Objective To discuss the influence and significance of stromal cell-derived factor 1 (SDF-1) and its specific receptor CXC chemokine receptor 4 (CXCR4) in proliferation, migration and invasion ability of SW480 colorectal cancer cells. Methods The colorectal cancer cell line SW480 in logarithmic phase was divided into four groups:control group (with no any processing), SDF-1 group (added 100μg/L SDF-1), SDF-1+1 mg/L AMD3100 mixed group (added 1 mg/L AMD3100 for 2 hours, then added 100μg/L SDF-1) and AMD3100 group (added 1 mg/L AMD3100). Immunohisto?chemistry method was used to detect the protein expression of CXCR4 in SW480 cells. The expression of CXCR4 mRNA in SW480 cells was detected by RT-PCR before and after SDF-1 and AMD3100 treatment. MTT assay and transwell chamber were used to test the changes of proliferation, migration and invasion ability of SW480 cells before and after SDF-1 and AMD3100 treatment. Results The result of immunohistochemistry showed that CXCR4 protein was expressed in SW480 cells (positive rate=80%). CXCR4 mRNA was expressed in SW480 cells. The expression of CXCR4 mRNA was up-regulat?ed by SDF-1(100μg/L), which could be inhibited by AMD3100 (1 mg/L). The proliferation activity was higher in SDF-1 group (0.847±0.039) compared to that in control group (0.624±0.011) and SDF-1+AMD3100 mixed group (0.607 ±0.016). The proliferation activity was lower in AMD3100 group (0.456 ± 0.031) than that in control group and SDF-1+AMD3100 mixed group (F=108.03, P<0.05). The number of transmembrane cells was more in SDF-1 group (98.7±5.8, 33.7±6.2) than that in control group (21.0±2.2, 6.1±2.3), SDF-1+1 mg/L AMD3100 mixed group (18.5±8.4, 8.5±2.8) and AMD3100 group (12.1±3.2, 2.1±1.0) detected by transwell chamber experiment. However, there were no statistical differences between three groups. Conclusion The biological axis SDF-1/CXCR4 can promote the proliferation, migration and invasion in colorectal cancer cell line SW480.

Objective To determine the median effective concentration (EC50) of remifentanil inhibiting responses to tracheal extubation in patients of Uygur nationality.Methods The patients of Uygur nationality,aged 18-60 yr,of ASA physical status Ⅰ or [Ⅱ,scheduled for elective surgery,were enrolled in this study.After the end of surgery,remifentanil was given by target-controlled infusion until tracheal extubation.The initial target plasma concentration (Cp) of remifentanil was 2.0 ng/ml.The response was defined as positive when MAP and/or HR increased by ＞ 15％ of the level at the end of operation and the duration ＞ 15 s during extubation.Each time the target Cp increased/decreased if the cardiovascular response was positive or negative.The ratio between the two successive concentrations was 1.1.The EC50 and 95 ％ confidence interval of remifentanil blunting responses to extubation was calculated by Probit method.Results Twenty-eight patients completed the study.The EC50 and 95 ％ confidence interval of remifentanil required for inhibiting the responses to extubation was 1.75 ng/ml and 1.45-2.01 ng/ml.Conclusion The EC50 of remifentanil inhibiting the responses to tracheal extubation is 1.75 ng/ ml in patients of Uygur nationality.

To study the bioactive polypeptides included in Bufo skin and its secretions the plasmid skin cDNA library of adult Japanese toad Bufo japonicus formosus was prepared. The pSD64TR has been used as the vector and the cloning sites are Xho I and EcoR I. To screen cDNAs encoding bioactive components, the plasmid cDNA library was transformed into E. coli DH5 competent cells, and positive colonies were screened by colony PCR (polymerase chain reaction). The suspension of a single colony in LB medium was used as the template, SP6 (the upstream primer of the plasmid cDNA library) and a primer with Xho I site and polyT were used as the primers. As the result, 465 positive colonies out of 1 344 were obtained and their plasmid were collected and sequenced. By homologous analysis, it was found that one of the cDNAs encoding a peptide with high homolog with transgelin-2, which was registered in GenBank (accession number: JX197456), and it was indicated as a partial cDNA sequence with a deletion at the 5' end. The transcript is 997 bp consisting of 31 bp 5', 618 bp 3' untranslated region (UTR) and an open reading frame (ORF) of 348 bp encoding a polypeptide of 115 amino acids. In the putative protein product, there is a calponin homology domain, two cysteine residues for a disulfide bond and three a-helix domains, and five potential phosphorylation sites. The homologous analysis indicates 90% similarity with Xenopus (Silurana) tropicalis and 89% with Xenopus laevis, and 71%-85% with other species.

Objective To analyze the reasons of focal liver lesions that difficult to detect by conventional ultrasound ultrasound-CT/MR fusion imaging.Methods 101 lesions which were confirmed by pathology or clinical diagnosis standards were recruited in the research.All of them were difficult to detect by conventional ultrasound but CT/MR display clearly.Ultrasound-CT/MR fusion imaging was used to observe the size,location and internal echo of the lesions,as well as the background of the surrounding liver parenchyma.Results All cases were successfully registrated,the registration time were 2-6min [(4.1 ±0.6)min].For these 101 lesions,93.1％(94/101) of which the diameter ≤20 mm,56.4％ (57/101) were located in hepatic segments near the diaphragm (such as S2,S4,S7,S8),78.2％ (79/101) were internal isoecho,and 79.2％(80/101) in the background of liver cirrhosis.Conclusions The important reasons that focal liver lesions detected difficult by conventional ultrasound includes:lesion size,location,internal echo and the hepatic background.

This paper discusses application of multimedia and network technology in Laboratory Animal Science Teaching:We have set up multimedia courseware,made animal experiment video and built network course-learning system and a good teaching effect has been achieved.Then it analyzes the problems of multimedia and network technology in Laboratory Animal Science Teaching.

OBJECTIVEStudy on correlation between H2O2 scavenging system and flavonoids accumulation of Scutellaria baicalensis.

METHODThe content of baicalin and baicalein in suspension cell of S. baicalensis was determined by HPLC. The content of total flavonoids and H2O2, the activity of POD and PAL was detected by UV spectrophotometry.

RESULTThe content of total flavonoid and the activity of PAL increased significantly in 12 days after 40 degrees C, dark and PEG stress. Around 12 days after NPA, NPA +40 degrees C, 40 degrees C, NPA + dark, dark and PEG stress, the content of baicalin declined and the content of baicalein rise, the activity of POD showed an increasing trend, and level of H2O2 remain stable.

CONCLUSIONModerate environmental stress could promote the accumulation of total flavonoids in S. baicalensis, baicalin convert to baicalein by POD, and maintaining the stability of H2O2 content to avoid oxidative damage.

Anti-Infective Agents ; analysis ; metabolism ; Antioxidants ; analysis ; metabolism ; Cells, Cultured ; Darkness ; Flavanones ; analysis ; metabolism ; Flavonoids ; analysis ; metabolism ; Hot Temperature ; Hydrogen Peroxide ; analysis ; metabolism ; Oxidants ; metabolism ; Oxidative Stress ; Peroxidase ; metabolism ; Phenylalanine Ammonia-Lyase ; metabolism ; Plant Extracts ; chemistry ; Plant Roots ; chemistry ; drug effects ; enzymology ; metabolism ; Plants, Medicinal ; Polyethylene Glycols ; pharmacology ; Scutellaria baicalensis ; chemistry ; drug effects ; enzymology ; metabolism ; Stress, Physiological

Single nucleotide polymorphisms (SNP) is an important molecular marker in traditional Chinese medicine research, and it is widely used in TCM authentication. The present study created a new genotyping method by combining restriction endonuclease digesting with melting curve analysis, which is a stable, rapid and easy doing SNP genotyping method. The new method analyzed SNP genotyping of two chloroplast SNP which was located in or out of the endonuclease recognition site, the results showed that when attaching a 14 bp GC-clamp (cggcgggagggcgg) to 5' end of the primer and selecting suited endonuclease to digest the amplification products, the melting curve of Lonicera japonica and Atractylodes macrocephala were all of double peaks and the adulterants Shan-yin-hua and A. lancea were of single peaks. The results indicated that the method had good stability and reproducibility for identifying authentic medicines from its adulterants. It is a potential SNP genotyping method and named restriction endonuclease digest - melting curve analysis.

A FatB unigene was obtained from the transcriptome dataset of Lonicera japonica Thunb. Full-length FatB cDNA was cloned from buds of Lonicera japonica Thunb., Lonicera japonica Thunb. var. chinensis (Wats.) Bak., Lonicera hypoglauca Miq. and Lonicera dasystyla Rehd. using RT-PCR technology, and named as LJFatB, LHFatB, LJCFatB and LDFatB. The results of bioinformatic analysis showed that LJFatB, LJCFatB, LHFatB and LDFatB and Arabidopsis thaliana AtFatB had a closely relationship. Nucleotide sequences and protein secondary structure of LJFatB, LJCFatB, LHFatB and LDFatB are different and their proteins had conserved FatB substrate binding sites and catalytic activity sites. Transcriptive level of LJFatB, LJCFatB, LHFatB and LDFatB in bud was not significantly different. Therefore, LJFatB, LJCFatB, LHFatB and LDFatB could have the same biological function as AtFatB.

OBJECTIVETo study the function of the chalcone synthese gene introns in Scutellaria baicalensis, and clarify preliminarily their role in abiotic stress.

METHODThe CHS introns with specific primers were cloned and bioinformatic method was applied to predict the cis-elements in the intron of CHS. The introns were subcloned into binary vector, pCAMBIA-1301 before being transferred to tobacco. Then the activity of GUS of the transgenic tobacco seeds was analyzed.

RESULTSeven cis-elements were found in the introns. Under the dark and high temperature GUS expression rose at the first (3 h), but then declined (9 h). ABA and MeJA regulated insignificantly the GUS activity in normal temperature; treatment of 10% PEG induced GUS expression.

CONCLUSIONCHS introns could be play a role in the regulation of S. baicalensis phenylpropanoid biosynthetic pathway.

Acyltransferases ; genetics ; Base Sequence ; Gene Expression Regulation, Plant ; drug effects ; Gene Order ; Genetic Vectors ; Introns ; Molecular Sequence Data ; Plants, Genetically Modified ; genetics ; metabolism ; Polyethylene Glycols ; pharmacology ; Regulatory Sequences, Nucleic Acid ; genetics ; Scutellaria baicalensis ; enzymology ; genetics ; Sequence Alignment ; Tobacco ; genetics ; metabolism

Objective To evaluate virtual navigation assisted contrast-enhanced ultrasound(VN-CEUS) by comparing VN-CEUS with routine CEUS in detecting focal liver lesion(FLL) invisible in B-mode ultrasound(BUS).Methods Forty-seven FLLs invisible in BUS were identified by CT/MR,and received VN-CEUS and routine CEUS respectively.Results The detection rate of VN-CEUS and routine CEUS were 78.7%(37/47) and 36.2%(17/47),the former was much higher than the later (P<0.01).In all the cases,fusion of CT/MR with ultrasound was achieved successfully and average time for image fusion was 5 min.compared with routine CEUS,VN-CEUS could more easily find FLLs shadowed by lung or ribs,FLLs in inhomogeneous liver background,and FLLs with brief or inapparent abnormal enhancement.ConclusionsVN-CEUS can greatly improve the detection rate of FLL that was invisible in BUS.