OBJECTIVE: To observe the effect of acupotomy therapy on mRNA expressions of Bcl-2, Bax , Caspase-3 in posterior cervical extensor muscles in cervical spondylosis rabbits, and explore its mechanisms for apoptosis of cervical muscles. METHODS: New Zealand rabbits were randomly divided into normal, model, electroacupuncture (EA) and acupotomy groups, 6 in each group. The cervical spondylosis model was established by forced head-bowing for a long term. After model establishment, acupotomy was used at the trapezius muscle starting point and attachment site of sternocleidomastoid muscle, etc., once a week, total 3 times. EA was used at "Tianzhu" (BL 10), "Jingbailao" (EX-HN 15) and "Dazhu" (BL 11) for 3 weeks, 20 min a time, 3 times a week. Bcl-2, Bax, Caspase-3 mRNA expressions in posterior cervical extensor muscles were detected by real-time PCR. RESULTS: There was no significant difference in the expression of Bcl-2 mRNA among all the groups (P>0.05). Compared with the normal group, Bax mRNA increased in the model group (P<0.01), and that in the acupotomy group was lower than that in the model group (P<0.01). The ratio of Bcl-2/Bax mRNA in the model group decreased significantly compared with that in the normal group (P<0.01); in comparison with the model group, the ratio in the acupotomy group increased (P<0.01); and that in the acupotomy group was higher than the ratio in the EA group (P<0.05). The Caspase-3 mRNA expression in the model group increased compared with that in the normal group (P<0.05), and its expression in the acupotomy group decreased compared with those in the model and EA groups (P<0.05). CONCLUSIONS: Acupotomy therapy can regulate the mRNA expressions of Bax and Caspase-3, and retard apoptosis in posterior cervical extensor muscles, therefore the strained muscles are relieved, which may be one of its mechanisms for improving cervical spondylosis.

Objective To investigate the proliferation characteristics of fibroblast like synovial cells (FLS)in osteoarthritis in vitro and the mechanism of the immnnosuppressive effect of T-614 [N-(3-formy- lamino-4-oxo-6-phenoxy-4H-chromen-7-yl)methanesulfonamide ] on them.Methods FLS of OA and non- inflamed synovium(NS)were cultured and identified in vitro in the presence or absence of T-614.After incu- bation,the survival fraction(SF)of FLS was evaluated by MTT,cell cycle was observed using fluorescence - activated cell sorting(FCS)method and the expression of c-fos and COX-2 mRNA was examined by RT- PCR in FLS of OA patients.Results No statistically significant difference was noted between the OA and NS FLS in proliferation ability and cell cycle.High dose T-614 suppressed FLS SF obviously in OA and NS sta- tistically(P＜0.05),whereas the inhibition degree was not different between the two kinds of FLS.The agent induced cell apoptosis and reduced the accumulation of c-fos mRNA in OA-FLS at dose 1000 ml/L,prolonged G_1 term and shortened S term at dose 200 ml/L.The expression of COX-2 mRNA in OA FLS was suppressed obviously by T-614 at dose 1000 ml/L.Conclusion OA FLS do not display a distinct activated unlimited viability compared with NS cells,without stimulated by proinflammatory cytokine in vitro.High dose T-614 moderately inhibits the proliferation and differentiation of FLS,directly affects gene of the c-fos and COX-2 expression in OA,which may contribute to its immunosuppressive effect on OA'synovitis.

Objective To investigate the proliferative characteristics of fibroblast-like synovial cells (FLS)in osteoarthritis in vitro and the mechanism of the immunosuppressive effect of total glucusides of paeony(TGP).Methods FLS of OA and non-inflamed synovium(NS)were cultured and identified in vitro in the presence or absence of TGP.After incubation,the survival fraction(SF)of FLS was evaluated by MTI' and the TNF-?,IFN-?and bFGF level in cultured FLS supernatant was measured by ELISA.The expression of FLS c-los mRNA and cell cycle of OA-FLS was observed by RT-PCR and flow eytometry respectively at the same time.Results No statistical significant differences were noted between the OA and NS FLS in pro- liferating double time.High doses of TGP suppressed FLS-SF more evidently in OA patients than in NS(P0.05).Conclusion High dose TGP can inhibit OA-FLS proliferation,modulate cy- tokine secretion and c-fos expression in OA.This suggests that TGP has immunosuppressive effect on OA syn- ovitis,probably by preventing the synovial hypertrophy in OA.

The chemical consituents from Psychotria straminea were separated and purified by column chromatographies on silica gel, Sephadex LH-20, ODS and RP-HPLC. The strictures of the isolated compounds were identified on the basis of physicochemical properties and spectroscopic analysis, as well as comparisons with the data in literature. Sixteen compounds were isolated and elucidated as tectochrysin (1), apigenin (2), kaempferol (3), luteolin (4), diosmetin (5), quercetin (6), kaempferol-4'-O-methylether (7), rhamnetin (8), 7-hydroxycoumarin (9), 7-methoxycoumarin (10), scopoletin (11), lupeol (12), 30-oxo-lupeol (13), lupenyl acetate (14), α-amyrin (15), and ursolic acid (16). This is the first study on the chemical composition of P. Straminea, and all compounds were isolated from P. straminea for the first time. In addition to compounds 6 and 9, the other compounds were isolated from the genus Psychotria for the first time.
Coumarins ; chemistry ; isolation & purification ; Flavonoids ; chemistry ; isolation & purification ; Phytochemicals ; chemistry ; isolation & purification ; Psychotria ; chemistry ; Triterpenes ; chemistry ; isolation & purification

Objective To study the correlation between fractional anisotropy(FA)and tumor microarchitecture(MVD,VEGF and celluarity).Methods Fouteen gliomas(5 grade Ⅰ and Ⅱ,4 grade Ⅲ, 5 grade Ⅳ)confirmed histo-pathologically were performed on diffusion tensor imaging(DTI)using a GE Signa Excite Ⅱ 3.0 T MR scanner(8-channel head coil,SE echo planner imaging(EPI),thickness:5 mm, spacing:0,directions:25,B values:0 and 1000 s/mm~2,TR 6000 ms,TE minimum,FOV:240 mm? 240 mm,image matrix 128?128,NEX 2).Postprocessing was done using a DTI specific software to gain FA image.ROIs were drqwn in tumor parenchyma and the value of FA was recorded.The positive expression of VEGF and CD34 was shown using immuno-histochemistry method.The VEGF,MVD,and cellularity of every slices were recorded.Pearson correlation analysis was used.Results FA(which is 0.102?0.080 in grade Ⅰ and Ⅱ,0.171?0.037 in grade Ⅲ,0.200?0.021 in grade Ⅳ)has the trend to raise with the increasing grade of astrocytomas.FA has significant positive correlation to MVD(40/HP in grade Ⅰ and Ⅱ, 86/HP in grade Ⅲ,101/HP in grade Ⅳ),VEGF(8% in grade Ⅰ and Ⅱ,47% in grade Ⅲ,55% in grade Ⅳ),and cellularity(104/HP in grade Ⅰ and Ⅱ,160/HP in grade Ⅲ,265/HP in grade Ⅳ).The correlation coefficients between FA and VEGF,MVD,and cellularity were 0.748,0.668,0.625 respectively.Conclusion As a new imaging method,DTI can reveal the microarchitecture in gliomas and be value of distinguishing gliomas of different grade.DTI provides a new method of precise diagnosis to glioma preoperatively.

Objective To investigate of the relationship of the immunosuppression induced by Measles virus in adult patients and CD4+ CD25+ regulatory T cell. Methods Thirty-four patients with measles and 27healthy control subjects were included in this study. The whole blood was collected and CD4+ CD25+ cell andFoxP33 cell were analyzed by flow cytometry, and CD4+ CD25- and CD4+CD25+ T lymphocytes were isolated from PBMCs of patients with measles or healthy donors, CD4+ CD25- T cells were cultured in absence or presence of anti-CD3, or BCG, or live attenuated MV. The cell culture supernatant was collected after 72 hours and the concentration of IFN-γ and IL-10 was determined. Results Compared to healthy donors, we observed a reduction of the number of white blood cells and lymphocytes in patients with measles, but there was not significantly different in the frequency of CD4+ CD25+ T cells and CD4+ CD25high T cells with in the total CD4+ population in the blood. Treg from both measles patients and healthy controls significantly inhibited IFN-γ production by CD4+ CD25- T cells in response to anti-CD3 stimulation. Conclusion Induction and expansion of Treg may not represent a mechanism involved in the establishment of immune suppression by MV.

AIMTo investigate the protective effect of monosialoganglioside (GM1) on injury induced by oxygen glucose deprivation/reperfusion (OGD/Rep) in rat hippocampal slices.

METHODSThe protective effects of GM1 on hippocampal slices after OGD/Rep were observed by detecting the light transmittance (LT) changes of rat hippocampal slices and 2, 3, 5-triphenyltetrazolium chloride (TTC) staining of rat hippocampal slices.

RESULTS(1) In four groups treated with 0 (control), 0.1, 1.0, 10 micromol/L GM1, the peak of light transmittance (LT) in the slices treated with 1.0 micromol/L GM1 was significantly lower than that of the control and the group treated with 0.10 micromol/L GM1 (P < 0.01, ANOVA), while the peak of LT in the slices treated with 10.0 micromol/L GM1 was significantly lower than that of the other groups (P < 0.01, ANOVA). The time to reach the peak of LT in four groups was significantly different from each other (P < 0.05, Kruskal-Wallis test). The time to reach the peak of LT in the group treated with 1 micromol/L GM1 was the significantly longer than that in the control (P < 0.01, Mann-Whitney U test). (2) There was characteristic dose-response relationship between GM1 and TTC staining of rat hippocampal slices. In the five groups, treated with 0 (control), 0.01, 0.1, 1.0, 10 micromol/L GM1 respectively, TTC staining in the group treated with 1 micromol/L GM1 was the deepest (P < 0.05 vs. control, 0.01 and 0.1 micromol/L GM1 group, ANOVA), and the next was in the group treated with 10 micromol/L GM1 (P < 0.05 vs. control and 0.01 micromol/L GM1 group, ANOVA).

CONCLUSIONGM1 could protect injury induced by OGD/Rep in rat hippocampal slices effectively in vitro.

Animals ; G(M1) Ganglioside ; pharmacology ; Glucose ; deficiency ; Hippocampus ; metabolism ; Hypoxia ; metabolism ; In Vitro Techniques ; Male ; Oxygen ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury

Based on standard carotid endarterectomy, we performed modified carotid endarterectomy in two cases of carotid artery stenosis by changing the direction of the carotid artery incision to avoid restenosis of the internal carotid artery without using a patch. The two patients recovered smoothly without any complications. Compared with eversion or patch endarterectomy, this modified carotid endarterectomy avoids restenosis of the carotid artery and shortens operation time.

Gastrodia elata Bl. is a famous and costful traditional Chinese medicine. Their genomic DNA fingerprints were investigated using a modified Randomly Amplified Polymorphic DNA method. DNA fragments common to all or to fine populations were identified and recovered. Five DNA fragments were proven not to be reported through DNA cloning, PCR identifying, nucleotide sequencing and bioinformatics analyses and were received in and recorded by NCBI GenBank. Gastrodine contents of the Gastrodia tuber samples were determined using high performance liquid chromatography technique. The distribution of the five DNA fragments in 9 Gastrodia elata Blue populations and the correlation with gastromedicine content were studied. The results show the distribution of these DNA sequences varied greatly among the populations whereby DNA Sequence 1 was the common and distinguishing molecular marker for all the populations studied and DNA Sequence 2 may relate to higher gastrodine content. In conclusion, these DNA marker sequences can be employed to identify genuine gastrodia tubers, better varieties and optimize their selection and cultivating.
Base Sequence ; Benzyl Alcohols ; analysis ; Cloning, Molecular ; Computational Biology ; DNA, Plant ; chemistry ; Gastrodia ; genetics ; Glucosides ; analysis ; Plant Tubers ; genetics

OBJECTIVETo elucidate the variation in characterizations and genetic evolution of the matrix protein 2 or ion channel protein(M2) genes of avian influenza subtype H5N1 viruses in the boundary region of Yunnan province from 2008 to 2012.

METHODSA total of swab samples were collected from foreign poultry such as the junction between Yunnan and Vietnam, Laos,myanmar and wild birds in boundary region of Yunnan province from 2008 to 2012 and screened by H5N1 subtype-specific multiplex RT-PCR. The M genes of H5N1 virus from the positive samples were amplified by RT-PCR and cloned into pMD18-T vectors for sequencing. The alignment and phylogenetic analysis of M2 genes were performed with sequences of the known reference strains.

RESULTSA total of 71 positive samples were found out of 1240 samples and the positive rate was 5.72%. A total of 14 different M2 sequences were obtained from 30 positive samples and were divided into 3 distinct clades or sub-clades(1.2.1, 1.2.2 and 2) by phylogenetic analysis, 5, 7 and 2, respectively. The M2 genes and Hemagglutinin(HA) genes of H5N1 viruses from the boundary region of Yunnan province had showed different relationship of genetic evolution. The substitution or mutation of key amino acids sites had been found among the domains of epitope, adamantane-resistance, and poultry or human original viral strains.

CONCLUSIONThe M2 genes of H5N1 subtype viruses in boundary region of Yunnan province from 2008 to 2012 showed genetic divergence and the virus of clade 1.2.2 had become dominant epidemic strain in this region.

Animals ; Birds ; virology ; Chickens ; virology ; China ; Evolution, Molecular ; Influenza A Virus, H5N1 Subtype ; classification ; genetics ; Influenza in Birds ; virology ; Phylogeny ; Poultry ; virology ; Viral Matrix Proteins ; genetics