Objective: To investigate the influence of polysaccharide of Spirulina platensis(PSP) on inhibition activity of immobilized phycocyanin (PC) against proliferation of pancreatic cancer cells SW1990. Methods: PSP was added to the wells of cell culture polystyrene which was immobilized with PC, and SW1990 cells were cultured on this kind of biomaterial in serum free medium. Results: When the concentration of PSP was 15 mg/L, the inhibition activity was mainly caused by PSP, and when the concentrations were 10, 17.5 and 25 mg/L, PSP was cooperative with immobilized PC. The best immobilized amount of PC was 0.02 mg/well, which could inhibit 86% SW1990 cells. Conclusion: PSP can enhance the inhibition activity against pancreatic cancer cells SW1990 from immobilized PC.

Poly-N-isopropylacrylamide (PNIPAAm) is a new kind of intelligent material. It shows favorable thermo sensitivity because of the structure of hydrophilic acrylamino and hydrophobic isopropyl. PNIPAAm also shows good biocompatibility and non-toxicity. All the characters as above make it an ideal extra cellular matrix material for tissue engineering. This paper reviews its application in tissue engineering.
Acrylamides ; chemistry ; Acrylic Resins ; Animals ; Biocompatible Materials ; Hot Temperature ; Humans ; Polymers ; chemistry ; Tissue Engineering ; Tissue Scaffolds

OBJECTlVE To estabIish a simpIe,sensitive and quick method for determination of B7011 in rat pIasma. METHODS The method of protein precipitation with methanoI was used for pre-treatment of pIasma sampIes determined by Iiquid chromatography mass spectrometer. The Iinear reIa-tionship,intra-batch and inter-batch precision,specificity,matrix effect,recovery rate,the accuracy and stabiIity of the pIasma sampIes were vaIidated. The concentration of B7011 in pIasma was determined by LC-mS/ mS foIIowing a singIe intravenous injection of B7011 0.5 mg·kg-1 to rats. RESULTS The Iinear range of B7011 was 30-20 000 μg·L-1 ,the Iower Iimit of quantification was 30 μg·L-1 in pIasma,the in-tra-batch precision of 60,1000,16 000 and 10 000 ng·mL-1 was 5.61% -13.31%,2.31% -8.35%, 2.02%-9.47% and 4.0%-15.0% respectiveIy,and inter-batch precision was 10.05%,2.55%,3.75% and 8.58% respectiveIy. The recovery of 60,1000,and 16 000 μg·L-1 was 114.12%,109.2% and 101.06%respectiveIy. The average peak concentrations were 8373.28 and 8564.59 μg·L-1 ,the mean AUC was 98 400 and 104 000 μg·L-1·h and the t1/ 2z for B7011 was 41.7 and 63.6 h in bIood of maIe and femaIe rats, respectiveIy. CONCLUSlON The estabIished method is sensitive, fast and simpIe and concentration of B7011 in pIasma is determined by LC-mS/ mS foIIowing a singIe intravenous injection of B7011 0.5 mg·kg-1 to rats. It can satisfy the requirements of pharmacokinetic and toxicokinetic studies.

The bioactive protein-phycocyanin and all the proteins of Spirulina Platensis were isolated and purified. Photo-reactive proteins were synthesized by coupling the proteins with (N-(4-azidobenzoyloxy)succinimide) and were spread onto the 24-well cell culture polystyrene plate. Then the coated surface was exposed to ultraviolet irradiation for chemical fixation of proteins via the conversion of the phenylazido group to the highly reactive phenyl-nitrene which spontaneously formed covalent bonds with neighboring hydrocarbons. On these proteins-immobilized polystyrene plates, the liver cancer cells 7402 were cultured under the serum-free conditions, and the inhibition activity on proliferation of liver cancer cells was investigated and analyzed.
Animals ; Bacterial Proteins ; chemistry ; pharmacology ; Biocompatible Materials ; pharmacology ; Cell Division ; drug effects ; Cyanobacteria ; chemistry ; Liver Neoplasms ; pathology ; Photochemistry ; Polystyrenes ; chemistry ; Spirulina ; Succinimides ; chemistry ; Tumor Cells, Cultured

The aim of this study was to evaluate the changes in the expression of Bax, Bcl-2 and P53 when HeLa cells were induced by free cytokines or co-immobilized cytokines. The cells were induced for 24 hrs,72 hrs, 120 hrs and 168 hrs. Then, the expression of Bax, Bcl-2 and P53 was observed by immunohistochemistry. The average optic density of reaction products was tested by image analysis. Lastly, data were analyzed statistically. After the HeLa cells were induced for 120 hrs, the average optic denisity of Bcl-2 was much decreased. However, the average optic Bax was much increased. The average optic density of P53 also increased with the increase of time. The results suggest that HeLa apoptosis was induced by tumor necrosis factor-a and interferon -gamma, and the increasing expression of P53 may induce the expression of Bax and prevent the expression of Bcl-2, via the mitochondrial induction of cell apoptosis.
Apoptosis ; drug effects ; HeLa Cells ; Humans ; Immobilized Proteins ; pharmacology ; Interferon-gamma ; pharmacology ; Photochemistry ; methods ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology ; Tumor Suppressor Protein p53 ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

This study inquired into the mechanisms of co-immobilized cytokines and free cytokines-induced apoptosis on HeLa cells. With the use of photochemical fixed method, TNF-alpha/IFN-gamma were co-immobilized on a 24-well polystyrene culture plate. HeLa cells were stained with fluorescent probe JC-1 to detect the changes of mitochondrial membrane potential (deltapsim), and then were examined by flow cytometry. The results showed that co-immobilized cytokines could induce the apoptosis of HeLa cells in a dose-independent manner. When treated with low-dose of co-immobilized cytokines (20ng/ml), the mitochondrial membrane potential (deltapsim) of HeLa cells continually decreased in 6 days. These indicate that low dose co-immobilized cytokines have a long-term of apoptosis-inducing effect on HeLa cells. We assume that there is close relationship between the mitochondrial membrane potential decrease and the apoptosis of HeLa cells.
Apoptosis ; drug effects ; Dose-Response Relationship, Drug ; HeLa Cells ; Humans ; Immobilized Proteins ; pharmacology ; Interferon-gamma ; pharmacology ; Membrane Potential, Mitochondrial ; drug effects ; Mitochondrial Membranes ; drug effects ; physiology ; Tumor Necrosis Factor-alpha ; pharmacology

This study was aimed to examine the expression of apoptosis-associated gene Fas in HeLa cell, explore the effects of the co-immobilized cytokines (tumor necrosis factor-alpha and interferon-gamma), and probe the potential mechanism of action. The preparation and application of the research couple IFN-gamma and TNF-alpha to the polystyrene cell culture plate were performed using the Photo-immobilization method, with different doses (20 ng/well and 200 ng/well) and synthesized optical active material. HeLa cells were treated with cytokines for two dose and 1, 3, 6 days. The result showed that the free cytokines induced HeLa apoptosis quickly, yet the HeLa apoptosis induced by co-immobilized cytokines had longer effect.
Apoptosis ; drug effects ; genetics ; Drug Synergism ; HeLa Cells ; Humans ; Immobilized Proteins ; chemistry ; pharmacology ; Interferon-gamma ; chemistry ; pharmacology ; Tumor Necrosis Factor-alpha ; chemistry ; pharmacology ; Up-Regulation ; fas Receptor ; metabolism

Objective To differentiate the renal angiomyolipoma with minimal fat (mf-RAML) from clear cell renal cell carcinoma (ccRCC) by Logistic regression analysis and CT quantitative analysis.Methods 25 patients with mf-RAML and 62 patients with ccRCC were collected and performed with enhanced CT scan.The CT signs,CT attenuation value and CT relative attenuation value were calculated and recorded.Results Pseudocapsule and enhancement pattern (OR=0.061,7.445,P=0.005,0.034) were the independent variables of Logistic regression equation.There were statistical difference between the unenhanced muscle％,CT relative attenuation value in renal cortical phase,CT attenuation value and CT relative attenuation value in renal medulla phase and CT relative attenuation value of the mf-RAML and those of the ccRCC (P =0.000-0.040).In addition,the two diseases had low positive correlation with the unenhanced muscle％ and CT attenuation value and CT relative attenuation value in renal medulla phase (r =0.221-0.343,P =0.001-0.040),and had low to middle negative correlation with CT relative attenuation value in renal cortical phase (r =-0.512,-0.225,P =0.000,0.036).The areas under the ROC curve of unenhanced muscle％ and CT relative attenuation value in renal medulla phase were relatively high (0.665-0.753).Conclusion Pseudocapsule and enhancement pattern were independent factors in differentiating mf-RAML from ccRCC.The CT attenuation value of the mf-RAML was similar to the erector spinae muscle.The enhanced degree of mf-RAML in renal cortical phase was lower than that of the ccRCC,while the reduced degree of the ccRCC was higher than that of the mf-RAML in renal medulla phase.The unenhanced muscle％ and the CT relative attenuation value in renal medulla phase are of great significance in diagnosing mf-RAML and ccRCC,especially the unenhanced muscle％.

IFN-gamma and TNF-alpha were co-coupled to the polystyrene cell culture plate by the photo-immobilization method. To investigate the synergistic effect of IFN-gamma and TNF-alpha on the HeLa cells, HeLa cells were treated with co-coupled cytokine or non-coupled cytokine in a time course in this study. The morphology detection, cell cycle analysis by flow cytometry, phosphatidyl serine analysis and capase-3 activity detection demonstrated that the two kinds of treatments both induce HeLa cells apoptosis. Non-coupled cytokine worked more quickly while co-coupled cytokine kept more permanent effect. The caspase-3 activity assay indicated that the caspase-3 activity of HeLa cells treated with non-coupled cytokine is higher than that of HeLa cells treated with co-coupled cytokine. This may imply that co-coupled cytokine not only induces the caspase-dependent pathway, but also induces the caspase-independent pathway.
Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Drug Synergism ; HeLa Cells ; Humans ; Immobilized Proteins ; pharmacology ; Interferon-gamma ; pharmacology ; Tumor Necrosis Factor-alpha ; pharmacology

BACKGROUND: The symptoms of allergic rhinitis (AR) greatly affect the quality of life (QoL) in the patients with AR. The correlations of nasal response to leukotriene D4 (LTD4) and histamine nasal provocation with health related QoL in AR are not clear. OBJECTIVE: To evaluate the correlations of nasal response to LTD4 and histamine nasal challenge with QoL in AR. METHODS: Patients randomly underwent LTD4 and histamine nasal challenge tests, completed the rhinoconjunctivitis quality of life questionnaire (RQoLQ), and rating the symptom severity score (total symptom score 4, TSS4) in the previous week. The correlations between nasal challenge tests induced nasal responses and QoL in RQoLQ were analyzed. RESULTS: A total of 25 eligible AR patients enrolled and finished both LTD4 and histamine nasal challenge and completed the questionnaire of RQoLQ. Histamine nasal challenge induced sneezing, increased nasal resistant were correlated with most of the dimensions (general, practical, nasal, eye problems, and quality of sleep, p < 0.05), while LTD4 nasal challenge induced sneeze, increased nasal resistant only correlated with nasal and ocular problems. On the contrary, the severity of the sneeze assessed by TSS4, was not correlated with QoL, while the severity of rhinorrhea, congestion, and nasal pruritus were correlated with nasal and practical problems, and nasal congestion was also correlated with ocular problems (r = 0.60, p = 0.01). CONCLUSION: LTD4 and histamine nasal challenge induced nasal responses were correlated with different clinical symptoms severity and QoL, which can be used as a good diagnosis and evaluation methods for the management of AR.
Diagnosis ; Estrogens, Conjugated (USP) ; Histamine ; Humans ; Leukotriene D4 ; Pruritus ; Quality of Life ; Rhinitis, Allergic ; Sneezing