Objective To investigate the effect of cyclosporine A(CsA)on viral protein syn- thesis and hepatitis B virus(HBV)DNA replication in vitro.Methods The HBV DNA transfected cell line HepG2.2.15 was treated with different concentration of CsA(0.6-20.0?g/ml)for 4 days. Hepatitis B surface antigen(HBsAg)and hepatitis B e antigen(HBeAg)in supernatant were detected by enzyme-linked immunosorbentassay(ELISA);intracellular hepatitis B core antigen(HBcAg)mR- NA and HBV DNA were analyzed by RT-PCR and slot blot hybridization,respectively;the phospho- rylation at tyrosine acid position 402 of PyK2 kinase(PyK2 Y402)was detected by Western blot.Re- sults CsA could suppress the expression of HBsAg and HBeAg,and inhibit the HBV DNA replica- tion in a dose-dependent manner.The suppression rate of HBsAg and HBeAg under the action of CsA at the concentration of 10.0?g/ml for 4 days was 49.7% and 34.3%,respective;similar effect was observed on HBV DNA replication,HBV DNA was only 34.9% of the control at the concentration of 10.0?g/ml of CsA.The phosphorylation level of PyK2 Y402 was declined under the action of CsA at the concentration of 2.0?g/ml.Conclusions CsA can inhibit the expression of HBsAg,HBeAg and HBV DNA replication in the HepG2.2.15 cell line in a dose-dependent manner.Suppression of the phosphorylation level of PyK2 Y402 maybe involved in the mechanism of the inhibitory activity of CsA on HBV replication.

OBJECTIVETo make silica coating through sol-gel process, and to evaluate the wettability of dental alumina ceramic with or without coating.

METHODSSilica coating was prepared with colloidal silica sol on In-Ceram alumina ceramic surface which had been treated with air particle abrasion. Coating gel after heat treatment was observed with atomic force microscope (AFM), and was analyzed by infrared spectrum (IR) with gel without sintered as control. Contact angles of oleic acid to be finished, sandblasted and coated ceramic surface of were measured.

RESULTSAFM pictures showed that some parts of nano-particles in coating gel conglomerated after heat treatment. It can be seen from the IR picture that bending vibration absorption kurtosis of Si-OH also vanished after heat treatment. Among contact angles of three treated surface, the ones on polished surface were the biggest (P = 0.000, P = 0.000), and sandblasting+silica coating surface the smallest (P = 0.000, P = 0.003).

CONCLUSIONSilica coating can be made with sol-gel process successfully. Heat treatment may reinforce Si-O-Si net structure of coating gel. Wettability of dental alumina ceramic with silica coating is higher than with sandblasting and polishing.

Aluminum Oxide ; Ceramics ; Dental Bonding ; Dental Porcelain ; Materials Testing ; Silicon Dioxide ; Surface Properties

Field experiments were conducted in farmers' rice fields in 2001 and 2002 to study the effects of nitrogen (N) management strategies on N use efficiency in recovery (RE), agronomy (AE) and physiology (PE) and redistribution of dry matter accumulation (DMA) and nitrogen accumulation (NA) in two typical rice cultivars in Jinhua, Zhejiang Province. This study aimed mainly at identifying the possible causes of poor fertilizer N use efficiency (NUE) of rice in Zhejiang by comparing farmers' fertilizer practice (FFP) with advanced site-specific nutrient management (SSNM) and real-time N management (RTNM). The results showed that compared to FFP, SSNM and RTNM reduced DMA and NA before panicle initiation and increased DMA and NA at post-flowering. There is no significant difference between SSNM and FFP in post-flowering dry matter redistribution (post-DMR) and post-flowering nitrogen redistribution (post-NR). These results suggest that high input rate of fertilizer N and improper fertilizer N timing are the main factors causing low NUE of irrigated rice in the farmer's routine practice of Zhejiang. With SSNM, about 15% of the current total N input in direct-seeding early rice and 45% in single rice could be reduced without yield loss in Zhejiang, China.
Crops, Agricultural ; metabolism ; Fertilizers ; Nitrogen ; metabolism ; Oryza ; metabolism

Objective To explore effect of cyclooxygenase-2 ( COX-2) expression on cell viability , proliferation, apoptosis and other biological behavior of human colon cancer cell line HT-29.Methods The expression of COX-2 was examed by Western blot and real-time PCR after siRNA COX-2, empty body was transferred into human color-ectal cancer cell line HT-29 by liposome .Cell viability was measured by CCK-8 method .Cell apoptosis and cell cy-cle was measured by flow cytometry .Cell migration and invasion was detected by scratch test and Transwell method , respectively.The expression of Bcl-2, Bax, matrix metalloproteinase 2 (MMP2) was measured by real-time PCR. The level of protein kinase B ( AKT) phosphorylation was measured by western blot .Results Compared with the control group and empty vector group, the expression of COX-2 protein and mRNA was decreased ( P<0.01) , cell viability was reduced ( P<0.01) , cell apoptotic rate was increased ( P<0.01) , cell cycle was arrested in G phase ( P<0.01 ) , cell migration and invasion ability was decreased ( P<0.01 ) , the expression of Bcl-2, MMP2 mRNA and p-AKT protein was down-regulated (P<0.01), the expression of Bax mRNA was up-regulated in COX-2 inhibition group ( P<0.01 ) .Conclusions Down-regulation of COX-2 expression can significantly inhibit HT-29 cell proliferation , migration and invasion , and induce cell apoptosis , which may be related to the down-reg-ulation of AKT phosphorylation .

OBJECTIVETo establish a bioluminescent MDA-MB-231 cell line which can stably express luciferase and green fluorescent protein to allow bioluminescent imaging in nude mouse models bearing human triple-negative breast cancer xenografts.

METHODSThe lentivirus carrying luc2, eGFP and neo fusion genes were packaged in 293T cells via calcium phosphate co-precipitation. Human triple-negative breast cancer cell line MDA-MB-231 was infected by the lentivirus, and the positive cell clones were tested for eGFP and luc2 expressions by fluorescence microscopy and Xenogen IVIS200 bioluminescent imaging system, respectively. MTT assay, transwell invasion assay and wound healing assay were performed to evaluate the changes in the proliferation, invasion and migration abilities of the infected cells. The cells were then orthotopically implanted into the right second mammary fat pat of female BALB/c nude mice. The tumor growth was monitored by the in vivo imaging system every week, and the tumor tissues were harvested to evaluate the in vivo stability and tumorigenicity of the modified cells using cryosection and HE staining.

RESULTSThe lentivirus-infected MDA-MB-231cells could stably express luc2 and eGFP, and the luciferase activity reached 9689 phontons/s/per cell. No significant changes occurred in the biological activities of the lentivirus-infected MDA-MB-231 cells. We successfully established the nude mouse model bearing orthotopically implanted human triple-negative breast cancer cells.

CONCLUSIONThe modified MDA-MB-231 cell line can be detected sensitively at the primary implantation site and distant metastasis site in nude mice, which provides a convenient and sensitive platform for the research of metastatic mechanism and new antitumor drugs of human triple-negative breast cancer. The combination of eGFP and luc2 is superior to single reporter gene.

Animals ; Brain Neoplasms ; secondary ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Disease Models, Animal ; Female ; Genes, Reporter ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Humans ; Lentivirus ; genetics ; metabolism ; Luciferases ; biosynthesis ; genetics ; Luminescent Measurements ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; Neoplasm Transplantation ; Receptor, ErbB-2 ; metabolism ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism

OBJECTIVETo isolate and identify the cancer stem cells from primary human ovarian cancer tissues.

METHODSFresh tumor tissues from five cases of pathologically diagnosed ovarian cancers were taken, minced and then digested with collagenase and hyaluronidase to obtain single cell suspension. The erythrocytes were removed with ACK Lysis buffer. The suspensions were sorted by magnetic activated cell sorting (MACS) using CD133-binding microbeads. Then the sorted CD133(+) cells were verified by flow cytometry. The cells were cultured in serum-free medium supplemented with EGF, bFGF, insulin and BSA, and grew into spheroids. Immunofluorescence, differentiation and tumor formation tests of the cells were performed to characterize the properties of cancer stem cells.

RESULTSThe ovarian cancer stem cells were successfully isolated from primary human ovarian tumors, which formed typical spheroids in serum-free medium and had stronger ability of tumorigenesis. The results of related experiments verified that CD133 positive cells owned the properties of cancer stem cells.

CONCLUSIONSThe ovarian cancer stem cells presenting the characteristics of stemness in vitro and in vivo, have been successfully isolated from primary human ovarian tumor tissues by MACS. The isolated ovarian cancer stem cells could be used in future researches of their biological functions.

AC133 Antigen ; Animals ; Antigens, CD ; metabolism ; Cell Differentiation ; Cell Separation ; methods ; Female ; Flow Cytometry ; methods ; Glycoproteins ; metabolism ; Humans ; Immunomagnetic Separation ; methods ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Neoplasm Transplantation ; Neoplastic Stem Cells ; metabolism ; pathology ; Ovarian Neoplasms ; metabolism ; pathology ; Peptides ; metabolism

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

This study was to establish an iron overload bone marrow (BM) model by co-culturing the mononuclear cells from BM with iron, and investigate its hematopoiesis changes. The iron overload model was set up by adding different concentration of ferric citrate (FAC) into the mononuclear cells from BM and culturing for different time, and the model was confirmed by detecting labile iron pool (LIP). Then the apoptosis of hematopoietic cells, ability of hematopoietic colony forming (CFU-E, BFU-E, CFU-GM and CFU-mix) and percentage of the CD34(+) cells of the BM cells all were determined. The changes of these indexes were tested after the iron-overloaded BM was treated with deferasirox (DFO). The results showed that after BM cells were cultured with FAC at different concentrations for different time, the LIP increased in time-and concentration-dependent manners. The intracellular LIP reached maximum level when cultured at 400 µmol/L of FAC for 24 hours. The detection of BM cell hematopoietic function found that the apoptotic rate of the FAC-treated cells (24.8 ± 2.99%) increased significantly, as compared with normal control (8.9 ± 0.96%)(p < 0.01). The ability of hematopoietic colony forming in FAC-treated cells decreased markedly, as compared with normal control (p < 0.05). The percentage of CD34(+) cells of FAC-treated cells (0.39 ± 0.07%) also decreased significantly, as compared with normal control (0.91 ± 0.12%)(p < 0.01). And these changes could be alleviated by adding DFO. It is concluded that the iron-overloaded model has been set by adding iron into the mononuclear cells from BM in vitro, and the hematopoietic function of iron-overloaded BM is deficient. These changes can be alleviated by removing the excess iron from the BM cells through treating with DFO. These findings would be helpful to further study the mechanism of iron-overload on the hematopoiesis of BM and also useful to find the way to treat iron-overload patients with hematopoietic disorders.
Bone Marrow Cells ; cytology ; Cells, Cultured ; Hematopoiesis ; Hematopoietic Stem Cells ; cytology ; Humans ; Iron ; metabolism ; Iron Overload

OBJECTIVETo investigate the prognostic factors for non-small cell lung cancer(NSCLC)in patients under 40 years of age.

METHODSThe clinicopathological data of 148 young patients with NSCLC were retrospectively analyzed. Kaplan-Meier and Cox regression analyses were used to analyze the relationship between prognostic factors and survival time.

RESULTSThe patients were followed-up for 6 - 148 months, and the follow-up rate was 100%. In the whole group, 122 patients died and 26 cases were surviving. The 1-, 3- and 5-year survival rates were 54.7%, 10.4% and 5.6%, respectively. The median survival time (MST) was 14.7 months. Kaplan-Meier analysis showed that Karnofsky performance status (KPS), clinical stage, treatment modality and serum CEA were related with prognosis (P < 0.05). Multivariate analysis indicated that KPS, clinical stage, treatment modality and serum CEA were independent prognostic factors (P < 0.05).

CONCLUSIONSKPS, CEA, clinical stage and treatment modalities are independent prognostic factors in young NSCLC patients.

Adolescent ; Adult ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoembryonic Antigen ; blood ; Carcinoma, Non-Small-Cell Lung ; blood ; drug therapy ; pathology ; radiotherapy ; surgery ; Combined Modality Therapy ; Female ; Follow-Up Studies ; Humans ; Karnofsky Performance Status ; Lung Neoplasms ; blood ; drug therapy ; pathology ; radiotherapy ; surgery ; Male ; Neoplasm Staging ; Pneumonectomy ; methods ; Proportional Hazards Models ; Retrospective Studies ; Survival Rate ; Young Adult

OBJECTIVE: To determine the family member's depression status and its influencing factors in the patient with mental disorder caused by cerebral trauma. METHODS: The self-designed information questionnaire and self-rating depression scale (SDS) were used to investigate 245 family members' depression conditions. On the basis of SDS score index, all cases were divided into depression group and non-depression group. The data were analyzed by the univariate analysis and multiple factors regression analysis. RESULTS: In the 245 family members, 117 family members had different degrees of depressive symptoms including 68 with mild depression, 42 with moderate depression, and 7 with severe depression. In the multiple factors regression analysis, genetic relationship, personality characteristics and payment method were the main influencing factors of the depression status. CONCLUSION Much attention should be paid to the family member's depression condition of the patient with mental disorder caused by cerebral trauma through specific psychological intervention.
Accidents, Traffic ; Adult ; Brain Injuries/complications* ; Depression/psychology* ; Family/psychology* ; Family Relations ; Female ; Humans ; Male ; Mental Disorders/psychology* ; Middle Aged ; Regression Analysis ; Severity of Illness Index ; Socioeconomic Factors ; Stress Disorders, Post-Traumatic/psychology* ; Surveys and Questionnaires