Background: Gliomas are common primary brain tumors of glial origin in adults with a poor prognosis. Tumor-related inflammatory cytokines such as interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and aquaporin-4 (AQP-4) water channel protein have been reported to be expressed in gliomas Aim: This study aimed to determine their serum levels in patients with primary brain tumors compared to healthy controls and to evaluate their potential diagnostic value. Materials and Methods: Serum AQP-4 levels were measured by ELISA in 28 patients with primary brain tumors (PBT) and 26 healthy controls. IL-6 and TNF-α were assessed by flow cytometry in 59 PBT patients and 52 controls. Results: Among the primary brain tumor cases, 45.61% were meningiomas, 26.32% anaplastic astrocytomas, 8.77% glioblastomas, and 12.28% other tumors (including chondroma, ependymoma, schwannoma, craniopharyngioma). Serum IL-6, TNF-α, and AQP-4 levels were significantly higher in the PBT group compared with healthy controls. Conclusion Elevated serum levels of AQP-4, IL-6, and TNF-α in patients with primary brain tumors suggest that these biomarkers molecules could represent valuable non-invasive biomarkers for the early detection of PBT.

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: 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.

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: 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.

BACKGROUND Bovine colostrums is the milk secreted by cows during the first few days after parturition. It contains many essential nutrients and bioactive components, including growth factors, immunoglobulins, lactoperoxidase, lactoferrin and cytokines ets. Lactoferrin has been reported for its multifunctional properties such as antifungal, antibacterial, antiviral antioxidant and anticancer activities. The aims of this study focused on the isolation and purification of lactoferrin from Mongolian bovine colostrums. Lactoferrin purified using HiTrap DEAE an ion exchange chromatography. Lactoferrin purification efficiency was about 60.5%. The single band of purified lactoferrin has been observed in SDS-PAGE electrophoresis. METHODS Bovine colostrum was collected at a cow farm in the Darkhan province of Mongolia. At first the cream was separated by centrifugation (10000 xg 20 min at 4oC). In order to separate the whey, the samples were precipitated with 1mol/l to pH 4.6 and centrifuged at 10000 g 20 min again. The samples of whey were stored at -18oC to the analysis. Lactoferrin was purified by HiTrap DEAE an ion exchange chromatography using 0.005 M phosphate buffer (pH 7.7) and linear gradient NaCl from 0.25M, 0.5M, 1M. During chromatography, protein in the eluents was monitored by ultraviolet absorbation at 280 nm with the instrument. Purity test done by using polyacrylamide gel electrophoresis under denaturated condition (SDS-PAGE) method by Laemmli (1970). For HPLC determination of the lactoferrin by Shimadzu Nexera X2 HPLC system with UV/ VIS detector were used. Detection was carried out at the wavelength 280 nm. Separation was performed on a chromatographic column Protein R C18 ,2.2 x 150 mm, 5 μm particle size. Linear gradient and flow rate 0.2 ml/min were used. Mobile phase a consisted of water / acetonitrile/ trifluoroacetic acid ( 95:5:0.1). The column temperature was set at 40oC and injection volume was 10 μl. Data were collected and evaluated by software Lab Solution. An external standard method for quantification analytes was used. RESULTS Purified lactoferrin in the present study had a good concentration and purification efficiency was about 60.5 %. Protein fraction from 1M NaCl gradient delivers sharp and clean peak to HPLC chromatogram that fits intensity and retention time of standard bovine lactoferrin. Ammount of lactoferrin in bovine colostrums was 0.6 mg/ml and it`s molecular weight 80 kDa as a standard sample. The retention time of lactoferrin fraction which is purified by SDS-PAGE gel electrophoresis. The peak of fraction same compared to the standard lactoferrin 5.8 minutes by HPLC analysis. CONCLUSION Ion exchange chromatography shows reliable and easy isolation of lactoferrin from Mongol bovine colostrum.

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: 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.

BackgroundThere are 137 soums of 17 provinces have plague foci in Mongolia. The 51.7% of them is case, 23.4%- low, 9.5% - high, 0.7% - hyper active. Main host of plague foci is marmot in Mongolia. According last20 year’s surveillance study, about 75.5% of Y.pestis was isolated from marmot, marmot carcassesand their flea. Human plague cases has been caused illegal hunting marmot in Mongolia. Even legaldocument which prohibited marmot hunting was appeared since 2005, people has been hunting marmotfor selling marmot meat, skin and other products. It is depends economy crises and other public issues inMongolia. Also influenced increase risk of human plague and being reverse result in plague preventionactivities.Materials and MethodsStudy was used data of rodent for zoonotic diseases suspicious which tested plague in National centerfor zoonotic disease (NCZD) in 2005-2015 and 13 local center for zoonotic diseases in 1988-2015. Datawas kept in NCZD and National archival authority. For mapping we used Arc View 3.2.ResultsTotally 397 event information of suspicious rodents and other animals was received in NCZD from 8 districtsof Ulaanbaatar city in 2002-2015. Most of information was received from Songinokhairkhan-64.2%district and smallest number was from Nalaikh district-0.3%. 92.2% of them were marmot, 0.1% of themwere marmot raw products for treatment purpose. Totally 1285 animal samples were tested by plaguedisease and the result was negative. Five hundred thirty tree marmots were carried to Ulaanbaatar from10 provinces. In that time plague foci were active and Y.pestis was isolated in provinces which marmotwas carried to Ulaanbaatar.In 1988-2015, totally 257 marmots and animals of 515 event information was received in15 provinces.Including 13.2% of them were birds, 84% of them marmot, 1.6% of them were livestock, 1.2% of themother animals. About 216 marmots were tested by plague. 51.2% of them were detected positive results.We develop conclusion based laboratory investigation result even it need high cost to take earlyprevention and response measures.Conclusion1. It is high risk to spread plague by carrying suspicious animal in urban area. Therefore, it is importantto take early response measures even it high cost. In further, increase cost and support rapid test ofhigh technology.2. To organize rational advertisement and increase knowledge of population about not doing illegalhunting, not selling marmot raw products in urban area, not using marmot raw products for treatmentuse and avoid contact with marmot carcasses.3. It is important to cooperate joint response measures with policeman, inspection agency andveterinary and human health sectors in Mongolia.