• Asian Pacific Journal of Cancer Prevention
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• v.14 no.2
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• pp.685-689
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• 2013

Objective: To study the effects of down-regulation of HDAC6 expression on proliferation, cell cycling and migration of esophageal squamous cell carcinoma (ESCC) cells and related molecular mechanisms. Methods: ESCC cell line EC9706 cells were randomly divided into untreated (with no transfection), control siRNA (transfected with control siRNA) and HDAC6 siRNA (transfected with HDAC6 small interfering RNA) groups. Effects of HDAC6 siRNA interference on expression of HDAC6 mRNA and protein in EC9706 cells were investigated by semi-quantitative RT-PCR, Western blotting and immunocytochemistry methods. Effects of down-regulation of HDAC6 expression on cell proliferation, cell cycle, and cell migration were studied using a CCK-8 kit, flow cytometry and Boyden chambers, respectively. Changes of mRNA and protein expression levels of cell cycle related factor (p21) and cell migration related factor (E-cadherin) were investigated by semi-quantitative RT-PCR and Western blotting methods. Results: After transfection of HDAC6 siRNA, the expression of HDAC6 mRNA and protein in EC9706 cells was significantly downregulated. In the HDAC6 siRNA group, cell proliferation was markedly inhibited, the percentage of cells in G0/G1 phase evidently increased and the percentage of cells in S phase decreased, and the number of migrating cells significantly and obviously decreased. The mRNA and protein expression levels of p21 and E-cadherin in the HDAC6 siRNA group were significantly higher than those in the untreated group and the control siRNA group, respectively. Conclusions: HDAC6 siRNA can effectively downregulate the expression of HDAC6 mRNA and protein in EC9706 cells. Down-regulation of HDAC6 expression can obviously inhibit cell proliferation, arrest cell cycling in the G0/G1 phase and reduce cell migration. The latter two functions may be closely related with the elevation of mRNA and protein expression of p21 and E-cadherin.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.9
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• pp.5441-5447
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• 2013

Purpose: To demonstrate the value of sequential determinations of pelvic drainage in the identification of increased risk of anastomotic leakage (AL) after anterior resection for rectal cancer with a double stapling technique. Patients and Methods: Between January 2004 and December 2011, data for the daily postoperative pH of pelvic drainage fluid in 753 consecutive patients with rectal cancer who initially underwent anterior resection with a double stapling technique were reviewed. All patients experienced a total mesorectal excision. Patients with anastomotic leakage (Group AL, n=57) were compared to patients without leakage (Group nAL, n=696). Patients with perioperatively abdominopelvic implants that were likely to affect pH value (determined at $25^{\circ}C$) other than leakage were excluded. Mean postoperative values were compared. Results: Anastomotic leakage was noted in 57 (7.6%) of 753 patients with rectal cancer. The diagnosis of AL was made between the $6^{th}$ and $12^{th}$ postoperative day (POD; mean $8^{th}$ POD). There was no significance of the daily average values of pH on POD1 & 2 in group AL while a significantly sharp declining mean pH value reached its diagnostic point of AL (p<0.001) on POD3. A cut-off value of 6.978 on the $3^{rd}$ POD maximized the sensitivity (98.7.0%) and specificity (94.7%) in assessing the risk of leakage. Conclusion: According to these results, an early and persistent declining of pH value of pelvic drainage fluid after rectal surgery with anastomosis, is a marker of AL. A cut-off value of 6.798 determined at $25^{\circ}C$ on POD3 maximizes sensitivity and specificity.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.9
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• pp.5527-5531
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• 2013

Background and Aim: Increasing evidence correlates the presence of systemic inflammation with poor survival in patients with hepatocellular carcinoma (HCC). The aim of this study was to investigate the prognostic significance of the blood neutrophil-to-lymphocyte ratio (NLR) in patients with advanced HCC who received sorafenib monotherapy. Methods: A total of sixty-five patients with advanced HCC, not eligible for locoregional therapy, treated with sorafenib were enrolled. Potential prognostic factors such as age, gender, tumoral characteristics, performance status and NLR were analyzed. Results: Median OS and TTP for the entire cohort were 10.0 months (95%CI, 7.6-12.3 months) and 4.5 months (95% CI, 4.0-4.9 months). The mean NLR at baseline was 2.89. The median OS of patients with a high NLR (>4) was 6.5 months (95%CI, 5.2-7.7 months) compared with 12.5 months (95%CI, 9.9-15.0) for patients with a normal NLR (${\leq}4$) (P=0.01). Age ${\leq}65$, NLR>4, extrahepatic metastases and vascular invasion were all predictors of poorer overall survival. Multivariate analysis showed that NLR > 4, vascular invasion and extrahepatic metastases were independent predictors of poorer overall survival. The median TTP of patients with a high NLR was 2.5 months (95%CI, 1.4-3.6 months) compared with 4.5 months (95%CI, 3.9-5.1 months) for patients with a normal NLR (P=0.012). Conclusions: High baseline NLR was associated with worse OS and TTP for patients with advanced HCC treated with sorafenib.

• Journal of Korean Medical classics
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• v.29 no.2
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• pp.135-150
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• 2016

Objectives : KangHaiChengZhiLun (亢害承制論; If Excess Brings Harm, Lifing Qi (承氣) Restrains) was originally a theory that explained how the realms of nature remain in harmony and equilibrium. It later became an important theory for clinical trials of Traditional Chinese Medicine, explaining the physiological and pathological mechanism. Methods : The researcher considered all the annotations and the original text of SuWen(素問), LiuWeiZhiDaLun(六微旨大論) and theories of medical practitioners who applied KangHaiChengZhiLun(亢害承制論) to their clinical trials. Results & Conclusions : Wangbing (王氷) went with a theory that phenomena of Lifting Qi (承氣) take place in the realms of nature when Qi (氣) flourishes. In XinJiaoZheng(新校正), he wrote about two theories: one was that Six Kinds of Natural Factors (六氣) first work as the main Qi (本氣) but later bring about Lifting Qi. (終見下承之氣說); the other was that excessive Stagnation Qi (鬱氣) can be exploded and invite another accompanying Qi, Lifting Qi. (甚者兼其下承之氣說) Liuwansu (劉完素) had a theory that if Six Kinds of Natural Factors go disproportionately excessive, it becomes accompanied by imaginary Qi (假象) that conquers self. (反兼勝己之化說) $Wangl{\ddot{u}}$(王履) maintained that Lifting Qi usually works as a means to prevent Six Kinds of Natural Factors (六氣) from becoming rampant; but when Six Kinds of Natural Factors become overly excessive, Lifting Qi restrains them in order to maintain equilibrium. (防之與克勝說) Yutuan explained that since Excessive Qi (亢氣) does damage to the mother of Lifting Qi, Lifting Qi restrains Excessive Qi to protect Original Qi (元氣), its mother. (護救承者之元氣說) Gongtingxian was in favor of two theories: one argued that causes and symptoms of a disease differ from each other. (體用不同說); the other said that diseases are naturally cured if the patient finds out the time when Lifting Qi gains strength. (得承之時自愈說) Mashi (馬蒔) had a theory that Lifting Qi is generated when Six Kinds of Natural Factors are prosperous and reveals itself when its season comes. (極則生承氣 至本位著說) Zhangjiebin (張介賓) asserted that when Six Kinds of Natural Factors are thriving, Lifting Qi, as a restraining force, is generated to disperse the thriving natural factors and leads to a new one. (前之退而後之進說) Zhangqi (張琦)'s argument was that if Lifting Qi restrains the main Qi, a son of the main Qi is generated and every four season goes in harmony. (承氣制則生化說) Hemengyao (何夢瑤) had an argument that a son of the restrained Qi succeeds to its father and later achieves equilibrium by restraining Excessive Qi. (被克承父 制之平衡說).

• Korean Journal of Medicinal Crop Science
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• v.7 no.4
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• pp.245-250
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• 1999

Callus growth of fusaric aicd-tolerance cell lines was different depending on fusaric acid concentrations. But callus growth on medium with fusaric acid was higher than that on medium without fusaric acid. Especially, RF-9, RF-11 and RF-15 showed high callus growth at $100\;{\mu}M$ fusaric acid. After subculturing on medium without fusaric acid for 5 weeks, fusaric acid -tolerant stability was investigated. Cell lines at $10{\mu}M$ fusaric acid were showed over 60% callus growth, callus growth rate at $100{\mu}M$ fusaric acid was decreased until 30-80% of control. Regeneration capacity of fusaric acid-tolerant cell lines was different depending on fusaric acid concentrations. Thirteen cell lines regenerated the shoot over at $50{\mu}M$ fusaric acid, and only two cell lines were not regenerated.

• Reproductive and Developmental Biology
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• v.32 no.3
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• pp.167-173
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• 2008

Somatic cells such as oviduct epithelial cell, uterine epithelial cell, cumulus-granulosa cell and buffalo rat river cell has been used to establish an effective culture system for bovine embryos produced in in vitro. But nitric oxide (NO) metabolites secreted from somatic cells were largely arrested the development of bovine in vitro matured/ in vitro fertilized (IVM/IVF) embryos, suggesting that NO was induced the embryonic toxic substance into culture medium. The objective of this study was to investigate whether BOEC co-culture system can ameliorate the NO-mediated oxidative stress in the culture of bovine IVM/IVF embryos. Therefore, we evaluated the developmental rate of bovine IVM/IVF embryos under BOEC co-culture system in the presence or absence of sodium nitroprusside (SNP), as a NO donor, and also detected the expression of growth factor (TGF-$\beta$, EGF and IGFBP) and apoptosis (Caspase-3, Bax and Bcl-2) genes. The supplement of SNP over 5 uM was strongly inhibited blastocyst development of bovine IVM/IVF embryos than in control and 1 uM SNP group (Table 2). The developmental rates beyond morulae stages of bovine IVM/IVF embryos co-cultured with BOEC regardless of SNP supplement (40.4% in 5 uM SNP+ BOEC group and 65.1% in BOEC group) were significantly increased than those of control (35.0%) and SNP single treatment group (23.3%, p<0.05: Table 3). The transcripts of Bax and Caspase-3 genes were detected in all experiment groups (1:Isolated fresh cell (IFC), 2:Primary culture cell (PCC), 3:PCC after using the embryo culture, 4: PCC containing 5 uM SNP and 5: PCC containing 5 uM SNP after using the embryo culture), but Bcl-2 gene was not detected in IFC and PCC (Fig. 1). In the expression of growth factor genes, TGF-$\beta$ gene was found in all experimental groups, and EGF and IGFBP genes were not found in IFC and PCC (Fig. 2). These results indicate that BOEC co-culture system can increase the development beyond morula stages of bovine IVM/IVF embryos, possibly suggesting the alleviation of embryonic toxic substance like nitric oxide.

• Parasites, Hosts and Diseases
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• v.52 no.1
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• pp.21-26
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• 2014

Dirofilaria immitis (heartworm) infections affect domestic dogs, cats, and various wild mammals with increasing incidence in temperate and tropical areas. More sensitive antibody detection methodologies are required to diagnose asymptomatic dirofilariasis with low worm burdens. Applying current transcriptomic technologies would be useful to discover potential diagnostic markers for D. immitis infection. A filarial homologue of the mammalian translationally controlled tumor protein (TCTP) was initially identified by screening the assembled transcriptome of D. immitis (DiTCTP). A BLAST analysis suggested that the DiTCTP gene shared the highest similarity with TCTP from Loa loa at protein level (97%). A histidine-tagged recombinant DiTCTP protein (rDiTCTP) of 40 kDa expressed in Escherichia coli BL21 (DE3) showed immunoreactivity with serum from a dog experimentally infected with heartworms. Localization studies illustrated the ubiquitous presence of rDiTCTP protein in the lateral hypodermal chords, dorsal hypodermal chord, muscle, intestine, and uterus in female adult worms. Further studies on D. immitis-derived TCTP are warranted to assess whether this filarial protein could be used for a diagnostic purpose.

• Plant Biotechnology Reports
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• v.4 no.4
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• pp.253-260
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• 2010

Plant secondary metabolites have always been a focus of study due to their important roles in human medicine and nutrition. We transferred the isoflavone synthase (IFS) gene into soybean [Glycine max (L.) Merr.] using the Agrobacterium-mediated transformation method in an attempt to produce transformed soybean plants which produced increased levels of the secondary metabolite, isoflavone. Although the trial to produce transgenic plant failed due to unestablished hygromycin selection, transformed callus cell lines were obtained. The induction rate and degree of callus were similar among the three cultivars tested, but light illumination positively influenced the frequency of callus formation, resulting in a callus induction rate of 74% for Kwangan, 67% for Sojin, and 73% for Duyou. Following seven to eight subcultures on selection media, the isoflavone content of the transformed callus lines were analyzed by high-performance liquid chromatography. The total amount of isoflavone in the transformed callus cell lines was three- to sixfold higher than that in control callus or seeds. Given the many positive effects of isoflavone on human health, it may be possible to adapt our transformed callus lines for industrialization through an alternative cell culture system to produce high concentrations of isoflavones.

• Korean Journal of Food Science and Technology
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• v.42 no.4
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• pp.420-423
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• 2010

The proanthocyanidin fraction was isolated from the wild grape (Vitis amurensis) peel and its antioxidant capacities were examined to promote the utilization of wild grape by-products. The 70% acetone crude extract of the wild grape peel was fractionated with hexane, ethyl acetate, and water. The ethyl acetate fraction was applied to a Sephadex LH-20 column chromatograph, which was eluted with 50% methanol, 75% methanol, and 75% acetone. The proanthocyanidin characteristics and contents of the isolated fractions were investigated by the vanillin-$H_2SO_4$ and BuOH-HCl methods. Fraction 6 had the highest proanthocyanidin content ($49.35{\pm}2.75\;g%$) among the isolated fractions. The antioxidant activities of the proanthocyanidin fraction were examined by DPPH radical scavenging, FRAP assay, and total phenolic contents. The FRAP values and total phenolic contents of the fractions ranged from 3.54 to 32.25 mmol/kg and from 4.48 to 50.80 g/100 g, respectively. The proanthocyanidin contents was strongly correlated with DPPH radical scavenging activities, FRAP values, and total phenolic contents.

• Molecules and Cells
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• v.39 no.2
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• pp.103-110
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• 2016

As a degenerative joint disease, osteoarthritis (OA) constitutes a major cause of disability that seriously affects the quality of life of a large population of people worldwide. However, effective treatment that can successfully reverse OA progression is lacking until now. The present study aimed to determine whether two small non-coding RNAs miR-29a and miR-140, which are significantly down-regulated in OA, can be applied together as potential therapeutic targets for OA treatment. MiRNA synergy score was used to screen the miRNA pairs that potentially synergistically regulate OA. An in vitro model of OA was established by treating murine chondrocytes with IL-$1{\beta}$. Transfection of miR-29a and miR-140 via plasmids was investigated on chondrocyte proliferation and expression of nine genes such as ADAMTS4, ADAMTS5, ACAN, COL2A1, COL10A1, MMP1, MMP3, MMP13 and TIMP metallopeptidase inhibitor 1 (TIMP1). Western blotting was used to determine the protein expression level of MMP13 and TIMP1, and ELISA was used to detect the content of type II collagen. Combined use of miR-29a and miR-140 successfully reversed the destructive effect of IL-$1{\beta}$ on chondrocyte proliferation, and notably affected the MMP13 and TIMP1 gene expression that regulates extracellular matrix. Although co-transfection of miR-29a and miR-140 did not show a synergistic effect on MMP13 protein expression and type II collagen release, but both of them can significantly suppress the protein abundance of MMP13 and restore the type II collagen release in IL-$1{\beta}$ treated chondrocytes. Compared with single miRNA transfection, cotransfection of both miRNAs exceedingly abrogated the suppressed the protein production of TIMP1 caused by IL-$1{\beta}$, thereby suggesting potent synergistic action. These results provided1novel insights into the important function of miRNAs' collaboration in OA pathological development. The reduced MMP13, and enhanced TIMP1 protein production and type II collagen release also implies that miR-29a and miR-140 combination treatment may be a possible treatment for OA.