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International Wound Journal logoLink to International Wound Journal
. 2015 Oct 28;13(Suppl 1):42–46. doi: 10.1111/iwj.12530

Effects of Panax ginseng extract on human dermal fibroblast proliferation and collagen synthesis

Geum‐Young Lee 1, Kang‐Gyun Park 1, Sik Namgoong 1, Seung‐Kyu Han 1,, Seong‐Ho Jeong 1, Eun‐Sang Dhong 1, Woo‐Kyung Kim 1
PMCID: PMC7949929  PMID: 26507878

Abstract

Current studies of Panax ginseng (or Korean ginseng) have demonstrated that it has various biological effects, including angiogenesis, immunostimulation, antimicrobial and anti‐inflammatory effects. Therefore, we hypothesised that P. ginseng may also play an important role in wound healing. However, few studies have been conducted on the wound‐healing effects of P. ginseng. Thus, the purpose of this in vitro pilot study was to determine the effects of P. ginseng on the activities of fibroblasts, which are key wound‐healing cells. Cultured human dermal fibroblasts were treated with one of six concentrations of P. ginseng: 0, 1, 10 and 100 ng/ml and 1 and 10 µg/ml. Cell proliferation was determined 3 days post‐treatment using the 3‐(4, 5‐dimethylthiazol‐2‐yl)‐2, 5‐diphenyl tetrazolium bromide assay, and collagen synthesis was evaluated by the collagen type I carboxy‐terminal propeptide method. Cell proliferation levels and collagen synthesis were compared among the groups. The 10 ng/ml to 1 µg/ml P. ginseng treatments significantly increased cell proliferation, and the 1 ng/ml to 1 µg/ml concentrations significantly increased collagen synthesis. The maximum effects for both parameters were observed at 10 ng/ml. P. ginseng stimulated human dermal fibroblast proliferation and collagen synthesis at an optimal concentration of 10 ng/ml.

Keywords: Fibroblast, Ginseng, Panax ginseng, Wound healing

Introduction

Ginseng has been used for a wide range of preventative and therapeutic purposes in traditional herbal medicine for over 4000 years. It has been used to increase energy, vitality and as a method to slow ageing. Among several types of ginseng, Panax ginseng is the most commonly used medicinal ginseng because its efficacy is higher than that of other ginsengs 1. The term Panax is derived from the Greek word for all‐healing. Korea has been the largest supplier of P. ginseng, which is the world's best quality ginseng of all types grown in Korea. Ginseng products were marketed in more than 35 countries in 2010, and half of the sales came from South Korea. Therefore, P. ginseng is commonly referred to as ‘Korean ginseng’ or ‘true ginseng’.

The pharmacological effects of P. ginseng have been discovered through extensive research. P. ginseng dilates blood vessels, controls cholesterol levels and has anti‐cancer, anti‐ageing and anti‐allergic properties 2, 3, 4, 5, 6. P. ginseng also benefits the cardiovascular and dermatological systems 7, 8, 9. In particular, current studies demonstrate that P. ginseng has anti‐inflammatory, antioxidant, antimicrobial and immunomodulating effects 10, 11, 12, 13, 14, 15, 16, 17, 18. In addition, P. ginseng improves protein synthesis, neovascularization and angiogenesis 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29. Considered together, these features suggest that P. ginseng may play an important role in the wound‐healing process. However, a direct role for P. ginseng in wound healing has not been demonstrated.

Wound healing involves complex physiological and biological mechanisms that can be classified into inflammatory, proliferative and remodelling phases 30. These processes result from interactions among the various cell types that respond to the wound environment by providing the desired growth factors and other substances to the wound site 31. Fibroblasts are the single most important cell type, not only during the proliferative phase but also throughout the remodelling phase. Therefore, we hypothesised that P. ginseng would be useful for wound healing if it stimulates fibroblast activities. For example, new dressing material impregnated with P. ginseng could be developed.

The ultimate goal of our P. ginseng project was to determine its direct effects on wound healing. The present pilot study was designed to examine these effects for the first time.

Materials and methods

Human dermal fibroblast culture

Cultured human dermal fibroblasts were obtained from cryopreserved cells derived from the dermis of healthy adults (n = 10). The cell donors provided informed consent for their cells to be used for research purposes.

The cells were cultured in Dulbecco's Modified Eagle Medium/Ham's F‐12 nutrient (DMEM/F‐12; Gibco; Grand Island, NY) containing 10% foetal bovine serum (FBS; Gibco; Grand Island, NY) and 25 µg/ml gentamycin at 37°C in a 5% CO2/95% air atmosphere at 100% humidity. The fibroblasts were dissociated with trypsin, diluted 2·7‐fold with Mg2+ and Ca2+‐free Dulbecco's phosphate‐buffered saline (DPBS; Gibco; Grand Island, NY) and collected by centrifugation at 450g for 17 minutes. The cells were washed twice in 40 ml DPBS and resuspended in 5 ml DPBS. Cell density was determined using a haemocytometer, and cell viability was assessed with the trypan blue dye exclusion assay. Two‐passage cell cultures were used in the experiments.

Panax ginseng treatment

Ginsenoside Rb1 (G‐Rb1), which is the representative component of P. ginseng, was used for the study. The G‐Rb1 reagent was purchased from SigmaAldrich Korea, Ltd (Seoul, Korea). G‐Rb1 was diluted with DPBS to make 0, 1, 10 and 100 ng/ml and 1 and 10 µg/ml treatment groups. Cultured fibroblasts were seeded into a 96‐well culture plate at 2 × 103 cells/well in DMEM/F‐12 containing 5% FBS. The wells were treated with the six G‐Rb1 concentrations. The cells were incubated for 72 hours at 37°C in 5% CO2 with 100% humidity.

Cell proliferation assay

Cell proliferation was determined by the 3‐(4, 5‐dimethylthiazol‐2‐yl)‐2, 5‐diphenyl tetrazolium bromide (MTT) assay. Next 10 µl of MTT was added to a 100 µl cell monolayer in each of the 96 wells, and the plate was incubated for 3 hours at 37°C. Absorbance was measured at a test wavelength of 450 nm using an enzyme‐linked immunosorbent assay reader with a reference wavelength of 600 nm to evaluate cell proliferation.

Collagen synthesis assay

A collagen type I carboxy‐terminal propeptide (CICP) enzyme immunoassay was performed using a Metra CICP kit (Quidel, San Diego, CA) according to the manufacturer's instructions. Then, 100 µl of diluted culture supernatant was added to each well of a monoclonal anti‐CICP antibody‐coated plate, and the plate was incubated at room temperature for 2 hours. A 100 µl aliquot of rabbit anti‐CICP antisera was added for 50 minutes followed by 100 µl of goat anti‐rabbit alkaline phosphatase conjugate for 50 minutes. Then, the p‐nitrophenyl phosphate (pNPP) substrate was added for 30 minutes to quantify CICP. The reaction was stopped, and collagen synthesis was measured at 405 nm.

Statistical analyses

Statistical analyses were performed using the Kruskal–Wallis analysis of variance. Pair‐wise comparisons were made for parameters with statistical significance using the Wilcoxon signed‐rank test after the Kruskal–Wallis analysis. Results are expressed as mean and standard deviation. Statistical significance was accepted at P‐values <0·05. The data were analysed using SPSS 17.0 software (SPSS, Inc., Chicago, IL).

Results

Cell proliferation

Cell proliferation in all G‐Rb1‐treated groups increased compared with that in the control group. Significant differences were detected in the 10 ng/ml to 1 µg/ml concentrations (P < 0·05). Maximum cell proliferation was observed at the 10 ng/ml G‐Rb1 concentration (Figure 1). The significant differences among the tested concentrations are shown in Table 1.

Figure 1.

IWJ-12530-FIG-0001-c

Cell proliferation results (*P < 0·05, Wilcoxon signed‐rank test).

Table 1.

Significant differences in cell proliferation

IWJ-12530-TBL-0001-c

Collagen synthesis

Collagen synthesis in all G‐Rb1‐treated groups increased when compared with that in the control group. Significant differences were detected at the 1 ng/ml to 1 µg/ml concentrations (P < 0·05). Maximum collagen synthesis was observed at the 10 ng/ml G‐Rb1 concentration (Figure 2). The significant differences among the tested concentrations are shown in Table 2.

Figure 2.

IWJ-12530-FIG-0002-c

Collagen synthesis results (*P < 0·05, Wilcoxon signed‐rank test).

Table 2.

Significant differences in collagen synthesis

IWJ-12530-TBL-0002-c

Discussion

Ginseng is comprised of saponins and non‐saponins. The main active ginseng constituent is saponin. Saponins are a class of chemical compounds found abundantly in various plant species. In particular, ginseng has high saponin content. Ginseng saponin has a different chemical structure than that of other plant saponins and has extensive biological activities. Therefore, this distinct saponin is called ginsenoside. Ginsenosides are found nearly exclusively in the Panax species (ginseng); thus, they are also called panaxosides. More than 150 naturally occurring ginsenosides have been isolated from the roots, leaves/stems, fruits and flower heads of ginseng. Ginsenosides have been the target of many studies as they are believed to be the main active components behind ginseng efficacy claims. Ginsenosides are often split into the Rb1 group (characterised by the presence of protopanaxadiols: Rb1, Rb2, Rc and Rd) and the Rg1 group (protopanaxatriols: Rg1, Re, Rf and Rg2). The remaining components are called non‐saponins, which include polysaccharides, polyacetylenes, peptides and amino acids. G‐Rb1 is a representative P. ginseng component with a variety of biological activities, including anti‐inflammatory, antioxidant and antimicrobial effects. G‐Rb1 also improves protein synthesis, neovascularization or angiogenesis and immunity.

Over the past few decades, many wound specialists have been studying exclusively the stimulation of wound healing. Optimum wound healing requires a well‐orchestrated integration of complex biological and molecular events, including cell migration, proliferation, extracellular matrix deposition and remodelling. Therefore, the activities of key cells are essential to mediate successful wound healing. These cells release various cytokines and regulate the wound‐healing process. In particular, fibroblasts are the single most important cell type as they have numerous functions, including production of collagen, growth factors, antioxidants and a balance of matrix‐producing proteins and protease enzymes. Fibroblasts also play an essential role initiating tissue remodelling during wound recovery. One of the main contributing factors to delayed or non‐healing of chronic wounds is decreased fibroblast activities 32. Therefore, this pilot study focused on the effect of G‐Rb1 on fibroblast activities.

The effects of G‐Rb1 on cell activities have been reported previously but are controversial, mainly because of the differences in the proliferative responses to G‐Rb1 in different cell lines. A previous in vitro study showed that G‐Rb1 has no effect on human keratinocyte (HaCaT) proliferation 27. However, another study demonstrated that G‐Rb1 increases the viability of human retinal pigment epithelial cells 22. An in vivo study showed that G‐Rb1 inhibits the chemoinvasion of endothelial cells during neovascularization 26. However, another study reported that G‐Rb1 increases the number of blood vessels in burn wound areas of mice 27. Schwann cell proliferation is significantly inhibited at 1 mg G‐Rb1/ml, whereas 10 µg/ml induces proliferation 33. The effect of G‐Rb1 on collagen synthesis is also controversial. One study showed that G‐Rb1 enhances collagen production in HaCaT cells 27; however, G‐Rb1 decreases collagen levels in normal rat renal tubular epithelial cells (NRK‐52E) 34. Similarly, the effects of G‐Rb1 on cell function have been mixed. Furthermore, the efficacy of G‐Rb1 on human dermal fibroblasts has not been validated. In this study, we showed that G‐Rb1 had significant positive effects on dermal fibroblast proliferation and collagen synthesis, which are essential factors during wound healing. Maximum fibroblast activities were observed at 10 ng/ml.

Our results could be clinically applicable to wound dressings. P. ginseng is already used in various fields such as the making of soaps and cosmetics and as a beverage flavouring. New wound dressings impregnated with P. ginseng could be another example. A wound dressing is an adjunct applied to a wound to promote healing. Various dressing materials have been commercialised with the aim of supplementing the shortfalls of conventional gauze dressings. Film, hydrocolloid, hydrogel, foam, hydrofibre, composite and biological dressings are available to suit the wound condition. Functional reagent‐based dressings are now commonly used to help wound healing. Ibuprofen, silver and povidone‐iodine‐impregnated dressings are examples. If P. ginseng has a positive effect on wound healing, it could be added to the dressing material to accelerate healing. In particular, P. ginseng may be more likely to facilitate healing in older patients with chronic wounds because they commonly have poor blood circulation, a weak immune system, deficient nutritional factors and decreased cell activities. Additionally, chronic wounds usually persist in the inflammatory stage for a long time. As mentioned earlier, a number of clinical and experimental studies have confirmed that P. ginseng has a variety of positive effects in humans, including angiogenesis, immunostimulation, and antimicrobial and anti‐inflammatory actions. Considering these effects and the results of the present study, P. ginseng may be a good treatment option for healing chronic wounds. However, the fibroblasts used in the present study were derived from the dermis of healthy adults. A further study will be necessary to confirm the effect of P. ginseng on the activity of fibroblasts obtained from chronic wound patients (e.g., diabetic fibroblasts) as fibroblasts in chronic wounds commonly have a lower proliferative potential and an attenuated production of extracellular matrix components.

P. ginseng generally is well tolerated. Although adverse effects of P. ginseng have been reported in cases where it was administered orally, including capsules, liquids or powders, they were mild and reversible 1, 35, 36. Shergis et al. conducted a systematic review to evaluate randomised controlled trials for P. ginseng in patients with any type of disease or in healthy individuals. Of the 40 studies where adverse events were reported, 16 studies had no reported adverse events and 24 studies had 135 minor events. Minor events included diarrhoea, sleeplessness, palpitations, headache, nausea and mild hepatic dysfunction. No severe adverse events were reported from any study. In addition, there have been no reports demonstrating side effects of P. ginseng administered topically.

There are limitations to the present study. Although cell proliferation and collagen synthesis are key factors in examining wound‐healing potential, other parameters such as growth factors and glycosaminoglycans are also important contributing factors. Further studies may be required to determine explicitly the effect of P. ginseng on the activities of fibroblasts. It was difficult to assess the effect of P. ginseng on the overall wound‐healing process as in vivo wound healing is a complex process requiring multiple factors. Future studies of how P. ginseng treatment assists in wound healing in vivo will be needed. Moreover, the clinical applicability of P. ginseng requires a detailed evaluation. In addition, this study focused only on the possibility of using P. ginseng to increase fibroblast activities in vitro. The mechanism of action should be further investigated, including histological or biochemical analyses, to confirm the effects of ginseng treatment.

Conclusion

P. ginseng stimulated human dermal fibroblast proliferation and collagen synthesis at an optimal concentration of 10 ng/ml.

References

  • 1. Kiefer D, Pantuso T. Panax ginseng. Am Fam Physician 2003;68:1539–42. [PubMed] [Google Scholar]
  • 2. Yang F, Zheng Y, Li D, Deng W. Effect of shenfu injection on microcirculation. Sheng Wu Yi Xue Gong Cheng Xue Za Zhi 2003;20:91–4. [PubMed] [Google Scholar]
  • 3. Yamamoto M, Uemura T, Nakama S, Uemiya M, Kumagai A. Serum HDL‐cholesterol‐increasing and fatty liver‐improving actions of Panax ginseng in high cholesterol diet‐fed rats with clinical effect on hyperlipidemia in man. Am J Chin Med 1983;11:96–101. [DOI] [PubMed] [Google Scholar]
  • 4. Xiaoguang C, Hongyan L, Xiaohong L, Zhaodi F, Yan L, Lihua T, Rui H. Cancer chemopreventive and therapeutic activities of red ginseng. J Ethnopharmacol 1998;60:71–8. [DOI] [PubMed] [Google Scholar]
  • 5. Metori K, Furutsu M, Takahashi S. The preventive effect of ginseng with du‐zhong leaf on protein metabolism in aging. Biol Pharm Bull 1997;20:237–42. [DOI] [PubMed] [Google Scholar]
  • 6. Park EK, Choo MK, Kim EJ, Han MJ, Kim DH. Antiallergic activity of ginsenoside Rh2. Biol Pharm Bull 2003;26:1581–4. [DOI] [PubMed] [Google Scholar]
  • 7. Kim H, Chen X, Gillis CN. Ginsenosides protect pulmonary vascular endothelium against free radical‐induced injury. Biochem Biophys Res Commun 1992;189:670–6. [DOI] [PubMed] [Google Scholar]
  • 8. Kanzaki T, Morisaki N, Shiina R, Saito Y. Role of transforming growth factor‐beta pathway in the mechanism of wound healing by saponin from Ginseng Radix rubra. Br J Pharmacol 1998;125:255–62. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9. Hinz B. Formation and function of the myofibroblast during tissue repair. J Invest Dermatol 2007;127:526–37. [DOI] [PubMed] [Google Scholar]
  • 10. Cabral de Oliveira AC, Perez AC, Merino G, Prieto JG, Alvarez AI. Protective effects of Panax ginseng on muscle injury and inflammation after eccentric exercise. Comp Biochem Physiol C Toxicol Pharmacol 2001;130:369–77. [DOI] [PubMed] [Google Scholar]
  • 11. Kim SH, Park KS, Chang MJ, Sung JH. Effects of Panax ginseng extract on exercise‐induced oxidative stress. J Sports Med Phys Fitness 2005;45:178–82. [PubMed] [Google Scholar]
  • 12. Ahn JY, Song JY, Yun YS, Jeong G, Choi IS. Protection of Staphylococcus aureus‐infected septic mice by suppression of early acute inflammation and enhanced antimicrobial activity by ginsan. FEMS Immunol Med Microbiol 2006;46:187–97. [DOI] [PubMed] [Google Scholar]
  • 13. Jie YH, Cammisuli S, Baggiolini M. Immunomodulatory effects of Panax ginseng C.A. Meyer in the mouse. Agents Actions 1984;15:386–91. [DOI] [PubMed] [Google Scholar]
  • 14. Hong CE, Lyu SY. Anti‐inflammatory and anti‐oxidative effects of korean red ginseng extract in human keratinocytes. Immune Netw 2011;11:42–9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15. Kim HA, Kim S, Chang SH, Hwang HJ, Choi YN. Anti‐arthritic effect of ginsenoside Rb1 on collagen induced arthritis in mice. Int Immunopharmacol 2007;7:1286–91. [DOI] [PubMed] [Google Scholar]
  • 16. Leung KW, Pon YL, Wong RN, Wong AS. Ginsenoside‐Rg1 induces vascular endothelial growth factor expression through the glucocorticoid receptor‐related phosphatidylinositol 3‐kinase/Akt and beta‐catenin/T‐cell factor‐dependent pathway in human endothelial cells. J Biol Chem 2006;281:36280–8. [DOI] [PubMed] [Google Scholar]
  • 17. Park JS, Park EM, Kim DH, Jung K, Jung JS, Lee EJ, Hyun JW, Kang JL, Kim HS. Anti‐inflammatory mechanism of ginseng saponins in activated microglia. J Neuroimmunol 2009;209:40–9. [DOI] [PubMed] [Google Scholar]
  • 18. Kim YS, Cho IH, Jeong MJ, Jeong SJ, Nah SY, Cho YS, Kim SH, Go A, Kim SE, Kang SS, Moon CJ, Kim JC, Kim SH, Bae CS. Therapeutic effect of total ginseng saponin on skin wound healing. J Ginseng Res 2011;35:360–7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19. Lu ZQ, Dice JF. Ginseng extract inhibits protein degradation and stimulates protein synthesis in human fibroblasts. Biochem Biophys Res Commun 1985;126:636–40. [DOI] [PubMed] [Google Scholar]
  • 20. Song KC, Chang TS, Lee H, Kim J, Park JH, Hwang GS. Processed Panax ginseng, sun ginseng increases type I collagen by regulating MMP‐1 and TIMP‐1 expression in human dermal fibroblasts. J Ginseng Res 2012;36:61–7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21. Lee J, Jung E, Lee J, Huh S, Kim J, Park M, So J, Ham Y, Jung K, Hyun CG, Kim YS, Park D. Panax ginseng induces human type I collagen synthesis through activation of Smad signaling. J Ethnopharmacol 2007;109:29–34. [DOI] [PubMed] [Google Scholar]
  • 22. Cho JS, Moon YM, Um JY, Moon JH, Park IH, Lee HM. Inhibitory effect of ginsenoside Rg1 on extracellular matrix production via extracellular signal‐regulated protein kinase/activator protein 1 pathway in nasal polyp‐derived fibroblasts. Exp Biol Med 2012;237:663–9. [DOI] [PubMed] [Google Scholar]
  • 23. Liang HC, Chen CT, Chang Y, Huang YC, Chen SC, Sung HW. Loading of a novel angiogenic agent, ginsenoside Rg1 in an acellular biological tissue for tissue regeneration. Tissue Eng 2005;11:835–46. [DOI] [PubMed] [Google Scholar]
  • 24. Yang N, Chen P, Tao Z, Zhou N, Gong X, Xu Z, Zhang M, Zhang D, Chen B, Tao Z, Yang Z. Beneficial effects of ginsenoside‐Rg1 on ischemia‐induced angiogenesis in diabetic mice. Acta Biochim Biophys Sin (Shanghai) 2012;44:999–1005. [DOI] [PubMed] [Google Scholar]
  • 25. Morisaki N, Watanabe S, Tezuka M, Zenibayashi M, Shiina R, Koyama N, Kanzaki T, Saito Y. Mechanism of angiogenic effects of saponin from ginseng Radix rubra in human umbilical vein endothelial cells. Br J Pharmacol 1995;115:1188–93. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 26. Sengupta S, Toh SA, Sellers LA, Skepper JN, Koolwijk P, Leung HW, Yeung HW, Wong RN, Sasisekharan R, Fan TP. Modulating angiogenesis: the yin and the yang in ginseng. Circulation 2004;110:1219–25. [DOI] [PubMed] [Google Scholar]
  • 27. Kimura Y, Sumiyoshi M, Kawahira K, Sakanaka M. Effects of ginseng saponins isolated from Red Ginseng roots on burn wound healing in mice. Br J Pharmacol 2006;148:860–70. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 28. Chan LS, Yue PY, Wong YY, Wong RN. MicroRNA‐15b contributes to ginsenoside‐Rg1‐induced angiogenesis through increased expression of VEGFR‐2. Biochem Pharmacol 2013;86:392–400. [DOI] [PubMed] [Google Scholar]
  • 29. Cheung LW, Leung KW, Wong CK, Wong RN, Wong AS. Ginsenoside‐Rg1 induces angiogenesis via non‐genomic crosstalk of glucocorticoid receptor and fibroblast growth factor receptor‐1. Cardiovasc Res 2011;89:419–25. [DOI] [PubMed] [Google Scholar]
  • 30. You HJ, Han SK. Cell therapy for wound healing. J Korean Med Sci 2014;29:311–9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 31. Werner S, Krieg T, Smola H. Keratinocyte‐fibroblast interactions in wound healing. J Invest Dermatol 2007;127:998–1008. [DOI] [PubMed] [Google Scholar]
  • 32. Han SK. Advances in wound repair. Korea: Koonja Publishing Inc, 2013. [Google Scholar]
  • 33. Wu HT, Chen XX, Xiong LJ. Experimental study of proliferation of Schwann cells cultured with ginsenoside Rb1. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi 2003;17:26–9. [PubMed] [Google Scholar]
  • 34. Xie XS, Liu HC, Fan JM, Li HJ. Effects of ginsenoside Rb1 on TGF‐beta1 induced p47phox expression and extracellular matrix accumulation in rat renal tubular epethelial cells. Sichuan Da Xue Xue Bao Yi Xue Ban 2009;40:106–10. [PubMed] [Google Scholar]
  • 35. Shergis JL, Zhang AL, Zhou W, Xue CC. Panax ginseng in randomised controlled trials: a systematic review. Phytother Res 2013;27:949–65. [DOI] [PubMed] [Google Scholar]
  • 36. Coon JT, Ernst E. Panax ginseng: a systematic review of adverse effects and drug interactions [Review]. Drug Saf 2002;25:323–44. [DOI] [PubMed] [Google Scholar]

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