AIM: To study the function of N-copine on cell level by investigating the effect of N-copine antisense on the cultured cortical neuron of mouse embryos. METHODS: Neuronal cultures were prepared from cerebral cortices of 16-day old mouse (BALB/C) embryos. N-copine expression was blocked by using antisense oligonucleotides. The effect of inhibition of N-copine expression on the cultured neuronal viability and development was observed. RESULTS: With oligonucleotides treatment for 72 h, the length of neuronal axon and the area of neurosoma in antisense group were decreased markedly as compared with those in control group. The number of trypan-blue-stained neurons and the LDH activity of the culture media were increased significantly as compared with those in control groups. CONCLUSIONS: Inhibition of N-copine expression affects the development of cortical neuron cultured in vitro, and induces the neuron demaged severely. The results suggest that N-copine is a functional protein in neurons, and it may play an important role in the regeneration of nervous system and preventing neuronal degeneration.

Objective To investigate the effects of Xuebijing injectio (Chinese herb preparation) on intestinal function and inflammatory responses in severe burn patients. Method Thirty-two patients with comparable severity in burn injury were randomly divided into Xuebijing injectio treatment group (n = 16) and control group (n = 16). Patients in both groups received routine burn therapy, while those in Xuebijing treatment group additionally received Xuebijing injectio 100mL in intravenous drip twice a day for 7 days. Before the treatment and on the 3rd and the 7th day after the treatment, blood concentrations of diamine oxidase (DAO), lipopolysaccharide (LPS) ,tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6) were determined in both groups. Analysis was made by SPSS 12.0 software. Results The plasma levels of DAO, LPS, TNF-α and IL-6 were decreased in both groups after the treatment. However, the plasma levels of DAO, LPS, TNF-α and IL-6 in the Xuebijing treatment group were significantly lower than those in control group on the 3rd and 7th day after the treatment (P＜ 0.05).Conclusions Xuebijing injectio could protect intestinal function, decrease the plasma level of endotoxin and lessen zhe inflammatory responses in severe burn patients.

Alpha-glycerophosphate oxidase (alpha-GPO) from Enterococcus casseli flavus was successfully isolated and purified by using polyethylene glycol (PEG)/(NH4)2SO4 aqueous two-phase system (ATPS). The results showed that the chosen PEG/(NH4)2SO4 ATPS could be affected by PEG molecular weight, pH, concentration of PEG and (NH4)2SO4, and inorganic salt as well as additional amount of crude enzyme. After evaluating these influencing factors, the final optimum purification strategy was formed by 16.5% (m/m) PEG2000, 13.2% (m/m) (NH4)2SO4, pH 7.5 and 30% (m/m) additive crude enzyme, respectively. The NaCl was a negative influencing factor which would lead to lower purification fold and activity recovery. These conditions eventually resulted in the activity recovery of 89% (m/m), distribution coefficient of 1.2 and purification fold of 7.0.
Ammonium Sulfate ; chemistry ; Glycerolphosphate Dehydrogenase ; chemistry ; isolation & purification ; Molecular Weight ; Polyethylene Glycols ; chemistry ; Water

Objective To study the affect and the related molecular mechanism of glycogen synthase kinase-3β in the proliferation of osteosarcomaand its value in the target therapy of osteosarcoma.Methods The expression level of p-GSK-3β(Ser9)and GSK-3β were detected in human osteoblast cell and osteosarcoma cells by western blot.Observe the effect of GSK-3β inhibitors and siRNA interference on the GSK-3β regulate osteosarcoma cells using apoptosis protein chip.Evaluate the valueof GSK-3β target therapy on osteosarcoma in vivo.Results The expression level of p-GSK-3β (Ser9)was lower in osteosarcoma cells.LiCL,GSK inhibitor Ⅸ,siRNA knockdown could inhibit the cell viability and up-regulated the apoptosis-related protein cleaved-caspase3.The results of the protein array showed that downstream proteins of NF-κB downregulated significantly.The results were validated by western blot,while the downregulation of p-Iκ-Bα and nuclear NF-κB p65 were also observed after LiCL treatment.Inhibition of GSK-3β by either LiCl or specific siRNA resulted in a significant reduction of NF-κB luciferase reporter activity.Furthermore,the NF-κB luciferase reporter activity was significantly increased in CA cell lines,but not in KD cell lines.By contrast,NF-κB-luciferase reporter activity was significantly decreased in stably GSK-3β knockdown cells.GSK3β inhibitor LiCL and shRNA knock down demonstrated a strong cytotoxicity effect on osteosarcoma cells in vivo.Conclusion GSK-3β is in the state of relative active in osteosarcoma in osteosarcoma and important in cell proliferation.GSK-3β regulates cell survival partially through the NF-κB pathway.It is a promising therapeutic target in osteosarcoma.

Objective To investigate the expression level and clinical significance of melanoma antigen gene A (MAGEA) in osteosarcoma patients. Methods Compare gene expression profiles in osteosarcoma cell lines and osteoblasts with gene microarrays. Validation of differentially expressed genes was carried out by real-time polymerase chain reaction analysis. Corresponding protein levels were measures by Western blot analysis in osteosarcoma cell lines and by immunohistochemistry in osteosarcoma tissues. The staining intensity of immuno-histochemistry was correlated with clinical outcome , and its prognostic significance was analyzed. Results Sev-eral genes belonging to MAGEA increased significantly in all osteosarcoma cell lines and tumor tissue , but not in normal osteoblast cell. Patients with MAGEA expression has higher risk of lung metastasis (relative risk 2.79, 95% confidence interval, 1.12-6.93; P = 0.028) and lower five-year survival rates (39.6% ± 8.4% vs. 80% ± 8.9%, P = 0.01) compared with patients without MAGEA expression. Conclusions The expression of MAGEA increased in osteosarcoma , which inversely correlating with outcome of osteosarcoma patients.

BACKGROUND AND OBJECTIVEPrecious studies proven that aberrant tyrosine phosphorylation has linked with cancer. The aim of this work is to study the expression and significance of SHP2 in non-small cell lung cancer (NSCLC) through tissue microarray technique and immunohistochemical method.

METHODSEighty NSCLC specimens were constructed into tissue microarray and performed using immunohistochemistry.

RESULTSThe total positive rates of SHP2 were 70.7% (56/80) in NSCLC, 72.5% (29/40) in squamous cell carcinoma and 75% (27/40) in adenocarcinoma, which was not significant difference in sex, age, the size of tumor, histology, clinical stages and differentiation (P > 0.05), the positive rates of SHP2 were significantly higher in the cases with lymphnode metastasis than those without (P < 0.05).

CONCLUSIONThe expression rate of SHP2 is high and closely correlated to lymphnode metastasis in NSCLC, which implies the occurrence and development of lung cancer maybe related to SHP2, and SHP2 maybe a new marker and therapeutic targets for lung cancer.

Adenocarcinoma ; metabolism ; pathology ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; pathology ; Male ; Middle Aged ; Protein Tyrosine Phosphatase, Non-Receptor Type 11 ; metabolism ; Tissue Array Analysis

Objective:To design and synthesize photosensitizers with different substituents and to identify its physicochemical characteritics and photodynamic effect on tumor cells.Methods:Two kinds of BODIPY photosensitizers BPOI and BPCI were synthesized through condensation reaction between aldehyde and reactive hydrogen of pyrrole,followed with electrophilic substitution reaction.Physicochemical properties were characterized by 1H NMR,FT-IR and UV-visible absorption spectra and fluorescence emission spectra.The ability to produce reactive oxygen species was detected by BPDF and DCFH-DA.Photodynamic therapy effect on rat glioma C6 cells in vitro was determined by MTT method.Results:Two kinds of BODIPY photosensitizers BPOI and BPCI were successfully synthesized with different substituents,which were confirmed by 1H NMR,FT-IR.Both materials had low toxicity and could be readily taken up by tumor cells.The ability of synthesized photosensitizers to produce reactive oxygen species was strongly influenced by solvent polarity when the substituent belongs to electron-donating group,while no effect was found when the substituent belongs to electron-withdrawing group.Conclusion:Photosensitizer BPOI with electron-donating substituent produces reactive oxygen species with a slow rate in a highly polar environment,while the ability to produce reactive oxgen is greatly improved in a low polarity environment,which is expected to be used for environmental-selective photodynamic therapy in tumor cells.

OBJECTIVETo detect the expression of SCUBE3 in human osteosarcoma cell lines and surgical specimens of osteosarcomas and investigate its association with the patients' prognosis.

METHODSThe expression of SCUBE3 protein was detected in 5 osteosarcoma cell lines using Western blotting. CCK8 assay was used to assess the effect of SCUBE3 suppression mediated by two specific small interfering RNAs (siRNAs) on the proliferation of U2OS osteosarcoma cells, and the cell cycle changes were detected using flow cytometry. Immunohistochemistry was performed to detect the expression of SCUBE3 in 60 osteosarcoma tissues, and Kaplan-Meier method was performed for survival analysis of the patients.

RESULTSCompared with osteoblast hFOB1.19 cells, the osteosarcoma cell lines all showed significantly higher expressions of SCUBE3. In U2OS cells, suppression of SCUBE3 expression by siRNA significantly inhibited the cell proliferation (P<0.05). Kaplan-Meier survival analysis indicated that patients with high SCUBE3 expression had worse prognosis than those with low SCUBE3 expression (P=0.036).

CONCLUSIONSCUBE3 is up-regulated in the 5 osteosarcoma cell lines and in primary osteosarcoma tissues to promote the proliferation of osteosarcoma cells. A high SCUBE3 expression in osteosarcoma tissues is associated with a poor prognosis of the patients, suggesting that SCUBE3 can serve as a potential therapeutic target for osteosarcoma.

Bone Neoplasms ; metabolism ; pathology ; Calcium-Binding Proteins ; metabolism ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Flow Cytometry ; Humans ; Immunohistochemistry ; Kaplan-Meier Estimate ; Osteosarcoma ; metabolism ; pathology ; Prognosis ; RNA, Small Interfering ; Up-Regulation

Tumor tissues are composed of tumor cells and complicate microenvironment. Tumor associated macrophages (TAMs) as an important component in tumor microenvironment, play fundamental roles in tumor progression, metastasis and microenvironment regulation. Recently, studies have found that nanotechnology, as an emerging platform, provides unique potential for cancer imaging and therapy. With the nanotechnology, TAMs imaging presents direct evidence for cancer development, progression, and the effectiveness of cancer treatments; it also can regulate the immunosuppression of tumor microenvironment and improve therapeutic efficiency through TAMs targeted killing or phenotypic transformation. In this article, we illustrate the function of TAMs and review the latest development in nano-carriers and their applications in tumor associated macrophage targeting cancer imaging and therapy.

OBJECTIVETo detect the expression of CXCL14 in human osteosarcoma cell lines and tissues and investigate its association with the prognosis of the patients.

METHODSRT-PCR, enzyme-linked immunosorbent assay (ELISA) and real-time PCR were used to detect the expression of CXCL14 in 4 osteosarcoma cell lines and in 40 pairs of osteosarcoma tissues and adjacent muscular tissues. CCK8 assay and colony formation assay was used to assess the effect of CXCL14 suppression mediated by two specific siRNAs on the proliferation of U2OS osteosarcoma cells. Immunohistochemistry was performed to detect the expression of CXCL14 in 58 osteosarcoma tissues, and Kaplan-Meier method and log-rank test were performed for survival analysis of the patients.

RESULTSSignificant up-regulation of CXCL14 expression was found in the osteosarcoma cell lines and in osteosarcoma tissues compared with the adjacent muscles (P<0.01). In U2OS cell, suppression of CXCL14 expression by siRNA significantly inhibited the cell proliferation (P<0.01) and colony formation rate (P<0.05). Kaplan-Meier survival analysis indicated that patients with high CXCL14 expression had worse prognosis than those with low CXCL14 expression (P=0.02).

CONCLUSIONCXCL14 is up-regulated in both osteosarcoma cell lines and primary osteosarcoma tissues to promote the proliferation of osteosarcoma cells. A high CXCL14 expression in osteosarcoma tissues is associated with a poor prognosis, suggesting the that CXCL14 serve as a potential therapeutic target for osteosarcoma.

Bone Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Chemokines, CXC ; metabolism ; Humans ; Osteosarcoma ; metabolism ; pathology ; Prognosis