Background: Skin wounds can occur for many reasons, including UV radiation, mechanical burns, trauma, and chronic inflammation caused by metabolic factors. In Mongolia, as of 2022, household burns were the top 5 causes of injuries, the leading cause of injuries in children aged 0-5, and accounted for 18.4% of all illnesses in children aged 0-5. Skin burns and wounds take a long time to heal, which causes emotional, aesthetic, and economic stress for doctors. Developing in novative, effective, and low-side effects therapeutic products to improve wound healing is of great clinical and economic importance. The lack of research on the effects of bacterial metabolites and aloe vera (Aloe barbadensis Miller) on burn wounds was the basis for our research. Aim: To determine the effects of a mixture of bacterial metabolites and aloe vera on burn wounds in experimental animals. Materials and Methods: Eight-week-old BALB/c mice were divided into three groups: control group, bacterial metabo lite group, and aloe vera group. The mice were anesthetized, and the dorsal hair was shaved over a 2×2 cm area using an electric clipper. A second-degree burn wound model was created by exposing the shaved dorsal skin to hot air at 200°C for 10 seconds. A total of 100 μL of a bacterial metabolite suspension mixed with aloe vera at a 1:1 ratio was applied evenly to the center of the wound. Wound images were captured daily, and wound areas were measured using ImageJ software. Healing was evaluated on days 7, 14, and 21 of treatment and compared with the control group. The initial wound size was defined as 100%, and wound closure percentages were calculated accordingly. Burn skin tissues from both experimen tal and control groups were subjected to immunofluorescence staining for the vascular marker α-SMA (Thermo Fisher Scientific). Statistical analysis: The differences between the control and experimental groups were analyzed using an independent t-test using GraphPad Prism 10 software, and the mean, standard deviation, and statistical correlation were calculated. Results: The effect of the mixture of bacterial metabolites and aloe vera (Aloe barbadensis Miller) on the healing of burn wounds in experimental mice was compared with the control group. The wound size at 7 days was 82.1% in the control group and 83.7% in the experimental group (p<0.197), at 14 days was 62.6% in the control group and 58.1% in the exper imental group (p<0.0001), at 21 days was 50.7% in the control group and 32.7% in the experimental group (p<0.0001). No statistically significant difference in wound healing was observed between the groups at day 7; however, significant differences were observed at days 14 and 21. Immunofluorescence analysis revealed uniform expression of α-SMA-pos itive cells in the experimental group at 7, 14, and 21 days of treatment, whereas α-SMA expression was not detected in the control group. Conclusion The combination of bacterial metabolites and aloe vera promotes wound healing in experimental mouse burn wounds.

Background: Plants rich in essential oils belong to more than 50 families, including Asteraceae, Rutaceae, and Lamiaceae. These plants are widely used in food, traditional and modern medicine, perfumery, cosmetics, and pharmaceutical industries. Approximately 3,000 types of essential oils have been identified, of which more than 300 are utilized for industrial and commercial purposes. Essential oils exhibit antimicrobial, anti-inflammatory, anticancer, cytotoxic, and antioxidant activities and are therefore widely applied in medical research. In Mongolia, the extreme climate, distinct ecological regions, and the prevalence of pastoral livestock husbandry provide a scientific basis for investigating essential oil–containing plants, their chemical composition, and biological activities. Among them, Juniperus Sabina L. is of particular theoretical and practical interest. Aim: To determine the non-toxic dose of essential oil extracted from Juniper Sabina L. on neural cells. Materials and Methods: This study was conducted at the Institute of Bio-Medicine, MNUMS. Ethical approval was obtained from the Research Ethics Committee of MNUMS (Meeting date: November 21, 2025; Protocol No. 2025-26/0301). BV2 microglial and N2a neuronal cell lines were cultured under standard conditions at 37°C in a 5% CO₂ incubator. Cells were maintained in appropriate culture media, and their growth and morphology were monitored microscopically before experimentation. Cell viability was assessed using the MTT assay. BV2 microglial cells were seeded into 96-well plates at a density of 2×10⁴ cells/well and incubated for 24 hours. Essential oil was added at concentrations ranging from 50–500 µg/mL and incubated for an additional 24 hours. After treatment, MTT reagent was added and incubated for 3 hours. The resulting formazan crystals were dissolved in dimethyl sulfoxide (DMSO), and absorbance was measured at 630 nm. Cell viability was calculated based on absorbance values to determine the non-toxic concentration of the essential oil. Result: The cytotoxic effects of the essential oil were evaluated in BV2 microglial and N2a neuronal cells using the MTT assay. In N2a neuronal cells, concentrations of 50, 100, 200, and 300 µg/mL did not significantly affect cell viability, whereas 400 and 500 µg/mL reduced cell viability, indicating cytotoxic effects. Conclusion According to MTT assay results, the essential oil of Juniperus sabina showed no cytotoxic effect on N2a neuronal cells at concentrations of 50, 100, 200, and 300 µg/mL, while cytotoxic effects were observed at 400 and 500 µg/mL. In BV2 microglial cells, concentrations of 50, 100, and 200 µg/mL were non-toxic, whereas concentrations above 300 µg/mL exhibited cytotoxic effects.

Background: Globally, research on identifying bioactive compounds from medicinal plants and characterizing their in vitro and in vivo pharmacological activities is progressing rapidly. Plants native to Mongolia have adapted to unique, extreme continental climatic conditions, accumulating distinct phytochemicals, and have historically been utilized in traditional medicine and dietary practices. Establishing the non-cytotoxic safety threshold of these wild and cultivated plant extracts in RBL-2H3 mast cells a universally accepted model for type I hypersensitivity and inflammation is a vital prerequisite. Determining these baseline non-toxic dose ranges provides the essential scientific foundation for subsequent cellular allergy models, animal studies, and the elucidation of molecular mechanisms underlying their therapeutic efficacy. Aim: To determine the cytotoxicity profile and establish the non-cytotoxic dose ranges of extracts from four selected Mongolian plant species (wild and cultivated) in RBL-2H3 cells. Materials and Methods: Plant samples including lingonberry (Vaccinium vitis-idaea), strawberry (Fragaria orientalis), black radish (Raphanus sativus), and blue-berried honeysuckle (Lonicera caerulea L.) were standardly collected, authenticated, and cataloged as herbarium specimens at the Botanic Garden and Research Institute, Mongolian Academy of Sciences. A total of 12 distinct extracts were prepared from plant leaves, stems, and fruits using water and ethanol extraction, followed by concentration via rotary evaporation and freeze-drying. Rat basophilic leukemia (RBL-2H3) cells were maintained in EMEM growth medium and utilized during the exponential growth phase. Cell viability and non-cytotoxic concentration thresholds were evaluated using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. Result: Ethanolic extracts of strawberry leaves, black radish, and honeysuckle leaves (≥6.25 µg/mL), as well as aqueous extracts of honeysuckle fruits (≥6.25 µg/mL) and stems (≥25 µg/mL), exhibited dose-dependent cytotoxicity. Conversely, aqueous extracts of lingonberry leaves (≤200 µg/mL), strawberry leaves (≥100 µg/mL), and black radish (≤200 µg/mL), alongside ethanolic extracts of lingonberry leaves (≥50 µg/mL), demonstrated non-cytotoxic profiles. Furthermore, ethanolic extracts of honeysuckle stems (≤300 µg/mL) and fruits (≥12.5 µg/mL), as well as the aqueous extract of honeysuckle leaves (≤50 µg/mL), significantly enhanced RBL-2H3 cell viability. Conclusion The tested plant extracts influenced RBL-2H3 cell viability in a solvent polarity and plant organ dependent manner. Ethanolic extracts of strawberry leaves, black radish, and honeysuckle leaves, along with aqueous extracts of honeysuckle fruits and stems, demonstrated relatively high cytotoxicity, reducing cell viability by >40% starting at concentrations of 12.5, 25, and 50 µg/mL. In contrast, aqueous extracts of strawberry leaves and black radish, as well as both aqueous and ethanolic extracts of lingonberry leaves, maintained low cytotoxicity at concentrations below 200 µg/mL. Notably, ethanolic extracts of honeysuckle stems and fruits (>12.5 µg/mL) and its aqueous leaf extract (<100 µg/mL) enhanced cell viability, exhibiting significant cell proliferation promoting activity.

Background: Evaluation of genotoxic potential is essential for reducing carcinogenic risk and for assessing the safety of newly developed pharmaceutical products, traditional herbal medicines, and health supplements. Such evaluation plays an important role in protecting human health. Therefore, in Mongolia, it is necessary to investigate the safety of domestically produced pharmaceutical preparations, natural products, and traditional medicines on a scientific basis in accordance with international standards; to develop and implement national policies and standards; to introduce and localize appropriate methodologies; to protect human health; to prevent cancer, hereditary diseases, and longterm adverse effects; and to promote the rational use of medicines. Aim: To investigate the mutagenic properties of an herbal combination preparation in vitro and its genotoxic effects in vivo. Materials and Methods: Extracts were prepared from Dasiphora fruticosa, Cynara scolymus, and Rosa acicularis, and a herbal combination preparation was formulated in specific proportions. Mutagenicity was evaluated according to OECD Guideline 471 (Bacterial Reverse Mutation Test) using the Salmonella typhimurium TA100 strain under both S9 (-) and S9 (+) conditions with the Muta-Chromoplate™ assay. The combination preparation was tested at concentrations of 5, 2.5, 1.25, 0.625, 0.3125, and 0.1562 mg/mL in 96 wells, and reverse-mutated bacterial colonies were assessed by counting the number of wells that changed color from purple to yellow. In addition, using the IPHASE Biosciences kit, the preparation was tested at doses of 5, 2.5, 1.25, 0.625, 0.3125, and 0.1562 mg/plate on Salmonella typhimurium TA100 and TA98 strains under both S9 (-) and S9 (+) conditions. After incubation, the number of bacterial colonies formed was counted and compared with that of the positive control group. Genotoxicity was assessed according to OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test) by evaluating the numbers of polychromatic erythrocytes (PCEs), normochromatic erythrocytes (NCEs), and micronucleated polychromatic erythrocytes in the bone marrow of BALB/c mice. Result: In the study using the Muta-Chromoplate™ assay, bacterial colonies were observed at doses ranging from 5.0 to 0.1562 mg/mL of the herbal combination preparation. However, compared with the positive control group (aminoanthracene), the number of bacterial colonies was significantly lower (p<0.05). In the study using the IPHASE Biosciences kit, 30–154 bacterial colonies were observed in Salmonella typhimurium TA98 and TA100 strains under S9 (-/+) conditions at doses ranging from 5.0 to 0.156 mg/plate, which was approximately threefold lower than that observed in the positive control groups. In the positive control group treated with cyclophosphamide, the number of micronucleated polychromatic erythrocytes was increased. Conclusion The herbal combination preparation showed no mutagenic activity in either of the two mutation detection assays using Salmonella typhimurium TA98 and TA100 strains under S9 (-/+) conditions. In addition, it showed no genotoxic effect in the micronucleus assay.

Background: Stress, insomnia, and anxiety are among the most prevalent mental health disorders in society and are associated with negative consequences such as reduced work capacity, decreased quality of life, and an increased risk of chronic diseases worldwide. During stress, the sympathetic nervous system becomes activated, leading to physiological changes such as increased heart rate, vasoconstriction, and elevated arterial blood pressure. If prolonged, these changes may increase the risk of developing hypertension and cardiovascular diseases. In recent years, pharmacological research on medicinal plants with sedative, hypnotic, and anxiolytic effects has been intensively conducted. Therefore, there is a growing need to develop a combined herbal preparation from Valeriana officinalis L. and Leonurus sibiricus L., which have been used in traditional medicine for sedative, hypnotic, and anxiolytic effects, and to investigate their effects on arterial blood pressure. Aim: To investigate the effect of an herbal complex preparation on arterial blood pressure. Materials and Methods: An herbal complex preparation was formulated from 70% ethanol extracts of Valeriana officinalis L. and Leonurus sibiricus L. in a 30:70 ratio. Its effects on normal blood pressure and Angiotensin II-induced hypertension were evaluated in C57BL/6 mice. The preparation was administered orally at doses of 750 and 1000 mg/kg. Systolic and diastolic blood pressure were measured using a Softron BP-98AWU system and compared between groups. Results: At doses of 750 and 1000 mg/kg, the complex preparation reduced systolic blood pressure (SBP) by 6.4-12.0% and diastolic blood pressure (DBP) by 16.1-28.1% under normal conditions (p<0.05). In the intraperitoneal Angiotensin II-induced hypertension model, SBP decreased by 5.9-19.6% and DBP by 6.3-20.1% (p<0.05). In the osmotic pump-induced Angiotensin II hypertension model, the 750 mg/kg dose reduced SBP by 18.2-33% and DBP by 17.2-28.3% (p<0.001). Conclusion The herbal complex preparation at doses of 750 and 1000 mg/kg demonstrated a significant antihypertensive effect in both normal and Angiotensin II-induced hypertensive mice.

Background: Of the 7,678 plant species growing in Mongolia, more than 1,230 are considered medicinal plants; however, around 900 of these remain unstudied, and only about 200 species are currently used in medical practice. In recent years, the use of natural products in therapy has increased, and research in this field has been rapidly developing. Schizonepeta multifida (L.) Briq., a plant belonging to the Lamiaceae family, is widely distributed in Mongolia, Siberia, and East Asia, and is traditionally used to treat colds, fever, and inflammatory diseases. Phytochemical studies have shown that its essential oil contains terpenoid compounds such as pulegone and limonene, as well as flavonoids. These compounds are known to exhibit antioxidant, anti-inflammatory, and neuroprotective effects; however, comprehensive studies on these activities of Schizonepeta multifida growing in Mongolia are limited. Aim: To determine the antioxidant activity, cytotoxicity, and anti-inflammatory effects of Schizonepeta multifita Briq. essential oil and to evaluate its anti-stress effects in an experimental chronic stress animal model. Materials and Methods: The study was conducted at the Institute of Bio-Medicine, Mongolian National University of Medical Sciences. The aerial parts of Schizonepeta multifida were subjected to hydrodistillation using a Clevenger apparatus to obtain essential oil, and the yield was calculated. Antioxidant activity was measured at 517 nm using a spectrophotometer (Agilent BioTek Epoch 2). Cytotoxicity was assessed in BV2 and N2a cell lines using the MTT assay, after which inflammatory markers (IL-6 and TNF-α) were determined in cell cultures using ELISA. For the induction of chronic stress, C57/BL6 mice were divided into healthy control, negative control, and experimental groups. A total of 10 different chronic stress models were randomly applied twice daily for 28 days. Anti-stress effects were evaluated using the forced swim test. Data analysis was performed using Prism 10 software. Results: The yield of essential oil from Schizonepeta multifida was 0.3±0.03%. The antioxidant activity, assessed by the DPPH radical scavenging method, showed an IC₅₀ value of 262.7±2.95 µg/mL, indicating antioxidant potential. Cytotoxicity assays in BV2 and N2a cell lines showed toxic effects starting from a concentration of 200 µg/mL. The essential oil significantly reduced inflammatory markers IL-6 and TNF-α at concentrations of 50, 100, and 200 µg/mL (p<0.05). In the forced swim test, there was a statistically significant difference (p<0.05) between the disease control group and the experimental group. Conclusion The essential oil of Schizonepeta multifida exhibits antioxidant, anti-inflammatory, and anti-chronic stress effects.

Background: Insects are an inexhaustible source of medicine, and it is essential to study the biologically active substances contained in insect bodies. Although there is information about various insects and beetles used in traditional Mongolian medicine treatment, there are few studies that have compiled and compared these at the source level. Aim: To study and determine from primary sources information about the composition of pheromone secretions contained in the bodies of various insects and worms used in “Entomotherapy (Insect therapy)” in traditional medicine, particularly rare beetle species, and information about what diseases and ailments these insects were used to treat. Materials and Methods: This study used primary source and historical research methods, analysis-synthesis methods, systematic analytical methods, and historical comparative methods. Information was extracted from traditional medicine source books about insect therapy, particularly identifying and compiling primary sources about the use of beetles in folk medical treatment. Results: According to primary sources, traditional Mongolian medicine used quite a variety of “therapeutic insects and worms” of animal origin, including representatives of arthropods such as scorpions, spiders, centipedes, wasps, bees, various types of beetles, and moths. Among these, stag beetles, dung beetles, and black beetles were used in traditional medicine for treating joint and muscle diseases, various women’s diseases, epilepsy and convulsions, headaches, and all 11 types of bile diseases, with specific prescriptions developed for each condition. Conclusion Traditional medicine treatment used various insects from the biological classification of the insect class, particularly many types of beetles from the Coleoptera order. The pheromone secretions contained in their bodies include many different types of chemical substances.

Background: The study of small-molecule compounds with antitumor activity involves several crucial steps. These include determining their selective effects on cancer cells, understanding the type of cell death they induce, identifying the activated signaling pathways, pinpointing the target molecules, and elucidating the mechanisms of action. Among the plant-derived compounds with anticancer properties, flavonoids are notable for their ease of isolation and their abundance in food. Apigetrin, a representative flavonoid, is a secondary metabolite found in plants, and our previous study indicated that its anticancer selectivity index was 13.1. However, the specific mechanism by which apigetrin inhibits leukemia cell growth remains unclear. Aim: To study of the inhibitory action of apigenin on leukemia cell culture Materials and Methods: In this study, we evaluated the apoptosis of cells using flow cytometry and investigated the in volvement of the caspase pathway through the use of pancaspase inhibitors to explore the effects of apigetrin on leukemia cell growth. Results: After incubating leukemia RAW264.7 cells with 30 μM apigetrin for 24 and 48 hours, we did not detect any apoptosis through Annexin V and PI staining by flow cytometry. We compared the number of viable cells using the MTT assay after 24-hour treatment of apigetrin with or without pretreatment of Z-VAD, a pancaspase inhibitor, for 30 minutes. The results indicated that the pancaspase inhibitor did not reduce the inhibitory effect of apigetrin on the growth of RAW264.7 cells. In contrast, the positive control group, treated with doxorubicin—which induces apoptosis—showed not only significant apoptosis but also a reduction of the pancaspase inhibitor on the cell growth inhibition. Therefore, these data suggested that apigetrin likely has a cytostatic effect or inhibits the cell cycle rather than being cytotoxic. Future research should focus on determining which stage of the cell cycle RAW264.7 cells treated with apigetrin are in, as well as studying the signaling pathways involved in the cell cycle. Conclusions Apigetrin inhibits the proliferation of RAW264.7 leukemia cells in a caspase-independent and non-apoptotic manner.

Background: Cancer incidence and mortality are steadily increasing both globally and in Mongolia. As these rates rise, traditional Mongolian medicine has long utilized herbal formulas for the treatment of gastric and esophageal cancers and precancerous conditions. One such formulation—Hot Natured 3 Herbs (HN3H)—comprises three species from the Ranunculaceae family: Atragene sibirica L., Ranunculus repens L., and Pulsatilla bungeana L.. However, scientific validation of its anti-tumor effects is essential. This study aimed to investigate the effect of HN3H in a tumor-induced animal model. Aim: To identify the biologically active compounds of HN3H and evaluate their effect in an experimentally induced tumor model in animals. Materials and Methods: The three herbs comprising HN3H—Atragene sibirica L., Ranunculus repens L., and Pulsatilla bungeana L.—were collected during their flowering stage (May–June) in Khishig-Undur, Bulgan province, and dried according to official procedures. Extraction was carried out by maceration in 96% ethanol at a 1:10 ratio. The concentrated extract was suspended in water (1:1) and successively fractionated with dichloromethane, ethyl acetate, butanol, chloroform, and n-hexane. The study was approved by the Research Ethics Committee of the Mongolian National University of Medical Sciences (Protocol №2020/03-04). A colorectal cancer model was established by subcutaneous injection of MC-38 cells (Kerafast, USA) into C57BL/6 mice. Immunohistochemistry was performed using CK20, CDX2, Ki67, and p53 antibodies at 1:100 and 1:200 dilutions. Results: The ethanol extract of HN3H contained 2.98±0.04% total phenolics and 2.16±0.05% total flavonoids. Body weight and tumor volume were measured daily with three repetitions. All groups showed a time-dependent increase in body weight. Mice in groups 1A and 1B received ethanol extract at 50 and 100 mg/kg doses; groups 2A and 2B received dichloromethane extract at the same doses. The negative control group was administered 0.5 mg/kg PBS orally, while the positive control group received intraperitoneal injections of 5-fluorouracil (5FU) at 10 mg/kg twice a week. Tumor growth increased in a time-dependent manner across groups. Compared to the negative control, tumor volumes in four treatment groups showed statistically significant reduction (p˂0.05), while no significant difference was observed when compared to the positive control (p=0.08). Histological analysis revealed necrosis in all groups, with variation in extent. Conclusion The ethanol extract of HN3H exhibited moderate levels of phenolic compounds and a high concentration of flavonoids. HN3H extract inhibited tumor progression and activated lymphocyte-predominant inflammation in tumor tissues, indicating potential anti-tumor activity (p˂0.05).

Background: According to the World Health Organization (WHO), approximately 70-95% of developing countries rely on traditional medicine, which includes around 365 plant, animal, and mineral-based preparations. Natural products consist of numerous biologically active compounds that exert effects against pathogens, making up about 25% of modern pharmaceuticals derived from plants. Since plants are a combination of various metabolites, they can have therapeutic effects, side effects, and toxicity in the human body. Based on the traditional use of medicinal herbs in Mongolian and Tibetan medicine for their sedative properties, we selected the medicinal herbs Valeriana officinalis L. and Leonurus sibiricus L., The objective is to develop new medicinal preparations that can be utilized in modern medical practice to treat, prevent, or supplement the treatment of depression and anxiety. Consequently, it is necessary to prepare a herbal complex from these selected plants and conduct studies to investigate their subchronic toxicity and sedative activities. Aim: To study subchronic and sedative activity of herbal complex preparations. Materials and Methods: The herbal complex preparation was prepared from the 70% ethanol extract of the roots of Valeriana officinalis L. and the aerial parts of Leonurus sibiricus L., and a compound preparation was made in a 30:70 ratio. Subchronic toxicity study was conducted on Wistar rats weighing 180-250 g according to the OECD-407 guidelines. The sedative activity of herbal complex preparation was studied on C57BL/6 and BALB/c mice using the dark/light transition test according to Takao K., and the hole-board test according to Hiroshi Takeda. Results: In the sub-chronic toxicity study of the herbal complex preparation, biochemical analysis of the serum (including ALT, AST, creatinine, and urea) and histopathological examination of the liver, kidney, and heart showed no statistically significant changes when comparing the experimental groups to the control group. The herbal complex preparation at a dose of 1000 mg/kg increased the time spent in the dark area, decreased the time spent in the light area, and the number of transitions between the two areas of mice in the dark/light transition test, and reduced the number of head-dipping into the holes of mice in the hole-board test. Conclusion The herbal complex preparation exhibited low toxicity at doses of 1000 mg/kg and 1500 mg/kg based on biochemical and histopathological examinations in the subchronic toxicity study. Furthermore, the preparation demonstrated sedative effects at a dose of 1000 mg/kg.