Objective: To observe the curative effects of rh-endostatin combined with DP regimen in treating patients with advanced esophageal cancer and analyze the correlation of CT perfusion (CTP) parameters and the expression of vascular endothelial growth factor (VEGF). Methods: Twenty patients with esophageal cancer confirmed pathologically were randomly divided into combined treatment (rh-endostatin+DP regimen) group and single chemotherapy group, 10 patients in each group, respectively. All patients were given conventional CT examination and CTP imaging for primary tumor. The level of VEGF, the size of tumor and CTP parameters (BF, BV, PS and MTT) before treatment and after 2 cycles of treatment were determined for the comparison and the correlation between CTP parameters and VEGF expression was analyzed. Results: the therapeutic effect of rh-endostatin+DP regimen group was superior to single chemotherapy group. VEGF level after treatment in rh-endostatin+DP regimen group was obviously lower than single chemotherapy group (P<0.01). The expression of VEGF had positive correlation with BF and BV but negative correlation with MTT. Compared with treatment before for rh-endostatin+DP regimen group, BF, BV and PS decreased while MTT increased after treatment (P<0.05). However, there were no significant differences between treatment before and after treatment in single chemotherapy (P>0.05). Conclusions: Rh-endostatin can down-regulate the expression of VEGF in esophageal cancer, change the state of hypertransfusion and high permeability of tumor vessels and had the better curative effect and slighter adverse reactions when combined with chemotherapy.
Potential antioxidant effect of processed ginseng (sun ginseng, SG) on oxidative stress generated by tert-butyl hydroperoxide (t-BHP) was investigated in HepG2 cells. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and lactate dehydrogenase (LDH) leakage test demonstrated that SG dose-dependently prevents a loss of cell viability against t-BHP-induced oxidative stress. Also, SG treatment dose-dependently relieved the increment of activities of hepatic enzymes, such as aspartate aminotrasferase and alanine aminotransferase, and lipid peroxidation mediated by t-BHP treatment in HepG2 cells. SG increased the gene expression of antioxidant enzymes such as superoxide dismutase, catalase, and glutathione peroxidase. However, high dose of SG treatment caused decrease in mRNA level of glutathione peroxidase as compared to low dosage of SG-treated cells. The gene expression of glutathione reductase was found to be slightly increased by SG treatment. In addition, SG extract attributed its hepaprotective effect by inducing the mRNA level of bcl-2 and bcl-xL but reducing that of bax. But, the gene expression of bad showed no significant change in SG-treated HepG2 cells. These findings suggest that SG has hepatoprotective effect by showing reduction of LDH release, activities of hepatic enzymes and lipid peroxidation and regulating the gene expression of antioxidant enzymes and apoptosis-related molecules against oxdative stress caused by t-BHP in HepG2 cells.
Background: Hepatocellular carcinoma is one of the most common cancers worldwide. Its prevalence is increasing in many countries. Plant products can be used to protect against cancer due to natural anticancer and chemopreventive constituents. Strobilanthes crispus is one of plants with potential chemopreventive ability. Objective: This study aimed to evaluate the anticancer effects of Strobilanthes crispus juice on hepatocellular carcinoma cells. Materials and Methods: MTT assays, flow cytometry, comet assays and the reverse transcription-polymerase chain reaction (RT-PCR) were used to determine the effects of juice on DNA damage and cancer cell numbers. Results: This juice induced apoptosis after exposure of the HepG2 cell line for 72 h. High percentages of apoptotic cell death and DNA damage were seen at the juice concentrations above 0.1%. It was found that the juice was not toxic for normal cells. In addition, juice exposure increased the expression level of c-myc gene and reduced the expression level of c-fos and c-erbB2 genes in HepG2 cells. The cytotoxic effects of juice on abnormal cells were in dose dependent. Conclusions: It was concluded that the Strobilanthes crispus juice may have chemopreventive effects on hepatocellular carcinoma cells.
Kim, Mi-Jin;Kim, Sang-Hyuk;Lee, Jae-Hyung;Seo, Jin-Ho;Lee, Jong-Hwan;Kim, Jong-Hyun;Kim, Yeon-Hee;Nam, Soo-Wan
Journal of Microbiology and Biotechnology
/
v.18
no.12
/
pp.1938-1944
/
2008
The procpb gene encoding human procarboxypeptidase B (proCPB, GeneBank access code AJ224866) was cloned and its Pichia expression plasmid, $pPIC9{\alpha}$/hproCPB (9.2 kb), was constructed, in which procpb was under the control of the AOXl promoter and connected to the downstream of the mating factor ${\alpha}$-1 ($MF{\alpha}1$) signal sequence. The plasmid was linearized by digestion with Sacl, and integrated into the genome of P. pastoris strain GS115. By culturing of Pichia transformant on methanol medium, the human proCPB was successfully expressed and secreted into the culture supernatant. Moreover, Western blot analysis of the extracellular proteins showed proCPB bands clearly at a molecular mass of 45 kDa, confirming the expression of proCPB with its right size. The CPB activity reached about 3.5 U/ml and 12.7 U/ml in the flask and fermentor batch cultures of Pichia transformant, respectively. No CPB enzyme activity was found in the intracellular fraction. When the fed-batch cultivation was performed with methanol and glycerol mixture as a feeding medium, the extracellular CPB activity was increased to 42.0 U/ml, which corresponds to a 3.3-fold higher level of CPB activity than that of batch culture. The $K_m$ and $k_{cat}$ values of recombinant human CPB enzyme for hippuryl-$_L$-Arg as a substrate were estimated to be 0.16 mM and $11.93\;sec^{-1}$, respectively.
The influence of high temperature on mammary cell turnover during the dry period is still unclear. The objective of this study was to investigate mammary cell turnover and p53 protein expression in the mammary tissue under high temperature conditions. Mammary gland biopsy samples from 8 dairy cows were obtained at 7, 25, 40, and 53 d during the dry period in summer or spring (n = 4, each season). Cell cycle, cell turnover, and p53 protein expression were analyzed by flow cytometry. During the dry period in summer, the percentage of mammary epithelial cells in the G0/G1 phase was the highest, but those in the S and G2/M phases were lower. However, the proportion of cells in the different stages of the cell cycle was not significantly different among the different biopsy time points, except in the G2/M phase. Under different temperature conditions, the cells were significantly different in their apoptotic rate and proliferation index; moreover, the tendencies of these indicators to change significantly differed. In general, the samples under high temperature conditions showed significantly lower apoptotic rates and proliferation indices. Under high temperature conditions, the apoptotic rate and proliferation index were the lowest (2.17% and 3.26%, respectively) at day 40, and the highest at day 53 (3.67% and 4.61%, respectively). However, under normal temperature conditions, the values of these indicators were the lowest (7.60% and 5.54%, respectively) at day 7, and almost the highest at day 25 (12.85% and 6.47%, respectively). Moreover, p53 protein expression was significantly higher under high temperature conditions than under normal temperature conditions, except at day 25. The level of p53 protein was the lowest (13.10%) under high temperature conditions at day 25, but was the highest (26.07%) under normal temperature conditions. Our findings suggest that high temperature delayed the G2/M phase of the cell cycle and the cell turnover rate, but remarkably increased p53 protein expression. Thus, the results indicate that high temperature extends the recovery period of mammary epithelial cells.
Chicken egg ovalbumin is a non-inhibitory member of the serpin (serine protease inhibitors) family whose members share a common tertiary fold. In the present study, we succeeded in high-level production of a disulfide-free form of refolded recombinant ovalbumin. Conformational characterization of the recombinant ovalbumim revealed that it is well-folded, following two-state unfolding transition with the midpoint of transition at 4.7 M at $25^{\circ}C$. This value is very close to that of the reduced form of authentic ovalbumin. The recombinant ovalbumin can serve as a model molecule of non-inhibitory serpins in comparative studies with inhibitory members of the serpin family.
Many fashion brands provide sophisticated customization services with advanced technology. Previous studies have revealed that a high level of customization aided by technology positively influences purchase intention. However, since fashion products are society-approved symbols, brand prestige and consumer propensity are expected to affect purchase behavior of customized products. This study aims to identify the purchase decision mechanism of customized products by identifying the relationships among self-expression, brand prestige, narcissistic disposition, and purchase intention. An online survey was conducted on 220 Korean women in their 20s and 30s with customization experience. The results of the data analysis showed that the affluent customization area and narcissistic disposition positively affected purchase intention. Surprisingly, when brand prestige was high and narcissistic disposition was low, the more affluent the customization area, the higher the purchase intention. However, for higher narcissistic disposition, no significant relationship was found between the customization area and purchase intention. This study provides insights into discriminatory customization strategies based on consumer propensity and the brand prestige level when providing customization services in fashion brands.
The purpose of this study was to find out the Bcl-2 (B-cell leukemia/lymphoma-2), Bax, and caspase-3(cysteine-aspartic proteases-3) protein expression in soleus and EDL muscle according to treadmill exercise intensity in 60 week-old SD rats. The SD rats were randomly divided into four groups (n=10 in each group): control (CON), low intensity exercise (LE), moderate intensity exercise (ME), and high intensity exercise (HE). The exercise was given to the rats for 8 wk, 5 day/wk. The animals underwent treadmill exercise at intensities of 30 min at 8 m/min for the LE group, 15 min at 16 m/min for the ME group, and 9 min at 24 m/min for the HE group. The results were as follows: the expression of Bcl-2 protein was lowest in the HE group and the expression of Bax protein was highest in the HE group. The expression of caspase-3 (cleaved form) protein was observed in the HE group. For the different types of muscle fiber, Bcl-2 protein expression in the soleus muscle was decreased in all groups. Bax protein expression in the soleus muscle was increased in the HE group only. Bcl-2 protein expression in the EDL muscle was decreased in the HE group, and Bax protein expression in the EDL muscle was increased in the ME and HE groups. Consequently, the protein expression related to the aged rats shows a difference according to the intensity of exercise. In addition, caspase-3 protein expression appeared in the HE group; however, in all amounts of intensity, DNA fragmentation was not observed. Therefore, apoptosis on skeletal muscles of aged mice can be intervened with optimal exercise. On the other hand, high intensity exercise can potentially accelerate the apoptosis of muscle fiber in aged rats.
Aims: Mammalian target of rapamycin (mTOR) is master regulator of the PI3K/Akt/mTOR pathway and plays an important role in NSCLCs. Here we characterized mRNA and protein expression levels of mTOR and its functional associated molecules including PTEN, IGF-1R and 4EBP1 in surgically resected NSCLCs. Methods: Fifty-four patients with NSCLCs who underwent pulmonary resection were included in current study. mRNA levels of mTOR, PTEN, IGF-1R, and 4EBP1 were evaluated by RT-PCR and protein expression of mTOR, PTEN, and IGF-1R by immunohistochemistry (IHC). Association of expression of the relevant molecules with clinical characteristics, as well as correlations between mTOR and PTEN, 4EBP1 and IGF-1R were also assessed. Results: The results of RT-PCR showed that in NSCLCs, the expression level of mTOR increased, while PTEN, 4EBP1 and IGF-1R decreased. Statistical analysis indicated high IGF-1R expression was correlated with advanced clinical stage (stage III) and PTEN expression was reversely associated with tumor size (P=0.16). The results of IHC showed mTOR positive staining in 51.8% of cases, while IGF-1R positive staining was found in 83.3% and loss of PTEN in 46.3%. Protein expression of mTOR was correlated with its regulators, PTEN and IGF-1R, to some extent. Conclusions: Abnormal activation of mTOR signaling, high expression of IGF-1R, and loss of PTEN were observed in resected NSCLC specimens. The poor expression agreement of mTOR with its regulators, PTEN, and IGF-1R, implied that combination strategy of mTOR inhibitors with other targets hold significant potential for NSCLC treatment.
Objectives : The purpose of this study was to investigate the effects of Jowiseungcheung-tang (JWSCT) extract on the lipid metabolism, anti-oxidation and inflammatory reflex. Methods : Male Sprague-Dawley rats were fed a high fat diet for 8 weeks and were randomly divided into 4 groups (10 mice in each group): control group, 100 mg/kg JWSCT group, 200 mg/kg JWSCT group, 300 mg/kg JWSCT group. The control group was administered 100 mg/kg of water, but the other three groups were administered 100, 200, 300 mg/kg JWSCT extract for 4 weeks. After 4 weeks, we measured lipid level, thiobarbituric acid reactive substance (TBARS), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), catalase (CAT), aspartate aminotransferase (AST), alanine aminotransferase (ALT) and cytokines in plasma and liver. The gene expression level and the ratio of apo-B and apo-E were then investigated by way of reverse transcription -polymerase chain reaction (RT-PCR). Results : In the JWSCT group, compared with the control, free fatty acid, triglyceride, total cholesterol, LDL-cholesterol, TBARS, IL-$1{\beta}$, IL-6 and TNF-${\alpha}$ decreased significantly in plasma and liver. However HDL-cholesterol, IL-10, GSH-Px, SOD and CAT increased. In the JWSCT group, compared with the control, the gene expression level and the ratio of apo-A and apo-E decreased significantly in the RT-PCR analysis. Conclusions : The extract of JWSCT has anti-obesity, anti-inflammatory and anti-oxidant effects.
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