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: 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.