The identification of differential gene expression between gray and black hairs is an important study in modern hair pigment research. In this experiment, the authors have applied new methods by the integration of three updated molecular biological tools, T7 RNA polymerase-based RNA amplification, representational difference analysis (RDA), and microarray analysis, to screen the differentially expressed genes in gray and black hairs. The genes more abundantly expressed in black hairs were pigment related proteins, such as Pmel17, 95kD melanocyte-specific secreted glycoprotein, MART-1, tyrosinase, tyrosinase-related protein 1 etc. Also, expression of the selenium-binding protein (hSBP) gene and the spast gene for spastin protein were up-regulated in black hairs compared to those in gray hairs. In gray hairs, many kinds of genes related with keratin, trichohyalin and transmembrane glycoprotein were more expressed. In particular ch 17, hRPK.142_H_19 was expressed in gray hairs as high signal intensity.
DNA* ; Gene Expression* ; Glycoproteins ; Hair* ; Mass Screening* ; Microarray Analysis ; Monophenol Monooxygenase ; Oligonucleotide Array Sequence Analysis* ; RNA

OBJECTIVETo study the expression of CD44v6 and E-cadherin(ED) in prostate carcinoma in relation to the metastasis of prostate carcinoma.

METHODSThe expression of CD44v6 and ED in 45 cases of prostate carcinoma was studied with immunohistochemical technique.

RESULTSThe positive expression rates of CD44v6 and ED in prostate carcinoma were 77.8% and 48.9%, respectively. The high level expression of CD44v6 and the low level expression of ED were positively correlated with differentiation, clinical staging and metastasis of prostate carcinoma (P < 0.05). The expression of CD44v6 was negatively correlated with ED(r = -0.58, P < 0.005).

CONCLUSIONSThe expression of CD44v6 and ED protein might be a useful marker for prostate carcinoma in evaluating the biological behavior and prognosis of the tumor.

Cadherins ; analysis ; Cell Differentiation ; Glycoproteins ; analysis ; Humans ; Hyaluronan Receptors ; analysis ; Immunohistochemistry ; Male ; Neoplasm Metastasis ; Prostatic Neoplasms ; chemistry ; pathology

Pterygium is a proliferative disease. Recent research has reported that stem cells are involved in the pathogenesis of various proliferative diseases, including solid tumors and diabetic proliferate vitreoretinopathy. In previous literature, we hypothesized that adult stem cells originated from bone marrow were involved in the pathogenesis of pterygium. We proved this by immunohistochemical staining with various stem cell markers. The staining showed adult stem cells in the pterygium. c-kit positive cells were observed primarily in the stroma, and some cells were also found in the basal epithelium. AC133 and CD34 positive cells were primarily found in the basal epithelium and were ovoid shaped, similar to the c-kit cells. However, some cells were found in vascular endothelium. STRO-1 positive cells were found mainly in the stroma and were spindle shaped. In recurrent pterygium, cells were more scattered and the expression pattern was denser. Therefore, we suggest a new theory of pterygium pathogenesis. Inflammation caused by environmental factors triggers the abnormal production of some growth factors and cytokines in order to recover from cellular damage. If these healing signals are excessive, limbal basal cells will be changed to abnormally-altered pterygial cells. The excessive wound healing process and remnant altered cells result in recurrence using the same mechanism.
Stem Cells/*physiology ; Pterygium/*etiology ; Proto-Oncogene Proteins c-kit/analysis ; Peptides/analysis ; Middle Aged ; Humans ; Glycoproteins/analysis ; Bone Marrow Cells/*physiology ; Antigens, CD34/analysis ; Antigens, CD/analysis

We purified a novel mannose binding lectin form Musca domestica pupae by affinity chromatography on Con A-Sepharose 4B and DEAE weak anion-exchange chromatography. By SDS-PAGE, MBL-1 yielded a single band with the molecular weight of 24 kDa. It was a glycoprotein detected by periodic acid-schiffs staining reaction, with 97.36% protein and 2.1% oligosaccharide. Meanwhile, the results of beta-elimination reaction, infrared spectroscopy, atomic force microscopy and protein sequencing instrument show that MBL-1 was an ellipsoidal-shaped monomer with 60-100 nm in diameter. N-glycoside bond linked oligosaccharide chain and the N-terminal blocked peptide chain. Further study suggested that MBL-1 promote the proliferation of macrophage in a concentration-dependent manner. The scanning electron microscope analysis shows that MBL-1 promoted the activation of macrophages. These results show that MBL-1 purified from Musca domestica pupae possesses immune regulation effect, serving a reference basis to develop natural immune-modulator.
Animals ; Glycoproteins ; analysis ; Houseflies ; chemistry ; Immunomodulation ; immunology ; physiology ; Macrophages ; immunology ; Mannose-Binding Lectin ; chemistry ; physiology ; Oligosaccharides ; analysis ; Pupa ; chemistry

BACKGROUND/AIMS: Cluster differentiation 44 standard isoform (CD44s) is a transmembrane glycoprotein. CD44s is a known prognostic factor in various cancers, due to its involvement in tumor cell growth, invasion and metastasis. Its prognostic role, however, is debated because it can be a positive or negative prognostic factor depending on tumor type and is still an ambiguous prognostic indicator in other cancers, especially hepatocellular carcinoma (HCC). We investigated the relationship between CD44s expression and survival in HCC patients. METHODS: A total of 260 HCC samples were collected to generate a tissue microarray. Staining of the arrays with a primary mouse CD44s monoclonal antibody was followed by evaluation of the relationship between CD44s expression and tumor differentiation. The effect of CD44s expression on patient survival was analyzed. RESULTS: CD44s protein expression correlated with histological grade (most and worst Edmondson grade) of the HCC (p=0.029 and p=0.039, respectively) and adversely affected the disease free survival period based on univariate and multivariate analyses (p=0.038 and p=0.077, respectively). CONCLUSIONS: High CD44s protein expression correlates with shorter disease free survival and poorly differentiated HCC. CD44s-targeted therapy may be efficacious for HCC treatment in the future.
Animals ; Antigens, CD44 ; Carcinoma, Hepatocellular ; Disease-Free Survival ; Glycoproteins ; Humans ; Mice ; Multivariate Analysis ; Neoplasm Metastasis ; Protein Array Analysis ; Recurrence

OBJECTIVETo investigate the expression of cluster of differentiation 44 variant 6 (CD(44V6)) and proliferating cell nuclear antigen (PCNA) in ocular squamous cell carcinomas.

METHODSStreptavidin-biotin complex (SABC) immunohistochemistry was used to explore the expression of CD(44V6) and PCNA in 35 cases of ocular squamous cell carcinomas, 20 cases of papillomas, and 11 cases of normal eyelid tissue.

RESULTSThe CD(44V6) positive rate was 62.9% (22/35) in ocular squamous cell carcinomas, 15.0% (3/20) in papillomas, but not detectable in the 11 cases of normal eyelid tissue. The positive expression rates of CD(44V6) in ocular squamous cell carcinomas were significantly higher than in benign tumors (chi(2) = 11.57, P < 0.01) or control tissue (P = 0.001), and the positive expression rates of CD(44V6) in metastasis were significantly higher than without metastasis (P = 0.049). PCNA labeling indexes (PI) in tumors with CD(44V6) expression were significantly higher than those without (t = 20.21, P < 0.01).

CONCLUSIONSOverexpression of CD(44V6) is correlated with the progress and metastasis of ocular squamous cell carcinomas. CD(44V6) protein positive staining is associated with high PI. CD(44V6) and PCNA are useful for evaluating prognosis.

Carcinoma, Squamous Cell ; chemistry ; pathology ; Eye Neoplasms ; chemistry ; pathology ; Glycoproteins ; analysis ; Humans ; Hyaluronan Receptors ; analysis ; Immunohistochemistry ; Lymphatic Metastasis ; Proliferating Cell Nuclear Antigen ; analysis ; Skin ; chemistry

This study was to investigate dynamics of biological properties of CD133(+) cells from human umbilical cord blood (UCB) during short-term culture containing the combination of hematopoietic growth factors and the feasibility of in vitro expansion of CD133(+) cells. The biology activities including analysis of cell cycle, immunophenotype, telomerase activity, expression of adhesion molecules and expansion potential of CD133(+) cells were monitored during ex-vivo expansion, and compared with those of CD34(+) cells. The results showed that the contents of CD133(+) and CD34(+) cells in fresh UCB were (1.05 +/- 0.73)% and (1.40 +/- 0.56)% respectively. About 79.62% of CD34(+) cells expressed CD133, and more than 97% of CD133(+) cells were CD133(+)CD34(+), markedly higher than that in CD34(+) fraction (P < 0.01). No significant differences were observed in content of cells expressing CD38, CD13, CD14, CD61 and glycophorin-A between the two fractions. Expansion of CD133(+), CD133(+)CD34(+) and CD34(+)CD38(-) cells at 10 days and those of CFU-mix, HPP-CFC and CD34(+)CD38(-) cells at 6 days from CD133(+) cells group were significantly higher than those from the CD34(+) cell group (P < 0.05). Analysis of immunophenotype showed that CD133(+)CD34(+) cells declined gradually while CD133(-)CD34(+) and CD133(-)CD34(-) cells increased during ex-vivo expansion; basal telomerase activities of fresh UCB CD133(+) and CD34(+) cells were low but significantly exceeded that of CD34(-) fraction (P < 0.05). At first week of expansion, telomerase activity was significantly upregulated, after two weeks, telomerase activity remarkably declined, and decreased to baseline or below the limits of detection in day 20. More than 90% of CD133(+) cells expressed CD49d and CD11a, and, more than 85% of the cells expressed CD54, about 50% of cells expressed CD62L. At the early stage of expansion, expression of CD49d was upregulated, expression of CD11a remaining no change, while as expression of CD54 and CD62L was downregulated. Expression of all adhesion molecules was decreased gradually with extend of culture. But expression of these adhesion molecules on CD34(+) subsets were not affected significantly during expansion. It is concluded that CD133(+) population may be a more primitive hematopoietic stem/progenitor cells (HSPC) than CD34(+) cells, CD133(+) cells have great expansion potential for ex-vivo expansion and is a suitable target cell for ex-vivo expansion of HSPC. Downregulation of adhesion molecules and telomerase activity may be one of the reasons for delayed engraftment of expanded products.
AC133 Antigen ; Antigens, CD ; Antigens, CD34 ; analysis ; Cell Cycle ; Cells, Cultured ; Fetal Blood ; cytology ; Glycoproteins ; analysis ; Hematopoietic Stem Cells ; physiology ; Humans ; Immunophenotyping ; Peptides ; analysis ; Telomerase ; metabolism

This study aimed to investigate the clinical effect of transplantation of CD133⁺ peripheral blood stem cells or umbilical cord mesenchymal stem cells via the hepatic artery in children with type II hyperammonemia and its possible action mechanism. Umbilical cord mesenchymal stem cells were obtained by collecting cord blood (100-150 mL) from healthy fetuses and separating stem cell suspension (5 mL) from the cord blood by hydroxyethyl starch sedimentation. CD133⁺ peripheral blood stem cells were obtained by mobilizing peripheral blood from the fathers of sick children using recombinant human granulocyte colony-stimulating factor for 5 days, collecting mononuclear cells (120 mL), and separating out CD133⁺ cells by sorting. With catheterization and percutaneous puncture, the obtained stem cells were slowly injected into the liver of sick children via the hepatic artery. The changes in clinical symptoms and laboratory indices such as blood ammonia, liver function, and arginine and citrulline concentrations were observed. After stem cell transplantation via the hepatic artery, the 6 children showed significantly decreased blood ammonia levels, and their blood ammonia levels slowly increased 1 to 2 weeks later, but remained below 100 μmol/L, and changes in glutamic-pyruvic transaminase levels were similar to blood ammonia. Plasma citrulline and arginine concentrations increased significantly after transplantation and the increase in citrulline level exceeded the increase in arginine level. An 8 months follow-up visit for one typical patient showed that the weight and height increased after transplantation and sleep was improved without night crying. The child could actively gaze at interesting objects instead of responding indifferently and started to say simple words. With regard to fine motor skills, the child could pinch things with the thumb and middle finger instead of displaying a lack of hand-eye coordination and progress was also made in gross motor skills. Gesell test showed that the child made progress for an average of 3.82 months in all areas. It was concluded that after stem cell transplantation, children with type II hyperammonemia have decreased blood ammonia levels, stable and improved liver function and steadily increased plasma citrulline and arginine concentrations. They display a progressive trend in such aspects as movement, language and environmental adaptability. It is hypothesized that stem cell transplantation via the hepatic artery partially or totally activates, or provides supplementary ornithine carbamoyl transferase, so that plasma citrulline and arginine concentrations increase and urea cycle disorder can be corrected to some extent.
AC133 Antigen ; Ammonia ; blood ; Antigens, CD ; analysis ; Arginine ; blood ; Citrulline ; blood ; Female ; Glycoproteins ; analysis ; Hepatic Artery ; Humans ; Hyperammonemia ; blood ; surgery ; Infant ; Male ; Peptides ; analysis ; Stem Cell Transplantation

OBJECTIVETo investigate the host-defense role of short palate, lung, and nasal epithelium clone 1 (SPLUNC1) in Streptococcus pneumoniae (SP) infection and the effect of resveratrol (Res) on SPLUNC1 expression, and to provide new thoughts for the treatment of diseases caused by SP infection.

METHODSAccording to the multiplicity of infection (MOI), BEAS-2B cells with SP infection were divided into control group, MOI20 SP group, and MOI50 SP group. According to the different concentrations of Res, the BEAS-2B cells with MOI20 SP infection pretreated by Res were divided into 12.5Res+SP group, 25Res+SP group, and 50Res+SP group (the final concentrations of Res were 12.5, 25, and 50 μmol/L, respectively). Cell Counting Kit-8 was used to measure cell activity and determine the optimal concentration and action time of SP and Res. In the formal experiment, the cells were divided into control group, Res group, SP group, and Res+SP group. Real-time PCR and ELISA were used to measure the mRNA and protein expression of SPLUNC1.

RESULTSOver the time of SP infection, cell activity tended to decrease. Compared with the control group and the MOI20 SP group, the MOI50 SP group had a reduction in cell activity. Compared with the MOI20 SP group, the 25Res+SP group had increased cell activity and the 50Res+SP group had reduced cell activity (P<0.05). MOI20 SP bacterial suspension and 25 μmol/L Res were used for the formal experiment. Over the time of SP infection, the mRNA expression of SPLUNC1 in BEAS-2B cells firstly increased and then decreased in the SP group and the Res+SP group (P<0.05). Compared with the SP group, the Res+SP group had significant increases in the mRNA and protein expression of SPLUNC1 at all time points (P<0.05). Compared with the control group, the Res group had no significant changes in the mRNA and protein expression of SPLUNC1 (P>0.05).

CONCLUSIONSSP infection can induce SPLUNC1 expression and the host-defense role of SPLUNC1. Res can upregulate SPLUNC1 expression during the development of infection and enhance cell protection in a concentration- and time-dependent manner.

Bronchi ; metabolism ; Cells, Cultured ; Cytoprotection ; Epithelial Cells ; metabolism ; Glycoproteins ; analysis ; genetics ; physiology ; Humans ; Phosphoproteins ; analysis ; genetics ; physiology ; RNA, Messenger ; analysis ; Stilbenes ; pharmacology ; Streptococcus pneumoniae ; pathogenicity

OBJECTIVETo determine AF172993 sequence is either the complete CDS or a transcript variant.

METHODSRT-PCR was used to amplify the CDS sequence of Plunc, which was subsequently cloned into the pEGFP-N1 eukaryotic expression vector. After bi-directional sequence analysis, the sequence obtained was blasted against the AF172993 sequence, nr database and human genome database.

RESULTSIn CDS of the new cloned sequence, the 658 base A in the AF172993 sequence was replaced by C, and the corresponding genetic code was also converted from AAG to CAG, leading to the alteration of the amino acid Gln to Lys. In addition, the base C at the 658 position of the CDS showed perfect match with the base C at 2094188 position in human chromosome 20.

CONCLUSIONThe base A at the 658 position of AF172993 sequence of Plunc is a mutation site, which alters the coding of the amino acid. AF172993 sequence is actually a transcript variant of Plunc, and the annotation to AF172993 in GenBank database is not correct and need to be revised.

Cloning, Molecular ; Databases, Nucleic Acid ; Glycoproteins ; genetics ; Humans ; Mutation, Missense ; Open Reading Frames ; genetics ; Phosphoproteins ; genetics ; Sequence Analysis, DNA