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Abstract Background Long noncoding RNAs (lncRNAs) have been recognized as viable prognostic and therapeutic indicators for numerous human malignancies. Nonetheless, the operational roles and fundamental mechanisms of important lncRNAs that impact esophageal squamous cell carcinoma (ESCC) remain predominantly obscure. Recently, lncRNAs have been identified to exert regulatory influence on epithelial-mesenchymal transition (EMT) via intricate interplay with EMT-associated transcription factors (TFs) and signaling pathways. The current experimental study aimed to elucidate the expression of four lncRNAs in ESCC patients and explain their potential involvement in the EMT process and the pathogenesis of ESCC. Methods In the study, the expression levels of lncRNAs (AGAP2-AS1, DLEU2, HMBOX1_1 (AC108449.2), and UGDH-AS1) and mRNAs (TWIST1, MMP13, and CD44S) between fifty ESCC and adjacent normal tissue samples were measured using quantitative real-time PCR. Results The upregulation of CD44S (36%), TWIST1 (52%), DLEU2 (58%), AGAP2-AS1 (62%), and MMP13 (74%), were indicated in ESCC samples, while the downregulation of UGDH-AS1 and HMBOX1_1 were found in 62% and 64% of patients, respectively. The expression levels of lncRNAs and EMT-related markers were found to be significantly correlated in several patient clinicopathological traits (P

Background Preterm birth before 37 th wk of gestation is called premature birth. Corticotropin-releasing hormone (CRH) and CRH-binding protein (BP) act on various maternal and fetal tissues during pregnancy, such as the myometrium, which regulates the transition from the dormant phase of the uterus to the active phase. Studies have shown that mir-200c and mir-181a interact with CRH and CRH-BP. Objective The present study aimed to investigate the expression of mir-200c, mir-181a, CRH, and CRH-BP in women with a history of preterm birth. Materials and Methods In this case-control study, the gene expression level of mir-200c, mir-181a, CRH, and CRH-BP in placental tissue samples obtained from 48 women with a history of preterm labor was assessed in the Mojibian hospital of Yazd, Iran, from January to March 2023. Differences between mir-200c, mir-181a CRH, and CRH-BP gene expressions among cases and controls were assessed. Results The outcomes indicated that the expression of CRH increased with going on to the regular parturition time (p < 0.001). While outcomes indicated, CRH-BP decreased with going on to the regular parturition time (p < 0.001). In addition, the results showed that the expression of mir-181a increased and mir-200c decreased with approaching the normal delivery time (p < 0.001). Conclusion In conclusion, the expressions of mir-200c, mir-181a, CRH, and CRH-BP were dissimilar in different weeks of gestation. It could be proposed to use mir-200c, mir-181a, CRH, and CRH-BP as biomarkers to weigh the exact delivery time, which could minimize the side effects of preterm labor for the mother and fetus.

Iron (Fe) deficiency-induced chlorosis can be a serious problem in grapevine cultivation. Measuring genotypic tolerance to this disorder is of vital importance in rootstock selection. In this research, different sources of Fe (FeSO4·7H2O and FeEDTA) were used and their effects were evaluated on the response of a tolerant (‘Thompson Seedless’) and a sensitive (‘Flame Seedless’) grape cultivar. Also, their cross-grafted combinations [‘Flame Seedless’/’Thompson Seedless’ (FS/TS) and ‘Thompson Seedless’/‘Flame Seedless’ (TS/FS)] were evaluated for tolerance to Fe deficiency. Bicarbonate (30 mM) was used for inducing iron deficiency in the plants. Despite the negative effects of bicarbonate, the ‘Thompson Seedless’ maintained a significant chlorophyll index, compared to the ‘Flame Seedless’ cultivar. Gas exchange measurements showed that the bicarbonate had more adverse effects on photosynthesis and water-use-efficiency (WUE) in the ‘Flame Seedless’ cultivar. Despite the bicarbonate toxicity, the photosynthetic system of the FS/TS graft combination showed no significant change in FV/FO, whereas adverse effects occurred significantly in ‘Thompson Seedless’, ‘Flame Seedless’, and their TS/FS graft combination. Bicarbonate decreased the performance index in ‘Flame Seedless’. In conclusion, scions of the susceptible cultivar performed well when grafted on tolerant rootstocks. Such grafting combinations can help plants to cope with the negative effects of bicarbonate toxicity on grapevine.

T helper 9 (Th9) cells, a subset of CD4+ T helper cells, have emerged as a valuable target for immune cell therapy due to their potential to induce immunomodulation and tolerance. The Th9 cells mainly produce interleukin (IL)-9 and are known for their defensive effects against helminth infections, allergic and autoimmune responses, and tumor suppression. This paper explores the mechanisms involved in the generation and differentiation of Th9 cells, including the cytokines responsible for their polarization and stabilization, the transcription factors necessary for their differentiation, as well as the role of Th9 cells in inflammatory and autoimmune diseases, allergic reactions, and cancer immunotherapies. Recent research has shown that the differentiation of Th9 cells is coregulated by the transcription factors transforming growth factor β (TGF-β), IL-4, and PU.1, which are also known to secrete IL-10 and IL-21. Multiple cell types, such as T and B cells, mast cells, and airway epithelial cells, are influenced by IL-9 due to its pleiotropic effects.

Abstract Background The MALAT1 lncRNA acts as an oncogene in Prostate cancer (PC); thus, it can be severe as a cancer biomarker. Methods Using bioinformatics datasets including (HTSeq-Counts, GDC, and TCGA) 5501 gene expression profiling specimens were gathered. Then, expression profiles and sample survival of lncRNA were investigated using COX regression analyses, ROC curve analysis. The Database for Annotation, Visualization, and Integrated Discovery was used to conduct GO and KEGG studies on the lncRNA-related PCGs. After MALAT1 Knockout via CRISPR/Cas9 technique, the MALAT1 expression was assessed in DU-145 cells. The deletion of the target fragment was examined by polymerase chain reaction (PCR). Also, the expression of apoptosis genes was investigated by qRT-PCR. The viability and cell proliferation were measured using the MTT assay. Cell migration capability was determined using the cell scratch assay. The results of qRT-PCR were assessed by the ΔΔCt method, and finally, statistical analysis was performed in SPSS software. Results A maximum of 451 lncRNAs were discovered to reflect different expressions between PC and non-carcinoma tissue samples, with 307 being upregulated and 144 being down-regulated. Thirty-six lncRNAs related to OS were carefully selected, which were then subjected to stepwise multivariate Cox regression analysis, with 2 lncRNAs (MALAT1, HOXB-AS3). MALAT1 is highly expressed in PC cells. MALAT1 Knockout in DU-145 cells increases apoptosis and prevents proliferation and migration, and DU-145 transfected cells were unable to migrate based on the scratch recovery test. Overall, data suggest that MALAT1 overexpression in PC helps metastasis and tumorigenesis. Also, MALAT1 knockout can be considered a therapeutic and diagnostic target in PC. Conclusion Targeting MALAT1 by CRISPR/Cas9 technique inhibit the cell proliferation and migration, and in addition induce apoptosis. Thus, MALAT1 can act as a tumor biomarker and therapeutic target.

Abstract Aim Increased IgE levels have made house dust mite allergens one of the most frequent causes of allergies worldwide. Treatment reduces the IgE antibodies and types two cytokines, namely interleukin‐4 (IL‐4) and IL‐13. Although existing treatments significantly reduce IgE or IL‐4/IL‐13, they are very costly. This study aimed to construct a recombinant protein derived from rDer p1 peptides in the form of an immunotherapy approach and to measure the response of IgE and IgG antibodies. Methods The proteins were isolated, purified, and evaluated using the SDS‐PAGE and Bradford test and confirmed by using Western blot. To evaluate immunotherapy efficiency, 24 BALB/C mice were sensitized intraperitoneally with house dust mites (HDM) adsorbed to Aluminum hydroxide (Alum) and randomly divided into four groups of six: control sensitized, HDM extract, rDer p1, and DpTTDp vaccine. To immunization, four groups of random mice were each treated with phosphate‐buffered saline, 100 μg of rDer p1 protein, DpTTDp, or HDM extract, every 3 days. Direct ELISA determined HDM‐specific IgG and IgE subclasses. Data were analyzed in SPSS and Graph pad prism software. Values of p

CRISPR-Cas9 is the most important tool in genome engineering in recent years. The efficiency of this instrument on active and non-active genes is variable. Programmed cell death protein 1(PD-1) is a surface acceptor on T cells, B cells, and dendritic cells. This protein has an important role in the production of inducing tolerance in lymphocytes. Nowadays, this characteristic is used in cell therapy and immunotherapy of cancer. In the present study, the peripheral blood mononuclear cells and HEK293 cells were selected as expression and non-expression cells of the PD-1 gene. Six pairs of sgRNA were designed for the PD-1 gene. The transfected cells were sorted by the FACS machine. A common pair of primers were used for amplification of cute regions. Px458 was used as an expressional vector for the transfection of PBMCs and HEK293. Transfection was done using lipofectamine and electroporation methods. In PBMCs, 2 guides, sgRNA (3+1) and sgRNA (3+5) were able to disrupt the PD-1 gene. In contrast, in HEK293, none of the 6 guides were able to disrupt it. According to the results obtained, the PD-1 gene cutting in HEK293 cells was failed. However, it was successful in PBMCs. Therefore, it can be told that the heterochromatin region or other genome remodeling mechanisms such as epigenetic remodeling inhibit the PD-1 gene cutting by CRISPR-Cas9 in HEK293 cells.

Photosynthesis is one of the most important physiological processes that is negatively affected by salinity stress in plants. Nitric oxide is known as an influential signal molecule in plant responses to environmental stresses. The tomato plant (Lycopersicon esculentum L.) contains a wide range of vitamins and nutrients that are economically and nutritionally important. In the present study, the effects of different levels of salinity stress including 0, 50, 100 and 150 mM NaCl and sodium nitroprusside (SNP) at 0, 100 and 200 μM levels on different photosynthetic parameters were investigated by JIP test. The results of this study showed that salinity stress reduced the efficiency of the oxygen-evolving complex in photosystem II. Salinity reduces the efficiency of light reactions of the photosystem II (φPo/(1-φPo)) and the efficiency index of active reaction centers (γRC/(1-γRC)). Furthermore, the efficiency of biochemical reactions of electron transport (ψo/(1- ψo)) and the efficiency index of reduction of the end electron acceptor in photosystem I (δRo/(1-δRo)) decreased with salinity stress. The results also revealed that salinity stress reduced the overall performance of the photosynthetic apparatus from the beginning of photosystem II to the end of photosystem I (PITotal). Salinity stress reduced photosynthetic pigments, including chlorophyll a, chlorophyll b and total chlorophyll, and a significant increase in total carotenoids, total polyphenolic content and total flavonoid were observed under salinity stress. The use of SNP as a nitric oxide donor mitigated the effects of salinity stress in all the mentioned parameters. The results of this study showed that in the photosynthetic apparatus of the tomato plants, the electron transport chain is the most sensitive point to salinity stress and 100 μM SNP is the most effective concentration to alleviate the effects of salinity stress.

Coma is a state of prolonged unconsciousness. Some coma cases result from inherited disorders such as fatty-acid β-oxidation disorder, acute intermittent porphyria (due to mutations in genes CPT I, CPTII and ACADM), urea cycle defects (due to mutation in OTC gene), organic acidurias, mitochondrial diseases and familial hemiplegic migraine (due to mutations in CACNA1A, ATP1A2 and SCN1A). The evaluation of familial cases of coma or sporadic coma can be performed using next generation sequencing (NGS), a high-throughput sequencing technique that can sequence an entire genome in a single reaction. This technique has been widely applied in the genetic diagnosis of diseases. In this review, we describe some genes associated with coma or recurrent coma and discuss the role of NGS in detection of these genes.

Allergen-specific immunotherapy (AIT) involves administering allergen extracts. It is used to desensitize allergic patients. Herbal allergen extracts that are optimum in efficacy and fewest in side effects are still challenging to produce. To overcome these limitations, oral immunotherapy, epicutaneous immunotherapy, intralymphatic immunotherapy, and artificial recombinant allergen preparations have been evaluated. Recombinant allergens have become more popular with the development of molecular diagnostics and therapeutics. Besides food and drug allergens, pollen, fungal spores, and other allergens have been studied. Based on related clinical studies, this comprehensive overview will present the latest perspectives on AIT methods and available allergenic products, as well as discuss the challenges and opportunities for treating allergic disorders.

Objective(s): Checkpoint blocking is considered a revolutionary method in cancer treatment. This method eliminates cancer cells by maintaining the sensitivity of immune cells. Today, cell therapy through checkpoint blocking is known as the most efficient method of cancer treatment. The programmed cell death protein-1(PD-1), as an immune check protein, has a vital role in weakening the immune responses by reducing the number of stimulated T cells. In normal situations, a decline in the immune responses can cause induced tolerance and prevent autoimmune diseases.Materials and Methods: In this study, to reduce the induction of tolerance due to PDL-1 binding to PD-1, the PD-1 gene was destroyed in PBMCs by the means of CRISPR-Cas9 and dual-transfection of two plasmids containing the Cas 9 gene and two different sgRNAs specific to two region of PD-1 gene in order to produce a deletion mutation. Six different sgRNA were designed and cloned in PX-458 plasmid vector, and PBMCs were transfected using lipofectamine 2000 and electroporation. Indels were evaluated by gel electrophoresis and Sanger sequencing. Results: We showed the PD-1 gene in PBMCs was knocked out successfully by CRISPR-Cas9 and dual-transfection of two sgRNAs. The minimum interval between the two sgRNAs was 448 nucleotides. Conclusion: The results of this research demonstrated that the use of dual-transfection of CRISPR-Cas9 sgRNA is a suitable method to knock out the PD-1 gene and prevention of inducing tolerance in PBMCs.

Background: Pro-inflammatory cytokines are associated with systemic inflammatory responses. Objective: To investigate the levels of pro-inflammatory cytokines (IL-1b, IL-6, and TNF-a) in patients with non-alcoholic fatty liver (NAFL) and non-alcoholic steatohepatitis (NASH) compared to healthy individuals. Methods: This case-control study was conducted on 30 patients with NAFL, 30 patients with NASH, and 30 healthy volunteers. The plasma level of IL-1b, IL-6, and TNF-a were determined by ELISA, and biochemical parameters were measured using colorimetric methods. Results: IL-1b and IL-6 levels were significantly higher in patients with NASH compared with NAFL and control group. However, TNF-a levels had no significant variations in NAFL and NASH patients compared to the control group (p=0.903 and p=0.960, respectively). Conclusion: Results showed that the levels of ALT activity and pro-inflammatory cytokines were higher in patients with NASH compared to control and NAFL subjects; Therefore, steatosis and inflammation develop as a result of excessive pro-inflammatory factors in NASH.

Salinity is one of the most important abiotic stresses that affect plant growth and development. In order to study the effects of salinity stress on oat (Avena sativa L.), salinity treatments including 0 (control), 50, 100, and 150 mM sodium chloride and nitric oxide in form of sodium nitroprusside in three concentrations (0, 25 and 50 µM) were applied. The changes of chlorophyll fluorescence were analyzed using the JIP-test. Results of this experiment showed that concentrations of 50 and 100mM of Nacl did not have a remarkable effects on chloropyll a fluorescence. Significant increase in chlorophyll a fluorescence was observed in highest salinity (150 mM of Nacl). Furthermore, results of this study also revealed that the application of nitric oxide van alleviates the harmful effects of salinity on photosynthetic electron transport chain where application of sodium nitroprusside in concentration of 50 mM, led to decrease in chlorophyll a fluorescence intensity compared to non-treated plant under salinity stress.

Objective(s): Studies have confirmed that microgravity, as a mechanical factor, influences both differentiation and function of mesenchymal stem cells. Here we investigated the effects of simulated microgravity on neural differentiation of human adipose-derived stem cells (ADSCs). Materials and Methods:We have used a fast rotating clinostat (clinorotation) to simulate microgravity condition. Real-time PCR and flow cytometry analysis were used to evaluate the regulation of neurotrophins, their receptors, and neural markers by simulated microgravity and their impact on neural differentiation of cells. Results: Our data revealed that simulation microgravity up-regulated the expression of MAP-2, BDNF, TrkB, NT-3, and TrkC both before and after neural differentiation. Also, the neural cells derived from ADSCs in microgravity condition expressed more MAP-2, GFAP, and synaptophysin protein in comparison to the 1G control. Conclusion: We showed that simulated microgravity can enhance the differentiation of mesenchymal stem cells into neurons. Our findings provide a new strategy for differentiation of ADSCs to neural-like cells and probably other cell lineages. Meanwhile, microgravity simulation had no adverse effects on the viability of the cells and could be used as a new environment to successfully manipulate cells.

MicroRNAs (miRNAs), have been documented to perform a key role in the pathogenesis of multiple sclerosis (MS), a chronic inflammatory and autoimmune disease. Recent studies have shown that single nucleotide polymorphism in the sequence of the miRNA may change their production and expression which can lead to miRNA dysfunction and pathogenicity. Some studies have reported the relationship between miRNA polymorphism and the increased risk of autoimmune disease. This study was conducted to investigate the association between mir155 rs767649, mir196a2 rs11614913 and mir23a rs3745453 polymorphism and the risk of multiple sclerosis in the Iranian MS patients in Isfahan. A population of 80 patients and the same number control were selected. After DNA extraction, genotyping was performed through tetra amplification refractory mutation system-PCR method (T ARMS PCR). The frequencies of TT, TC and CC genotypes of mir23a were 46, 35 and 20% in MS patients and 42, 14 and 24 in healthy subjects respectively. These results showed that individuals carrying the genotypes of rs3745453 TC had a 2.3-fold increased risk of MS (OR=2.3, p=0.048). There was no significant difference between genotypes and allele frequency of mir155 and mir196a2 in patients and healthy controls (p>0.05). Our findings specified that CT heterozygosity in mir23a gene significantly related with risk of MS. Unlike mir155 and mir196a2, mir23a rs3745453 may have contributed to the etiology of MS in Isfahan patients. However, extensive studies are required to gain more reliable and authentic results.