Kim, Jun-Sang;Lee, Sheng-Jin;Kim, Jin-Man;Cho, Moon-June
Radiation Oncology Journal
/
v.26
no.1
/
pp.56-64
/
2008
Purpose: Cathepsin D(CD) is a lysosomal acid proteinase that is related to malignant progression, invasion, and a poor prognosis in several tumors. The aim of this study was to evaluate the prognostic clinical significance of CD and p53 expression in pretreatment biopsy specimens from patients with locally advanced rectal cancer who were treated with preoperative chemoradiation. Materials and Methods: Eighty-nine patients with locally advanced rectal cancer(cT3/T4 or N+) were included in this study. Preoperative chemoradiation consisted of a dose of 50.4 Gy of pelvic radiation and two concurrent cycles of administration of 5-fluorouracil and leucovorin. Surgery was performed six weeks after chemoradiation. CD and p53 expression in pretreatment formalin-fixed paraffin-embedded tumor biopsy specimens were assessed by immunohistochemical staining using a CD and p53 monoclonal antibodies. The threshold value for a positive stain in tumor tissue and stromal cells was 1+ intensity in 10% of the tumors or stromal cells, respectively. Results: Positive CD expression was found in 57(64%) of the tumors and 32(35%) of the stromal cell specimens. There was no association with CD expression of the tumor or stromal cells and patient characteristics. There was a correlation between tumor CD expression with stromal cell CD expression(p=0.01). Overexpression of p53 was not a significant prognostic factor. The 5-year overall survival(OS) and disease-free survival(DFS) rates were not different between tumor CD-negative and positive patient biopsy samples(69% vs. 65%, 60% vs. 61%, respectively). The 5-year OS rates in the tumor-negative/stromal cell-negative, tumor-negative/stromal cell-positive, tumor-positive/stromal cell-negative and tumor-positive/stromal cell-positive biopsy samples were 75%, 28%, 62%, and 73%, respectively. Stromal cell staining only without positive tumor staining demonstrated the worst overall survival prognosis for patients(p=0.013). Conclusion: Overexpression of p53 in rectal biopy tissue was not associated with prognostic significance. In the pretreatment biopsy specimens, an exclusive increase in CD expression in stromal cells without tumor expression was related to poor overall survival in patients with locally advanced rectal cancer treated with preoperative chemoradiation.
Scutellaria radix (SR) has been utilized as a traditional medicine for a variety of diseases including Rheumatoid arthritis and its major flavonoids - baicalein, baicalin, and wogonin - have been reported to exert beneficial health effects, including anti-bacterial, anti-viral, anti-inflammatory, and free-radical scavenging. However, the mechanisms underlying this effect remain poorly understood. The principal objective of this study was to determine the effect of SR on osteoblast and osteoclast cells. SR extract was prepared using 70% ethanol solvent. Osteoblastic MC3T3-E1 cells and osteoclast precursor Raw 264.7 macrophage cells were utilized. SR extract increased MC3T3-E1 cell proliferation and stimulated alkaline phosphatase activity dose-dependently, 152.0% of the control at concentration $1{\mu}g/mL$. Additionally, SR extract ($1{\mu}g/mL$) stimulated Bone nodule formation activity in MC3T3-E1 cells, approximately 223.3% of the control, 20 days after the exposure. In addition, SR extract significantly reduced the number of tartrate-resistant acid phosphatase-positive (TRAP+) multinucleated cells from Raw 264.7 cells. In conclusion, SR extract stimulates the proliferation and bioactivities of boneforming osteoblasts, and inhibits the activities of bone-resorbing osteoclasts to a certain degree.
Jeong, Tae Hyug;Youn, Joo Yeon;Ji, Keunho;Seo, Yong Bae;Kim, Young Tae
Journal of Life Science
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v.24
no.4
/
pp.343-351
/
2014
Phosphoinositide 3-kinase (PI3K) plays a central role in cell signaling and leads to cell proliferation, survival, motility, exocytosis, and cytoskeletal rearrangements, as well as specialized cell responses, superoxide production, and cardiac myocyte growth. PI3K is divided into three classes; type I PI3K is preferentially expressed in leukocytes and activated by ${\beta}{\gamma}$ subunits of heterotrimeric G-proteins. In this study, the cDNAs encoding the $PI3K{\gamma}$ gene were isolated from a brain cDNA library constructed using the flounder (Paralichthys olivaceus). The sequence of the isolated $PI3K{\gamma}$ was 1341 bp, encoding 447 amino acids. The nucleotide sequence of the $PI3K{\gamma}$ gene was analyzed with that of other species, including Oreochromis niloticus and Danio rerio, and it turned out to be well conserved during evolution. The $PI3K{\gamma}$ gene was subcloned into the expression vector pET-44a(+), and expressed in the E. coli BL21 (DE3) codon plus cell. The resulting protein was expressed as a fusion protein of approximately 49 kDa containing a C-terminal six-histidine extension that was derived from the expression vector. The expressed protein was purified to homogeneity by His-tag affinity chromatography and showed enzymatic activity corresponding to $PI3K{\gamma}$. The binding of wortmannin to $PI3K{\gamma}$, as detected by anti-wortmannin antisera, closely followed the inhibition of the kinase activities. The results obtained from this study will provide a wider base of knowledge on the primary structure and characterization of the $PI3K{\gamma}$ at the molecular level.
Kang, Eun Sil;Ham, Sun Ah;Hwang, Jung Seok;Lee, Chang-Kwon;Seo, Han Geuk
Food Science of Animal Resources
/
v.33
no.3
/
pp.411-416
/
2013
This study aimed to examine the mechanisms underlying the effects of Garcinia cambogia extract on the adipogenic differentiation of 3T3-L1 cells and long-chain saturated fatty acid-induced lipotoxicity of HepG2 cells. 3T3-L1 preadipocytes, mouse embryonic fibroblast-adipose like cell line, were treated with MDI solution (0.5 mM IBMX, 1 ${\mu}M$ dexamethasone, 10 ${\mu}g/mL$ insulin) to generate a cellular model of adipocyte differentiation. Using this cellular model, the anti-obesity effect of Garcinia cambogia extract was evaluated. MDI-induced lipid accumulation and expression of adipogenesis-related genes were detected by Oil red O staining, Nile Red staining, and Western blot analysis. Effects Garcinia cambogia extract on palmitate-induced lipotoxicity was also analyzed by MTT assay, LDH release, and DAPI staining in HepG2 cells. Garcinia cambogia extract significantly suppressed the adipogenic differentiation of preadipocytes and intracellular lipid accumulation in the differentiating adipocytes. Garcinia cambogia extract also markedly inhibited the expression of peroxisome proliferator- activated receptor ${\gamma}2$ ($PPAR{\gamma}2$), CCAT/enhancer-binding protein ${\alpha}$ ($C/EBP{\alpha}$), and adipocyte protein aP2 (aP2). In addition, Garcinia cambogia extract significantly attenuated palmitate-induced lipotoxicity in HepG2 cells. Palmitateinduced cellular damage and reactive aldehydes were also significantly reduced in the presence of Garcinia cambogia extract. These findings suggest that the Garcinia cambogia extract inhibits the adipogenic differentiation of 3T3-L1 preadipocytes, probably by regulating the expression of multiple genes associated with adipogenesis such as $PPAR{\gamma}2$, $C/EBP{\alpha}$, aP2, and thereby modulating fatty acid-induced lipotoxicity to reduce cellular injury in hepatocytes.
The present study was evaluated the antibacterial effect of the combination of $Coptidis$$rhizoma$, $Glycyrrhiza$$uralensis$ Fischet, $Schizandra$$chinensis$ and $Corni$$Fructus$(1:1:1) extracts(CGSC10). Furthermore, the effectiveness of CGSC10, sodium chlorate, and the combination of CGSC10 and sodium chlorate(CGSCS10) against $E.$$coli$ O157:H7 infection was studied using ICR female mice. During the incubation period, the dose of 5, 10, and 20% CGSC10 was inhibited the growth of $E.$$coli$ O157:H7 by 34.7, 60.2, and 76.4%, respectively. For 7 days after single challenge with $E.$$coli$ O157:H7, forty female ICR mice were divided into four experimental groups which were administered in drinking water with saline, 10% CGSC10, 15 mM sodium chlorate, and CGSCS10, respectively. On the 3rd day, the number of $E.$$coli$ O157:H7 in mouse feces was significantly decreased by administration of CGSC10, 15 mM sodium chlorate, and CGSCS10 ($p$ < 0.001). On the 7th day-after administration, CGSC10, sodium chlorate, and CGSCS10 were decreased the number of $E.$$coli$ O157:H7 by 27.1, 67.7, and 83.3%, respectively. According to the results of the present study, administration of CGSCS10 to mice can reduce the severity of $E.$$coli$ O157:H7 infection. In addition, it is suggested that CGSCS10 represents a good candidate for the treatment of enteric infections in domestic animals.
The present study was evaluated the antibacterial effect of the combination of Coptidis rhizoma, Lonicerae Flos, and Paeonia japonica (1:1:1) extracts (CLP1000). Also, the effectiveness of CLP1000, dioctahedral smectite (DHS), and the combination of CLP1000 and DHS (CLPS1000) against E. coli O157:H7 infection was studied using ICR female mice. During the incubation period, the dose of 10% and 20% CLP1000 were inhibited the growth of E. coli O157:H7 by 30% and 47%, respectively. For 7 days after single challenge with E. coli O157:H7, forty female ICR mice were divided into four experimental groups which were orally administered with saline, 10% CLP1000, 10% DHS, and 10% CLPS1000, respectively. On the 3rd day, the number of E. coli O157:H7 in mouse feces was significantly decreased by administration of CLP1000 (p < 0.05), DHS (p < 0.05) and CLPS1000 (p < 0.001). On the 7th day, CLP1000 (p < 0.05) and CLPS1000 p < 0.001) administration significantly decreased the number of E. coli O157:H7. According to the results of the present study, administration of CLPS1000 to mice can reduce the severity of E. coli O157:H7 infection. Also, it is suggested that CLPS100 represents a good candidate for the treatment of enteric infections in domestic animals.
Ixeris sonchifolia Hance (Godulbaegi), Oenanthe javanica (Dolminari), Fagopyrum esculentum Moench (Buckwheat>, Hizikia fusiforme (Seaweed Fusiforme) and Zingiber ojficinale Roscoe (Ginger) have been used respectively as one of folk remedies as well as food materials. However, reportedly few studies on their immunomodulating effects have been made, although it has been known from other preceding studies that the ex vivo supplementation of each Ish, OJ, Fem, Hf, Zor water extracts tends to enhance the proliferation of splenocyte in comparison to the control group. This study on the combined immunomodulative effect of water extract mixture of these five food materials (Ish + Oj + Fem +Hf+Zor) lasted covering seven or eight weeks. The old mice (balb/c) was fed ad libitum on chow diet, and the water extract of plant mixture was orally administrated every other day for four weeks at two different concentrations (50 and 500 mg/kg B.W) . After preparing the single cell suspension, the proliferation of splenocyte was determined by MTT (3-[4,5-dimethylthiazol-2-y]-2,5-diphenyl terazolium bromide) assay. The production of cytokine ($IL-{\beta}$, IL-6, and $TNF-{\alpha}$) which was secreted by macrophages stimulated with LPS or not was detected by ELISA assay using the cytokine kit. After the 48 hours of incubation with the mitogen (ConA or LPS) stimulation, the proliferation of the mice splenocyt in the experimental group statisticaly increased at both of two different concentrations in comparison to the control group. The cytokines production was more significantly enhanced at the lower supplementation (50 mg/kg B.W.) group than at the higher concentration (500 mg/kg B.W.). The result of this study may suggest that the supplementation of water extract of plant mixture can regulate and enhance the immune function by increasing the splenocyte proliferation and regulating the cytokine production capacity by the activated macrophages in mice.
This study was carried out to evaluate the antiobesity effects of Crataegus fructus in 3T3-L1 adipocytes and mice fed a high fat diet (high fat 45% cal). The inhibitory effect of methanol extract from Crataegus fructus on lipid accumulation in 3T3-L1 adipocytes was quantified using Oil red O staining. Compared with the control, lipid accumulation was significantly decreased by 10-25% with treatment with Crataegus fructus extract at a concentration of 600-2,000 ug/ml. Three-week old ICR mice (n=24) were randomly divided into four groups (T0: normal diet, T1: high fat diet, T2: high fat diet and 50 ug of Crataegus fructus extract, T3: high fat diet and 100 ug of Crataegus fructus extract) and were fed an experimental diet for 5 weeks. At the end of the experiment, body weight gain in the T1 group (3.9${\pm}$0.24 g) was higher than that in the T0 group (2.56${\pm}$0.14 g), while body weight gain in the T2 (3.02${\pm}$0.25 g) and T3 (2.58${\pm}$0.16 g) groups was significantly reduced as compared with that of the T1 group. Moreover, liver weight in the T1 (4.8${\pm}$0.17 g) and T2 (4.8${\pm}$0.16 g) groups was significantly higher than that of the T0 (4.05${\pm}$0.16 g) and T3 (4.57${\pm}$0.10 g) groups, while kidney weight was significantly lower than that of the T0 and T3 groups (p<0.05). The levels of total cholesterol and triglyceride in serum in the T2 and T3 groups were significantly decreased compared to the T1 group. These results suggest that Crataegus fructus can be used as functional materials in food and medicine.
Objectives : Neuronal changes that result from treadmill exercise for patients with Parkinson's disease(PD) have not been well documented, although some clinical and laboratory reports suggest that regular exercise may produce a neuroprotective effect and restore dopaminergic and motor functions. However, it is not clear if the improvements are due to neuronal alterations within the affected nigrostriatal region or result from a more general effect of exercise on affect areas and motivation. In this study, we demonstrate that motorized treadmill exercise improves the neuronal outcomes in rodent models of PD. Methods : We used a chronic mouse model of parkinsonism, which was induced by injecting male C57BL/6 mice with 10 doses(Every 12 hour) of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (30 mg/kg) and probenecid (20 mg/kg) over 5 days. These mice were able to sustain an exercise training program on a motorized rodent treadmill at a speed of 18 m/min, $0^{\circ}$ of inclination, 40 min/day, 5 days/week for 4 weeks. At the end of exercise training, we extracted the brain and compared their neuronal and neurochemical changes with the control(saline and sedentary) mice groups. Synphilin protein is the substance that manifestly reacts with ${\alpha}$-synuclein. In this study, we used Synphilin as a manifest sign of recovery from neurodegeneration. We analyze the brain stems of the substantia nigra and striatum region using the western blotting technique. Results : There were no expression of synphilin in the saline-induced groups. The addition of MPTP(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) greatly accelerated synphilin expression which meant an aggregation of ${\alpha}$-synuclein. But, the MPTP-induced treadmill exercise group showed significantly lower expression than the MPTP-induced sedentary group. This means treadmill exercise has a definite effect on the decrease of ${\alpha}$-synuclein aggregation. Conclusions : In this study, our results suggest that treadmill exercise promoted the removal of the aggregation of ${\alpha}$-synuclein, resulting in protection against disease development and blocks the apoptotic process in the chronic parkinsonian mice brain with severe neurodegeneration.
As one of the mucous components of Cheonggukjang, traditional fermented soybean paste, $\gamma$-PGA is a natural substance with diverse functions. In this paper, an in-vivo experiment has been performed using NC/Nga mice in order to find out the efficacy of $\gamma$-PGA in human atopic dermatitis. The NC/Nga mice with BMAC-induced atopic dermatitis were administered $\gamma$-PGA (PGA-HM) with 300 kDa and low-molecular $\gamma$-PGA (PGA-LM), respectively. As a result, a significant decrease in clinical skin severity score was detected in the group that was administered PGA-LM. In terms of serum IgE levels, a significant decline was observed in PGA-LM, compared to the control group. The serum IgG1 levels also decreased more in PGA-LM than in the control group. However, no significant difference was observed in both groups. To witness the induction of $CD4^+CD25^+foxp3^+$ Treg cells, mRNA was sampled from the back of PGA-HM- and PGA-LM-administered NC/Nga mice with atopic dermatitis. In terms of the production amount of foxp3 mRNA, which was measured in real-time PCR, the group that was administered PGA-LM was twice as high as the control group. According to a biopsy on the skin on the backs of the mice, the experimental group was also far lower than the control group in terms of epidermis thickness, mast cell infiltration and the number of $CCR3^+$ cells. Therefore, it has been confirmed that the atopic dermatitis symptoms decreased more in the PGA-LM-administered NC/Nga mice than the PGA-HM-administered group by facilitating $CD4^+CD25^+foxp3^+$ Treg cells and suppressing the activity of eosinophils and production of IgE and pro-inflammatory cytokines.
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