Objective To investigate the expression of biliverdin reductase (BVR) in bone marrow derived macrophages and RAW264. 7 cells. Methods L929 cells were cultured for 2 days, the supernatant was then collected. The murine bone marrow derived macrophage cells were isolated and were cultured with the above supernatant for 7 days. Cells were then divided into three groups: control group, LPS + IFN-γ group, IL-4 + IL-13 group. Cells in the LPS + IFN-γ group and the IL-4 + IL-13 group were stimulated by 100 ng / mL LPS and IFN-γ or 10 ng / mL IL-4 and 10 ng / mL IL-13 for 24 hours respectively. Western blotting was used to detect the expression levels of INOS and ARG-1 proteins. RT-PCR was used to detect the RNA expression levels of INOS, ARG-1, FN and FIZZ1. Flow cytometry was used to detect the expression of CD45, F4 / 80, CD11b, CD86 and DECTIN-1 on cells. . The expression level of BVR in the macrophage polarization model established in BMDM cells and RAW264. 7 cells was detected by RT-PCR. Results ① The protein expression level of INOS was increased in the LPS + IFN-γ group, and the protein expression level of ARG-1 was increased in the IL-4 + IL-13 group. ② The expression of CD86 on cells was increased in the LPS + IFN-γ group, and the expression of DECTIN-1 on cells was increased in the IL-4 + IL-13 group. ③ The RNA expression levels of INOS and TNF-α were increased in the LPS + IFN-γ group, the RNA expression levels of ARG-1, FN and FIZZ1 were increased in IL-4 + IL-13 group. ④ The expression level of BVR was increased in the M2-shift macrophages, but decreased in the M1-shift macrophages in BMDM and RAW264. 7 cell models. Conclusion ① The macrophage polarization models are successfully established by stimulation of LPS and IFN-γ or IL-4 and IL-13 in vitro. ② The expression level of BVR is increased in the M2-shift macrophages, but decreased in the M1- shift macrophages in vitro.

Objective To explore the effects of miR-485 on cerebral ischemia-reperfusion (CI/ R) injury and oxygen glucose deprivation / resupply ( OGD/ R) induced apoptosis in BV2 cells. Methods The CI/ R injury rat model was established by thread embolism method, rats were then injected with agomir-485 through caudal vein to observe their brain injury at 24 h time point after reperfusion. miR-485 overexpressed BV2 cell line was constructed, cells were then induced with OGD/ R to observe cells apoptosis and oxidative stress in vitro. Results The results of in vivo experiment showed that the expression level of miR-485 in brain tissues of CI/ R rats was decreased, and agomir-485 intervention could alleviate the neurological damage, reduce the level of Bax / Bcl-2 and the activities of LDH and GSH-px, and increase the activity of SOD in CI/ R rats. The results of in vitro experiment showed that OGD induction could reduce the expression level of miR-485. The overexpression of miR-485 could increase SOD activity, reduce apoptosis rate and the activities of LDH and GSH-px. Conclusion Overexpression of miR-485 can relieve CI/ R injury and reduce the OGD/ R induced apoptosis in BV2 cells, the mechanism may be related to the regulation of oxidative stress.

Objective To explore the role and potential mechanism of miR-27b-3p in the proliferation, apoptosis and cisplatin (DDP) resistance of oral squamous cell carcinoma (OSCC) resistant cells. Methods RT-qPCR was applied to detect the expression of miR-27b-3p and HOXB8 mRNA. Western blotting was applied to detect the expression of HOXB8 and drug resistance protein MRP1. CCK-8 assay was conducted to measure the cell viability, colony formation assay was used to analyze cell proliferation, and flow cytometry to detect apoptosis. The interaction between miR-27b-3p and HOXB8 was predicted by the online software Targetscan and confirmed by dual-luciferase reporter gene assay. A nude mouse xenograft model was used to test the role of miR-27b-3p in OSCC DDP resistance in vivo. Results The expression level of miR-27b-3p was obviously lower in DDP resistant OSCC tissues and OSCC cell lines, and the expression level in DDP-resistant OSCC cell lines (CAL-27 / DDP) was obviously lower than that in normal OSCC cell lines (CAL-27) (P < 0. 05). HOXB8 mRNA, by contrast, was highly expressed in DDP-resistant OSCC tissues and OSCC cell lines, and the expression in DDP-resistant OSCC cell line (CAL-27 / DDP) was obviously higher than that in normal OSCC cell line (CAL-27) (P < 0. 05). miR-27b-3p mimic could obviously inhibit the cell viability, colony formation ability and the expression of drug resistance protein MRP1 in CAL-27 / DDP cells, and promote the cell apoptosis (P < 0. 05) . Overexpression of HOXB8 could partially reverse the effects of miR-27b-3p mimics on the proliferation, apoptosis and drug resistance in CAL-27 / DDP cells (P< 0. 05). HOXB8 had a targeted regulatory role with miR-27b-3p. Intratumoral injection of miR-27b-3p agomir obviously alleviated the growth of resistant cells in the xenograft in vivo, and reduce the expression level of HOXB8 and MRP1 (P< 0. 05). Conclusion miR-27b-3p may regulate the proliferation, apoptosis and DDP resistance of OSCC drug resistant cells by targeting HOXB8.

Objective To study the protective effect of ropivacaine on cerebral ischemia / reperfusion (CI/ R) rat brain injury and hippocampal neuronal apoptosis and its mechanism. Methods Sprague-Dawley (SD) rats were divided into 5 groups: control group, CI/ R model group, ropivacaine-low ( 0. 5 mg / kg) group, ropivacaine-medium ( 1 mg / kg) group, and ropivacaine-high (2 mg / kg) group. Step-down test and Y maze test were used to evaluate the behavior of rats in each group. Longa method was used to evaluate the neurological damage in rats. HE staining was used to observe the pathological damage in brain tissue. TUNEL staining was used to detect the apoptosis in hippocampal tissue. Western blotting was used to detect the expression of associated proteins. Results Compared with the control group, the number of errors in step-down test was significantly increased in the CI/ R group, the number of new arm entry were significantly decreased, and the brain index and neurological deficit score were significantly increased. The brain tissue showed obvious pathological damage, and apoptosis was observed in neurons in the hippocampus. The expression levels of BDNF and NGF proteins were significantly decreased, Bax / Bcl-2 and cleaved Caspase-3 / Caspase-3 ratios were significantly increased, and the phosphorylation levels of P38MAPK and NF-κB P65 were significantly increased. Compared with the CI/ R group, ropivacaine treatment improved the behavioral biases caused by CI/ R injury, significantly reduced the brain index and the neurological deficit score. Ropivacaine alleviated brain tissue injury and hippocampus neuron apoptosis. Ropivacaine suppressed the activation of P38MAPK/ NF-κB P65 pathway. Conclusion Ropivacaine treatment can alleviate brain injury and hippocampal neuronal apoptosis in CI/ R rats, and its underlying mechanism may be related to the inhibition of the activation of the MAPK pathway. 

Objective To investigate the effect of long non-coding RNA (lncRNA) OSTN antisense RNA1 (OSTN-AS1) on the proliferation, migration and invasion of prostate cancer LNCaP cells and its mechanism. Methods Using prostate epithelial cell RWPE-1 as control, the expression of OSTN-AS1 and miR-4461 in LNCaP cells were detected by real-time fluorescence quantitative PCR (qRT-PCR). LNCaP cells were transfected with OSTN-AS1 small interfering RNA ( si-OSTN-AS1) or miR-4461 mimic, or co-transfected with si-OSTN-AS1 and miR-4461 inhibitor, and then cell proliferation, migration, invasion and related proteins [Ki-67, matrix metalloproteinase (MMP) -2, MMP-9] expression were detected by cell counting Kit-8 ( CCK-8) method, scratch assay, transwell and Western blotting, respectively. The dual-luciferase reporter gene experiment was used to verify the regulatory relationship between OSTN-AS1 and miR-4461. Results The expression level of OSTN-AS1 in the LNCaP cells was higher than that in the RWPE-1 cells (3. 38 ± 0. 26 vs. 1. 00 ± 0. 00, P < 0. 05), but the expression level of miR-4461 was lower than that in the RWPE-1 cells (0. 46 ± 0. 04 vs. 1. 00 ± 0. 00, P < 0. 05). After interfering with OSTN-AS1 siRNA or miR-4461 mimic, the proliferation activity, wound healing rate, and invasion number of LNCaP cells were decreased, and the expression levels of Ki-67, MMP-2, and MMP-9 were also decreased (all P < 0. 05). OSTN-AS1 targeted and bound to miR-4461, and the expression level of miR-4461 was increased in LNCaP cells that interfered with OSTN-AS1 (P< 0. 05). Down-regulation of miR-4461 reversed the effect of si-OSTN-AS1 on LNCaP cells proliferation, migration, invasion and related protein expression. Conclusion Interfering with OSTN-AS1 can inhibit the proliferation, migration and invasion of prostate cancer LNCaP cells by targetly up-regulation of miR-4461.

Objective To study the expression level of microRNA-340 (miR-340) in human glioma and its relationship with the expression of IDH1, p53, CD34 and Ki-67 proteins, and the effect of miR-340 on the expression of glucose transporter 1 / 6 (GLUT1 / 6) in glioma. Methods The expression level of miR-340 was detected by qRT-PCR. Cells were transfected with miR-340 mimics or inhibitor, the expression level of GLUT1 and GLUT6 RNA was detected by qRT-PCR, and the expression level of GLUT1 and GLUT6 protein was detected by Western blotting. Results The expression levels of miR-340 in the glioma tissues and glioma cell lines were significantly lower than those in the normal brain tissues and the normal human glial cell line HEB, respectively (P< 0. 001). The expression level of miR-340 in glioma samples from patients was negatively correlated with the Ki-67 expression level (P = 0. 0012). miR-340 mimics overexpression significantly inhibited the cell proliferation, increased the apoptosis, and reduced the migration and invasion in U87 and U251 cells. Meanwhile, miR-340 mimics significantly inhibited the expression of GLUT1 and GLUT6 in the glioma cell lines (P< 0. 001). Conclusion miR-340 is lowly expressed in the glioma tissues and cells, and the expression level of miR-340 is negatively associated with the expression level of Ki-67. miR-340 affects the glucose metabolism in glioma cells by reducing the expression levels of GLUT1 and GLUT6, which inhibits the cell proliferation and reduces the malignancy of glioma cells. miR-340 can be used as a biomarker for the early diagnosis, treatment effect evaluation and prognosis prediction of glioma. 

Objective To investigate the protective effect of glycyrrhizic acid on neurons in rats with cerebral ischemia / reperfusion. Methods An cerebral ischemia-reperfusion model was constructed in rat primary neurons in vitro by oxygen glucose deprivation reperfusion (OGD/ R) method. The changes of cell morphology were observed by microscope, and the expression of neuron-specific enolase ( NSE) was observed by immunofluorescence staining. The damaged neurons were treated with glycyrrhizic acid, and the CCK-8 method was performed to detect the viability of neuronal cells, and the level of LDH released from neurons was detected by using the lactate dehydrogenase (LDH) -cytotoxicity assay kit. The AMPK activator AICAR was used to activate the AMPK/ mTOR pathway in neurons, and the Western blotting assay was used to detect the expression levels of AMPK/ mTOR pathway-related proteins and autophagy-related proteins. Results Glycyrrhizic acid inhibited the OGD/ R-induced enhancement of neuronal LDH level and the expression levels of autophagy-related proteins Beclin 1 and LC3-Ⅱ, and the suppression of cell viability. The OGD/ R induced activation of AMPK/ mTOR pathway was also suppressed after glycyrrhizic acid treatment. Conclusion Glycyrrhizic acid exerts a protective effect on neurons in rats with cerebral ischemia-reperfusion by inhibiting neuronal autophagy through AMPK/ mTOR pathway.

Objective To investigate the effect of obesity-associated protein (FTO) and F-box and WD-40 domain protein 7 (FBXW7) on cervical squamous cell carcinoma (CSCC) radiotherapy and chemotherapy resistance. Methods Real-time polymerase chain reaction (RT-PCR) and Western blotting were used to detect the expression levels of FTO and FBXW7 in CSCC cells. The TCGA (The Cancer Genome Atlas) data were used to analysis FTO and FBXW7 expression and correlation. FTO was overexpressed in CSCC cells, and immunoprecipitation was used to detect the changes in m6A expression of FBXW7. The MTT method was used to detect the role of FTO and FBXW7 in the resistance of CSCC cells to radiotherapy and chemotherapy. Results Compared with the human normal cervical epithelial cells (HUCEC), FTO and FBXW7 were weakly expressed in CSCC cells (SiHa, c-33a) (P < 0. 05). FTO regulated the m6A of FBXW7 mRNA. The cell survival ratio was decreased after the FTO-overexpressed CSCC cells were irradiated or treated with cisplatin (P< 0. 05). The inhibition of FBXW7 increased the cell survival ratio in irradiated or cisplatin treated FTO-overexpressed CSCC cells, which induced CSCC Chemo-Radiotherapy (P < 0. 05). Conclusion FTO could remove the m6A methylation modification of FBXW7, up-regulate the expression of FBXW7, and inhibit the resistance of CSCC to radiotherapy and chemotherapy. The effect of FTO on the malignant phenotype of CSCC was achieved through FBXW7. 

Objective To investigate the expression of ribophorin Ⅱ (RPN2) in gastric cancer (GC) and its effect on the survival of GC patients. Methods Public data of GC, RT-PCR, Western blotting and immunohistochemical (IHC) staining were employed to analysis the RNA and protein expression of RPN2 in GC. The effect of RPN2 on the clinicopathological situation and prognosis of GC patients were further investigated. Results Public chip data analysis and PCR validation in clinical samples showed that the expression levels of RPN2 mRNA in GC tissues was significantly higher than that in adjacent tissues (P < 0. 05). Western-blotting and IHC detection showed that RPN2 protein expression level was significantly up-regulated in GC tissues. Further analysis found that high RPN2 expression level in GC was significantly associated with the GC differentiation (P = 0. 005), lymph node metastasis (P = 0. 043), TNM stage (P = 0. 002), vascular invasion (P = 0. 015) and nerve invasion (P = 0. 046). The overall survival of patients with lower RPN2 expression level was significantly longer than that of patients with higher RPN2 expression level (χ 2 = 9. 296, P = 0. 002). Furthermore, univariate and multivariate analyses confirmed that high RPN2 expression level was one of the independent prognostic factors affecting overall survival in GC patients (HR = 1. 763, 95% CI: 1. 106-3. 956, P = 0. 014). Conclusion The expression of RPN2 was significantly up-regulated in GC, and high RPN2 expression level was closely related to the malignant progression and poor prognosis of GC, thus it is expected to be a biomarker for the prognosis evaluation of GC. 

Objective To detect the expression level of COX7B2 in metastatic ovarian cancer, explore its expression pattern, determine the effect of COX7B2 on the prognosis of ovarian cancer, and analyze its potential value as a diagnostic marker for ovarian cancer. Methods The RNA expression level of COX7B2 in ovarian cancer were analyzed in TCGA database. The expression of COX7B2 protein in ovarian cancer tissue samples from Handan Central Hospital was analyzed by immunohistochemical staining. Combined with the survival data, the clinical staging and prognosis were analyzed. The correlation between the expression of COX7B2 and the immune cell infiltration was analyzed. Results COX7B2 was highly expressed in ovarian cancer tissues especially in the tissues of metastatic ovarian cancer. The expression level of COX7B2 was correlated with the patient histological grade, pathological stage, lymph node metastasis, distal metastasis, and the patient survival time. The diagnosis of ovarian cancer by the expression of COX7B2 had a high accuracy. COX7B2 inhibited the infiltration of T cells, NK cells, and DC cells in the tumor microenvironment. Conclusion The COX7B2 is highly expressed in the ovarian cancer tissues, which affects the prognosis of patients. COX7B2 is expected to be served as a diagnostic marker for ovarian cancer, as well as a characteristic marker for the metastatic ovarian cancer.

Objective To investigate the effect of inhibiting the expression of TLR4 in the hypothalamus on obesity-related metabolic state. Methods After 1-week adaptive feeding, 40 four week-old male C57BL / 6 mice (SPF grade) were randomly divided into two groups, in which the mice were fed with normal diet (in NCD group) or high-fat diet (in HFD group) for 4 weeks respectively. The body weight of mice were measured once a week. The mice in each of the above groups were then randomly selected and divided into two group for the injection of TLR4 shRNA lentiviral particles or control viral particles respectively. Mice were sacrificed when reached 13-week old for further experiments. The expression of TLR4 in the hypothalamus was detected by fluorescence quantitative RT-PCR. The fasting blood glucose of the mice was detected by a blood glucose meter. The levels of serum TNF-α and IL-6 were detected by ELISA. The levels of serum low density lipoprotein-cholesterol, high density lipoprotein-cholesterol, triglyceride and total cholesterol were detected by automatic biochemical analyzer. HE staining were performed in adipose tissues. Results After TLR4-shRNA lentivirus intervention, the expression level of TLR4 in the hypothalamus was significantly decreased (P< 0. 05), and the weight gain and energy intake of high-fat fed C57BL / 6 mice were significantly decreased (P < 0. 05), the levels of inflammatory factors, triglycerides, and fasting blood glucose were significantly decreased in high-fat fed C57BL / 6 mice (P < 0. 05). The hypertrophy of adipocyte was also alleviated after TLR4-shRNA lentivirus intervention. Conclusion Down-regulation of TLR4 expression in the hypothalamus can effectively improve obesity-related inflammatory response, and metabolic disorders on glucose and blood lipids.

Objective Oral squamous cell carcinoma (OSCC) is a common cancer with high recurrence rate, high metastasis rate and poor prognosis. This study aims to further understand the role of RABL6 in the occurrence, development and prognosis of OSCC tumors. Methods The quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blotting were used to detect the expression of RABL6 in 37 frozen clinical samples and OSCC cells. The correlation between RABL6 expression and clinicopathologic features was assayed. The effects of RABL6 gene knockdown on the proliferation, apoptosis and migration of OSCC cells were studied by CCK-8, colony formation, flow cytometry, wound healing and transwell assay. Results Compared with the normal adjacent tissues, the OSCC tissues have higher expression level of RABL6, which is associated with poor prognosis. Compared with the normal oral keratinocytes group, the OSCC cells have higher expression level of RABL6. Knockdown of RABL6 inhibited the proliferation, migration and invasion of OSCC cells and induced apoptosis when compared with the si-NC group. Conclusion RABL6 acts as a tumor oncogene in OSCC, which can be a potential biomarker for predicting prognosis and a new target for OSCC therapy.

Objective To investigate the effect of total flavonoids from Selaginella uncinata (TFS) on the proliferation, migration and invasion of laryngeal squamous cell carcinoma TU177 cells and its possible mechanism. Methods TU177 cells were cultured in vitro and were divided into control group, TFS-5 μg / mL, - 15 μg / mL, - 25 μg / mL groups, miR-3662 group, miR-NC group, 25 μg / mL TFS + anti-miR-3662 group and 25 μg / mL TFS + anti-miR-NC group. CCK-8 method and colony formation assay were used to detect cell proliferation. Scratch assay was used to detect cell migration. Transwell assay was used to detect cell invasion. Western blotting was used to detect the protein expression levels of epithelial cadherin (E-Cadherin) and neural cadherin (N-Cadherin) in cells, and RT-qPCR method was used to detect the expression level of miR-3662. Results Compared with the control group, the cell inhibition rate of TFS groups was increased (P< 0. 05), but the number of colonies, the wound healing rate and the number of invasions, and the expression level of N-Cadherin protein were decreased (P < 0. 05), while the expression levels of E-Cadherin protein and miR-3662 were increased (P < 0. 05), all in a dose pendent manner. The expression level of miR-3662 in the laryngeal squamous cell carcinoma tissues was significantly lower than that in the adjacent tissues (P < 0. 05). Compared with the miR-NC group, the cell inhibition rate and the expression level of E-Cadherin protein in the miR-3662 group were increased (P< 0. 05), but the number of colonies, the wound healing rate and the number of invasions, and the expression level of N-Cadherin protein were decreased (P<0. 05). Compared with the 25 μg / mL TFS + anti-miR-NC group, the cell inhibition rate and the expression level of E-Cadherin protein in the 25 μg / mL TFS + anti-miR-3662 group were decreased (P<0. 05), but the number of colonies, the wound healing rate and the number of invasions, and the expression level of N-Cadherin protein were increased (P<0. 05). Conclusion Total flavonoids from Selaginella uncinata could inhibit the proliferation, migration and invasion of laryngeal squamous cell carcinoma TU177 cells, and its mechanism may be related to the up-regulation of miR-3662 expression in the cells.

microRNA (miRNA) is a kind of non-coding single-stranded RNA with a length of 20 ~ 24 nucleotides, which is called post-transcriptional regulatory factor. miRNA regulates its target genes after transcription. The changes of miRNA expression play important regulatory roles in the occurrence and development of many diseases. microRNA 124 (miR-124) is one of the research hotspots at present. It is lowly expressed in many types of tumors, and has played important roles in suppressing tumor growth and metastasis through regulation of cell proliferation, invasion and apoptosis. It has closely participated in the occurrence, development and prognosis of malignant tumors. This review aims to make a summary of the mechanism of miR-124 in non-malignant and malignant diseases to provide theoretical evidence for their diagnosis and treatment.

In recent years, the role of circular RNA in tumor therapy has become a research hotspot. Glycolysis is an important part of tumor metabolism. Many abnormally expressed circular RNAs are closely related to the important enzymes, cytokines and signal pathways in the process of tumor glycolysis. Binding with miRNA could inhibit the function of circular RNA, thus directly or indirectly affect the expression of downstream target proteins, promote or inhibit the process of tumor glycolysis, and also affect the resistance to chemotherapy drugs in cancer patients. This review will make a summary on circular RNA in affecting tumor glycolysis through glucose transporters and metabolic enzymes, related oncogenes and transcription factors, and cancer-related signaling pathways, and the application of circular RNA in tumor therapy and drug resistance.