• Asian Pacific Journal of Cancer Prevention
• /
• v.14 no.7
• /
• pp.4421-4426
• /
• 2013

Objective: The present study employed 5-aza-2'-deoxycytidine (5-Aza-CdR) to treat non-small cell lung cancer (NSCLC) cell line A549 to investigate the effects on proliferation and expression of the TFPI-2 gene. Methods: Proliferation was assessed by MTT assay after A549 cells were treated with 0, 1, 5, 10 ${\mu}mol/L$ 5-Aza-CdR, a specific demethylating agent, for 24, 48 and 72h. At the last time point cells were also analyzed by flow cytometry (FCM) to identify any change in their cell cycle profiles. Methylation-specific polymerase chain reaction (MSPCR), real time polymerase chain reaction(real-time PCR) and western blotting were carried out to determine TFPI-2 gene methylation status, mRNA expression and protein expression. Results: MTT assay showed that the growth of A549 cells which were treated with 5-Aza-CdR was significantly suppressed as compared with the control group (0 ${\mu}mol/L$ 5-Aza-CdR). After treatment with 0, 1, 5, 10 ${\mu}mol/L$ 5-Aza-CdR for 72h, FCM showed their proportion in G0/G1 was $69.7{\pm}0.99%$, $76.1{\pm}0.83%$, $83.8{\pm}0.35%$, $95.5{\pm}0.55%$ respectively (P<0.05), and the proportion in S was $29.8{\pm}0.43%$, $23.7{\pm}0.96%$, $15.7{\pm}0.75%$, $1.73{\pm}0.45%$, respectively (P<0.05), suggesting 5-Aza-CdR treatment induced G0/G1 phase arrest. MSPCR showed that hypermethylation in the promoter region of TFPI-2 gene was detected in control group (0 ${\mu}mol/L$ 5-Aza-CdR), and demethylation appeared after treatment with 1, 5, 10 ${\mu}mol/L$ 5-Aza-CdR for 72h. Real-time PCR showed that the expression levels of TFPI-2 gene mRNA were $1{\pm}0$, $1.49{\pm}0.14$, $1.86{\pm}0.09$ and $5.80{\pm}0.15$ (P<0.05) respectively. Western blotting analysis showed the relative expression levels of TFPI-2 protein were $0.12{\pm}0.01$, $0.23{\pm}0.02$, $0.31{\pm}0.02$, $0.62{\pm}0.03$ (P<0.05). TFPI-2 protein expression in A549 cells was gradually increased significantly with increase in the 5-Aza-CdR concentration. Conclusions: TFPI-2 gene promoter methylation results in the loss of TFPI-2 mRNA and protein expression in the non-small cell lung cancer cell line A549, and 5-Aza-CdR treatment could induce the demethylation of TFPI-2 gene promoter and restore TFPI-2 gene expression. These findings provide theoretic evidence for clinical treatment of advanced non-small cell lung cancer with the demethylation agent 5-Aza-CdR. TFPI-2 may be one molecular marker for effective treatment of advanced non-small cell lung cancer with 5-Aza-CdR.

• Asian Pacific Journal of Cancer Prevention
• /
• v.15 no.14
• /
• pp.5767-5772
• /
• 2014

Background and Aim: B7-H1, a co-inhibitory molecule of the B7 family, is found aberrantly expressed in ovarian cancer cells and infiltrating macrophage/dendritic-like cells, and plays a critical role in immune evasion by ovarian cancer. IL-12, an inducer of Th1 cell development, exerts immunomodulatory effects on ovarian cancer. However, whether IL-12 regulates B7-H1 expression in human ovarian cancer associated-macrophages has not been clarified. Therefore, we investigated the effects of IL-12 on the expression of B7-H1 in ovarian cancer-associated macrophages and possible mechanisms. Methods: PMA induced THP-1-derived macrophages or human monocyte-derived macrophages were treated with recombinant IL-12 (rIL-12) or infected with adenovirus carrying human IL-12 gene (Ad-IL-12-GFP) for 24 h, then cocultured with the SKOV3 ovarian cancer cell line for another 24 h. Macrophages were collected for real-time PCR and Western blot to detect the expression of B7-H1, and activation of the NF-${\kappa}B$ signaling pathway. Moreover, supernatants were collected to assay for IL-12, IFN-${\gamma}$ and IL-10 by ELISA. In addition, monocyte-derived macrophages treated with IFN-${\gamma}$ were cocultured with SKOV3 and determined for the expression of B7-H1. Furthermore, the expression of B7-H1 in monocyte-derived macrophages was also evaluated after blocking NF-${\kappa}B$ signaling. Results: The expression of B7-H1 was significantly upregulated in monocyte-derived macrophages treated with rIL-12 or Ad-IL-12-GFP compared with the control groups (p<0.05), accompanied by a remarkable upregulation of IFN-${\gamma}$ (p<0.05), a marked downregulation of IL-10 (p<0.05) and activation of NF-${\kappa}B$ signaling. However, the upregulation of B7-H1 was inhibited by blocking the NF-${\kappa}B$ signaling pathway (p<0.05). Expression of B7-H1 was also increased (p<0.05) in monocyte-derived macrophages treated with IFN-${\gamma}$ and cocultured with SKOV3. By contrast, the expression of B7-H1 in THP-1-derived macrophages was significantly decreased when treated in the same way as monocyte-derived macrophages (p<0.05), and IL-10 was also significantly decreased but IFN-${\gamma}$ was almost absent. Conclusions: IL-12 upregulates the expression of B7-H1 in monocyte-derived macrophages, which is possible though inducing the secretion of IFN-${\gamma}$ and further activating the NF-${\kappa}B$ signal pathway. However, IL-12 downregulates the expression of B7-H1 in THP-1-derived macrophages, associated with a lack of IFN-${\gamma}$ and inhibition of expression of IL-10.

• Asian Pacific Journal of Cancer Prevention
• /
• v.14 no.2
• /
• pp.909-913
• /
• 2013

Purpose: The liver is the organ to which colorectal carcinomas (CRCs) most commonly metastasize, and surgical resection has been established as the most effective and potentially curative treatment for CRC with liver metastasis (LM). Therefore, surveillance of LM is vital for improvement of prognosis of CRC patients. In this study, we aimed to explore the potential value of carbohydrate antigen 19-9 (CA 19-9), carcinoembryonic antigen (CEA), and marker enzymes in indicating LM with CRC. Methods: Three groups of eligible patients with metastatic cancers were retrospectively included: CRC patients with LM (CRC-LM) or without LM (CRC-NLM), and non-CRC patients with LM (NCRC-LM). All metastatic lesions were identified by CT or MRI. Data on characteristics of the patients, the primary site, the locations of metastasis, CA 19-9, CEA, and biochemical parameters were collected for analysis. Results: A total of 493 patients were retrospectively included. More alcohol consumption was found in CRC-LM than CRC-NLM. Some biochemical enzymes were found to be significantly higher in groups with LM than without (CRC-LM or NCRC-LM v.s CRC-NLM). Both CEA and CA 19-9 were much higher in CRC-LM than CRC-NLM or NCRC-LM. For CRC patients, CA 19-9, ${\gamma}$-glutamyl transpeptidase, CEA and alcohol consumption were identified as independent factors associated with LM. Conclusion: Our analysis suggested the CA 19-9 might be a potential valuable indicator for LM of CRC in the clinic.

• Journal of Korea Association of Health Promotion
• /
• v.3 no.1
• /
• pp.72-83
• /
• 2005

The present pilot project was executed to recommend a strategy of clonorchiasis control in China. The pilot area of this project was Zhaoyuan, Hailin, and Ningan, Heiloagjiang province. A baseline survey subjecting 4,865 residents in Heilongjiang confirmed Zhaoyuan asa high endemic area and Hailin and Ningan as moderate endemic areas. Six different control strategies were implemented in Zhaoyuan, two were in Hailin, and one was in Ningan. Including the baseline survey and project programs from 2000 to 2004, total 63,274subject-times were examined of their feces for Clonorchiseggs, 26,680 were treated, 10,082 were screened by ELISA, and 6,130 subjects were examined of their liver by sonography. The egg Positive rates in 6 villages of Zhaoyuan were as high as 44.8% 70,0%. Following the protocolof each strategy, the subjected residents were examined of their feces and treated with 25 mg/kg praziquantel, 3 times. Except the control group, all of the villages showed 72.8% to 92.0% reduction of their original egg Positive rates at Zhaoyuan. Mass treatments of all subjected residents in 2001 and 2003 reduced the egg rate from 68.8% to18.7% and 4 annual mass treatments reduced the rate from 44.8% in 2001 to 8.7% in 2004.Selective annual treatments of egg positive subjects reduced the egg rates from 50.8% in2001 to 13.8% in 2004 or from 70.0% in 2001 to 11.6% in 2004, and two treatments in a year reduced the rate from 57.6% in 2001 to 4.6% in 2004. According to repeated treatments, EPG counts decreased remarkably. In moderate endemic areas, the original egg rates were 22.6% and 28.3% in 2001 but were 1.7% and 1.1% after 2 or 3 selective treatments. The present findings of the chemotherapeutic control of clonorchiasis prove that repeated medication is important. The reduction is directly correlated with dose of praziquantel but not with mass or selective treatments. Chemotherapeutic control of reservoirhosts has little effect on reinfection of clonorchiasis because the field along the Songhua-jiang is too wide to be impacted. ELISA confirmed many serologically positive cases to Clonorchisantigen but only a few cases were positive to other antigens (Paragonimus, cysticercus, sparganum). The abdominal soaography visualized intrahepatic bile duct dilatation and periductal echo in 2,002 of 6,070 examined subjects. In addition to these examinations and treatment, health education supplemented tㅗe control activities. The present findings prove clonorchiasis is very widely prevalent and heavily endemic along the rivers in Heiloagjiang. The results suggest that group chemotherapy with praziquantel is effective to reduce endemicity of clonorchiasis. Mass treatment without individual fecal examination is recommended in heavy endemic areas where the egg rate is over 40% while one selective treatment is effective enough in moderate endemic areas.

• Journal of Life Science
• /
• v.24 no.4
• /
• pp.352-360
• /
• 2014

We evaluated the growth performance, biochemical characteristics, and immune responses in weaning pigs given a diet containing MR-1 (0.2%/feed) or antibiotics (0.1%/feed) for 45 days. In vitro study showed that MR-1 has antibacterial activity against a variety of strains of pathogenic bacteria, especially a strain of cattle-derived Escherichia coli K99 (E. coli K99) by agar diffusion assay. In the in vivo model, 0.2% MR-1-given group clearly ameliorated the weight gain and feed efficiency in the growth performance of weaning pigs compared to the basal diet group (p<0.05). Additionally, 0.2% MR-1 induced an elevation in the levels of mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) and showed a similar pattern ($TNF{\alpha}$ and $IFN{\gamma}$ production) to the antibiotic treated pigs. Taken together, we suggest that 0.2% MR-1 makes probiotics an alternative to antibiotics in weaning pigs.

• Archives of Pharmacal Research
• /
• v.28 no.3
• /
• pp.249-268
• /
• 2005

Drug metabolizing enzymes (DMEs) play central roles in the metabolism, elimination and detoxification of xenobiotics and drugs introduced into the human body. Most of the tissues and organs in our body are well equipped with diverse and various DMEs including phase I, phase II metabolizing enzymes and phase III transporters, which are present in abundance either at the basal unstimulated level, and/or are inducible at elevated level after exposure to xenobiotics. Recently, many important advances have been made in the mechanisms that regulate the expression of these drug metabolism genes. Various nuclear receptors including the aryl hydrocarbon receptor (AhR), orphan nuclear receptors, and nuclear factor-erythoroid 2 p45-related factor 2 (Nrf2) have been shown to be the key mediators of drug-induced changes in phase I, phase II metabolizing enzymes as well as phase III transporters involved in efflux mechanisms. For instance, the expression of CYP1 genes can be induced by AhR, which dimerizes with the AhR nuclear translocator (Arnt) , in response to many polycyclic aromatic hydrocarbon (PAHs). Similarly, the steroid family of orphan nuclear receptors, the constitutive androstane receptor (CAR) and pregnane X receptor (PXR), both heterodimerize with the ret-inoid X receptor (RXR), are shown to transcriptionally activate the promoters of CYP2B and CYP3A gene expression by xenobiotics such as phenobarbital-like compounds (CAR) and dexamethasone and rifampin-type of agents (PXR). The peroxisome proliferator activated receptor (PPAR), which is one of the first characterized members of the nuclear hormone receptor, also dimerizes with RXR and has been shown to be activated by lipid lowering agent fib rate-type of compounds leading to transcriptional activation of the promoters on CYP4A gene. CYP7A was recognized as the first target gene of the liver X receptor (LXR), in which the elimination of cholesterol depends on CYP7A. Farnesoid X receptor (FXR) was identified as a bile acid receptor, and its activation results in the inhibition of hepatic acid biosynthesis and increased transport of bile acids from intestinal lumen to the liver, and CYP7A is one of its target genes. The transcriptional activation by these receptors upon binding to the promoters located at the 5-flanking region of these GYP genes generally leads to the induction of their mRNA gene expression. The physiological and the pharmacological implications of common partner of RXR for CAR, PXR, PPAR, LXR and FXR receptors largely remain unknown and are under intense investigations. For the phase II DMEs, phase II gene inducers such as the phenolic compounds butylated hydroxyanisol (BHA), tert-butylhydroquinone (tBHQ), green tea polyphenol (GTP), (-)-epigallocatechin-3-gallate (EGCG) and the isothiocyanates (PEITC, sul­foraphane) generally appear to be electrophiles. They generally possess electrophilic-medi­ated stress response, resulting in the activation of bZIP transcription factors Nrf2 which dimerizes with Mafs and binds to the antioxidant/electrophile response element (ARE/EpRE) promoter, which is located in many phase II DMEs as well as many cellular defensive enzymes such as heme oxygenase-1 (HO-1), with the subsequent induction of the expression of these genes. Phase III transporters, for example, P-glycoprotein (P-gp), multidrug resistance-associated proteins (MRPs), and organic anion transporting polypeptide 2 (OATP2) are expressed in many tissues such as the liver, intestine, kidney, and brain, and play crucial roles in drug absorption, distribution, and excretion. The orphan nuclear receptors PXR and GAR have been shown to be involved in the regulation of these transporters. Along with phase I and phase II enzyme induction, pretreatment with several kinds of inducers has been shown to alter the expression of phase III transporters, and alter the excretion of xenobiotics, which implies that phase III transporters may also be similarly regulated in a coordinated fashion, and provides an important mean to protect the body from xenobiotics insults. It appears that in general, exposure to phase I, phase II and phase III gene inducers may trigger cellular 'stress' response leading to the increase in their gene expression, which ultimately enhance the elimination and clearance of these xenobiotics and/or other 'cellular stresses' including harmful reactive intermediates such as reactive oxygen species (ROS), so that the body will remove the 'stress' expeditiously. Consequently, this homeostatic response of the body plays a central role in the protection of the body against 'environmental' insults such as those elicited by exposure to xenobiotics.

• Journal of Life Science
• /
• v.23 no.7
• /
• pp.893-903
• /
• 2013

The purpose of this study was to evaluate the effects of dietary Flammulina velutipes mycelium (FVM) on physico-chemical properties and nutritional components of chicken meat. Ninety-six broiler chicks (HanHyup No. 3, Korea) were divided into four groups: control (basal diet), T1 (supplemented with 1% FVM), T2 (3%), and T3 (5%). Broiler chicks in each group were slaughtered at 7 weeks of age, and their breast and thigh meat were collected. Lightness ($L^*$) and yellowness ($b^*$) in the T2 and T3 groups were higher than in the control group (p<0.05). The loss of breast meat during cooking was increased in the T1, T2, and T3 groups, and the water-holding capacity of the breast meat was also decreased in these three groups (p<0.05). The water-soluble protein solubility was lower in the T1, T2, and T3 groups than in the control group (p<0.05), but the salt-soluble protein solubility in these three groups was higher than that in the control group (p<0.05). The crude fat content of the breast meat was decreased and that of the thigh meat was increased as the level of FVM was increased (p<0.05). The unsaturated fatty acid content of the breast meat was higher in the T2 and T3 groups than in the control group (p<0.05). The free amino acid content was high in the T1, T2, and T3 groups compared to the control. In conclusion, these results showed that feeding with 3% and 5% FVM increased the L and b values in thigh meat, improved the salt-soluble protein solubility, and increased the content of unsaturated fatty acids and free amino acids.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.20 no.4
• /
• pp.766-783
• /
• 2006

This study describes philosophical background regarding '虛' in oriental medicine in an effort to understand the relationships among some of the Eastern philosophy in accordance with differentiated meanings in several resources by analyzing examples of '虛' in Huang Di Nei Jing. The various usages of '虛' used in Huang Di Nei Jing are as follows: naming; condition of pulse; emptiness; '太虛' which was referred universal space by Chinese ; insufficiency, lack or scarcity, deficiency ; and the description of vitality, mental faculties. 外丹修煉(training by external substances) had the attitude do that withdraw the death by taking external materials. The meaning of '虛'in 外丹修煉 is similar to that in oriental medicine in terms of deficiency. That is, both 外丹修煉 and the oriental medicine consider that the death and disease are caused by the deficiency of something. However, there also exists difference between 外丹修煉 and the oriental medicine. 外丹修煉 supplements through withdrawal prohibition due to the characteristic of unchangeability and stern or immortal while oriental medicine provides concrete object of deficiency. 精(essence of life), 氣(ki, functional activity), and fe(vitality) not only have been considered as basic component of human body, but they also have been an important subject of health preservation for longevity with health in Taoism and oriental medicine. In oriental medicine, 精 and 氣 have been perceived as physical basis of human body and 神 as controller. 內丹修煉(Training of internal active substances) 掠nds to return to '虛', the early state of life through individual training, and attempts to withdraw death through continuous recurrence. The oriental medicine and 內丹修煉 held great value of 神 among health preservation of 精, 氣, and 神. They seek theoretical basis from philosophical Taoism. However, '虛' in Taoism is different from that in training by internal substances and oriental medicine: '虛' in philosophical Taoism has metaphysical concept which refers overcome of life and death, but '虛' in 內丹修煉 and oriental medicine have empirical concept. '太虛' is considered as formless space where it is emp Dut filled with 氣. It is conceptualized with the premise of the relevant adaptation of human body to natural environment theory referring that the interaction between the heaven and the earth makes changes; all creation is originated , and human is affected by the interaction of the heaven and the earth. Furthermore, in $\ulcorner$運氣七篇$\lrcorner$ (Seven chapters described about the five circuit phases and the six atmospheric influences), the expression that the earth is in the center of '太虛' and huge amount of 氣 supports it proves that $\ulcorner$運氣七篇$\lrcorner$ adapts '渾天設'(Chaotic universe thee). In Taoism, '虛' is the grounds where all creation is generated in the optimal condition of Tao. As regards the aspect of mentality, it is the condition in which one can free from the dualistic concepts such as right and wrong, beauty and ugliness, life and death, and so on. Although the ultimate goal of oriental medicine, the achievement of longevity without sickness, might contrast with the Taoist belief that perceives life and death as the natural phenomena or the flowing of the 氣, and eliminates all international, the idea of Taoism that one should live substantial life with naivety, and make Harmony with the nature might be influential to the oriental medicine.

• (The)Study of the Eastern Classic
• /
• no.59
• /
• pp.405-428
• /
• 2015

Painting theorists in Song dynasty often spoke 'Li' when they talked about paintings. But 'Li' of the song dynasty is not limited to the 'ethics'. First, it includes visible 'natural's truth', such as the differences of 'geography' and 'physics' between the depicted objects. Second, it also includes 'common sense' which was based on both the observation and the thinking. The theorists thought if the 'common sense' was improper in the painting, the whole work was invalid. Thirdly, it also includes 'the reasonable sense in special situation', which requires great imagination ability and elaborative faculty. For example, when playing wind instruments and stringed instruments in the same time at the same concert, the painter should accurately draw the different gestures of musicians according to that the wind instrument is 'sound when the finger lift' and the stringed instrument is 'sound after the finger have left' in that moment. Fourthly, it includes 'art reason', theorists call it as 'ShenLi' or 'MiaoLi'. 'ShenLi' or 'MiaoLi' require the creator to join the spiritual concept besides his observation and thinking. For example, 'banana in snow' is neither observed available nor thought of available, but is the result of spiritual concept of creators for seeking everlasting. And at last, it certainly includes 'the principle of ethics'. Painters often highlighted the ethical relations of the feudal nation and the value of individual gentleman through the allegory story of figure painting and even the sudden composition in the landscape painting. 'Geography', 'physical', and 'common sense' are required the meticulous observation and the rational thinking for the painted object. And 'the reasonable sense in special situation', 'the art reason', 'the principle of ethics' are required enhancing painting style in the painting artistic conception and realm of life based on the nuanced observation, making 'technology' into the 'Tao'. This is the six reasonable increasing requirements for the painting work. Therefore, 'seeking the final reason' is the fundamental spirit of painting theory of Song Dynasty.

• Asian-Australasian Journal of Animal Sciences
• /
• v.32 no.7
• /
• pp.922-929
• /
• 2019

Objective: Mutations in low-density lipoprotein receptor (LDLR), which encodes a critical protein for cholesterol homeostasis and lipid metabolism in mammals, are involved in cardiometabolic diseases, such as familial hypercholesterolemia in pigs. Whereas microRNAs (miRNAs) can control LDLR regulation, their involvement in circulating cholesterol and lipid levels with respect to cardiometabolic diseases in pigs is unclear. We aimed to identify and analyze LDLR as a potential target gene of SSC-miR-20a. Methods: Bioinformatic analysis predicted that porcine LDLR is a target of SSC-miR-20a. Wild-type and mutant LDLR 3'-untranslated region (UTR) fragments were generated by polymerase chain reaction (PCR) and cloned into the pGL3-Control vector to construct pGL3 Control LDLR wild-3'-UTR and pGL3 Control LDLR mutant-3'-UTR recombinant plasmids, respectively. An miR-20a expression plasmid was constructed by inserting the porcine premiR-20a-coding sequence between the HindIII and BamHI sites in pMR-mCherry, and constructs were confirmed by sequencing. HEK293T cells were co-transfected with the miR-20a expression or pMR-mCherry control plasmids and constructs harboring the corresponding 3'-UTR, and relative luciferase activity was determined. The relative expression levels of miR-20a and LDLR mRNA and their correlation in terms of expression levels in porcine liver tissue were analyzed using reverse-transcription quantitative PCR. Results: Gel electrophoresis and sequencing showed that target gene fragments were successfully cloned, and the three recombinant vectors were successfully constructed. Compared to pMR-mCherry, the miR-20a expression vector significantly inhibited wild-type LDLR3'-UTR-driven (p<0.01), but not mutant LDLR-3'-UTR-driven (p>0.05), luciferase reporter activity. Further, miR-20a and LDLR were expressed at relatively high levels in porcine liver tissues. Pearson correlation analysis revealed that porcine liver miR-20a and LDLR levels were significantly negatively correlated (r = -0.656, p<0.05). Conclusion: LDLR is a potential target of miR-20a, which might directly bind the LDLR 3'-UTR to post-transcriptionally inhibit expression. These results have implications in understanding the pathogenesis and progression of porcine cardiovascular diseases.