Integrating Psychiatry and Medical Biotechnology as a Way to Achieve Scientific, Precision, and Personalized Psychiatry

en 34900142 Integrating Psychiatry and Medical Biotechnology as a Way to Achieve Scientific, Precision, and Personalized Psychiatry Besides concerns about the increasing prevalence of psychiatric disorders and the significant burdens and costs, there are concerns about its validity. The dilemma of validity went so far that studies described the diagnoses in psychiatry as scientifically worthless. We suggest integrating psychiatry and medical biotechnology and using biotechnological products in psychiatric aspects help psychiatry become more precise, strengthen its position among other sciences, and increase its scientific credibility by giving examples. For this matter, we need different inputs to choose between the vast outputs. The most common inputs are clinical symptoms, cognitive function, individual and environmental risk factors, molecular markers, genetic markers, neuroimaging signs, and big data. Some molecular markers have been shown to have a relationship with psychiatric disorders such as Interleukin-6 (IL-6) and Tumor Necrosis Factor-α (TNF-α). Genetic studies might evolve the most accurate part of precision psychiatry. Currently, and through the developments in technology, genome-wide association studies have become available. In neuroimaging signs, psychiatric disorders are associated with generalized rather than focal brain network dysfunction, and functional magnetic resonance imaging could be performed to study them. It would exhibit different aberrancies in various psychiatric disorders. In big data, the constitution of predictive models and movement toward precision psychiatry can be led by using artificial intelligence and machine learning. Behavioral sciences, Cytokines, Integrative medicine, Knowledge, Technology 172 175 https://www.ajmb.org/En/Article.aspx?id=40485 https://www.ajmb.org/PDF/En/FullText/40485.pdf AhmadShamabadiPsychiatric Research Center, Roozbeh Psychiatric Hospital, Tehran University of Medical Sciences, Tehran, Iran71806AlirezaHasanzadehPsychiatric Research Center, Roozbeh Psychiatric Hospital, Tehran University of Medical Sciences, Tehran, Iran71807ShahinAkhondzadeh739

en 34900143 Apoptosis Induction with Combined Use of Cisplatin and Fisetin in Cisplatin-resistance A2780 Ovarian Cancer Cells Background: Ovarian cancer is the leading cause of death caused by genital cancers. One of the most common treatments for this type of cancer is chemotherapy by cisplatin, which induces apoptosis in cancer cells. Apoptosis is a type of physiological cell death. Cisplatin chemotherapy usually has several side effects and cellular resistance to cisplatin is a common incidence. In order to overcome these problems, the use of combination therapies using natural substances has been considered. Fisetin is a flavonoid with anti-cancer activity which induces apoptosis. In this study, the apoptosis induced by cisplatin along with Fisetin in cisplatin-resistant ovarian cancer cell line (A2780) was investigated. Methods: In the present experimental study, the effect of combined use of Fisetin and cisplatin on ovarian cancer cell lines (A2780) was investigated by using MTT assay. Cell death was also determined by DAPI, acridine orange/propidium iodide, and Annexin/PI assay. Apoptotic gene expression of Bax, BCL-2, caspase 3, and caspase 9 was also assessed by real time PCR. Results: The results of MTT assay indicated that the combined treatment of Fisetin and cisplatin effectively inhibits proliferation of A2780 cells. The results of DAPI staining showed that fragmentation of chromatin in cells occurred in the combined treatment. Acridine orange-propidium iodide staining and Annexin/PI staining showed an increase in the rate of apoptotic cells in cells under combined treatment. The results of the study regarding changes in gene expression also indicated that Bax pro-apoptotic gene expression and BCL-2 anti-apoptotic gene expression increased in cells under treatment; moreover, gene expression of caspases 3 and 9 significantly increased as well. Conclusion: According to the findings of this study, the combined use of cisplatin and Fisetin increases the induction of apoptosis in cisplatin-resistant ovarian cancer cells (A2780); therefore, the combined use of cisplatin and Fisetin can be considered a promising strategy in the treatment of ovarian cancer. Apoptosis, Cisplatin, Fisetin, Ovarian neoplasms 176 182 https://www.ajmb.org/En/Article.aspx?id=40474 https://www.ajmb.org/PDF/En/FullText/40474.pdf SamiraJafarzadehDepartment of Biology, Mashhad branch, Islamic Azad University, Mashhad, Iran71803JavadBahararaResearch Center for Animal Development Applied of Biology, Mashhad branch, Islamic Azad University, Mashhad, Iran71804MaryamTehranipourDepartment of Biology, Mashhad branch, Islamic Azad University, Mashhad, Iran71805

en 34900144 A Cross-Sectional Study for Evaluation of KRAS and BRAF Mutations by Reverse Dot Blot, PCR-RFLP, and Allele-Specific PCR Methods Among Patients with Colorectal Cancer Background: KRAS and BRAF genes are the biomarkers in Colorectal Cancer (CRC) which play prognostic and predictive roles in CRC treatment. Nowadays, the selection of rapid and available methods for studying KRAS and BRAF mutations in anti-EGFR therapy of patients suffering from CRC plays a significant role. In this study, the mutations of these two oncogenes were evaluated by different methods. Methods: This study was performed on 50 Formalin-Fixed Paraffin-Embedded (FFPE) tissue blocks of patients diagnosed with colorectal cancer. After DNA extraction, KRAS and BRAF gene mutations were evaluated using reverse dot blot, and results were compared with PCR-RFLP and allele-specific PCR for KRAS and BRAF mutations, respectively. Results: KRAS gene mutations were detected in 42% of patients, of which 30% were in codon 12 region, and 12% in codon 13. The most frequent mutations of KRAS were related to G12D and 10% of patients had BRAF mutated genes. The type of KRAS gene mutations could be evaluated by reverse dot blot method. In general, the results of PCR-RFLP and allele-specific PCR were similar to the findings by reverse dot blot method.  Conclusion: These findings suggest that PCR-RFLP and allele-specific PCR methods are suitable for screening the presence of the mutations in KRAS and BRAF oncogenes. In fact, another method with more sensitivity is needed for a more accurate assessment to determine the type of mutations. Due to higher speed of detection, reduced Turnaround Time (TAT), and possible role of some KRAS point mutations in overall survival, reverse dot blot analysis seems to be an optimal method. Allele-Specific PCR, BRAF, Colorectal neoplasms, KRAS, PCR-RFLP, Reverse dot blot 183 191 https://www.ajmb.org/En/Article.aspx?id=40475 https://www.ajmb.org/PDF/En/FullText/40475.pdf FatemehSheikhsoflaDepartment of Cellular and Molecular Biology, University of Mazandaran, Mazandaran, Iran71808BehzadPoopak71809SajjadFiruzyarRazi Vaccine and Serum Research Institute of Karaj, Karaj, Iran71810FatemehRoudbari Department of Virology, University of Mazandaran, Mazandaran, Iran71811MojtabaGhadianyDepartment of Hematology and Oncology, Shahid Beheshti University of Medical Sciences, Tehran, Iran71812

en 34900145 Production of PEGylated G-CSF from non-classical inclusion bodies expressed in Escherichia coli Background: The recombinant human granulocyte colony stimulating factor conjugated with polyethylene glycol (PEGylated GCSF) has currently been used as an efficient drug for the treatment of neutropenia caused by chemotherapy due to its long circulating half-life. Previous studies showed that Granulocyte Colony Stimulating Factor (GCSF) could be expressed as non-classical Inclusion Bodies (ncIBs), which contained likely correctly folded GCSF inside at low temperature. Therefore, in this study, a simple process was developed to produce PEGylated GCSF from ncIBs. Methods: BL21 (DE3)/pET-GCSF cells were cultured in the LiFlus GX 1.5 L bioreactor and the expression of GCSF was induced by adding 0.5 mM IPTG. After 24 hr of fermentation, cells were collected, resuspended, and disrupted. The insoluble fraction was obtained from cell lysates and dissolved in 0.1% N-lauroylsarcosine solution. The presence and structure of dissolved GCSF were verified using SDS-PAGE, Native-PAGE, and RP-HPLC analyses. The dissolved GCSF was directly used for the conjugation with 5 kDa PEG. The PEGylated GCSF was purified using two purification steps, including anion exchange chromatography and gel filtration chromatography. Results: PEGylated GCSF was obtained with high purity (~97%) and was finally demonstrated as a form containing one GCSF molecule and one 5 kDa PEG molecule (monoPEG-GCSF). Conclusion: These results clearly indicate that the process developed in this study might be a potential and practical approach to produce PEGylated GCSF from ncIBs expressed in Escherichia coli (E. coli). Granulocyte colony stimulating factor (G-CSF), Inclusion bodies, Polyethylene glycol 192 200 https://www.ajmb.org/En/Article.aspx?id=40477 https://www.ajmb.org/PDF/En/FullText/40477.pdf NguyenThi My TrinhVietnam National University, Ho Chi Minh City, Vietnam71800TranLinh ThuocVietnam National University, Ho Chi Minh City, Vietnam71801DangThi Phuong ThaoDepartment of Molecular and Environmental Biotechnology, University of Science, Ho Chi Minh City, Vietnam71802

en 34900146 Fabrication of Calcium Sulfate Coated Selenium Nanoparticles and Corresponding In-Vitro Cytotoxicity Effects Against 4T1 Breast Cancer Cell Line Background: The inhibitory effect of selenium nanoparticles (SeNPs) on cancer cells has been reported in many studies. In this study,  the purpose was to compare the in vitro effects of SeNPs and calcium sulfate coated selenium nanoparticles (CaSO4@ SeNPs) on breast cancer cells. Methods: CaSO4@SeNPs and SeNPs were chemically synthesized and characterized with Field Emission Scanning Electron Microscope (FESEM) and energy-dispersive X-ray spectroscopy (EDX). By applying MTT assay, the cytotoxicity effect of both nanomaterials on the 4T1 cancer cells was investigated. Results: While LD50 of SeNPs on 4T1 cancer cells was 80 µg, the LD50 of CaSO4@SeNPs was reported to be only 15 µg. The difference between the inhibition rates obtained for SeNPs and CaSO4@SeNPs was statistically significant (p=0.05). In addition, at higher concentrations (50 µg) of CaSO4@SeNPs, the cytotoxicity was 100% more than SeNPs alone.  Conclusion: According to the result of the present work, it can be concluded that decoration of SeNPs with calcium sulfate leads to an increase in potency by decreasing the effective dose. This effect can be attributed to activation of intrinsic apoptosis signaling and/or pH regulatory properties of CaSO4@SeNPs. However, further studies are still needed to determine the exact corresponding mechanisms of this synergistic effect. Apoptosis, Breast neoplasms, Calcium sulfate, Nanoparticles, Selenium 201 206 https://www.ajmb.org/En/Article.aspx?id=40478 https://www.ajmb.org/PDF/En/FullText/40478.pdf ElnazFaghfuri Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran71777RamakAjidehBiotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran71778FaranakShahverdi Recombinant Vaccine Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran71779MinaHosseiniDepartment of Pharmaceutical Biotechnology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran71780FaranakMavandadnejadDepartment of Pharmaceutical Biotechnology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran71781Mohammad HosseinYazdiDepartment of Pharmaceutical Biotechnology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran534Ahmad RezaShahverdiRecombinant Vaccine Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran59

en 34900147 Molecular Mechanisms of Anti-inflammatory Activities of the Extracts of Ocimum gratissimum and Thymus vulgaris Background: A large body of literature suggests that the extracts of Ocimum gratissimum (O. gratissimum) and Thymus vulgaris (T. vulgaris) play protective roles against various inflammatory disorders. However, the possible mechanism of action with reference to the interactions of their respective phytochemical compositions with pro-inflammatory mediators as the indication of their therapeutic effects is less clear. Therefore, the immunomodulatory properties of O. gratissimum and T. vulgaris were investigated in this study. Methods: The in vitro lipoxygenase inhibitory potentials of methanolic extracts of the selected plants were assessed through colorimetric analysis. The pharmacokinetics of some identified compounds in the botanicals were investigated via the Swiss ADME server while the molecular interactions of the compounds with lipoxygenase, IL-1, IL-6, TNF-α, IL-8, and CCL-2 were performed through molecular docking. Results: The assessment of the lipoxygenase inhibition revealed the extracts could possess anti-inflammatory agents. The pharmacokinetic results of some selected compounds identified in the botanicals showed moderate toxic effects compared to indomethacin. The molecular docking study substantiated the report of the in vitro analysis as indicated in the binding score of all the selected compounds compared to indomethacin. Conclusion: The phytochemical components of the extracts of O. gratissimum and T. vulgaris could be effective as anti-inflammatory agents that could be explored in preventing disorders associated with excessive activities of pro-inflammatory mediators. Anti-inflammatory agents, Lipoxygenase, Ocimum, Phytochemicals, Plant extracts, Thymus plant 207 216 https://www.ajmb.org/En/Article.aspx?id=40479 https://www.ajmb.org/PDF/En/FullText/40479.pdf IgeFrancis OlaoyeDepartment of Biochemistry, McPherson University, Seriki Sotayo, Ogun State, Nigeria71797BabatundeJoseph Oso71798AdepejuAberuagbaDepartment of Biochemistry, University of Ilorin, Ilorin, Kwara State, Nigeria71799

en 34900148 Construction and Evaluation of Short Hairpin RNAs for Knockdown of Metadherin mRNA Background: Short hairpin RNA (shRNA) has proven to be a powerful tool to study genes’ function through RNA interference mechanism. Three different methods have been used in previous studies to produce shRNA expression vectors including oligonucleotide-based cloning, polymerase chain reaction (PCR)-based cloning, and primer extension PCR approaches. The aim of this study was designing a reliable and simple method according to the primer extension strategy for constructing four shRNA vectors in order to target different regions of Metadherin (MTDH) mRNA in human leukemic cell line Jurkat. Methods: Oligonucleotides for construction of four shRNA vectors were designed, synthesized and fused to U6 promoter. Each U6-shRNA cassette was cloned into a pGFP-V-RS vector. MTDH shRNAs were transfected into the Jurkat cell line by using the electroporation method. The ability of shRNAs to knock down MTDH mRNA was analyzed through qRT-PCR. Apoptosis assay was used to evaluate the effect of down regulation of MTDH expression on cell integrity. Results: A significant reduction (about 80%) in the expression levels of MTDH mRNA and an increase in the percentages of apoptotic cells (about 20%) were observed in the test group in comparison with control. Conclusion: MTDH shRNA constructs effectively inhibited gene expression. However, simplicity and inexpensiveness of the method were additional advantages for its application. Apoptosis, Gene expression, Gene silencing, Jurkat cells, RNA 217 222 https://www.ajmb.org/En/Article.aspx?id=40480 https://www.ajmb.org/PDF/En/FullText/40480.pdf FarahnazZareDiagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran71786SedighehSharifzadeh 71787AbbasBehzad-BehbahaniDiagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran899GholamrezaRafiei DehbidiDiagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran71788ZahraYousefiSchool of Allied Medical Sciences, Shahroud University of Medical Sciences, Shahroud, Iran71789RezaRanjbaranDiagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran898NoorossadatSeyyediDiagnostic Laboratory Sciences and Technology Research Center, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran1349

en 34900149 Evaluation of miR-222 Expression in HBV Infected Patients in Comparison with HDV and HBV Co-infected Patients Background: Liver disease is more severe in HDV+HBV co-infected patients than HBV infected patients which seems to be related to differences in the expression of genes and other factors such as MicroRNAs (miRNAs). The aim of this study was to investigate miR-222 expression in HBV infected patients in comparison with HDV+HBV co-infected patients. Methods: First, total RNA was extracted from the serum samples and then, complementary DNA (cDNA) was produced using cDNA synthesis kit. Finally, miR-222 gene expression was measured using U6 as the internal control by quantitative PCR (qPCR). Results: The level of miR-222 expression in HDV+HBV co-infected samples was significantly up regulated. The fold change of the miR-222 expression between two groups was 3.3 (95% CI; 0.011- 17.63) with p<0.001. Conclusion: The expression of miR-222 was higher in HBV+HDV co-infected patients than HBV infected patients. Further studies should be conducted to confirm whether miR-222 can be a biomarker for prognosis of severe liver diseases. Co-infection with hepatitis B and D, Hepatitis B virus, Hepatitis D virus, MiR-222 223 225 https://www.ajmb.org/En/Article.aspx?id=40481 https://www.ajmb.org/PDF/En/FullText/40481.pdf ZahraSokhanvarDepartment of Microbiology, Faculty of Biological Sciences, Alzahra University, Tehran, Iran71783AmenehElikaei Department of Microbiology, Faculty of Biological Sciences, Alzahra University, Tehran, Iran71785ZohrehSharifi640

en 34900150 Pitfalls of Restriction Enzyme Mapping Following Generation of CRISPR Constructs Background: The PX330 and the related PX459 plasmids are widely used for Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)/Cas9-mediated genome editing. Screening for plasmids containing the correct sgRNA template insertion is one of the most important steps in this system. Different methods for screening the sgRNA inserts have been deployed. One such method is Restriction Enzyme (RE) mapping. Restriction enzyme mapping can be used to screen for numerous plasmid recombinants simultaneously. Methods: In this study, the sgRNA templates were initially cloned into the above PX459 plasmids. Subsequently, the accuracy of the constructs was determined by RE mapping. Results: This method was established to screen for sgRNA-bearing PX459 plasmids. However, numerous anomalies were detected after ligation of sgRNA templates into RE digested PX459 plasmids. Conclusion: Our data suggest that RE mapping is only appropriate as an initial screen and that the identity of all plasmids with the correctly identified RE maps should be confirmed by Sanger sequencing. CRISPR-Cas systems, Gene editing, Health services, Plasmids, Restriction mapping 226 229 https://www.ajmb.org/En/Article.aspx?id=40483 https://www.ajmb.org/PDF/En/FullText/40483.pdf MehdiHassaniGenetics Research Center, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran71776SaraHesamiGenetics Research Center, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran71774NahalMaroofiGenetics Research Center, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran71775MehdiBanan1152

en 34900151 Identification of a Novel Homozygous Mutation in BBS10 Gene in an Iranian Family with Bardet-Biedl Syndrome Background: Bardet–Biedl Syndrome (BBS) is a rare pleiotropic autosomal recessive disease related to ciliopathies with approximately 25 causative genes. BBS is a multisystemic disorder with wide spectrum of manifestations including truncal obesity, retinal dystrophy, male hypogenitalism, postaxial polydactyly, learning difﬁculties, and renal abnormalities. Methods: A consanguineous Iranian family with a 28-year-old daughter affected with BBS, resulting from a first cousin marriage, was examined. After clinical examination, Whole Exome Sequencing (WES) was applied. Following the analysis of exome data, Sanger sequencing was used to confirm as well as to co-segregate the candidate variant with the phenotype. Results: A novel homozygous variant [c. 2035G>A (p.E679K)] in exon 2 of the BBS10 gene was found which was categorized as likely pathogenic based on American College of Medical Genetics and Genomics (ACMG) guidelines and criteria. In this study, the variant was fully co-segregated with the phenotype in the family. Conclusion: Despite overlapping with other ciliopathies in terms of the phenotype, the BBS has high genetic heterogeneity and clinical variability even among affected members of a family. The symptoms observed in patients are largely related to the genes involved and the type of mutations in the BBS. In this study, in addition to phenotype description of the proband harboring a novel disease-causing variant in BBS10 gene, the spectrum of BBS symptoms was expanded. The findings of this study can be useful in genetic counseling, especially for risk estimation and prenatal diagnosis. Bardet–Biedl syndrome, Mutation, Whole exome sequencing 230 233 https://www.ajmb.org/En/Article.aspx?id=40482 https://www.ajmb.org/PDF/En/FullText/40482.pdf MohammadDehaniGenetics Research Center, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran71792DavoodZare-Abdollahi11425AtaBushehri Genetics Research Center, University of Social Welfare and Rehabilitation Sciences, Tehran, Iran71793AzadehDehghani Medical Biotechnology Research Center, Ashkezar Branch, Islamic Azad University, Ashkezar, Iran71794JalilEffatiMeybod Genetic Research Center, State Welfare Organization, Yazd, Iran71795Seyed Ali MohammadMiratashi Department of Ophthalmology, Shahid Sadoughi University of Medical Sciences, Yazd, Iran71796Hamid RezaKhorram KhorshidFetal Health Research Center, Hope Generation Foundation, Tehran, Iran42

en 34900152 The Potential Anti-inflammatory Effects of Zerumbone in COVID-19 Patients 234 236 https://www.ajmb.org/En/Article.aspx?id=40484 https://www.ajmb.org/PDF/En/FullText/40484.pdf RaziehDehghanResearch Center for Molecular Medicine, Faculty of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran71769Mohammad HasanSoheilifarDepartment of Medical Laser, Medical Laser Research Center, Yara Institute, ACECR, Tehran, Iran1325FaridAzizi JalilianDepartment of Virology, Faculty of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran71771RezvanNajafiResearch Center for Molecular Medicine, Faculty of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran71772RaziehAmini71773