The aim of present study was to evaluate the potential effects of Rhodiola crenulata oral liquid (RCOL) on exhaustive exercise (EE)-induced fatigue in mice. Male Institute of Cancer Research mice from five treatment groups (n=10 per group) were orally administered with sterilized water for the Control and EE groups and/or RCOL at doses of 1.02, 3.03 and 6.06 ml/kg/day, once daily for 2 weeks. Anti-fatigue activity was subsequently evaluated by measuring the levels of creatine kinase (CK), lactic acid (LA), lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and total anti-oxidative capability (T-AOC). Histopathology was assessed using hematoxylin and eosin staining. Ultrastructures of mitochondria were observed by transmission electron microscopy. Energy supply capacity was assessed using citrate synthase (CS), succinate dehydrogenase (SDH), Na+-K+-ATPase, and liver and quadriceps glycogen content assays. Expression levels of mRNA and protein associated with mitophagy in the skeletal muscle were measured by reverse transcription-quantitative PCR and western blotting, respectively. RCOL was observed to markedly inhibit fatigue-induced oxidative stress by increasing the activities of SOD, CAT and T-AOC, whilst reducing the accumulation of LA, CK, LDH and MDA. Histological analysis of the quadriceps femoris tissue suggested increased numbers of muscle fibers in the RCOL groups compared with those in the EE group. RCOL administration was found to reverse EE-induced mitochondrial structural damage and alleviated defects inflicted onto the energy supply mechanism by increasing CS, SDH, Na+-K+-ATPase and glycogen levels. Additionally, RCOL reduced the protein expression of PTEN-induced kinase 1 (PINK1), Parkin, microtubule-associated proteins 1A/1B light chain 3, sequestosome 1 and ubiquitin, whilst lowering the gene expression of PINK1 and Parkin. Taken together, results from the present study clarified the anti-fatigue effect of RCOL, where the underlying mechanism may be associated with increased antioxidant activity, enhanced energy production and the inhibition of mitophagy by suppressing the PINK1/Parkin signaling pathway.

The purpose of the present study was to investigate the effect of microRNA (miR)-144-5p on human umbilical vein endothelial cells (HUVECs) to explore the role of miR-144-5p in atherosclerosis. miR-144-5p expression was upregulated in HUVECs using miR-144-5p mimics. The relative expression level of miR-144-5p in HUVECs was detected using reverse transcription-quantitative PCR (RT-qPCR). Cell proliferation was detected by performing an MTT assay. Apoptosis was determined via flow cytometry. Cell migration ability was detected by a wound-healing assay. Cell invasion was determined by a transwell assay. The protein levels of phosphorylated (p)-PI3K, p-Akt and endothelial nitric oxide synthase (eNOS) were detected using western blot analysis. The binding sites between miR-144-5p and 3'-untranslated region of rapamycin-insensitive companion of mTOR (RICTOR) mRNA were predicted by TargetScan and confirmed by a dual luciferase reporter assay. The present study showed that miR-144-5p mimics significantly inhibited cell proliferation and induced apoptosis in HUVECs. In addition, miR-144-5p mimics could suppress migration and invasion of HUVECs. Further analysis identified that RICTOR was a direct target gene of miR-144-5p. Moreover, miR-144-5p upregulation decreased the protein level of p-PI3K, p-Akt and eNOS. In conclusion, miR-144-5p regulated HUVEC proliferation, migration, invasion, and apoptosis through affecting the PI3K-Akt-eNOS signaling pathway by altering the expression of RICTOR. These results indicated that miR-144-5p may be a potential target for the prevention and treatment of atherosclerosis.

Apocynum venetum is a traditional medicine that is rich in polyphenols. Apocynum venetum polyphenol extract (AVP) contains the active substances neochlorogenic acid, chlorogenic acid, rutin, isoquercitrin, astragaloside and rosmarinic acid. In the present study, the preventive effect of AVP against D-galactose-induced oxidative stress was studied in a mouse model. The sera, skin, livers and spleens of mice were examined using hematoxylin and eosin staining, reverse transcription-quantitative PCR and western blot analysis. The biochemical results showed that AVP improved the thymus, brain, heart, liver, spleen and kidney indices in a mouse model of oxidative stress. AVP was also able to reverse the reduction in levels of superoxide dismutase (SOD), glutathione peroxidase and glutathione, and increased the levels of nitric oxide and malondialdehyde identified in the serum, liver, spleen and brain of mice exposed to oxidative stress. Pathological observations confirmed that AVP could inhibit oxidative damage to the skin, liver and spleen of mice caused by D-galactose. Further molecular biological experiments also demonstrated that AVP increased the expression of neuronal nitric oxide synthase, endothelial nitric oxide synthase, Cu/Zn-SOD, Mn-SOD, catalase, heme oxygenase-1, nuclear factor-erythroid 2-related factor 2, γ-glutamylcysteine synthetase and NAD(P)H quinone dehydrogenase 1 and reduced the expression of inducible nitric oxide synthase in the liver and spleen of treated mice compared to controls. Notably, the preventive effect of AVP against D-galactose-induced oxidative damage in mice was better than that of the confirmed antioxidant vitamin C. In conclusion, AVP exhibited an antioxidant effect and the AVP-rich Apocynum venetum may be considered a plant resource with potential antioxidative benefits.

Discs large-associated protein 5 (DLGAP5) is a microtubule-associated protein and is reported to exert oncogenic role in tumorigenesis, including lung cancer and hepatocellular carcinoma. However, the prognostic value and biological function of DLGAP5 in ovarian cancer (OC) still remain unclear. The present study investigated the expression pattern of DLGAP5 by searching the Oncomine microarray database. The correlation between DLGAP5 and survival prognosis of OC patients was analyzed by the online tool KM-plotter. Knockdown of DLGAP5 was achieved by transfection with small interfering RNA targeting DLGAP5 in two OC cell lines (SKOV3 and CAOV3). Cell proliferation was assessed by Cell Counting Kit-8 assay and colony-formation assay. Flow cytometry was utilized to determine the effects of DLGAP5 on cell cycle distribution and apoptosis. The present study data showed that DLGAP5 was significantly upregulated in OC and its higher expression was associated with poor survival prognosis. Knockdown of DLGAP5 significantly suppressed cell proliferation, induced cell cycle G2/M phase arrest and apoptosis. Western blot analysis further demonstrated that DLGAP5 knockdown downregulated the expression of CDK1, Cyclin B1 and Bcl-2, but upregulated Bax expression. Collectively, these data demonstrate that DLGAP5 might be a promising prognostic therapeutic target for OC treatment.

Previous studies have indicated that microRNA (miR)-210-3p is upregulated in NSCLC, however, the specific mechanism underlying the role of miR-210-3p in NSCLC pathogenesis requires further investigation. The aim of the present study was to explore the roles of miR-210-3p in NSCLC and the associated mechanisms. A total of 30 NSCLC tissues and paired adjacent normal tissues were collected for study. Reverse transcription-quantitative polymerase chain reaction was performed to compare the expression of miR-210-3p in the 30 paired cancerous and adjacent normal tissues. Additionally, the expression of miR-210-3p in different NSCLC lines and normal human lung epithelial cell line BEAS-2B were also compared. Furthermore, A549 and H1299 NSCLC cells were cultured and transfected with miR-210-3p inhibitors, and MTT and propidium iodide/annexin V assays were performed to investigate the effects of miR-210-3p inhibition on the proliferation and apoptosis of the cells. RT-qPCR and western blot analyses were also performed to determine the effects of miR-210-3p on the expression levels of SIN3A, B-cell lymphoma 2 (Bcl-2) and Caspase-3. Finally, a reverse experiment was conducted by transfecting A549 cells with miR-210-3p inhibitor and SIN3A small interfering (si)RNA, and a dual-luciferase reporter assay was performed to confirm that SIN3A is a direct target of miR-210-3p. It was observed that miR-210-3p was significantly upregulated in NSCLC tissues compared with the levels in the adjacent normal tissues, and that the expression of miR-210-3p in patients with NSCLC was negatively correlated with the expression of SIN3A in NSCLC tissue. miR-210-3p was also significantly upregulated in different NSCLC cell lines compared with the levels in BEAS-2B cells. The transient downregulation of miR-210-3p in A549 cells led to a significant suppression of cell proliferation and markedly increased cell apoptosis, as well as increased the expression of SIN3A and Caspase-3 and decreased the expression of Bcl-2. On the other hand, co-transfection of miR-210-3p inhibitor and SIN3A siRNA partially blocked miR-210-3p inhibitor-induced pro-apoptotic effects. The results of the dual-luciferase reporter assay demonstrated that SIN3A is a direct target of miR-210-3p. Collectively, these findings indicate that can regulate the proliferation and apoptosis of NSCLC cells by targeting SIN3A. These results suggest that miR-210-3p has the potential to become a novel therapeutic target for the treatment of NSCLC.

Oral squamous cell carcinoma (OSCC), one of the most common types of human cancer, has a high mortality rate and a poor prognosis due to its high rates of recurrence and metastasis. In recent years, icariin (ICA) has been reported to play an important role in a variety of malignancies, such as gastric, colorectal, pancreatic and ovarian cancer. However, its role and mechanism in OSCC remains to be elucidated. The present study aimed to investigate the effect of ICA in OSCC cells and to reveal its underlying mechanisms. The OSCC cell lines SCC9 and Cal 27 were used to explore the effect of different concentrations of ICA on the biological behavior of OSCC cells. The effect of ICA on OSCC cell proliferation and apoptosis was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide and flow cytometric assays, respectively. Subsequently, the protein expression levels of caspase-3 and cleaved-caspase-3 were detected using western blot analysis. Additionally, the protein and mRNA expression levels of nuclear factor-κB (NF-κB) and phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT) signaling pathway-related factors were determined using western blot analysis and reverse transcription-quantitative PCR, respectively. The results demonstrated that ICA inhibited OSCC cell proliferation and significantly increased the apoptosis rate in a dose-dependent manner. In addition, treatment of OSCC cells with ICA upregulated the protein expression of cleaved-caspase-3 and increased the cleaved-caspase-3/caspase-3 ratio. The protein expression levels of phosphorylated (p)-p65, p-PI3K and p-AKT were decreased in OSCC cells treated with ICA. The aforementioned findings revealed that ICA could attenuate the proliferation of OSCC cells and induce apoptosis via inhibiting the NF-κB and PI3K/AKT signaling pathways. Therefore, the current study provided a new insight into the clinical treatment of OSCC.

Abnormal activation of the Wnt signaling pathway is found in 90% of colorectal cancers (CRCs). Secreted frizzled-related protein 4 (sFRP4) serves as an antagonist of the canonical Wnt signaling pathway. Epigenetic alterations, including changes in DNA methylation and histone methylation, may influence the expression of sFRP4. Polycomb group (PcG) proteins are epigenetic transcriptional repressors that selectively repress gene expression by forming polycomb repressive complexes (PRCs). Enhancer of zeste homolog 2 (EZH2), the core component of PRC2, is a histone-lysine N-methyltransferase that interacts with DNA methyltransferases. In the present study, the promoter DNA methylation status of sFRP4 in CRC cell lines was analyzed and the underlying mechanisms of action governing this modification was investigated. Firstly, the DNA methylation status of the sFRP4 promoter in CRC cell lines was assessed using methylation-specific PCR. Subsequently, the mRNA and protein levels of sFRP4 were measured using real-time qPCR and western blot analysis, respectively, to determine whether the DNA methylation status of the sFRP4 promoter is correlated with its transcriptional levels. To screen for important epigenetic modifiers that may regulate the promoter DNA methylation status of sFRP4, the expression levels of PcG proteins were examined by gene array analysis. ChIP-qPCR was performed to test whether the selected PcG proteins directly bind the promoter region of sFRP4. Finally, the downregulated PcG proteins EZH2, chromobox 7 (CBX7) and jumonji and AT-rich interaction domain containing 2 (JARID2) were identified and their association with sFRP4 expression levels and Wnt/β-catenin signaling pathway activity were investigated. The present study revealed that sFRP4 was hypermethylated in the promoter region and downregulated during the progression of the CRC cell lines from Dukes A to Dukes C. Expression levels of PcG proteins EZH2, CBX7 and JARID2 were upregulated and positively associated with the aberrantly activated Wnt signaling pathway in the CRC cell lines. EZH2, CBX7 and JARID2 were all enriched in the sFRP4 promoter region in CRC cells. EZH2 downregulation did not affect the promoter DNA methylation status of sFRP4 but increased its expression levels and decreased CRC cell proliferation. DNA methylation controls the expression of sFRP4. EZH2 regulates sFRP4 expression without affecting the DNA hypermethylation of the sFRP4 promoter and influences CRC cell proliferation and Wnt/β-catenin signaling pathway activities.

The aim of the present study was to investigate the potential changes in the choroidal thickness (CT) after surgical implantation of collamer lens (ICL) and to determine whether the variations in CT were associated with the degree of myopia. In the study, 98 eyes from 98 myopia patients were divided into two groups according to the degree of myopia: High myopia and super-high myopia. All eyes were measured using the swept-source optical coherence tomography (SS-OCT) technique. CT and CT variations were also recorded. The foveal CT increased significantly in high-myopia patients at 2 h after surgery and 3 months after surgery; the same tendency was observed in the inner nasal CT and outer nasal CT at the same time-points. In patients with super-high myopia, the subfoveal CT increased significantly at 2 h and 3 months after surgery compared with the pre-operative values. No statistically significant differences were obtained in any of the nine different choroidal regions evaluated at post-operative week 1 and post-operative month one. Furthermore, the increase in the subfoveal CT in the super-myopia group was significantly higher than that in the high-myopia group at 2 h and at 3 months after ICL. The results of the present study indicated that the CT significantly increased 2 h after the surgery and then reached a peak at 3 months, particularly in the subfoveal and nasal areas. A higher degree of myopia was associated with greater subfoveal choroidal changes.

Mesenchymal stem cells (MSC) are able to differentiate into cells of multiple lineage, and additionally act to modulate the immune response. Interleukin (IL)-9 is primarily produced by cluster of differentiation (CD)4+ T cells to regulate the immune response. The present study aimed to investigate the effect of human umbilical cord derived-MSC (UC-MSC) on IL-9 production of human CD4+ T cells. It was demonstrated that the addition of UC-MSC to the culture of CD4+ T cells significantly enhanced IL-9 production by CD4+ T cells. Transwell experiments suggested that UC-MSC promotion of IL-9 production by CD4+ T cells was dependent on cell-cell contact. Upregulated expression of CD106 was observed in UC-MSC co-cultured with CD4+ T cells, and the addition of a blocking antibody of CD106 significantly impaired the ability of UC-MSC to promote IL-9 production by CD4+ T cells. Therefore, the results of the present study demonstrated that UC-MSC promoted the generation of IL-9 producing cells, which may be mediated, in part by CD106. The findings may act to expand understanding and knowledge of the immune modulatory role of UC-MSC.

The aim of the present study was to explore the effect of baicalin on liver hypoxia/reoxygenation (H/R) injury and the possible mechanism involved. A cellular H/R model was established and cells were treated with 50, 100 and 200 µmol/l baicalin. Following reoxygenation for 6 h, cell viability, lactate dehydrogenase (LDH), B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), caspase 3 and cleaved caspase 3 were assessed. Furthermore, levels of endoplasmic reticulum stress markers binding of immunoglobulin protein (BIP) and CCAAT/enhancer-binding protein homologous protein (CHOP) and autophagy markers microtubule-associated proteins 1A/1B light chain 3B (LC3) and beclin 1 were measured. To confirm the involvement of autophagy in baicalin-mediated attenuation of H/R injury, the autophagy inhibitor 3-methyladenine (3-MA) was administered. The results revealed that baicalin administration increased cell viability and decreased LDH levels, most notably at a dosage of 100 µmol/l. Baicalin pretreatment also downregulated the expression of caspase 3, cleaved caspase 3 and Bax, while upregulating the expression of Bcl-2. Furthermore, BIP and CHOP were decreased while LC3 and beclin-1 were significantly increased by baicalin pretreatment. Inhibiting autophagy using 3-MA, resulted in a significant decrease in LC3-II, beclin-1 and LDH, as well as increase in the expression of BIP, CHOP, caspase 3, cleaved caspase 3 and Bax. Bcl-2 and cell viability were also decreased. In conclusion, the results of the present study indicate that baicalin exerts a protective effect on liver H/R injury and this may be achieved via the induction of autophagy.

Cervical cancer (CC) is one of the most malignant tumors that affect women. Recent studies have reported that microRNAs (miRs) serve important roles in CC. The aim of the present study was to investigate the role of miR-218 in CC and to verify its underlying mechanism. The results of reverse transcription-quantitative polymerase chain reaction (RT-qPCR) revealed that miR-218 was dramatically downregulated in CC tissues and cell lines. Furthermore, the expression of Gli3 and Ki67 was measured using RT-qPCR and the results revealed that levels of these proteins were negatively correlated with miR-218 in CC tissues. The protein expression levels were determined by western blotting. Then SiHa cell line was used to investigate the mechanism of CC. Following cell transfection, cell apoptosis and cycle analyses were performed using the flow cytometry. The results revealed that miR-218 overexpression significantly inhibited cell proliferation, apoptosis and cell cycle. Additionally, luciferase reporter assay revealed that Gli3 may be a novel and direct target of miR-218 in CC. Taken together, the results of the present study suggest that miR-218 overexpression or Gli3 knockdown may have potential as therapeutic strategies for the treatment of CC.

Atherosclerosis is a multifactorial chronic disease that is a major cause of death and injury worldwide. Apoptosis of endothelial cells (ECs) serves an important role in the occurrence and development of atherosclerosis. MicroRNAs (miRNAs) serve a key role in atherosclerosis though regulating the function of ECs. At present, the role of miRNA-144-5p (miR-144-5p) in atherosclerosis is unclear. The aim of this study was to investigate the effect of miR-144-5p on atherosclerosis in oxidized low-density lipoprotein (ox-LDL)-stimulated human umbilical vein endothelial cells (HUVECs). Results from the present study demonstrated that miR-144-5p overexpression could inhibit proliferation and induce apoptosis in HUVECs. To further study the biological function of miR-144-5p, the effects of modulating miR-144-5p expression on the invasion and migration of HUVECs were also examined. The results demonstrated that miR-144-5p upregulation suppressed HUVEC migration and invasion. TargetScan and dual luciferase reporter assay results demonstrated that SMAD1 was a direct target gene of miR-144-5p. miR-144-5p upregulation inhibited the expression of phosphorylated-SMAD1/5/8 in the SMAD pathway. In conclusion, the data indicated that miR-144-5p serves an important role in the development of atherosclerosis through regulating the function of HUVECs by targeting SMAD1.

This study investigated the correlation between interleukin (IL)-1β-511C/T gene polymorphism and myocardial infarction (MI) complicated with ischemic stroke (IS). A total of 251 MI patients complicated with IS (observation group) and 200 healthy people (control group) were selected for the case-control study. IL-1β-511C/T gene polymorphism was detected via polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The genotype distribution and allele frequency were compared between the two groups, and the correlation between gene polymorphism and MI complicated with IS, was analyzed after traditional risk factors were adjusted by using logistic regression method. The frequencies of CT and TT genotypes in the observation group were higher than those in the control group (P<0.05). The frequency of T allele in the observation group was significantly higher than that in the control group (P<0.05), but the frequency of C allele was obviously lower than that in the control group (P<0.05). According to results of logistic regression analysis, arrhythmia and high-density lipoprotein cholesterol (HDL-C) were associated with MI complicated with IS. In patients with arrhythmia, the risk of disease in carriers with IL-1β-511T gene was 1.7-1.8 times that in non-carriers [odds ratio (OR) = 1.742 and 1.839, P<0.05]. In patients with abnormal HDL-C, the risk of disease in carriers with IL-1β-511T gene was 2.0-2.2 times that in non-carriers (OR = 2.011 and 2.249, P<0.05). Besides, the risk of MI complicated with IS in carriers with CC genotype had no significant difference in patients with arrhythmia and abnormal HDL-C (P>0.05). IL-1β-511C/T gene polymorphism may be related to the risk of MI complicated with IS.

The present study aimed to investigate the effect of the inhibition of miR-145 on cyclin-dependent protein kinase 6 (CDK6) and the proliferation of human cervical carcinoma cells. The miR-145 sequence was synthesized and cloned into pcDNA™6.2-GW to construct the recombinant plasmid pcDNA6.2-GW-miR-145. HeLa cells were divided into the micro (mi)R-145, normal control and blank groups. The transcription levels of miR-145 and CDK6 were detected using quantitative polymerase chin reaction and western blot analysis was used to examine the CDK6 protein expression. In addition, the inhibitory effect of miR-145 on the proliferation of HeLa cells was measured by an MTT assay. The recombinant plasmid pcDNA6.2-GW-miR-145 was successfully constructed and used to transfect the HeLa cells in the MiR-145 group. The miR-145 expression level in the miR-145 group was significantly higher than that in the blank group. The CDK6 expression level in miR-145 group was significantly lower than that in the blank group. Furthermore, miR-145 inhibited the proliferation of HeLa cells. In conclusion, miR-145 overexpression suppresses the expression of CDK6 and inhibits the proliferative ability of HeLa cells.

Response gene to complement (RGC)-32 regulates the cell cycle in response to complement activation. The present study demonstrated that the expression level of RGC-32 is higher in human non-small-cell lung cancer (NSCLC) tissues compared with health controls. Overexpressing RGC-32 induced p65 nucleus translocation, significantly increased nuclear p65 levels and promoted the proliferation of A549 cells. Knockdown of RGC-32 by short hairpin RNA decreased the expression level of nuclear p65 and inhibited cell proliferation. The increase in cell proliferation induced by RGC32 could be abolished by the NF-κB inhibitor pyrrolidine dithiocarbamate. Mechanistic studies indicated that RGC32 mediated NF-κB downstream genes, including vascular cell adhesion protein 1, interleukin-6, cyclin dependent kinase inhibitor 2C, testin and vascular endothelial growth factor A. In summary, the present study demonstrated a novel role of RGC-32 in the progression of NSCLC via the NF-κB pathway and p65. Therefore, RGC-32 could be a potential therapeutic target for NSCLC.