Objective To explore the effects of micro RNA 502-3p (miR-502-3p) on the proliferation and apoptosis of ovarian cancer cells by targeting Casitas B-cell lymphoma ( CBL). Methods The ovarian cancer related datasets GSE66957, GSE119056 and TCGA _ OV were downloaded and the relationship between ovarian cancer and miR-502-3p or CBL was analyzed. The SKOV3 and HO8910 cell lines with overexpressed miR-502-3p or/ and CBL were constructed. The cell proliferation and apoptosis were detected by cell counting kit 8 (CCK-8), clone formation assay and flow cytometry. The effect of CBL overexpression on tumor growth was observed in tumor bearing nude mice. The targeting relationship between miR-502-3p and CBL was verified. Results Bioinformatics analysis showed that the expression level of CBL in ovarian cancer tissues was higher than that in para-cancerous tissues, while the expression level of miR-502-3p was lower in cancer tissues than in para-cancerous tissues (P < 0. 05). The expression level of CBL was related to the prognosis of ovarian cancer and the expression of cell proliferation-related genes (P < 0. 05). There was a targeting relationship between miR-502-3p and CBL. The volume and weight of tumors were significantly increased in the CBL group compared with the vector group (P< 0. 05). The expression level of CBL protein, cell viability and number of clones formed in SKOV3 and HO8910 cell lines were significantly decreased and the apoptosis rate was significantly increased in the miR-502-3p group compared with the miR-NC group (P < 0. 05). However, CBL overexpression could reverse the above changes. Conclusion miR-502-3p can inhibit the proliferation and induce the apoptosis of ovarian cancer cells by down-regulating CBL.

Objective To investigate the inhibitory effect of recombination plasminogen Kringle 5 (K5) on angiogenesis in mice. Methods Scratch assay and adhesion ability assay were used to determine the effect of recombinant K5 on the migration and adhesion of in vitro cultured mouse endothelial cells. The effect of K5 on angiogenesis was detected by utilizing in vitro angiogenesis system, wound healing and chicken embryo allantoic membrane ( CAM) angiogenesis ability assay. Results K5 (200 nmol / L) markedly inhibited the migration and adhesion of endothelial cells as well as the growth of new blood vessels. K5 also significantly inhibited wound healing in mice at the concentration of 2 to 18 mg / kg. In addition, 6, 12 and 25 mg / L K5 significantly inhibited CAM neovascularization, with the strongest inhibitory effect of K5 at 25 mg / L. Conclusion These results suggest that K5 can significantly inhibit the angiogenesis of vascular endothelial cells, which provides a theoretical basis for clinical treatment of angiogenesis

Objective To investigate the mechanism of miR-495-3p / Wnt inhibitor factor 1 (WIF1) / Wnt signaling pathway in regulating the proliferation, migration and invasion of retinoblastoma (RB) cells. Methods The relationship between differential expression of WIF1 in RB patients and the clinically adverse phenotypes was analyzed by bioinformatics. The miRNAs which bind with WIF1 were predicted and verified by dual-luciferase reporter assay. RB cells with stable overexpression of WIF1 or miR-495-3p were constructed respectively. The expressions of WIF1 protein and Wnt pathway related proteins were detected by Western blotting. In vitro assays for cell proliferation, apoptosis, migration and invasion were conducted. The volume and weight of tumors were measured in nude mice transplanted with xenograft tumors. The expression level of WIF1 in RB tissues was analyzed by immunohistochemistry. Results WIF1 was lowly expressed in RB tissues, and the low expression of WIF1 was related to migration and invasion. WIF1 was the target of miR495-3p, which was verified by dual-luciferase reporter assay. Overexpression of WIF1 could inhibit cell proliferation, migration and invasion, affect cell cycle, and induce apoptosis. In vivo assay showed that the overexpression of WIF1 in RB tissues could inhibit tumor growth. The expression levels of β-catenin and c-Myc proteins were downregulated after WIF1 was overexpressed (P< 0. 05). Overexpression of miR-495-3p could reduce the expression level of WIF1 protein, increase the expression levels of β-catenin and c-My, promote cell proliferation, migration and invasion, and inhibit apoptosis (P< 0. 05). The increased expression of WIF1 could reverse the effect of miR-495- 3p on proliferation, migration, invasion and apoptosis of RB cells (P< 0. 05). Conclusion miR495-3p could inhibit proliferation, migration and invasion of RB cells, and induce apoptosis by WIF1 / Wnt signaling pathway.

Objective To investigate the effect of miR-142-5p on epithelial-mesenchymal transition (EMT) and chemosensitivity of gemcitabine (GEM) in human cholangiocarcinoma cell line QBC939 by targeting myeloid cell leukemin-1 ( MCL-1 ). Methods Human cholangiocarcinoma cell line QBC939 and human normal bile duct epithelial cell line HIBEC were cultured in vitro and then randomly divided into control group, miR-142-5p negative control (NC) group and miR-142- 5p mimics group. The mRNA expression levels of miR-142-5p and MCL-1 in QBC939 cells and HIBEC cells were detected by real-time quantitative PCR (RT-qPCR); the migration and invasion of QBC939 cells were determined by wound healing assay and transwell assay; Western blotting was used to detect the expression level of E-cadherin, N-cadherin and Vimentin in QBC939 cells; dualluciferase assay was used to verify the relationship between miR-142-5p and MCL-1; QBC939 cells transfected with miR-142-5p were treated with GEM at a concentration of 0. 625 μg / mL. The viability and apoptosis of QBC939 cells were measured by MTT assay and Annexin V-FITC / PI kit respectively. Results The expression level of miR-142-5p was lower and the expression level of MCL-1 was higher in cholangiocarcinoma QBC939 cells than in human normal bile duct HIBEC cells. According to the prediction of TargetScan Human Database, there was a binding site for miR-142-5p on the 3′UTR region of MCL-1 mRNA. Compared with MCL-1-3′UTR-WT + miR-142-5p NC group, the luciferase activity of MCL-1-3′UTR-WT + miR-142-5p mimics group was significantly decreased (P< 0. 05). The expression level of miR-142-5p and E-cadherin were significantly increased (P< 0. 05), the expression level of MCL-1, N-cadherin and vimentin were significantly decreased (P< 0. 05), and the number of migratory and invasive cells were profoundly reduced in miR-142-5p mimics group compared with control group and miR-142-5p NC group (P < 0. 05 for all). The survival rate of QBC939 cells in miR-142-5p mimics + GEM group was significantly lower and the apoptosis rate was higher than that in miR-142-5p NC + GEM group (P < 0. 05). Conclusion Overexpression of miR-142-5p can inhibit the EMT of and increase the chemosensitivity of GEM to cholangiocarcinoma cells by targeting MCL-1.

Objective To investigate the effect of miR-130a-3p on glucose metabolism, proliferation and migration of liver cancer cells by targeting PDK1, a key enzyme of glucose metabolism, so as to provide a new target for the diagnosis and treatment of liver cancer. Methods The expression of miR-130a-3p in clinical tissue samples and cell lines of hepatocellular carcinoma was detected by qRT-PCR. Cell proliferation was measured by clone formation and CCK-8. Cell migration was measured by transwell and wound healing assay. Western blotting was used to detect the expression of related proteins. The production of lactic acid and ATP in cells was assayed by lactate assay kit and ATP assay kit respectively, and the PDH activity in cells was assayed by PDH activity assay kit. The dual-luciferase reporter gene assay was used to verified the binding of miR-130a-3p to PDK1 3′ UTR. Results The expression level of miR-130a-3p was lower in liver cancer tissues and cells, and it was related to the size of tumor tissues and TNM stage. Overexpression of miR-130a-3p could enhance the activity of PDH in hepatoma cells, reduce the production of lactic acid and ATP, and inhibit the proliferation and migration of hepatoma cells. miR-130a-3p could bind to PDK1 and regulate its expression. Inhibition of PDK1 expression could reverse the enhancing effects of miR-130a-3p overexpression on cell proliferation and migration, and the effects on PDH activity, production of lactic acid and ATP were also reversed. Conclusion miR-130a-3p affects glycolysis and malignant progression of hepatoma cells by targeting PDK1.

Objective To investigate the effects of Guilingji on the expression of Fas/ FasL and neuronal apoptosis in cortex and hippocampus of mice with Alzheimer’s Disease (AD). Methods AD mouse model was constructed and randomly divided into the model group, donepezil group and low-, medium-and high-dose Guilingji groups. Morris water maze test was used to test the learning and memory ability of mice before and after intervention. HE staining was used to determine the pathological changes of cortical and hippocampal neurons. TUNEL staining was used to measure neuronal apoptosis. Western blotting and real-time PCR were used to detect protein and mRNA expression levels of Fas and FasL. Results The learning and memory ability of mice was significantly improved (P< 0. 05), the pathological damage and apoptosis of neurons in cortex and hippocampus greatly alleviated, and the protein and mRNA expression levels of Fas and FasL markedly decreased in Gulingji groups and donepezil group compared with model group (P < 0. 05 for all). Conclusion Gulingji may inhibit the apoptosis of cortical and hippocampal neurons and improve the learning and memory ability of AD model mice by inhibiting the expression of Fas/ FasL. 

Objective To explore the effects of sufentanil on autophagy, apoptosis and proliferation of cervical cancer cells SiHa. Methods SiHa cells were treated with sufentanil (3. 125, 6. 25, 12. 5, 25, 50, 100, 200, 400, 800 nmol / L ) . Cell viability was measured by CCK8. Four concentrations of sufentanil (0, 25, 50, 100 nmol / L) were then selected to treat SiHa cells for 24 h for the following experiments. The apoptosis and proliferation of SiHa cells were measured by flow cytometry and colony formation assay. The level of LC3 in each group was measured by immunofluorescence method. The levels of autophagy-related proteins (LC3Ⅱ/ LC3Ⅰ, Beclin1, ATG7 ), proliferation-and apoptosis-related proteins ( P21, Survivin ) were detected by Western blotting. The expression levels of cell proliferation-related genes (Ki67, PCNA) were detected by RT-PCR. Results The viability of SiHa cells decreased with the increased concentration of sufentanil and the effect was in a dose-dependent manner. The expression levels of LC3Ⅱ/ LC3 Ⅰ, Beclin1 and ATG7 in SiHa cells treated with 50 nmol / L and 100 nmol / L sufentanil were signif- icantly higher than those treated with 0 nmol / L sufentanil. The immunofluorescence method showed that the fluorescence signal of LC3 in SiHa cells treated with 50 and 100 nmol / L sufentanil was higher than that in cells treated with 0 nmol / L sufentanil. Flow cytometry showed that the apoptosis rate of SiHa cells treated with 50 and 100 nmol / L sufentanil was higher than that of cells treated with 0 nmol / L sufentanil. Clone formation assay showed that the clonal formation rate of SiHa cells was decreased after 50 and 100 nmol / L sufentanil treatment. The expression levels of Survivin, Ki67 and PCNA were decreased, and the expression level of P21 was increased. Conclusion Sufentanil can promote autophagy and apoptosis of cervical cancer cells, and inhibit their proliferation, which may be through the regulation of autophagy-, apoptosis- and proliferation-related proteins or genes.

Objective To investigate the effect of microRNA-101-3p (miR-101-3p) on alveolar epithelial cell apoptosis induced by Streptococcus pneumoniae (SP) through targeting tumor necrosis factor receptor related factor 6 (TRAF6). Methods A549 cells were cultured. Dual-luciferase reporter assay was used to detect the binding of miR-101-3p with TRAF6. Real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to detect the mRNA expression levels of miR-101-3p and TRAF6. Western blotting was used to detect the protein expression levels of TRAF6, activated caspase-3 (C-caspase-3) and C-caspase-9. Flow cytometry assay was used to detect cell apoptosis rate. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of interleukin-6 (IL-6) and IL-10 in the cell supernatant. Results TRAF6 was targeted and negatively regulated by miR-101-3p (P < 0. 05). After SP induction, the expression level of miR-101-3p was decreased and the expression level of TRAF6 was increased, the level of IL-6 was increased and the level of IL-10 was decreased, the apoptosis rate was increased, the expression levels of C-caspase-3 and C-caspase-9 was increased (P< 0. 05). Overexpression of miR-101-3p could attenuate the injury induced by SP in A549 cells. Overexpression of TRAF6 could partially reverse the protective effect of miR-101-3p on SP-induced apoptosis in A549 cells (P< 0. 05). Conclusion miR-101-3p negatively regulates TRAF6 to protect alveolar epithelial cells from SP-induced apoptosis and inflammatory damage. 

Objective To investigate the expression level of long non-coding RNA HOX transcript antisense RNA ( LncRNA HOTAIR) in the serum of patients with diabetic nephropathy (DN), and to explore its value in the diagnosis of DN. Methods From January 2020 to January 2021, 80 patients with type 2 diabetic nephropathy (DN group) and 80 patients with type 2 diabetes without kidney damage (T2DM group) who were hospitalized in the Endocrinology and Nephrology department of Huai’an Hospital Affiliated to Xuzhou Medical University were selected as the study subjects, and 75 healthy non-diabetic patients who visited our hospital’s physical examination center during the same period were selected as the control group. Real-time fluorescent quantitative PCR (qRT-PCR) method was used to detect the expression level of LncRNA HOTAIR in the serum of subjects. ROC curve was used to evaluate the clinical diagnostic value of serum LncRNA HOTAIR for DN. Results The expression level of LncRNA HOTAIR in T2DM and DN patients was significantly higher than that in the healthy control group (P < 0. 05), and the expression level of LncRNA HOTAIR in the DN group was significantly higher than that in the T2DM group. The expression level of LncRNA HOTAIR in the serum of DN patients increased significantly with the increase of the urine protein content (P< 0. 05). It was positively correlated with the urine protein content,fasting blood glucose, glycosylated hemoglobin, triglycerides, and low-density lipoprotein ( P < 0. 05), and negatively correlated with glomerular filtration rate ( P < 0. 05). The area under the curve (AUC) of serum LncRNA HOTAIR for diagnosis of DN was 0. 885 (95 % CI: 0. 824- 0. 946), the sensitivity was 92. 5 % , and the specificity was 86. 3 % . Conclusion The expression level of serum LncRNA HOTAIR is related to the progress of DN in T2DM patients, and is expected to be a biomarker for predicting the occurrence of DN in T2DM patients 

Objective To explore the effect of eucommia water extract on the apoptosis of neurons in rats with cerebral infarction. Methods A total of 40 rats were randomly divided into control group, model group, eucommia water extract group and Ravoxertinib group, with 10 rats in each group. The cerebral cortex damage was determined by HE staining. The apoptosis of cerebral cortex cells was assayed by TUNEL staining. The levels of oxidative stress indexes [ superoxide dismutase (SOD), malondialdehyde (MDA), cysteine aspastic protease 3 (Caspase-3)] were measured by ELISA. The extracellular signal regulated kinase 1 / 2 (ERK1 / 2) signaling in cerebral cortex was detected by Western blotting. Results HE staining showed no obvious pathological changes of brain tissues in control group, but significant pathological damage of brain tissues in model group (P < 0. 05). Eucommia water extract group and Ravoxertinib could significantly relieve the damage of brain tissues (P < 0. 05). The apoptosis rate of neurons, the levels of Caspase-3 and MDA, and the expression levels of p-ERK1 / 2 / ERK1 / 2 and Bax were significantly increased and the level of SOD was significantly decreased in model group compared with the control group (P< 0. 05). These indexes were significantly reversed in eucommia water extract group and Ravoxertinib group as compared with the model group (P< 0. 05). There was no significant difference in the indexes between the eucommia water extract group and Ravoxertinib group (P > 0. 05). Conclusion Eucommia water extract can regulate biological behaviors of neurons and oxidative stress response in brain tissues by regulating ERK1 / 2 signaling pathways, thereby playing roles in the treatment of cerebral infarction in rats. 

Objective To investigate the effect of autologous tooth-bone graft combined with platelet-rich fibrin on peri-implant defects and the underlying mechanism. Methods A total of 40 New Zealand white rabbits were randomly divided into the experimental groups A, B, C and the blank control group. Under the condition of general anesthesia, a minimally invasive surgical procedure was performed to extract the animals’maxillary anterior teeth. The tooth socket of rabbits in the experimental group A were implanted with autologous tooth-bone grafts combined with platelet-rich fibrin. That in experimental group B was only implanted with autogenous tooth-bone grafts. Rabbits in experimental group C experienced only the implantation with platelet-rich fibrin, and no material was implanted in blank control group. The activity of alkaline phosphatase (ALP), the gray values of typeⅠcollagen, osteoprotegerin (OPG) and its ligand (RANKL), the mRNA expression levels of Runx2, ALP and platelet reactive protein-1 were compared and analyzed. Results The ALP activity, the gray values of typeⅠcollagen and OPG, the mRNA expression levels of Runx2, ALP and platelet reactive protein-1 in the experimental groups were higher than those in the blank control group. The values of the above indexes in the experimental group A were significantly higher than those in experimental groups B and C ( P < 0. 05). Although the gray values of RANKL in each group were significantly increased with time (P< 0. 05), there was no significant difference in the gray values of RANKL between the experimental groups and the blank control group at each time point (P > 0. 05). Conclusion The combination of platelet-rich fibrin enhanced the repair of peri-implant defects in the process of autologous tooth-bone transplantation.

Objective To explore the expression of inducible T cell costimulatory molecules in colon cancer tissues and their correlation with long-term survival of colon cancer patients. Methods A total of 206 colon cancer patients who underwent surgical resection of colon tissue in Xinhua Hospital Affiliated to Dalian University from May 2018 to May 2019 were selected. Their colon cancer tissues and paracancerous tissues were used for the detection of inducible T-cell co-stimulator (ICOS) expression by fluorescence quantitative PCR. The relationships between the expression level of ICOS and the clinicopathological characteristics or the long-term survival were analyzed. Results The expression level of ICOS was significantly lower in colon cancer tissues (8. 73 ± 2. 25) than that in adjacent tissues (18. 54 ± 3. 26) (P< 0. 05). The expression level of ICOS in colon cancer tissues was related to tumor diameter, distant metastasis, clinical stage, lymph node metastasis (P< 0. 05), and no relation was found between ICOS expression level and age, gender, tumor location, and degree of differentiation (P > 0. 05). The expression level of ICOS in 206 colon cancer tissues was low in 117 cases, and high in 89 cases. Logistic regression analysis showed that ICOS expression was related to tumor size, clinical stage, lymph node metastasis, and distant metastasis (P< 0. 05), but not related to age, gender, lymph node metastasis, tumor location, and degree of differentiation (P > 0. 05). The 206 colon cancer patients were followed up until September 30, 2021, the survival rate was 83. 01 % (171 / 206), the mortality rate was 16. 99 % (35 / 206), and the median survival time was (23. 5 ± 3. 3) months. The median survival time of the 89 colon cancer patients with high expression level of ICOS was (27. 4 ± 3. 2) months, with a 95 % CI of 2. 24-6. 147. The median survival time of the 117 colon cancer patients with low expression level of ICOS was (16. 3 ± 4. 3) months, and the 95 % CI was 1. 458-5. 237. The median survival time of colon cancer patients with high expression level of ICOS was significantly higher than that of patients with low expression level of ICOS (P<0. 05). Conclusion ICOS is lowly expressed in colon cancer tissues, and its expression level is positively correlated with the long-term survival of patients. The expression level of ICOS can be used as an important evaluation index for the prognosis and survival of colon cancer patients.

N⁶ -methyladenosine ( m6A) modification is a post-transcriptional change of RNA molecule. m6A was first discovered in cell mRNA in 1970s, but the extensive investigation on m6A modification did not begin until the genome-wide m6A localization method was developed. The role of m6A in regulating mRNA and its functional importance in various cell types have been verified in many studies, and the mechanism of m6A in regulating gene expression in a variety of physiological processes has been increasingly studied recently. In this review, we summarized recent advances in m 6A modification in immune response, and discussed the regulatory role of m6A modification in innate and adaptive immune responses, and immune system development. 

Psoriasis is a common chronic and recurrent inflammatory skin disease. The disorder of keratinocyte (KC) proliferation and differentiation is one of its pathogeneses, but the underlying molecular mechanisms are still largely unknown. The extracellular signal regulated kinase (ERK) signaling pathway is reported to play an important role in KC proliferation and differentiation. MicroRNA (miRNA), long non-coding RNA (lncRNA), cytokines and other upstream molecules of ERK signaling pathway are also involved in the process. The purpose of this article is to review the mechanism of ERK signaling pathway in hyperproliferation of KCs in psoriasis

Endothelial cells are important components of the heart, blood vessels and lymph vessels. They play a crucial role in regulating the physiological processes of the cardiovascular system. Endothelial-mesenchymal transition (EndMT) is the transition of endothelial cells to mesenchymal cells, and it has been reported to be involved in atherosclerosis. Various cytokines are changed during the process of EndMT. The study of EndMT may provide insight for the mechanism and treatment of atherosclerosis. 

Neurodegenerative diseases are caused by acute and chronic pathological factors, and are characterized by loss and degeneration of nerve cells. It is the second most common cause of death. The prevention of neurodegenerative diseases has become an emerging field of public health, which poses great challenges to our society. Melatonin is a kind of pineal hormone, which has many physiological functions, including regulating circadian rhythm, scavenging free radicals, inhibiting biomolecular oxidation and neuroinflammation. It has been reported that patients with neurodegenerative diseases have lower melatonin levels. Melatonin plays the role of neuroprotection through some pathophysiological mechanisms and signaling pathways, and it is implied to be a new treatment for neurodegenerative diseases. This paper reviews the role and research progress of melatonin in neurodegenerative diseases such as Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, amyotrophic lateral sclerosis, vascular dementia and multiple sclerosis.