The leucoanthocyantin reducase (LAR) gene, an important functional gene of catechins biosynthesis pathway, was cloned from Fagopyrum dibotrys (D.Don) Hara by degenerate PCR and rapid amplification of cDNA ends (RACE). The full-length cDNA of FdLAR is 1 581 bp (GenBank accession: JN793953), containing a 1 176 bp ORF encoding a 391 amino acids protein, and its 3'-untranslated region has an obvious polyadenylation signal. The recombinant plasmid containing FdLAR completed ORF was transformed into E. coli BL21 (DE3). The target fusion peptide with molecular weight of 66 kD was expressed under the condition of 16 degrees C and induced by IPTG at final concentration of 1.0 mmol x L(-1). Bioinformation analysis indicated that the amino acid sequence of FdLAR showed great homology to other LAR with the NADB-Rossmann conversed domain in the N-terminus. Real-time quantitative PCR was used to detect the expression levels of FdLAR gene during different development periods. The determination of flavonoids contents in appropriate rhizomes showed that the relationship between FdLAR gene expression and the accumulation of flavonoids displayed different trends during vegetative growth and reproductive growth stages, suggesting that the FdLAR gene may be involved in the pathway of flavonoid metabolisms in Fagopyrum dibotrys.

To undertake retrospective analysis of the research and advancement of treating avascular necrosis of the femoral head.After comparing the superiority and inferiority of different treatments and the present therapeutic status many therapeutic methods for avascular necrosis of the femoral head have been performed,commonly according to the staging of necrosis.Conservative therapy is suitable for stage 0 ～Ⅰ,interventional therapy is suitable for stage Ⅱ～Ⅲ,operation is adapted for stage Ⅱ～ Ⅲ and femoral head collapse or degenerative changes.Avascular necrosis of the femoral head is a chronic and dysfunctional illness.Comprehensive treatment according to different stage is now the most popular.Interventional therapy is the study focus of the avascular necrosis of the femoral head meanwhile.(J Intervent Radiol,2006,15: 636-639)

Objective:To evaluate the physiologic work of breathing (WOBphy)as parameter of respiratory weaning and extubation. Method: Patient work of breathing (WOBt) and imposed work of breathing (WOBimp) were measured. WOBphy was obtained with WOBt minuting WOBimp. In the patients who did not meet conventional respiratory weaning parameters ,if WOBphy

Objective To explore the effect of hepatocyte growth factor on peripheral nerve regeneration. Methods Sciatic nerve contusion injury was made by a custom-made clamp in Wistar rats,in which human hepatocyte growth factor expressed by adenoviral vector(Ad-HGF)was injected into the muscle around the injured nerve.The results of nerve regeneration were evaluated by sciatic nerve function index(SFI),muscle wet weight,neural electrophysiology and image analysis. Results Four weeks after sciatic nerve injury,the results of sciatic nerve function index(SFI),muscle wet weight,neural electrophysiology and image analysis showed better nerve regeneration in group injected with HGF than control group(P<0.05). Conclusion Hepatocyte growth factor can promote axon regeneration and functional recovery and is an effective neurotrophic factor for peripheral nerve regeneration after injury.

Objective To investigate the antitumor immunity induced by tumor derived mixed heat shock protein/peptide(mHSPs),interleukin-12(IL-12)and Cyclophosphamide(Cy).MethodsPurified mixed HSP was prepared from tumor by S180 protein extraction and purification,SDS-PAGE,Western blot and animal experiment were applied for mixed HSPs analysis.ResultsThe proliferation of cytotoxic T lymphocyte(CTL)cocultured in the mHSPs+Cy+IL-12 group was significantly remarkable and the content of CD8+ CTLs was significantly more in comparison with the other groups(P<0.01).To the tumor bearing mice,mHSPs+Cy+IL-12 group showed partial therapeutic efficacy,the averaged survival period was over 60 d,and 90% of the mice in this group got long period tumor free survival(>90d),obvious difference(P<0.05)from the other groups.ConclusionTumor derived mixed HSPs can induce powerful antitumor immune efficacy and show favorable therapeutic efficacy.

BACKGROUNDIn order to enhance the chemotherapeutic efficacy of advanced lung cancer and to practise the individualized treatment, it is necessary to find out the difference of anticancer drug-sensitivity in lung cancer cells. Comparing the array profile of related gene of anticancer drug-sensitivity between non-small cell lung cancer (NSCLC) cell lines and immortal human bronchial epithelial cell line BET2A, the difference of expression of related genes of anticancer drug sensitivity was detected.

METHODSWith the technique of cDNA macroarray, the different related gene proceeding of anticancer drug sensitivity expression was analysed in 6 NSCLC cell lines and BET2A cell line. RT-PCR was used to reconfirm the results.

RESULTSSeventy-three genes which were differentially expressed were found from 1291 candidate genes, and there were 45 genes upregulated, and 28 genes downregulated. The results of RT-PCR were consistent with those of cDNA macroarray.

CONCLUSIONSThe main reason of different sensitivity might be the difference of related gene of anticancer drug sensitivity expression. The results of this study provide new targets for reversing multiple drug-resistance, and they also provide experimental evidence to develop some new drugs and to realize the individualized treatment clinically.

Objective To probe the immunological traits of mesenchymal stem cells derived from umbilical cord Wharton's jelly (WJMSCs). Methods The diced Wharton's jelly which was from healthy fullterm birth human umbilical cord was cultured. The mesenchymal stem cells were identified with mesenchymal stem cells markers expression by flow cytometry and multiple differentiation ability. The expression of MHC- Ⅰ / Ⅱ, costimulatory molecules (CD40, CD80 and CD86) was detected with flow cytomctry, immunocytochemistry, and RT-PCR. The expression of immune inhibitors like HLA-G, IDO, and PGE2 was detected by immunocytochemistry and RT-PCR. The expression of immune-related molecules as IL-10, TGF-β, FGF and VEGF was detected with antibody microarray and western blot. Further more, to clarify the in vivo immune reaction of hWJMSCs, we fabricated the hWJMSC-scaffold constructs and implanted them into the rabbit backs. The lymphocyte infiltration and implanted cell survival observed with immunofluorescence. Results After culturinge of diced Wharton's jelly tissue, we obtained spindle-shaped cells. With differentiation medium, the cells can differentiate into osteoblasts, chongdrocytes, adipose cells and schwann cells. Expression of MHC, costimulatory molecules, and a series of immune suppressive-related molecules was found. Immune inhibitors as HLA-G, 1DO, PGE2, and immune suppressive related molecules as HGF, VEGF, TGFand IL-10 were positively expressed. But the cells did not express MHC-Ⅱ. No immune rejection was observed in vivo after implantation of hWJMSC-scaffold constructs. Conclusion It can be concluded that hWJMSCs have very low immunogenicity, which means the cells have potential to induce immune tolerance.The hWJMSCs do not provoke immune rejection in vivo.

OBJECTIVETo get aesthetic implant-supported restorations by means of the all-ceramic crown fused to small-size titanium abutment and evaluate the quality of the restorations.

METHODSA small-size titanium abutment that had extremely thin circumferential collar and axial wall was fabricated, while a Cercon all-ceramic crown made with computer aided manufacture (CAM). The crown was fused to the abutment by firing the opaque porcelain. A total of 6 restorations in 5 patients were installed and assessed according to the California Dental Association (CDA) quality evaluation system.

RESULTSAll the restorations were fabricated well and ranked in clinic evaluation excellent for surface, anatomical form, marginal quality, and color at baseline and one year after insertion, respectively.

CONCLUSIONSThe restoration of all-ceramic crown fused to titanium small-size abutment is a new aesthetic alternative for the implant-supported restoration.

Adult ; Crowns ; Dental Abutments ; Dental Implants, Single-Tooth ; Dental Porcelain ; Dental Prosthesis Design ; Dental Prosthesis, Implant-Supported ; methods ; Female ; Humans ; Male ; Titanium

OBJECTIVETo observe the time course of proliferation and differentiation of neural stem cells (NSCs) in the subventricular zone (SVZ) of rats following traumatic craniocerebral injury (TBI).

METHODSForty-eight SD rats were randomized into 3 groups, namely the control group without any treatment, the sham-operated group with scalp incision and preparation of a cranial window, and TBI group with craniocerebral injury induced by Feeney's method. With nestin and BrdU as two cell markers, NSE as the neuron-specific marker and GFAP as the glial cell marker, immunofluorescence assay with double labeled antibodies was performed to examine the proliferation and differentiation of endogenous NSCs in the SVZ at different time points after TBI.

RESULTSs The numbers of cells positive for nestin/NSE, nestin/GFAP, BrdU/NSE, and BrdU/GFAP in the SVZ of the rats increased significantly after TBI. The positive cells began to increase at 1 day after TBI, reached the peak level at day 3 and became normal at day 14, showing significant differences between the time points of measurement following TBI and from the cell numbers in the control group measured at the same time points. The cells positive for nestin/ GFAP showed the most distinct increase in the SVZ of the rats with TBI.

CONCLUSIONTBI results in mobilization of the NSCs in the SVZ on the injured side to cause the proliferation and differentiation of the endogenous NSCs. The SVZ is one of the most important germinal centers of NSC proliferation and differentiation.

Animals ; Bromodeoxyuridine ; metabolism ; Cell Differentiation ; Cell Proliferation ; Craniocerebral Trauma ; pathology ; Glial Fibrillary Acidic Protein ; metabolism ; Lateral Ventricles ; cytology ; Nestin ; metabolism ; Neural Stem Cells ; cytology ; Neuroglia ; cytology ; Neurons ; cytology ; Phosphopyruvate Hydratase ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the relationship between the excitotoxicity and serum-inducible kinase (SNK) and spine-associated Rap GTPase-activating protein (SPAR) pathway in primary hippocampal neuron injury induced by glutamate and furthermore, to explore the molecular mechanism of neuroprotection of Zibu Piyin Recipe (ZBPYR) and the relationship between ZBPYR and the morphological regulation of dendritic spines.

METHODSThe serum containing ZBPYR was prepared by seropharmacology. Reverse transcription and polymerase chain reaction (RT-PCR) was used to detect the expression of mRNA for SNK, SPAR, postsynaptic density protein 95 (PSD-95) and N-methyl-D-aspartate (NMDA) receptor subunits (NR1, NR2A and NR2B) in primary rat hippocampal neuron cultures after pretreatment with 10 micromol/L glutamate and ZBPYR serum.

RESULTSZBPYR serum pretreatment resulted in a significant down-regulation of glutamate-induced SNK mRNA expression (P<0.05). Significant up-regulation was seen on the mRNA expression of SPAR and PSD-95 (P<0.05). All these changes were dose-dependent. The mRNA expression of NR1, NR2A and NR2B was down-regulated to different degrees (P<0.05).

CONCLUSIONThe mechanism of effect of ZBPYR on glutamate-induced excitotoxicity may be related to the regulation of SNK-SPAR signal pathway. ZBPYR may play a role in protecting and maintaining the normal morphology and structure of dendritic spines, which may be achieved by inhibiting the excessive activation of NMDA receptors.

Animals ; Cells, Cultured ; Disks Large Homolog 4 Protein ; Drugs, Chinese Herbal ; pharmacology ; GTPase-Activating Proteins ; genetics ; metabolism ; Gene Expression Regulation ; drug effects ; Glutamic Acid ; toxicity ; Hippocampus ; drug effects ; enzymology ; pathology ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Membrane Proteins ; genetics ; metabolism ; Neurons ; drug effects ; enzymology ; pathology ; Protein Kinases ; genetics ; metabolism ; Protein-Serine-Threonine Kinases ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Serum