Introduction: Stinging nettle (Urtica dioica L.) belongs to the family of Urticaceae. Three species of Urticaceae (Urtica cannabina, Urtica angustifolia, Urtica dioica L) was grown in Mongolia. U. dioica has recently been shown to have antibacterial, antioxidant, analgesic, anti-inflammatory, antiviral, anti-colitis, anticancer and antiAlzheimer activities. Flavonoids, tannins, scopoletin, sterols, fatty acids, polysaccharides, isolectins and sterols are phytochemicals which are reported from this plant. But effect of hair growth is unclear yet. Goal: We investigated the effect of Urtica dioica L extracts on hair growth by using in-vitro and ex-vivo study methods. Materials and Methods: Human single hair follicle and dermal papilla cells obtained from scalp skin samples of healthy volunteers. We evaluated the effect of Urtica Dioica L on hDPCs and on ex-vivo hair follicle organ culture. Hair follicle matrix cell’s proliferation marker Ki-67 identified by immunoflurescence staining. Result: Urtica Dioica L ethanol extracts promoted elongation of the hair shaft and reduced catagen transition of human hair follicles in organ culture model. E.extract of Urtica Dioica L increased Ki-67 positive matrix keratinocytes. Conclusion Urtica Dioica L ethanol extract enhanced human hair growth in ex-vivo organ culture model. Needed future study to investigate the related mechanism of hair growth.

Background: In 2021, 5981 of cancer new cases was registered in Mongolian population. Among those cases, liver cancer was commonly registered with a prevalence of 32.7%. Studies on anticancer agents with no-adverse effects and good-preventive efficacy against cancer have been attracted more attention from the researchers in the field of pharmaceutical sciences. Scutellaria baicalensis Georgi, Saussurrea amara.L, Chiazospermum erectum Berh, and Carthamus tinctorius.L are well recognized as effective agent against liver diseases. Using these raw materials, researchers have been invented a traditional prescription and named as Hepaclin-4. In this study, we aimed to investigate the qualitative study of raw materials and some biologically active sub- stances in the compounds. Purpose: To study the qualitative study of raw materials for Hepaclin-4 prescription Materials and methods: Some qualitative properties of raw materials for Hepaclin-4 prescription, including appearance, minerals, some organic compounds, total ash, water-soluble substances and fungi, were investigated according to Mongolian pharmacopeia and total flavonoid was detected by thin layer chromatography. Results: No changes were observed on the appearance of raw materials, and minerals and organic compounds weren’t detected in the prescription. No contamination with fungi and insects were identified. The moist in the raw materials were 5.9 to 8.1%, total ash was 4.7 to 13.3% and the water-soluble substances were detected 33.8 to 42.9%. Number of aerobic bacteria, fungi and E.coli, Salmonella species were detected in normal range, indicating that the prescription was matched with the requirement of pharmacopeia. According to the thin layer chromatography study of the raw materials, a yellow spot on the chromatogram were identified and same as quercetin (Rf=0.9-0.98) and rutin ((Rf=0.18-0.23)) as standard compounds, which indicated that the spot which indicated that the spot was flavonoids in the prescription. Conclusions These results showed that the appearance, moist, minerals, organic compound, water-soluble substances, ash and biologically active substances of the raw materials for Hepaclin-4 prescription was corresponded with the requirements of pharmacopeia, and flavonoid was detected in raw materials of Hepaclin-4.

Background: Nitric oxide (NO) is a biological messenger molecule that plays a significant role in the pathogenesis of inflammation. It has anti-inflammatory effects under physiological conditions but can act as a pro-inflammatory mediator when produced excessively under abnormal conditions. NO is involved in the pathogenesis of inflammatory diseases affecting the joints, intestines, and lungs. Therefore, compounds that inhibit NO production are considered important for the treatment of inflammatory diseases and are used clinically. The RAW 264.7 mouse macrophage-like cell line is a widely used model for inflammation studies. Lipopolysaccharide (LPS), a component of the outer membrane of Gram-negative bacteria, is used to activate RAW 264.7 cells and create an inflammation model. Glycyrrhiza uralensis, also known as licorice, is a perennial herbaceous plant in the Fabaceae family. It has been widely used in traditional medicine due to its anti-inflammatory, antiviral, and hepatoprotective properties. Recent studies have shown that licorice contains bioactive compounds such as glycyrrhizin, liquiritigenin, and isoliquiritigenin, which play an important role in inhibiting the synthesis of pro-inflammatory cytokines in macrophages induced by LPS. Inula helenium L., also known as elecampane, is a perennial herbaceous plant used as an expectorant, anti-infective, anti-inflammatory, and anti-helminthic agent in various respiratory diseases. Licorice and Inula helenium are included in Mongolian traditional medicine prescriptions, but their anti-inflammatory effects have not been fully determined, which forms the basis for this research. Aim: To study the effect of Glycyrrhiza uralensis and Inula helenium extracts on the production of NO, the end product of inflammation, in RAW 264.7 macrophage cell lines stimulated with lipopolysaccharide. Materials and Methods: The non-toxic dose of the plant extracts was determined in RAW 264.7 mouse macrophage-like cell line cultures using the MTT assay. Nitric oxide production in RAW 264.7 cell line cultures stimulated with lipopolysaccharide was assessed using the Griess method. Statistical analysis of the results was performed using SPSS 25.0 software, with the p-value calculated by one-way ANOVA, and the differences between groups were evaluated. Results: In RAW 264.7 cell cultures, Glycyrrhiza uralensis and Inula helenium extracts were non-toxic and promoted cell growth at doses ranging from 1 to 25 μg/ml, while a dose of 50 μg/ml was toxic and inhibited cell growth (p<0.01). When the combined plant extracts were applied to cells at doses ranging from 1 to 100 μg/ml, they were also non-toxic and enhanced cell growth, while a dose of 500 μg/ml was toxic and inhibited growth (p<0.001). In terms of nitric oxide production, Glycyrrhiza uralensis extract increased NO production in a dose- and time-dependent manner compared to the control or PBS-treated group. However, Inula helenium extract did not show a dose- or time-dependent effect on NO production. In the lipopolysaccharide-induced inflammation model, licorice extract inhibited NO production at a dose of 30 μg/ml after 12 hours, and further reduced NO production in a dose- and time-dependent manner after 48 hours. Conversely, no significant changes were observed in the Inula helenium extract group at a dose of 25 μg/ml after 48 hours, but a reduction in LPS-induced NO production was observed at a dose of 25 μg/ml after 48 hours. Conclusion Glycyrrhiza uralensis extract alone increased NO production in a dose- and time-dependent manner. It also reduced LPS-induced NO production in a dose- and time-dependent manner. In contrast, Inula helenium extract inhibited LPS-induced NO production at a dose of 25 μg/ml after 48 hours.

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.

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: Toll like receptors (TLRs) are a class of proteins that key role in the innate immune system. TLR7 is expressed on monocytes, macrophages and dendritic cells, T cell, B cell and eosinophiles. TLR7, originally identified as recognizing imidaquinoline, loxibrine, broprimine and ssRNA, ssRNA viruses such as vesicular stomatitis virus, influenza A virus and human immunodefiency virus. It is known that virus ssRNA affects signaling molecule of IFN-y. Objective: To determine gene and protein activation of IFN-y signal transduction by TLR7 ligand in the endothelial cells. MATERIAL: In study we used mouse aortic linear endothelial cell which is cultured (END-D) in 5% heat- inactivated fetal calf serum (FCS), medium (DMEM) containing antibiotic mix(penicillin G, streptomycin, amphotericin B) at 37°C (5% CO2). Endothelial cells treated with synthetic IFN-γ and imiquimodligands, then the NO (nitric oxide) concentration in the supernatant is determined by Griess reagent. Endothelial cells are cultured in 6 well cell culture plate and in each well 2*104cells are expected to be grown for 24 hours of culture. Then, the cells are treated with synthetic IFN-γ and имиквимод ligand for 6 hours and the NO signaling gene activation iNOS mRNA expression which is induced by IFN-γ is determined by RT-qPCR. Endothelial cells are cultured in 12 well cell culture plate and in each well 2*104 cells are expected to be grown for 18 hours of culture. Then, the cells are treated with synthetic IFN-γ and imiquimodligands for 24 hours and the NO signaling protein iNOS expression which is induced by IFN-γ is determined by western blotting. The experiment was conducted as representation mean of at least three test results. The difference between statistical probabilities is determined by the “Students” t test. The p<0.01 value is assumed to be statistically different. RESULTS: TLR7 ligand imiquimodaugmented interferon gamma induced nitric oxide production TLR7 ligand imiquimodincreased interferon gamma induced iNOS mRNA gene expression. TLR7 ilgand imiquimodup-regulated interferon gamma induced iNOS protein expression. CONCLUSIONS: TLR7 ligand imiquimod augments IFN-γ signaling in the endothelial cells. This synergistic effect has revealed in the levels of gene and protein expression.

Introduction: Toll like receptors (TLRs) are a class of proteins that key role in the innate immune system. The SOCS1 and SHP2 proteins are negative-feed loop inhibitors of signaling of JAK/STAT and TLRs pathways. Purpose: To determine negative regulator protein activation which is activated through TLR7 ligand/IFN-γ signal transduction in endothelial cells. Methods: We used mouse aortic linear endothelial cell (END-D); protein expressio was detected by western blotting Results: We analyzed a time dependent stimulation effects of negative regulator proteins stimulated by TLR7 ligand/IFN-γ in endothelial cell cultures. Imiquimod of 10 μg/ml treatment of 1 hr was followed by 100 ng/ml IFN-γ stimulation for 1-8hr to analysis of negative regulator SOCS1 and SHP2 protein expression.
In untreated cells, there was low activations of negative regulator SOCS1 and SHP2 proteins. IFN-γ stimulation alone had increased SOCS1 and SHP2 protein expressions, also imiquimod treatment highly elevated SOCS1 and SHP2 expressions. However imiquimod and IFN-γ doubled treatment have decreased activation of negative regulator SOCS1 and SHP2 proteins. These findings suggest SOCS1 and SHP2 proteins are inhibitors in the TLR7 ligand/IFN-γ signaling. Conclusion Negative regulators, SOCS1 and SHP2 strongly suppressed activations of TLR7 ligand/IFN-γ signaling

Introduction: The aim of this research project is to elucidate the crosstalk of innate and adaptive immune reactions against the DNA containing bacteria. : This study held in the Core laboratory, Science Technology Center, Mongolian National University of Medical Sciences (MNUMS). Murine aortal endothelial cells, END-D cultured and the cell viability checked by MTT assay. In addition, the NO production, protein and gene expression studied by Griess Reagent assay, R.T-PCR and immunoblotting, respectively. Results: 0.1µM, 1µM and 10µM of TLR9 ligand exhibited no cytotoxic action against the cells by MTT assay. IFN-ү alone induced NO production in END-D cells. In the other hand, TLR9 ligand at 0.1µM, 1µM and 10µM up-regulated IFN-ү induced NO production in dose dependent manner. RTPCR results exhibit that TLR9 ligand up regulates iNOS mRNA. Immunoblotting analysis showed the enhanced iNOS protein expression and phosphorylation of STAT1 in cells pre-treated with TLR9 ligand. Discussion: We have demonstrated CpG DNA, TLR9 ligand, up-regulates IFN-ү induced NO via enhanced IFN-ү signaling. The result of Western Blotting and RT-PCR support the up-regulation of NO. CpG DNA can be used as agent against virus and bacteria. Further research need to be conducted. Conclusion TLR9 ligand, CpG DNA up-regulates IFN-ү induced NO production in time and dose dependent manner. TLR9 ligand augments the expression of iNOS mRNA and STAT1 phosphorylation in response to IFN-ү.

Introduction: When human body encounters external pathogens primary/innate immunity cells are activated by recognizing them and secondary/adaptive immunity is activated consecutively. 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 regulator proteins on the IFN-γ/TLR9 signaling pathway. Methods: In this study, murine endothelial cell (END-D) culture was used. END-D cells pre-treated with TLR9 ligand CpG DNA and then stimulated with IFN-γ. The negative (SHP-2, SOCS1, PIAS1) and positive (p38) regulator protein expression was detected by Western blotting. Results and Conclusion Treatment by TLR9 ligand CpG DNA and IFN-γ increased positive regulator p38 phosphorylation in 0.5 hour. CpG DNA inhibited IFN-γ negative regulator PIAS1 protein expression in 6 hour and SOCS1 and SHP-2 expression could not affect in 4 hour.