IN VIVO ANTIPLASMODIAL ACTIVITY OF METHANOL ROOT EXTRACT OF CISSUS CORNIFOLIA (BAKER) PLANCH
Abstract
The plant Cissus cornifolia belongs to the family vitacea. It is traditionally employed in the treatment of many ailments including malaria. The root was cold macerated using 70% methanol while phytochemical analysis was carried out to determine the secondary metabolites using standard protocol. The median lethal dose was then determined by exploiting the OECD protocol. The antiplasmodial activity was evaluated in mice intraperetonially infected with chloroquine sensitive plasmodium berghei-berghei using suppressive, prophylactic, and curative experimental models. Cissus cornifolia root extract revealed the presence of alkaloids, flavonoids, steroids/terpenoids, cardiac glycosides, carbohydrates, phenols, anthraquinones, and tannins. The oral LD50 of crude extract in mice was estimated to be greater than 5000 mg/kg per body weight. Curative effect of crude extract at doses of 250 mg/kg, 500 mg/kg and 1000 mg/kg showed statistical dose dependent significant reduction in mean parasitemia with a p value of 0.001. The standard drugs artesunate and chloroquine offered 85% and 90% respectively, when compared with non-treated mice. The extract, significantly prolonged mean survival time of mice infected with the parasites when compared with the distilled water administered group (Negative). Chemosuppresive effect showed reduction in mean parasitemia with suppression in the extract, chloroquine and artesunates administered groups as compared with the distilled water group. Prophylactic activity showed percentage parasite clearance of 62%, 73% and 65% with the different doses of the extract and 84% clearance for pyrimethamine when compared with negative group. The findings revealed that the extract possess antiplasmodial activity thereby validating its uses in folkloric medicine.
References
Abdullahi, Ibrahim Doma, Mustapha Ibrahim Gudaji and Abdullahi Hamza Yaro (2024) B-Hematin Inhibition activity of the Methanol Leaf Extracts and Residual Aqueous Fraction of Ficus Asperifolia. FUDMA Journal of Science. Volume 8 (15-22). DOI: https://doi.org/10.33003/fjs-2024-0802-2161
Adebayo, J.O. And Krettlia, A.U. (2010). Potential Antimalarials from Nigerian Plants: A Review. Journal of Ethnopharmacology, 133: 289302. DOI: https://doi.org/10.1016/j.jep.2010.11.024
Adefolaju, Toyin (2011). The Dyanamic and Changing Structure of Traditional Healing System in Nigeria. International Journal of Health Research: 4(2). 99-106.
Adesina, K. (2008). Traditional Medicine in Nigeria. Accessed July, 2018. http://www.OnlineNigeria.com/health
Adoum, O.A (2016). Screening of Medicinal Plant Native to Kano and Jigawa States of Northern Nigeria using Artem Cyst (Brine Shrimp Test). American Journal of Pharmacological Science,4: 3273-2344
Afisulahi, Abiodun, Yakubu Babatunde, Attahiru Al-Hajj Muhhamd, and Mark Devine (2022). Prevalence and Determinant of Herbal Medicine use among Adult Attending the National Health Insurance Clinic of Abubakar Tafawa Balewa University Teaching Hospital, Bauchi, Nigeria. Nigerian Journal of Medicine. Volume 31, 399-418. DOI: https://doi.org/10.4103/NJM.NJM_41_22
Aminake, N., Makoah and Pradel Gabriele (2013). Antimalarial Drugs Resistance in Plasmodium Falciparum and the Current Strategies to Overcome Them. Institute of Molecular Biotechnology, RWTH Aachen University, 269-282.
Amorosa, L.F., Corballinic, G. And Coluzzib, M. (2005). Lessons Learned from Malaria Italy Past and Sub-Saharas Future. Health and Place, 11:65-73. DOI: https://doi.org/10.1016/j.healthplace.2004.06.003
Anthony Ashmi and Parija Subhash Chandra (2016). Review of Antimalarial Resistance. Tropical Parasitology,6 (1): 30-41. DOI: https://doi.org/10.4103/2229-5070.175081
Baelmans, R., Deharo E., Munaz V., Sauvain and Ginsburg H (2000). Experimental conditions for testing the inhibitory activity of chloroquine on the formation of hematin. Experimental Parasitology; 96:243-248. DOI: https://doi.org/10.1006/expr.2000.4558
Balli, S. (2023). Pyrexia: A review of its Pathophysilogy, Diagnosis and Management>Journal of medical Science.43(2).53-6.
Bannister, L. And Mitchell, G (2003). The in, Outs and Roundabouts of Malaria. Trends in Parasitology, 19: 209-213. DOI: https://doi.org/10.1016/S1471-4922(03)00086-2
Basima, A. and B. H., Albadarani,(2021). A Preliminary Study of Malaria Infection (Plasmodium Spp) In Iraq Live Stock. Egyptian Journal of Veterinary Science. Volume 52, 97-111. DOI: https://doi.org/10.21608/ejvs.2020.46641.1195
Berhan Mengiste (2008). Antiplasmodial Activity of Extracts from Dodonaea Angustifolia L.F. and Bersama Abyssinica Fresen Against Plasmodium Berghei iin Mice Model. Addis Ababa University Press, Ethopia, 20-44.
Berkowitz, B.A. (2004). B.G. Katzung. Basic and Clinical Evaluation of New Drugs. Lange Medical Books, Mc Graw-Hill Medical Publishing Division New York Chicago,67.
Bhattacharya, S., (2021). Flavonoids as potential antimalarial agents.: A review of their mechanism of action and efficacy. Phytomedicine, 85, 153-1551.
Boham AB, Kocipal AC. 1994. Flavonoid and condensed tannins from leaves of Hawaiian vaccininum vaticulum and vicalycinium. Pacific Sciences., 48: 458-463.
Bonday ZQ, Dhanasekaran S, Rangarajan PN, Padmanaban G (2000). Import of Host Delta-aminolevulinate dehydratase into the Malarial Parasite: Identification of a New Drug Target. Natural Medicines; 6:898903. DOI: https://doi.org/10.1038/78659
Borquaye, Lawrence, Macheal Konney, Edward Ntim Gasu, and Glora Doh (2020) Anti-inflammator and antioxidant activities of extract of Ressantia Indica, Cissus cornifolia and Grossseria Vinei. Cognet Biology Journal.6;178-187. DOI: https://doi.org/10.1080/23312025.2020.1785755
Bragga Cssio, Martins Antonio Camarga, Klein Wganer, Wilson Brano and Silva Monaco (2015). Side Effect of Chlroquine and Symptomsreduction in Malaria Endemic Area (Mancio Lima, Acre and Brazil. Interdisplinary Perspective on Infectious DOI: https://doi.org/10.1155/2015/346853
Burkill, H.M., (2000). The Useful Plants of West Tropical Africa. 2nd Edition. 4, Families M-R. Royal Botanic Gardens, Kew, Richmond, United Kingdom. 969.
Carlton, J.M., Das, A. And Escalante A.A (2013). Genomics Population Genetics and Evolutionary History of Plasmodium Vivax. Advanced Journal of Parasitology. 81: 203-222. DOI: https://doi.org/10.1016/B978-0-12-407826-0.00005-9
Chinappi, M. (2020). Cardiac glycoside from calotropis procera disrupt osmotic balance of fasciola gigantica. Journal of Ethopharmacoloy, 247,112-341.
Clark, I.A., And Cowden, W. B (2003). The Pathophysiology of Falciparum Malaria. Journal of Pharmacology and Therapeutics. 99(2): 221-260. DOI: https://doi.org/10.1016/S0163-7258(03)00060-3
Das, D.K., Mukhereje, R., Chakraborty, C (2015). Computational Microscopic Imaging for Malaria Parasite Detection: A Systemic Review. Journal of Microscopy. https://doi.org/10.1111/jmi.12270 Accessed December 2021). DOI: https://doi.org/10.1111/jmi.12270
Depinay, J. F. Franetich, A. C. And Gr Uner (2011). Inhibitory Effect of TNF-on Malaria Pre Erythrocytic Stage Development: Influence of Host Hepatocyte/Parasite Combinations, PloS One, 6, (3), Article ID E17464. DOI: https://doi.org/10.1371/journal.pone.0017464
Doma A. I and Yaro A. H (2024) In Vivo Antiplasmodial and Schizont Inhibition Activity of Methanol Stem Bark Extract of Ficus vallis-Choude. Dutse Journal of Pure and Applied Sciences DUJOPAS 10 (3c): 21-38, 2024 DOI: https://doi.org/10.4314/dujopas.v10i3c.3
Dorsey, G., Vlahos, J., Kamya, M. R., Staedke, S. G. And Rosenthal, P. J. (2003). Prevention of Increasing Rates of Treatment Failure by Combining Sulfadoxine-Pyrimethamine with Artesunate or Amodiaquine for The Sequential Treatment of Malaria. Journal of Infection, 188: 12311238. DOI: https://doi.org/10.1086/378523
Dwivedi, A., (2021). Flavonoids and Phenolic acid: A comprehensive Review on their antioxidant, anti-inflammatory and antimicrobial properties. Journal of Food Science. 86(5). S 1448-1463.
Evans, W.C., (2002). Trease and Evans Pharmacognosy, 15th edition. W.R., Saunders Company Ltd, London, 15-77.
Felhi Samir, Daouda Amal Hajloui Hafedh, Gharsallah Neji,And Khadri Adel (2017). Solvent Extraction Effect On Phytochemical Constituent Profiles, Antioxidant And Antimicrobial Activities And Functional Group Analysis Of Ecballium Elatrium Seed And Peels Fruits. Journal of Food Science Technology, Campinas. 37(3), 483-492. DOI: https://doi.org/10.1590/1678-457x.23516
Ferreira, L. S., (2019). Flavonoids: A Review of their bioavailability, Metabolism and Health benefit. Nutrients, 11(22). 5218.
Francis, E.G. Cox (2010). History of the Discovery of the Malaria Parasites and their Vectors. Department of Infectious and Tropical Diseases. London School of Hygiene and Tropical Medicine, London WC1E 7HT UK. 230-245.
Fulata, M., Ali Hamidu, Usman, Arastud William, Sule, Ibrahim and Adamu I., Zadva (2017). Phytochemical, Analgesic and Anti-Imflammatory Evaluation of the Crude Ethanol and N. Hexaneextract of The Aerial Part of Laggera Aurita Linn. Chemistry Research Journal, 2(6) :22-28.
Gogoi, P. (2021). Limonoids. A comprehensive review on their chemistry, biosynthesis and Pharmacological activities. Phytochemistry Review, 20(2), 257-278.
Goldberg DE. (1993). Hemoglobin degradation in Plasmodium-infected red blood cells. Semin Cell Biology; 4:35561. DOI: https://doi.org/10.1006/scel.1993.1042
Greenwood, B.M. (2008). Control to Elimination: Implications for Malaria Research. Trends in Parasitology. 24: 450-454. DOI: https://doi.org/10.1016/j.pt.2008.07.002
Hansen D.S (2012). Inflammatory Responses Associated with The Induction of Malaria: Lessons from Experimental Murine Models. PloS Pathogens, 8: E1003045. DOI: https://doi.org/10.1371/journal.ppat.1003045
Harbone J.B (1973). Phytochemical methods. London. Chapman and Hall, Ltd., 49-188.
Hawa Mbali, Jessie JaneKhaki, and Alinane, Linda (2021). Prevelence and Correlate of Herbal Medicine use among Anti- Retro Viral Therapy (ART) Client at Queen Elizabeth Central Hospital (QECH), Blantyre Malawi: Cross-Sectional Study. Malawi Medical Journal. 33(3).153-158 DOI: https://doi.org/10.4314/mmj.v33i3.2
Hill, R.D., J.K. Baird, M.E. Parise, L.S. Lewis, E.T. Ryan, And A. J. Magill (2006). Primaquine: Report from CDC Expert Meeting Onmalaria Chemoprophylaxis, The American Journal of Tropica lmedicine and Hygiene, 75 (3): 402415. DOI: https://doi.org/10.4269/ajtmh.2006.75.402
Hyde, J. E. (2007). DrugResistant Malaria an Insight. FEBS Journal. 274(18), 4688-4698. DOI: https://doi.org/10.1111/j.1742-4658.2007.05999.x
Idris, M.l., nkkafamiya, I.I., Akinterinwa,A., and Japari, J.I (2015). Preliminary studies of some medicinal plants in Girei, Adamawa state of Nigeria. British Journal of Pharmaceutical research. 6(3).203-213. DOI: https://doi.org/10.9734/BJPR/2015/16205
Idro, R., Jenkins, N. E., And Newton, C. R. (2005). Pathogenesis, Clinical Features, And NeurologicalOutcome of Cerebral Malaria. The Lancet Neurolog,. 4(12), 827-840. DOI: https://doi.org/10.1016/S1474-4422(05)70247-7
Jimoh A, Tanko Y, Mohammed A (2013). Modulatory role of methanolic leaf extract of Cissus cornifolia on blood glucose levels of normoglycemic wistar rats. European Journal of Experimental Biology, 3:; 227.
Jude, O.K (2017). Secondary Metabolites from Plant.: A Review of their Antimicrobial, antoxidants, anticancer Properties. Journal of Pharmacy and Pharmacology, 74(1),11-34.
Karande AM, Kamble HV., Kumbhar VH., Kane SR., Maghdum CS. (2016), Preliminary Phytochemicals Profile and Antibacterial screening of Glochidion elliptecium. European journal of Experimental Pharmacology. ;6(4).41-45
Katerikul Surasak, Buranapong Jirayut, Sriturvanich Wareyut and Kaewthamsorn (2016). The Development of Malaria Diagnostic Techniques: A Review of the Approaches with Focus on Dieelectricphoretic and Magnophoretic Methods. Malaria Journal. 15: 358-369. DOI: https://doi.org/10.1186/s12936-016-1400-9
Katzung, B.G., Masters, S.B., And Trevor, A.J. (2012). Basic And Clinical Pharmacology. New York: Mcgraw-Hill Medical.12th Edition.
Kwambe, B. A., Okoro, E. U and Okoro CS (2019). Antiplasmodial activity of Artemesia annua aqueous extracts in plasmodium berghei berghei infected mice. Journal of Ethnopharmacology, 231, 145-153.
Liu Ying, Zhou Rui-Min, Zhang Ya-Lan, Wang DUO-Quan, Li SU-Hua, Yang Cheng-Yun, Qlan Dan, Zhoa Yu-Lin, Zhang Hong-Wei and Xu Bian (2018). Analysis Polymorphisms in The Circumsprozoite Protein Gene of Plasmodium vivax Isolate from Henan Province, China. Malaria Journal,17(103): 234-242. DOI: https://doi.org/10.1186/s12936-018-2237-1
Lorke D. A new approach to practical acute toxicity testing. Archives of Toxicology, 54: 1983; 275-87 DOI: https://doi.org/10.1007/BF01234480
Maigemu Ahmad Yahaya and Kalthum Haji Hassan (2015). Malaria as a Cause of Mortality and Morbidity: Socioeconomic Overview. Journal on Humanities and Social Science:5(8) 18-28.
Mehlhorn, H (2018). Cardiac glycosides disrupt integrity of membranes and the contractility of muscles in parasites. Parasitology Research, 117(10), 2931-2938.
Memede, O., W., Gwadhri, and P., Lars (2020). Phenolic compounds: A review of structural diversity, biosynthesis and Biological activities. Journal of food sciences, 85(5).
Musa, A.M., Yaro, A.H., Usman, H., Magaji, M.G. and Habu, M. (2008). Phytochemical and someneuropharmacological studies on the methanolic leaf extracts of Cissus cornifolia (Vitaceae) in mice. International Journal of Pharmacology; 4:145-148 DOI: https://doi.org/10.3923/ijp.2008.145.148
Nayyar, G.M., Breman, J.G., and Newton, P. N. (2012). Poor Quality Antimalarial Drugs in Southeast Asia and Sub Saharan Africa. Journal of Infectious Disease. Volume 12(7).506-520. DOI: https://doi.org/10.1016/S1473-3099(12)70064-6
Noulin F (2019). Oxidative stress in malaria: Double edge sword. Online journal accessed 20 th November, 2024. Doi 10.1002/1873.1352.
Obadoni BO, Ochuko PO. 2001. Phytochemical studies and Comparative efficacy of the crude extracts of some homeostatic plants in Edo and Delta States of Nigeria. Global Journal of Pure and Applied Sciences. 8: 203-208. DOI: https://doi.org/10.4314/gjpas.v8i2.16033
Olliaro P. L and Goldberg D. E. (1995). The plasmodium digestive vacuole: metabolic headquarters and choice drug target. Parasitology Today; 11:2947 DOI: https://doi.org/10.1016/0169-4758(95)80042-5
Olurokoba, A., O., (2018). Toxoplasmosis: A review on toxogenesis, Diagnosis and Control. Journal of Parasitic diseases. 42(3).537-547.
Olusegun, L., Thomas, R., And Micheal, I., M (2012). Curbing Aternal and Child Mortality. The Nigerian Experience: Doi:10:5897?INJM
Onaku, Linda O (2010). Evaluation of Combinations of Artesunic Acid and Aqueous Extract of Azadirachta Indica or Carica Papaya on the Reduction of Parasitemia in Mice Infected with Plasmodium berghe. University of Nigeria Nsukka Press, 5-126
Peters W (1967). Rational Methods in the Search for Anti-Malaria Drugs. Transient Research in Social Tropical Medecine. 61: 400-410. DOI: https://doi.org/10.1016/0035-9203(67)90015-6
Peters, W (1998). Drug Resistance in Malaria Parasites of Animals and Man. Advances in Parasitology, 41, 1-62. DOI: https://doi.org/10.1016/S0065-308X(08)60421-2
Peters, W. & Robinson, B. L. (1999). Handbook of Animal Models of Infection, London. Academic Press. Pharmacology, 74(1),11-34.
R. A. Turner, Screening Methods in Pharmacology, Academic Press, New York, NY, USA, 1965
Rayner C., Julian (2015). Plasmodium Malaria Malaria. EBIO Medicine. 2: 1023-1029. DOI: https://doi.org/10.1016/j.ebiom.2015.08.035
Ruqayyah, I. H. Nock, I., Nadams and B.D.J George (2017). Investigating Prevalence and Intensity of Plasmodium falciparum Infection and Two Method of Malaria Diagnosis in North Western Nigeria. International Journal of Tropical Disease. 23(2): 1-8. DOI: https://doi.org/10.9734/IJTDH/2017/28884
Russo, Gianluca, Giovanni Faggioni, Giacomo Maria Paganotti, Ghysaline Bruna Djeunang Dongho, Allce Pompanni, Ricardo Desentis, Gianpero Tebano, Mpoame Mbida, Martin Sanou Sobze, Vincenzo Vullo, Glovenni Rezza and Florig Romano Lista (2017). Molecular Evidence of Plasmodium vivax Infection in Duffy Negative Symptomatic Individuals from Dschang, West Cameroon. Malaria Jounal.16:74-87. DOI: https://doi.org/10.1186/s12936-017-1722-2
Ryley JF, Peters W (1970). The Anti-Malaria Activity of Some Quinolone Esters. American Tropical Medical Parasitology. 84: 209-222. DOI: https://doi.org/10.1080/00034983.1970.11686683
Safil Muheet Alam, Beg Tanveer, Harath Abdul Halim, Altayalan Fahad Suleman and Alquaish Saleh (2013). Antimalarial Drugs: Mode of Action and Status of Resistance. African Journal of Pharmacy and Pharmacology: 7(5), 148-156 DOI: https://doi.org/10.5897/AJPPX12.015
Shabbir, M., Muhammad, R. K. And Naima, S. (2013). Assessment of Phytochemicals, Antioxidant, Anti-Lipid Peroxidation and Antihemolytic Activity of Extract and Various Fractions of Mytenus Royleanus. BMC Complementary and Alternative Medicine. 13:143-145. DOI: https://doi.org/10.1186/1472-6882-13-143
Shen, Hai-Mo Shen-Bio Chen and Jun-Nhu (2015). Whole Genome Sequencing of a Plasmodium vivax Isolated from the ChinaMyanamar Border Area. Rio De Janeiro.110(6): 814-816. DOI: https://doi.org/10.1590/0074-02760150216
Sigala P. A and Goldberg D. E (2014). The Peculiarities and Paradoxes of Plasmodium Heme Metabolism. Annual Revision Microbiology; 68:25978. DOI: https://doi.org/10.1146/annurev-micro-091313-103537
Sinka Marianne E., Rubio-Palis Yasmin, Manguin Sylvie, Patil Anand P., Temperley Will H, Petter W Gettting, Thomas Van Boeckel, Kbaria W., Caroline, Harback E., Raph and Simon I Hay (2010). The Dominant Anopheles Vectors of Human Malaria in the Americas: Occurrence Data, Distribution Map and Bionomic Precis. Parasite and Vectors. 3 (72): 1-26. DOI: https://doi.org/10.1186/1756-3305-3-72
Sofowora, A. (2008). Medicinal Plants and Traditional Medicine in Africa. Spectrum Books Limited, Ibadan, Nigeria, 3rd Edition. Pp. 200-202.
Somzeet P, Chouldry, NSK, Jagannath PV, Dippiti KP, (2009). Analgesics, Antipyretic and Antiimflammatory effect of the whole plant extract of Desmotachya bipinnata stapf (Poaceac) in Albino rats. Drug Invent. 1(2).150-153.
Sundus Shafat Ahmad, Mnju Rahi, Vikash Ranjan and Amit Sharma (2021). Mefloquine as A Prophylaxis for Malaria Need to Be Revisited. International Journal of Parasitology. Dug and Resistance. Volume 17, 23-26. DOI: https://doi.org/10.1016/j.ijpddr.2021.06.003
Talent Chipiti, Muhammad Auwal Ibrahim, and Shahidul Islam (2017). In Vitro -Amylase and -Glucosidase Inhibitory and Cytotoxic Activities of Extract from Cissus Cornifolia Planch Parts. Journal of Pharmacognosy and Natural Products. Volume 13(50). 330-333 DOI: https://doi.org/10.4103/pm.pm_223_16
Tanko, Y., Kadiri, O.T., Mohammed, A., Mahdi, M.A. and Musa, K.Y. (2011). Preliminary anti-diarrhoeal activity of methanol extract of Cissus cornifolia (bak) planch on experimental animals. Annals of Biological Research; 2(4): 229-237.
Temesgen Bekele (2015). Invivo Anti-Malarial Evaluation of Leaf Extract of Vernonia amygdalina Del. (Asteraceae) Against Plasmodium berghei. Addis Ababa University Press, Ethiopia. 110-221
Trager W and Jansen JB. (1976). Human Malaria Parasites un continuous Culture. Science; 193 (4254): 673-5 DOI: https://doi.org/10.1126/science.781840
Trease, G.E. and Evans, M.C. (1983). Textbook of Pharmacognosy. Thirteenth Edition, Bailier Tindal, London, 247-762.
Tripathi, K. D (2003). Essential of Medical Pharmacology, 5th Edition, Jaypee Brothers Medical Publication (P) Ltd., New Delhi, 683-693.
Udobre A. S et al. (2013). Effect of Methanol Leaf Extract of Nauclea latifolia on Albino Mice Infected with Plasmodium berghei berghei. African Journal of Pharmacology and Therapeutics; 2 (3): 83 87
Walum, E (1998). Acute Toxicity of some naturally occurring compounds. Toxicology in vitro, 12 (2), 123-128.
WHO (2010). Traditionalmedicine Strategy. Geneva: Http://Apps.Who.Int/Iris/Handle/Accessed 20 November 2017.
WHO (2018). The Biology of Malaria Parasites: Report of WHO Scientific Group. WHO Technical Report Series 2. (Accessed On April, 2019)
Winter, R.W., Kelly, J.X., Smiikstein, M.J., Dodean, R., Bagby, G.C., Rathbun, R.K., Levin, J.I., Hinrichs, D. and Riscoe M.K (2006). Evaluation and Lead Optimization of Antimalarial Acridones. Journal of Pharmacology. 114:47-56. DOI: https://doi.org/10.1016/j.exppara.2006.03.014
World Health Organization (2022) Guidelines for the treatment of malaria. 2nd edition Geneva, Switzerland: WHO Press.
World Health Organization (WHO, 2020). Malaria Report.
World Health Organization (2017). World Malaria Report. Geneva. (Http://Creativecommons. Org/Licenses/By-Nc-Sa/3.0/Igo)
Yaro, A.H., Anuka, J.A., Salawu, O.A., Hussaini, I.M, Usman, H. and Musa, A.M. (2009). Comparative Neuropharmacological Activities Methanolic Extracts of Leaves and Roots of Cissus cornifolia in Mice. African Journal of Biomedical Research; 12(3): 219-223.
Zheinji, L. And Xu Chunbo (2011). The Fundamental Theory of Traditional Chinese Medicine and The Consideration in its Research Strategy. Journal of Frontiers of Medicine. 5(2): 208-211. DOI: https://doi.org/10.1007/s11684-011-0126-x
Copyright (c) 2025 FUDMA JOURNAL OF SCIENCES
This work is licensed under a Creative Commons Attribution 4.0 International License.
FUDMA Journal of Sciences
