Construction of pLLO vector encoding truncated form of Listeriolysin O as molecular adjuvant for DNA vaccine studies

Background: The major problem of DNA vaccine is less immunogenicity of them verses other killed or live whole organism vaccines therefore adjuvants for use in this kind vaccines is very necessary. Genetic adjuvants with bacterial sources are an appropriate approach to modulate immune responses to DNA vaccines. Listeria Monocytogenes proteins such as Listeriolysin O (LLO) with CD4 and CD8 epitopes can be as an adjuvant to initiate both innate and adaptive immune responses if the protein cytotoxicity can be eliminated. Herein we constructed a truncated LLO plasmid as genetic adjuvant and tested it in combination with a DNA construct as a model vaccine.Materials and Methods: About 1340bp of the 5' end of whole LLO gene was amplified by PCR on DNA purified from Listeria Monocytogenes. Sequential sub cloning of truncated LLO into the Xho I/EcoRV sites of pcDNA3.1 plasmid, downstream of CMV promoter was done. pLLO plasmid was transfected to HEK293T cell line by lipofection method. LLO protein expression from transiently transfected 293T cell lysates was confirmed by western blotting. Then the adjuvant activity of LLO in BALB/c mice model was analyzed using proliferation test.Results: Double digestion of pLLO plasmid with the enzymes that were applied for cloning led to the isolation of two fragments with expected sizes. The final plasmid was also confirmed following sequencing reactions. Moreover, expression of LLO was evidenced in transfected 293T cells, compared to non-transfected controls. In vivo study was shown, high significant proliferative responses in LLO co-immunization pattern.Conclusion: In the DNA vaccine study, LLO co-administration plasmid could be a suitable genetic adjuvant to enhance cellular immune response of vaccine

Detection of human CMV PP65 protein in glioma brain tumors with immunohistochemistry method

Background: Human cytomegalovirus (HCMV) may play a role in the development of glioma disease that is one of the most common brain tumors. Objective: The aim of this study was to detect human CMV in patients with glioma in Imam Khomeini hospital, Tehran. Methods: This experimental study was conducted on paraffin-embedded tumor samples of 18 patients referred to Imam Khomeini hospital in 2012. Immunohistochemistry (IHC) was performed with monoclonal antibody specific for HCMV PP65 protein and the samples were assessed using a light microscope. Findings: Of 18 patients, 13 (72.2%) were positive for HCMV PP65 protein and four of them expired. Conclusion: With regards to the results, more comprehensive studies are recommended for detection of HCMV in patients with glioma using different diagnostic methods. Keywords: Cytomegalovirus, Immunohistochemistry, Gliom

Comparative Effects of Parent Management Training Combined With ACT and Mindful Parenting on Parent-child Relationship

Background: Considering the profound influence of parent-child relationships on child vulnerability and mental health, prioritizing the improvement of this bond becomes crucial. While combined programs have garnered substantial recognition for their heightened effectiveness, regrettably, no research has yet explored a comparative analysis of their efficacy. This study aims to assess and compare the effectiveness of combining parent management training (PMT) with both acceptance commitment therapy (ACT) and mindful parenting in improving the parent-child relationship. Methods: The study employed a quasi-experimental design, specifically a pre-test and post-test method, with a three-month follow-up period. A control group was included to enhance the validity of the results. The statistical population comprised all mothers of preschool children in Semnan City, Iran in 2022. Using convenience sampling, 36 mothers were selected and randomly assigned to three groups, two experimental groups and one control group. Random assignment was achieved using a lottery-based method without replacement. The sample in experimental groups participated in an integrated training program, which consisted of two-hour sessions conducted eight times per week. Conversely, the control group was placed on a waiting list. The data analysis phase involved utilizing SPSS software, version 23 and applying multivariate covariance analysis.  Results: The results of the study indicated that both combined programs exhibited a significant impact on reducing conflicts within the parent-child relationship and fostering closeness (P0.05). Additionally, no significant difference was observed between the two combined programs regarding their effectiveness in enhancing the parent-child relationship. These results remained consistent during the follow-up stage.  Conclusion: This study highlighted that these programs have exhibited a significant capacity to enhance the parent-child relationship by fostering a sense of closeness and effectively mitigating conflicts. These results indicated that the integration of ACT or mindful parenting principles with PMT can provide valuable interventions for enhancing parent-child

Determining the frequency of reporting laboratory data in summary sheets of patients in a pediatric ward based on the classification of diagnoses in ICD-10

Background: A summary of clinical findings and laboratory data is collected in patient summary sheets. The aim of this study was to determine the frequency of laboratory data reported in the summary sheets based on the classification of diagnoses in ICD-10. Methods: A descriptive-cross sectional study was performed on summary sheets of patients admitted to the pediatric ward from March 2015 to February 2016.The laboratory data of these sheets were extracted using a data collection form. Because of high number of data in each sheet and given that, depending on the diagnosis, the reported laboratory data were different, to simplify the presentation of the results the data were categorized based on patients’ diagnoses which were coded by ICD-10. Results: A total of 10224 laboratory data were extracted, which included 47 unique data elements. The highest number of laboratory data was related to hematology (N = 14) and the lowest number was related to urine and stool analysis (N = 6). Conclusions: Laboratory data are recorded in summary sheets in accordance with patient's diagnosis. Therefore, it is possible to process the existing summary sheets and extract the registration rules, then implement these rules in hospital information systems to generate electronic summary sheets

Construction and Eukaryotic Expression of Recombinant Large Hepatitis Delta Antigen

Background: Hepatitis delta virus (HDV) is a subviral human pathogen that exploits host RNA editing activity to produce two essential forms of the sole viral protein, hepatitis delta antigen (HDAg). Editing at the amber/W site of HDV antigenomic RNA leads to the production of the large form (L-HDAg), which is required for RNA packaging. Methods: In this study, PCR-based site-directed mutagenesis by the overlap extension method was used to create the point mutation converting the small-HDAg (S-HDAg) stop codon to a tryptophan codon through three stages. Results: Sequencing confirmed the desirable mutation and integrity of the L-HDAg open reading frame. The amplicon was ligated into pcDNA3.1 and transfected to Huh7 and HEK 293 cell lines. Western blot analysis using enhanced chemiluminescence confirmed L-HDAg expression. The recombinant L-HDAg localized within the nuclei of cells as determined by immunofluorescence and confocal microscopy. Conclusion: Because L-HDAg requires extensive post-translational modifications, the recombinant protein expressed in a mammalian system might be fully functional and applicable as a tool in HDV molecular studies, as well as in future vaccine research

A DNA Vaccine-Encoded Nucleoprotein of Influenza Virus Fails To Induce Cellular Immune Responses in a Diabetic Mouse Model▿

Influenza virus infections cause yearly epidemics and are a major cause of lower respiratory tract illnesses in humans worldwide. Influenza virus has long been recognized to be associated with higher morbidity and mortality in diabetic patients. Vaccination is an effective tool to prevent influenza virus infection in this group of patients. Vaccines employing recombinant-DNA technologies are an alternative to inactivated virus and live attenuated virus vaccines. Internal highly conserved viral nucleoprotein (NP) can be delivered as a DNA vaccine to provide heterosubtypic immunity, offering resistance against various influenza virus strains. In this study, we investigated the efficacy of an NP DNA vaccine for induction of cell-mediated immune responses and protection against influenza virus infection in a mouse model of diabetes. Healthy and diabetic BALB/c mice were immunized on days 0, 14, and 28 by injection of NP DNA vaccine. Two weeks after the last immunization, the cellular immune response was evaluated by gamma interferon (IFN-γ), lymphocyte proliferation, and cytotoxicity assays. The mice were challenged with influenza virus, and the viral titers in the lungs were measured on day 4. Diabetic mice showed significantly smaller amounts of IFN-γ production, lymphocyte proliferation, and cytotoxicity responses than nondiabetic mice. Furthermore, higher titers of the influenza virus were detected after challenge in the lungs of the diabetic mice. The present data suggest that the NP DNA vaccine with the protocol of immunization described here is not able to induce efficient cellular immune responses against influenza virus infection in diabetic mice

The concept and forms of yang sheng in Chinese society: socio-philosophical analysis

Актуальность исследования обусловлена происходящими трансформациями традиционной китайской медицины в современном китайском обществе. Практики ян шэн (практики «вскармливания жизни») занимают важное место в традиционной китайской медицине, имеют многовековую историю и нацелены на поддержание гармоничного состояния организма. Однако под влиянием западной медицины и культуры возникают новые формы ян шэн и практики здоровьесбережения современных жителей Китая находятся в процессе адаптации к новым изменениям. Ключевой задачей данной статьи является социально-философский анализ понятия ян шэн в китайском обществе. В статье будут рассмотрены социально-философские основания концепции ян шэн и, в контексте традиционной китайской медицины, проанализирован ряд ключевых практик «вскармливания жизни» - в китайском обществе. Методы: изучение и анализ литературных источников с акцентом на историко-философский и социально-культурный анализ темы исследования. Выводы. Было детально рассмотрено понятие историко-культурного феномена ян шэн в современном китайском обществе, его связь с традиционной китайской медициной, а также традиционные и современные формы практик «вскармливания жизни».The relevance of the study is caused by transformations of traditional Chinese medicine occurring in modern Chinese society. Practiсes of yang sheng (the practices of «life nurturing») occupy an important place in traditional Chinese medicine, have a long history and are aimed at maintaining a harmonious state of the body. However, under the influence of Western medicine and culture, new forms of yang sheng are emerging, and the practices of health preservation of modern Chinese people is in adapting to new changes. The key objective of the article is a socio-philosophical analysis of the concepts of yang sheng in Chinese society. The paper will examine the socio-philosophical foundations of the concept of yang sheng and analyze a number of key practices of «life nurturing» in Chinese society in the context of traditional Chinese medicine. Methods: study and analysis of literary sources with an emphasis on historico-philosophical and socio-cultural analysis of the research topic. Results. The authors reviewed in detail the concept of the historico-cultural phenomenon of yang sheng in modern Chinese society, its connection with traditional Chinese medicine, as well as traditional and modern forms of the practices of «life nurturing»