Cellular and Molecular Biology

miR-378a-5p represses Barrett’s esophagus cells proliferation, migration and invasion through targeting TSPAN8

Xudong Xie — 2024-02-28

Barrett’s esophagus (BE) belongs to a pathological phenomenon occurring in the esophagus, this paper intended to unveil the underlying function of miR-378a-5p and its target TSPAN8 in BE progression. GEO analysis was conducted to determine differentially expressed genes in BE samples. Non-dysplastic metaplasia BE samples, high-grade dysplastic BE samples and controls were collected from subjects. CP-A and CP-B cells were exposed to bile acids (BA) to mimic gastroesophageal reflux in BE cells. RT-qPCR as well as western blot were applied for verifying expressions of miR-378a-5p, TSPAN8, CDX2 and SOX9. CCK-8, wound scratch together with Transwell assays were exploited for ascertaining cell proliferation, migration as well as invasion. The targeted relationship of miR-378a-5p and TSPAN8 could be verified by correlation analysis, dual-luciferase reporter experiment, and rescue experiments. Through analyzing GSE26886 dataset, we screened the most abundantly expressed gene TSPAN8 in BE samples. miR-378a-5p was reduced whereas TSPAN8 was elevated in CP-A as well as CP-B cells after triggering with BA. Knocking down TSPAN8 could counteract BA-triggered enhancement in BE cell proliferation, migration along with invasion. miR-378a-5p could suppress BE cell proliferation, and migration along with invasion via targeting TSPAN8. In BE, miR-378a-5p targeted TSPAN8 to inhibit BE cell proliferation, and migration along invasion. miR-378a-5p deletion or elevation of TSPAN8 may be key point in regulating CDX2 and SOX9 levels, thereby promoting BE formation.

Changes in biochemical and sperm parameters of rats drinking energy drinks

Sarwar N. Jafar — 2024-02-28

Tiger energy drink is used in this research to see the effect on serum lipid profile, liver enzymes, and kidney function, also observing changes in sperm parameters. Sixteen male rats were divided into four groups, each consisting of four rats. The control group was given water and rat feed, while groups 1, 2, and 3 were given water with Tiger energy drink (25%, 50%, and 75%, respectively) for eight weeks. The results showed that the effect of energy drinks on lipid profile decreased cholesterol, TG, and LDL, while there was a slight increase in HDL. Treatment with wild Tiger energy drink generally caused insignificant increases in GOT and GPT in both G2 and G3 groups as compared to the control. However, in the G1 group, GPT and GOT significantly increased. Accordingly, ALP concentration was significantly increased in all experimental groups compared to the control group. Sperm quality declined as the concentration of energy drinks consumed increased. The potential adverse effects of energy drinks, at their specific dosages, have been determined to include hepatic and renal impairment, as well as alterations in serum lipids. Additional research is required to investigate the specific mechanism by which energy drinks impact spermatogenesis, either through the modulation of endocrine hormones or other metabolic pathways.

METTL3-mediated m6A modification enhances ZDHHC16 expression in nonsmall- cell lung cancer patients, attenuating ferroptosis by suppressing CREB ubiquitination

Zeyu Liu — 2024-02-28

At present, the early diagnosis and treatment of non-small cell lung cancer (NSCLC) is still an urgent problem to be solved worldwide, including in China. The present work investigated the possible protective effect of ZDHHC16 in cell proliferation and metastasis of NSCLC and explored its possible mechanisms. ZDHHC16 expression level in patients with Non-Small-Cell Lung Cancer was up-regulation. ZDHHC16 gene is stabilized by m6A methylation. ZDHHC16 gene reduced ferroptosis of NSCLC by the rehabilitation of the mitochondrial structure. ZDHHC16 promoted CREB expression through the inhibition of CREB Ubiquitination. Confocal microscopy showed that ZDHHC16 reduced the CREB expression of NSCLC. ZDHHC16 up-regulation reduced CREB Ubiquitination, and down-regulation of ZDHHC16 promoted CREB Ubiquitination of NSCLC. CREB Agonists reduced the effects of ZDHHC16 on ferroptosis, not affecting the Warburg effect of NSCLC. CREB inhibitor reduced the effects of si-ZDHHC16 on ferroptosis, not affecting the Warburg effect of NSCLC. METTL3-mediated m6A modification increases ZDHHC16 stability. Our study revealed that the m6A-forming enzyme METTL3 upregulates ZDHHC16 expression in NSCLC patients, leading to the reduction of ferroptosis by inhibiting CREB ubiquitination.

Circ-LDLRAD3/miR-655-3p/MAPK1 axis enhances cell migration and invasion in papillary thyroid carcinoma

Bingfei Zhang — 2024-02-28

Papillary thyroid carcinoma (PTC) is a prevalent histological subtype of thyroid cancer, whose occurrence and development may be related to circRNA dysregulation. This research proposed to unravel circ-LDLRAD3-related mechanisms in PTC. First, circ-LDLRAD3, miR-655-3p .and MAPK1 levels in PTC were quantitatively measured. Then, plasmid vectors or oligonucleotides that interfere with circ-LDLRAD3, miR-655-3p, or MAPK1 were transfected into PTC cells, followed by the analysis of proliferation, apoptosis, migration, and invasion. Finally, the targeted binding sites between miR-655-3p and circ-LDLRAD3 or MAPK1 were predicted by starBase and experimentally verified. Statistically, PTC samples expressed high circ-LDLRAD3 and MAPK1 and low miR-655-3p. Knocking down circ-LDLRAD3 or enhancing miR-655-3p hindered PTC cell proliferation, migration, and invasion, and forced apoptosis. circ-LDLRAD3 bound to miR-655-3p to affect MAPK1 expression. Elevating MAPK1 rescued circ-LDLRAD3 knockdown-allowed obstruction of PTC cell growth. In conclusion, circ-LDLRAD3 stimulates PTC development by releasing miR-655-3p-targeted MAPK1.

The role of mir96 in predicting CTC status and prognostic evaluation in gastric cancer patients

Qi Zheng — 2024-02-28

In this study, 20 patients clinically diagnosed with advanced gastric cancer were selected as subjects. Circulating tumor cells (CTCs) in the peripheral blood of gastric cancer patients were detected and counted by collecting peripheral blood samples at Ningbo No. 2 Hospital and using the Cell Rich TM system combined with a negative enrichment strategy. In addition, routine pathological examination and immunohistochemical staining were performed on surgical specimens, including HER2, EBER, E-cadherin, and vimentin indicators. These indicators were correlated and analyzed with CTC counts and routine clinical tests. At the same time, miRNA groups were performed to explore the miRNAs with or without correlation with CTC and to try to construct a predictive model for CTC status. Tumor tissue from patients whose CTC counts did not match the results of the miRNA prediction model was subjected to second-generation gene sequencing to analyze indicators such as tumor heterogeneity and immune microenvironment. The results of the study showed that CTCs were detected in the peripheral blood of 50% (10/20) of gastric cancer patients using the Cell Rich TM system, serum fibrinogen was negatively correlated with CTC counts, whereas TNM staging elements, HER2 receptor and p53 were not correlated with CTC counts. miRNA assays showed that the expression levels of miR218, miR1207, miR96, miR409, miR149, miR148a, miR155, miR370 and miR223 were significantly different between the two groups of CTC≥2 and CTC<2. Among them, miR-96 showed good efficacy as an indicator for predicting CTC status and assisting in determining the prognosis of gastric cancer. In conclusion, the present study demonstrated that the Cell Rich TM system combined with the negative enrichment strategy can effectively detect CTCs in the peripheral blood of gastric cancer patients and that the expression of miR-96 can be used as an effective indicator to predict CTC status and assist in determining the prognosis of gastric cancer. These findings have important implications for the diagnosis and treatment of gastric cancer and provide new clues for the further study of the tumor immune microenvironment and tumor heterogeneity.

Evaluation of coriander seed powder supplementation on body composition, growth performance, hematology, and serum biochemical indices in common carp (Cyprinus carpio) fingerlings

Zena Ahmed Qader — 2024-02-28

This study aimed to investigate the role of coriander seed powder (Coriandrum sativum) on growth indices, feed utilization, body composition, and haemato-biochemical parameters in common carp (Cyprinus carpio) fingerlings over 84 days. One hundred and forty-four common carp (25.24±0.05 g) were assigned into four groups fed with different diets, namely 0 (basal diet), 1%, 2%, and 4% of coriander seed powder (CSP). In the current study, C. carpio fed with dietary CSP revealed significant improvement in weight gain, final weight, specific growth rate, total feed intake, feed conversion efficiency, feed conversion ratio, protein intake, and protein efficiency ratio, in comparison to control fish fed after 84 days (P>0.05). It was also found that fish fed with 1%CSP-supplemented dietary had the best growth performance and feed utilization. The crude protein of fish fed with CSP dietary treatments increased, and significant differences were only found in the fish fed with 1%CSP diet, in comparison to the control group. The CSP supplementation groups showed significant increases in hemoglobin, hematocrit, albumin, total protein, and globulin compared to the control group. Nevertheless, differential white blood cells, mean corpuscular hemoglobin concentration, cholesterols, and triglycerides were significantly reduced in the CSP dietary group in comparison to the control group. It was also found that CSP dietary treatment significantly increased lipase and amylase in comparison to the control group (P>0.05). However, the highest lipase and amylase levels were obtained at 1%CSP and 2%CSP dietary treatment groups, compared to the control basal diet. Based on the results, CSP supplementation could improve the overall health status and growth performance of common carp fingerlings.

Homocysteine modulates CXCL10/CXCR3 axis activity to induce endothelial dysfunction

Yanjie Xu — 2024-02-28

Elevated homocysteine (Hcy) levels have been linked to the development of cardiovascular diseases, notably endothelial dysfunction, a critical precursor to atherosclerosis. In this extensive investigation, we explore the intricate pathways through which Hcy influences endothelial dysfunction, with particular attention to the CXCL10/CXCR3 axis. Employing a dual approach encompassing both in vitro and in vivo models, we scrutinize the repercussions of Hcy exposure on endothelial functionality. Our results reveal that Hcy significantly impairs crucial endothelial processes, including cell migration, proliferation, and tube formation. Concomitantly, Hcy upregulates the expression of adhesion molecules, exacerbating endothelial dysfunction. In a murine hyperhomocysteinemia (HHcy) model, we observed a parallel increase in plasma Hcy levels and adverse vascular effects. Moreover, our study unraveled a pivotal role of the CXCL10/CXCR3 axis in Hcy-induced endothelial dysfunction. Hcy exposure led to the upregulation of CXCL10 and CXCR3, both in vitro and in HHcy mice. Importantly, the blockade of this axis, achieved through specific antibodies or NBI-74330, mitigated the detrimental effects of Hcy on endothelial function. In conclusion, our findings illuminated the central role of the CXCL10/CXCR3 axis in mediating Hcy-induced endothelial dysfunction, providing valuable insights for potential therapeutic strategies in managing HHcy-related cardiovascular diseases.

Diagnostic advancements: Isolating Mycobacterium avium ssp. paratuberculosis and unveiling its molecular identity with nested-PCR

Faranak Nouri — 2024-02-28

Mycobacterium avium subspecies paratuberculosis (MAP) is the causative agent of paratuberculosis, which is currently prevalent in many parts of Iran and produces severe economic loss. It is hence necessary to identify and isolate the animals infected with this bacterium, so this research aimed to isolate MAP from milk and fecal samples of ELISA-positive animals and determine the molecular identity of isolates. After performing ELISA on 3,700 bovine blood samples, 115 samples of milk and feces were taken from ELISA-positive cattle and were cultured on Herald's egg yolk medium with and without mycobactin-J and then the acid-fastness of positive samples was determined using Ziehl–Neelsen staining. The 16S rRNA-PCR test was performed after DNA extraction to determine the molecular identity of isolates. Primers IS6110 and IS901 were employed to ensure that the isolates were not related to members of M. tuberculosis complex and M. avium, respectively. Primer IS900 was also used to determine the molecular identity of MAP isolates. Also, expression levels of MAP-related genes (IS900, ISMAP02, F57, MAP2191, MAP4027) were evaluated via qPCR. Finally, positive samples were confirmed based on the Nested-PCR. Results showed that a total of 9 isolates were obtained from the culture of 90 ELISA-positive samples. The results revealed that all grown samples were positive for acid-fastness. The 16S rRNA-PCR test revealed the 543 bp band, which confirms the presence of Mycobacterium in the samples. The PCR test with Primer IS900 generated the 398 bp fragment in the first step and the 298 bp fragment in the second step, indicating the presence of MAP in samples. Also, relative expression analysis revealed that MAP-related genes were significantly higher in ELIZA-positive samples than in negative ones. Based on the study findings, it can be concluded that MAP-infected animals can be identified by ELISA. In addition, mycobacterium can be isolated by culturing the samples on appropriate media and then its molecular identity can be determined by using nested-PCR.

Determination of volatile aroma composition of some hazelnut varieties grown in Akçakoca conditions

Hulya Unver — 2024-02-28

Türkiye is leading country for hazelnut production and hazelnut orchards are widely established in Black Sea belt. Akçakoca district belongs to Duzce province is accepted as one of the most important production areas. In this research, volatile aroma components in nuts (kernels) of Çakıldak, Kara, Sarı (Mincane), Tombul and Yomra hazelnut varieties grown in Akçakoca district were determined by SPME/GC-MS. The analysis revealed the presence a total of 55 different volatile aroma components. These analyzes identified the presence of 39 descriptive volatile aroma compounds with significant differences between varieties. Çakıldak variety differed from the other varieties in terms of Isopentyl alcohol and Ethyl acetate components. At the same time, some volatile aroma compounds are commonly detected among the all varieties. This study has shown that the climate and soil conditions of Akçakoca cause the emergence of unique taste profiles through the effect of hazelnut varieties on volatile aroma components. The findings emphasize that regional diversity and local ecosystem factors play a critical role in determining flavor in hazelnut kernels. This study also emphasizes that variety selection is a critical factor in ensuring sustainability in hazelnut cultivation and that determining volatile aroma components is an important indicator in this selection.

Transcriptional analysis of oxidative and nitrosative stress on oral opportunistic Candida albicans

Rezhin Ali Mohammed — 2024-02-28

The yeast Candida albicans is one of the most aggressive opportunistic pathogens in immunocompromised patients. The ability of the yeast to withstand stresses and radicals is of great concern. In the present study, four isolates of C. albicans were taken from patients with oral candidiasis and grown on RPMI for 24 hours at 37°C. Then, they were exposed to various concentrations of oxidative (H2O2) and nitrosative (HNO3) stress for two hours, and gene expression rates were measured through RT-PCR. After initial biofilm formation steps and growth validation, RNA extracted from the yeast and gene expression status were evaluated. Upon treatment with H2O2, the gene expression profile for ALS1, MLH1, and EXO1 showed approximately a fold increase in expression. While within HNO3 the yeast gene expression exhibited a dramatic increase in ALS1 up to 217 folds, while others such as MLH1, HWP1, and ERG11 showed a one-fold increase in the expression rate. The findings of this research indicate a considerable expression activity within the biofilm of Candida albicans, increased rate of DNA mismatch repair and break fixation may indicate the ability of the yeast to tolerate high concentrations of free radicals. It paves the way toward understanding the pathogenicity of the yeast and its survival capability inside macrophages. The study also revealed that the biofilm strategy of the yeast is more active within these stresses.

Protective effect of exosomes derived from bone marrow mesenchymal stem cells on hypoxia reperfusion injury of cardiomyocytes

Di Zhao — 2024-02-28

We aimed to investigate the cardiomyocyte-protective effects of bone marrow mesenchymal stem cells (BMSCs)-derived exosomes on ischemia/reperfusion (I/R)-injured rats and to explore the mechanisms. Cardiomyocytes were divided into control group, ischemia/reperfusion group (I/R group), ischemia/reperfusion+exosome group (I/R+Exo group) or ischemia/reperfusion+exosomes transfected with miR-101a-3p inhibitor group (I/R+Exo inhibitor group). MiR-101a-3p levels were lower in I/R and I/R+Exo inhibitor groups than in control and I/R+Exo groups. Apoptosis rate and cleaved caspase 3 expression were higher in I/R and I/R+Exo inhibitor groups. The levels of superoxide dismutase (SOD) in cardiomyocytes of I/R group and I/R+Exo inhibitor group were lower than those of control group and I/R+Exo group, and the levels of malondialdehyde (MDA) and the relative production of oxygen species clusters (ROS) in cardiomyocytes of I/R group and I/R+Exo inhibitor group were higher than those of control group and I/R+Exo group. The levels of interleukin-10 (IL-10), interleukin-6 (IL-6), tumour necrosis factor α (TNF-α), and nuclear factor κB (NF-κB) were higher in the I/R group and the I/R +Exo inhibitor group than in the control group and the I/R+Exo group. Bioinformatics analysis suggested that Pik3c3 is the most promising gene involved in miR-101a-3p-mediated apoptosis in cardiomyocytes, and in vitro experiments confirmed that low expression of miR-101a-3p significantly up-regulated the mRNA and protein expression levels of Pik3c3. BMSCs-derived exosomes have a protective effect on cardiomyocytes from I/R-injured rats, and the mechanism may be related to the inhibition of oxidative stress and inflammatory responses in cardiomyocytes by exosome-delivered miR-101a-3p.

Dynamic changes and clinical value of Sirt6 in acute coronary syndrome (ACS) patients

Gang Tian — 2024-02-28

This study aimed to investigate the role of Sirt6 and inflammatory cytokines in blood samples of patients with ACS. This is a retrospective randomized controlled clinical trial, a total of 30 patients from our hospital are included and divided into following two groups: control group and experimental group, and experimental group consists of 15 patients with ACS and control group consists of 15 patients with non-acute coronary syndrome. Sirt6 protein is detected by western blotting and Sirt6 mRNA is detected by real-time PCR, then inflammatory cytokines such as IL-1β, IL-18, TnI, and CK-MB are measured by ELISA and cytokines NT-proBNP are monitored by immunofluorescence. Our outcomes show that Sirt6 protein and Sirt6 mRNA in experimental group are remarkably lower than those in control group, and IL-1β, IL-18, TnI, CK-MB, and NT-proBNP in the experimental group are remarkably higher than those in control group. We can conclude that Sirt6 can prevent or inhibit the development of ACS and IL-1β, IL-18, TnI, CK-MB, and NT-proBNP can accelerate the development of ACS.

Purification, kinetic characterization of thermostable multicopper oxidase from the oyster mushroom and its versatility for greener agro-pulp bio bleaching in the paper industry

Manmeet Kaur — 2024-02-28

Production of a thermostable laccase from Pleurotus florida was reported for the first time, both in submerged and solid-state fermentation using agro-industrial residues. This enzyme was purified using ammonium sulphate precipitation (60-90%), Sephadex G-100 and DEAE column ion exchange chromatography, respectively. The laccase was purified to 21.49 fold with an apparent molecular weight of 66 kDa and had an optimal pH of 5 with temperature stability at 60°C. Metal ions such as Cu²⁺ (91.26 µmole/mL/min), Mg²⁺ (68.15 µmole/mL/min), and Fe²⁺ (1.73 µmole/mL/min) enhanced the laccase activity, but Fe²⁺ (1.73µmole/mL/min) inhibited the enzyme activity. The purified laccase had K_m and V_max of 16.68 mM and 26.73 µmole/mL/min for guaiacol as a substrate. The isolated enzyme was characterized by FT-IR which revealed bands at 3655.0 cm^-1, 2894.7 cm^-1, and 1151.7 cm^-1 corresponding to primary amines, C-H stretch, and amide –III, respectively. The enzymatic bio bleaching of paddy straw pulp was found to be most effective which resulted in a lowering of kappa number and yellowness by 19.47% & 17.84% whereas an increase in brightness and whiteness by 41.92%. & -19.61%. Thus, this might be stated that the crude laccase from P. florida can be exploited to reduce the toxic waste load for managing environmental pollution and helps in enhancing the yield and quality of the paper.

Synergistic anti-tumor effects of lenalidomide and gefitinib by upregulating ADRB2 and inactivating the mTOR/PI3K/AKT signaling pathway in lung adenocarcinoma

Shuangshuang Zhou — 2024-02-28

Gefitinib is commonly used to be the first-line therapy for advanced non-small cell lung cancer (NSCLC). Therapeutic effect of gefitinib is reduced due to acquired resistance, and combined treatment is recommended. In this research, we planned to explore the impacts of combined treatment of lenalidomide and gefitinib on gefitinib-sensitive or -resistant NSCLC cells. The co-treatment results demonstrated that enhanced antitumor impact on NSCLC cell growth, migration, invasion, cell cycle process and apoptosis. The tumor-bearing mouse models were established using PC9/GR cells. In vivo assays also showed that lenalidomide and gefitinib synergistically inhibited mouse tumor growth along increased the survival of mice. ADRB2 was identified as a lowly expressed gene in PC9/GR cells and LUAD tumor tissues. LUAD patients with high ADRB2 expression were indicated with favorable survival outcomes. Moreover, ADRB2 was upregulated in lenalidomide and/or gefitinib-treated PC9/GR cells. ADRB2 deficiency partially offsets the suppressive impacts of lenalidomide and gefitinib co-treatment on the viability and proliferation of PC9/GR cells. Additionally, lenalidomide and gefitinib cotreatment significantly inactivated the mTOR/PI3K/AKT signaling pathway compared with each treatment alone. Rescue assays were performed to explore whether lenalidomide and gefitinib synergistically inhibited the growth of PC9/GR cells via the PI3K/AKT pathway. PI3K activator SC79 significantly restored reduced cell proliferation, migration and invasion along with elevated cell cycle arrest and apoptosis caused by lenalidomide and gefitinib cotreatment. In conclusion, lenalidomide and gefitinib synergistically suppressed LUAD progression and attenuated gefitinib resistance by upregulating ADRB2 and inactivating the mTOR/PI3K/AKT signaling pathway in lung adenocarcinoma.

Effects of Jerusalem artichoke-enriched diet on water quality, growth performance, feed utilization, proximate body composition, and hematology and biochemical parameters in common carp fingerlings

Samad S. Omar — 2024-02-28

The purpose of this research was to evaluate the impact of Jerusalem artichoke tubers (Helianthus tuberosus L.), a natural prebiotic, on the growth performance, proximate body composition, feed utilization, hematology, and biochemical parameters in common carp (Cyprinus carpio) fingerlings. Four JA-supplemented diets were formulated at 0.0% (control), 0.5%, 1%, and 2%. Fish were reared for ten weeks in cages placed in concrete ponds. Based on the results, an increase in the levels of JA supplementation led to significant improvements in growth and feed parameters (P<0.05), while the proximate body composition exhibited significant differences (P<0.05) between JA-supplemented-fed fish and the control-fed fish. The hematological profile showed that red blood cells, white blood cells, lymphocytes, hematocrit, hemoglobin, and mean corpuscular volume were significantly enhanced by supplementing dietary with JA at varying levels (P>0.05). However, the fish fed with a JA-supplemented diet exhibited significantly lower levels of red cell distribution width, red cell distribution, monocytes, granulocytes, mean corpuscular hemoglobin in fL, and mean corpuscular hemoglobin concentration (P<0.05). Biochemical indices revealed that fish in the experimental groups had significantly higher total protein, globulin, albumin, lipase, high-density lipoprotein, and amylase than the control-fed fish (P<0.05). The creatinine, glucose, triglyceride, cholesterol, urea, alanine transaminase, aspartate aminotransferase, and low-density lipoprotein were significantly decreased in JA dietary treatments than control diet (P<0.05). It was also found that dietary JA supplements promoted growth parameters, proximate body composition, hematology, and serum biochemical in common carp fingerlings.

Pan-cancer analysis reveals a regulatory pattern of anoikis in human cancers

Xingjian Wang — 2024-02-28

Anoikis emerges when a cell finds itself extricated from the appropriate extracellular matrix, leading to an interruption in integrin ligation and thus triggering programmed cellular demise. The cardinal role of Anoikis in the realms of tumor invasion and metastasis is undeniable, although our grasp on its precise influence within the convoluted landscape of cancer biology remains somewhat circumscribed. Notably, both the immune milieu of the tumor and its inherent aggression are correlated with the fluctuating variables of Anoikis. We conducted a thorough evaluation of the genes associated with anoikis and studied the regulatory patterns of these genes as well as the prognostic impact of anoikis in 33 different types of tumors. We provided functional annotations for the regulatory patterns linked to Anoikis. Additionally, we described the associations between immunological factors and genes associated with Anoikis. By applying gene set variation analysis (GSVA), we utilized the inherent abilities of 34 basic genes to calculate the Anoikis index. The Anoikis index is closely related to prognosis, immune microenvironment, immunotherapy, and other aspects. Our functional research revealed a correlation between immune cell infiltration, EMT, and a regulatory gene that is synonymous with adverse survival outcomes. In addition, our observations revealed a direct relationship between the expression of CEACAM5 and CEACAM6,the amplification of epithelial mesenchymal transition (EMT) phenomenon, and a decrease in survival outcomes.The potential therapeutic utility of anoikis-related genes was highlighted by the possible links between TME, clinical samples, genetic mutations, drug resistance, and immunotherapy.

Combination therapy application of Abemaciclib with Doxorubicin in triple negative breast cancer cell line MDA-MB-231

Tugce Nur Eralp — 2024-02-28

Due to lack of clinical biomarkers, Triple Negative Breast Cancer (TNBC) is more likely to have spread to other tissues at time of diagnosis and therapy planning generally involves use of cytotoxic chemotherapy agents, such as Doxorubicin. We aimed to investigate possible advantages of using combination strategy using Doxorubicin alongside Abemaciclib. After determining the IC50 values for Doxorubicin (DOX) and Abemaciclib (ABE); CompuSyn and ComBenefit software were used to reveal the effect resulting from the combination of two drugs. Following the determined effect, cell death was revealed by fluorescence microscopy and a colony forming assay was performed to see the potential of even a single cancer cell with adhesive character to survive over time and form a clone of itself. Detection of changing antioxidant activity following DOX, ABE and DOX+ABE combination therapy in MDAMB231 cells was determined by measuring MDA, SOD and GSH activities. The expression of Cleaved Caspase 3, PARP, Cleaved PARP, Cdk2 and Bax, which changed as a result of DOX, ABE and DOX+ABE application, was shown by Western Blotting.Cyclin-dependent kinase inhibitors appear as promising agents in therapy planning for breast cancer due to their prominent role in cell cycle regulation, where the number of studies interrogating its efficiency in the treatment of cancer such as TNBC is limited. For this reason, in this study, we aimed to determine the impact of the combined use of the CDK4/6 inhibitors ABE and DOX on the cytotoxicity, apoptotic homeostasis, alterations in antioxidative mechanisms, and the molecular pathways that they utilize. Our results showed that when used in combination, Doxorubicin and Abemaciclib showed a synergistic effect on TNBC cell line MDA-MB-231.

Clinical efficacy of early microsurgical clipping of tumor neck in the treatment of cerebral aneurysm rupture and bleeding and its impact on oxidative stress response and prognosis of patients

Zhenglou Chen — 2024-02-29

Cerebral aneurysm can rupture a blood vessel and cause bleeding in the brain. Microsurgical clipping of the tumor neck has been reported to be effective in treating cerebral aneurysm rupture and bleeding. This research attempted to clarify the clinical efficacy of early microsurgical clipping of tumor neck for treating cerebral aneurysm rupture and bleeding, and its impact on the prognosis of patients. One hundred patients with cerebral aneurysm rupture and bleeding patients were treated. They were selected and divided into experimental group (n=25) and control group (n=25) according to surgical time. All patients underwent microsurgical clipping of tumor neck for therapy. The control group chose to undergo surgery 72 hours after the onset of cerebral aneurysm rupture and bleeding, while the experimental group chose to undergo complete surgery within 72 hours after the onset of cerebral aneurysm rupture and bleeding. Primary outcome measures were incidence of complications, cognitive function scores, prognosis, surgical indicators, oxidative stress response and quality of life. Results showed that compared to the control group, the incidence of complications in experimental group exhibited depletion (P<0.05), the prognosis in experimental group exhibited elevation (P<0.05), the hospitalization time in experimental group exhibited depletion (P<0.05), the nomination, abstraction, language, orientation, attention, delayed recall and visual and executive function scores and total scores in experimental group exhibited elevation (P<0.05), serum levels of oxidative stress-related indicators in experimental group exhibited depletion (P<0.05) and the quality of life in experimental group exhibited elevation (P<0.05). In conclusion, early microsurgical clipping of the tumor neck can reduce the risk of complications and cognitive impairment in patients with cerebral aneurysm rupture and bleeding.

Bone mesenchymal stem cell-derived exosomal miR-26a-3p promotes autophagy to attenuate LPS-induced apoptosis and inflammation in pulmonary microvascular endothelial cells

Qianfei Wang — 2024-02-28

Acute lung injury (ALI) is a serious lung disease. The apoptosis and inflammation of pulmonary microvascular endothelial cells (PMVECs) are the primary reasons for ALI. This study aimed to explore the treatment effect and regulatory mechanism of bone mesenchymal stem cell-derived exosomes (BMSC-expos) on ALI. PMVECs were stimulated by Lipopolysaccharide (LPS) to imitate ALI environment. Cell viability was determined by CCK-8 assay. Cell apoptosis was evaluated by TUNEL and flow cytometry. ELISA was utilized for testing the contents of TNF-α, IL-1β, IL-6, and IL-17. Western blot was applied for testing the levels of autophagy-related proteins LC3, p62, and Beclin-1. RNA interaction was determined by luciferase reporter assay. The ALI rat model was established by intratracheal injection of LPS. Evans blue staining was utilized for detecting pulmonary vascular permeability. Our results showed that LPS stimulation notably reduced cell viability, increased cell apoptosis rate, and enhanced the contents of inflammatory factors in PMVECs. However, BMSC-exo treatment significantly abolished the promoting effects of LPS on cell injury. In addition, we discovered that BMSC-exo treatment notably activated autophagy in LPS-induced PMVECs. Furthermore, BMSC-expos upregulated miR-26a-3p expression and downregulated PTEN in PMVECs. MiR-26a-3p was directly bound to PTEN. MiR-26a-3p overexpression reduced cell apoptosis, and inflammation and promoted autophagy by silencing PTEN. Animal experiments proved that miR-26a-3p overexpression effectively improved LPS-induced lung injury in rats. The results proved that BMSC-expos promotes autophagy to attenuate LPS-induced apoptosis and inflammation in pulmonary microvascular endothelial cells via miR-26a-3p/PTEN axis.

Effects of Leukocyte-rich platelet-rich plasma and Leukocyte-poor platelet-rich plasma on cartilage in a rabbit osteoarthritis model

Kefan Zhang — 2024-02-28

Osteoarthritis is a prevalent chronic disease. One of its primary pathological processes involves the degeneration of articular cartilage. Platelet-rich plasma (PRP) contains cytokines and growth factors that can stimulate the repair and regeneration of articular cartilage tissues. PRP may also slow the progression of osteoarthritis. The purpose of this experiment is to compare the efficacy of Leukocyte poor (LP) - PRP and Leukocyte rich (LR) - PRP in treating rabbit osteoarthritis and to investigate their mechanisms of action. Analyzing the impact of leukocytes on PRP therapeutic effectiveness will provide a valuable clinical reference for the choice of which PRP is better for the treatment of osteoarthritis. A rabbit osteoarthritis model was established by injecting papain into the knee joint cavity, and LP-PRP and LR-PRP were prepared through different centrifugation methods for injection into the knee joint cavity. Eight weeks after injection, rabbit knee cartilage specimens were observed for gross changes, HE staining, senna O-solid green staining, and immunohistochemistry of type II collagen and were quantitatively compared using Pelletier's score, Mankin's pathology score, and ImageJ image processing software. Injection of papain into the knee joint cavity successfully established a rabbit model of osteoarthritis. All three evaluation indexes differed significantly from those of the blank group (P<0.05). LP-PRP and LR-PRP exhibited therapeutic effects when compared with the model group. The two PRP groups had similar gross tissue appearance and pathology (P>0.05). The LR-PRP group had higher collagen type-II expression (P < 0.05) than the LP-PRP group. Both LP-PRP and LR-PRP proved therapeutic for the rabbit papain osteoarthritis model. The difference in leukocyte content between the two groups did not yield different cartilage morphology or other factors by 8 weeks posttreatment. LR-PRP displayed the ability to release more factors relevant to the metabolism of type II collagen than LP-PRP, enabling the preservation of into cartilage collagen content of type II collagen and delaying osteoarthritis progression.

Exploring the prognostic significance of β-HCG levels in cervicovaginal secretions and maternal risk factors for early birth

Homood Alharbi — 2024-02-28

Preterm birth is one of the leading causes of death in the perinatal period, this study was conducted to investigate the predictive value of β-HCG levels in cervicovaginal secretions and maternal risk factors in preterm delivery. This cross-sectional study was conducted over a six-month period from January 1 to June 30, 2021, in Baghdad hospitals. The data were collected and used from the mothers who went to the hospital for delivery. Demographic information of patients and some risk factors were investigated. Vaginal secretions were sampled with a cotton swab. β-HCG level in weeks 29, 31, 33, and 35 was measured by ELISA method. Data were analyzed with SPSS Ver 25 software and a significance level of less than 0.05 was considered. The mean age of the study participants was 28.29 ± 5.68 years. There was a significant difference in the level of β-HCG between women with full-term delivery and pre-term women in weeks 29, 31, 33, and 35 of pregnancy (P ≤ 0.001). Maternal factors such as age older than 35 years, BMI, history of thyroid disease, blood pressure, premature rupture of the amniotic sac, parity, twin and multiple births, and decreased amniotic fluid volume have been identified as factors affecting preterm delivery. The β-HCG level can also be a helpful marker for preterm birth.

Effect of Liraglutide combined with Jinlida granules on glycolipid metabolism and islet function of type 2 diabetes mellitus

Xianjuan Wang — 2024-02-28

To investigate whether Liraglutide combined with Jinlida granules affects glycolipid metabolism and islet function in the treatment of type 2 diabetes mellitus (T2DM), a control group and an observation group were established with 90 T2DM patients. The control group was given Jinlida treatment and the observation group was given liraglutide combined treatment for 12 weeks. The clinical efficacy, glycolipid metabolism, bone metabolism, islet function, and endothelial function. The curative effect of the observation group was better than that of the control group. After treatment, FBG, 2hPG, HbAlc, TC, TG, and LDL-C in the observation group were lower and HDL-C was higher than those in the control group (P < 0.05). After treatment, the observation group showed higher bone mineral density, osteocalcin, FINS, and HOMA-β and lower HOMA-IR than the control group (P < 0.05). After treatment, endothelin-1 level in the observation group was lower than that in the control group, and the NO level was higher (P < 0.05). No significant difference was found in the incidence of adverse reactions between the two groups (P > 0.05). Liraglutide combined with Jinlida in T2DM can improve glucose, lipid, and bone metabolism, promote the recovery of islet function, and enhance vascular endothelial function.

Pathological characteristics and long-term prognosis of submucosal infiltrating carcinoma of the colon and rectum below 2 cm confirmed by endoscopic submucosal dissection

Jun Zhou — 2024-02-29

The purpose of this study was to explore the correlation between additional surgery and the clinicopathological characteristics of colorectal cancer, as well as its impact on patient prognosis. A total of 119 patients with early colorectal cancer were selected and divided into an additional surgery group (28 cases) and a non-additional surgery group (91 cases). According to the tumor size, the patients were further divided into a large diameter group (54 cases, d≥1 cm and <2 cm) and a small diameter group (65 cases, d<1 cm). The clinical and pathological characteristics as well as the prognosis of the patients were statistically analyzed. The results showed that infiltration type, depth of infiltration, and tumor size were correlated with additional surgery (P<0.05). Infiltration type and depth of infiltration were closely related to additional surgery. Differentiation degree, infiltration type, and depth of infiltration were correlated with tumor diameter. Infiltration type was closely related to tumor size. Age, depth of infiltration, and tumor size were correlated with patient survival rates. Infiltration type and depth of infiltration were closely related to patient survival rate (P<0.05). They were independent risk factors affecting patient prognosis. The 5-year disease-free survival rates were 73.33% and 72.5%, respectively, with no statistically significant difference. Infiltration type and depth of infiltration were independent risk factors for recurrence in colorectal cancer patients, while depth of infiltration was an independent risk factor for the 5-year survival rate after surgery. They can be used to predict the prognosis of colorectal cancer and guide clinical treatment as a supplement to the traditional staging of colorectal cancer.

Green synthesis and characterization of silver nanoparticles from Ducrosia flabellifolia Boiss. aqueous extract: Anti-quorum sensing screening and antimicrobial activities against ESKAPE pathogens

Mejdi Snoussi — 2024-02-28

Biosynthesis of silver nanoparticles using natural compounds derived from plant kingdom is currently used as safe and low-cost technique for nanoparticles synthesis with important abilities to inhibit multidrug resistant microorganisms (MDR). ESKAPE pathogens, especially MDR ones, are widely spread in hospital and intensive care units. In the present study, AgNPs using Ducrosia flabellifolia aqueous extract were synthesized using a reduction method. The green synthesized D. flabellifolia-AgNPs were characterized by UV–Vis spectrophotometer, Scanning electron microscopy (SEM), and X-ray diffraction assays. The tested D. flabellifolia aqueous extract was tested for its chemical composition using Liquid Chromatography-Electrospray Ionization-Mass Spectrometry (LC-ESI-MS). Anti-quorum sensing and anti-ESKAPE potential of D. flabellifolia-AgNPs was also performed. Results from LC-ESI-MS technique revealed the identification of chlorogenic acid, protocatechuic acid, ferulic acid, caffeic acid, 2,5-dihydroxybenzoic acid, and gallic acid as main phytoconstituents. Indeed, the characterization of newly synthetized D. flabellifolia-AgNPs revealed spherical shape with mean particle size about 16.961±2.914 nm. Bio-reduction of silver was confirmed by the maximum surface plasmon resonance of D. flabellifolia-AgNPs at 430 nm. Newly synthetized D. flabellifolia-AgNPs were found to possess important anti-ESKAPE activity with low minimal inhibitory concentrations (MICs) ranging from 0.078 to 0.312 mg/mL, and low minimal bactericidal concentrations (MBCs) varying from 0.312 to 0.625 mg/mL. Moreover, D. flabellifolia-AgNPs were active against Candida utilis ATCC 9255, C. tropicalis ATCC 1362, and C. albicans ATCC 20402 with high mean diameter of growth inhibition at 5 mg/mL, low MICs, and minimal fungicidal concentrations values (MFCs). The newly synthetized D. flabellifolia-AgNPs were able to inhibit violacein production in Chromobacterium violaceum, pyocyanin in Pseudomonas aeruginosa starter strains. Hence, the newly synthesized silver nanoparticles using D. flabellifolia aqueous extract can be used as an effective alternative to combat ESKAPE microorganisms. These silver nanoparticles can attenuate virulence of Gram-negative bacteria by interfering with the quorum sensing system and inhibiting cell-to-cell communication.

Tetrandrine alleviates inflammation and promotes macrophage M2 polarization in gouty arthritis by NF-κB-mediated Lcp1

Li Fang — 2024-02-28

Gouty arthritis (GA) is an inflammatory disease caused by the deposition of monosodium urate (MSU) crystals into joints. Tetrandrine (TET) is a bisbenzylisoquinoline alkaloid extracted from the root of Stephania tetrandra and can exert an anti-inflammatory function in different diseases. Nevertheless, the specific function of TET in GA remains unclear. We established the GA mouse model by MSU injection into joints of mice. Paw volume and gait score were detected for measuring the degree of joint swelling and the situation of joint dysfunction. Western blot were utilized to test the alterations of M1-related factors (IL-6, IL-1β, TNF-α, IL-12, and iNOS) and M2-related factors (Mgl1, Mgl2, Pgc1-β, Arg-1, and IL-10). The activity of NF-κB p65 in tissues was determined. The interaction of NF-κB p65 and Lcp1 was measured by ChIP and luciferase reporter assay. Lcp1 KO mice were utilized to detect the effect of Lcp1 depletion on GA process. TET treatment markedly suppressed MSU-induced joint swelling, joint dysfunction, and joint injury in GA mice. TET can also reduce inflammatory reactions in MUS-induced mice. Furthermore, we proved that TET facilitated M2 macrophage polarization and inhibited M1 macrophage polarization in GA mice. In addition, TET was found to inhibit NF-κB activity and NF-κB-mediated Lcp1 expression. Lcp1 knockdown can improve joint injury and promote M2 macrophage polarization in GA mice, while this effect was further enhanced by TET. TET alleviates inflammation and facilitates macrophage M2 polarization in GA by NF-κB-mediated Lcp1.

Association of a single-nucleotide polymorphism in C12orf43 region with the risk of coronary artery disease

Najma Qammar — 2024-02-28

The genetics of organisms play a vital role in the development of coronary artery disease (CAD), with its heritability estimated at approximately 50-60%. For this purpose, we examined the relationship between CAD risk and C12orf43/rs2258287 polymorphisms in the Pakistani population. In this study based on the genetic approach to dyslipidemia, a total of 200 subjects were included from the southern Punjab. The biochemical analysis of parameters (total cholesterol, triglycerides, blood glucose, high-density lipoprotein, and low-density lipoprotein) was carried out along with molecular analysis using an ARMS-PCR-based assay for single-nucleotide polymorphism (SNP) C12orf43/rs2258287 to identify the genotype. Genotypes showed a substantial correlation with both family history and metabolic markers. The cholesterol, low-density lipoprotein cholesterol (LDL-C), triglycerides and blood glucose levels were higher while the high-density lipoprotein cholesterol (HDL-C) level was lower significantly (p<0.05) in cases than in controls. Age, pulse rate, diabetes, physical activity, smoking, family history, and dietary habits were also significantly associated (p<0.05) with CAD individuals. The SNP C12orf43/rs2258287 also showed an association with CAD in the population of southern Punjab. Based upon this study, it could be concluded that CAD is characterized by an unfavorable lipid profile in association with SNP C12orf43/rs2258287.

Declined circular RNA mitofusin 2 constrains the deterioration of Wilms tumor via modulating microRNA-372-3p/transforming growth factor-β receptor type 2 axis

Maolin Xiao — 2024-02-28

To explore the action and mechanism in which circular RNA (circRNA) mitofusin 2 (MFN2) repressed the malignant proliferation of Wilms tumor (WT) via modulating microRNA (miR)-372-3p/transforming growth factor-β receptor type 2 (TGFBR2) axis. CircRNA MFN2 was distinctly elevated in the tissues and cells of WT patients, while miR-372-3p was silenced in the tissues and cells of WT. Test of TGFBR2, PCNA and Bax was implemented. Transfection with si-circRNA MFN2 or miR-372-3p-mimic restrained cancer cell advancement and the number of PCNA content was declined, while transfection with miR-372-3p-inhibitor was opposite, and PCNA content was augmented. MiR-372-3p-inhibitor turned around si-circRNA MFN2’s therapeutic action after co-transfection with si-circRNA MFN2 + miR-372-3p-inhibitor. Ultimately, it was verified that circRNA MFN2 was negatively associated with miR-372-3p, which was negatively linked with TGFBR2, and circRNA MFN2 was positively associated with TGFBR2. To sum up, the results of this research illuminated circRNA MFN2 repressed WT’s malignant proliferation via modulating miR-372-3p/TGFBR2 axis.

Quercetin and tanshinone prevent mitochondria from oxidation and autophagy to inhibit KGN cell apoptosis through the SIRT1/SIRT3-FOXO3a axis

Qingzhi Wan — 2024-02-28

Granulosa cells are somatic cells located inside follicles that play a crucial role in the growth and development of follicles. Quercetin and tanshinone are two key monomers in traditional Chinese medicine that have antioxidant and anti-aging properties. The KGN cell apoptosis model caused by triptolide (TP) was employed in this work to investigate granulosa cell death and medication rescue. Quercetin and tanshinone therapy suppressed KGN cell death and oxidation while also regulating the expression of critical apoptosis and oxidation-related markers such as B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax). Further research revealed that the effects of Quercetin and Tanshinone were accomplished via deacetylation of FOXO3A in the cytoplasm and mitochondria via the SIRT1/SIRT3-FOXO3a axis. In summary, Quercetin and tanshinone protect KGN cells from apoptosis by reducing mitochondrial apoptosis and oxidation via the SIRT1/SIRT3-FOXO3a axis.

Expression patterns and prognostic role of m6A RNA methylation regulators in Non-small Cell Lung Cancer

Ming Zhang — 2024-02-28

To investigate the expression pattern and prognostic role of m6A RNA methylation regulators in non-small cell lung cancer (NSCLC), we downloaded data from 422 patients from The Cancer Genome Atlas (TCGA) database. The relationship between the expression levels of m6A RNA methylation regulators and clinicopathological variables of NSCLC was analysed using R language. By analysing glioma data in TCGA, we found that a prognostic risk score model could be constructed based on 18 genes with m6A methylation modification. m6A gene alterations were significantly associated with tumour grade and tumour stage. Least Absolute Shrinkage and Selection Operator (LASSO) Cox regression models were used to identify 2 m6A RNA methylation modifiers: IFG2BP2, and METTL14 to construct risk profiles. Based on the risk profile, patients were divided into high-risk and low-risk groups. The overall survival rate of the low-risk group was significantly higher than that of the high-risk group. The results suggest that the prognostic risk score model constructed by m6A methylation regulators can predict the prognosis of glioma patients. IFG2BP2 and METTL14 may be the key m6A methylation regulators involved in the development of NSCLC and can be used as the molecular markers for the prognosis of NSCLC.

miR-127-5p regulates FAIM2-mediated cell apoptosis and participates in cerebral ischemia-reperfusion injury

Teng Hu — 2024-02-28

Mechanical thrombectomy (MT) has become an effective re-airway method for cerebral ischemia-reperfusion injury (CI/RI). However, at present, there are few studies on the impact of MT therapy on the prognosis of CI/RI patients at home and abroad. Therefore, this paper aims to analyze the relevant factors affecting the prognosis of CI/RI patients after MT therapy. The main regulatory miRNAs during CI/RI in patients with MT were screened and studied. Serums were obtained from 80 patients (moderate to severe stroke) who underwent MT. Clinical information was recorded using a unified standard questionnaire. According to the modified Rankin Scale, the patients were divided into a good prognosis group and a poor prognosis group. The clinical data were compared respectively, and univariate and multivariate Logistic regression analysis was performed. ROC curves were drawn, and Kaplan-Merier method determined whether different NIHSS scores at admission had any difference in the in-hospital survival rate of CI/RI patients treated with MT. miRNAs in serum were detected and screened out. Cell and animal models were established, in which miRNAs and apoptotic molecules were detected. miRNA target genes were predicted, and the mechanism of miRNA regulation of apoptosis was verified. Gender, smoking, drinking, diabetes, hypertension, hyperlipidemia, age, and alcohol consumption suggested no difference in the two groups. The rates of smoking history, diabetes, hypertension, and hyperlipidemia in the poor prognosis group were higher than those in the good prognosis group. Smoking and diabetes were independent risk factors for poor prognosis. miR-127-5p expression in CI/RI patients with poor prognosis was higher than that in those with good prognosis. miR-127-5p expression was also elevated in both cell and animal models. Cell apoptosis was weakened after miR-127-5p knockdown, and tissue infarction in animal models was also reduced. FAIM2 was a target gene of miR-127-5p. silencing FAIM2 enhanced apoptosis after miR-127-5p knockdown. miR-127-5p/FAIM2 axis can be a new strategy to treat and prevent brain injury in CI/RI patients treated with MT.

Genome-wide association studies of salinity tolerance in local aman rice

Nusrat Jahan — 2024-02-28

The present study aimed to identify and characterize new sources of salt tolerance among 94 rice varieties from varied geographic origins. The genotypes were divided into five groups based on their morphological characteristics at both vegetative and reproductive stages using salinity scores from the Standard Evaluation System (SES). The experiment was designed as per CRD (Completely Randomized Design) with 2 sets of salinity treatments for 8 dS/meter and 12 dS/meter, respectively compared with one non-salinized control set. Using a Soil Plant Analysis Development (SPAD) meter, assessments of the apparent chlorophyll content (greenness) of the genotypes were done to comprehend the mechanism underlying their salt tolerance. To evaluate molecular genetic diversity, a panel of 1 K RiCA SNP markers was employed. Utilizing TASSEL 5.0 software, 598 filtered SNPs were used for molecular analysis. Whole-genome association studies (GWAS) were also used to investigate panicle number per plant (pn, tiller number per plant (till), SPAD value (spad), sterility (percent) (str), plant height (ph) and panicle length (pl. It is noteworthy that these characteristics oversee conveying the visible signs of salt damage in rice. Based on genotype data, diversity analysis divided the germplasm groups into four distinct clusters (I, II, III and IV). For the traits studied, thirteen significant marker-trait associations were discovered. According to the phenotypic screening, seven germplasm genotypes namely Koijuri, Asha, Kajal, Kaliboro, Hanumanjata, Akundi and Dular, are highly tolerant to salinity stress. The greenness of these genotypes was found to be more stable over time, indicating that these genotypes are more resistant to stress. Regarding their tolerance levels, the GWAS analysis produced comparable results, supporting that salinity-tolerant genotypes having minor alleles in significant SNP positions showed more greenness during the stress period. The Manhattan plot demonstrated that at the designated significant SNP position, the highly tolerant genotypes shared common alleles. These genotypes could therefore be seen as important genomic resources for accelerating the development and release of rice varieties that are tolerant to salinity.

Astragaloside IV inhibits idiopathic pulmonary fibrosis through activation of autophagy by miR-21-mediated PTEN/PI3K/AKT/mTOR pathway

Tiaohao Li — 2024-02-28

As the main active ingredient of Astragalus, Astragaloside IV (AS-IV) can ameliorate pulmonary fibrosis. In this experiment, we studied how AS-IV reduces idiopathic pulmonary fibrosis (IPF). Bleomycin (BLM) or TGF‐β1 was treated in mice or alveolar epithelial cells to mimic IPF in vivo as well as in vitro. ASV-IV alleviated levels of inflammatory cytokines and fibrosis markers in IPF model. Through detection of autophagy-related genes, ASV-IV was observed to induce autophagy in IPF. Besides, ASV-IV inhibited miR-21 expression in IPF models, and overexpression of miR-21 could reverse the protective potential of ASV-IV on IPF. PTEN was targeted by miR-21 and was up-regulated by ASV-IV in IPF models. In addition, levels of inflammatory cytokines and fibrosis markers, autophagy, as well as the PI3K/AKT/mTOR pathway regulated by ASV-IV could be neutralized after treatment with autophagy inhibitors, miR-21 mimics, or si-PTEN. Our study demonstrates that ASV-IV inhibits IPF through activation of autophagy by miR-21-mediated PTEN/PI3K/AKT/mTOR pathway, suggesting that ASV-IV could be acted to be a promising therapeutic method for IPF.

Sema3A ameliorates the pathological progression of osteoarthritis by modulating mitochondrial damage in chondrocytes

Zhongkui Guo — 2024-02-28

Osteoarthritis (OA) is a major disease that causes disability in middle-aged and elderly people. A comprehensive understanding of its pathogenesis is of great significance in finding new clinical diagnosis and treatment schemes. The role of Semaphorin 3A (Sema3A) in OS has attracted attention recently, and the purpose of this study is to analyze the mechanisms underlying its impact on OS. First, a rat model of OS was established. Hematoxylin-eosin (HE) and TUNEL staining showed that the modeled rats presented typical pathological manifestations of OS, confirming the success of the modeling. Sema3A was significantly underexpressed in OS rats. Subsequently, Sema3A abnormal expression vectors were constructed to intervene in chondrocytes isolated from OS rats. It was found that the proliferation of chondrocytes was decreased, the apoptosis was increased, and the mitochondrial damage and autophagy were intensified after silencing Sema3A expression, while the above pathological processes were reversed when Sema3A expression was increased. In conclusion, Sema3A has an important influence on the pathological progression of OS, and molecular therapies targeting to increase Sema3A expression may become a new treatment for OS in the future.

Sirtuin 3 (SIRT3) improves sevoflurane-induced postoperative cognitive impairment by regulating mitochondrial oxidative stress

Changzong Dai — 2024-02-28

One of the most prevalent co-operative disorders is postoperative cognitive dysfunction (POCD), however, its pathogenesis remains unclear. Thus, the aim of this work was to evaluate SIRT3's impact on cognitive decline in aged mice under anesthesia. Adeno-associated virus SIRT3 vector (AAV-SIRT3) or empty vector (AAV-VEH) was injected into the hippocampal region of aged mice after sevoflurane induction in order to upregulate the expression of SIRT3. The expression levels of SIRT3, pro-inflammatory cytokines, and apoptotic factors in hippocampus tissues were identified by PCR, Western blotting, TUNEL staining, and enzyme-linked immunosorbent assay (ELISA), and the cognitive function of mice was assessed. The SIRT3 expression was down-regulated in the hippocampal tissue of anesthetized mice. SIRT3 overexpression can improve the learning and memory ability, reduce the escape latency, and increase the residence time in the platform and platform crossing ability of mice. The overexpression of SIRT3 in hippocampus can reduce the oxidative stress response and inflammatory response induced by anesthesia in mice, increase the superoxide dismutase (SOD) expression level, and decrease the expression level of MDA and inflammatory factors in hippocampus. In addition, SIRT3 overexpression can also reduce anesthetic-induced hippocampal cell apoptosis. By reducing the hippocampus mitochondrial oxidative stress response, SIRT3 plays a significant role in the pathophysiology of POCD in mice and is a potential target for POCD treatment and diagnosis.

Mitochondria-targeted reactive oxygen species blockor SS-31 blocks hepatic stellate cell activation and alleviates hepatic fibrosis by regulating NLRP3 inflammasomes

Li Liu — 2024-02-28

This study aimed to elucidate the effect of mitochondria-targeted reactive oxygen species (ROS) blockor SS-31 on hepatic stellate cells (HSC) activation during liver fibrosis. TGF-β1 was employed to induce HSC activation, while MitoSOX Red was utilized to assess the presence of mitochondrial ROS. The mitochondrial membrane potential (MMP) was measured using the JC-1 probe, and the ATP level was determined using a specific kit. The proliferation of HSCs was assessed using CCK-8 and colony formation assays, whereas flow cytometry was employed to detect HSC apoptosis. Fibrotic markers (COL1A1 and α-SMA) and NLRP3 inflammasome components (NLRP3, caspase-1, and ASC) were analyzed via Western blotting. Liver fibrosis was induced in mice using CCl4, and subsequently, histopathological changes were observed through HE staining and Masson staining. In TGF-β1-activated HSCs, mitochondrial ROS expression increased, MMP and ATP content decreased, indicating mitochondrial damage. After TGF-β1 induction, HSC proliferation increased, apoptosis decreased, and COL1A1, α-SMA, and NLRP3 inflammasome protein expression increased. After SS-31 treatment, mitochondrial ROS expression decreased, MMP recovered, ATP level increased, HSC proliferation decreased, apoptosis increased, and the expressions of COL1A1, α-SMA, and NLRP3 inflammasome decreased. NLRP3 blockor MCC950 treatment blocked HSC activation. CCL4-induced liver fibrosis mice had inflammatory cell infiltration and significant collagen fiber deposition in the liver. After SS-31 treatment, liver inflammation and collagen deposition were significantly reduced. SS-31, as a mitochondria-targeted ROS blockor, can block HSC activation by regulating the NLRP3 inflammasome, thereby alleviating liver fibrosis.

Circular RNA circ_0096041 promotes osteosarcoma cell proliferation and migration via sponging miR-556-5p and regulating LIN28A expression

Fulin Zhou — 2024-02-28

Strategies targeting lin-28 homolog A (LIN28A) for the treatment of osteosarcoma are limited, even though salient findings have illustrated the crucial role of LIN28A in bone deformities and cancer. In the present study, we proved circ_0096041, one of the circular RNAs (circRNAs) with significant upregulated expression in osteosarcoma, to be notably engaged in the progression of osteosarcoma. We elucidated that osteosarcoma patients with highly expressed circ_0096041 had relatively worse prognoses. We determined that circ_0096041 potentially sponge miR-556-5p using the Circular RNA Interactome database. Meanwhile, we proved circ_0096041 was associated with miR-556-5p. Furthermore, we determined that miR-556-5p was targeted by LIN28A directly, evidenced by in silico analysis using the miRWALK tool and in vitro analysis. Functionally, our experimental setting aimed to explore the function of circ_0096041/miR-556-5p/LIN28A axis in vitro and in vivo. Our findings demonstrated that circ_0096041 boosted the proliferation and migration of osteosarcoma via LIN28A/miR-556-5p axis. In vivo models were further established to estimate the metastasis promoted by circ_0096041. This research elucidated the enhanced osteosarcoma progression by circ_0096041 and its potential mechanism, which provided innovative targets for osteosarcoma treatment.

Effects of exosomes derived from platelet-rich plasma on osteogenic differentiation of dental pulp stem cells

Chuzi Mo — 2024-02-28

Platelet-rich plasma (PRP) can cause osteogenic differentiation of dental pulp stem cells (DPSCs). However, the effect of exosomes derived from PRP (PRP-Exos) on osteogenic differentiation of DPSCs remains unclear. Herein, we evaluated the impact of PRP-Exos on osteogenic differentiation of DPSCs. PRP-Exos were isolated and identified by transmission electron microscopy (TEM) and western blotting (WB). Immunofluorescence staining was performed to evaluate endocytosis of PRP-Exos by DPSCs. Alkaline phosphatase staining, alizarin red staining, western blot and qRT-PCR were carried out to evaluate the DPSCs osteogenic differentiation. The sequencing microRNA (miRNA) was conducted to determine the microRNA profile of PRP-Exos treated and untreated DPSCs. The results showed that endocytosis of PRP-Exos stimulated DPSCs odontogenic differentiation by elevated expression of ALP, DMP-1, OCN, and RUNX2. ALP activity and calcified nodules formation of PRP-Exos treated DPSCs were considerably elevated relative to that of the control group. MicroRNA sequencing revealed that 112 microRNAs considerably varied in PRP-Exos treated DPSCs, of which 84 were elevated and 28 were reduced. Pathway analysis suggested that genes targeted by differentially expressed (DE) miRNAs were contributed to many signaling cascades, such as the Wnt cascade. 65 genes targeted by 30 DE miRNA were contributed to Wnt signaling. Thus, it can be infered that PRP-Exos could enhance osteogenic differentiation and alter the miRNA expression profile of DPSCs.

Correlation between PD-1 and sPD-L1 expression levels in peripheral blood of DLBCL patients and their clinicopathological characteristics

Liang Wang — 2024-02-28

Molecular pathology and clinical characteristics play a crucial role in guiding treatment selection and predicting the prognosis of diffuse large B-cell lymphoma (DLBCL). The programmed cell death protein 1 (PD-1) and its ligand (PD-L1), have emerged as pivotal regulators of immune checkpoints in cancer. The objectives of this study are to investigate the correlation between the expression levels of PD-1 and soluble PD-L1 (sPD-L1) in the peripheral blood of DLBCL patients, analyze their clinicopathological characteristics, and identify the optimal beneficiary group for PD-1/PD-L1 blockade. Peripheral blood samples were collected from 36 DLBCL patients before their initial treatment at Shandong Cancer Hospital between December 2018 and July 2019. The expression levels of PD-1 and sPD-L1 were measured using flow cytometry and enzyme-linked immunosorbent assay (ELISA), respectively. The clinicopathological characteristics, including age, sex, Ann Arbor stage, International Prognostic Index (IPI) score, response to treatment, etc., were recorded for each patient. The surface expression of PD-1 on peripheral blood T cells was significantly higher in DLBCL patients compared to healthy controls. There was a significant association between elevated PD-1 expression levels and the advanced Ann Arbor stage (P=0.0153) as well as the B group (P=0.0184). Higher sPD-L1 levels were associated with the GCB subtype according to Hans's classification (P=0.0435). The expression levels of PD-1 and sPD-L1 in the peripheral blood of DLBCL patients are significantly correlated with disease stage, B group, and GCB subtype according to Hans's classification. This suggests that the PD-1/PD-L1 axis may play a critical role in specific subgroups of DLBCL. Targeting this axis could serve as a potential therapeutic strategy to enhance the clinical outcomes of DLBCL patients. Further studies are necessary to explore the prognostic implications of PD-1 and sPD-L1 expression levels in DLBCL patients.

CircSCUBE3 promoted ferroptosis to inhibit lung adenocarcinoma progression

Hongye Fu — 2024-02-28

CircRNAs can regulate ferroptosis and affect cancer development and are promising biomarkers and therapeutic targets in lung cancer. circSCUBE3 is expressed in lung adenocarcinoma (LUAD) tissues. In this study, our purpose was to study the role and regulatory mechanism of circSCUBE3 in LUAD ferroptosis. circSCUBE3 was identified to be significantly downregulated in LUAD samples and cell lines. The expression of biomarkers related to lipid oxidation (4-HNE) and ferroptosis (Ptgs2) was both downregulated in LUAD tissues, suggesting the ferroptosis resistance in LUAD. Erastin, a ferroptosis inducer, was used to stimulate the LUAD cells for 48 h. The cell viability, 4-HNE and Ptgs2 level of LUAD cells were decreased by exposure to erastin while the expression of circSCUBE3 was not significantly altered. We then overexpressed circSCUBE3 in LUAD cells and found it decreased the GSH level and GSH/GSSG ratio in LUAD cells. CircSCUBE3 might serve as an independent factor of ferroptosis and may induce ferroptosis in LUAD by inhibiting GSH synthesis. The loss-of-function experiments were conducted, and circSCUBE3 deficiency reversed the erastin-induced reduction in cell viability, GSH level, GSH/GSSG ratio, mitochondrial membrane potential and elevation in MDA content, Ptgs2, 4-HNE expression as well as lipid ROS production. CircSCUBE3 negatively regulated GPX4 expression in LUAD cells, and the silencing of GPX4 counteracted the impact of circSCUBE3 deficiency on LUAD cell viability as well as ferroptosis, suggesting that circSCUBE3 regulated the GPX4-mediated GSH synthesis in LUAD. CircSCUBE3 was to bind to CREB, which activated the transcription of GPX4. CircSCUBE3 negatively regulated GPX4 expression by competitively interacting with CREB. In the tumor-bearing mouse models, circSCUBE3 silencing promoted tumor growth and reversed the erastin treatment-induced inhibition on tumorigenesis in vivo. In conclusion, circSCUBE3 inhibited LUAD development by promoting ferroptosis via the CREB/GPX4/GSH axis, which might provide a novel option for the LUAD targeted therapy.

The effects of laparoscopic and laparotomy extensive hysterectomy on the safety of ureterovaginal fistula infection in patients with cervical cancer

Hongge Xu — 2024-02-29

This study aimed to investigate the effect of laparoscopic and laparotomy extensive hysterectomy on the safety of ureterovaginal fistula infection in patients with cervical cancer. For this purpose, a total of 90 patients with early cervical cancer admitted to Affiliated Huaian No.1 People’s Hospital of Nanjing Medical University from February 2021 to May 2022 were randomly divided into laparoscopy group and laparotomy group, with 45 cases in each group. The laparoscopy group was treated with laparoscopic extensive hysterectomy, while the laparotomy group was treated with laparotomy extensive hysterectomy. The KPS score, adverse reactions, as well as serum creatinine and urea nitrogen were compared between the two groups. Results showed that after surgery, the KPS score in both groups was higher than before treatment, and the KPS score in laparoscopy group was higher than that in laparotomy group, the difference was statistically significant (P<0.05). After operation, the incidence of adverse reactions in laparotomy group was higher than that in the laparoscopy group, the difference was statistically significant (P<0.05). Moreover, after operation, the levels of creatinine and urea nitrogen in laparoscopy group were significantly lower than those in laparotomy group, the differences were statistically significant (P<0.05). In conclusion, both laparoscopic and laparotomy extensive hysterectomy may lead to ureterovaginal fistula infection in patients with cervical cancer. However, compared with laparotomy extensive hysterectomy, laparoscopic extensive hysterectomy had higher safety and significantly improved the quality of life of patients, which was worthy of popularization and application in clinical practice.