The Chinese acupuncture doesn't only belong to China but rather to the world.The boom of overseas acupuncture education and scientific research as well as the satisfactory working environment compels us to reflect the Chinese traditional medicine education in our country.Especially in higher education specialty courses of acupuncture should not be subdivided;meanwhile,much attention should be paid to elementary education of western medicine.

Objective To study the anti-tumor effect of 5-aza-2′-deoxycytidine(5-aza-dC) on breast carcinoma xenografted in nude mice. Methods The model of breast carcinoma xenografted in nude mice was established.Ten mice were randomized into the treatment group(treated with 5-aza-dC) and control group(treated with PBS).The mass of the tumors before and after treatment were measured in the two groups,and the inhibition rate of the tumor was calculated and the growth curve was drawn.Immunohistochemistry and Western blotting were employed to detect the expression of normal epithelial cell specific-1(NES1)gene. Results The inhibition rate of the tumor in the treatment group was 57.44%,which was significantly different from the control group(P

OBJECTIVE Apolyclonalantibody-basedhomogeneouschemiluminescenceimmunoas-say was developed and optimized using AlphaLISA technology for the quantitative detection of microcys-tin-LR(MC-LR)inwatersamples.METHODS Thismethodwasbasedonacompetitivemodelin which an immune complex was formed from the ingegral binding of artificial MC-LR antigen-coated lumi-nescene beads,free MC-LR standards or sa mples,antibody and biotinylated second antibody.Next sensor bead were added that approached the i mmune co mplex through biotin-streptavidin interaction. With the exciting light,the energy was passed from the sensor luminescer before a special emission light could be observed.To opti mize the reaction conditions,working dilutions of polyclonal antibody and bioti-nylated second antibody were assayed while the effect of buffer syste ms and ti me of each reaction were evaluated.RESULTS Maininfluencingfactorsoftheassaywerediscussedasworkingdilutionsofpoly-clonal antibody and biotinylated goat anti rabbit IgG,assay buffer and reacting ti me.After opti mization of reaction conditions,MC-LR AlphaLISA could be finished in 40 min,with a sensitivity of 0.006 μg·L-1 and a dynamic range of 0.006 -5 μg·L-1 .The coefficient of variation was below 10% and average recovery was 1 07.7%.Moreover,the cross reactivity rates of MC-RR and MC-RY to MC-LR were 13.2%and0.91%,respectively.CONCLUSION Thismethodishighlysensitiveandspecific,time-saving and quite suitable for high throughput determination of MC-LR water samples.

Objective To investigate the impact of isoniazid (INH)-resistant Mycobacterium tuberculosis (Mtb) on the prevalence and dissemination of multi-drug resistant tuberculosis (MDR-TB).Methods A total of 251 patients diagnosed with tuberculosis in designated hospitals of Guanyun,Jiangsu and Deqing,Zhejiang from 2010 to 2011 were included in the study.The drug susceptibility tests (DST) were performed on all the Mtb isolates available from the sputum cultures.Mycobacteral interspersed repetitive units-variable number tandem repeats (MIRU-VNTR) was conducted for genotyping for all available Mtb isolates.Chi-square test,Fisher exact test,ANOVA and non-conditional Logistic regression modelling were applied for data analysis.Results Among 251 patients with Mtb isolates and DST results available,72 (28.7％) were resistant to INH,including 13 were INH mono-drug resistant.Of the remaining 59 INH-resistant Mtb,34 (13.5％) were resistant to rifampin TB and 25 were resistant to streptomycin and/or ethambutol.The clustering analysis based on MIRU-VNTR genotyping revealed 29 clustered genotypes (including 105 isolates) and 146 unique genotypes (including 119 isolates).Twentyfive clusters contained drug resistant Mtb and 16 clusters of them comprised by 37 INH-resistant isolates and 20 MDR-TB isolates,which accounted for 51.4％ of the INH-resistant isolates and 58.8％ of the MDR-TB isolates.Single factor analysis showed that sex,age,previous tuberculosis treatment history and sputum smear results were all related to INH-resistant tuberculosis and MDR-TB (all P ＜ 0.05).Multiple factors analysis showed that previous tuberculosis treatment history was risk factor of MDR-TB (OR=8.40,95 ％CI:3.342-21.105),while the risk factors of INH-resistant tuberculosis were previous tuberculosis treatment history (OR=3.52,95％CI:1.570-7.910),pulmonary caviry (OR=2.27,95％CI:1.075-4.799) and sputum smear results (OR=0.50,95％CI:0.275-0.892).Conclusions That INH-resistant strain may evolve to the MDR-TB after recent transmission is a possible trend.Patients with previous treatment history and advanced age are at high risk of INH-resistant tuberculosis and MDR-TB.

The zinc-regulated transporters (ZRT), iron-regulated transporter (IRT)-like protein (ZIP) plays an important role in the growth and development of plant. In this study, a full length cDNA of ZIP encoding gene, designed as DoZIP1 (GenBank accession KJ946203), was identified from Dendrobium officinale using RT-PCR and RACE. Bioinformatics analysis showed that DoZIP1 consisted of a 1,056 bp open reading frame (ORF) encoded a 351-aa protein with a molecular weight of 37.57 kDa and an isoelectric point (pI) of 6.09. The deduced DoZIP1 protein contained the conserved ZIP domain, and its secondary structure was composed of 50.71% alpha helix, 11.11% extended strand, 36.18% random coil, and beta turn 1.99%. DoZIP1 protein exhibited a signal peptide and eight transmembrane domains, presumably locating in cell membrane. The amino acid sequence had high homology with ZIP proteins from Arabidopsis, alfalfa and rice. A phylogenetic tree analysis demonstrated that DoZIP1 was closely related to AtZIP10 and OsZIP3, and they were clustered into one clade. Real time quantitative PCR analysis demonstrated that the transcription level of DoZIP1 in D. officinale roots was the highest (4.19 fold higher than that of stems), followed by that of leaves (1.12 fold). Molecular characters of DoZIP1 will be useful for further functional determination of the gene involving in the growth and development of D. officinale.
Amino Acid Sequence ; Cloning, Molecular ; Dendrobium ; chemistry ; classification ; genetics ; metabolism ; Gene Expression Regulation, Plant ; Iron ; metabolism ; Membrane Transport Proteins ; chemistry ; genetics ; metabolism ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Plants ; chemistry ; classification ; genetics ; Sequence Alignment ; Zinc ; metabolism

Objective To compare and analyze the phylogenetic tree and sequence variant characteristics of Klebsiella species between 16S rDNA and rpoB. Methods Eighteen isolates identified as genus Klebsiella (with 15 of K. Pneumoniae and 3 of K. Oxytoca) by automated biochemical tests were selected. DNA were extracted, 16S rDNA and rpoB genes were amplified and sequenced with Klebsiella 16S rDNA and rpoB primers. Together with already published 8 species of Klebsiella and 9 species of Enterobacteriaceae 16S rDNA and rpoB sequences from GenBank, totally 35 sequences of 16S rDNA and rpoB respectively, phylogenetic trees were constructed with MEGA 4.0 to the analysis of groups. DNAStar/MegAlign was used for comparison of variable regions of 16S rDNA and rpoB, with analysis of degree of divergent at the same time. Results As for all 35 sequences, both 16S rDNA and rpoB phylogenetic trees divided Klebsiella species into three groups, 15 of K. Pneumoniae in this study and 6 of K. Pneumoniae from GenBank (except for K. Oxytoca and K. Mobilis) cluster to group Ⅰ, K. Oxytoca and K. Mobilis were cluster to group Ⅱ and Ⅲ, respectively. In rpoB phylogenetic tree, no matter group Ⅰ and group Ⅱ, or subgroup within group Ⅰ, the bootstrap values in each node of rpoB phylogenetic tree is obviously higher than that of 16S rDNA. Moreover, as for cluster to K. Oxytoca, rpoB is better than 16S rDNA. Analysis nucleic acid sequences of Klebsiella species, with 41 variable regions and 4 most significant regions were found within the Klebsiella 16S rDNA, while rpoB with 63 variable regions, and 1 most significant region. The similarity of 16S rDNA and rpoB within Klebsiella were 95.9%-100% and 90.2%-100% respectively. Further analysis divergent degree of 16S rDNA and rpoB within Klebsiella, the divergent value of rpoB (0-10.6) is higher than that of the 16S rDNA(0-4.0). Conclusion As for molecular classification and identification within KlebsieUa species, rpoB has more advantages than 16S rDNA.

Objective Research on traumatic the related factors of sleep disorder after traumatic brain injury ,in order to pro-vided the rationale for the diagnosis and treatment .Methods The SPIEGEL was used to evaluate the traumatic brain injury patients who were hospitalized .Recording time in sleep disorders in 3 months .Analysis the relations between the sleep disorders and brain injury site by combining with the patients head CT and MRI .Results Seen in 200 cases of patients with sleep disorders of 105 cases (52 .5% );71 cases appeared in patients within 1 week after waking ,accounted for 76 .19% ;The brain stem ,frontal lobe and basal ganglia injury occurred sleep disorders were more likely(66 .7% ,64 .0% ,70% ) .The difference was statistically significant(P<0 .05) .Conclusion Sleep disorder is a common clinical symptom of mild traumatic brain injury .a time to focus on the patients with-in 1 week after waking ,and closely related to brain stem ,frontal lobe and basal ganglia injury .

A dual-label time-resolved fluoroimmunoassay was established for simultaneously detecting pepsinogen Ⅰ (PGⅠ) and pepsinogen Ⅱ (PG Ⅱ) in human serum. Two capture monoclonal antibodies, 8003# of PGⅠ and 8101# of PGⅡ, were co-coated in 96 microtitration wells. The counterpart tracer monoclonal antibodies, 8016# of PGⅠ and 8102# of PGⅡ, were labeled with Eu3+ and sm3+-chelates, respectively. The samples were assayed by one-step sandwich protocol with the time-resolved fluorometry. The measurement ranges of PGⅠ were 0.2～300.0 ?g/L with the within-run and between-run precision was 5.2% and 8.1%, and that of PGⅡ were 0.05～55.0 ?g/L with the within-run and between-run precision was 7.1% and 11.7%, respectively. The average recovery rates of PGⅠ and PGⅡ were 96.9% and 103.7%, respectively. The results obtained by the dual-label assay agreed well with those by enzyme-linked immunosorbent assays of PGⅠ and PGⅡ, whose correlation ratio were 0.9426 of PGⅠ and 0.9396 of PGⅡ, respectively. The means of 300 healthy volunteers were (157.3 ? 51.0) ?g/L for serum PGⅠ,(10.6 ? 5.9) ?g/L for serum PGⅡ, and (14.8 ? 4.3) for the PGⅠ/PGⅡ ratio. The normal ranges of serum PGⅠ levels for healthy volunteers were 55.3～259.3 ?g/L, those of serum PGⅡ levels were less than 23 ?g/L, the PGⅠ/PGⅡ ratio was more than 6. The proposed dual-label TRFIA for simultaneous detection of PGⅠ and PGⅡ is a simple, sensitive, and rapid method. It could provide serology high-screening of the samples for gastric diseases and would allow investigations into the possible diagnostic value of analysis in various clinical condition.

Objective To establish a time resolved fluoroimmunoassay (TRFIA) method for detecting Glypican 3 (GPC3) and to explore the diagnostic value of serum GPC3 for hepatic carcinoma (HCC). Methods Microplate coated with anti-GPC3 monoclonal antibody 7C8 and GP9 labeled with Eu3+ were used to establish TRFIA kit. The serum concentrations of GPC3 in 41 HCC patients and 44 chronic hepatitis (CH) patients were quantitatively analyzed. AFP was detected by with lowest limit of 2.06 μg/L. The CV of inter and intra assay were 12.25% and 12.91%, respectively. The average serum concentration of GPC3 in HCC patients was (86.68±110.39) μg/L (median: 56.98 μg/L). But in CH patients it was only (14.77±29.48) μg/L, which was significantly lower than that in HCC (Wilcoxon W=1335.00, Z=-4.99, P<0.001). With diagnostic cut-off value set at 42.94 μg/L, the diagnostic sensitivity and specificity of TRFIA GPC3 for HCC were 58.5% (24/41) and 95.5%(42/44) respectively. The diagnostic sensitivity of AFP was 46.3% (19/41) in 41 HCC patients, and was raised to 78.0% (32/41) when combined with GPC3. Conclusions Serum GPC3 assay by TRFIA is established and it could increase the diagnostic sensitivity for HCC when combined with AFP.

Objective To study the effect of chest wall vibration therapy on bronchiolitis. Methods A total of 64 patients with bronchiolitis were divided into an experimental group and a control group, the former included 34 cases and the latter included 30 cases. The experimental group received both routine treatment and chest wall vi- bration, while the control group only received routine treatment. PaO_2, PaCO_2, SaO_2, Heart Rate (HR) and Respi- ration (R) were observed, respectively, in the experimental group and the control group at the beginning and the end of the third day. Time needed for expectoration and length of hospital stay in the two groups were observed. Results It was shown that PaO_2, PaCO_2, SaO_2 , HR, R were significantly improved at the end of the third day when compared with those at the beginning in both groups(P