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Σάββατο 27 Απριλίου 2019

Ethnopharmacology

Cytotoxic effect of aqueous ethanolic extract of Luffa cylindrica leaves on cancer stem cells CD44+/24- in breast cancer patients with various molecular sub-types using tissue samples in vitro

Publication date: 28 June 2019

Source: Journal of Ethnopharmacology, Volume 238

Author(s): Ibrahim Mohamed Abdel-Salam, A.A. Abou-Bakr, Mohamed Ashour

Abstract

Ethnopharmacological relevance: Luffa cylindrica (L.) M.Roem is a climbing plant its parts have been used as traditional medicine for the treatment of different types of diseases including diarrhea, inflammation, cancer and viral infections. The parts used include fruit, seeds and leaves.

Aim of the study: Our study aims to investigate the effect of the aqueous-ethanol extract of Luffa cylindrica leaves on breast cancer stem cells CD44+/24- and other cell sub-populations using clinical samples with different molecular sub-types of breast cancer in vitro.

Materials and Methods: Breast tissues were obtained from patients undergoing surgery for the removal of breast tumors after complete clinical and pathological investigations. Tissue samples were processed to cell suspensions and treated with the extract in the tissue culture laboratory. Percentages of cell sub-populations within tumors and viability were measured by flowcytometry using clusters of differentiation as cell markers.

Results

Our results revealed that there were decreases in the total cell viability, CD44+/24- and total CD24+ cell sub-populations percentages after treatment with the extract, this may be an important indication of using Luffa leaves extract in the treatment of breast cancer or in combination with the traditional treatments.

Conclusion: Luffa cylindrica has proven to have anticancer activity on three different subtypes of breast cancer including luminal A, luminal B and Her2/neu enriched more over it has cytotoxic effect on both bulk tumor cells as well as cancer stem cells sub population CD44+/24- which possess high tumorigenic potency, these results were confirmed by measuring their viable number after treatment and sphere formation assay results.

Graphical abstract

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The new buzz: Investigating the antimicrobial interactions between bioactive compounds found in South African propolis

Publication date: 28 June 2019

Source: Journal of Ethnopharmacology, Volume 238

Author(s): K. Kharsany, A. Viljoen, C. Leonard, S. van Vuuren

Abstract
Ethnopharmacological relevance

Propolis, a resinous substance produced by the Apis mellifera bee, contains a number of flavonoids sourced from plants found in the surrounding region. Whilst bees use this substance to seal off and protect the beehive, humans have used propolis therapeutically for centuries, making use of its antibacterial, antiseptic, antipyretic and wound healing properties, among others. South African propolis is rich in the flavonoids pinocembrin, galangin, and chrysin and very little previous research has been conducted on the antimicrobial effects of these compounds.

Aim of the study

To obtain an understanding of the antimicrobial activity of the compounds pinocembrin, galangin, and chrysin, both independently and in combination.

Materials and methods

The compounds pinocembrin, galangin and chrysin were investigated for interactive antimicrobial activity by determining the minimum inhibitory concentrations (MIC), minimum bactericidal concentrations (MBC), anti-quorum sensing activity, biofilm studies, and toxicity studies (brine shrimp lethality assay).

Results

Minimum inhibitory concentration results demonstrated that combinations of compounds showed better inhibitory activity than single compounds. When the flavonoids were tested in combination using the MIC assay, synergy was noted for 22% of the 1:1 ratio combinations and for 66% of the triple 1:1:1 ratio combinations. Similarly, MBC results showed bactericidal activity from selected combinations, while the compounds on their own demonstrated no cidal activity. Quorum sensing studies showed that compound combinations are more effective at inhibiting bacterial communication than the individual compounds. Biofilm assays showed that the highest percentage inhibition was observed for the triple combination against E. coli at 24 h. Finally, brine shrimp lethality studies revealed that combinations of the three compounds had reduced cytotoxicity when compared to the individual compounds.

Conclusion

The results obtained in this study demonstrate that the compounds found in South African propolis work synergistically to achieve an optimal antimicrobial effect, whilst simultaneously minimizing cytotoxicity.

Graphical abstract

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Self-reported prevalence and severity of opioid and kratom (Mitragyna speciosa korth.) side effects

Publication date: 28 June 2019

Source: Journal of Ethnopharmacology, Volume 238

Author(s): Azlin Saref, Sharifah Suraya, Darshan Singh, Oliver Grundmann, Suresh Narayanan, Marc T. Swogger, Walter C. Prozialeck, Edward Boyer, Nelson Jeng Yeou Chear, Vicknasingam Balasingam

Abstract
Ethnopharmacological relevance

Mitragyna speciosa (Korth.) is a traditional medicinal plant widely used in Southeast Asia for its opioid-like effects. Although kratom produces analgesia through binding of mitragynine and other alkaloids at the mu-opioid receptor (MOR), the association of long-term kratom use with adverse opioid-like effects remains unknown.

Aim of the study: To determine the self-reported prevalence and severity of opioid-related adverse effects after kratom initiation in a cohort of illicit opioid users.

Materials and methods

A total of 163 illicit opioid users with current kratom use history were recruited through convenience sampling from the northern states of Peninsular Malaysia. Face-to-face interviews were conducted using a semi-structured questionnaire.

Results

Respondents were all males, majority Malays (94%, n = 154/163), with a mean age of 37.10 years (SD = 10.9). Most were single (65%, n = 106/163), had 11 years of education (52%, n = 85/163) and employed (88%, n = 144/163). Half reported using kratom for over >6 years (50%, n = 81/163), and 41% consumed >3 glasses of kratom daily (n = 67/163). Results from Chi-square analysis showed kratom initiation was associated with decreased prevalence of respiratory depression, constipation, physical pain, insomnia, depression, loss of appetite, craving, decreased sexual performance, weight loss and fatigue.

Conclusions

Our findings indicate that kratom initiation (approximately 214.29 mg of mitragynine) was associated with significant decreases in the prevalence and severity of opioid adverse effects.

Graphical abstract

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Comprehensive chemical profiling of monascus-fermented rice product and screening of lipid-lowering compounds other than monacolins

Publication date: 28 June 2019

Source: Journal of Ethnopharmacology, Volume 238

Author(s): Jin-Xiu Liang, Qun-Qun Zhang, Yan-Fei Huang, Han-Qing Pang, Xin-Guang Liu, Wen Gao, Ping Li, Hua Yang

Abstract
Ethnopharmacological relevance

Monascus-fermented rice product (MFRP) has been regarded as a dietary supplement and traditional medicine with circulation-promoting effects in China and other countries for centuries.

Aim of the study

This study was carried out to profile the chemical components in MFRP, and provide available information for elucidating the potential lipid-lowering compounds other than monacolins.

Materials and methods

High-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (HPLC-QTOF MS) and gas chromatography coupled with mass spectrometry (GC-MS) methods were applied to comprehensive analysis of chemical components in MFRP. Potential small molecules were identified by comparing with reference standards, or tentatively characterized by comparing their retention time and high-resolution mass spectral data with previous literature. The lipid-lowering properties of ten major non-monacolin compounds were evaluated in cholesterol-fed zebrafish larvae. And one with optimum lipid-lowering activity was subsequently evaluated in high fat diet-fed C57BL/6 J mice, with the dyslipidemia and ectopic lipid deposition being investigated.

Results

A total of 99 compounds were characterized in MFRP, including 38 monacolins, 5 decalins, 6 isoflavones, 13 pigments, 8 azaphilonoids, 11 amino acids, 4 nucleosides, 9 lipid acids, 4 phytosterols and glycerol. The preliminary screening showed that ergosterol remarkably reduced cholesterol levels in zebrafish larvae. Moreover, ergosterol delayed body weight gain and decreased circulating total cholesterol, triglyceride, low density lipoprotein cholesterol levels in high fat diet-fed mice. Ectopic lipid accumulation was also ameliorated in the liver and heart of obese mice.

Conclusion

Global analysis of chemical components and screening of lipid-lowering non-monacolin compounds in MFRP have improved our understanding of its therapeutic material basis.

Graphical abstract

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Ameliorative effect of Atractylodes macrocephala essential oil combined with Panax ginseng total saponins on 5-fluorouracil induced diarrhea is associated with gut microbial modulation

Publication date: 28 June 2019

Source: Journal of Ethnopharmacology, Volume 238

Author(s): Jing Wang, Wuwen Feng, Shiyang Zhang, Lu Chen, Yongcheng Sheng, Fei Tang, Junlin He, Xin Xu, Hui Ao, Cheng Peng

Abstract
Ethnopharmacological relevance

Traditional Chinese medicine (TCM) holds that deficiency of spleen-Qi is the major pathogenesis of chemotherapy-induced diarrhea (CID). Herb pair of Atractylodes macrocephala Koidz. (AM) and Panax ginseng C. A. Mey. (PG) has good effects of supplementing Qi and strengthening spleen.

Aim of the study

To investigate therapeutic effects and mechanism of Atractylodes macrocephala essential oil (AMO) and Panax ginseng total saponins (PGS) alone and in combination (AP) on 5-fluorouracil (5-FU) chemotherapy induced diarrhea in mice.

Materials and methods

The mice were administered with AMO, PGS and AP respectively for 11 days, and intraperitoneally injected with 5-FU for 6 days since the 3rd day of the experiment. During the experiment, the body weights and diarrhea scores of mice were recorded daily. Thymus and spleen indexes were calculated after sacrifice of the mice. Pathological changes in ileum and colonic tissues were examined by hematoxylin-eosin (HE) staining. And the content levels of intestinal inflammatory cytokines were measured by enzyme-linked immmunosorbent assays (ELISA). 16S rDNA Amplicon Sequencing was used to analyze and interpret the gut microbiota of fecal samples.

Results

AP significantly inhibited body weights loss, diarrhea, reductions of thymus and spleen indexes, and pathological changes of ileums and colons induced by 5-FU. Neither AMO nor PGS alone significantly improved above-mentioned abnormalities. Besides, AP could significantly suppressed the 5-FU-mediated increases of the intestinal inflammatory cytokines (TNF-α, IFN-γ, IL-6, IL-1β and IL-17), while AMO or PGS only inhibited some of them after 5-FU chemotherapy. Gut microbiota analysis indicated that 5-FU induced overall structural changes of gut microbiota were reversed after AP treatment. Additionally, AP significantly modulated the abundances of different phyla similar to normal values, and restored the ratios of Firmicutes/Bacteroidetes (F/B). At genus level, AP treatment dramatically decreased potential pathogens like BacteroidesRuminococcusAnaerotruncusand Desulfovibrio. AP also antagonized the abnormal effects of AMO and PGS alone on certain genera like BlautiaParabacteroides and Lactobacillus. Neither AMO nor PGS alone inhibited changes of gut microbial structure caused by 5-FU.

Conclusions

AP, combination of AMO and PGS, not AMO or PGS alone, significantly ameliorated diarrhea, inhibited intestinal pathology, and modulated gut microbial structure in 5-FU induced mice. AP also antagonized abnormal effects of AMO or PGS on certain genera. The results illustrated that gut microbiota was involved in the combined effects of AP on 5-FU induced diarrhea.

Graphical abstract

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Analgesic and anti-inflammatory effects and molecular mechanisms of Kadsura heteroclita stems, an anti-arthritic Chinese Tujia ethnomedicinal herb

Publication date: 28 June 2019

Source: Journal of Ethnopharmacology, Volume 238

Author(s): Huang-he Yu, Ye Lin, Rong Zeng, Xin Li, Ting Zhang, Shumaila Tasneem, Cong Chen, Yi-xing Qiu, Bin Li, Jin Liao, Yu-hong Wang, Xiong Cai, Wei Wang

Abstract
Ethnopharmacological relevance

Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by failure of spontaneous resolution of inflammation. The stem of Kadsura heteroclite (KHS) is a well-known anti-arthritic Tujia ethnomedicinal plant, which named Xuetong in folk, has long been used for the prevention and treatment of rheumatic and arthritic diseases.

Aim of the study

The analgesic and anti-inflammatory effects and the potential mechanisms behind such effects of KHS would be investigated by using different animal models.

Materials and methods

The abdominal writhing episodes of mice induced by intraperitoneal injection of acetic acid and the tail-flick response induced by radiant heat stimulation were used to evaluate the analgesic effect of KHS. The number of abdominal writhing episodes of mice and the latency of tail-flick in rats were measured and recorded. In acute inflammatory models, the ear edema of mice was induced by applying xylene on the ear surface, while the paw edema of male and female rats was induced by subcutaneous injection of carrageenan into the right hind paws of animals. The carrageenan-induced paw swelling in rats were selected as an anti-acute inflammatory mechanism of KHS. Serum levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor (TNF-α) were measured by ELISA, and protein expression of cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) were detected by Western blot.

Results

The maximal tolerated single dose of KHS was determined to be 26 g/kg in both sexes of mice. Pharmacological studies showed that KHS at the dose of 200 mg/kg significantly prolonged the reaction time of rats to radiant heat stimulation and suppressed abdominal writhing episodes of mice induced by intraperitoneal injection of acetic acid. KHS at the dose of 200, 400, and 800 mg/kg, showed dose-dependent inhibition of xylene-induced ear swelling in mice. KHS at the dose of 100, 200, 400, and 800 mg/kg demonstrated dose- and time-dependent suppression of paw edema induced by subcutaneous injection of carrageenan in both all rats. Mechanistic studies revealed that the anti-inflammatory effect of KHS was associated with inhibition of the production of pro-inflammatory cytokines IL-1β, IL-6, and TNF-α and effectively decreased the expression of COX and iNOS proteins in the carrageenan-injected rat serum, paw tissues and inflammatory exudates. The positive reference drug, rotundine at a dosage of 100 mg/kg and indomethacin at a dosage of 10 mg/kg were used in both mice and rat models.

Conclusion

These results suggested that KHS has significant effects on analgesia and anti-inflammation with decreasing the pro-inflammation cytokines of IL-1β, IL-6, and TNF-α and inhibiting the proteins expression of COX-2 and iNOS.

Graphical abstract

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The effect of Xiang-Sha-Liu-Jun-Zi tang (XSLJZT) on irritable bowel syndrome: A randomized, double-blind, placebo-controlled trial

Publication date: 28 June 2019

Source: Journal of Ethnopharmacology, Volume 238

Author(s): Yi-Sing Shih, Chang-Hai Tsai, Tsai-Chung Li, Hsueh-Chou Lai, Kun-Teng Wang, Wen-Ling Liao, Ching-Liang Hsieh

Abstract
Background

Irritable bowel syndrome (IBS) is characterized by chronic, recurrent abdominal pain or abdominal discomfort and changes in defecation habits. Xiang-Sha-Liu-Jun-Zi tang (XSLJZT) is a traditional Chinese medical formula that can modulate gastrointestinal disturbance.

Purpose

To investigate the therapeutic effect of XSLJZT on IBS.

Materials and methods

This study was designed as a double-blind, randomized, controlled preliminary study. Eighty patients with IBS were assigned to a control group (CG, 40 patients) that received oral administration of an XSLJZT 10% comparator (3.0 g three times daily) for 28 days or to a treatment group (TG, 40 patients) that received XSLJZT. The primary outcome measure was changes in scores on the Gastrointestinal System Rating Scale-IBS. The secondary outcome measures were changes in scores on the irritable bowel syndrome-quality of life (IBS-QOL) and World Health Organization-quality of life-brief.

Results

A total of 63 patients completed the study (n = 31 for the CG; n = 32 for the TG). The TG were discovered to have significantly lower diarrhea scores than the CG at V2 (second assessment) compared with V1 (first assessment, baseline) (mean change ± SD: CG: 0.19 ± 1.33 vs. TG: −0.38 ± 0.91, p = 0.05). The scores for Item 28 on the IBS–quality of life (QOL) scale ("I feel frustrated that I cannot eat when I want because of my bowel problems") were lower in the CG at V3 (third assessment) compared with V1 but slightly higher in the TG (CG: −0.48 ± 0.89 vs. TG: 0.03 ± 0.65, p = 0.01).

Conclusion

Oral administration of XSLJZT (3.0 g) for 28 days lowered the mean diarrhea score in patients with IBS, indicating that the patients in the TG had greater diarrhea improvement than those in the CG. The present study used 10% XSLJZT as a comparator, and the different items of the Gastrointestinal System Rating Scale–IBS, IBS-QOL, and World Health Organization Quality of Life–Brief were scored separately. Therefore, the selection of an appropriate comparator or placebo and score assessment are crucial issues for future study.

Graphical abstract

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Lonicera japonica extract increases metformin distribution in the liver without change of systemic exposed metformin in rats

Publication date: 28 June 2019

Source: Journal of Ethnopharmacology, Volume 238

Author(s): Seung Yon Han, Hee-Sung Chae, Byoung Hoon You, Young-Won Chin, Hojun Kim, Han Seok Choi, Young Hee Choi

Abstract
Ethnopharmacological relevance

Flower and flower bud of Lonicera japonica, Lonicerae Flos, have been popularly used as medicinal plant for the treatment of clearing heat and thirst, thereby improving diabetic or diabetic associated symptoms (thirst and poor eyesight).

Aim of the study

Organic cation transporters (OCTs) and multi-drug and toxin extrusion proteins (MATEs) are known to play important roles in metformin transport in the liver and kidneys. Thus, there might be interactions between Lonicerae Flos and metformin via OCTs and MATEs. Also treatment period has been issued in transporter-mediated drug interactions. The objective of this study was to determine the effect of Lonicerae Flos ethanol extract (LJ) on metformin pharmacokinetics and its glucose lowering activity in different treatment periods.

Materials and methods

Effect of LJ on metformin uptake was evaluated in vitro HEK-293 cells expressing human OCTs or MATEs. Treatment period-dependent impact of LJ on systemic exposure and hepatic distribution of metformin as well as its glucose tolerance activity were assessed in in vivo rats.

Results

LJ substantially inhibited MATE1-mediated metformin uptake in vitro. In evaluating treatment period effects of LJ and metformin, 1-, 7-, and 28-day co-treatments of LJ with metformin did not change systemic exposure of metformin compared to those in metformin alone. Whereas, 28-day co-treatment of LJ with metformin increased metformin concentration in liver as a pharmacological target site of metformin. It could be due to the reduced MATE1-mediated metformin efflux from hepatocytes to bile by MATE1 inhibition in liver. Glucose tolerance activity was also enhanced by 28-day co-treatment of LJ and metformin compared to metformin alone.

Conclusions

In 28-day co-treatment of LJ and metformin, LJ increased metformin concentration in liver and improved glucose tolerance activity without systemic exposure change of metformin, suggesting the importance to consider treatment period effect and both systemic exposure and tissue distribution in drug interactions.

Graphical abstract

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Dahuang Zhechong Pill suppresses colorectal cancer liver metastasis via ameliorating exosomal CCL2 primed pre-metastatic niche

Publication date: 28 June 2019

Source: Journal of Ethnopharmacology, Volume 238

Author(s): Chunhui Chen, Xueqing Yao, Yihua Xu, Qingyuan Zhang, Hao Wang, Liang Zhao, Ge Wen, Yawei Liu, Linlin Jing, Xuegang Sun

Abstract
Ethnopharmacological relevance

Dahuang Zhechong Pill (DZP) is a classical formula from "Synopsis of Prescriptions of the Golden Chamber". It has been used for treatment of abdominal masses (including tumorous diseases) for thousands of years.

Aim of the study

Our previous work showed that DZP suppresses CCl-4 induced hepatic fibrosis by downregulating the expression of interleukin-13. We aimed to test if DZP suppresses the metastasis of colorectal cancer (CRC) by ameliorating the fibrosis status of the future metastatic organ.

Materials and methods

Liver metastasis was observed by injection of MC38-EGFP cells with stably expressing enhanced green fuorescence protein beneath the splenic capsule of C57BL/6J mice. MC38-EGFP-derived exosomes were analyzed by Label-free comparative proteomics. mRNA expression was determined by Quantitative PCR. Protein expression was determined by immunohistochemistry, immunofuorescence and Western blot. Collagen deposition was determined by Masson staining. All data were statistically analyzed using SPSS.

Results

DZP drastically reduced the metastatic tumor number and fluorescence intensity in a splenic liver metastasis model. It also lowered the expression of mature TGF-β1 and decreased the fibronectin contents & collagen deposition. Exosome proteomics showed that the upregualted CC chemokine ligand-2 (CCL2) was repressed by DZP treatment. Importantly, DZP markedly lowered the expression of CCL2 and its receptor CCR2 in the liver. Exosomal CCL2 activated macrophage recruitment and shifted the M1/M2 paradigm to a M2 phenotype. DZP reduced the macrophage infiltration and attenuated the M2 polarizaion in tumor-bearing mice liver. It decreased the F4/80 positive areas and specifically reduced the ratio of CCR2+ positive macrophage. Anti-fibrosis and inhibition of CCR2 suppress the growth and metastasis of CRC.

Conclusions

DZP inhibits the liver metastasis of CRC by suppressing CCL2 mediated M2-skewing paradigm and ameliorating the pro-fibrotic microenvironment.

Graphical abstract

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Long-term administration of Aspalathus linearis infusion affects spatial memory of adult Sprague-Dawley male rats as well as increases their striatal dopamine content

Publication date: 28 June 2019

Source: Journal of Ethnopharmacology, Volume 238

Author(s): Justyna Pyrzanowska, Izabela Fecka, Dagmara Mirowska-Guzel, Ilona Joniec-Maciejak, Kamilla Blecharz-Klin, Agnieszka Piechal, Ewa Wojnar, Ewa Widy-Tyszkiewicz

Abstract
Ethnopharmacological relevance

Everyday use of the herbal tea rooibos, produced from Aspalathus linearis (Brum.f) Dahlg. (Fabaceae) is customary in South Africa, a continuation of its historical use by indigenous people. Although evidence of its traditional indications is anecdotal, rooibos tea is regarded as a general health tea.

Aims of the study

Available contemporary research indicates to broad cell protective activity of rooibos focusing on its antioxidative, anti-inflammatory, anti-hyperglycaemic and antithrombotic features affecting metabolic syndrome, cardiovascular risk and neuroprotection. Nevertheless little is known about its impact on brain functions. The present experiment aimed to evaluate the possible behavioural and neurochemical effects of long-term oral administration of "fermented"" rooibos herbal tea (FRHT) infusions to adult male Sprague-Dawley rats.

Materials and methods

Infusions, prepared using 1, 2 and 4 g of "fermented"" (oxidised) A. linearis leaves for 100 ml of hot water, were characterised in terms of flavonoid content by ultra-high and high performance liquid chromatography (UHPLC-qTOF-MS, HPLC-DAD) and administered to rats as sole drinking fluid for 12 weeks. Spatial memory behaviour was assessed in a modified version of the Morris water maze. Dopamine, noradrenaline, serotonin and their metabolite levels (DOPAC, 3-MT, HVA, MHPG, 5-HIAA) were quantified in prefrontal cortex, hippocampus and striatum by HPLC-ECD. Body weight and blood glucose level were additionally estimated.

Results

All FRHT-treated rats showed improvement of long-term spatial memory defined as increased number of crossings over the previous platform position in SE quadrant of the water maze. It was not accompanied by excessive motor activity. Striatal dopamine and its metabolite 3-MT (3-methoxytyramine) levels were increased in treated rats. There were no differences in body weight gain between control and treated animals but blood glucose level was significantly lower in the latter ones.

Conclusion

The improvement of long-term memory in FRHT-treated rats and stimulating impact of FRHT on their dopaminergic striatal transmission support the wellness enhancing effect of rooibos tea, contributing to a better understanding of the neurological background of traditional habitual consumption of this herbal tea.

Graphical abstract

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Medicine by Alexandros G. Sfakianakis,Anapafseos 5 Agios Nikolaos 72100 Crete Greece,00302841026182,00306932607174,alsfakia@gmail.com,

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