فصلنامه تازه های بیوتکنولوژی سلولی مولکولی
پیاپی 43 (تابستان 1400)

Coronaviruses are a large family of viruses that cause a variety of ailments ranging from the common cold to acute and severe respiratory syndrome. The emergence of Covid 19 has many psychological effects, one of which is anxiety and has physical and psychological effects. However, most studies have focused only on clinical data. The virus has now spread to all countries and has become an epidemic, rapidly endangering mental and physical health. Therefore, the aim of the present study was to investigate the correlation between anxiety and the prevalence of Covid 19.
In this review article, the psychological effects of COVID-19 and its relationship with anxiety were investigated. Accordingly, using electronic resources in reputable databases of PubMedScopus, SID, Google Scholar, Science Direct, indexed articles were obtained from 114 articles, 35 articles were evaluated and 13 articles entered the secondary screening stage.
The present study showed that COVID-19 has psychological problems other than mortality. Studies in other parts of the world have shown that COVID-19 has several psychological effects, including increased anxiety.
Due to its unknown nature, it causes anxiety in affected individuals and other members of the community. With increasing prevalence of COVID-19 and its limitations, the level of anxiety also increases. Therefore, increasing public awareness of this disease and presenting positive psychological programs in the media aimed at controlling stress can reduce anxiety in society.

Candida species are opportunistic pathogens that can be considered as the main cause of a wide range of diseases. Due to the increased resistance of these fungi to chemical drugs and also the adverse effects of these compounds on patients, it is necessary to replace these drugs with herbal compounds. In this study, the antifungal effect of aqueous and ethanolic extracts and essential oil of E. angustifolia on the growth of Candida albicans with fluconazole was compared.

In this laboratory study conducted in 1398 in Shahrekord Azad University Biotechnology Research Center, 80 candidate fungal samples were prepared from Parsian Hospital in Shahrekord and 50 Candida albicans samples were isolated by diagnostic tests. E. Angustifolia plants were obtained from Shahrekord Azad University Faculty of Agriculture. The extract was extracted by percolation and the essential oil was extracted by water distillation. The antifungal effect of E. angustifolia extracts along with E. angustifolia essential oil with fluconazole was compared on these isolates by disk diffusion and microdilution methods.

The minimum and maximum diameter zone of inhibation obtained by disk diffusion method for E. angustifolia extracts, no growth halo formation with 1 and the lowest and maximum halo diameter for E. angustifolia essential oil were 6 and 37 mm. Therefore, the mean diameter of non-growth zone of inhibation for E. angustifolia extracts was 1 and for E. angustifoliathe essential oil was 23.4 mm. Comparison of the mean diameter of 25 mg fluconazole inhibition zone as control with an average of 16.26 mm shows a weaker antifungal performance and a significant difference compared to E. angustifolia essential oil. (P ≤ 0.05). The minimum inhibitory concentrations (MIC) of ethanolic and aqueous extracts of E. angustifolia and fluconazole were 4, 8, and 8 mg/ml, respectively; While the minimum fungicidal concentrations(MFC) were 8, 16, and 8 mg/ml, respectively; Showed a significant difference in inhibition of E. angustifoli ethanolic extract compared to 25 mg fluconazole disk (P ≤ 0.01).

According to the results of the above research, it seems that the ethanolic extract of E. angustifoliacan be used as a suitable drug candidate with better antifungal performance than fluconazole in the pharmaceutical industry.

Today, many patients with coagulation disorders due to defective or non-functioning coagulation factors and the use of various blood thinners need to check their prothrombin total and quantify determination of direct thrombin inhibitors. Despite the study of the ecarin from Echis carinatus venom as a direct activator of prothrombin, there have been not investigations into the active site of this enzyme.

Ecarin metalloproteinase domain (639 bp) was synthesized into the pcDNA3.1. Recombinant plasmid was transfected into HEK293 cell line. Protein expression was evaluated using SDS-PAGE and its function in reaction with prothrombin and using a prothrombin time test was measured.

The results showed that the metalloproteinase domain of ecarin produced in the HEK293, like whole ecarin, was able to activate prothrombin to thrombin; but its activity was slow than one produced in the prokaryotic system.

In this study, the recombinant ecarin metalloproteinase produced in HEK293 cells, like whole ecarin, was able to activate prothrombin to thrombin and was introduced for the first time as a suitable alternative to both natural and recombinant ecarin in methods based on prothrombin to thrombin conversion.

Cancer is a major cause of mortality in the world. Treatment of Cancer through drug therapy/chemotherapy faces the challenges of drug delivery. Drug delivery systems such as niosomes provided high efficiency and targeted treatment. The aim of this study was to prepare a new formula of niosomal nano system containing doxorubicin as the cancer therapy drug. Changing the tissue distribution of drug leads to drug effect improvement and reducing its cytotoxicity.

Doxorubicin encapsulated niosomes were synthesized using thin-film method. A certain amount of Span-60 and Cholesterol dissolved by chloroform in a round bottom balloon. Rotary evaporator was used in order to remove the organic solvent and form the thin-film. The thin-film was hydrated by doxorubicin and PBS solution using rotary evaporator in 10°C higher than span-60 transition phase. Entrapment Efficiency and release profile were investigated using dialysis and spectrophotometry. Size reduction was applied by probe sonicating. Nanoparticles average diameter and zeta potentiol was measured using DLS technique and Zeta sizer. The optimum formula was PEGylated and its cytotoxicity investigated through MTT assay on Ovarian cancer cell line and fibroblast normal cells.

The diameter of optimized and PEGylated niosomal formulation was 172.5 nm and entrapment efficiency of PEGylated doxorubicin system was about 95.83±1.18. release studies result in PBS buffer at 37°C was the sign of PEGylated nanoniosomes efficiency in controlling the drug release, which the release percent of drug after 48 hours was 20.52 and its cytotoxicity was higher than the free drug.

The study demonstrates using of drug delivery carriers such as synthesized nano-niosomes has an effective role in increasing the efficacy and reducing the consumed dosages of drug.

Coronary arteries are blood vessels that carry blood to the heart. With plaque forming on the walls of these arteries, the blood supply becomes problematic. The diagnosis of coronary artery disease is very important because of the risks to personchr('39')s health. The aim of this study is to determine genes with differential expression between CAD patients and normal individuals in the ErbB signaling pathway, as one of the important pathways in the development of this disease, and to investigate their importance in identifying biomarkers of coronary artery disease.

The raw files of RNA-sequencing of samples of coronary artery patients and normal individuals were obtained from datasets with numbers GSE99985 and GSE100206, respectively, and the steps of sample analysis were performed. Genes with different expression in R program were isolated with DESeq2 package. Then, functional analysis of genes including signaling pathways, biological processes and genes with different expression in the ErbB pathway and how they change were examined.

The results demonstrated upregulation of 1069 genes and downregulation of 851 genes among samples. In addition, functional analysis revealed that genes such as CAMK2D, SHC3, ERBB3, NRG4, RPS6KB2, MAPK1, BRAF, AREG and CRK had differentially expression among patients could affect ErbB signaling pathway as well.

 Molecular studies can help to achieve the reliable biomarkers of coronary artery disease which is important for biomedical research, drug development and clinical applications.

Human papillomaviruses (HPVs) especially types 16 and 18 are known as the major causes of cervical cancer. Among HPV proteins, L1 and L2 capsid proteins, and also E7 oncoprotein are proposed as target antigens for vaccine design. In the recent years, the recombinant multiepitope polypeptides have attracted a special interest. The goal of this study is the design of L1-L2-E7 fusion construct and evaluation of its expression in bacterial expression system. 

In this study, the immunogenic and conserved epitopes of HPV16/18 L1, L2 and E7 proteins were selected using different bioinformatics analyses. After synthesis of the designed L1-L2-E7 fusion sequence in pUC57 cloning vector, its subcloning was performed in pET24a (+) prokaryotic expression vector using EcoRI/ HindIII restriction enzymes. The expression of the recombinant multiepitope polypeptide was done in E.coli Rosetta strain using IPTG inducer, and confirmed by SDS-PAGE and western blotting using anti-His-tag antibody. The expression was optimized under different conditions such as optical density (OD in wavelength of 600 nm), temperature and time after induction, and IPTG concentration.   

The recombinant pET-L1-L2-E7 vector was confirmed by the presence of a clear band (~525 bp) related to the L1-L2-E7 gene on agarose gel after enzyme digestion. The expression of L1-L2-E7 polypeptide in bacteria showed the presence of a clear band (~20 kDa) in SDS-PAGE and western blotting. The best conditions for expression of the recombinant polypeptide were at temperature of 37◦C, optical density of 0.7-0.8, IPTG concentration of 1mM, and time of 16 h after induction.  

The successful expression of the L1-L2-E7 multiepitope polypeptide was performed in bacterial system. In the next step, the recombinant polypeptide will be purified to use as a vaccine candidate.

Gastric cancer is one of the most common cancers with high mortality. B7-H4 has been identified as potential novel diagnostic biomarker in gastric cancer. But, its clinical significance in this area is still unclear. The purpose of the present study was to examine the expression level of B7-H4 mRNA in tumor tissue of gastric cancer patient with and without history in comparison with their marginal tissue. Also we evaluate its role in tumor progression and prognosis.  

In this study, paraffin blocks were studied from 60 patients with gastric adenocarcinoma (30 with metastasis at the time of diagnosis and 30 without evidence of metastasis) who underwent gastrectomy during 1387-97 in Imam Khomeini Hospital in Tehran. Samples were taken from paraffin blocks by punching method (including tumor and tumor margin for each sample) after the area was identified by the pathologist. RNA extraction was performed from tumor sections and tumor margins. B7-H4 mRNA expression was assessed by quantitative Real-time polymerase chain reaction (qRT-PCR).

Increased expression of B7-H4 mRNA was observed in 81.5% of tumor tissues. There was a significant difference in B7-H4 mRNA expression in tumor tissues of cases with metastasis (p <0.001) and without metastasis (p <0.003) compared to tumor margin. However, comparison of B7-H4 mRNA expression between tumor tissues of metastatic and non-metastatic individuals did not show a significant difference (p = 0.732). Analyzes did not show a significant relationship between B7-H4 mRNA expression and clinical and histopathological variables.

Our findings indicate the possible role of B7-H4 as a potential diagnostic biomarker in patients with gastric cancer. Although more research is needed in this area.

With the advent of cloning technology and recombinant DNA, it became possible to produce drugs and vaccines that were not possible in the past. Recombinant protein occupies a large part of the biopharmaceutical industry, one of the most important of which is vaccination.

In this study, with the aim of obtaining a protein candidate for cholera vaccine, first cloning of a plasmid designed for the bacterial host E.coli DE3 Rossetagami was transformed, then expression was induced using IPTG and after protein expression was examined by SDS-PAGE gel electrophoresis, and then the protein was purified by nickel-sepharose (NI-NTA) chromatography column.

The results show that the purified protein has a molecular weight of 31 kDa and the amount of purified protein is 1 mg.

Based on previous studies as well as the results of this study, the resulting protein can be considered as a suitable candidate for the cholera vaccine.