• Microbiology and Biotechnology Letters
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• v.49 no.3
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• pp.283-288
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• 2021

This study investigated the anticancer activity of methanol extract from Platycarya strobilacea leaf in AGS human gastric cancer cells. We determined the cell viability effect of P. strobilacea using MTS assay. Apoptosis induction and cell cycle arrest were confirmed by fluorescein isothiocyanate and propidium iodide staining using cellometer K2. The mRNA expression levels of the Bcl-2 family were confirmed by reverse transcription-polymerase chain reaction. The cell viability was decreased in a dose-dependent manner treated with different concentrations of P. strobilacea. Total, early, and late apoptotic cells were dramatically increased, and the cell cycle was arrested at the sub-G1 phase. The mRNA expressions of Bcl-2 and Bcl-xL were reduced, whereas pro-apoptotic factors, Bax and Bak, were increased in a dose-dependent manner. These results suggested that P. strobilacea leaf extract induced significant apoptotic activity through an intrinsic mitochondria pathway.

• Journal of Plant Biotechnology
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• v.48 no.4
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• pp.246-254
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• 2021

Environmental stresses are estimated to have reduced global crop yields of wheat by 5.5%. However, traditional approaches for the transfer of resistance to these stresses in wheat plants have yielded limited results. In this regard, genetic transformation has undoubtedly opened up new avenues to overcome crop losses due to various abiotic stresses. Particle bombardment has been successfully employed for obtaining transgenic wheat. However, most of these procedures employ immature embryos, which are not available throughout the year. Therefore, the present investigation utilized mature seeds as the starting material and used the calli raised from three Moroccan durum wheat varieties as the target tissue for genetic transformation by the biolistic approach. The pANIC-5E plasmid containing the SINA gene for drought and salinity tolerance was used for genetic transformation. To enhance the regeneration capacity and transformation efficiency of the tested genotypes, the study compared the effect of copper supplementation in the induction medium (up to 5 μM) with the standard MS medium. The results show that the genotypes displayed different sensitivities to CuSO₄, indicating that the transformation efficiency was highly genotype-dependent. The integration of transgenes in the T₀ transformants was demonstrated by polymerase chain reaction (PCR) analysis of the obtained resistant plantlets with primers specific to the SINA gene. Among the three genotypes studied, 'Isly' showed the highest efficiency of 9.75%, followed by 'Amria' with 1.25% and 'Chaoui' with 1%.

• Journal of Korean Medicine Rehabilitation
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• v.29 no.2
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• pp.135-147
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• 2019

Objectives This study was performed to evaluate the effect of Pyrola japonica extract (NJ) and its principal constituent, homoarbutin (HA) on osteoclast differentiation and gene expression and bone resorption. The osteoclastogenesis and gene expression were determined in receptor activator of nuclear factor kappa B ligand (RANKL)-stimulated RAW264.7 cell. Methods In order to evaluate the effect of HA extracted from NJ on bone resorption, osteoclasts were used to be differentiated and formed by stimulating RAW264.7 cells with RANKL. Tartarate-resistant acid phosphatase (TRAP) (+) polynuclear osteoclast formation ability was evaluated, and differentiation control genes including cathepsin K, matrix metalloproteinases-9 (MMP-9), and TRAP in osteoclast differentiation were analyzed by real-time polymerase chain reaction (PCR). Immunoblotting was performed to measure the effect of mitogen-activated protein kinase (MAPK) factors on bone resorption, and the effect of osteoclasts on osteoclast differentiation was measured. Results Both NJ and high concentration of HA blocked RANKL-stimulated differentiation from RAW264.7 cell to TRAP-positive multinucleated cells. NJ reduced RANKL-induced expression of TRAP, cathepsin K. Both NJ and high concentration of HA inhibited RANKL-mediated expression of MMP-9, nuclear factor of activated T-cells, cytoplasmic 1, and cellular Jun-fos. NJ suppressed RANKL-stimulated expression of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase, tumor necrosis factor-alpha, and levels of interleukins. Both NJ and HA decreased bone resorption in osteoclast-induced bone pit formation model. Conclusions These results suggest that NJ and HA blocked bone resorption by decreasing RANKL-mediated osteoclastogenesis through down-regulation of genes for osteoclast differentiation.

• Korean Journal of Agricultural Science
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• v.46 no.3
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• pp.653-660
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• 2019

Ganoderma lucidum, an oriental polypore fungus and medicinal mushroom, has a long history of use for promoting health and longevity in Korea, China, and other Asian countries. This study was aimed at determining the anti-inflammatory activity and mechanism of action of Ganoderma lucidum in murine macrophage RAW 264.7 cells. Ganoderma lucidum was extracted with ethanol and freeze-dried. The anti-inflammatory effect (nitrite production) of Ganoderma lucidum extracts was tested using a nitric oxide (NO) colorimetric assay. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was performed to quantify the mRNA expression of cytokines including tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$), interleukin $(IL)-1{\beta}$, and IL-6. Western blotting was performed to measure the expression levels of inflammation-related proteins, such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor kappa B ($NF-{\kappa}B$) p65, and phosphorylated $NF-{\kappa}B$ p65. The NO colorimetric assay showed that NO production increased with the treatment of lipopolysaccharide in (LPS)-activated RAW 264.7 macrophages and decreased with the cotreatment of Ganoderma lucidum extracts and LPS. Ganoderma lucidum extracts repressed the mRNA expressions of cytokines, which were increased after the LPS treatment. In addition, Ganoderma lucidum extracts inhibited the LPS-induced expression of iNOS and COX-2 and the LPS-induced phosphorylation of $NF-{\kappa}B$ p65. These results suggest that the Ganoderma lucidum extracts exert an anti-inflammatory activity by inhibiting $NF-{\kappa}B$ related proteins and cytokines.

• Journal of Physiology & Pathology in Korean Medicine
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• v.33 no.2
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• pp.109-115
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• 2019

Aster Yomena (AY) has been used in traditional medicine to treat diseases such as obesity, hyperlipidemia, atherosclerosis, diabetes and osteoarthritis. However, protective effect of AY on acute pancreatitis (AP) has not been reported. The present study examined the anti-inflammatory effects of an ethanol extract of AY on cerulein-induced AP. AP was induced in mice by intraperitoneally injecting cerulein ($50{\mu}g/kg$) hourly for 6 times. 70% ethanol extract of AY (0.1, 0.2, and 0.5 g/kg) was orally administered for 1 week before acute pancreatitis induction. The mouse was killed at 6 hours after the final cerulein injection. The pancreas and lung were rapidly removed for histological examination and myeloperoxidase (MPO) assay. Blood samples were taken to determine serum amylase and lipase activity. In addition real-time reverse transcription-polymerase chain reaction (RT-PCR) was also performed to investigate mRNA expression of proinflammatory cytokines such as $TNF-{\alpha}$. $IL-1{\beta}$, and IL-6. Administration of AY significantly ameliorated pancreatic weight to body weight ratio, histological damages and MPO activity during AP. In addition, AY inhibited the serum amylase and lipase activity during AP. Also, mRNA expression of $TNF-{\alpha}$, $IL-1{\beta}$ and IL-6 were inhibited by AY against AP. Our results revealed that pre-treatment of AY reduces the severity of cerulein-induced AP. Therefore, AY may have a protective effect drug against AP.

• Entomological Research
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• v.48 no.5
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• pp.439-447
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• 2018

Adult mosquito surveillance was conducted from 2013 through 2014 at four cattle sheds, a wild bird refuge, and two residential areas located in Gyeongnam Province in the Republic of Korea. Adult mosquitoes were collected in black light traps from April 1, through November 30. Mosquito surveillance was conducted to figure out population densities of vector mosquitoes, possibly invaded mosquitoes and identify various virus infections at the selected sites. A total of 107,466 females comprising 14 species and 7 genera were collected from 2013 to 2014. The most common species collected were Culex tritaeniorhynchus Giles (63.8%), Anopheles sinensis s.l. (18.9%), Aedes vexans nipponii (Theobald) (7.7%), and Culex pipiens Coquillett (5.1%). Trap indices (TIs) varied widely for species over their range, due to geographical distribution and degree of association with rural and urban communities. The most collected An. sinensiss.l. and Cx. tritaeniorhynchus appeared at a cow shed in Hapcheon (TI 347.5) and a pigsty in Daejeo-1-dong, Busan (TI 1,040.8), respectively, due in part to their situation near breeding sites such as rice paddies. The bi-weekly population densities for mosquito species were variable for each of the years, apparently as a result of variable annual weather conditions. None of the mosquito species collected tested for the flavivirus including Japanese Encephalitis Virus, West Nile Virus, Dengue Virus, and Zika Virus infections by polymerase chain reaction (PCR) assay were positive.

• Journal of Ginseng Research
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• v.43 no.2
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• pp.261-271
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• 2019

Background: Protopanaxatriol (PPT) is an aglycone of ginsenosides, which has high medicinal values. Production of PPT from natural ginseng plants requires artificial deglycosylation procedures of ginsenosides via enzymatic or physicochemical treatments. Metabolic engineering could be an efficient technology for production of ginsenoside sapogenin. For PPT biosynthesis in Panax ginseng, damarenediol-II synthase (PgDDS) and two cytochrome P450 enzymes (CYP716A47 and CYP716A53v2) are essentially required. Methods: Transgenic tobacco co-overexpressing P. ginseng PgDDS, CYP716A47, and CYP716A53v2 was constructed via Agrobacterium-mediated transformation. Results: Expression of the three introduced genes in transgenic tobacco lines was confirmed by Reverse transcription-polymerase chain reaction (RT-PCR). Analysis of liquid chromatography showed three new peaks, dammarenediol-II (DD), protopanaxadiol (PPD), and PPT, in leaves of transgenic tobacco. Transgenic tobacco (line 6) contained $2.8{\mu}g/g$ dry weight (DW), $7.3{\mu}g/g$ DW, and $11.6{\mu}g/g$ DW of PPT, PPD, and DD in leaves, respectively. Production of PPT was achieved via cell suspension culture and was highly affected by auxin treatment. The content of PPT in cell suspension was increased 37.25-fold compared with that of leaves of the transgenic tobacco. Transgenic tobacco was not able to set seeds because of microspore degeneration in anthers. Transmission electron microscopy analysis revealed that cells of phloem tissue situated in the center of the anther showed an abnormally condensed nuclei and degenerated mitochondria. Conclusion: We successfully achieved the production of PPT in transgenic tobacco. The possible factors deriving male sterility in transgenic tobacco are discussed.

• The Journal of Internal Korean Medicine
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• v.40 no.3
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• pp.356-368
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• 2019

Objectives: This study was designed to evaluate the efficacy of Pharbitidis Semen (PS) on obesity using 3T3-L1 cells and high-fat-diet rats. Methods: After the administration of PS at a concentration of 0.125, 0.25, and 0.50 mg/ml in 3T3-L1 cells, the degree of differentiation of adipocytes was measured by Oil Red O staining. The experimental animals were divided into five groups in vivo. Normal group: diet-fed normal group (Normal), Control group (Control): high-fat-diet (HFD)-fed, Positive control group (PC): HFD+Contrave 200 mg/kg, PS_Low group: HFD+PS 250 mg/kg group, PS_High group: HFD+PS 500 mg/kg. Results: The total cholesterol, low density lipoprotein-cholesterol (LDL-cholesterol), and triglycerides (TG) content of the PS medication groups were significantly lower than those of the control group. In addition, high density lipoprotein cholesterol (HDL-cholesterol) tended to increase, and free fatty acid, total lipid, and phospholipid tended to decrease, but the decrease was not statistically significant. The administration of PS inhibited the differentiation of 3T3-L1 adipocytes, and a statistically significant difference was found in the group administered with 0.125 mg/ml. The mRNA expression of $PPAR-{\gamma}$ and leptin, which are involved in the differentiation of adipocytes, was observed using reverse transcription polymerase chain reaction (RT-PCR). The expressions of $PPAR-{\gamma}$ and leptin significantly decreased in comparison with the control and the positive control.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.32 no.3
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• pp.13-22
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• 2019

Objectives : Armeniacae semen is the seed of Prunus armenica L. var. ansu MAXIM, and this is classified into Rosaceae. Armeniacae semen has been used for centuries in traditional oriental medicine for the treatment of pain and inflammatory diseases. Amygdalin is the major compound of Armeniacae semen, and it is now being used for the treatment of pain and cancer. Methods : In the present study, we compared the effects of an aqueous extract of Armeniacae semen and a solution of amygdalin extracted from Armeniacae semen on lipopolysaccharide(LPS)-stimulated prostaglandin E2 synthesis and nitric oxide production in mouse BV-2 microglial cells. For this study, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay, reverse transcription-polymerase chain reaction(RT-PCR), prostaglandin E2 immunoassay and nitric oxide detection were performed on mouse BV-2 microglial cells. Results : In the present study, an aqueous extract of Armeniacae semen and an amygdalin solution extracted from Armeniacae semen suppressed prostaglandin E2 synthesis and nitric oxide production by inhibiting the LPS-induced enhancement of cyclooxygenase-2(COX-2) mRNA and the inducible nitric oxide synthase mRNA in mouse BV-2 cells. For the cyclooxygenase-1(COX-1) expression, an aqueous extract of Armeniacae semen showed a more potent suppression effect compared to the amygdalin solution. However, the amygdalin solution more potently suppressed the LPS-induced COX-2 mRNA expression compared to the aqueous extract of Armeniacae semen. Conclusions : As a result, aqueous extract of Armeniacae semen and amygdalin exert anti-inflammatory and analgesic effects.

• Journal of Plant Biotechnology
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• v.46 no.1
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• pp.1-8
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• 2019

The main goal of this study was to determine the genetic diversity among 36 grape cultivars grown in Palestine by using ISSR-polymerase chain reaction (PCR) fingerprints. Among the tested primers, 17 produced reasonable amplification products with high intensity and pattern stability. A total of 57 DNA fragments (loci) separated by electrophoresis on agarose gels were detected and they ranged in size, from 150 to 900 bp. Out of these fragments, 55 (88%) were polymorphic and 2 (3.5%) monomorphic. Our results also revealed an average of 3.1 loci per primer. A minimum of 1 and maximum of 10 DNA fragments were obtained (S-17, #820 and #841) and (S-31) primers, respectively. Therefore, the later primer (S-31) is considered to be the most powerful primer among the tested ones. The genetic distance matrix showed an average distance range of between 0.05 and 0.76. The maximum genetic distance value of 0.76 (24% similarity) was exhibited between the (Shami and Marawi.Hamadani.Adi) as well as (Bairuti and Marawi.Hamadani.Adi) genotypes. On the other hand, the lowest genetic distance of 0.05 (95% similarity) was exhibited between (Jandali.Tawel.Mofarad and Jandali. Kurawi.Mlzlz) along with (Shami.Aswad and Shami.mtartash. mlwn) genotypes. Furthermore, the UPGMA dendrogram generally clusters the grape cultivars into eight major clusters in addition to an isolated genotype. Based on these figures, the cultivars tested in this study could be characterized by large divergence at the DNA level. This is taking the assumption that our region has a very rich and varied clonal grape genetic structure.