensis. In this study, with the total RNA isolated from A. sinensis leave as template, the full-length coding sequence (CDS) of AsMYC2 gene was amplified using RT-PCR method and subcloned into pGEX-4T-1 vector by the gene recombination technique. The recombinant vector pGEX-4T-1-AsMYC2 was verified by restriction enzyme digestion and nucleotide sequencing, and was transformed into E. coli BL21(DE3) to express the protein. A maximum expression of soluble protein was observed with induction by 0.1 mmol·L^-1 IPTG at 37℃ for 4 hours. The fusion protein was purified through a Sepharose-Glutathione column, and verified by SDS-PAGE and Western blotting using an anti-GST polyclonal antibody. We successfully constructed the GST-AsMYC2 plasmid, produced and purified the GST-AsMYC2 fusion protein, which would provide the basic material for polyclonal antibody preparation, interactive factors screening and gene function research. According to the tissue-specific expression pattern analysis by qRT-PCR method, the AsMYC2 gene in A. sinensis tissues is mainly expressed in roots and stems, the main agarwood formation parts, and lowest expressed in leaves. These results indicate that AsMYC2 gene likely play some roles in agarwood formation in A. sinensis.

OBJECTIVEIn mid-July 2005, five patients presented with septic shock to a hospital in Ziyang city in Sichuan, China, to identify the etiology of the unknown reason disease, an epidemiological, clinical, and laboratory study were conducted.

METHODSAn enhanced surveillance program were established in Sichuan, the following activities were introduced: active case finding in Sichuan of (a) laboratory diagnosed Streptococcus suis infection and (b) clinically diagnosed probable cases with exposure history; supplemented by (c) monitoring reports on meningococcal meningitis. Streptococcus suis serotype 2 infection was confirmed by culture and biochemical reactions, followed by sequencing for specific genes for serotype and virulence factors.

RESULTSFrom June 10 to August 21, 2005, 68 laboratory confirmed cases of human Streptococcus suis infections were reported. All were villagers who gave a history of direct exposure to deceased or sick pigs in their backyards where slaughtering was performed. Twenty six (38%) presented with toxic shock syndrome of which 15 (58%) died. Other presentations were septicaemia or meningitis. All isolates were tested positive for genes for tuf, species-specific 16S rRNA, cps2J, mrp, ef and sly. There were 136 clinically diagnosed probable cases with similar exposure history but incomplete laboratory investigations.

CONCLUSIONAn outbreak of human Streptococcus suis serotype 2 infections occurred in villagers after direct exposure to deceased or sick pigs in Sichuan. Prohibition of slaughtering in backyards brought the outbreak to a halt. A virulent strain of the bacteria is speculated to be in circulation, and is responsible for the unusual presentation of toxic shock syndrome with high case fatality.

Animals ; Bacteremia ; epidemiology ; microbiology ; China ; epidemiology ; Disease Outbreaks ; Humans ; Meningitis, Bacterial ; epidemiology ; microbiology ; Shock, Septic ; epidemiology ; microbiology ; Streptococcal Infections ; epidemiology ; microbiology ; veterinary ; Streptococcus suis ; isolation & purification ; Swine ; Swine Diseases ; microbiology