AIM: To analyze the effect of pigment epithelium derived factor (PEDF) on nitrogen monoxide (NO) and expression of cysteine-containing, aspartate-specific proteases-3 (caspase-3) in retinal tissues of model rats with optic nerve crush injury.METHODS: A total of 60 SD rats were randomly divided into the blank control group, model group and PEDF group, with 20 rats in each group.Except the blank control group, the optic nerve crush injury rat models were established in the other groups, and left eyeballs were taken as samples.After successfully modeling, the model group were treated with intravitreal injection of 5μL of balanced salt solution while PEDF group were treated with intravitreal injection of 5μL of PEDF (0.2μg/μL).Two weeks later, the retinal tissues were collected, and changes of shape were observed under microscope after HE staining.The changes of NO level were measured by colorimetry assay, the expression of caspase-3 mRNA and caspase-3 protein was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western-blot.RESULTS: HE staining showed that retinal tissues of the blank control group arranged neatly and clearly.Retinal ganglion cells (RGCs) arranged in a monolayer, and cells were oval, uniform in size and distribution, the cell nuclei were clear, closely arranged, with clear boundaries.The retinal tissues of the model group were sparse in shape, RGCs showed vacuolar changes, the overall number of cells was reduced, and cell nuclei of residual RGCs showed pyknosis and uneven staining.RGCs in PEDF group were with slightly edema and arranged closely, and the degree of injury was significantly milder than that in the model group.Levels of Caspase-3 mRNA and protein and NO levels in the three groups showed the model group > PEDF group > blank control group (all P < 0.05).CONCLUSION: The application of PEDF can down regulate the expression of Caspase-3 and NO in rates with optic nerve injury and reduce RGCs injury.

Objective To study the characteristics of event-related potentials P300 in patients with anxiety disorder(AD). Methods P300 tests were carried out in 30 patients with AD and 30 healthy adult controls. ResultsPatients with AD had significantly delayed P3 latency ([326?16] ms vs [339?19]ms, P

OBJECTIVETo observe the three-dimensional distribution of vessels, and to establish a new method for measurement of blood flow velocity in mice cerebral cortex using two-photon laser scanning microscopy and fluorescence probe labeling technique.

METHODSThe mouse was made cranial window surgery and injected Texas-Red through tail vein after anesthetized. The three-dimensional imaging of vessel was obtained through z-stack scanning, and blood flow velocity was quantified through line scanning.

RESULTSWe could detect vascular distribution for more than 500 µm depth using two-photon microscopy. The velocity of blood flow was (0.59 ± 0.12) mm/s in capillary.

CONCLUSIONThe method for observing the brain blood flow by two-photon microscopy was established, which could achieve quantification of single vascular blood flow velocity and provide experimental evidence for basic research and medical applications.

Animals ; Blood Flow Velocity ; Brain ; blood supply ; Capillaries ; Cerebrovascular Circulation ; Fluorescent Dyes ; Hemodynamics ; Mice ; Microscopy, Fluorescence

OBJECTIVETo determine whether the sonic hedgehog (Shh) signaling could regulate the expression of histone demethylases in the head and neck squamous cell carcinoma(SCC).

METHODSHuman recombinant SHH-N protein or over-expression of the mutant 2 smoothened (M2-SMO) was applied to activate the Shh signaling in tongue squamous cell carcinoma cell line-SCC-6 in this study. Cyclopamine was used to block the Shh signaling in SCC-6. The real-time reverse transcription (RT)-PCR was used to detect the expression of histone demethylases at the mRNA level.

RESULTSThe data showed that activation of the Shh signaling up-regulated the expression of histone demethylase, lysine-specific demethylase 8 (KDM-8) at the mRNA level by human recombinant SHH-N protein (1.841 ∼ 3.591 fold compare with untreated group; P < 0.01), over-expression of the M2-SMO also increased the expression of KDM-8 (1.358 ∼ 3.013 fold compared with empty vector group; P < 0.05), and after the Shh signaling was blocked by Cyclopamine, the expression of KDM-8 was down regulated (decreased 25.6% ∼ 66.6% compared with control cells, P < 0.05).

CONCLUSIONSHistone demethylase KDM-8 was downstream target gene of Shh signaling in head and neck squamous cell carcinoma cell line SCC-6, and its expression was positively regulated by the Shh signaling.

Carcinoma, Squamous Cell ; genetics ; metabolism ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Head and Neck Neoplasms ; genetics ; metabolism ; Hedgehog Proteins ; metabolism ; Histone Demethylases ; genetics ; metabolism ; Humans ; Mutant Proteins ; metabolism ; RNA, Messenger ; genetics ; Receptors, G-Protein-Coupled ; metabolism ; Recombinant Proteins ; metabolism ; Signal Transduction ; Smoothened Receptor ; Veratrum Alkaloids ; pharmacology

Antibiotics, Antineoplastic ; pharmacology ; Breast Neoplasms ; enzymology ; pathology ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Female ; Glucosyltransferases ; biosynthesis ; genetics ; Humans ; Oligodeoxyribonucleotides, Antisense ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Transfection

The effect of human cytomegalovirus (HCMV) on invasive capability of early pregnant extravillous cytotrophoblasts (EVTs) was investigated in vitro. Primary EVTs were obtained by complex phosphoesterasum digestion and gradient centrifugation from villous tissue aseptically taken from healthy pregnant women. Cytokeratin7 (CK7), vimentin (Vim) and c-erbB-2 were immunocytochemically detected to identify source of cells, and HCMVpp65 antigen was assayed to determine the infection state of primary EVTs by immunocytochemical staining. The EVTs were divided into two groups: control group and HCMV group, and the expression of c-erbB-2, matrix metalloproteinase-2 (MMP-2) and MMP-9 proteins was detected in two groups by immunocytochemistry and Western blotting. Enzymic activity changes of MMP-2 and MMP-9 were tested by gelatin zymography in primary EVTs infected with HCMV. The invasion of primary EVTs was detected by cell invasion assay in vitro after they were infected by HCMV. The cell source identification showed that the cells obtained were highly-pure primary EVTs, and primary EVTs could be infected by HCMV. Primary EVTs could express c-erbB-2, MMP-2 and MMP-9 proteins, and as compared with control group, the protein expression was decreased significantly in HCMV groups (P<0.05). Primary EVTs could secrete active MMP-2 and MMP-9 in vitro, and the activity of two MMPs was decreased significantly in HCMV groups (P<0.05). The in vitro cell invasion assay showed that the number of primary EVTs permeating Matrigel in HCMV group was decreased (P<0.05). We are led to conclude that HCMV can infect primary EVTs and inhibit their invasion capability, suggesting that the impaired EVT's invasion capability might be related to the abnormal expression of c-erbB-2, MMP-2 and MMP-9 proteins.

An efficient modified route based on the targeting mechanism of antibacterial fluoroquinolones for the shift from the antibacterial activity to the antitumor one was further developed. Using a fused heterocyclic ring, s-triazolothiadiazine as a carboxyl bioisostere of ciprofloxacin, the title compounds, 1-cyclopropyl-6-fluoro-7-piperazin-1-yl-3-(6-substituted-phenyl-7H-[1, 2, 4]triazolo[3, 4-b][1, 3, 4]thiadiazin-3-yl)-quinolin-4(1H)-ones (5a-5e) and their corresponding N-acetyl products (6a-6e), were designed and synthesized, separately. Meaningfully, a ring-contraction of fused six-membered thiadiazine occurred by a sulfur extrusion reaction gave new tri-acetylated fused heterocycles related to pyrazolo[5, 1-c][1, 2, 4] triazoles (7a-7e). The in vitro antitumor activity against L1210, CHO and HL60 cell lines was also evaluated for the synthesized fifteen heterocycles compared to parent ciprofloxacin by methylthiazole trazolium (MTT) assay. Interestingly, the results displayed that fifteen fused heterocyclic compounds showed more significant growth inhibitory activity (IC50 < 25.0 micromo x L(-1)) than that of parent ciprofloxacin (IC50 > 150.0 micromol x L(-1)), and the active order decreased from 7a-7e to 5a-5e to 6a-6e, respective.
Animals ; Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; CHO Cells ; Cell Line, Tumor ; Ciprofloxacin ; pharmacology ; Cricetinae ; Cricetulus ; Fluoroquinolones ; chemical synthesis ; chemistry ; pharmacology ; HL-60 Cells ; Humans ; Inhibitory Concentration 50 ; Leukemia L1210 ; pathology ; Mice ; Structure-Activity Relationship ; Thiadiazines ; chemical synthesis ; chemistry ; pharmacology ; Triazoles ; chemical synthesis ; chemistry ; pharmacology

OBJECTIVETo investigate the prognostic predictors of nasal NK/T cell lymphoma.

METHODSThe clinicopathologic feature data of 61 patients with nasal NK/T cell lymphoma proven by pathological examination from Jan. 1997 to Jan. 2005 were collected. Expression of survivin, CD44, nm23, p53, Ki-67, MDR-1 and CD95 was detected by immunohistochemical staining in 30 patients with available histologic specimens. The correlation between these factors and prognosis were analyzed.

RESULTSIn univariate analysis, performance status, LDH level, clinical stage, initial treatment response, CD56, Ki-67 and CD95 were found to be the prognostic factors associated with time to progression (TTP) in nasal NK/T cell lymphoma, while the performance status, B symptoms, LDH level, initial treatment response, Ki-67 and CD95 were demonstrated as prognostic factors related to overall survival. In multivariate analysis, clinical stage, initial treatment response and performance status were independent prognostic factors for TTP, while the latter two factors were independent prognostic factors of overall survival.

CONCLUSIONClinical stage and initial treatment response, and performance status are found to be independent prognostic factors for TTP, whereas the latter two factors are demonstrated as independent prognostic factors of the overall survival. Overexpression of Ki-67 may be an unfavorable prognostic factor, but overexpression of CD95 may be a favorable one.

Adolescent ; Adult ; Aged ; Analysis of Variance ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Biomarkers, Tumor ; analysis ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Hyaluronan Receptors ; analysis ; Immunohistochemistry ; statistics & numerical data ; Inhibitor of Apoptosis Proteins ; Ki-67 Antigen ; analysis ; Killer Cells, Natural ; drug effects ; metabolism ; pathology ; Lymphoma, T-Cell ; drug therapy ; metabolism ; pathology ; Male ; Microtubule-Associated Proteins ; analysis ; Middle Aged ; Neoplasm Proteins ; analysis ; Neoplasm Staging ; Nose Neoplasms ; drug therapy ; metabolism ; pathology ; Prednisone ; therapeutic use ; Prognosis ; Proportional Hazards Models ; Vincristine ; therapeutic use ; fas Receptor ; analysis

OBJECTIVETo test expression level and glycosylation level of OPN in HCC cell lines with different metastatic potential and HCC tissues, and investigate the correlation between the glycosylation change and the liver cancer transporting as well as its significance.

METHODSThe level of OPN expression in liver cancer tissue(6 cases of non-metastasis and 7 cases of metastasis)as well as HCC cell lines with different metastatic potential (L02, Hep3B, MHCC97L, MHCC97H, HCCLM3, HCCLM6)was identified by immunohistochemistry and Western Blot, and then OPN was purified from HCC tissues by immunoprecipitation, followed by glycosylation detection of OPN from non-metastatic and metastatic HCC tissues by multiple lectin blot. Data were analyzed by t-test and variance analysis.

RESULTSDifferent levels of OPN expression were observed in HCC cell lines with different metastatic potential (F = 5.04, P = 0.008). Additionally, OPN expression level in HCC tissues with metastasis was higher than that in non-metastasis group (t = 2.447, P < 0.05). Relative optical density value was 0.69 ± 0.21 and 0.45 ± 0.14 respectively. OPN in liver cancer tissue was successfully purified using immunoprecipitation. Followed lectin blotting result showed that OPN protein in metastasis group showed lower affinity to MAL, PHAE, DSA, ConA as compared with that in non-metastasis group (P < 0.05).

CONCLUSIONSThe expression of OPN was positively correlated with the enhanced metastasis potential of HCC. OPN from metastasis HCC tissues presented lower level of some specific glycan structures such as a2, 3- sialic acid, bisecting GlcNAc, biantennary, muti-antennary and high mannose type N-glycan structure. This study not only indicates the role of OPN in HCC metastasis for the first time, but also provide experimental support for the mechanism of the function of OPN in the transportation of liver cancer cells as well as offer potential target for clinical treatment.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Glycosylation ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Neoplasm Metastasis ; Osteopontin ; metabolism

OBJECTIVETo develop an early and accurate detection method for hepatocellular carcinoma (HCC) based on detection of tumor-associated serum markers using a multiplex quantitative antibody array.

METHODSThe double-antibody sandwich principle was used to establish an antibody array composed of eight cancer-related serum markers, including alpha-fetoprotein (AFP), hepatocyte growth factor (HGF), insulin-like growth factor (IGF), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), transforming growth factor-beta 1 (TGF-b1), and vascular endothelial growth factor (VEGF). Serum samples from 160 cases of clinically diagnosed HCC and from 58 cases of liver cirrhosis (LC; controls) were obtained to test the array. Sixty percent of the samples were randomly selected for use as the training set (HCC, n = 96; LC, n = 36), and the remaining 40% was used as the test set (HCC, n = 64; LC, n = 22). The SPSS statistical software was used to perform logistic regression analysis and to create a diagnostic model.

RESULTSWhen used with the training set, the model had sensitivity of 93.3%, specificity of 83.3%, and accuracy of 90.9%. When used with the test set, the model had sensitivity of 89.0%, specificity of 77.3%, and accuracy of 86.0%. The traditional serum AFP value (cut-off value of 20 ng/mL) showed 70.0% diagnostic sensitivity, 59.0% specificity, and 64.0% accuracy.

CONCLUSIONThe newly developed multiplex quantitative antibody detection system has high sensitivity and specificity. The diagnostic model with AFP and seven other cancer-related factors was superior to the traditional AFP only approach for early diagnosis of liver cancer, indicating its potential clinical value.

Adult ; Aged ; Biomarkers, Tumor ; blood ; Carcinoma, Hepatocellular ; diagnosis ; Early Diagnosis ; Female ; Humans ; Liver Neoplasms ; diagnosis ; Male ; Microchip Analytical Procedures ; Middle Aged ; Sensitivity and Specificity ; Young Adult ; alpha-Fetoproteins ; immunology