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Background: Chlorophytum borivilianum L. is a recognized herbal medicine for the management of impotency in South Asian countries. In Ayurveda, it is used for the management of multiple health conditions, including diabetes, infection, and cardiovascular diseases. Parts of the plant have been used as excellent antioxidants and scavengers of free radicals. Since oxidative stress plays an important role in spermatogenesis and fertility in male populations, this study evaluated the role of ethanolic extract of C. borivilianum roots in epididymal sperm maturation against adversities posed by ionizing gamma irradiation. Materials and methods: Antioxidant potential of C. borivilianum root extract (CRE) was evaluated through DPPH (2,2-diphenylpicrylhydrazyl) and NO (nitric oxide) scavenging assays. Four groups of healthy Swiss albino mice were constituted, which were labeled as follows: Group I: sham control, Group II: 7-day pre-treatment with 50 mg/kg CRE, Group III: 6 Gy irradiation without pre-treatment, and Group IV: 7-day pre-treatment with 50 mg/kg CRE and 6 Gy irradiation on day 7. Swiss albino mice were observed for 30 days and later sacrificed to evaluate sperm quality parameters. Results: CRE showed a remarkable antioxidant potential with IC50 values of 46.37 µg/ml and 98.39 µg/ml for DPPH and NO, respectively. A significant decline (p < 0.001) in cauda epididymal sperm count, motility, and viability was observed in Group III animals. Group IV also showed a substantial decline (p < 0.01) in all three parameters compared to Group I; nonetheless, these were significantly higher than Group III. Morphological alterations indicated a coiled and bent tail, with the presence of cytoplasmic droplets in Group III, which declined substantially in Group IV. The ultrastructure of sperm indicated higher curvature of hook in Group III than Group IV, indicating specific interferences in the sperm maturation process. Conclusion: It was concluded that pre-treatment with 50 mg/kg body weight of CRE could protect sperm during epididymal maturation against oxidative stress.

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KEYMESSAGE: CbSE overexpression increased stigmasterol levels and altered plant morphology. The genes upstream and downstream of CbSE were found to be upregulated, which confirms its regulatory role in the saponin biosynthetic pathway. Chlorophytum borivilianum is a high-value medicinal plant with many promising preclinical applications that include saponins as a major active ingredient. Squalene epoxidase (SE) is one of the major rate-limiting enzymes of the saponin biosynthetic pathway. Here, we functionally characterized C. borivilianum SE (CbSE) by over-expressing heterologously in Nicotiana tabacum. The heterologous expression of CbSE resulted in stunted pant growth with altered leaf and flower morphology. Next, RT-qPCR analysis of transgenic plants overexpressing CbSE revealed increased expression levels of Cycloartenol synthase (CAS), Beta amyrin synthase (ßAS), and cytochrome P450 monooxygenase 51 (CYP51) (Cytochrome P450), which encode key enzymes for triterpenoid and phytosterol biosynthesis in C. borivilianum. Further, Methyl Jasmonate (MeJa) treatment upregulated Squalene synthase (SQS), SE, and Oxidosqualene cyclases (OSCs) to a significant level. GC-MS analysis of the leaf and hairy roots of the transformants showed an increased stigmasterol content (0.5-1.0 fold) compared to wild type (WT) plants. These results indicate that CbSE is a rate-limiting gene, which encodes an efficient enzyme responsible for phytosterol and triterpenoid production in C. borivilianum.

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Eighteen crossbred Karan Fries (KF) cows in mid-lactation (av. 130 days) were selected from the livestock herd of the institute. The treatment for the experimental cows was as follows: no supplement (control), a low dose of Chlorophytum borivilianum (CB) at a dose rate of 40 mg/kg BW/day (T1, n = 6), and a high dose of CB at a dose rate of 80 mg/kg BW/day (T2, n = 6) for a period of 90 days in hot-humid season. Physiological responses like respiration rate (RR), pulse rate (PR), and rectal temperature (RT) were recorded in the morning (8.00 A.M.) and afternoon (2.30 P.M.) at weekly intervals. The expression of pro-inflammatory (IL-1ß and TNF-α) and anti-inflammatory cytokine (IL-10) in PBMCs, the plasma level of antioxidants (SOD, catalase, TBARS, and TAC), hormones (cortisol, prolactin), and energy metabolites (glucose, NEFA, urea, and creatinine) were determined. Dry and wet bulb temperatures and minimum and maximum temperatures were recorded, and the THI was calculated. The values of PR and RT were decreased (p < 0.01) in the T2 group in comparison to T1. Plasma glucose level was lower, and NEFA, urea, and creatinine level were higher (p < 0.01) in the control group as compared to T2 and T1 groups, respectively. Plasma cortisol and prolactin levels were significantly higher (p < 0.01) in the control group and were lower in T1 and T2 groups. Feeding of CB in high dose decreased (p < 0.01) plasma SOD, catalase, TBARS, and improved TAC levels in T2 over the T1 group. The dietary supplementation of CB at a dose rate of 80 mg/kg BW/day was more effective in lowering the stress level and augments the immunity by downregulating pro-inflammatory and anti-inflammatory cytokine levels. Therefore, dietary CB supplementation could be used as an effective heat stress ameliorator in dairy cows.

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BACKGROUND: Radiation therapy is considered as an important tool in cancer treatment. Despite its impressive role in treating cancer, severe side effects in organs have been reported. To address these therapeutic side effects, several combination methods have been identified to minimize adverse effects caused by radiation therapy. AIMS AND OBJECTIVES: Based on higher radioactive sensitivity of testicular tissues, administration of Chlorophytum borivilianum (CB) Sant. F extracts was evaluated for its protective effects against radiation in testis. MATERIALS AND METHODS: Two forms of CB extracts (CB alone and CB-silver nanoparticles [AgNPs]) were administered at a dose of 50 mg/kg body weight in Swiss albino male mice for 7 consecutive days. Following 6 Gy gamma radiation, animals were observed for 30 days in four phases. Sperm counts, body weight, testicular weight and stereological and histological evaluation of testis were evaluated. RESULTS: Following irradiation, a significant decline in body weight (P = 0.008) and testicular weight (P = 0.001) was noted when compared with control. Ununiformed type A and B spermatogonia, partially filled tubules, inter-tubular vacuoles, and disrupted epithelium were the main types of damages caused by irradiation. Reorganization and resumption of histological features emerged from the 15th day postirradiation in CB extract (CBE)-treated animals. CONCLUSION: Testicular response was observed against radiation in animals treated with CB extracts, while CB-AgNPs indicated better toleration when compared to CB extract alone.

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Plant specific miRNAs (Novel miRNAs) are well known to perform distinctive functions in biological processes. Identification of new miRNAs is necessary to understand their gene regulation. Degradome provides an opportunity to explore the miRNA functions by comparing the miRNA population and their degraded products. In the present study, Small RNA sequencing data was used to identify novel miRNAs. Further, degradome sequencing was carried out to identify miRNAs targets in the plant, Chlorophytum borivilianum. The present study supplemented 40 more novel miRNAs correlating degradome data with smallRNAome. Novel miRNAs, complementary to mRNA partial sequences obtained from degradome sequencing were actually targeting the later. A big pool of miRNA was established by using Oryza sativa, Arabidopsis thaliana, Populus trichocarpa, Ricinus communis, and Vitis vinifera genomic data. Targets were identified for novel miRNAs and total 109 targets were predicted. BLAST2GO analysis elaborate about localization of novel miRNAs' targets and their corresponding KEGG (Kyoto Encyclopedia for Genes and Genomes) pathways. Identified targets were annotated and were found to be involved in significant biological processes like Nitrogen metabolism, Pyruvate metabolism, Citrate cycle (TCA cycle), photosynthesis, and Glycolysis/Gluconeogenesis. The present study provides an overall view of the miRNA regulation in multiple metabolic pathways that are involved in plant growth, pathogen resistance and secondary metabolism of C. borivilianum.

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BACKGROUND: MicroRNAs act as molecular regulator of cell signaling, plant growth and development, and regulate various primary and secondary plant metabolic processes. In the present study, deep sequencing of small RNAs was carried out to identify known and novel miRNAs from pharmaceutically important plant, Chlorophytum borivilianum. RESULTS: Total 442 known miRNAs and 5 novel miRNAs were identified from young leaf small RNA library. Experimental validation with stem loop RT-PCR confirmed the in silico identification. Based on transcriptome data of root and leaf of C. borivilianum, Oryza sativa, and Arabidopsis thaliana target gene prediction was done using psRNAtarget and mirRanda. BLAST2GO helped in localization of predicted targets and KEGG (Kyoto Encyclopedia for Genes and Genomes) pathway analysis concluded that miR9662, miR894, miR172, and miR166 might be involved in regulating saponin biosynthetic pathway. The correlation between miRNA and its target gene was further validated by RT-qPCR analysis. CONCLUSION: This study provides first elaborated glimpse of miRNA pool of C. borivilianum, which can help to understand the miRNA dependent regulation of saponin biosynthesis and to design further metabolic engineering experiment to enhance their contents in the plant.

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Chlorophytum borivilianum is an important species of liliaceae family, owing to its vital medicinal properties. Plant roots are used for aphrodisiac, adaptogen, anti-aging, health-restorative and health-promoting purposes. Saponins, are considered to be the principal bioactive components responsible for the wide variety of pharmacological properties of this plant. In the present study, we have performed de novo root transcriptome sequencing of C. borivilianum using Illumina Hiseq 2000 platform, to gain molecular insight into saponins biosynthesis. A total of 33,963,356 high-quality reads were obtained after quality filtration. Sequences were assembled using various programs which generated 97,344 transcripts with a size range of 100-5,216 bp and N50 value of 342. Data was analyzed against non-redundant proteins, gene ontology (GO), and enzyme commission (EC) databases. All the genes involved in saponins biosynthesis along with five full-length genes namely farnesyl pyrophosphate synthase, cycloartenol synthase, ß-amyrin synthase, cytochrome p450, and sterol-3-glucosyltransferase were identified. Read per exon kilobase per million (RPKM)-based comparative expression profiling was done to study the differential regulation of the genes. In silico expression analysis of seven selected genes of saponin biosynthetic pathway was validated by qRT-PCR.

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Background Chlorophytum borivilianum is a rare medicinal plant originally distributed throughout the forest of India. The tubers of C. borivilianum are used as an aphrodisiac and impotence supplement. The propagation of C. borivilianum is possible through seeds and tubers, but conventional methods may take several months. Hence in vitro technique of shoot regeneration could be an efficient alternative means of propagating the species. Latest study reported microtuberization of C. borivilianum but there is no sufficient study on a rapid method for shoot multiplication and elongation. Results Young shoot buds of C. borivilianum were cultured on MS medium containing 6-benzylaminopurine (BAP) and Kinetin (Kn), both at 0, 8.88, 17.8 and 26.6 µM, either individually or in combinations. Proliferated shoots were subcultured on fresh medium of the same constituents on week 3 of culture for further shoot multiplication and elongation. BAP alone (8.88-26.6 µM) was significantly effective on shoot multiplication, while Kn alone (8.88-26.6 µM) was significantly effective on shoot elongation compared to the control containing MS basal medium without any plant growth regulator. However, combination of both cytokinins stimulated an interaction producing higher shoot number and shoot length compared to their individual application. Conclusions The most suitable combination was 8.88 µM BAP + 8.88 µM Kn, reaching a mean shoot number of 10.83 and shoot length of 6.85 cm.

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UNLABELLED: The effect of C. borivilianum root on blood glucose, glycated hemoglobin (HbAIc), insulin and lipid profile levels in diabetes mellitus are not fully understood. This study therefore investigated the effect of C. borivilianum root on the above parameters and oxidative stress of the pancreas in diabetes. METHODS: C. borivilianum root aqueous extract (250 and 500 mg/kg/day) was administered to streptozotocin (STZ)-induced male diabetic rats for 28 days. Body weight, blood glucose, HbA1c, insulin, lipid profile levels and glucose homeostasis indices were determined. Histopathological changes and oxidative stress parameters i.e. lipid peroxidation (LPO) and antioxidant enzymes activity levels of the pancreas were investigated. RESULTS: C. borivilianum root extract treatment to diabetic rats maintained near normal body weight, blood glucose, HbA1c, lipid profile and insulin levels with higher HOMA-ß cell functioning index, number of Islets/pancreas, number of ß-cells/Islets however with lower HOMA-insulin resistance (IR) index as compared to non-treated diabetic rats. Negative correlations between serum insulin and blood glucose, HbA1c, triglyceride (TG) and total cholesterol (TC) levels were observed. C. borivilianum root extract administration prevented the increase in lipid peroxidation and the decrease in activity levels of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) with mild histopathological changes in the pancreas of diabetic rats. CONCLUSIONS: C. borivilianum root maintains near normal levels of these metabolites and prevented oxidative stress-induced damage to the pancreas in diabetes.

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Increase in male sexual dysfunction, and its treatment with conventional aphrodisiac drugs with side effects lead to investigate the spermatogenesis and androgenesis augmentative efficacy of hydromethanolic (40 : 60) extract of root of Chlorophytum borivilianum (family - Liliaceae) against cyproterone acetate-induced subfertility in Wistar strain male albino rat. For this purpose, experimental rats were divided into three treatment groups: vehicle (received distilled water), cyproterone acetate (gastric intubation at 250 mg kg(-1) twice daily for 35 days) and cyproterone acetate plus root extract of C. borivilianum (gastric intubation at 250 mg kg(-1) plus 400 mg kg(-1) with an interval of 20 min twice daily for 35 days). After 35-day treatment, all rats were euthanised. Reproductive deviations towards negative side were investigated by screening the spermatogenic and steroidogenic biosensors. Oxidative stress profile in reproductive organs and sperm pellet was evaluated by biochemical assessment of antioxidative enzyme activities and level of end products of the lipid peroxidation. Apoptosis profile was evaluated by Western blot study, TUNEL assay and DNA fragmentation study of testicular tissues. Evaluation of toxicity profile was included for experimental investigation. After cyproterone acetate treatment, the pituitary-testicular axis was deviated towards the negative side and its tuning system was affected by oxidative stress and apoptosis-mediated process, which reduced the quality of semen and finally led to subfertility. Co-administration of C. borivilianum root extract enhanced male reproductive potentiality and prevented the negative deviations after the treatment with cyproterone acetate by means of increasing oxidative defence and maintaining homeostasis in testicular apoptosis process.

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ETHNOPHARMACOLOGICAL RELEVANCE: Safed musli (Chlorophytum borivilianum) is an eminent medicinal plant of India and considered as a 'white gold' or 'divya aushad' in Indian systems of medicine. In Ayurveda, Chlorophytum borivilianum belongs to the group of "Vajikaran Rasayana" corroborated to its rejuvenating, aphrodisiac, natural sex tonic properties and effective in alleviating sexual disorders. It is largely used as ethnic medicine by local healers of indigenous communities of India. MATERIALS AND METHODS: A thorough bibliographic investigation was carried out by analyzing worldwide accepted scientific data base (Pub Med, SciFinder, Scopus and Web of Science), thesis, recognized books, non impact and non indexed journals. RESULTS: Traditionally, Chlorophytum borivilianum is well known for treating male impotency in India. The multi therapeutic and nutritional importance of Chlorophytum borivilianum is attributed to the rich source of phytochemicals particularly saponins. Recently, Chlorophytum borivilianum has gained a well established domestic (Indian) and international market for being the herbal alternative of "Viagra" without any side effects. Under the trade name 'Nai Chetna', the state government of Gujarat, India, has launched a novel potency drug from Chlorophytum borivilianum. Modern pharmacological studies of Chlorophytum borivilianum have demonstrated a wide range of pharmacological activities, most importantly aphrodisiac, immunomodulatory and anticancer activities. CONCLUSION: The increased commercial exploitation of Chlorophytum borivilianum and low productivity of this endangered plant has raised the concern over its conservation. It has been envisaged that efforts should be made to standardize, encourage and popularize the cultivation of Chlorophytum borivilianum as a commercial crop. The analysis of previous pharmacological investigations suggested lack of substantial scientific evidences in various studies and do not stand the test of critical assessment. Due to high economic value, Chlorophytum borivilianum has also encountered a problem of adulteration with closely resembling medicinally inferior species. The studies available on toxicity, safety and quality of Chlorophytum borivilianum are inadequate for providing information on commercial utilization. Thus, the present review summarizes comprehensive information on Chlorophytum borivilianum and possible scope for future research to fill the existing lacunae on its different aspects of the study.

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This study focuses on the establishment of in vitro tuberization of Chlorophytum borivilianum using solid and liquid culture systems. A high in vitro tuberization rate on solid and stationary liquid Murashige and Skoog media was observed in the presence of 60 g l⁻¹ sucrose with 950, 1,265 and 1,580 µM 2-chloroethyl-trimethylammonium chloride (CCC). Application of a higher sucrose concentration of 90 g l⁻¹ showed a negative interaction with CCC on in vitro tuber number and days to in vitro tuber induction. For economic feasibility, 950 µM CCC with 60 g l⁻¹ sucrose was chosen as the best combination for in vitro tuberization in both solid and stationary liquid media. For optimization of in vitro tuber production,a comparison between solid, stationary liquid and shake liquid culture was carried out. Liquid culture with shaking at 80 r.p.m. resulted in a >2.5-fold increase in in vitro tuber production compared with solid culture.

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Shweta Musali (Chlorophytum borivilianum (CB)) is a traditionally used herb for its benefits in male sexual and general health. In the recent past, the herb has attained much commercial significance, both in domestic and international markets. However, limited clinical data is available to establish its traditional claims. The present study was aimed at evaluating the effect of the water soluble extract of CB root tubers on semen and testosterone in healthy adult males. The research was designed as a randomized, double-blind, placebo-controlled, trial upon the volunteers registered from the outpatient department (OPD) with age ranging from 20 to 40 years. Water extracts of CB and placebo was administered in the patients of groups A and B, for 12 weeks, in two divided doses of 500 mg. Assessment was done based upon Semen (Volume, Liquefaction Time, Sperm Count, Sperm motility) and Serum Testosterone levels parameters. Highly significant improvement was noted in the above parameters after administration of CB extract in comparison to Placebo. Hence it was concluded that the trial drug was effective in improving male sexual health.

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BACKGROUND: Impaired sleep quality is commonplace within industrialized societies, as evidenced by the increasing number of prescription sleep aids available. Certain herbal preparations have been suggested to provide a natural benefit to sleep; however, limited controlled data are available documenting this benefit. In the present study we tested the effect of an experimental dietary supplement, containing the active ingredients Chlorophytum borivilianum and Velvet bean, on sleep quality using the Pittsburgh Sleep Quality Index (PSQI). METHODS: Eighteen healthy and active men and women, with evidence of impaired sleep quality, consumed the supplement daily for 28 days. The PSQI was administered before and after the intervention period. As indicators of safety, resting heart rate and blood pressure were measured, and a complete blood count, comprehensive metabolic panel, and lipid panel were determined. RESULTS: Sleep quality was influenced by the supplement, as evidenced by an improvement in every category of the PSQI questionnaire (P < 0.05), with most category scores improving approximately 50% from pre to post intervention. No adverse outcomes were noted with use of the supplement, as indicated by no change in resting heart rate, blood pressure, or any bloodborne parameter. CONCLUSIONS: An investigational dietary supplement containing the active ingredients Chlorophytum borivilianum and Velvet bean improves sleep quality in men and women. Additional placebo controlled trials are needed to corroborate these findings in individuals with self-reported sleeping difficulty.

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Efficient in vitro propagation of medicinally important endangered plant C. borivilianum has been achieved through somatic embryogenesis. Solid embryogenic medium [Murashige and Skoog medium containing 1.79 mM NH4NO3, 10.72 mM KNO3, 1.13 µM 2,4-dichlorophenoxyacetic acid, 7.38 µM 2-isopentenyladenine and 0.76 mM proline] supplemented with polyethylene glycol and sucrose (3 % each), exhibited 1.88-fold increase in embryo maturation compared to embryogenic medium containing 3 % sucrose. Liquid embryogenic medium supported better somatic embryo production and maturation. Highest total (79) and mature (cotyledonary stage) somatic embryos (38) as well as highest germination (57.5 %) was observed at inoculum density of 0.4 g/40 ml of liquid medium. 5.86 pH level exhibited optimal growth, maturation and germination of somatic embryos. Random amplified polymorphic DNA (RAPD) analysis of C. borivilianum plants regenerated through somatic embryogenesis revealed that they were genetically similar to the mother plant. The protocol established in the present study can be used for rapid mass multiplication of C. borivilianum in bioreactor employing liquid medium.

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BACKGROUND: Several isolated ingredients have been proposed to increase growth hormone (GH) release, including Chlorophytum borivilianum and Velvet bean. A combination of these two ingredients has been packaged within an investigational dietary supplement. It was the purpose of the present investigation to determine the impact of acute ingestion of this supplement on circulating GH in healthy, exercise-trained men. METHODS: Fifteen men ingested the dietary supplement on two different days, separated by one week. Blood was collected from subjects before ingestion of the supplement and at 20, 40, 60, 80, 100, and 120 minutes post ingestion. GH was analyzed in serum samples using an ELISA method. Values for GH for each subject, at each collection time, were averaged over both test days and used in the main analysis. RESULTS: Serum GH increased over time, with higher values at 60 minutes (1.56 ± 0.65 ng · mL(-1); P = 0.04; +767%), 80 minutes (1.76 ± 0.69 ng · mL(-1); P = 0.02; +878%), and 100 minutes (1.48 ± 0.62 ng · mL(-1); P = 0.05; +722%) compared to pre ingestion (0.18 ± 0.04 ng · mL(-1)). A great deal of subject variability existed in the area under the curve (AUC) for GH, with pooled values ranging from 0.49 to 61.2 ng · mL(-1) · 2 hr(-1.) CONCLUSION: Acute ingestion of an investigational dietary supplement containing the active ingredients Chlorophytum borivilianum and Velvet bean results in an increase in circulating GH in exercise-trained men. Additional placebo controlled investigations are needed to extend these findings. Moreover, studies are needed to determine if chronic use of such supplementation leads to favorable changes in health-related parameters associated with increased circulating GH.

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BACKGROUND: The present communication deals with the identification and characterization of bioactive principles from the roots of Chlorophytum borivilianum. METHOD: Methanolic extract and its fractions were used to isolate different phytoconstituents. The structures of isolated compounds were characterized and elucidated with chemical and spectroscopic techniques such as Infra Red, Nuclear Mass Resonace and Mass spectroscopy experiments. Fatty acids were characterized by GC-MS analysis. RESULT: Three Fatty acids were isolated and confirmed. One sterol stigmasterol was isolated. One new saponin named as Chlorophytoside-I (3ß, 5α, 22R, 25R)-26-(ß-D-glucopyranosyloxy)-22-hydroxy-furostan-12-one-3 yl O-ß-D-galactopyranosyl (1-4) glucopyranoside was isolated. CONCLUSION: The roots of Chlorophytum borivilianum contain three important fatty acids, common sterol stigmasterol and one furostanol saponins.

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Chlorophytum borivilianum Santapau & Fernandes (Liliaceae) is a very popular herb in traditional Indian medicine and constitute a group of herbs used as 'Rasayan' or adaptogen. Ethanolic extract of the roots and its sapogenin were evaluated for their immunomodulatory activity. Effect of azathioprine-induced myelosuppresion and administration of extracts on hematological and serological parameters was determined. Administration of extracts greatly improved survival against Candida albicans infection. An increase in delayed-type hypersensitivity response (DTH), % neutrophil adhesion and in vivo phagocytosis by carbon clearance method was observed after treatment with extracts. Immunostimulant activity of ethanolic extract was more pronounced as compared to sapogenins. The results, thus justifies the traditional use of C. borivilianum as a rasayana drug.