IJRPC 2021, 11(1), 1-9 Athar Jamal et al. ISSN: 22312781

INTERNATIONAL JOURNAL OF RESEARCH IN PHARMACY AND CHEMISTRY

Available online at www.ijrpc.com Review Article DOI: https://dx.doi.org/10.33289/IJRPC.11.1.2021.11(8) A REVIEW ON PHARMACOLOGICAL ACTIVITIES OF RHAMNOIDES Athar Jamal*, Ayesha Maqsood and Riffat Ameen University of Management and Technology, Department of Chemistry, Johar Town, Lahore, Pakistan.

ABSTRACT Dependence on natural medicines in the administration of illnesses is still a lot of experienced by a huge portion of the individual particularly rustic inhabitants because of their simple availability and cost viability. Sea buckthorn (Hippophaerhamnoides L.) belongs to family , a one of important basically for its therapeutic and nourishing potential. Sea buckthorn (SBT) is a prickly nitrogen-fixing bush of cold dry climate confined to Europe and Asia. It is at present tamed in a few portions of the world because of its nourishing and therapeutic properties. The entire plant of sea buckthorn and particularly its berries are a wellspring of countless distinctive bioactive mixes. The best consideration has been attracted to its high substance of nutrients, minerals, common cancer prevention agents and proteins. Sea buckthorn is esteemed for its cardio protective, anti-atherogenic, anti-diabetic, hepatoprotective, hostile to cancer- causing, immunomodulatory, antioxidant, antiviral, antibacterial, mitigating and vasorelaxant impacts. This review sums up the pharmacological activities of Hippophaerhamnoides.

Keywords: Hippophaerhamnoides, pharmacological activities and medicinal herb.

INTRODUCTION practices, natural drugs are popular for Medicinal turned out to be an integral essential social insurance in developed as part of human progress to battle sicknesses well as underdeveloped nations. The from the earliest starting point of humankind. utilization of plants as a wellspring of Medicinal plants can be a significant prescriptions for the fix of numerous wellspring of recently known synthetics, with infections returns to ancient times and conceivable helpful impacts. Customary individuals from all continents have this old practice is the therapeutic utilization of practice. The medicinal properties of plants, a lot more established than regular therapeutic plants are expected to a great pharmaceutical, pharmacological, and extent to the presence of various complexes substance sciences. The world association synthetic compositional intensifies that exist has assessed that for its fundamental as secondary metabolites. Which are medicinal service needs; in excess of characterized into , alkaloids, seventy five percent of the world's population saponins, glycosides, starches, tannins and already relies upon plant-inferred drugs, basic oils. normally got from conventional healers. Due The word Hippophae has originated from a to their broad biological and therapeutic Latin word 'Hippo' which means pony and

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'Phaos' which signifies 'sparkles’ 1. detailed that Hippophaerhamnoides has Hippophaerhamnoides has a place with no cytotoxicity or unfavourable impact after family Elaeagnaceae usually known as Sea oral organization 2. The and organic buckhorn (SBT) 2. It is a group of of products have been utilized as germicide and the order. The family incorporates twisted mending just as in the treatment of around 100 in three genera for the ulcers in people asmedications in Turkey 6. most part found in the moderate geological Sea buckthorn seed can be utilized as oil and scopes of the Northern side of the equator. the leaves to make . While tea produced Sea berries are grouped in the plant scientific using the seeds have diuretic properties and classification into six species and 12 help in weight reduction, imbuements of the subspecies 3. Seaberries developed in the leaves have antidiarrheal properties; also, Czech Republic for the most part incorporate organic product reinforce the safe sea buckthorn and Russian silverberry. framework, and show action against skin Hippophaerhamnoides is a deciduous, problems. The positive and special properties dioecious plant with various greenish-yellow of sea buckthorn have been known since VII blossoms and brilliant orange, globular, Century BC. The plant was utilized in ellipsoid natural product. It is local to Europe, characteristic medication, yet in addition, in India, Nepal, Bhutan, Pakistan, France, veterinary medication as a methods for Afghanistan, Myanmar, , Romania, soothing helminthiasis in ponies and giving , and Britain 4. In india, them progressively mass and a delightful, Hippophaerhamnoides is broadly dispersed glossy coat. Now it is utilized in numerous in chilly Himalayan district at a height of businesses, particularly the pharmaceutical, 2500-4000m 2. Hippophaerhamnoides bush corrective, and food ventures, yet additionally is 2 meter tall with 2-6 cm long leaves. It is as an ornamental component, as firewood, or well developed all around in depleted soil even as an apparatus for the restoration of with pH 5.5-8.3. Hippophaerhamnoides plant ruined territories 7. As per authentic records, can persevere through aextraordinary sea buckthorn was first utilized as a temperature of 40-43ºC 4. medication in , and in increasingly The entire plant of Hippophaerhamnoides is current occasions, the plant was officially significant, berries are the most significant recorded in the Chinese Pharmacopeia in part from which the juice is removed 4. 1977 7. From 5 decades, Hippophaerhamnoides juice is a significant Hippophaerhamnoides has been utilized for wellspring of some important synthetic the treatment of harms caused by radiations, substances, for example, swelling and burns in Chinese traditional tocopherolmcrotrients, organic acids, vitamin medications8. Hippophaerhamnoides oils C and polyunsaturated fatty acids. The juice have been utilized in natruceuticals, regular of Hippophaerhamnoides was the regular prescriptions and beauty care products as medication utilized in antiquated. The berries crude materials in Russia for certain decades (fruits) of Hippophaerhamnoides are 9. Hippophaerhamnoides oil have additionally maturated in three stages. The first stage is been utilized in the treatment of skin issue, the quickening seed development, second for example, dermatitis, psoriasis, lupus stage is the declining progress seed erythematosus and dermatosus10. development and the last third stage is the Hippophaerhamnoides has been utilized in known as development stage. In most different areas of the world as customary region of the world including Pakistan the medication for the treatment of acid reflux, berries are aged toward the beginning of hack and blood sepsis 11. September. In the event that the branches At present, sea buckthorn has become well are not upset, at that point the berries stay known particularly for its constructive entire of the winter connected to the outcomes on the human living being. It branches 5. The natural products are known likewise diminishes stomach ulcers, to be a rich wellspring of nutrients, underpins wound mending, quickens carotenoids, flavonoids and sugar alcohols. treatment of skin issue and lessens torment The leaves contain a wonderful amount of 12-14. Sea buckthorn acts against asthma and supplements and bioactive substances, for aspiratory ailments 15, against expanded the most phenolic part. It was additionally sebum emission and influences platelet

2 IJRPC 2021, 11(1), 1-9 Athar Jamal et al. ISSN: 22312781 accumulation4. Its antistress and adaptogenic Antioxidant Activity exercises have been affirmed 14. Sea The antioxidant activity of buckthorn likewise emphatically influences Hippophaerhamnooides was determined by metabolic ailments 3, with the capacity to two techniques at different concentrations hinder maturing and ensure against ranging from 0.0625-0.5mg/ml. One is DPPH radiation-actuated harm, quicken the technique in which butylatedhydroxytoulene recuperating of consumes and frostbites, and (BHT) was used as reference standard and lessen balding. Sea buckthorn likewise second one is ABTS technique in which influences mental capacities, specifically trolox was used as a reference standard. IC50 diminishing memory misfortune in old values of both techniques were obtained. individuals. Its beneficial outcomes have IC50 estimation of fluid concentrate was seen been used for quickening of wound as essentially lower than BHT utilized in recovering, particularly in individuals after DPPH technique, while IC50 esteem by ABTS ear, nose and throat tasks where sea berry technique couldn't be determined as oil has been a part of the treatment. The oil concentrate showed very low antioxidant has been utilized for insurance against sun activity. IC50 estimations of MeOH remove based radiation 16. Sea berry items might be acquired by DPPH and ABTS technique were utilized in different medication structures from seen as fundamentally equivalent to the IC50 fluids through powder, pastes, patches, films, estimations of the particular reference treatments and mist concentrates to guidelines (BHT and Trolox). This suppositories 16. Hippophaerhamnoides plant recommended MeOH leaf extract have has impervious to dry season, cold, and antioxidant activity and is more dynamic than soluble base and salt. The perplexing root watery concentrate 19. framework with nitrogen-fixing of the In-vitro investigation of hydrophilic Hippophaerhamnoides an ideal pioneer plant concentrate of Hippophaerhamnoides in soil and water protection zone 9. Sea indicated great antioxidant activity, which buckthorn is a wellspring of natural acids, was like the antioxidant activity of the basically , quinicacid, oxalic acid, methanolic concentrate of oleic acid, citrus acid, and tartaric acid. Sea Hippophaerhamnoides20. The antioxidant buckthorn is a decent wellspring of flavonoids activity of Hippophaerhamnoides might be as well, fundamentally quercetin, kaempferol, because of its higher phenolic and flavonoids myricetin, and isorhamnetin, and a significant substance 8. Hippophaerhamnoides gives a wellspring of tocopherols17.Phytochemical superb wellspring of basic unsaturated fats examination of the plant affirmed the that is 70% of its composition. The seed oil is nearness of mineral acids, carotenes, utilized as hostile to maturing, antioxidant, flavonoids, nutrients, sugars and amino acids anti-inflammatory and as regular UV blocking 18. Sea buckthorn is esteemed for its cancer operator in restorative plans. The inhibitory prevention agent, cardioprotective, impacts of alcoholic leaf and organic product antiatherogenic, antidiabetic, concentrate of Hippophaerhamnoides have hepatoprotective, hostile to cancer-causing, been examined to repress the oxidative harm immunomodulating, antiviral, antibacterial, instigated by chromium. Alcoholic leaf and mitigating and vasodilating impacts. Other natural product remove were found to significant properties of sea buckthorn forestall chromium initiated free extreme incorporate its cytoprotective impacts. creation and reestablished the antioxidants status. It has been indicated that product of PHARMACOLOGICAL ACTIVITIES OF Hippophaerhamnoides restrain nicotine instigated oxidative pressure. It has been Examination on restorative plants is exhibited by different in-vivo, in-vitro expanding step by step. contemplates that Hippophaerhamnoides has Hippophaerhamnoides is an enchantment antioxidants activity 10. plant as it contains a biodiversity of both dietary just as therapeutic constituents. This review discussed the pharmacological Antibacterial Activity activities of Hippophaerhamnoides. Antibacterial activity of Hippophaerhamnoides leaf removes

3 IJRPC 2021, 11(1), 1-9 Athar Jamal et al. ISSN: 22312781 alongside antibiotic medication utilized as subordinate way. Pretreatment with standard was assessed against Escherichia casuarinin diminished TNF- - actuated ace coli (MTCC 739), Arthobacterprotophormial provocative middle people, for example, IL-1, (MTCC 2682) and Micrococcus luteus IL-6, IL-8, and MP-1. Further, in the murine (MTCC 106). Affectability of various bacterial macrophage cell line, SBT leaf alcoholic strains to Hippophae was estimated as far as concentrate altogether repressed the zone of hindrance utilizing agar-well upgraded creation of NO prompted by LPS dispersion test strategy and paper plate (lipopolysaccharide) in a subordinate way dissemination technique. The and by its inhibitory impact on iNOS Hippophaerhamnoides methanolic leaf stimulation. As of late, SBT leaf alcoholic extract showed wide range against human concentrate have appeared directed antigen pathogens. The Hippophaerhamnoides introduction capacity of macrophages in methanolic leaf was discovered best against matured mice, which showed its resistant E. coli in which a most extreme zone of boosting and hostile to maturing impact. Sea restraint (24mm) was seen at a convergence buckthorn berries additionally indicated of 25 mg mL-1 while hindrance for (8mm) for immunoprotective impact against T-2 poison M. luteus2. Michel et al. 12gave detailed initiated immunodepression in 15-day-old antibacterial activity of chicks. SBT has been broadly utilized in Hippophaerhamnoides using distinctive oriental customary drugs for treatment of concentrate against Staphylococcus aureus. numerous incendiary issues. Henceforth from The current investigation demonstrated that these perceptions, the mitigating and 6mg/ml of both the n-hexane and chloroform immunomodulatory exercises have been concentrates of Hippophaerhamnoides deductively demonstrated 12. berries and 6mg/ml of n-hexane concentrate of leaves of the plant altogether (p<0.05) Anticancer Activity hindered the development of methicillin-safe In an investigation on MDA-MB-231 human Staphylococcus aureus. Antibacterial activity bosom malignancy cells, Sea buckthorn againstStaphylococcus aureus was appeared procyanidins disconnected from the seeds by the various concentrates that was non- have inhibitory effects on fatty acid synthase noteworthy (p>0.05). This antibacterial action (FAS): a key compound for all over again might be huge at higher dosages, that long-chain unsaturated fat biosynthesis, estimation is just conceivable by disengaging significant levels of which are found in the dynamic constituents. The n-hexane malignant growth cells. This restraint was concentrate of leaves of the plant have portion subordinate at fixations running from antibacterial action against methicillin-safe 0 to 0.14µg/ml. A grouping of 0.087µg/ml Staphylococcus aureus (MRSA) 14. repressed half of FAS action. In addition, cell development was smothered by treatment Anti-inflammatory and with sea buckthorn procyanidins at focuses immunomodulatory Activity somewhere in the range of 10 and 60µg/ml. The SBT leaf separate was found to have Likewise, the tried procyanidins were found critical calming action in adjuvant incited joint to initiate cell apoptosis in a portion inflammation (AIA) rodent model and subordinate way. The creators propose that lipopolysaccharide actuated provocative these procyanidins can actuate MDA-MB-231 reaction in murine macrophages 12. In cell apoptosis by hindering intracellular FAS another investigation, disengaged casuarinin action 7. from the SBT leaves was investigated for the Olsson et al. 21 looked at the effect of 10 impact on the TNF- -initiated ICAM-1(intra different concentrates of foods grown from cellular cell adhesion molecule) articulation in the ground, including sea buckthorn berries, a human keratinocytes cell line. Pre- on the multiplication of HT29 semi-colon treatment with casuarinin restrained TNF- - malignant growth cells and MCF-7 breast actuated protein and mRNA articulation of cancer cells. They saw that sea buckthorn ICAM-1 and ensuring monocyte had the most noteworthy hindrance effect for adhesiveness in HaCaT cells. Casuarinin the multiplication of HT29 and MCF-7 cells at altogether restrained TNF- - actuated its two most noteworthy directed focuses enactment of NF-B, ERK and p38 MAPK in a (0.25 and 0.5%). The creators propose that

4 IJRPC 2021, 11(1), 1-9 Athar Jamal et al. ISSN: 22312781 the hindrance of malignancy cell phenolic mixes, for example, flavonoids, multiplication was corresponded with including kaempferol, quercetin, and convergences of carotenoids and . isorhamnetin; these shield cells from Boivin et al.22 decided the antiproliferative oxidative harm that can prompt hereditary action of the juices of 13 sorts of berries, change and to malignancy 13. The current including ocean buckthorn, at centralizations examination was attempted to survey the of 10–50µg/ml against five disease cell lines counter tumor movement of isorhamnetin in vitro: AGS—stomach adenocarcinoma, secluded from H. rhamnoides L., utilizing ACF-7—mammary organ adenocarcinoma, hepatocellular carcinoma cells (BEL-7402). PC-3—prostatic adenocarcinoma, Caco2— We found that the cytotoxic impacts of colorectal adenocarcinoma, and MDA-MB- isorhamnetin demonstrated portion and time- 231—mammary organ adenocarcinoma. It reliance against BEL-7402 cells. We utilized was discovered that sea buckthorn berry the MTT [3-(4, 5-dimethylthiazol-2-yl)-2,5- juice, similar to blackberry and dark diphenyltetrazolium bromide] test to test the chokeberry juices, had antiproliferative cytotoxic impacts of isorhamnetin, and found properties. In any case, no connection was that isorhamnetin displayed powerful found between the counter proliferative cytotoxicity against BEL-7402 cells, with IC50 properties of the berry juices and their cancer equivalent to 74.4±1.13gml−1 for brooding prevention agent limit, and the hindrance of 72h. Specifically, we found that the greater malignant growth cell expansion by the juices part of the BEL-7402 cells were disconnected didn't include caspase-subordinate from the outside of culture dish and were apoptosis. In spite of this, concealment of drifting in the medium after treatment with tumor necrosis factor (TNF)-instigated generally high centralization of isorhamnetin. initiation of nuclear factor kappa-light-chain Moreover, cytotoxicity of the flavonols on enhancer of activated B cells (NFκB) was tumor cells relies upon cell collection of the examined. The anticancer activity of sea medications. The measure of isorhamnetin buckthorn organic products might be founded gathered in BEL-7402 was examined by on its improvement of the DNA- binding high-performance liquid chromatography action of interferon regulatory factor-1 (IRF- (HPLC) and indicated that isorhamnetin 1), a known antioncogenic interpretation could saturate the cell film into the cell. The factor causing development concealment and estimation of isorhamnetin amassed in cells apoptosis. is little 23. Zhang et al. 24 exhibited that infusion of seed oil (1.59 g/kg body weight) Ant diabetic Activity significantly repressed the development pace Ant diabetic movement was estimated by of transplanted melanoma (B16) and alpha-glucosidase inhibitor measure at sarcoma (S180) tumors in mice. various centralizations of the example running from 0.0625 to 0.50 mg/ml. Fluid leaf Wound-Healing Activity separate demonstrated low alpha The defensive and therapeutic impacts of glucosidase hindrance movement, that is, SBT against wounds, consumes, singes, under half even at the most elevated fixation ulcers and mucosal wounds have been (0.50 mg/ml) taken in the investigation and broadly examined utilizing distinctive creature hence IC50 value couldn't be resolved. MeOH models and by clinical preliminaries. In the leaf separate showed better antidiabetic ongoing years, SBT leaf extricate has been action with 73.67% compound hindrance at experimentally explored and indicated that it 0.50 mg/ml fixation and IC50 esteem was improves intense and constant dermal injury seen as 0.25 mg/ml. This mirrored mending (consumes and diabetic) in rodents. methanolic concentrate of sea buckthorn The SBT rewarded creature demonstrated leaves contains certain compound(s) with quicker decrease in twisted region in alpha glucosidase inhibitory movement 19. correlation with control and silver sulfadiazine (standard consideration) rewarded creatures. The effective use of SBT expanded Anti-tumor Activity neovascularization, collagen blend and The antitumor activity of sea buckthorn can adjustment at twisted site, as prove by up- be ascribed to antioxidant mixes, especially controlled articulation of vascular endothelial

5 IJRPC 2021, 11(1), 1-9 Athar Jamal et al. ISSN: 22312781 development factor, collagen type-III, network metalloproteinases (MMP-2, 9) and Anti-stress and adaptogenic Activity expanded substance of hydroxyproline and The natural details which can improve hexosamine. SBT treatment likewise physical perseverance, mental capacity and expanded the endogenous enzymatic and vague opposition of the body to pressure nonenzymatic antioxidant and diminishing in have been named as adaptogen. The lipid peroxide levels in wound granulation adaptogenic movement of SBT leaf tissue. Further, it has been accounted for that extricates was concentrated in rodents SBT leaf has no cytotoxic, overwhelming utilizing a latent cold (5 ℃) hypoxia (428 metal defilement and antagonistic impact mmHg) restraint (CHR) creature model. SBT after oral organization. Flavonoids are one of leaf fluid concentrate controlled orally in the significant naturally dynamic substances rodents at a portion of 100 mg/kg BW both in present in SBT leaves and organic products. single and five dosages 30 min before CHR The positive mending impact of effective introduction had huge enemy of stress and utilization of SBT flavone (segregated from adaptogenic action. The impact of the organic product mash) (1.0%, w/v) has been concentrate was concentrated on lipid seen on dermal injuries in test rodents 12. peroxidation and cancer prevention agent boundaries in liver and gastrocnemius Hepatoprotective Activity muscle of rodents. Results proposed that The liver is regularly influenced by a large supplementation with SBT leaf extract number of natural contaminations and decreased the oxidative worry in liver and medications, all of which place a weight on muscle of rodents during CHR presentation this crucial organ that can harm and and post-stress recuperation. During serious debilitate the liver and in the end lead to unpleasant introduction to CHR and post hepatitis or cirrhosis. The hepatoprotective pressure recuperation the vigorous digestion action of SBT leaves and seed oil was just as hexose monophosphate pathway is assessed utilizing carbon tetrachloride (CCl4) stifled. The single and five dosages of SBT instigated hepatic harm in creatures, and the separate treatment confined the abatement outcomes indicated that both SBT leaf or better kept up tissue glycogen and alcoholic concentrate and seed oil enhanced compound exercises for example CCl4-actuated liver injury as confirm by both hexokinase, phosphofructokinase, citrate histological and biochemical discoveries. In synthase and glucose-6-phosphate an ongoing report by Maheshwari et al. 25, a dehydrogenase, in blood, liver and muscle, portion of the phenolic constituents of leaves, during CHR introduction. The examinations for example, gallicacid, myricetin, quercetin, recommended that SBT leaf fluid concentrate kaempferol and isorhamnetin were treatment made a pattern for moving distinguished in the phenol rich part (PRF) by anaerobic digestion oxygen consuming reverse phase high performance liquid during different pressure CHR presentation chromatography (RP-HPLC). Oral and post pressure recuperation 12. organization of PRF at portion of 25–75 mg/kg BW fundamentally shielded from CCl4 Cardio protective and anti-atherogenic initiated height in aspartate Activity aminotransferase, alanine aminotransferase, Healing impact of SBT arrangements on - glutamyltranspeptidase and bilirubin in cardiovascular illnesses are known and all serum, upgraded the hepatic antioxidants. around reported in Tibetan customary clinical These perceptions propose that PRF has writing. A few investigations on human and intense antioxidantaction and ensured creatures have been completed to assess against CCl4 prompted oxidative harm in the the impact of flavonoids of SBT on liver. Be that as it may, increasingly precise cardiovascular sicknesses, as some investigations on interminable liver injury mixes are known to have positive ought to be done for the advancement of an ionotropic impacts. Flavonoids in SBT foods item or a plan. SBT might be a cheerful grown from the ground are notable to medication for avoidance and treatment of improve the working of cardiovascular liver fibrosis, yet further all around controlled framework. The total flavonoids of Hippophae clinical preliminaries are required. (THF), extricated from its leaves and organic

6 IJRPC 2021, 11(1), 1-9 Athar Jamal et al. ISSN: 22312781 product, is a gathering of mixes containing blood-determined human macrophages with seven sorts of flavonoids. Among them, the a reduction and increment in TNF-and IFN- principle segments are isorhamnetin and separately12. quercetin. THF treatment displayed defensive impacts on myocardial ischemia and reperfusion, tumors, oxidative injury and maturing 12. The investigation by Pang et al. 26 exhibited the antihypertensive impact of absolute flavones extricated from seed buildups of SBT in ceaseless sucrose took care of rodents by controlling its insulin and angiotensin II levels. The hypertension, hyperinsulinemia, dyslipidemia and actuated angiotensin II incited by the high-sucrose diet could be improved by absolute SBT flavones and the ideal impact was seen at the portion of 150 mg/kg/day. SBT supercritical CO2- removed seed oil have been appeared to have hostile to atherogenic and Fig. 1: Berries of H. rhamnoides cardioprotective exercises. In an investigation utilizing bunny as a creature model, organization of SBT seed oil alongside elevated cholesterol diet confined further ascent of absolute cholesterol and caused a noteworthy decay of triglyceride and (low density lipoprotein) LDL-cholesterol when contrasted with creatures benefited from elevated cholesterol diet as it were. The ascent in (high density lipoprotein) HDL- cholesterol over the basal qualities in seed oil rewarded creatures was altogether higher than the non-rewarded creatures. SBT seed oil additionally caused a huge vasorelaxation Fig. 2: Leaves of H. rhamnoides 27.

Anti-viral and antimicrobial Activity CONCLUSION A precise substance examination of dynamic Present study provides review about divisions from the SBT leaves has prompted medicinal importance of the revelation of another phytochemical Hippophaerhamnoides. All plant parts are medicate Hiporamin, having a wide range of useful in treating different diseases and antiviral and antimicrobial exercises. ailments very effectively. Recent review also Hiporamin is a filtered portion of polyphenol reveals many pharmacological activities of division, containing monomeric hydrolysable Hippophaerhamnoides like antibacterial, gallo-ellagi-tannins. It was found to have a antiviral, adaptogenic, antistress, anti solid enemy of infection action and wide atherogenic, cardio protective, wound scope of activity against Influenza and healing, hepatoprotective, anti-tumor, anti- Herpes infections. Antiviral movement of diabetic, anti-cancer anti-inflammatory Hiporamin in regard of Influenza infection activities. was seen by its inhibitory impact on the viral neuraminidase. It likewise demonstrated inhibitory impact in a HIV contamination in the cell culture and antimicrobial movement. The SBT leaf extract additionally has a noteworthy enemy of dengue action when REFERENCES assessed in Dengue infection type-2 tainted

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