Using the immunohistochemical method combined with tract tracing techniques, a new subdivision, the marginal division, was found in the striatum of the rat. In this subdivision, substance P, enkephaline and dynorphin B terminals were densely localized. The present study investigated the synaptic organization of the substance P-like immunoreactive nerve terminals in the marginal division. Four major types of substance P immunoreactive synapses were identified. Axodendritic and axospinous synapses were the most common ones. Compound synapses with more than two synaptic components were also observed. One axo-axonic synapse was present. Both symmetric and asymmetric substance P-like immunoreactive synapses were seen in the marginal division. The existence of asymmetric synapses, small postsynaptic dendrites and axo-axonic synapses, distinguished the substance P-like immunoreactive nerve terminals in the marginal division from those in the other part of the striatum. The characteristics and complexity of the synaptic organization suggested that the substance P-like immunoreactive nerve terminals in the marginal division probably orginate from a different source and have a different function, as compared to those in the rest part of striatum.

Objective:To study the isolation, culture and identification of the TMJ cells and to observe the biological characteristics of cultured fibrochondrocytes. Methods:The TMJ discs were dissected from two 1 month goats under sterile conditions and were digested with collagenase. The cells were collected. Morphological changes and attachment efficiency were constantly observed under phase-contrast microscope. Immunohistochemical staining for type I collagen as well as toluidine blue staining were performed. Ultrastructures of the TMJ cells were observed under transmission electron microscope. Results: Most of the primary fibrochondrocytes presented a short spindle-shape while the rest showed polygon-shape. On the 7th day, the perliferating fibrochondrocytes started to contact each other to form a monolayer covering the bottom of the incubation disc. Immunohistochemical staining of type I and toluidine blue staining exhibited positive results. The fibrochondrocytes cytoplasms were rich in mictochondria and endoplasm reticulum. Conclusion: The fibrochondrcytes isolated from one-month-old goat TMJ disc have good proliferation ability in vitro and cells from passage 1 to 3 might be used as seed cells for TMJ disc tissue engineering.

BACKGROUND:There is no common cognition in the cell type in the temporomandibular joint(TMJ)discs,and names describing TMJ disc cells also vary a lot.OBJECTIVE:To characterize the type and the distribution of cells in the TMJ disc of goats DESIGN,TIME AND SETTING:A single sample observation was completed in Cettutar and Motecutar Biologicat Center and Electron Microscope Center of Lanzhou University from March to May in 2007.MATERlALS:TMJ discs were obtained from two one-month-old healthy goats that were slaughtered freshly.METHODS:Bilateral TMJ discs of goats were cut off completely and were divided into 6 parts by 3 cuts in the major axis direction (mediolaterally)and 2 cuts in the minor axis direction(anteroposteriorly).Then the marked samples were fixed in 10%neutral formalin Iiquid for 24 hours and embedded by paraffin.MAIN OUTCOME MEASURES:Hematoxylin and eosin staining were used to identify regional variation of cell type and cellnumbers.Toluidine blue staining and collagen type Ⅰimmunohistochemical assay were performed to test the distribution of collagens.Transmission etectren microscopy was used to observe the ultrastructure of cells of goat TMJ discs.RESULTS:TMJ discs were comprised of cells and collagen fibers distributing unevenly.Collagens were mostly type Ⅰ.Collagen fibers were wave or crimping and approximately parallel to each other.with cells scattered in their matrix.Fibroblast-like cells and chondrocyte-like cells were the main two types of cells existing,with the former predominating over the later in a ratio of 2.05:1 approximately.There were no significant regional differences in cell type and distribution statistically.Transmission electron microscopy denoted that fibroblast-iike cells have fairly larger fusiform or irregular nuclei with very few organelles,while the chondrocyte-like cells exhibited round or elliptical nuclei,well defined pericellular electron lucent zones,unconspicuous cytocysta and non-distinctive pseudopodia CONCLUSION:There are no significant differences in type,number and arrangement of cells in TMJ discs of one-month-old goats statistically,with Fibroblast-like cells predominating slightly over chondrocyte-like cells.

Objectives To investigate the effect of magnetron sputtered niobium nitride (NbN) on the bonding strength of commercially pure cast titanium (Ti) and low-fusing porcelain (Ti/ Vita titankeramik system).Methods Sixty Ti specimens were randomly divided into four groups, group Tl, T2, T3 and T4.All specimens of group Tl and T2 were first treated with 120 μm blasted Al2O3 particles, and then only specimens of group T2 were treated with magnetron sputtered NbN film.All specimens of group T3 and T4 were first treated with magnetron sputtered NbN film and then only specimens of group T4 were treated with 120 μm blasted Al2O3 particles.The composition of the deposits were analyzed by X-ray diffraction (XRD).A universal testing machine was used to perform the three-point bending test to evaluate the bonding strength of Ti and porcelain.The microstructure of NbN, the interface of Ti-porcelain and the fractured Ti surface were observed with scanning electron microscopy (SEM) and energy depressive spectrum (EDS) , and the results were compared.Results The XRD results showed that the NbN deposits were cubic crystalline phases.The bonding strength of Ti and porcelain in Tl to T4 group were (27.2±0.8) , (43.1±0.6) , (31.4±1.0) and (44.9±0.6) MPa.These results were analyzed by one-way analysis of variance and differences between groups were compared using least significant difference test Significant inter-group differences were found among all groups (P<0.05).The results of SEM showed that with treatment of Al2O3 or NbN, alone, pre-cracks were found in the interface of Ti-porcelain, while samples treated with both Al2O3 and NbN had better bond.EDS of Ti-porcelain interface showed oxidation occurred in T1, T2 and T3, but was well controlled in T4.Conclusions Magnetron sputtered NbN can prevent Ti from being oxidized, and can improve the bonding strength of Ti/Vita titankeramik system.Al2O3 blast can also improve the bonding strength of Ti/Vita titankeramik system.

OBJECTIVETo investigate the effect of component I from agkistrodon acutus venomon (AAVC-I) the migration of human umbilical vein endothelial cells (HUVECs), and to elucidate the possible anti-angiogenic mechanism of AAVC-I.

METHODSThe effect of AAVC-I on the migration of HUVECs which was cultivated in vitro and treated with AAVC-1 at four concentrations: 0, 20, 40, 80 microg/ml, was observed by methods of scratch wound-healing and Transwell assay. The expression level of mRNA and protein of P-selectin and intercellular cell adhension molecule-I (ICAM-1) were examined by RT-PCR and Western blot assay.

RESULTSCompared with the blank group, the migration ability of HUVECs in each AAVE-I treated group was reduced in a dose-dependent manner, and the expression level of the mRNA and protein of P-selectin and ICAM-1 were decreased.

CONCLUSIONAAVC-I inhibits the migration of endothelial cell, which is acted by down-regulation of the expression content of mRNA and protein of P-selectin and ICAM-1.

Cell Movement ; drug effects ; Cells, Cultured ; Crotalid Venoms ; pharmacology ; Down-Regulation ; Human Umbilical Vein Endothelial Cells ; drug effects ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; P-Selectin ; metabolism ; RNA, Messenger

To improve ethanol production in Saccharomyces cerevisiae,an integration plasmid pUPGKAT with PGK promoter(phosphoglycerate kinase promoter),adh1 gene(the coding sequences of alcohol dehydrogenaseⅠ) and CYC1 terminator(Cytochrome c transcription terminator) was constructed.Firstly,a fusion fragment composed of PGK promoter and adh1 gene was generated by over lap extension PCR and ligated into pUG6 resulting in plasmid pUPGKA.Subsequently,CYC1 termi nator was amplified from pSH65 by PCR and ligated to the SpeⅠand SacⅡrestriction site of pUPGKA.To integrate PGK-adh1-CYC1 into S.cerevisiae genome,pUPGKAT was digested by TthⅢⅠand the lin-earized plasmid was used to transform S.cerevisiae YS2-△adh2(adh2 disrupted strain) by lithium acetate method.The yeast mutant YS2-△adh2-adh1 which had the adh1 gene placed under the PGK promoter and harbored the adh2 deletion was constructed.Anaerobic fermentation showed overexpression of adh1 by PGK promoter resulted in a 8.84% higher ethanol production compared to YS2-△adh2.

Acyl carrier protein is an essential component involved in the biosynthesis of DHA(Docosahexaenoic Acid) via PKS(Polyketide synthase) pathway,which takes the growing acyl chain from one enzyme to another.One cDNA clone,with high homology of ACP,was isolated from Schizochytrium sp.FJU-512 cDNA library.The deduced amino acid sequence contained 142 residues with isoelectric point of 5.04 and had the 4'-phosphopantetheine prosthetic(4'-PP) binding site.The target fragment was digested with BamHⅠ/HindⅢand inserted into the expression vector pET-30a resulting in the plasmid pET-30a/acp.The recombinant vector was transformed into E.coli BL21(DE3) and induced by IPTG.SDS-PAGE analysis demonstrated that ACP was effectively expressed.

The influence factors for hydrogen production using photosynthetic bacteria group were studied. The effects of carbon sources, nitrogen sources, carbon concentration, nitrogen concentration, initial pH value, illumination mode and inoculation amount on hydrogen production were investigated by experiment of single factor and orthogonal experiment. The optimum process conditions were as follows: the best bacteria group was number 3, the glucose was carbon source and concentration of carbon source was 3 g/L; the urea was nitrogen source and concentration of nitrogen source was 9 g/L; the inoculation amount was 10%; initial pH value was 8.5; the illumination mode was 12 h light and 12 h dark alternation; incubation temperature was 30?C. The main factors affecting hydrogen production were strain, carbon sources, carbon source concentration and nitrogen suorces.

Objective To investigate the feasibility of ~1H-MR spectroscopy(~1H-MRS)imaging to quantitatively detect fatty liver.Methods Twenty patients with fatty liver and 11 healthy volunteers underwent plain CT scan,conventional MR imaging and ~1H-MRS analysis.The blood lipid and liver function were tested on the same day as the MR examination.~1H-MRS sequence measured the peaks of H_2O and lipid,and the areas under the peaks.The relative contents of the lipid compound were calculated,and compared with the results of CT scan and liver function tests.Results The CT values of the normal group and the fatty liver group were(59?9)HU and(24?11)HU respectively.On ~1H-MRS a protruding high H_2O peak and a flat low lipid peak were observed in the normal group,while the protruding high H_2O peak and a high lipid peak appeared in the fatty liver group.The values of lipid peak in the normal group and the fatty liver group were(0.05?0.01)?10~5,(0.70?0.24)?10~5 respectively(t=4.32,P0.05),the areas under the lipid peak were(1.36?0.73)?10~9、(2.35?1.15)?10~9 respectively(t=5.21,P0.05).Conclusion ~1 H-MRS imaging is feasible to quantitatively detect liver fat and is a non-invasive method for detecting early fatty liver.

Objective To construct the recombinant plasmid pYES6-S and express and purify spike protein of severe acute respiratory syndrome(SANS)coronavirus in Saccharomyces cerevisiae. Methods DNA fragments of SANS coronavirus were obtained by reverse transeription.Four over- lapped fragments of spike protein genes were amplified by polymerase chain reaction(PCR)and ligated into an integral spike protein gene by restriction enzyme digestion.The spike protein gene recombined with pYES6 and cloned into E.coll.The recombinant plasmid pYES6-S was induced and expressed in Saccharomyces cerevisiae(INVScl)by galactose.Results The recombinant plasmid pYES6-S was confirmed that inserted fragment was right in length,direction and base matching by restriction enzyme digestion and sequencing.The purified protein encoded by the whole spike protein gene was about Mr 110?10~3 identified by electrophoresis.Conclusion The whole spike protein gene of SARS coronavirus is cloned into E.coli and the protein is expressed in Saccharomyces cerevisiae successful ly.which can be helpful in SARS vaccine research.