Discovery medicine最新文献

Background: Although evidence exists on the potential involvement of circular RNAs (circRNAs) in the pathogenesis of several viral infections, the expression levels, and the exact role that hsa_circ_0006459 and hsa_circ_0015962 could play during the Dengue virus (DENV) infection are still unclear. These two circRNAs were identified as possible biomarkers for diagnosis and prognosis of DENV disease in peripheral blood mononuclear cells (PBMC) of Dengue-positive patients. This study aimed to evaluate the expression levels of hsa_circ_0006459 and hsa_circ_0015962 in DENV-infected patients and compare them with healthy donors (HD) to provide new insights into the biological significance of these two circRNAs' expression.

Methods: We examined the presence and expression levels of hsa_circ_0006459 and hsa_circ_0015962 in PBMC of DENV-patients throughout a period of 28 days after the DENV diagnosis. HD was used as a control group.

Results: Our results show different expression levels and patterns between hsa_circ_0006459 and hsa_circ_0015962, both in DENV patients and HD.

Conclusion: Possible change in the hsa_circ_0006459 expression during DENV infection was observed, mainly at the time of diagnosis, but without a consistent pattern among patients during follow-up. Further studies are needed to clarify their expression levels and function both in Dengue-positive patients and HD.

Background: Hereditary nonpolyposis colorectal cancer (HNPCC) is an autosomal dominant disease caused by germline mutations of human DNA mismatch repair (MMR) genes. A significant proportion of HNPCC cases are attributed to large genomic rearrangements of MMR genes, but this finding has been less frequently reported in Chinese populations.

Methods: Array-based multiplex ligation-dependent probe amplification (array-MLPA) was employed in this study to detect genomic rearrangements of 82 probands of Chinese HNPCC families.

Results: According to the results, 18 probands harbored germline genomic deletions of MutL homolog 1 (MLH1) and MutS homolog 2 (MSH2) genes, accounting for approximately 22% (18/82) of the total subjects. Meanwhile, MSH6 gene deletion occurred only in about 2.4% of the probands (2/82). The deletions of MLH1, MSH2 and MSH6 genes were confirmed by classic MLPA analysis, with a concordance rate of 95.5% (21/22).

Conclusion: Array-MLPA is a highly efficient and precise method for clinical screening and diagnosis of HNPCC. By using this method, we found that the HNPCC families carry deletions of MLH1 and MSH2 genes, which are the major germline genomic aberrations in the studied probands. Nevertheless, the deletion of the MSH6 gene is considered a rare occurrence in Chinese HNPCC families, according to our researche. Despite that, it is of clinical significance to screen and diagnose the HNPCC at the early phase by detecting the germline genomic large aberrations in MSH2/MLH1 genes.

Background: Notoginsenoside R1 (NGR1) is a bioactive compound of Panax notoginseng (Burk.) F.H. Chen (PNS), which possesses desirable properties in bone fracture healing and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs). Whether NGR1 can promote osteogenic differentiation of human dental pulp stem cells (DPSCs) is still unknown. This study aimed to assess the biocompatibility of NGR1 and its impact on DPSCs.

Methods: DPSCs were obtained from human wisdom teeth. Flow cytometry and multilineage differentiation were applied to determine stem cell properties. Then, the cells were treated with NGR1 for 1, 2 and 3 days, and its efficacy was detected by means of a 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Alizarin red staining (ARS), alkaline phosphatase (ALP) activity, quantitative calcium node analysis, western blot and reverse-transcription-quantitative polymerase chain reaction (RT-qPCR) were executed to detect osteogenic differentiation-related proteins and genes. Western blot was also performed to assess the activation levels of the p38 mitogen-activated protein kinase (p-38 MAPK), c-Jun N-terminal kinase mitogen-activated protein kinase (JNK MAPK), and extracellular signal-regulated protein kinase mitogen-activated protein kinase (ERK MAPK) pathways in DPSCs following treatment with NGR1.

Results: DPSCs were positive for CD105 and CD166, while negative for CD34 and CD45. NGR1 at concentrations of 10 and 100 μg/mL did not exhibit cytotoxicity (p > 0.05), the group of cells receiving 200 μg/mL and 500 μg/mL NGR1 exhibited proliferation inhibition on the second day as well as on the third day (p < 0.05). Compared to the control group (no treatment), the cells treated with 100 μg/mL NGR1 exhibited significantly higher ALP expression and calcium deposition. The 100 μg/mL NGR1 group also showed higher expression of Osterix (OSX), Runt-related transcription factor 2 (RUNX2), Collagen Type I (COL-1), and Osteocalcin (OCN) at both protein and gene levels. Western blot analysis revealed that NGR1 activated the MAPK pathway by upregulating p38 and ERK, but not JNK, in DPSCs. When the p38 and ERK signaling pathways were inhibited by SB203580 and U0126, the gene expression levels of OSX, RUNX2, COL-1, and OCN were significantly decreased (p < 0.05), but such alterations were not observed with the inhibition of the JNK pathway.

Conclusion: At the concentration of 100 μg/mL, NGR1 enhances DPSC osteogenic differentiation by regulating the MAPK pathways.

Background: One of the pharmacological effects of celastrol (Cel) is the amelioration of acute liver injury. In this study, we explored the mechanism of Cel underlying the alleviation of liver injury induced by traumatic hemorrhagic shock (THS).

Methods: The THS model was developed from Sprague-Dawley rats through transverse fractures, blood loss and fluid infusion. Then, the THS rats were intraperitoneally injected with 0.5, 1, and 1.5 mg/kg Cel. The rats were injected in the tail vein with lentivirus-mediated small interfering RNA (siRNA) negative control (siNC), siRNA targeting heat shock transcription factor 1 (siHSF1), and siRNA targeting toll-like receptor 9 (siTLR9) 72 hours before the establishment of THS model. Hematoxylin-eosin (HE) staining was performed to highlight the pathological alterations in the rat liver tissue. Enzyme-linked immunosorbent Assay (ELISA) was utilized to determine the expression levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and total bilirubin (TB). The expression levels of B-cell lymphoma 2 (Bcl2) and B-cell lymphoma 2 associated X protein (Bax) were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). The expression levels of reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH), and superoxide dismutase (SOD) were determined to assess the extent of oxidative stress. Western blotting was used to evaluate the expression levels of heat shock transcription factor 1 (HSF1), toll-like receptor 9 (TLR9) and myeloid differentiation factor 88 (MyD88).

Results: Cel was shown to therapeutically alleviate liver injury, decrease ALT and AST levels, and simultaneously downregulate inflammation factors levels (TNF-α, IL-1β), alleviated apoptosis, and decreased oxidative stress in the THS model in a concentration-dependent manner. Moreover, Cel increased the expression of HSF1 and decreased the expression of TLR9 and MyD88 in the THS model. And silencing HSF1 increased TLR9 and MyD88 expression. Further, the silencing of HSF1 resulted in liver injury, inflammation and apoptosis, which could be reversed by TLR9 silencing.

Conclusions: This study demonstrates that Cel attenuates THS-induced liver injury by positively regulating HSF1 so as to inhibit the expression of TLR9.

Background: Periodontitis is an immunoinflammatory disease. Ferroptosis is a type of inflammation-associated cell death. The article aims to investigate the expression, role, and mechanism of the ferroptosis-related gene acyl-CoA synthetase long-chain family member 6 (ACSL6) in periodontitis Methods: Ferroptosis-related genes were identified using the Gene Expression Omnibus dataset and the Kyoto Encyclopedia of Genes and Genomes pathway. ACSL6 expression was validated using quantitative reverse-transcription polymerase chain reaction in patients with periodontitis. Human periodontal ligament fibroblasts (hPDLFs) were isolated and characterized. Following treatment, related experiments were performed to evaluate iron levels, reactive oxygen species (ROS) production, cell viability, ACSL6 expression, and ferroptosis-related proteins in hPDLFs.

Results: In this study, 185 genes were upregulated, and 102 were downregulated in the periodontitis group (p < 0.05). ACSL6, a ferroptosis-related gene, exhibited high expression levels in periodontitis tissues (p < 0.05). Porphyromonas gingivalis lipopolysaccharide (P. gingivalis-LPS) upregulated ACSL6 (p < 0.05), downregulated ferroptosis-related genes (glutathione peroxidase 4 (p < 0.001) and cystine/glutamate transporter (Solute Carrier Family 7 Member 11) (p < 0.01)) and phosphor (p)-AMP-activated protein kinase (AMPK) (p < 0.05), reduced cell viability (p < 0.001), and elevated iron (p < 0.001) and ROS levels (p < 0.001) in hPDLFs. ACSL6 silencing could counteract the effects of P. gingivalis-LPS (p < 0.01). Furthermore, AMPK inhibitors lessen the effect of ACSL6 silencing (p < 0.01).

Conclusions: The ferroptosis-related gene ACSL6 was highly expressed in periodontitis tissues, and ACSL6 silencing enhanced viability and inhibited ferroptosis in P. gingivalis-LPS-mediated hPDLFs by upregulating the AMPK pathway.

Background: Calycosin is thought to have anti-cancer and anti-inflammatory characteristics; however, more research is needed to determine how it impacts retinal pigment epithelium (RPE) cells. This study aims to explore the effects of calycosin on RPE cells under hypoxia.

Methods: Experimental hypoxia was induced by treating RPE cells with cobalt chloride for 2, 4, and 6 h. To investigate the effect of calycosin on RPE cells under hypoxia, RPE cells were treated with calycosin and cobalt chloride (CoCl₂). Cells were assessed for viability (Cell Counting Kit-8 assay) and apoptosis (flow cytometry). Inflammatory cytokines (enzyme-linked immunosorbent assay) and genes or proteins related to apoptosis and the hypoxia-inducible factor-1α (HIF-1α)/nuclear factor-κB (NF-κB) axis (quantitative real-time polymerase chain reaction and western blot) were measured.

Results: Under hypoxic conditions, RPE cells showed reduced viability but increased levels of inflammation and apoptosis. The NF-κB pathway was activated, and HIF-1α, apoptosis/NF-κB pathway-related proteins (cleaved caspase-3, cleaved poly (ADP-ribose) polymerase (PARP); phosphorylated-p65 (p-p65), p-p65/p65), and inflammatory cytokines (interleukin-6 (IL-6) and interleukin-8 (IL-8)) were upregulated (p < 0.001). Calycosin weakened the effects of hypoxia on RPE cells (p < 0.05).

Conclusion: Calycosin inhibits the HIF-1α/NF-κB axis and protects RPE cells from hypoxia-induced inflammation and apoptosis.

Over the recent years, immunomodulators have opened a new avenue in cancer treatment by virtue of their ability to boost the immune system for neoplastic cell elimination. Improving treatment outcomes by leveraging the interaction of these agents with traditional cancer treatments is the main emphasis of this review. Checkpoint inhibitors, chemokine receptors, and pattern recognition receptors are the immunological targets of their interactive mechanisms. Immunomodulators are generally categorized as inhibitors of checkpoint, cytokines, agonists, or adjuvants. Despite their high efficacy and specificity, modern-day antibody-based therapies face several key limitations such as immunogenicity, insufficient tissue penetration, and restricted oral bioavailability. To address these shortcomings, researchers are crafting small molecules with the potential for oral administration and improved pharmacokinetic properties. These agents can augment antibody therapies for synergistic effects to enhance therapeutic efficacy for different types of cancers. This review explores the synergy between immunomodulators and traditional cancer treatments (chemotherapy, radiation, and targeted therapies) as well as newer strategies like adoptive cell therapies (chimeric antigen receptor therapies such as chimeric antigen receptor-T (CAR-T) cell therapy and chimeric antigen receptor-natural killer (CAR-NK)). These combinations improve treatment effectiveness in a number of ways: radiotherapy increases tumor antigen presentation and T-cell infiltration, chemotherapy-induced immunogenic cell death boosts immune responses and targeted therapies lessen immunosuppression in the tumor microenvironment. Despite the potential appeal as adjuvants, immunomodulators also pose challenges in maximizing their efficacy and minimizing adverse effects. In this paper, clinical trials proving the effectiveness of these combined techniques are reviewed, and innovative approaches including next-generation checkpoint inhibitors and delivery systems based on nanoparticles are also highlighted. Overall, this review evaluates the existing impact of immunomodulatory adjuvants and their prospective trends in cancer care. Further development of immunomodulators will pave the way for more accessible and effective therapies, marking a significant step towards personalized oncological interventions.

Corneal disorders, encompassing injuries, infections, and degenerative diseases, are major contributors to visual impairment globally. Conventional procedures, including corneal transplantation and pharmacological treatments, encounter constraints such as donor shortages, rejection risks, and diminished effectiveness in extreme instances. Mesenchymal stem cells (MSCs) have emerged as viable therapeutic alternatives owing to their regeneration potential, immunomodulatory characteristics, and capacity to differentiate into corneal cell types. This study examines the therapeutic potential of MSCs in addressing various corneal illnesses through the analysis of preclinical studies, clinical trials, and current breakthroughs. MSCs facilitate corneal wound healing, diminish scarring, and reinstate transparency via processes including paracrine signaling, extracellular matrix remodeling, and anti-inflammatory actions. Although early-phase clinical trials indicate the safety and feasibility of MSC-based therapeutics, obstacles persist in optimizing delivery techniques, assuring cell viability, and creating uniform protocols. Additional research is necessary to address these issues and validate MSCs as a feasible clinical alternative. This review aims to summarize the therapeutic applications, challenges, and future prospects of mesenchymal stem cells in corneal treatments, emphasizing their importance as emerging alternatives to traditional therapies.

Background: Tumor-initiating cells (TICs) play a pivotal role in the unfavorable outcomes of laryngeal tumor proliferation, recurrence, and resistance to chemoradiotherapy. This study aims to explore the expression of CD271 (p75 neurotrophin receptor (p75^NTR) in human laryngocarcinoma Hep2 cells and unravel its potential biological functions as a marker of laryngeal TICs.

Materials and methods: Immunomagnetic cell sorting was utilized to separate subsets of Hep-2 cells based on high and low expression levels of CD271. Various aspects such as proliferation activity, colony formation ability, cell cycle distribution, and the expression of cancer-related proteins in each subpopulation were evaluated using immunofluorescence, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, soft agar gel assay, flow cytometry, and western blot assay. Furthermore, the tumor-forming potential of the subsets displaying high and low CD271 expression was examined through an in vivo experiment involving nude mice. The proteins associated with the phosphorylated signal transducer and activator of transcription 3 (p-STAT3)/Octamer-binding transcription factor 4 (OCT4) pathway were detected via western blot assay.

Results: The expression of CD133 was the highest in the CD271 high-expression group, and the expression of CD133 was the lowest in the CD271 low-expression group. Hep2 cells with high CD271 expression exhibited enhanced proliferation capacity, in contrast to those with low CD271 expression which showed reduced proliferation (p < 0.05). The CD271 high-expression group of Hep2 cells demonstrated superior clonogenic ability, a higher proportion in the S and G2/M phases of the cell cycle, and an increased sphere-forming capacity. Moreover, Hep2 cells with high CD271 expression displayed enhanced tumor formation capability in nude mice (p < 0.001). Western blot analysis indicated significantly elevated levels of specific proteins such as OCT4, Nanog Homeobox (NANOG) and p-STAT3/STAT3 in the CD271 high-expression group were significantly higher than those in the control group (p < 0.01), and the protein levels of low-expression group were significantly lower than those in the control group (p < 0.01).

Conclusions: CD271 serves as a marker for TICs in Hep-2 cells, presenting a novel target for further investigation.

Background: Glucosamine-6-phosphate N-acetyltransferase 1 (GNPNAT1) is an enzyme involved in the hexosamine biosynthetic pathway, which is critical for glycosylation processes. In the context of non-small cell lung cancer (NSCLC), GNPNAT1 plays a significant role in modulating immune responses. The purpose of this study is to investigate the role of GNPNAT1 in regulating the efficacy of radiotherapy and resistance to natural killer (NK) cell-mediated cytotoxicity in patients with NSCLC.

Methods: To assess GNPNAT1's impact on radiotherapy efficacy, 122 lung cancer patients were categorized into radiosensitive and radioresistant groups. GNPNAT1 expression levels in cancerous tissues from both groups were measured using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blotting. This analysis was extended to various lung cancer cell lines (BEAS-2B, A549, LTEP-2, SPCA1, and H157) using the same molecular techniques. To investigate GNPNAT1's functional role in radioresistance, radioresistant A549 cells (A549R26-1) were established, and GNPNAT1 expression was genetically manipulated. Experimental groups included control, si-NC, si-GNPNAT1, Oe-NC, and Oe-GNPNAT1. Post-treatment, GNPNAT1 levels were measured via qRT-PCR and Western blotting. Cells were exposed to varying doses of radiation, and subsequent assessments included cell proliferation (Cell Counting Kit-8 (CCK-8) assay), radiosensitivity (plate cloning assays), and apoptosis rates (flow cytometry). Isolated and purified primary NK cells were co-cultured with lung cancer cells from each experimental group. The cytotoxicity of NK cells against lung cancer cells was assessed through lactate dehydrogenase (LDH) release and colony formation assays.

Results: Compared to the radiosensitive group, the radioresistant group exhibited significantly elevated GNPNAT1 expression levels (p < 0.05). The radioresistant cell line A549R26-1 demonstrated higher proliferation ability and lower apoptosis levels compared to its parental cell line, A549P. Subsequently, down-regulation of GNPNAT1 expression in A549R26-1 cells resulted in reduced proliferation, increased apoptosis, and weakened resistance to NK cell cytotoxicity. Conversely, up-regulation of GNPNAT1 expression in A549R26-1 cells following co-culture with NK cells led to increased proliferation and survival rates, and enhanced resistance to NK cell cytotoxicity. Notably, GNPNAT1 knockdown effectively attenuated the radioresistance of A549R26-1 cells.

Conclusion: Down-regulation of GNPNAT1 expression reduces the immune resistance of non-small cell lung cancer to radiotherapy and enhances susceptibility to NK cell cytotoxicity.