Background: Currently, colorectal cancer (CRC) ranks as the third most common cancer and the second leading cause of cancer-related mortality worldwide. CRC frequently metastasizes to the liver (50%), lungs (10–15%), peritoneum (4%), bones (10.7%–23.7%), brain (0.3%–6%), and spinal cord. Approximately 35% of CRC cases are diagnosed before distant metastasis, 36% upon lymph node involvement, and 23% after distant organ metastasis. Although several studies have established primary tumor models in mice in our country, there are limited studies on experimental lung metastasis models, prompting the need for this research. Aim: To establish and evaluate a lung metastasis model of colorectal cancer in C57BL/6J mice using the MC38 cell line. Materials and Methods: This study was conducted at the Institute of Biomedical Sciences, Mongolian National University of Medical Sciences. Approval was obtained from the Ethics Review Board of the Mongolian National University of Medical Sciences (2023/3-09) and all laboratory safety regulations and protocols were strictly followed. Male C57BL/6J mice bred at the Experimental Animal Center of Mongolian National University of Medical Sciences were used. MC38 murine colorectal carcinoma cells were cultured and injected intravenously (via the tail vein) at a concentration of 0.25×10⁶ cells per mouse (n=12) to induce lung metastasis. Histological analysis was subsequently performed. Results: Histological examination revealed significant alterations in lung tissue architecture, characterized by areas of dense infiltration by pleomorphic, hyperchromatic cells, disrupting the normal alveolar structure. No histological abnormalities were observed in other organs. Conclusion Intravenous injection of MC38 colorectal adenocarcinoma cells into the tail vein of C57BL/6J mice successfully induced lung metastases, characterized by hyperchromatic, pleomorphic cell infiltrates forming glandular structures within the lung parenchyma.

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: Medicinal herbs have been used in traditional medicine to treat systemic inflammatory disease for many years. For instance, Rhodiola rosea L extracts were used to enhance behavioural stresses for improving fatigue and depression. Gallic acid, found in Rhodiola rosea L and Rhodiola quidrifida is, a natural polyphenol, exhibits multiple therapeutic activities, including anti-inflammatory, anti-cancer, antioxidant, and anti-angiogenic effects. Saposhnikovia divaricate (its ethanol extract mainly) notified to support function of musculoskeletal tissue and to enhance tissue regeneration by its anti-inflammatory effect. The Salsola laricifolia L has been studied for its strong antioxidant activity, improve immune function, boost energy, and exhibit anti-inflammatory effects. This study allowed us to screen anti-inflammatory effects of medicinal plants and future therapeutic possibility. Aim: To study the content of phenolic compounds (gallic acid) in the composition of complex products of plants widely used in Mongolian traditional medicine, which have been found to have biological activity of the extracts Materials and Methods: The herbal extract was extracted from Rhodiola rosea L, Saposhnikovia divaricata (Tuscz) Schischk, Rhodiola quidrifida Pall.Fisch, Salsola laricifolia Turcz.ex Litv in laboratory of the Drug Research Institute of Monos Group, Mongolia. We used solvents for HPLC grade was used to identify the bioactive components that Gallic acid in the compound plant extract. : Results: The bioactive compounds in each of the Rhodiola rosea L, Rhodiola quadrifida Pall.Fisch and four medicinal plant extracts were identified using HPLC, confirming the presence of Gallic acid. Conclusion Our research results showed that the quantitative content of Gallic acid in the composite plant extract was high, at 1.02%.

Introduction: Scutellaria baicalensis Georgi, which is used in traditional medicine, has the ability to remove blood-drying heat. Chiazospermum erectum Bernh. has the ability to relieve typhoid fever and poison fever. Carthamus tinctorius L. has antiseptic, analgesic and anti-toxic properties. Saussurea amara L. has bactericidal, anti-infective, and anti-inflammatory properties. Researchers found that the Hepaclin-4 recipe has antioxidant, membrane-strengthening, liver-protective, necrosis-preventing, detoxification, and peroxidation product accumulation-reducing properties. Therefore, extracting the granular medicine form from the concentrated extract containing the Hepaclin-4 formulation is the basis of our research work. Goal: To obtain the granular medicine form from the concentrated extract containing ingredients of the Hepaclin-4 recipe. Materials and Methods: The research was carried out with the support of the Institute of Pharmaceutical Research and the University of Pharmaceutical Sciences. The raw materials for the Hepaclin-4 formula were extracted by remaceration with water, 40% ethanol, and 70% ethanol (1:10 ratio). Six types of granules were extracted from the concentrated extract using several excipients by the wet granulation method, and the pouring weight and flowability were determined. Results: The quality index of the concentrated extract of the Hepaclin-4 recipe complies with the standards outlined in the 11th Pharmacopoeia of the National Pharmacopoeia of Mongolia. In qualitative analysis of total flavonoid, spots were detected at the same level as standard quercetin (Rf=0.88) and rutin (Rf=0.4), indicating the presence of flavonoids. According to the results of the above research, lactose was found to be the suitable filler for extracting granules, and starch at 8% was identified as the appropriate binding agent from the concentrated extract of the Hepaclin-4 formula. Conclusion It was found suitable to select 8% lactose as a filler and starch as a binding agent from the concentrated extract of the Hepaclin-4 formula and obtain a granule drug form using the wet granulation method.

Introduction: According to the World Health Organization (WHO) in 2020, brain and central nervous system (CNS) cancers account for 2% of all newly diagnosed cancers in the world and 1.5% in Mongolia. Approximately 85-90% of all brain and other CNS tumors were diagnosed primary brain tumor. In 2019, the average 5 year survival probability was 50% for other cancers and 11% for the primary brain tumors. There were 28 patients with primary brain tumor and 33 relatively healthy individuals in our study. Goal: To study the diagnostic value of serum aquaporin-4 and glial fibrous acidic protein in the diagnosis of primary brain cancer Material and Methods: The Department of Neurosurgery at Third central hospital included 28 patients with primary brain cancer and 33 relatively healthy people. The study was conducted under the permission of the Medical Ethics Review Committee of the Ministry of Health on June 19, 2019 №119. Serum aquaporin-4 and glial fibrous acidic protein content was determined by the ELISA kits method using the human aquaporin-4 and glial fibrous acid protein test kit of the Chinese company “Sanlong”. The level is assumed to be true if the p value is less than 0.05. Results Mean age of the all participants was 42.9±16.5, 64% female and 36% male. Serum aquaporin-4 protein levels were 175.71±13.3 pg/ml and serum glial fibrilliary acidic protein levels were 2.682±0.218 ng/ml in patient with primary brain tumor. Serum aquaporin-4 protein and glial fibrilliary acidic protein levels were statistically significant high (p<0.001) in patient with primary brain tumor. Serum aquaporin-4 protein and glial fibrilliary acidic protein level differences were statistically significant (p<0.05) in benign and malignant tumor. There was no statistically significant correlation between serum aquaporin-4 and glial fibrillary acidic protein level and primary brain tumor grade.

Introduction : In recent years, there has been a significant increase of cerebrovascular disease in Mongolia, which is the second leading cause of mortality. There are dozens of Mongolian traditional medicine which is good efficiency for cerebral ischemia that contains musk. Aim: Therefore, we aimed to investigate the effect of musk under the cerebral ischemia/reperfusion in rats. Materials and Methods: Cerebral middle cerebral artery occlusion was established in male rat (90-minute occlusion followed by 24-hour reperfusion). Rats were divided into following groups: control group, ischemia group (cerebral ischemia and reperfusion), nimodipine administrated group (cerebral ischemia and reperfusion + treated with nimodipine), musk administrated group (cerebral ischemia and reperfusion + musk 50 mg/kg and 100 mg/kg). The brain tissue levels of IL-1β, TNF-α, IL-6, IL-10 cytokines were measured using enzyme linked immunosorbent assay (ELISA) every 1, 3, 7th days. Results: Levels of cytokines (IL-1β, TNF-α and IL-6) were significantly lower in musk treated group compared to brain ischemia group (p<0.05). In contrast, treatment with musk was significantly improved neurological function with stimulation of M2 phenotype microglia cells and increased the anti-inflammatory cytokine level of IL-10 in the ischemic hemisphere of brain in rats Conclusion The mechanisms of musk are associated with increasing the brain tissue levels of IL-10, and reducing the levels of proinflammatory cytokines such as IL-1β, TNF-α, IL-6 subsequently stimulating neurogenesis and reduced ischemic zone. Musk may have neuroprotective effects against cerebral ischemia with stimulating M2 phenotype microglia cells in the brain. Regarding the ELISA, the effects of musk may be due to anti-inflammatory properties through inhibition of some of proinflammatory cytokines and stimulation of anti- inflammatory cytokines.

Background: Establishment of quantitative reference intervals of white blood cells and its subpopulations using a high accuracy analytic system is essential for clinical medicine, public health, and anthropology. We are unable to identify peer-reviewed literature sources describing white blood cell counts and their subpopulations using monoclonal antibodies to specific surface antigens in healthy Mongolians. This study aimed to measure the counts of white blood cells and their subpopulations in healthy Mongolians using flowcytometry. Materials and Methods: The absolute number (cell/L) of leukocytes (CD45+), granulocytes, monocytes and lymphocytes were measured by Magnetic Activated Cell Sorting Assay (MACSQuant Analyzer 10) in 287 blood donors (158 males and 129 females) 17-64 years of age (mean age 33.1±12.4). Peripheral blood samples were collected at the time of blood donation at the National Center for Transfusion Medicine. Results The mean values of leukocytes and granulocytes were lower in donors over 30 years of age (ANOVA: F=4.408, p=0.002 and F=5.685, p=0.001) and regression analysis demonstrated indirect correlation between counts of these cells and age of donors (r= - 0.198, p=0.001 and r=-0.221, p=0.001, respectively). Gender-related differences in white blood cell counts were not found.
Mean value of lymphocyte count in donors investigated in spring (May and March, n = 87; 2224.6±775.3) was significantly higher than those in winter (December – February, n=180; 1613.2±454.3, p=0.001) and autumn (October, n=20; 1576.1±438.6, p= 0.001).
Comparing of our findings with the data from available literature shown that healthy Mongolians have lower leukocyte count compared with Koreans, Chinese Han population and lower mean value of lymphocyte count comparing with Korean, Chinese Han population, and Arabian (Saudi Arabia) populations.

Background: The effect of lipopolysaccharide (LPS) on valproic acid (VPA)-induced cell death was examined by using mouse RAW 264.7 macrophage cells. Materials and methods, results: LPS inhibited the activation of caspase 3 and poly (ADP-ribose) polymerase (PARP) and prevented VPA-induced apoptosis. LPS inhibited VPA-induced p53 activation and pifithrin-α as a p53 inhibitor as well as LPS prevented VPA-induced apoptosis. LPS abolished the increase of Bax/Bcl-2 ratio, which is a critical indicator of p53-mediated mitochondrial damage, in response to VPA. The nuclear factor (NF)-κB inhibitors, Bay 11-7082 and parthenolide, abolished the preventive action of LPS on VPA-induced apoptosis. A series of toll-like receptor (TLR) ligands, Pam3CSK4, poly I:C, and CpG DNA as well as LPS prevented VPA-induced apoptosis. Conclusion Taken together, LPS was suggested to prevent VPA-induced apoptosis via activation of anti-apoptotic NF-κB and inhibition of pro-apoptotic p53 activation.

Introduction: Valproic acid (VPA) has been used in the treatment of seizures and bipolar disorders. In the present study, we examined how VPA affected PI3K-Akt pathway in response to LPS by using mouse RAW 264.7 macrophage cells. Material and methods: Mouse RAW 264.7 macrophage-like cells cultured and the cell viability checked by MTT and TUNEL assay. In addition, protein expression and protein interaction were detected by immune blotting and immune precipitation, respectively. TLR4 expression on cell surface studied by FACS analysis. Results: The MTT and TUNEL assays demonstrated no significant difference between VPA at 2 mM treated and untreated control cells. VPA attenuated LPS-induced phosphorylation of phosphatidylinositol 3-kinase (PI3K) and Akt, but not nuclear factor (NF)-κB and mitogen activated protein kinases (MAPKs). There was no significant difference in the TLR4 expression on the cell surface between cells treated with or without VPA. VPA inhibited LPS-induced PI3K/Akt signal transduction in a dose dependent manner. Conclusion VPA at 2mM exhibits nontoxic effect in the RAW 264.7 cells. VPA down regulates LPS-induced phosphorylation of Akt via inhibition of PI3K activation.

Introduction: When human body encounters external pathogens primary/innate immunity cells are activated by recognizing them and secondary/adaptive immunity is activated consecutively. Immune cell surface receptors, called Toll-like receptors (TLRs) recognize and bind pathogens. In our previous study, we revealed that there is a synergistic action between TLR9 and IFN-γ signaling in the endothelial cells. Purpose: To determine the role of negative and positive regulatory proteins on the IFN-γ/TLR9 synergistic signaling pathway Materials and Methods: This study was held in the Core Laboratory, Science Technology Center, Mongolian National University of Medical Sciences (MNUMS). In this study, murine endothelial cell (END-D) culture was used. The negative and positive regulator protein expression was detected by Western blotting. Result: Result of immunoblotting assay indicated that CpG DNA enhanced IFN-γ positive regulator protein p38 phosphorylation in the endothelial cells. Treatment by TLR9 ligand CpG DNA and IFN-γ increased p38 activation in 0.5 hour and 1 hour. CpG DNA inhibited IFN-γ negative regulator SOCS1 protein expression in 4 hr and 8 hr. Therefore, TLR9 ligand CpG DNA increased IFN-γ signal transduction in the endothelial cell line. Conclusion TLR9 ligand CpG DNA has decreased IFN-γ negative regulator protein SOCS1 expression. CpG DNA has increased IFN-γ positive regulator protein p38 phosphorylation.