Protective Effect of Cortex Phellodendri Extract on Myocardial Injury in Rats / 中国药师

Objective:To investigate the protective effects and possible mechanisms of cortex phellodendri water extract and etha -nol extract on the myocardial injury induced by pituitrin and isoproterenol hydrochloride in rats .Methods:SD rats as the experimental animals were randomly divided into the normal control group , model group , compound Danshen tablets group , phellodendron water ex-tract group and phellodendron ethanol extract group .Pituitrin and isopropyl adrenaline hydrochloride were used to establish the myocar-dial injury model in rats.The serum CK, LDH activity, myocardial tissue SOD activity and MDA content were detected and compared . Results:Compared with those in the normal control group , the serum LDH activity , CK activity and MDA content were significantly in-creased , and the SOD activity in cardiac muscle and myocardial tissue was significantly decreased in the pituitrin -established myocardi-al injury model group (P<0.01).In the isopropyl adrenaline hydrochloride-established myocardial injury model group , the MDA con-tent in myocardial tissue was obviously increased , and the SOD activity in myocardial tissue was decreased obviously (P<0.01).The serum LDH activity, CK activity and MDA content were significantly decreased , and the SOD activity in cardiac muscle and myocardial tissue was increased significantly in all drug-taken groups when compared with those in the pituitrin-established myocardial injury model group, and the differences were statistically significant (P<0.05 or P<0.01).The MDA content in myocardial tissue was significant-ly reduced , and the SOD activity was increased significantly in all drug-taken groups when compared with those in the isopropyl adrena-line hydrochloride-established myocardial injury model group , and the differences were statistically significant (P<0.05 or P<0.01). Conclusion:Cortex phellodendri extract has certain protective effect on myocardial injury induced by pituitrin and isopropyl adrenaline hydrochloride in rats .

The anti-tumor effect of Coix stalk alcohol extraction on H22 tumor-bearing mice / 天津医药

Objective To study the anti-tumor effects of alcohol extraction of Coix stalk objects on H22 tumor-bearing mice. Methods The animal model of tumor bearing mice with H22 ascitic tumor cells was established. Eighty-four model mice were randomly and equally divided into Coix stalk extract groups 1-5 (10, 8, 6, 4 and 2 g/kg), model control group and cyclophosphamide group. Mice were treated orally with Coix stalk alcohol extraction solution (10, 8, 6, 4 and 2 g/kg), cyclophosphamide 0.02 g/kg and normal saline once a day for 8 days for Coix stalk extract group, cyclophosphamide group and model control group. The mouse activity, the size and the appearance of time of abdominal swelling, and changes of hair, feeding and drinking water quantity were observed in groups of mice. The solid tumor mass was measured in H22 tumor-bearing mice. The tumor inhibitory rate, liver index, spleen index and thymus index were calculated. Results The axillary tumor muster was found first in model control group with the fastest growth, reduced independent activity, decreased appetite and dim in hair color, followed by the Coix stalk extract group 1 and group 2. The last was Coix stalk extract group 5 and cyclophosphamide group. The solid tumor mass were (0.47±0.18), (0.37± 0.13), (0.34±0.10), (0.30±0.11) and (0.28±0.09) mg for Coix stalk alcohol extract groups 1-5, which were significantly lower than those of model control group (0.60 mg±0.21 mg, F=5.700,P<0.05). The tumor inhibition rates were 21.67%, 38.33%, 43.33%, 50.00%, 53.33%and 60.00%in Coix stalk extract groups 1-5 and cyclophosphamide group. The liver index, spleen index and thymus index were lower in cyclophosphamide group and Coix stalk alcohol extract groups than those of model control group (except for the spleen index of Coix stalk extract group 1). The liver index was lower in Coix stalk ethanol extract groups than that of cyclophosphamide group. There were no significant differences in the spleen index, thymus index between Coix stalk ethanol extract groups and cyclophosphamide group. Conclusion Coix stalk alcohol extract has inhibitory effects on the tumor and liver damage in H22 mice.

Expression of GRP78 in the radiated survival subclones of nasopharyngeal carcinoma C666-1 cells / 中国医师杂志

Objective To investigate radio-sensitivity and expression of GRP78 protein in the survival subclones of nasopharyngeal carcinoma (NPC) C666-1 cells.Methods NPC C666-1 cells were first irradiated with X-ray at a large dose of 8Gy.Three survival subclones were selected and named as C666-1-R1, C666-1-R2, and C666-1-R3.The radio-sensitivity was analyzed for the radiated survival subclones and their parent control C666-1 cells with Methyl Thiazolyl Tetrazolium assay(MTT) and Trypan blue dye methods.The expression of GRP78 was analyzed for three survival subclones and control C666-1 with Western blot.Results After 6 Gy irradiation, the cell survival rate of three subclones was higher than that of the control cells, especially a significant difference for C666-1-R2 cells (P ＜ 0.05), which suggested a radioresistance in C666-1-R2 cells.Moreover, GRP78 expression in each subclone was significantly higher than that of parent C666-1 cells (P ＜ 0.05).Conclusions The irradiated-survival subclone C666-1-R2 was radio-resistant.GRP78 was overexpressed in the irradiated-survival subclones.GRP78 might be an ideal target for treatment of a nasopharyngeal carcinoma.

Expression of HSP70/HSP27 protein in residual lesion after NPC radiotherapy / 中南大学学报(医学版)

OBJECTIVE@#To analyze HSP70/HSP27 protein expression in the nasopharyngeal carcinoma (NPC) primary tissues and the residual lesion, and to explore its predictive value.@*METHODS@#Immunohistochemical experiment was performed to detect the expression of HSP70 and HSP27 in 58 NPC primary tissues: 28 were in the experimental group with the local residual lesion and 30 in the control group without recurring and metastasis within 5 years by conventional radiotherapy.@*RESULTS@#The positive expression of HSP70 and HSP27 in the experimental group was 92.9%(26/28) and 53.6%(15/28), while that in the control group was 53.3%(16/30) and 53.3%(16/30),respectively. There was significant difference in the 2 groups. The common positive expression of HSP70 and HSP27 between the 2 groups had significant difference, 50.0% (14/28) in the experimental group and 16.7% (5/30) in the control group, respectively. There was no significant difference in the negative ratio of HSP70 and HSP27 common expression between the 2 groups, 3.6% (1/28) in the experimental group and 10.0% (3/30) in the control group, respectively.@*CONCLUSION@#HSP70 may have an important role in the radioresistance of NPC, and may predict the residual lesion after radiotherapy.

Screening of EGFR-regulated Secreted Proteins in Human NPC Cell Line CNE2 / 生物化学与生物物理进展

In order to screen EGFR-regulated secreted proteins in human nasopharyngeal carcinoma(NPC), and to reveal the role and mechanism of epidermal growth factor receptor(EGFR) in the pathogenesis of NPC. NPC cell line CNE2 cells were cultured in serum-free medium and stimulated by transforming growth factor-? (TGF-?) for 24 h in experimental group. Control CNE2 cells were cultured at the same condition but without TGF-? stimulation. The culture medium of control and experimental cells was desalted and concentrated through ultrafiltration to prepared the total secreted proteins. Two-dimensional gel electrophoresis (2-DE) was used to separate the secreted proteins of control and experimental cells, PDQuest software was applied to analyze 2-DE images, and the differential protein spots between the control and experimental cells were identified by desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The 2-DE patterns of the secreted proteins of TGF-? stimulated and un-stimulated CNE2 cells were established, 22 differential proteins spots between the two groups of cells were found, and 8 non-redundant proteins were identified with MALDI-TOF-MS, the functions of which were involved in invasion, metastasis, apoptosis and proliferation of cancer cells. The data will be valuable for further to study the role and mechanism of EGFR in the pathogenesis of NPC.

Differential Proteomic Analysis of Sera From Lung Squamous Carcinoma Patients and Healthy Individuals / 生物化学与生物物理进展

To screen for serum biomarkers for lung squamous carcinoma, two-dimensional gel electrophoresis (2-DE) was performed to separate serum proteins from healthy individuals and stage 1 lung squamous carcinoma(LSC) patients, respectively. PDquest software was used to analyze 2-DE images, and the differential serum protein spots between the healthy individuals and LSC patients were identified by ESI-Q-TOF MS/MS. Then Western blot and immunohistochemistry were used to detect the expression levels of haptoglobin-2(HP-2), one of the differential proteins, in the sera and tumor tissues in the patients with LSC, respectively. 2-DE maps of serum proteins from healthy individuals and stage 1 LSC patients were established. Ten differential serum protein spots were detected, four proteins of which were identified by MS/MS. Western blot showed that the serum level of HP-2 in the LSC patients was significantly higher than that in healthy individuals, but was not associated with LSC staging. Immunohistochemistry showed that the expression level of HP-2 in the LSC tissues was significantly higher than that in the normal bronchial epithelial tissues adjacent to tumors. The results indicated that serum HP-2 protein is a candidate biomarker for LSC, and might be useful for diagnosis of LSC. Up-regulation of HP-2 in the LSC tissues may contribute to the high serum level of HP-2 in the patients.

Proteomic comparison of two-dimensional gel electrophoresis profiles from human lung squamous carcinoma and normal bronchial epithelial tissues / 基因组蛋白质组与生物信息学报·英文版

Differential proteome profiles of human lung squamous carcinoma tissue compared to paired tumor-adjacent normal bronchial epithelial tissue were established and analyzed by means of immobilized pH gradient-based two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). The results showed that well-resolved, reproducible 2-DE patterns of human lung squamous carcinoma and adjacent normal bronchial epithelial tissues were obtained under the condition of 0.75-mg protein-load. The average deviation of spot position was 0.733+/-0.101 mm in IEF direction, and 0.925+/-0.207 mm in SDS-PAGE direction. For tumor tissue, a total of 1241+/-88 spots were detected, 987+/-65 spots were matched with an average matching rate of 79.5%. For control, a total of 1190+/-72 spots were detected, and 875+/-48 spots were matched with an average matching rate of 73.5%. A total of 864+/-34 spots were matched between tumors and controls. Forty-three differential proteins were characterized: some proteins were related to oncogenes, and others involved in the regulation of cell cycle and signal transduction. It is suggested that the differential proteomic approach is valuable for mass identification of differentially expressed proteins involved in lung carcinogenesis. These data will be used to establish human lung cancer proteome database to further study human lung squamous carcinoma.

EGFR Activates AP-1 Through ERK-1/2 Signaling Pathway in the Rat Gastric Mucosal Cells / 中国医师杂志

Objective To explore the signaling pathway of AP-1 activated by epidermal growth foctor receptor(EGFR) in the rat gastric mucosal cells. Methods TGF-? stimulated freshly isolated rat gastric mucosal cells,and Western blot and EMSA detected the activation of ERK-1/2 signaling pathway. Results The exposure of the mucosal cells to 1 nmol/L TGF-? for 30 min significantly induced transcriptional activity of AP-1. This induction was associated with a concomitant activation of MEK and ERK-1/2. TGF-?-induced activation of AP-1 in gastric mucosal cells could be totally abrogated by either PD153035, a specific inhibitor of EGFR tyrosine kinase, or PD98059, a specific inhibitor of MEK. Conclusions Our findings suggest that EGFR AP-1 activates by EPK-1/2 signaling pathway in the rat gastric mucosal cells.