Ancient Herbal Medicine Research Articles

Exploring the geographical variability of Asphaltum punjabianum (Shilajit) from India in elements and phytochemicals variations using GC–MS/MS, LC, ICP OES, and in-silico studies

Shilajit is an ancient herbal medicine from India with many uses. It is a viscous material prominently present in the Himalayan and Hindukush rocks of the Indian region. The formation due to plant decompostion with time. Herbs assosciated to the stones reveals about composition of Shilajit. Primary objective is to create GCMS/MS and LC-analysis techniques for separating the phytochemicals and complete profiling of volatile and non-volatile metabolites found in Shilajit. Accordingly, an attempt was undertaken to develop a method for standardisation and, subsequently, certification inorganic content and purity of shilajit in various indian regions S-01, S-02, S-03, S-04, and S-05. Thus, an effort was made to create a fundamental technique for standardization and, consequently, authentication of shilajit that was acquired from various geographical locations.Trace metal concentrations were measured using ICP OES in mg Kg−1 for each sample. These elements included As, Pb, Cd, Ag, Al, B, Ba, Cr, Cu, Fe, Mn, Ni, Sb, Se, Zn, P, Ca, K, Mg, and Na. ICP OES analysis observed the significant concentration of Ca and K, in S-01 (102631 and 26779), S-02 (13529.25 and 36866.35), S-03 (8170.84 and 37989.10), S-04 (9616.37 and 18843.77), and S-05 (4992.10 and 2120.64), appropriately. By measuring the DPPH radical scavenging activity were assessed for their antioxidant activities of the S-01, S-02, S-03, S-04, and S-05 samples: 12.316 ± 0.561 %, 13.144 ± 2.884 %, 15.105 ± 1.421 %, 19.683 ± 2.013 %, and 0.0724 ± 1.045 %, respectively. Concurrently, TPC studies on Shilajit found between 10 and 150 mg g−1 wt Similarly, shilajit’s TFC was analysed with geographically. TFC were found in mg QCE/g of wt of S-01 12.56, S-02 5.56, S-03.48, S-04 49.23, and S-05 5809 each respectively. The phytochemicals protocatechuic acid and hippuric were tested for in silico anti-inflammatory and antihyperglycemic activity of acid, which need to be explored in-vivo applications.

POTENTIAL OF DAYAK ONION SKIN (ELEUTHERINE PALMIFOLIA) EXTRACT AS AN ALTERNATIVE INDICATOR FOR ACID BASE TITRATION PRACTICUM

Synthetic indicators such as phenolphthalein are used during acid-base titrations to determine the end point of the titration. These indicators are not environmentally friendly and the price is high. To overcome this, it is necessary to study the potential of natural indicators available in the environment, one of which was Dayak onion skin which has not been utilized after the tubers have been taken as an anti-cancer herbal medicine. The aim of this research was to determine the potential of Dayak onion skin (Eleutherine Palmifolia) as a natural indicator in acid-base titrations. The sampling collection of Dayak onions was done randomly in the city of Palangka Raya, Central Kalimantan. The method used in this study employed experimental techniques in the chemistry laboratory of FKIP ULM with the following steps: 1) Maceration of smoothed Dayak onion skin with 96% ethanol for 1, 2 and 3 days 2) Qualitative test of anthocyanin, color test with buffer pH 1-14, 3) Absorption measurement with a UV-Vis spectrophotometer at a wavelength of 200 - 400 nm, 4) Application of the extract as an indicator in acid-base titrations. The results showed that the optimum maceration time was on day 3 with an absorption of 4,520 at a wavelength of 380 nm. The application on the HCl – NaOH titration with a titration end point volume of 9.11 mL showed a color change from yellow to straw yellow, while the comparison indicator PP had a volume change of 8.57 mL with a color change from colorless to pink. The CH3COOH – NaOH titration gave a result of 11.43 mL and the PP indicator was 11.01 mL. For the NaHCO3 – HCl titration, the end point volume of the titration was 9.93 mL with a straw yellow – yellow color change and the MO indicator comparison was 10.27 mL with an orange – red color change. From the results of this research, there was a significant color change at the end point of the titration, so it can be stated that Dayak onion skin extract had the potential to be an alternative indicator in acid-base titration practice so that it can be a solution in reducing chemical pollution in the environment.

Molecular Targets and Mechanisms of Hedyotis diffusa Willd. for Esophageal Adenocarcinoma Treatment Based on Network Pharmacology and Weighted Gene Co-expression Network Analysis.

Hedyotis diffusa Willd. (HDW) is a common anticancer herbal medicine in China, and its therapeutic effectiveness has been demonstrated in a range of cancer patients. There is no consensus about the therapeutic targets and molecular mechanisms of HDW, which contains many active ingredients. To clarify the mechanism of HDW for esophageal adenocarcinoma (EAC), we utilized network pharmacology and weighted gene co-expression network analysis methods (WGCNA). The gene modules that were linked with the clinical features of EAC were obtained through the WGCNA method. Then, the potential target genes were retrieved through the network pharmacology method in order to determine the targets of the active components. After enrichment analysis, a variety of signaling pathways with significant ratios of target genes were found, including regulation of trans-synaptic signaling, neuroactive ligand-receptor interaction and modulation of chemical synaptic transmission. By means of protein-protein interaction (PPI) network analysis, we have successfully identified the hub genes, which were AR, CNR1, GRIK1, MAPK10, MAPT, PGR and PIK3R1. Our study employed molecular docking simulations to evaluate the binding affinity of the active components with the hub gene. The identified active anticancer constituents in HDW are scopoletol, quercetin, ferulic acid, coumarin, and trans-4-methoxycinnamyl alcohol. Our findings shed light on the molecular underpinnings of HDW in the treatment of EAC and hold great promise for the identification of potential HDW compounds and biomarkers for EAC therapy.

Mechanism of Nicotiflorin in San-Ye-Qing rhizome for anti-inflammatory effect in ulcerative colitis

BackgroundThe incidence of ulcerative colitis (UC) is on the rise globally and the development of drugs targeting UC is urgent. Finding the target of action of natural products is important for drug discovery, elucidation of drug action mechanism, and disease mechanism. San-Ye-Qing (SYQ), is an ancient herbal medicine, but whether the powder of its rhizome has pharmacological effects against UC and its mechanism of action are not clear. PurposeTo evaluate the therapeutic effectiveness of rhizome powder of SYQ in treating UC, and conduct an isolation and characterization of the chemical constituents of the powder. Further, screen the most potent compounds among them and determine the potential mechanism for treating UC. MethodsIn vivo, the therapeutic effect of SYQ's rhizome powder on UC was assessed by mice's body weight, DAI score, colon length, tissue MPO activity, serum inflammatory markers, etc. Additionally, HPLC was used to isolate and identify the specific chemical components of SYQ's rhizome powder. Then, the most effective compounds and their therapeutic targets were analysed and screened in SYQ rhizome powder using network pharmacology, combined with CCK-8 assay, NO release assay and molecular docking assay, in conjunction with CETSA, DARTS, SPR and enzyme activity assay. Finally, the biological effects of the key compound on the targets were validated using Western blot and ELISA. ResultsIn vivo, SYQ rhizome powder effectively restored mice's body weight, lowered DAI and pathological score, downregulated the expression of inflammatory biomarkers, and restored colon length, as well as the colonic epithelial and mucus barriers. Afterward, 9 compounds were isolated and identified from the powder of the rhizomes of SYQ by HPLC. Nicotiflorin is the primary compound in SYQ with the highest concentration. According to both CCK-8 and NO release tests, Nicotiflorin is also the most efficacious compound. Combined with network pharmacological prediction, molecular docking analysis, CETSA, DARTS, SPR and enzyme activity assay, Nicotiflorin may ultimately suppress inflammation by targeting p65 and inhibiting the NF-κB pathway, thereby attenuating the activation of NLRP3 inflammasome. To verify this conclusion, Western blot and ELISA experiments were conducted. ConclusionsOur results suggest that the extract from SYQ rhizomes has therapeutic properties for UC. Its active ingredient Nicotiflorin exerted potent anti-UC effects by binding to p65 and inhibiting the activation of NF-κB and NLRP3 inflammasomes.

Unlocking the potential of Lion’s Mane Mushroom (Hericium erinaceus)

The Lion's Mane mushroom, botanically known as Hericium erinaceus, stands out as a unique and esteemed member of the fungal kingdom. This extraordinary mushroom not only possesses an alluring appearance but also holds a significant historical presence in diverse cultures, especially within the context of ancient herbal medicine practices. This fungus holds promising prospects in several domains. Its potential as a natural remedy for cognitive health is gaining attention. This mushroom has neuroprotective properties and could play a role in supporting brain function, which is particularly relevant in the present aging population where neurodegenerative conditions like Alzheimer's disease are a growing concern. Furthermore, Lion's Mane has been explored for its potential in addressing mood disorders. It is a rich source of bioactive compounds, including β-glucans, that can positively affect the immune system. The fungus produces bioactive compounds that can be used to treat various chronic diseases like obesity, high blood pressure, hepatic disorders, and cancer ; it also has other benefits like wound healing and improving the immune system. This review endeavours to elucidate the multifaceted potential of Lion's Mane mushroom within the domains of nutrition, health, and wellness. Through a comprehensive examination of its properties and benefits, the review explored how Lion's Mane mushrooms can be harnessed to enhance human well-being. By unlocking the secrets hidden within this remarkable fungus, the study provides insights that can empower individuals to incorporate Lion's Mane into their daily lives, fostering a healthier and more balanced lifestyle.

Comprehensive Characterization of Triterpene Saponins in Rhizoma Panacis Japonici by Offline Two-Dimensional Liquid Chromatography Coupled to Quadrupole Time-of-Flight Mass Spectrometry.

Rhizoma Panacis Japonici (RPJ) is an ancient herbal medicine from China that has long been employed for its medicinal benefits in relieving arthritis physical debility and diverse afflictions. The primary bioactive constituents found in RPJ are triterpene saponins, which exhibit numerous pharmacological actions, including anti-inflammatory, antioxidant, and immunomodulating effects. The present study established a straightforward and effective approach for characterizing triterpene saponins in RPJ. An offline HILIC × RP LC/QTOF-MS method was developed, along with a self-constructed in-house database containing 612 saponins reported in the Panax genus and 228 predicted metabolites. The approach achieved good chromatographic performance in isolating triterpene saponins of RPJ, with the HILIC column as the first dimension (1D) and the BEH C18 column as the second dimension (2D). The developed two-dimensional liquid chromatography system exhibited an orthogonality of 0.61 and a peak capacity of 1249. Detection was performed using a QTOF mass spectrometer in a data-independent manner (MSE) in a negative ion mode. Using the in-house database, the collected MS data were processed by an automatic workflow on UNIFI 1.8.2 software, which included data correction, matching of precursor and product ions, and peak annotation. In this study, 307 saponins were characterized from RPJ and 76 saponins were identified for the first time in Panax japonicus. This research not only enhances our understanding of the chemical characteristics of RPJ but also offers a simple and efficient method for analyzing the complex composition of herbal medicine.

Essential oil from Ligusticum chuanxiong Hort. Alleviates lipopolysaccharide-induced neuroinflammation: Integrating network pharmacology and molecular mechanism evaluation

Ethnopharmacological relevanceChuanxiong, the rhizome of Ligusticum chuanxiong Hort., is an ancient herbal medicine that has gained extensive popularity in alleviating migraines with satisfying therapeutic effects in China. As the major bioactive component of Chuanxiong, the essential oil also exerts a marked impact on the treatment of migraine. It is widely recognized that neuroinflammation contributes to migraine. However, it remains unknown whether Chuanxiong essential oil has anti-neuroinflammatory activity. Aim of the studyTo explore the anti-neuroinflammatory properties of Chuanxiong essential oil and its molecular mechanisms by network pharmacology analysis and in vitro experiments. Materials and methodsGas chromatography-mass spectrometry (GC-MS) was used to identify the chemical components of Chuanxiong essential oil. Public databases were used to predict possible targets, build the protein–protein interaction network (PPI), and perform Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Moreover, cytological experiments, nitric oxide assay, enzyme-link immunosorbent assay, western blotting, and immunofluorescence assay were adopted to prove the critical signaling pathway in lipopolysaccharide (LPS)-induced BV2 cells. ResultsThirty-six compounds were identified from Chuanxiong essential oil by GC-MS, and their corresponding putative targets were predicted. The network pharmacology study identified 232 candidate targets of Chuanxiong essential oil in anti-neuroinflammation. Furthermore, Chuanxiong essential oil was found to potentially affect the C-type lectin receptor, FoxO, and NF-κB signaling pathways according to the KEGG analysis. Experimentally, we verified that Chuanxiong essential oil could significantly reduce the overproduction of inflammatory mediators and pro-inflammatory factors via the NF-κB signaling pathway. ConclusionChuanxiong essential oil alleviates neuroinflammation through the NF-κB signaling pathway, which provides a theoretical foundation for a better understanding of the clinical application of Chuanxiong essential oil in migraine treatment.

Ephedrae herba: A comprehensive review of its traditional uses, phytochemistry, pharmacology, and toxicology.

Ephedrae herba (called Mahuang in China) is the dried herbaceous stem of Ephedra sinica Stapf, Ephedra intermedia Schrenk et C. A. Mey., and Ephedra equisetina Bge. Ephedrae herba has a long history of use as an herb, and it was originally recorded in Sheng Nong's herbal classic. Ephedrae herba has also been widely used as both medicine and food. In the clinic, Ephedrae herba is commonly used for treating colds, bronchial asthma, nasal congestion, and other diseases. This review aims to provide a systematic summary on the traditional use, chemical constituents, pharmacological effects, clinical applications, quality control, toxicology, and pharmacokinetics of Ephedrae herba to provide a theoretical basis for further reasonable development of Ephedrae herba in clinical practice and creation of new drugs. Information on Ephedrae herba was gathered from various sources, including the scientific databases including CNKI, PubMed, SciFinder and ScienceDirect, classical books on traditional Chinese herbal medicine, Ph.D. and M.Sc. dissertations; Baidu Scholar; and from different professional websites. Ephedrae herba is distributed in regions of China and other areas. Ephedra and its compound preparations can be used for colds, bronchial asthma, nasal congestion and other diseases. Approximately 281 chemical constituents have been isolated from Ephedrae herba, including alkaloids, flavonoids, tannins, polysaccharides, volatile oils, organic acids, and other compounds. Among these constituents, alkaloids and volatile oils are the most abundant and represent the major bioactive constituents. Ephedrae herba possesses multiple pharmacological activities, including diuretic effect, anti-allergic effect, blood pressure regulatory, anti-inflammatory effect, anti-oxidation effect and anti-viral effects. Ephedrine hydrochloride and pseudoephedrine hydrochloride are generally selected as indicators for the quantitative determination of Ephedrae herba. The maximum dosage of Ephedrae herba should not exceed 10 grams. If overused, adverse reactions such as palpitations, sweating, irritability and insomnia will occur. Ephedrae herba is an ancient herbal medicine with a broad spectrum of pharmacological activities that has been used for thousands of years in China. It is one of the most commonly used herbal components of the TCM formulas. Hydrochloride and pseudoephedrine are the major bioactive constituents. However, there is a need to further understand the mechanisms of active components of Ephedrae herba. Future studies should perform an in-depth analyses of the pharmacokinetics and mechanisms of toxicity of Ephedrae herba. Quality standards should be developed to correspond to the various application methods to ensure the efficacy of drugs in actual treatment.

Platin-C containing nanoparticles: a recipe for the delivery of curcumin-cisplatin combination chemotherapeutics to mitochondria.

The success story of cisplatin spans over six decades now and yet it continues to be the key player in most chemotherapeutic regimens. Numerous efforts have been made to improve its efficacy, address its shortcomings, and overcome drug resistance. One such strategy is to develop new platinum(IV)-based prodrugs with functionally active ligands to deliver combination therapeutics. This strategy not only enables the drug candidate to access multiple drug targets but also enhances the kinetic inertness of platinum complexes and thereby ensures greater accumulation of active drugs at the target site. We report the synthesis of Platin-C, a platinum(IV)-based cisplatin prodrug tethered to the active component of ancient herbal medicine, curcumin, as one of the axial ligands. This combination complex showed improved chemotherapeutic efficacy in cisplatin resistant A2780/CP70 cell lines compared with the individual components. An amine-terminated biodegradable polymer was suitably functionalized with the triphenylphosphonium (TPP) cation to obtain a mitochondria-directed drug delivery platform. Quantification of Platin-C loading into these NPs using complementary techniques employing curcumin optical properties in high-performance liquid chromatography and platinum-based inductively coupled plasma mass spectrometry evidenced efficacious payload incorporation resulting in functional activities of both the components. Stability studies for a period of one week indicated that the NPs remain stable, enabling substantial loading and controlled release of the prodrug. The targeting nanoparticle (NP) platform was utilized to deliver Platin-C primarily in the mitochondrial network of cancer cells as monitored using confocal microscopy employing the green fluorescence of the curcumin pendant. Our studies showed that amine terminated NPs were relatively less efficient in their ability to target mitochondria despite being positively charged. This re-validated the importance of lipophilic positively charged TPP surface functionalities to successfully target cellular mitochondria. We validated the capabilities of Platin-C and its mitochondria-targeting nanoparticles towards inflicting mitochondria-directed activity in cisplatin-sensitive and cisplatin-resistant cell lines. Furthermore, our studies also demonstrated the effectiveness of Platin-C incorporated targeting NPs in attenuating cellular inflammatory markers by utilizing the curcumin component. This study advances our understanding of the cisplatin prodrug approach to combine chemotherapeutic and inflammatory effects in accessing combinatory pathways.

Herbal Medicines as Adjunctive Therapy for Oral Squamous Cell Carcinoma Management: a Case Report

Oral Squamous Cell Carcinoma (OSCC) is the most frequent oral cancer. The management for OSCC are surgery, chemotherapy, and radiotherapy. Chemotherapy and radiotherapy have high toxicity and side effects, therefore effort are needed to reduce them. Several studies show that herbal medicines have anticancer effects with lower toxicity and side effects. The objective is to discuss about management of OSCC using chemotherapy and anticancer herbal medicines as adjunctive therapy. A 59-year-old man came to the oral medicine clinic complaining a painful ulcer on the right tongue since 8 months ago. The ulcer had been enlarged in the last 3 months and the tongue was difficult to be moved. He used pain relievers and aloe vera gel but there was no improvement. He smoked one pack a day since 40 years ago. Intraoral showed an irregular indurated endophytic mass with an ulcer on the right lateral of the tongue. An incisional biopsy was performed and the histopathological diagnosis was poorly differentiated OSCC. The patient was referred to the oncology department and advised for chemotherapy 6 cycles per 3 weeks. He also used herbal medicines containing Panax ginseng radix, Panax pseudoginseng radix, Ophiopogonis japonicus, Ganoderma lucidum, Ligusticum wallichii rhizoma, and Atractylodes macrocephala rhizoma. The content of these herbal medicines have anticancer and immunomodulatory effects. After the second cycle of chemotherapy, there was a significant improvement. OSCC management with chemotherapy combined with anticancer herbal medicines can increase the success of therapy and reduce the side effects of chemotherapy.

Effect of Citrullus colocynth fruit extract on pro and anti apoptotic molecules in human melanoma cell line (A375).

Melanoma remains a major cause of death worldwide. Chemotherapy might be used to treat advanced Melanoma which is associated with some side effects. Hence, providing proper treatment or proper employment of lower toxic substances is essential. Citrullus colocynthis, a medicinal plant, seems a potential anticancer herbal medicine against Melanoma via various efficient compounds. Therefore, it is of interest to analyze the effect of Citrullus colocynthis fruit extract on pro and anti apoptotic molecules in the human melanoma cell line (A375). Cell viability test was done using MTT assay. mRNA expression of Bax and Bcl2 was done by real-time PCR. The data was analyzed statistically by one way analysis of variance and Duncan multiple range test with graph prism version 5. p<0.05 was considered significant. Citrullus colocynthis extract caused a marked increase in cell death in a dose-dependent manner. At the end of 48 hours, maximum inhibition (50%) was at 400 and 500µg/ml. The study demonstrated that the effect of Citrullus colocynthis extract has elevated the Bax mRNA expression on human melanoma cell lines. Data shows that Citrullus colocynthis fruit extract has an anticancer effect on the human melanoma cell line (A375) by increasing the expression of Bax and Bcl2 and this could be because of the phytochemicals present in it.

Ethnomedicinal Survey of Indigenous Flora for the Management of Cancer in South-Western, Nigeria

Background: Nigeria as a multi-ethnic and multi-regional nation is blessed with a great diversity of traditional approaches to common diseases of mankind. Thus, ethnomedicinal surveys are key in reconciling long existing therapeutic approaches as bedrocks for drug discovery to ailing diseases of man today. Objective of Study: An ethnomedicinal survey was carried out in Ondo, Ekiti, Ogun, Osun and Oyo States of South-western region of Nigeria where mostly the Yoruba people live; to identify and document medicinal plants used in the management of cancer. Methods: Herbalists, traditional healers, herb sellers, old men and women who have information regarding the use of herbs either by inheritance or by trainings were questioned with the help of a well-structured questionnaire. A sample size of 200 respondents was randomly chosen with the help of their (community) leaders. Data on the local names, methods of preparation and the plant parts used were collected and analyzed to determine their frequency of citation, species use value and informant consensus factor (ICF). Results: A total of 126 plant species (belonging to 112 genera and119 families) used in the treatment of 20 types of cancer were identified. Members of the family Fabaceae (10 species) were the most widely used. It was observed that Bryophyllum pinnatum and Tetrapleura tetraptera were the most frequently cited species for the treatment of cancer. Analysis revealed 14 plant parts used in the treatment of cancer by the practitioners. Result also showed that bone and kidney cancers had highest degree of consensus from informants (0.5each). Conclusion: Bioassay guided fractionation is required on the most frequently cited plant species such that adequate information based on ethnomedicinal practice for the treatment of cancer in South-Western Nigeria can be kept for successive generation. New anti-cancer herbal medicines may also be discovered to alleviate the suffering of mankind.

Cytotoxic activities of extract and fraction of agarwood leaves (aquaria malaccensis Lam) as herbal medicine on MCF-7 cancer cells

MCF-7 cell is a cell line widely used in studied breast cancer. Breast cancer is one type of cancer that has a high prevalence in the world, including Indonesia. Agarwood leaves (Aquilaria malaccensis Lam) is one of the plants known to possess antioxidant activity. This study aims to determine the cytotoxic activity of extracts and fractions of Agarwood leaves (Aquilaria malaccensis Lam) against MCF-7 cancer cells. Agarwood leaves (Aquilaria malaccensis Lam) were extracted using the maceration method with 96% ethanol and followed by fractionation using n-hexane, ethyl acetate, and methanol: water as a solvent. Phytochemical screening test results Agarwood leaves extract (Aquilaria malaccensis Lam) contains alkaloid compounds flavonoids, quinones, tannins, saponins, steroids and triterpenoids. The results of thin-layer chromatography monitoring showed that the extract and fraction of Agarwood leave hexane had active flavonoid compounds. The extracts and its fractions were tested for cytotoxic activity against MCF-7 cell line using MTT Assay [3- (4,5 dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide]. The results showed IC50 values for extract, methanol:water fraction, ethyl acetate fraction, and n-hexane fraction were 728.412 µg/mL, 505.026 µg/mL, 1881.482 µg/mL, and 144.458 µg/ mL, respectively.The n-hexane fraction of Agarwood leaves (Aquilaria malaccensis Lam) has the best cytotoxic activity against MCF-7 cancer cells and has the potential to be developed as an anti-cancer herbal medicine.

MTT-BASED CYTOTOXIC EVALUATION OF AMBONESE BANANA STEM SAP (Musa paradisiaca var. sapientum (L.) Kuntze) ON FIBROBLAST CELLS

Traditional herbal remedies are naturally occurring, plant-derived substances with limited to no chemical processing and have been used in local or national healing traditions to treat illness. In global health debates, traditional herbal medicines are gaining considerable attention. Many hope new research into herbal medicine will play a vital role in global health. Countries like China, India, Nigeria, USA, and also the World Health Organization (WHO) made large investments in ancient herbal medicines. Currently, the use of the Indonesian plant has dramatically improved the medical and dentistry field. The dental and oral care is often related to wounds, and the Ambonese banana stem has been proven as an effective treatment for these injuries. This study aimed to evaluate the effectiveness and safety of the Ambonese banana stem sap through cytotoxicity tests on the fibroblast cell culture of Baby Hamster Kidney-21 (BHK-21). This study was carried out on three BHK-21 fibroblast cell culture, namely, the media and cell control, and the Ambonese banana stem sap with a concentration of 10%, 20%, 30%, 40%, 50%, 60 %, 70%, 80%, 90%, and 100% incubated for 24 hours at 37ºC and 5% CO2. Then, MTT was evenly dispersed on the media to obtain accurate optical density value. All quantitative data were statistically analyzed using one-way ANOVA and Tukey's HSD Test. The result showed significant differences in optical density values between groups with p = 0.000 (p 0.05). There was no significant difference between the cell control and the Ambonese banana stem sap group with concentrations of 90%, 80%, 70%, 60%, 50%, 30%, 20%, and 10%. It was also observed that the Ambonese banana stem sap is nontoxic to fibroblast cells since its viability value was more than 60%.

None-Loss Target Release of Biomimetic CaCO3 Nanocomposites for Screening Bioactive Components and Target Proteins

Many traditional Chinese medicines have been clinically proven to have good anticancer effects in recent years. Tripterygium wilfordii, as a classic anticancer herbal medicine, has good effects, bu...

Low-Dose Triptolide Promotes MDM2 Inhibitor Nutlin3a to Induce Acute Myeloid Leukemia Cell Death Via p53-Dependent and -Independent Mechanisms

The tumor suppressor p53 is central to hematopoietic stem cell function. Loss of p53 function is associated with poor prognosis of acute myeloid leukemia (AML). P53 negative regulator MDM2 is frequently overexpressed in AML, thus it becomes an attractive therapeutic target for the treatment of AML with wild-type p53 (Wt-p53). Targeting MDM2 to restore p53 activity has been assessed in AML, but clinical outcomes are less promising. Notably, p53-mutated AML still lacks effective treatment approaches. Therefore, our study strived to explore potent therapeutics to target AML with p53 wild-type and mutations. Triptolide (TPL), an ancient herbal medicine, has been used to treat immune disorders for centuries. Accumulating evidence including ours have proven that low dose of TPL potentiates the efficacy of chemotherapies in a broad range of tumor types. In this study, we evaluated the anti-leukemic efficacy of combining low dose TPL with Nutlin3a, a MDM2 inhibitor, against AML with p53 wild-type and mutations. We found that low-dose TPL synergized with Nutlin3a to induce mitochondrial apoptosis in AML cells with Wt-p53 bothin vitroandin vivo. Underlying mechanism for the synergy showed that Nutlin3a upregulated pro-apoptosis factors (e.g. PUMA, p21), while low-dose TPL suppressed MDM2 transcription and reduced anti-apoptosis proteins (e.g. XIAP, Mcl-1) in Wt-p53 AML cells. Interestingly, we also observed that TPL but not Nutlin3a induced cell death in p53 shRNA knockdown, p53 deficiency cell lines (e.g. THP-1, HL-60) and primary samples. Thus, the p53-independent role of TPL was consequently focused. Our RNA-Seq analysis identified 621 differential expression genes (DEGs) after TPL treatment. These genes, including MYC, ATF4 and 4EBP1, were enriched in intrinsic apoptosis pathway responding to ER stress and thus we concluded that Triptolide played its p53 independent role via MYC-ATF4 axis. Collectively, these findings suggest that coadministration of Nutlin3a with low dose TPL would benefit for AML patients. Disclosures Carter: AstraZeneca:Research Funding;Syndax:Research Funding;Ascentage:Research Funding;Amgen:Research Funding. OffLabel Disclosure: Triptolide (TPL), an ancient herbal medicine, is applying to treat immune disorders.

Cytotoxicity evaluation of curcumin-loaded affibody-decorated liposomes against breast cancerous cell lines

Curcumin is known as an effective anticancer herbal medicine but unfortunately, its bioavailability is poor which necessitate efforts for developing more efficient and specific delivery systems. Human epidermal growth factor receptor 2 (HER 2) due to its overexpression in various types of cancers, is demonstrated to be a good candidate as a target for anticancer therapy. In this study, cytotoxicity of curcumin encapsulated in ZHER2:342 Affibody-decorated liposome was investigated against SKBR3 and MCF-7 cancerous cell lines. Curcumin-containing liposomes were prepared from soybeans lecetin and cholesterol by thin-film hydration method. Affibody ZHER2:342 molecules via C-terminal cysteine residue were conjugated covalently to the prepared liposomes. Particle size analysis was performed using atomic force microscopy (AFM) and dynamic light scattering (DLS). Curcumin loading was measured using UV-Vis spectrophotometry and cytotoxic activity of curcumin formulations against cancerous cell lines was investigated by MTT assay. Induction of apoptosis was investigated using flow cytometry through Annexin V staining. Particle analysis showed the formation of spherical liposomes with a mean diameter of about 150 nm. Cytotoxic activity of curcumin was improved by its encapsulation in both liposomes and affibody-decorated liposomes. The Annexin V staining indicated the induction of apoptosis by affibody-decorated liposomes in both MCF-7 and SKBR3 cells. Decoration of curcumin-loaded liposomes with affibody ZHER2:342 may improve curcumin apoptotic function independently of HER2 expression level.

Main nutrients and functional ingredients in shoots of Polygonatum cyrtonema

In order to reveal the main nutrients and functional ingredients in the shoots of Polygonatum cyrtonema, the polysaccharides, proteins, amino acids, and total phenols were determined. The tested samples cultured in Ma'nijiaonong, Hengtang village, Tianmushan town, Lin'an, Zhejiang, which were collected from three provenances(Pan'an and Longquan in Zhejiang and Qingyang in Anhui). The results showed that the polysaccharide content of the shoots varied from 2.34% to 12.73%, roughly one-third of rhizomes. The protein content varied from 107.75 to 192.49 mg·g~(-1), nearly 5.50 times more than rhizomes. Moreover, the average of total amino acid content was 193.13-248.74 mg·g~(-1), approximately 4.16 times of rhizomes. And the essential amino acids account for 35.57%-39.44% of the total amino acids content, which was close to the standard of the ideal protein proposed by FAO/WHO(the essential amino acid/total amino acid is about 40%). In addition, the taste amino acids(TaAA) changed from 160.12 to 208.29 mg·g~(-1), revealing the material basis of &quot;shoots were extremely delicious&quot; in Chinese ancient herbal medicine. Additionally, the total phenols varied from 51.21-58.76 mg·g~(-1), about 2.96 times of rhizomes. The DPPH free radical scavenging rate of tested shoots was over 95%, which obviously superior to rhizomes. Therefore, the shoots of P. cyrtonema is a very high-quality vegetable and functional food with good development potential. Furthermore, the main nutrients and functional substances in P. cyrtonema shoots are closely related to the provenances and harvesting seasons. It is important to improve the quality and yield of the shoots by strengthening the variety of breeding and cultivation techniques.

Historical evolution of Panax notoginseng (Sanqi) processing methods

In the folk medicine, it was believed that the raw Sanqi could remove blood stasis, swelling and pain, while the cooked Sanqi could replenish blood. This article began with the first record of Sanqi in ancient herbal medicine literature, and the nature, flavor and efficacy of Sanqi, then listed the processing methods and usage of Sanqi recorded in ancient herbal medicine literature, and compared with the modern processing methods. The results showed that the processing methods of raw Sanqi included making powder, chewing, vinegar grinding and wine grinding, in which the method of making powder was still in use, and there were particle size requirements on the Sanqi powder listed in most modern preparation specifications. The processing methods of cooked Sanqi included baking, frying and steaming, in which the method of steaming was still in use, but had some changes. The new processing method was frying with edible oil. The results clarify the history of the processing of Sanqi, and provide a reference for the research and application of the processing methods of raw and cooked Sanqi.

In vitro and in vivo anti-leukemia activity of the stem bark of Salacia impressifolia (Miers) A. C. Smith (Celastraceae)

Ethnopharmacological relevanceSalacia impressifolia (Miers) A. C. Smith (family Celastraceae) is a traditional medicinal plant found in the Amazon Rainforest known as “miraruíra”, "cipó-miraruíra" or “panu” and is traditionally used to treat dengue, flu, inflammation, pain, diabetes, male impotency, renal affections, rheumatism and cancer. Aim of the study: The aim of this study was to investigate in vitro and in vivo anti-leukemia activity of the stem bark of S. impressifolia in experimental models. Materials and methods: The in vitro cytotoxic activity of extracts, fractions and quinonemethide triterpenes (22-hydroxytingenone, tingenone and pristimerin) from the stem bark of S. impressifolia in cultured cancer cells was determined. The in vivo antitumor activity of the ethyl acetate extract (EAE) and of its fraction (FEAE.3) from the stem bark of S. impressifolia was assessed in C.B-17 severe combined immunodeficient (SCID) mice engrafted with human promyelocytic leukemia HL-60 cells. Results: The extract EAE, its fraction FEAE.3, and quinonemethide triterpenes exhibited potent cytotoxicity against cancer cell lines, including in vitro anti-leukemia activity against HL-60 and K-562 cells. Moreover, extract EAE and its fraction FEAE.3 inhibited the in vivo development of HL-60 cells engrafted in C.B-17 SCID mice. Tumor mass inhibition rates were measured as 40.4% and 81.5% for the extract EAE (20 mg/kg) and for its fraction FEAE.3 (20 mg/kg), respectively. Conclusions: Ethyl acetate extract and its fraction from the stem bark of S. impressifolia exhibit in vitro and in vivo anti-leukemia activity that can be attributed to their quinonemethide triterpenes. These data confirm the ethnopharmacological use of this species and may contribute to the development of a novel anticancer herbal medicine.