Cellular and molecular biology最新文献

Oral squamous cell carcinoma (OSCC) is the most frequently diagnosed oral malignancy and poses a great threat to public health. According to bioinformatics analysis, long noncoding RNA PCBP1-AS1 is downregulated in OSCC. In this work, the functions and mechanism of PCBP1-AS1 in OSCC were further investigated. PCBP1-AS1 expression in OSCC cells was measured by quantitative polymerase chain reaction. Cell viability and proliferation were detected using CCK-8 assays and colony-forming assays. TUNEL assays as well as flow cytometry analyses were carried out to detect OSCC cell apoptosis. Binding relationship between PCBP1-AS1 and miR-34c-5p or that between miR-34c-5p and ZFP36 in OSCC cells was identified using RNA immunoprecipitation assays, RNA pulldown assays, and luciferase reporter assays. Experimental results revealed that PCBP1-AS1 was downregulated in OSCC cells. PCBP1-AS1 overexpression hampered cell proliferation and enhanced cell apoptosis in OSCC. PCBP1-AS1 interacted with miR-34c-5p in OSCC and negatively regulated miR-34c-5p. ZFP36 3'untranslated region was targeted by miR-34c-5p. PCBP1-AS1 positively regulated ZFP36 expression. ZFP36 silencing abrogated the suppressive impact of PCBP1-AS1 on OSCC cell growth. In summary, PCBP1-AS1 suppresses cell growth in OSCC by upregulating ZFP36 through interaction with miR-34c-5p.

Caffeine is believed to exert its therapeutic effects by acting as a nonselective, competitive antagonist of adenosine receptors. Chlorpromazine, a phenothiazine, is a classic psychotropic mediator extensively used in the clinical administration of psychotic disorders. This study aimed to validate the procedures used for performing Functional Observational Battery (FOB) tests, to demonstrate the proficiency and interobserver reliability during the FOB tests and also to assess effect on neurobehavioral parameters using positive controls in rats. The rats were administered with Caffeine in Milli-Q water as oral gavage at the dose of 20 mg/kg and Chlorpromazine HCl in 0.9% Saline as intraperitoneal route at the dose of 20 mg/kg. No inter-personnel variability was observed in home cage, handling, open field and sensory reactivity observations recorded in Proficiency test. In conclusion, the known effects of positive controls; caffeine and chlorpromazine HCl on neurobehavioral/Functional Observational Battery parameters including autonomic, neuromuscular and sensory reactivity tests were detected in the current study. FOB test procedures for neurobehavioral, grip strength and motor activity are adequate for the detection of neurotoxic effects of positive controls. No major inter-personnel variability was observed between study personnel in neurobehavioural observations.

This study aimed to explore the influence and mechanism of low-intensity pulsed ultrasound (LIPUS) combined with Rhodiola bone penetration on the formation of spinal fusion bone. Sixty clean-grade New Zealand white rabbits were selected for randomization and divided into combined group and Rhodiola group, with 30 rabbits in each group to construct a rabbit lumbar intervertebral fusion model, using Rhodiola intervention and Rhodiola combined with LIPUS intervention protocol, respectively. The axial strength, axial stiffness, maximum compressive load, vascular endothelial growth factor (VEGF), cycloxygenase-2 (COX-2), prostaglandin E2 (PGE2) and transforming growth factor-β (TGF-β) were compared after HE staining, immunohistochemistry and biomechanical detection. Spine fusion rate was 100.00%; the combined bone graft tissue had implanted bone cell degeneration, cell necrosis and cell hyperplasia, chondrocytes differentiated into trabecular bone and some hematopoietic cells, severe cell necrosis and fiber cell proliferation and late bone formation in the Rhodiola group, VEGF, COX-2, PGE2, TGF-β, axial strength, axial stiffness, and maximum compression load in the combined group significantly increased (P<0.05). Spinal fusion using LIPUS combined with Rhodiola can enhance biomechanical properties and promote the role of PGE2, COX-2, VEGF, TGF-β expression and bone formation, and this protocol is worthy of clinical application.

We aimed to explore the potential along with mechanism of lncRNA growth arrest-specific 5 (GAS5) in modulating glucose metabolism and ferroptosis of endothelial progenitor cells (EPCs) in coronary heart disease (CHD). CCK-8, flow cytometry, EdU, colony formation, scratch test as well as transwell assays were implemented to assess cell biological behaviors. Glucose uptake testing, lactic acid production assay, and detection of extracellular acidification rate (EACR) together with oxygen consumption rate (OCR) were used to assess glucose metabolism. Iron, GSH and MDA detection were used to measure ferroptosis. Besides, a series of mechanical experiments were implemented to clarify the modulatory relationship between GAS5 and nuclear factor erythroid 2-related factor 2 (NRF2) as well as sine oculis homeobox 1 (SIX1). We found that GAS5 was down-regulated in CHD patients relative to healthy controls. GAS5 depletion repressed EPCs proliferation, migration along with invasion while elevated cell apoptosis. GAS5 promoted the reprogramming of glucose metabolism and inhibited ferroptosis in EPCs. GAS5 affected glycometabolic reprogramming and ferroptosis resistance through regulating SIX1 and NRF2. On the one hand, GAS5 promoted NRF2 mRNA stability through IGF2BP2. On the other hand, GAS5 regulated the miR-495-3p/SIX1 axis in EPCs. To sum up, GAS5 promotes glucose metabolism reprogramming and resistance to ferroptosis of EPCs through the miR-495-3p/SIX1 and IGF2BP2/NRF2 dual-regulatory pathways in CHD.

Triple-negative breast cancer TNBC with higher immunogenicity and tumor-infiltrating lymphocyte (TIL) enrichment can benefit from immunotherapy relative to other breast cancer subtypes. Our work was designed to identify the TIL-related hub genes in TNBC and construct a prognostic signature for TNBC. TNBC gene expression files were obtained from the TCGA database. CIBERSORT algorithm and random forest risk model were used for immune infiltration group division. The TIL-related differentially expressed genes (DEGs) were then selected and subject to GO, KEGG analyses and GSEA. Next, Lasso cox regression analyses were adopted for constructing a prognostic risk model, followed by evaluation using time-dependent ROC curves. The copy number variation between the two risk groups was also analyzed, and major genomic mutation types were identified. Additionally, the nomogram was constructed with calibration curve for clinical prognosis analysis. Our results showed that totally 113 TNBC samples were allocated into the high or low-immune risk groups. We identified 243 DEGs between groups, namely TIL-related DEGs, with 128 upregulated and 115 downregulated genes. Among the TIL-related DEGs, 6 hub genes (SLITRK3, PCDHGB3, NELL2, SRRM4, ASIC2 and B4GALNT2) were screened out and constructed a prognostic risk signature, which had good performance for long-term prognosis prediction. Analysis of genomic mutation showed that the TP53, PIK3CA, TTH, etc. showed high mutation frequency in the two prognostic risk groups. Moreover, the higher risk score of the prognostic risk model predicted poor overall survival in TNBC patients, and nomogram and calibration curve confirmed the potent prediction ability of this model. To sum up, six TIL-related biomarkers (SLITRK3, PCDHGB3, NELL2, SRRM4, ASIC2 and B4GALNT2) were identified and used for the construction of the prognostic risk model, which might provide novel insight for the clinical decisions.

Dactylobiotus parthenogeneticus is one of the widespread species of tardigrade all over the world. Tardigrades of this species were collected from the Greater Zab River in Erbil City-Iraq by filtering water of the river through a plankton net with a mesh of 45 µm pore. The samples were mounted on a slide with a cover slip and examined under the microscope to determine morphological characteristics and measurements. Based on these characters the species identified to be D. parthenogeneticus. To support this diagnosis, DNA barcoding techniques were applied to do molecular analysis and sequencing on the cytochrome oxidase subunit I (COI) gene. The sequence was subjected to the GenBank database of NCBI and recorded with the accession number PP140905. The result of the sequencing and molecular analysis of the cytochrome oxidase subunit I (COI) gene confirmed to be the same species diagnosed by relying upon morphological characters. This study represents one of the pioneer researches and documents on tardigrades and found D. parthenogeneticus for the first time in the Greater Zab River in Kurdistan, North of Iraq. Tardigrades play a magnificent role in different trophic levels and can be utilized as an indicator of ecosystem health.

Finding novel, efficient antimicrobial drugs is crucial in this age of pressing global health challenges. The medicinal qualities of the leaves of the argel plant (Solanum argel, or S. argel) have been recognized in traditional medicine for quite some time. The medicinal potential of these leaves may be due to the presence of bioactive substances such as alkaloids, flavonoids, and phenolic acids. S. argel leaf antibacterial, phytochemical, and gas chromatography-mass spectrometry (GC-MS) characteristics are the focus of this investigation. To conduct the study, bioactive compounds would be extracted from the leaves and tested against a panel of bacterial pathogens. Then, the compounds would be identified using GC-MS analysis. Mean inhibition zones of 15.30±1.0 mm, 14.67±0.42 mm, 15.0±0.01 mm, and 15.56±0.22 mm for the bacteria E. coli, Staph. aureus, and Sal. typhimurium, respectively, were seen in the antibacterial results at a concentration of 3 µg/disc. Secondary metabolites such as alkaloids, flavonoids, phenolic substances, and tannins were identified using phytochemical investigation. Antimicrobial, antioxidant, and anti-inflammatory are just a few of the many bioactivities associated with these phytochemicals. Argel plant leaves contain bioactive chemicals that show they could be a source of new pharmaceuticals. Argel leaves were analyzed using GC-MS and 37 different chemicals were found. The most abundant compounds were 4H-Pyran-4-one and 2,3-dihydro-3.5-hydroxy, followed by 3-Pentanol, 2,2,4,4-tetramethyl, and 2,2-Dimethyl-3-[3-methyl-5-(phenylthio)-, with areas of 11.80%, 10.6%, and 9.47%, respectively. The analysis was performed within a time range of 5.070 to 34.464 minutes. According to the research, Argel leaf has powerful antioxidant and antibacterial capabilities, making it an excellent substance for medical and food preservation applications.

Following the outbreak of COVID-19, several immunotherapy methods were used to modulate the immune responses of patients. In this study, we aimed to evaluate the immune response to COVID-19 in patients receiving convalescent plasma. In this regard, this randomized controlled trial included 30 patients who were divided into two groups according to receiving convalescent plasma or normal control plasma. Samples from both groups were collected on days 0, 1, 3, 5 and 7 after plasma infusion. We measured the expression level of TLR7/8, IRF3/7, CTLA-4, PD-1 and T cell transcription factors by Real-time PCR in the mentioned groups. Thirteen cytokines were also evaluated using flow cytometry method. Results showed that compared to the normal control plasma group, the expression levels of TLR7, 8, IRF3, 7 and PD-1 and CTLA-4, on days 3, 5 and 7 after convalescent plasma infusion, were significantly decreased. On the other hand,  Gene expression results showed that the expression levels of Tbet, RORγ3 and Foxp3 on days 3, 5 and 7 after convalescent plasma infusion were significantly increased compared to the normal control plasma group. After convalescent plasma infusion, the viral load was significantly decreased compared to the normal control plasma group. Convalescent plasma infusion also reduced the plasma cytokines levels, including IL-6, IL-10, and IL-4, and enhanced the level of IL-2, IFN- γ and perforin comparing the normal control plasma group. According to the results, the convalescent plasma infusion led to a decrease in the expression of innate immunity receptors and an increase in the expression of transcription factors of adaptive immunity. Therefore, it may be concluded that convalescent plasma infusion can modulate the immune response. To achieve a reliable consequence, further studies are required.

Human Papilloma Virus (HPV) is an oncogenic virus and is the most common cause of cervical cancer. HPV has been shown to induce senescence. Cellular senescence is involved in cancer progression and tumorigenesis. Identification and isolation of cells of tumor origin before tumorigeneses is an important step in cancer prevention and treatment. This study aimed to investigate the early cervical atypical senescent cytological preneoplastic change in non-menopausal women. Cervical smears of 121 patients were randomly selected and included in the study which cytopathologically diagnosed as atypical squamous cells of undetermined significance (AS-CUS) in correlation to HPV status, parakeratosis (PK), p16 immunostaining, enlarged Squamous cells nuclei (ES) and inflammatory cells infiltration (ICI). Results revealed that out of the total 121 patients, 32 cases (26%) were positive for high-risk HPV (HR-HPV), 26 cases (22%) were positive for low-risk HPV (LR-HPV) and 63 (52%) were negative for HPV. HPV infections were significantly associated with age groups (p<0.026), PK (p = 0.043), p16 (p = 0.001), ES (p = 0.002) and ICI (p = 0.049). The positive immunostaining expression of p16 was only noticed in two HR-HPV patients. ES cells were found in 9.5% of HPV-negative cases, 27% of LR-HPV cases and 40.5% of HR-HPV cases. High PK cell positivity was seen only in HR-HPV. High ICI scores were seen in 40.6% of HR-HPV patients, 26.9 % of LR-HPV and 17.4 % of negative HPV patients. It was concluded that high PK positivity, high ICI score, positive p16 immunostaining and ES were correlated with HR-HPV in non-menopausal women. These findings could provide potential diagnostic clues for HPV-harboring senescent cells as a strategy for reducing HPV risk of cervical cancer development and identifying the cell of tumor origin, which could be beneficial for improving the utility of senolytic agents and immunotherapy in clinical practice.

This experiment aimed to evaluate the impact of several dentine etching and conditioning agents on growth factors (GFs) liberation from dentine slices. Eighteen dentine slices were obtained from nine premolars divided in to six groups, the slices immersed in one mL test solutions for 5 min; Group 1: white Mineral trioxide aggregate (MTA), Group 2: Phosphate buffered saline (PBS), Group 3: 37% phosphoric acid, Group 4: 17% Ethylenediaminetetraacetic Acid (EDTA), Group 5: 10% Maleic acid (MAc), and Group 6: 0.7% Fumaric acid. The solutions were removed and stored directly at for further detection and quantification of transforming GF beta 1 (TGF-b1), bone morphogenetic protein 2 (BMP2) and vascular endothelial growth factor (VEGF) by enzyme-linked immunosorbent assay (ELISA). One-way ANOVA was used to compare the mean release and standard deviation between groups (α = 0.05). Tukey's post hoc applied for multiple comparisons. After five min conditioning of dentine slices, white MTA released the highest level of TGF-b1, BMP2 and VEGF among all groups, followed by 0.7% Fumaric acid with no significant difference between them, but compared to 37% phosphoric acid and PBS groups significant difference observed, which they released the least amount of GFs amongst all groups. Based on the results of this research the detectable release of TGF-b1, BMP2 and VEGF by 0.7% fumaric acid was comparable with white MTA from dentin slices.