OBJECTIVETo investigate telomerase activity and expression of hTR and hTERT in human neuroepithelial tumors for exploring new strategy for clinical diagnosis and treatment.

METHODSTelomerase activity was detected by modified TRAP method and the expression of hTR and hTERT was measured by RT-PCR method in 65 human neuroepithelial tumors, respectively.

RESULTSThe positive rates of telomerase and hTERT were 61.54% and 70.77% respectively in human neuroepithelial tumors, and the positive rate and their level of expression were correlated with the degree of malignancy of tumors positively.

CONCLUSIONSTelomerase activity and hTERT are significantly correlated with the degree of malignancyin human neuroepithelial tumors. hTERT may play a key role in the regulation of telomerase activity.

DNA-Binding Proteins ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Neoplastic ; Humans ; Neoplasms, Neuroepithelial ; enzymology ; genetics ; Telomerase ; biosynthesis ; genetics ; metabolism

Angiotensin II ; immunology ; Chronic Disease ; Heart Failure ; drug therapy ; Humans ; Renin-Angiotensin System ; immunology ; Vaccines ; therapeutic use

OBJECTIVETo dynamically investigate the morphology of human gastric cancer SGC-7901 cell clones, and then compare the tumorigenic ability of different clones in order to identify the tumor stem cell clones.

METHODSClones derived from gastric cancer SGC-7901 cells were assessed by morphological observation, and the clone formation rate and proportion of each clone were calculated. The expression of CD44 and CDX2 in different clones was detected by immunofluorescence microscopy and Western blot. Furthermore, different clones were isolated and cultured, and their self-renewal property was assayed. Cells of different clones were subcutaneously inoculated into nude mice and the tumorigenic ability of each group was determined.

RESULTSClones derived from gastric cancer SGC-7901 cells had three types, i.e. clones of tight, transitional and loose types. The total clone formation rate was (9.80 ± 1.07)%, and the proportion of tight, transitional and loose type clones was 10.2%, 56.0% and 33.8%, respectively. The results of immunofluorescence microscopic examination showed that the signal of CD44 was significantly stronger in the tight clones than in the transitional and loose clones, however, the signal of CDX2 was weakest in the tight colonies. The results of Western blot were consistent with that of immunofluorescence microscopic observation. SGC-7901 cells of tight clones possessed strong ability of self-renewal and in vivo tumorigenicity in the nude mice.

CONCLUSIONSGC-7901 cell clones vary in morphology and differentiation, and the tight type clones may include rich gastric cancer stem cells.

Animals ; CDX2 Transcription Factor ; Cell Differentiation ; Cell Line, Tumor ; Cell Proliferation ; Clone Cells ; classification ; Female ; Homeodomain Proteins ; metabolism ; Humans ; Hyaluronan Receptors ; metabolism ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Neoplastic Stem Cells ; cytology ; metabolism ; Random Allocation ; Stomach Neoplasms ; metabolism ; pathology

OBJECTIVETo analyze the effects of high-frequency electromagnetic field (HF-EMF, 30 MHz, 0-1600 V/m) on the apoptosis and ultramicrostructure of the hippocamp and demonstrate the cytotoxicity of hippocamp.

METHODS120 Wistar female adult rats were randomly divided into ten groups based on body weight with different levels of 30 MHz electromagnetic field (0, 25, 100, 400, 1600 V/m) for eight hours daily. Five group rats were irradiated for three days. The other five group rats were irradiated for fifty-six days. Weekly the rats were continuously exposed five days. The apoptotic rate of the hippocamp was detected with TUNEL System. Meanwhile, the ultramicrostructure was observed with the transmission electron microscope.

RESULTS(1) There was no significant difference on the apoptotic rate and pathological change of the hippocamp cell between the exposure and the control groups through short term experiment (P > 0.05). (2) The apoptotic rate of the granulocyte on the DG campus of the hippocamp in the 400 V/m group and the 1600 V/m group (0.165% +/- 0.049%, 0.189% +/- 0.049% respectively) were increased significantly (P < 0.01) through inferior chronic experiment compared with the control group (0.052% +/- 0.016%). Along with the increase of radiation dose, the ultramicrostructure of the neuron cell appeared more abnormal cells. Especially there were marked change on the neuron in the 1600 V/m group.

CONCLUSIONSThere is no association between cell apoptotic rate of the hippocamp and short period exposure to HF-EMF (30 MHz, 25-1600 V/m). However inferior chronic exposures to HF-EMF might induce the cytotoxicity, especially in the high dose exposure (1600 V/m) under our experiment.

Animals ; Apoptosis ; radiation effects ; Electromagnetic Fields ; Endocytosis ; radiation effects ; Female ; Hippocampus ; cytology ; pathology ; radiation effects ; Neurons ; pathology ; radiation effects ; Rats ; Rats, Wistar

OBJECTIVETo select three kinds of perforation repair materials, mineral trioxide aggregate (MTA), Z350, amalgam. And to evaluate the cytotoxicity of three kinds of perforation repair materials on human periodontal ligament fibroblasts (HPDLF) in vitro.

METHODSThe proliferation of HPDLF to three perforation repair materials were examined by methyl thiazolyl tetrazolium (MTT) assay at 1, 3 and 5 days. The mRNA expression levels of bone-associated alkaline phosphatase (ALP) and osteocalcin (OC) were determined using a real-time quantitative polymerase chain reaction (PCR).

RESULTSMTA shew almost no inhibition to HPDLF, the expression of ALP mRNA and OC mRNA in the HPDLF cultured on MTA were higher. Z350 induced a slight inhibition to HPDLF, and the expression of ALP mRNA but there was no difference in the expression of OC mRNA. Cell proliferation was significantly impaired by amalgam with grade 3, and the expression of ALP mRNA and OC mRNA were significantly reduced.

CONCLUSIONMTA have minimum cytotoxicity on HPDLF and can promote cell differentiation and regenerate of periodontal tissue. Z350 have lower cytotoxicity on HPDLF. Amalgam show highest cytotoxicity on HPDLF in the three materials and inhibit cells differentiation.

Acrylic Resins ; Aluminum Compounds ; Calcium Compounds ; Cell Differentiation ; Cells, Cultured ; Drug Combinations ; Fibroblasts ; Humans ; In Vitro Techniques ; Osteocalcin ; Oxides ; Periodontal Ligament ; Root Canal Filling Materials ; Silicates

OBJECTIVESTo detect the expressions of Tec tyrosine kinase in hepatocellular carcinoma and the levels of phosphorylation of tyrosine kinase in liver cancer tissues, paracancerous tissues and normal liver tissues and to find the significance of their differences.

METHODS200 specimens of tissues, including liver cancer tissues, surrounding liver tissues not more than 1.5 cm from the cancers, and normal liver tissues were investigated for Tec protein expression and Tec phosphorylation by tissue microarrays and immunohistochemistry (SP method).

RESULTSThe positive immunohistochemical stainings of Tec in cancerous tissues and non-cancerous tissues showed no obvious differences, nevertheless, the immunostaining levels in liver cancer tissues were much higher than in non-cancerous tissues and they correlated with the grading of tumors (P < 0.05). The phosphorylation of Tec was significantly expressed in liver cancer tissues (73%) in comparison with other tissues (42%, 10% both P < 0.05), but it did not correlate with any clinicopathological characteristics.

CONCLUSIONOverexpression of Tec is associated with the tumorigenesis and development of liver cancer; inhibiting Tec or degrading Tec phosphorylation directly might affect the progression of liver cancer.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Hepatocellular ; metabolism ; Female ; Humans ; Liver Neoplasms ; metabolism ; Male ; Middle Aged ; Neoplasm Proteins ; genetics ; metabolism ; Phosphorylation ; Protein-Tyrosine Kinases ; genetics ; metabolism

Objective To observe the effect of Ginkgo biloba extract(GBE)on depression like behavior in post stroke depression(PSD)model rats,and explore the mechanism of regulating Toll like receptor 4/nuclear factor-κ B(TLR4/NF-κB)pathway to inhibit neuroinflammation.Methods Rats were randomly divided into 6 groups,sham,cerebral ischemia,PSD,paroxetine,low-dose Ginkgo biloba extract(GBE-L)and high-dose Ginkgo biloba extract(GBE-H)groups,10 rats in each group.Except for the sham group,middle cerebral artery occlusion(MCAO)was performed to prepare a left focal cerebral ischemia model.Except for the sham group and cerebral ischemia group,other groups were subjected to chronic unpredictable mild stress(CUMS)to establish PSD rat model for 8 weeks.After 4 weeks of CUMS,the paroxetine group,GBE-L,and GBE-H were treated with paroxetine 5 mg·kg-1,GBE 50 mg·kg-1,and GBE 100 mg·kg-1,respectively.The sham group,cerebral ischemia group,and PSD group were treated with the same volume of 0.9％NaCl and continuously administered by gavage for 28 d.After 4 weeks and 8 weeks of CUMS,the body weight and sugar preference test were measured.Levels of serum tumor necrosis factor-α(TNF-α),interleukin-1 β(IL-1 β)and levels of norepinephrine(NE),serotonin(5-HT),and dopamine(DA)in the cerebral cortex were measured by enzyme-linked immunosorbent assay(ELISA).The mRNA levels of Tlr4,Nfkb1,and nuclear factor κ B-kinase subunit β inhibitory factor(Ikbkb)in the hippocampus of rats were detected by polymerase chain reaction.The protein levels of NF-κB,nuclear factor κB inhibitory protein α(IKBα)and phosphorylation nuclear factor κB inhibitory protein α(p-IKB)in hippocampal tissue were detected by Western blot.Results The body weights of rats in the sham group,cerebral ischemia group,PSD group,paroxetine group,GBE-L group and GBE-H group were(427.10±6.36),(403.10±7.37),(310.10±9.71),(355.00±4.03),(347.90±9.88)and(391.90±5.07)g;sugar preference rate were(93.93±1.78)％,(91.57±1.03)％,(54.72±7.34)％,(88.35±4.36)％,(63.55±12.73)％and(81.04±4.31)％;the levels of NE in the cerebral cortex were(1 951.14±52.86),(1 827.27±23.63),(1 662.12±35.92),(2 033.58±72.28),(1 887.31±33.07)and(2 175.00±42.54)pg·mL-1;the levels of 5-HT in the cerebral cortex were(237.07±8.86),(226.15±10.27),(214.51±3.46),(297.13±5.79),(274.14±7.63)and(285.34±8.72)ng·mL-1;the levels of DA in the cerebral cortex were(1 531.11±47.26),(1 209.89±58.09),(1 143.15±36.31),(1 812.67±51.28),(1 651.56±31.82)and(1 853.33±20.42)pg·mL-1.Compared with the PSD group,GBE significantly increased the body weight of rats(P＜0.01)and increased the preference rate of sugar water in rats,showing the antidepressant like behavioral.GBE significantly reduced the levels of serum TNF-α,IL-1 β(all P＜0.01),increased the levels of NE,5-HT,and DA in the cerebral cortex(all P＜0.01),down regulate the mRNA levels of Tlr4,Nfkb1 and Ikbkb(P＜0.05,P＜0.01),reduced the expression of NF-κB(P＜0.01),and reduced the phosphorylation of IKBα(P＜0.01).Conclusion Ginkgo biloba extract can improve depression-like behavior in PSD model rats,and has antidepressant effect.Its mechanism is related to the inhibition of TLR4/NF-κB pathway,thus reducing neuroinflammation.

OBJECTIVETo observe the effects of different concentrations of PPAR gamma agonist rosiglitazone on hypoxia/reoxygenation-induced oxidative stress, cell viability and apoptosis in rat cardiac myocytes.

METHODSCultured rat cardiac myocytes were divided into 5 groups, namely group I (normal group), group II (20 micromo/L ROS group), group III (I/R group), group IV (I/R+20 micromo/L ROS group), and group V (I/R+80 micromo/L ROS group). Group IV and group V were treated with rosiglitazone 12 h before hypoxia/reoxygenation. The changes in cell morphology were observed under optical and transmission electron microscopy, and levels of malondialdehyde (MDA), superoxide dismutase (SOD) activity, and lactate dehydrogenase (LDH) content were determined after the treatment. MTT assay was performed to assess the cell viability and flow cytometry was used to analyze the cell apoptosis.

RESULTSHypoxia/reoxygenation resulted in significantly increased MDA and LDH contents and apoptosis of the cardiac myocytes (P<0.05), but lowered SOD activity and the cell viability (P<0.05). The MDA and LDH contents and apoptotic rate were significantly lower but SOD content and cell vitality significantly higher in groups IV and V than in group III (P<0.05). Group V showed significantly lower MDA and LDH contents and apoptotic rate but higher but SOD content and cell vitality than group IV (P<0.05). Electron microscopy revealed obvious apoptotic changes in group III, and only mild changes were found in group V.

CONCLUSIONRosiglitazone can significantly reduce hypoxia/reoxygenation-induced oxidative stress in cardiac myocytes, improve the cell viability and dose-dependently reduce the apoptotic rate of the cardiac myocytes.

Animals ; Apoptosis ; drug effects ; Cell Hypoxia ; Cell Survival ; drug effects ; Immunohistochemistry ; L-Lactate Dehydrogenase ; metabolism ; Malondialdehyde ; metabolism ; Microscopy, Electron, Transmission ; Myocytes, Cardiac ; cytology ; drug effects ; metabolism ; ultrastructure ; Oxidative Stress ; drug effects ; Oxygen ; metabolism ; PPAR gamma ; agonists ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism ; Thiazolidinediones ; pharmacology

OBJECTIVETo investigate the effectiveness of reconstruction of immunological functions of T cells on the degree of metastases of mouse hepatocarcinoma and the mechanisms of their functioning.

METHODSThe T cell model of immunological functions in Balb/c nu/nu mice was established and the effectiveness of the model was evaluated. The mice were divided into 4 groups. The immunological functions of T cells in experiment groups of Balb/c nu/nu mice were reconstructed. Metastases of the cancer in lymph nodes in each group were examined histologically. The formation time and growth rate of the tumors were calculated. The expression of MHCI and II of the tumor cell line and the difference of expression of immune associated gene were detected by Th1-Th2-Th3 gene array.

RESULTSThe ratio of CD3, CD4, CD8 and CD4/CD8 in the reconstructed group was higher than that in the control group. The average formation time was 7.7+/-0.6 days in Balb/c nu/nu mice and 11.5+/-1.3 days in Balb/c mice. The extent of metastases of the experiment group was lower than that of the control group (P < 0.05). The expression of MHCI of the high metastasis cell line was lower than that of the low metastasis cell line (P < 0.05). The expressions of Th1/Th2 associated genes in lymphocytes of high metastasis mice were lower than those of the low metastasis mice.

CONCLUSIONReconstruction of the immunological function of T cells can influence the metastasis of mouse hepatocarcinoma. The alteration of MHC molecule and low expression of Th1/Th2 correlated genes in lymphocytes may be a factor influencing the metastasis of liver cancer.

Animals ; CD4-CD8 Ratio ; Carcinoma, Hepatocellular ; immunology ; pathology ; Liver Neoplasms ; immunology ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; T-Lymphocytes ; immunology ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Tumor Cells, Cultured

OBJECTIVESTo study the inhibition effect of Daltepartin Sodium (low molecular weight heparins) on human hepatocellular carcinoma (HCC) in nude mice.

METHODSMetastatic model of HCC was established in nude mice. The model mice were randomly divided into 4 groups; they were the control group (saline solution), chemotherapy group (fluorouracil and Cis-dichlorodiamine platinum), Daltepartin Sodium group (Daltepartin Sodium), combined treatment group (Daltepartin Sodium and chemotherapy). Tumor sizes, tumor inhibition rates, tumor metastases, intratumoral microvessel density (MVD), CD31 and AFP were evaluated.

RESULTSIn comparison with the control and the chemotherapy group, the tumor sizes of the Daltepartin Sodium and the combined treatment group were significantly smaller; the tumor inhibitor rates were 0% versus 93.6%, 76.7%, 78.0%; MVD were 20.7+/-6.8 versus 18.2+/-2.6, 4.8+/-1.8 and 6.5+/-2.4; CD31 were 31.8+/-5.7 versus 25.5+/-5.1, 21.6+/-4.8 and 19.6+/-2.4; The incidence of liver metastasis was 80%, versus 70%, 20% and 10%; lung metastasis was 70% versus 60%, 20% and 10%; the peritoneal metastasis was 90% versus 60%, 30%and 30%. AFP were 121.8 ng/ml+/-31.4 ng/ml versus 21.5 ng/ml+/-13.3 ng/ml, 75.6 ng/ml+/-29.7 ng/ml and 55.8 ng/ml+/-38.0 mg/ml. Inhibiting effects of growth and metastasis of HCC in chemotherapy, Daltepartin Sodium and combined treatment groups were significantly different from those of the control group (F=9.191, P < 0.01), Daltepartin Sodium inhibited the angiogensis in the tumors more effectively than that in the control and chemotherapy groups (F=4.937, P < 0.01).

CONCLUSIONDaltepartin Sodium can inhibit tumor growth and metastasis by inhibiting tumor angiogenesis in our nude mice HCC model.

Angiogenesis Inhibitors ; therapeutic use ; Animals ; Carcinoma, Hepatocellular ; drug therapy ; pathology ; Heparin, Low-Molecular-Weight ; therapeutic use ; Liver Neoplasms, Experimental ; drug therapy ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Metastasis ; prevention & control ; Neoplasm Transplantation ; Random Allocation