INTRODUCTIONWe studied the automated intraretinal segmentation on Fourier domain optical coherence tomography (OCT).

MATERIALS AND METHODSThirty eyes from 30 normal subjects were studied using the RTVue-100. Both radial and raster scan protocol were performed 3 times on each subject. The OCT software performs automated intraretinal segmentation and provides macular thickness measurements.

RESULTSBoth scanning protocols provide reproducible inner, outer and full retinal thickness measurements. The inner, outer and full retinal thicknesses at the foveal central subfield were 67.31 +/- 12.27 microm, 151.67 +/- 12.96 microm, 219.33 +/- 23.19 microm, respectively by the raster scan, and 63.27 +/- 10.37 microm, 147.07 +/- 14.54 microm, 209.89 +/- 21.80 microm, respectively by the radial scan. Macular regional variations were consistently observed. The raster scan protocol gives greater retinal thickness measurements than the radial scan protocol (P <0.05), but the latter yields slightly more reproducible results.

CONCLUSIONSFourier domain OCT equipped with the ability to perform automatic intraretinal segmentation is a convenient tool in studying diseases that may differentially affect various parts of the retina. However, the establishment of normative values can be complicated by different scanning protocols, devices used, methods of data presentation and definition of intraretinal boundaries.

Adult ; Humans ; Image Interpretation, Computer-Assisted ; methods ; Macula Lutea ; anatomy & histology ; Middle Aged ; Reference Values ; Retinal Diseases ; diagnosis ; Tomography, Optical Coherence ; instrumentation ; methods ; Young Adult

AIM(1) To investigate the mRNA expression of the key angiogenic growth factors in the grafts after transplantation. (2) To investigate the potential impact of danshen (Chinese traditional medicine) administration on grafts angiogenesis.

METHODSThe frozen-thawed ovarian tissue from aborted fetus were xenografted into the renal capsule of the nude mice, recovered 48 h, 7 d and 28 d after respectively. Either danshen or saline (as the control) was administered after transplantation.

RESULTSThe mRNA levels of VEGF showed a temporary raise in 48 h after transplantation, then decreased in one week, and no significant difference was fund between the control group and danshen group. Ang-2 was increased in 48 h after transplantation, when Danshen group was significantly higher than the control group (P < 0.05). The microvessel density significantly increased in all the tissues after transplantation. The control group peaked on day 7 after transplantation, while danshen group peaked in 48 h and kept correspondingly steady after that.

CONCLUSIONEarly angiogenesis began within 48 h after transplantation of the thawed human fetal ovarian tissue, and its microvessel density peaked within the first week after transplantation. Our results also suggested that the use of danshen injection in conjunction with transplantation could facilitate revascularization of the grafts.

Angiopoietin-2 ; genetics ; metabolism ; Animals ; Cryopreservation ; Drugs, Chinese Herbal ; pharmacology ; Female ; Fetal Tissue Transplantation ; methods ; Fetus ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neovascularization, Physiologic ; drug effects ; Ovarian Follicle ; cytology ; growth & development ; transplantation ; RNA, Messenger ; genetics ; metabolism ; Salvia miltiorrhiza ; chemistry ; Transplantation, Heterologous ; methods ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

Animals ; Blotting, Western ; Cell Differentiation ; drug effects ; Embryonic Stem Cells ; cytology ; enzymology ; Isoenzymes ; analysis ; Mice ; Neurons ; enzymology ; Protein Kinase C ; analysis ; Tretinoin ; pharmacology

This study was aimed to investigate the expression level of perforin in cord blood NK cells and the relation of perforin expression after IL-2, IL-15 stimulation to cytotoxicity of NK cells. NK cells were isolated from cord blood MNC by depleting CD3(+) cells and then enriching CD56(+) cells using immunomagnetic separation (CD3 and CD56 cell isolation kit, autoMACS, miltenyi). The purity was analysed by flow cytometry. According to the different combination of cytokines, there were two groups: group A (IL-2) and group B (IL-2 + IL-15). The cytotoxicity and perforin expression rate of fresh and different cultured CB-NK cells against K562/Jurkat cell lines were estimated by LDH release test (cytotoxic 96 non-radioactive cytotoxicity assay). The results showed that the purity of NK cells after separation was more than 90%. The cytotoxicity towards both tumor lines in group B was higher than that in group A (p < 0.05), and cytotoxicity in group A was higher than that of fresh NK cells (p < 0.05). Perforin expression rate of group A (84.55%) was higher than that of fresh NK cells (67.21%) (p < 0.05), and there was no significant difference between group A and B (84.55% versus 87.22%) Cytotoxic activity of CB-NK cells was positively correlated with perforin expression rate (r = 0.886, p < 0.05). It is concluded that IL-2 can enhance cytotoxicity of CB/BM-NK cells by increasing the perforin expression.
CD56 Antigen ; metabolism ; Cells, Cultured ; Cytotoxicity, Immunologic ; Fetal Blood ; cytology ; Flow Cytometry ; Humans ; Interleukin-15 ; pharmacology ; Interleukin-2 ; pharmacology ; K562 Cells ; Killer Cells, Natural ; cytology ; immunology ; metabolism ; Perforin ; metabolism

The aim of this study was to explore the cytotoxicity of fresh cord blood(CB) NK cells and the influence of IL-12 and IL-15 on activity of the NK cells killing K562 and Jurkat cells lines. The NK cells were isolated from cord blood by depleting CD3(+) cells and then enriching CD56(+) cells using sorting with immunomagnetic beads. The experiment was divided into 3 groups: group A (fresh CB-NK cells without cytokines), group B (CB-NK cells cultured by IL-2) and group C (CB-NK cells cultured by IL-2 and IL-15). The purity of NK cells was determined by flow cytometry; the cytotoxity of fresh and different cytokine-treated CB-NK cells on K562 and Jurkat cell lines was detected by LDH release test. The results showed that the purity of NK cells before and after sorting was 14.88 ± 9.2% and 92.39 ± 0.8% respectively. After culture for 3 days, NK-forming colony amounts in group B and group C were 148.60 ± 13.0 and 831.80 ± 23.0 respectively, the comparison between group B and group C showed the significant difference (p < 0.05). The cytotoxicities of NK cells in group A, B and C on K562 and Jurkat cell lines were 27.76 ± 8.8%, 61.90 ± 9.1% and 87.62 ± 3.7%; 29.32 ± 2.5%, 69.43 ± 4.4% and 92.95 ± 3.2% respectively, the difference was significant (p < 0.05). It is concluded that the fresh isolated CB-NK cells show low cytotoxic activity. After stimulated with IL-2 or IL-2 plus IL15, cytotoxicity of CB-NK cells increases obviously, the effect of IL-2 plus IL-15 is much better than IL-2 alone for promoting the growth and enhancing the cytotoxicity of CB-NK cells.
Cytotoxicity, Immunologic ; drug effects ; immunology ; Fetal Blood ; drug effects ; immunology ; Humans ; Interleukin-15 ; pharmacology ; Interleukin-2 ; pharmacology ; Jurkat Cells ; K562 Cells ; Killer Cells, Natural ; drug effects ; immunology

OBJECTIVEThe purpose of this study was to investigate the clinical significance of THY1 protein expression in epithelial ovarian cancer.

METHODSImmunohistochemistry (IHC) and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining were used to detect the protein expression of THY1, Ki67 and cell apoptosis in 76 epithelial ovarian cancers by tissue microarray. The correlation between THY1 expression and patients' clinical features was analyzed.

RESULTSOf the 76 epithelial ovarian cancer samples, 64 were informative for IHC and TUNEL assays and 42 (65.6%) among them showed down-regulated/loss expression of THY1 protein. A significant positive correlation of THY1 protein expression with clinical stage and distant metastasis was observed in this ovarian cancer cohort (P < 0.05). The more advanced the tumor stage, the more frequency of loss expression of THY1 protein. In addition, the mean positive rate of Ki67 staining in tumors with down-regulated/loss expression of THY1 was 33.7% +/- 3.5%, significantly higher than that in the tumors with normal expression of THY1 (17.3% +/- 6.1%, P = 0.0027). However, no significant correlation was observed between THY1 protein expression and tumor cell apoptosis as well as patients' survival in this series (P > 0.05).

CONCLUSIONDown-regulated/loss expression of THY1 protein in epithelial ovarian cancer is significantly correlated with cancer cell proliferation and metastasis in the epithelial ovarian cancer, and it may be used as one of the new molecular biomarkers to predict the disease progression in patients.

Adult ; Aged ; Apoptosis ; Cystadenocarcinoma, Mucinous ; metabolism ; pathology ; Cystadenocarcinoma, Serous ; metabolism ; pathology ; Down-Regulation ; Female ; Follow-Up Studies ; Gene Expression Regulation, Neoplastic ; Humans ; Ki-67 Antigen ; metabolism ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Staging ; Ovarian Neoplasms ; metabolism ; pathology ; Survival Rate ; Thy-1 Antigens ; metabolism

OBJECTIVETo investigate the expression of interleukin-8 (IL-8) and its prognostic significance in breast cancer.

METHODSExpression of IL-8 in 113 breast cancers, 19 breast benign tumors and 20 breast normal tissues was examined by tissue microarray using immunohistochemistry, and the association of IL-8 expression with patient's clinico-pathological characteristics and prognosis was further analyzed.

RESULTSThe positive rate of IL-8 expression in breast cancer was 27.4%, which was significantly higher than that in benign tumor and normal tissue of breast (P = 0.002). IL-8 expression related to histological type (P = 0.040) and lymph node status (P = 0.021). The expression of IL-8 was observed to correlate negatively with ER and PR status (P = 0.015 and P = 0.034), and correlate positively with C-erbB-2 status (P = 0.002). In addition, Kaplan-Meier curves of disease-free survival analysis showed a significant difference between IL-8 positive groups and negative group (P = 0.031).

CONCLUSIONSIL-8 might be a poor prognostic factor for human breast cancer, and also might be a novel molecular marker to predicate the occurrence and progression of breast cancer.

Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; diagnosis ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Interleukin-8 ; biosynthesis ; Lymphatic Metastasis ; Middle Aged ; Prognosis ; Receptor, ErbB-2 ; biosynthesis ; Receptors, Estrogen ; biosynthesis ; Receptors, Progesterone ; biosynthesis

MECP2 duplication syndrome (MDS) is a rare pediatric disease and mainly manifests as delayed motor development, language loss or delay, recurrent infection, severe intellectual disability, epilepsy, autistic symptoms, and early infantile hypotonia. In this article, the three children with this disease were all boys. Cases 1 and 2 had delayed motor development, and language loss or delay as initial manifestations, and case 3 had recurrent infection as initial manifestation. Physical examination showed hypotonia and negative pathological signs in each case. Case 1 had tonic-clonic seizures and electroencephalography showed focal seizures, for which he was given oxcarbazepine, levetiracetam, and clonazepam as the antiepileptic treatment to control seizures. Case 3 experienced one absence seizure and three head-nodding seizures with normal electroencephalographic findings during these seizures, and therefore, he was not given antiepileptic treatment. In each case, recurrent infection was improved with the increase in age, but there were no significant improvements in language or intelligence. Array-based comparative genomic hybridization (aCGH) showed MECP2 duplication in X chromosome in each case, and so they were diagnosed with MDS. MDS should be considered for children with delayed development complicated by recurrent infection and epileptic seizures, and early aCGH helps with the diagnosis of this disease.
Child ; Comparative Genomic Hybridization ; Humans ; Infant ; Male ; Mental Retardation, X-Linked ; complications ; genetics ; Methyl-CpG-Binding Protein 2 ; genetics

The pathological processes of Alzheimer's disease and type 2 diabetes mellitus have been demonstrated to be linked together. Both PDE9 inhibitors and PPAR agonists such as rosiglitazone exhibited remarkable preclinical and clinical treatment effects for these two diseases. In this study, a series of PDE9 inhibitors combining the pharmacophore of rosiglitazone were discovered. All the compounds possessed remarkable affinities towards PDE9 and four of them have the IC values <5 nmol/L. In addition, these four compounds showed low cell toxicity in human SH-SY5Y neuroblastoma cells. Compound , the most effective one, gave the IC of 1.1 nmol/L towards PDE9, which is significantly better than the reference compounds PF-04447943 and BAY 73-6691. The analysis of putative binding patterns and binding free energy of the designed compounds with PDE9 may explain the structure-activity relationships and provide evidence for further structural modifications.

To investigate the methylation status of CpG islands of the secreted frizzled-related protein (SFRP) gene promoter region in malignant hematopoietic cell lines, and to explore the possible relationship of CpG abnormal methylation status with pathogenic mechanism of hematologic malignancies. Methylation-specific PCR was used to detect the status of SFRP gene promoter region in nine malignant hematopoietic cell lines and peripheral blood mononuclear cells from healthy people. The results indicated that hypermethylation of 2 genes coding for SFRP1 and 2 were present in nine malignant hematopoietic cell lines, however, methylation and unmethylation of SFRP4 were both detected in CA46, HL60 and U937 cell lines, and SFRP5 in U266 as well. None of the normal mononuclear cells showed methylation of SFRP 1-5 genes. It is concluded that the hypermethylation of SFRP genes is related to the evolution of malignant hematopoiesis. Methylation of SFRP genes may serve as potential independent biomarkers for early detection of hematologic malignancies.
Cell Line, Tumor ; CpG Islands ; DNA Methylation ; Hematologic Neoplasms ; genetics ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; Membrane Proteins ; genetics ; Promoter Regions, Genetic ; Proto-Oncogene Proteins ; genetics