Identification of differentially expressed genes in anagen dermal papilla by suppression subtractive hybridization / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>We constructed a cDNA subtractive library of dermal papilla cells (DPCs) in anagen with suppression subtractive hybridization (SSH) technique and clone differentially expressed genes related to DPCs in anagen.</p><p><b>METHODS</b>Total mRNA was isolated from DPCs of anagen and telogen follicles. Moreover, single-strand (ss) and double-strand (ds) cDNAs were synthesized in turn using SMART PCR cDNA synthesis technology. ds cDNAs then were digested with Rsa I and divided into two groups, and ligated to the specific adaptor 1 and adaptor 2R, respectively. After cDNAs were hybridized with each other twice and underwent two rounds of nested PCR. PCR products were ligated with arms of T/A plasmid vectors to set up the subtractive library. Selected clones were demonstrated by reverse Northern blot and sequenced. The acquired sequence data were aligned against the Genbank nucleotide database.</p><p><b>RESULTS</b>cDNA subtractive library of DPCs in anagen follicles was set up successfully with high subtractive efficiency. Thirty-five genes were identified in this study with 22 known functional genes and 13 unknown functional genes.</p><p><b>CONCLUSIONS</b>All results confirm the effectiveness and sensitivity of SSH in detecting differentially expressed genes from a small amount of clinical samples. Information about such alterations in gene expression could be useful for elucidating the genetic events in hair follicle growth regulation.</p>

Effect of anti-human VEGF monoclonal antibody on VEGF level in serum of nude mice transplanted buccal carcinoma: a quantitative study / 华西口腔医学杂志

<p><b>OBJECTIVE</b>Quantitative study of the effect of anti-human VEGF mAb E11 to VEGF level in serum of nude mice transplanted buccal carcinoma.</p><p><b>METHODS</b>E11 was administered into BALB/c nu/nu mice which were transplanted human buccal carcinoma. The saline was administrated as negative control. Mice were killed at 18 days. The VEGF level in serum of mice was determined by improved indirect ELISA.</p><p><b>RESULTS</b>Compared with the VEGF level in serum of mice in saline group, it was dramatically decreased in E11 group. The VEGF level in serum of mice treated E11 by subcutaneous was lowest and only reached (1.17 +/- 0.13) microg/L.</p><p><b>CONCLUSION</b>It demonstrated that the anti-human VEGF mAb could reduce the VEGF level in serum by binding VEGF, and block its biological activity. It indicates that VEGF in serum of malignant tumor patient is a new tumor marker.</p>

Study on serum-free culture of dermal papillae cells of human hair / 中华皮肤科杂志

Objective To explore the feasibility of culturing dermal papillae cells (DPC) of hu- man hair in a serum-flee medium,and to observe the growth characteristics of these cells.Methods Cell culture flasks (plates) were pretreated with fibronectin,and DPC (2nd passage) were incubated with Williams E serum-flee medium supplemented with insulin-transferrin-selenite (ITS).Cells were observed by an inverted phase-contrast microscope.Proliferation of DPC was evaluated with 3-(4,5-dimethylthia- zol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and by their growth curve.Results In a serum-free medium,2nd passage DPC adhered to the flask surface within two to four hours of incubation; two to three days later,confluence,of the cells was observed,without noticeable proliferation.Four days later,cell connection was interrupted,isolated cells or cell clusters were seen,and detachment of some cells from the flask surface was observed.One to two weeks later,most cells had died.After incubation with 4% bovine serum for ten hours,cell proliferation was observed surrounding the remaining viable cell colonies. DPC growth curve showed stagnant phase and slow growth phase;however,log growth phase was not ob- served.Conclusion DPC could be successfully cultured in serum-free medium.However,the culture con- dition needs to be further optimized.