Cell Journal最新文献

Veno-occlusive disease with immunodeficiency (VODI) syndrome is a rare genetic disorder characterized by immune system irregularities and a significant mortality rate, despite its infrequency. SP110, situated on chromosome 2q37.1, plays a pivotal role in VODI syndrome, and its association with tuberculosis has been extensively studied. The identification of SP110 mutations holds promise for accelerating the diagnosis and treatment of VODI syndrome, by providing a comprehensive panel for diagnosis and potentially leading to targeted therapies. In this case study, we examined a three-year-old girl born to a consanguineous union who was suspected of having an immunodeficiency disorder. Whole-exome sequencing (WES) and clinical assessments were conducted to screen for and confirm potentially pathogenic mutations. The detected mutation was further analyzed using bioinformatics tools to forecast its impact on protein structure. WES analysis revealed a novel deletion-insertion mutation, c.1181-1182delAGinsT, within SP110. Protein analysis indicated substantial structural modifications in the SP110 protein. This study identified a novel deletion-insertion mutation as a potential contributor to VODI syndrome by affecting the functionality of the SP110 protein. By including various mutations associated with the SP110 gene, this study aimed to expedite diagnosis by creating a comprehensive panel for VODI syndrome.

Objective: Polycystic ovary syndrome (PCOS) is one of the most important causes of infertility, irregular menstrual cycles, and anovulation in women. The current study aimed to investigate the therapeutic effects of Althaea officinalis L. (A. officinale) extract on PCOS in rats.

Materials and methods: In this experimental study, 70 rats in 7 groups (n=10/group) were studied for three weeks as follows; healthy control (HC), patient (PCOS), metformin (PCOS+MET), A. officinale treatment (PCOS+250 and 500 mg/kg A. officinale) and synergistic (PCOS+MET+250 and 500 mg/kg A. officinale) groups. Luteinizing hormone (LH), folliclestimulating hormone (FSH), progesterone (P) and testosterone (T) levels as well as inflammatory cytokines were measured. Total antioxidant capacity and lipid peroxidation levels were analyzed in ovarian tissue. The expression of GLUT-4, AKT, PI3K, PTEN genes and Ki-67 was assessed by real-time polymerase chain reaction (PCR) and immunohistochemistry, respectively.

Results: A. officinale alone and especially in combination with MET moderated inflammatory and antioxidant parameters compared to the PCOS and MET groups. A. officinale in synergistic groups increased the apoptosis of granulosa cells by activating the PI3K/AKT pathway, resulting in a rise in the number of Ki-67 positive cells (P<0.05). Furthermore, following A. officinale treatment the LH/FSH rate decreased and FSH and P increased (P<0.05). Also, A. officinale extract could effectively normalize estrus cycle duration close to the normal group.

Conclusion: The extract of A. officinale, in combination with metformin, can enhance the hypothalamic-pituitaryovary (HPO) axis with synergistic anti-inflammatory and antioxidant effects. Additionally, the extract showed apoptotic effect on cystic granulosa cells.

Objective: CD22, as a surface protein of B cells, is used in the diagnosis and target-specific immunotherapy of B-cell malignancies. SpyTag and SpyCatcher, on the other hand, are two covalently coupled proteins capable of developing a bi- or multi-specific modular protein. The aim of this study was to develop FITC-conjugated SpyCatcher-SpyTagged anti-CD22 Nanobody (FITC-SpyC-SpyT-CD22Nb) to recognize CD22 on the surface of malignant B cells.

Materials and methods: In this experimental study, the SpyTag-CD22Nb construct was subcloned into a pET22 vector and expressed in E. coli BL21 (DE3). After purification using His-tag affinity chromatography, the size of the eluted protein was confirmed on a Western blot. In addition, the SpyCatcher protein, subcloned into pET28, was expressed in E. coli BL21 (DE3), purified by His-tag affinity chromatography and subjected to FITC labeling. FITC-SpyCatcher and SpyTag-CD22Nb were coupled in a 1:1 molar ratio. The specific binding of the produced FITC-SpyC-SpyT-CD22Nb was tested using CD22+ Raji and CD22- K562 cell lines and was evaluated by flow cytometry.

Results: SpyTag-CD22Nb and SpyCatcher were successfully expressed in E. coli BL21 (DE3). The 1:1 molar ratio of SpyTag-CD22Nb and FITC-SpyCatcher successfully formed FITC-SpyC-SpyT-CD22Nb at room temperature. The flow cytometry results showed that FITC-SpyC-SpyT-CD22Nb specifically binds to the CD22+ Raji cells, while there is no binding to the CD22- K562 control cells.

Conclusion: The novel FITC-SpyC-SpyT-CD22Nb produced in the present study is capable of detecting the surficial expression of CD22. According to our findings, FITC-SpyC-SpyT-CD22Nb is applicable for specific targeting of CD22 in a therapeutic manner, i.e., chimeric antigen receptor (CAR)-T cell therapy and antibody drug conjugates (ADCs).

Objective: The spliced form of X-box binding protein 1 (XBP1s) is a key transcription factor in the unfolded protein response (UPR), an adaptive mechanism for cell survival. Many studies demonstrated the induced expression of XBP1s in various cancers, including hepatocellular carcinoma (HCC). Such upregulated expression is linked to an enhancement of cell proliferation, migration, and improvement of the survival rate. In this study, we aimed to assess the therapeutic potential of targeting XBP1s, by specific decoy oligodeoxynucleotide (ODN) and evaluated the cancerous phenotypes in Huh-7 cells.

Materials and methods: In this experimental study, we transfected Huh-7 cells with XBP1s decoy oligonucleotide (ODN). Subsequently, we assess some cellular features, including viability, migration capacity, proliferation potential, and apoptosis. Therefore, various techniques included wound healing test, BrdU, and annexin/PI assays. Additionally, the colony formation capacity was evaluated. The mRNA expression levels of BAX, BCL-2, c-MYC, CCND1, MMP-9, CDH1, and CD133 were quantified by the reverse transcription-quantitative polymerase chain reaction (RT-qPCR).

Results: Transfection of Huh-7 cells by XBP1s decoy ODN led to significant down-regulation of c-Myc, CCND1, MMP-9, BCL-2 and CD133 and up-regulation of CDH1 and BAX transcriptional expressions in comparison with the vehicle group. Our results also demonstrated that transfection of XBP1s-decoy reduced HCC cell viability, proliferation, migration capacity as well as colonization ability in comparison with the vehicle group.

Conclusion: These findings proposed the potential application of XBP1s-decoy ODN to reduce cancerous phenotypes such as cell proliferation, cell migration and apoptosis induction in the Huh-7 cell line. More experiments on other cell lines and primary cells could validate our results.

Objective: Breast cancer is a prevalent and heterogeneous disease, with human epidermal growth factor receptor-2 (HER2) overexpression occurring in over 20% of cases. Poncirin, a biologically active flavonone derived from the immature dried fruits of Poncirus trifoliata, is a 7-O-neohesperidoside of isosakuranetin with a well-documented history in traditional Chinese medicine for its health-promoting properties. While the previous research hinted at its potential as an anticancer agent, its specific effects on HER2 overexpressing breast cancer cells remain largely unexplored. The aim of this study is to investigate the specific effects of Poncirin, on HER2 overexpressing breast cancer cells.

Materials and methods: In experimental study, we assessed cell proliferation using the CCK-8 assay and explored cell migration and invasion with transwell assays. Additionally, we evaluated colony formation ability and examined apoptosis through the acridine orange/ethidium bromide (AO/EB) and Annexin V-fluorescein isothiocyanate (FITC)/ propidium iodide (PI) staining methods. The study also delved into the molecular mechanisms involved by scrutinizing the phosphatidylinositol 3-kinase/serine-threonine protein kinase (PI3K/AKT) signaling pathway via Western blotting. Furthermore, the researchers conducted in vivo experiments using mouse models to corroborate the findings in a living organism.

Results: Poncirin demonstrated a remarkable ability to selectively inhibit proliferation and metastasis of HER2 overexpressing breast cancer cells. Mechanistically, the compound seemed to exert its effects by modulating the PI3K/AKT signaling pathway, implying its central role in the observed anticancer effects. These findings were further substantiated by in vivo experiments, which consistently showed a reduction in tumor growth when poncirin was administered.

Conclusion: This study underscores potential of poncirin as a potent agent for restraining the growth and metastasis of HER2 overexpressing breast cancer cells. The evidence suggests that poncirin efficacy may be attributed to its modulation possibly through PI3K/AKT pathway.

Given the critical role of human papillomavirus (HPV) in the cause of cervical cancer and other malignancies, there is a need for innovative approaches to preventing this infection. It has been shown that immunoinformatics is an important strategy in computational vaccinology. It is used to design new multi-epitope vaccines against different types of HPV and subsequent cervical cancer. This paper reviews the scope of the entire computational pipeline of HPV vaccine design, starting from data analysis at the genomic and proteomic levels and continuing to epitope predictions of the innate and adaptive immune systems. The search strategy was based on investigating original articles published in "Google Scholar" and "PubMed" from 2015 to 2023-2024. The terms "Immunoinformatics", "Bioinformatics", "Human papillomavirus (HPV)", "Vaccine design", "In silico vaccine design", "Multi-epitope vaccine design", "Vaccinology" and "HPV vaccine" were used to for this purpose. We discussed various essential tools involved in the computational design of the vaccine process, e.g., sequence analysis, epitope prediction, conservancy analysis, tertiary structure modeling, refinement, molecular docking, molecular dynamics (MD) simulation, and in silico cloning. This review article describes immunoinformatics methods that facilitate the design of a multi-epitope vaccine against HPV. However, this pipeline can also be used to design novel chimeric vaccines for other pathogens.

Objective: Head and neck squamous cell carcinoma (HNSCC) with a high mortality rate is among the most common types of cancer in the world. Human epidermal growth factor receptor 2 (HER2) is expressed higher than normal level in the most HNSCC tumors, making them resistant to chemotherapy and radiotherapy. Therefore, HER2 has been introduced as a suitable target for anticancer drugs. The aim of this study is to examine the efficacy of a treatment protocol involving targeted delivery of idarubicin encapsulated in trastuzumab-decorated liposomes to HNSCC cells.

Materials and methods: In the current experimental study, efficacies of idarubicin, prepared liposomal idarubicin, and constructed immunoliposomal idarubicin (trastuzumab-decorated) were investigated in killing HN5 cells, a HER2- overexpressing HNSCC-originating cell line. Liposomal content of idarubicin and trastuzumab were qualified by UVVisible spectroscopy and preparations were characterized for shape and size by atomic force microscopy (AFM) and dynamic light scattering (DLS). To clarify role of the missing parts of the available crystal structure (PDB ID: 1n8z) within trastuzumab-HER2 interactions, we used a 40 ns molecular dynamic simulation approach.

Results: Based on the obtained results, liposomal idarubicin showed higher toxicity of the encapsulated drug on HN5 cells compared to the traditional free drug formulations. The immunoliposomal form of idarubicin was more effective than the liposomal formulation, in killing of HN5 cells. In addition, simulation of interactions between trastuzumab and HER2 revealed that the missing parts (in the crystal structure) of HER2 have critical interaction with trastuzumab, through salt-bridges and hydrogen bonds.

Conclusion: It seems that the prepared immunoliposomes could attach more efficiently to HER2 overexpressing cells, which consequently leads to increasing cellular uptake of idarubicin through a receptor-mediated endocytosis mechanism. Moreover, simulation of the interaction between HER2 and trastuzumab suggested considerable possibilities for increasing trastuzumab affinity to HER2.

Damaged articular cartilage has limited self-healing potential and often leads to osteoarthritis (OA), pain, and dysfunction of the affected joint. Autologous and allogenic transplants cannot fully meet the needs of clinical treatment. Two dimensional (2D) and three-dimensional (3D) cell cultures can help to study growth modeling and physiological characteristics of the human body. Among the problems that 2D and single-layer cultures have the lack of proper and accurate tissue modeling and the lack of tissue complications similar is to the original tissue. With organoid models, cellular and tissue structural studies and functional and physiological studies of tissues have been revolutionized and more accurate. Organoids are useful for studying repair and drug efficacy. Physiological and pathological investigations by combining in vitro and in vivo methods have become more effective today. The purpose of this study is to investigate the factors involved in the formation of cartilage organoids so that we can introduce the best method of organoid production for the healing of cartilage damage by using cell types and organoid model.

Objective: Cardiovascular diseases (CVDs) are the leading cause of death worldwide, with atherosclerosis serving as a primary factor in their development. Platelets, leukocytes, and their interactions play a crucial role in initiating and amplifying atherosclerosis. This study aims to evaluate the levels of platelet-monocyte aggregates (PMA) and specific integrins involved in leukocyte recruitment, including macrophage-1 antigen (Mac-1) and lymphocyte functionassociated antigen-1 (Lfa-1), in patients with acute coronary syndrome (ACS).

Materials and methods: In this case-control study, thirty-two subjects with ACS and 30 healthy individuals participated. It aimed to evaluate PMA expression and the median fluorescence intensity (MFI) of Mac-1 and Lfa-1 using flow cytometry. Dot plots and Pearson correlation coefficient were employed to examine the relationship between PMA, Mac-1, and Lfa-1. Multilevel model analysis was used to explore the effects and relationships of various parameters, including Mac-1 and Lfa-1, on PMA. Finally, receiver operating characteristic (ROC) curves were utilized to assess the diagnostic accuracy of PMA, Mac-1, and Lfa-1 markers.

Results: It was observed that patients had higher PMA levels compared to the control group (58.99 ± 16.27 vs. 29.99 ± 4.19 in controls, P<0.001), which correlated with PLT (ρ=0.512, P=0.035). Additionally, CD18 and CD11b expression on monocytes were significantly elevated in patients (P<0.001) and were positively associated with PMA (β=19.09, P<0.001; β=6.90, P=0.022), but no significant relationship between CD11a and PMA was observed (β=5.06, P=0.315). PMA and Mac-1 were identified as better markers for differentiating patients from healthy individuals. (respectively, AUC=0.94, Sensitivity= 0.84, specificity=0.98; AUC=0.84, Sensitivity= 0.93, specificity=0.70).

Conclusion: The study results indicated an increase in both Mac-1 and PMA levels in patients with ACS. Additionally, the significant association observed between Mac-1 and PMA in the patient group suggests a potential relationship between these markers and ACS.

Objective: The purpose of this study was to investigate the effects of 8 weeks of high-intensity interval training (HIIT) and vitamin D3 supplementation on Chemokine (C-C motif) Ligand 5 (CCL-5) and C-C motif chemokine receptor 5 (CCR5) in the white adipose tissue (WAT) of male rats with type 2 diabetes (T2DM).

Materials and methods: The experimental study involved 40 male Wistar rats divided into 5 groups (n=8). These groups were healthy control (HC), diabetic control (DC), diabetic+HIIT (DHIIT), diabetic+vitamin D3 (DD3), and diabetic+HIIT+vitamin D3 (DHIITD3). The rats completed 8 weeks of HIIT, consisting of 12 sessions lasting 1 minute each at an intensity of 90-95% of their maximum running speed. Additionally, the rats were administered a weekly dose of 10,000 IU/kg of vitamin D3 for 8 weeks.

Results: The levels of CCL-5 (P<0.001) and CCR5 (P=0.003) were found to be higher in the DC group as compared to the HC group. However, when HIIT training and vitamin D3 were administered together, there was a decrease in CCL-5 (P=0.001) and CCR5 (P<0.001) in the DHIITD3 group (P=0.001). Similarly, vitamin D3 alone reduced CCR5 levels in the DD3 group (P< 0.001). Also, the decrease of CCR5 in the DD3 group was higher than in the DHIIT group (P=0.022), and the DHIITD3 group was higher than in the DHIIT group (P<0.001), but there was no difference between the DD3 and DHIITD3 groups (P≥0.05).

Conclusion: The results indicate that combining HIIT training with vitamin D3 has a greater effect on reducing the expression of CCL-5 and CCR5 in the white adipose tissue of rats with type 2 diabetes induced by streptozotocin (STZ) and a high-fat diet (HFD), compared to the effects of each one alone. It is recommended that the study be conducted by measuring the variables involved in the mechanisms and the changes in CCL-5 and CCR5.