Objective To explore the effect and molecular mechanism of proanthocyanidins on the improvement of osteoporosis based on Wnt signaling pathway. Methods The osteoporosis rat model was constructed in 50 SD female rats by ovariectomy, then they were randomly divided into model group, alendronate sodium group, proanthocyanidins low-, medium-, and high- dose groups. Another 10 SD rats were recorded as the control group, and the bilateral ovaries of rats in the control group were turned over only without resection. Rats in the alendronate sodium group were given 1 mg / kg alendronate sodium by gavage. Rats in the procyanidins low-, medium-, and high dose groups were given procyanidins by gavage at the dose of 15, 25 and 35 mg / kg respectively. Rats in the control group and the model group were given the same amount of normal saline injection by gavage, once a day for 12 weeks. The levels of serum oxidative stress indexes [malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px)] and bone metabolism indexes [tartrate resistant acid phosphatase (TRAP), type I procollagen carboxyl-terminal peptide (PICP), bone Gla-protein (BGP)] in rats of each group were measured by enzyme-linked immunosorbent assay ( ELISA). Bone density scanner was used to determine the value of femoral bone mineral density (BMD). Hematoxylin-eosin (HE) staining was used to observe the morphological changes of bone tissues in rats. Real time fluorescence quantitative polymerase chain reaction (RT-qPCR) and Western blotting (WB) were used to detect the mRNA and protein expression levels of Wnt, β-catenin, and osteogenic marker proteins [runt related transcription factor 2 (Runx2), Osterix]. Results Compared with the control group, the serum MDA and TRAP levels in the model group, the alendronate sodium group and the procyanidins low-, medium- , and high-dose groups were increased, the levels of SOD, GSH-Px, PICP and BGP were decreased (P< 0. 05). Compared with the model group, the serum MDA and TRAP levels in the alendronate sodium group and the procyanidins low-, medium-, and high-dose groups were decreased, the levels of SOD, GSH-Px, PICP and BGP were increased (P < 0. 05). In the control group, the bone trabeculae were arranged reticulated and formed a dense network. In the model group, the number of trabeculae was decreased, the arrangement was sparse and disordered, and the continuity was poor. Compared with the model group, the number and structure of the bone trabeculae in the alendronate group and the procyanidins low-, medium-, and high-dose groups were improved to varying degrees. Compared with the control group, the mRNA and protein expression levels of Wnt, β-catenin, Runx2, Osterix bone tissues of the model group, the alendronate sodium group and the procyanidins low-, medium-, and high-dose groups were decreased (P < 0. 05). Compared with the model group, the mRNA and protein expression levels of Wnt, β-catenin, Runx2, Osterix in the alendronate sodium group and the procyanidins low-, medium-, and high-dose groups were increased (P < 0. 05). Conclusion Procyanidins can effectively improve the oxidative stress and bone metabolism in osteoporosis rats, and play a role in the treatment of osteoporosis. It is speculated that this effect is achieved by the inhibition of the peroxidative stress response and the promotion of the expressions of Wnt, β-catenin and osteogenic marker proteins Runx2 and Osterix through Wnt pathway.

Objective To investigate the role of CXC motif chemokine ligand 8 (CXCL8) in oral squamous cell carcinoma (OSCC). Methods The expression level of CXCL8 in OSCC tissues and cells was detected by RT-qPCR and Western blotting, the correlation between CXCL8 expression level and the clinicopathological features of OSCC patients was analyzed. The effect of CXCL8 knockdown on cell proliferation, apoptosis, migration, invasion and epithelial-mesenchymal transition (EMT) was detected by cell counting kit 8 (CCK-8) method, colony formation assay, flow cytometry, wound healing assay, transwell assay and Western blotting. Results CXCL8 was significantly up-regulated in OSCC tissues and cells. The high expression level of CXCL8 was significantly correlated with tumor size, TNM staging and distant metastasis. CXCL8 knockdown can inhibit the proliferation, migration, invasion and EMT of SCC9 cells, and promote the apoptosis. Conclusion CXCL8 is up-regulated in OSCC, and the suppression of its expression can inhibit the proliferation and metastasis of SCC9 cells.

Objective To investigate the effect of hypoxia-inducible factor 1α (HIF-1α) gene interference on high glucose (HG) -induced apoptosis, inflammation and extracellular matrix deposition in renal tubular epithelial cells (HK-2). Methods HK-2 cells were divided into 4 groups: control group, HG group, HG + shRNA-NC group, HG + HIF-1α-shRNA group. The expression levels of HIF-1α and neutrophil gelatinase-associated lipocalin (NGAL) were detected by quantitative real-time polymerase chain reaction. The cell proliferation, apoptosis and the levels of inflammatory factors were assayed by cell counting kit-8, flow cytometry and enzyme-linked immunosorbent assay, respectively. The protein expression levels were detected by Western blotting. Results Compared with the HG group, the mRNA and protein expression levels of HIF-1α and NGAL were decreased in the HG + HIF-1α-shRNA group, the cell proliferation rate was increased, the apoptosis rate was decreased, the levels of interleukin-6 (IL-6) and tumor necrosis factor-α were decreased, the level of IL-10 was increased, the expression levels of tissue inhibitors of metalloproteinase l and collagen type I protein were decreased, and the expression level of matrix metalloproteinase 9 protein was increased in the HG + HIF-1α-shRNA group (P< 0. 01). Conclusion HIF-1α gene interference alleviates HG-induced apoptosis, inflammation and extracellular matrix deposition in HK-2 cells.

Objective To investigate the expression of PTEN-Long in gastric cancer cells and its effect on the proliferation and apoptosis of gastric cancer cells (SGC-7901). Methods RT-qPCR and Western blotting were performed to detect the expression levels of PTEN-Long mRNA and protein in gastric cancer cells, and the cell line for further studies was selected. The recombinant lentiviral vector carrying the PTEN-Long gene was transfected into the SGC-7901 cells (the PTEN-Long group), cells in the NC group were transfected with the empty lentiviral vector, and cells in the control group were not transfected, and the transfection efficiency was detected. Cell proliferation was assayed by MTT method and Edu staining. Cell cycle and apoptosis were measured by flow cytometry. The expression levels of P27, cyclin D1 and PI3K/ AKT pathway related proteins were detected by Western blotting. Results The expression levels of PTEN-Long mRNA and protein in gastric cancer cells were greatly reduced (P < 0. 05). SGC-7901 cell line was selected for the subsequent experiments due to its lowest expression level of PTEN-Long among the detected gastric cancer cell lines. RT-qPCR and Western blotting confirmed that the lentivirus transfection was successful. Compared with the control group and the NC group, the cell proliferation in the PTEN-Long group was greatly reduced, the proportion of cells in the G0 / G1 phase was greatly increased, and the proportions of cells in the G2 / M and S phases were greatly decreased (P< 0. 05), the apoptosis rate was greatly increased. The expression level of P27 protein was greatly increased, and the protein expression levels of p-PI3K, p-AKT and cyclin D1 were greatly decreased in the PTEN-Long group (P < 0. 05). Conclusion PTEN-Long is down-regulated in gastric cancer cells, and the overexpression of PTEN-Long can inhibit the proliferation of gastric cancer cells and promote cell apoptosis by inhibiting the activation of PI3K/ AKT signaling pathway. 

Objective To explore the potential role of programmed cell death ligand 1 ( PD-L1) in icotinib resistance by using epidermal growth factor receptor (EGFR) -mutant non-small cell lung cancer (NSCLC) cells. Methods The expression levels of p-EGFR and PD-L1 in A549, HCC827 and PC-9 cells were detected by real-time fluorescent quantitative PCR ( qRT-PCR) and Western blotting. MTT method was used to evaluate the sensitivity of HCC827 and PC-9 cells to icotinib. PD-L1 knockout cells were used to evaluate the effect of PD-L1 on the sensitivity of EGFR-mutant NSCLC cells to icotinib, and the patient-derived tumor xenografts mouse model was used for the in vivo experiments. Results Compared with the A549 cells, the expression levels of p-EGFR and PD-L1 were higher in the HCC827 and PC-9 cells. Icotinib significantly inhibited the proliferation and enhanced the apoptosis of HCC827 and PC-9 cells. Knockout of PD-L1 significantly reduced the inhibition of icotinib on HCC827 and PC-9 cells. Compared with the sh-NC + icotinib group, PD-L1 knockout reduced the sensitivity of EGFR-mutant NSCLC cells to icotinib in vivo. Conclusion Icotinib could inhibit EGFR-mutant NSCLC cells, and PD-L1 contribute to the icotinib sensitivity in EGFR-mutant NSCLC cells. 

Objective To analyze the relationship between miR-3682-3p and PI3K/ AKT signaling pathway in the development of hepatocellular carcinoma, in order to provide new ideas for the treatment of hepatocellular carcinoma. Methods The expression levels of miR-3682-3p and PI3K/ AKT in MIHA human normal hepatocytes and SMMC-7721 human hepatoma cells were detected by qRT-PCR. SMMC-7721 human hepatoma cells were divided into miR-3682-3p mimic group, miR-3682-3p mimic NC group, miR-3682-3p inhibitor group and miR-3682-3p inhibitor NC group by transfection of different plasmids. The interaction between miR-3682-3p and PI3K was analyzed by dual-luciferase gene reporter assay. Western blotting, qRT-PCR, flow cytometry, wound healing assay and transwell assay were used to analyze the proliferation, invasion and apoptosis of cells in different groups. Results miR-3682-3p could targetly regulate and activate the PI3K/ AKT signaling pathway, and enhance the proliferation, migration and invasion of hepatoma cells, reduce the apoptosis. When the expression of miR-3682-3p was inhibited, the PI3K/ AKT pathway was suppressed, the proliferation, migration and invasion of hepatoma cells were decresed, and the apoptosis was increased. Conclusion PI3K is a target of miR-3682-3p, and miR-3682-3p promotes the in vitro metastatic activity of SMMC-7721 human hepatoma cells by activating the PI3K/ AKT pathway.

Objective To investigate the effect of PCA3 on the proliferation and apoptosis of Hodgkin’s lymphoma cell line L428 and its possible mechanism. Methods Lymphoma tissues and normal adjacent tissues from 39 patients with Hodgkin’s lymphoma were collected. The expression levels of PCA3 and miR-185 in the tissues were dectected by RT-qPCR. L428 cells were divided into si-NC group, si-PCA3 group, si-PCA3 + NC group, si-PCA3 + miR-185 inhibitor group. CCK-8 was used to detect cell proliferation. Flow cytometry was used to detect cell cycle and apoptosis. Western blotting was used to detect the protein expression levels of pro-caspase 3 and cleaved-caspase 3. RT-qPCR was used to detect the expression levels of PCA3 and miR-185. The dual luciferase gene reporter assay was used to verify the regulatory relationship between PCA3 and miR-185. Results The expression level of PCA3 was increased and the expression level of miR-185 was decreased in the lymphoma tissues (P< 0. 05). Transfection of si-PCA3 could inhibit cell proliferation, arrest cell cycle, and promote cell apoptosis in L428 cells. PCA3 targets to the miR-185 and negatively regulate its expression. Co-transfection of si-PCA3 and miR-185 inhibitor could reduce the effect of transfection of si-PCA3 on cell proliferation, cell cycle and apoptosis. Conclusion Interfering the expression of PCA3 could target and negatively regulate miR-185 to inhibit the proliferation of lymphoma cells L428 and promote the apoptosis of L428 cells.

Objective To investigate the effect of bergapten (BG) on the inhibition of high glucose (HG) induced inflammatory factors expression and apoptosis in renal tubular epithelial cells by down-regulation of miR-503. Methods The renal tubular epithelial cells were divided into control (Con) group, high glucose ( HG) group, high glucose + BG low-, medium- and high concentration (HG + BG-L, -M, -H) group, HG + inhibitor-NC group, HG + miR-503 inhibitor group, HG + BG + miR-NC group, HG + BG + miR-503 group. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of inflammatory factors tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), flow cytometry was used to monitor the cell apoptosis, and Western blotting was used to determine the expression levels of apoptotic proteins cleaved-caspase 3 and cleaved-caspase 9, and the expression level of miR-503 was determined by qRT-PCR. Results The apoptosis rate, the levels of TNF-α, IL-6, and the expression levels of cleaved-caspase 3, cleaved-caspase 9 protein and miR-503 in renal tubular epithelial cells in the HG group were all higher than those in the Con group (all P< 0. 05). The apoptosis rate, the levels of TNF-α, IL-6, and the expression levels of cleaved-caspase 3, cleaved-caspase 9 protein, miR-503 of renal tubular epithelial cells in the HG + BG-L group, the HG + BG-M group, and the HG + BG-H group were all lower than those in the HG group (all P < 0. 05), and the inhibitory effects of BG were concentration dependent. After interfering with miR-503, the expression levels of miR-503, cleaved-caspase 3 and cleaved-caspase 9, the levels of TNF-α, IL-6, and the apoptosis rate in renal tubular epithelial cells in the HG + miR-503 inhibitor group were lower than those in the HG + inhibitor-NC group (all P < 0. 05). The expression levels of miR-503, cleaved-caspase 3 and cleaved-caspase 9, the levels of TNF-α, IL-6, and the apoptosis rate in renal tubular epithelial cells in the HG + BG + miR-503 group were higher than those in the HG + BG + miR-NC group (all P< 0. 05). Conclusion BG inhibits the high glucose induced expression of inflammatory factors and apoptosis of renal tubular epithelial cells in a concentration-dependent manner by down-regulation of the miR-503 expression.

Objective To investigate the effect of luteolin on lipopolysaccharide ( LPS) -induced intestinal epithelial cell pyroptosis via NOD-like receptor family pyrin domain containing 3 (NLRP3) / caspase-1 pathway. Methods Human intestinal epithelial cells HIEC-6 were cultured in vitro, and induced by LPS (1. 0 μg / mL), then the cells were divided into control group, LPS group, LPS + (25, 50, 100 μmol / L) luteolin groups, luteolin + NLRP3 activator nigericin sodium (NSS) group. LPS (1. 0 μg / mL) was added in each group except for the control group. Cell viability was assayed by the MTT method. Monolayer epithelial transmembrane resistance (TEER) was determined by the resistance meter. The levels of lactate dehydrogenase (LDH), tumor necro- sis factor-α ( TNF-α) and interleukin-6 ( IL-6) in each group were measured by the enzyme-linked immunosorbent (ELISA) method. The expression levels of pyroptosis-related proteins (GS-DMD, GSDMD-N) and NLRP3, caspase-1, IL-1β in each group were detected by Western blotting (WB). Results Compared with the control group, the cell viability and TEER value in the LPS group were significantly reduced, and the levels of cell supernatant LDH, TNF-α, IL-6, and the expression levels of GSDMD, GSDMD-N, NLRP3, caspase-1 and IL-1β were significantly increased (P< 0. 05). Compared with the LPS group, the cell viability and TEER value in the LPS + (25, 50, 100 μmol / L) luteolin groups were significantly higher, and the levels of cell supernatant LDH, TNF-α, IL-6, and the expression levels of GSDMD, GSDMD-N, NLRP3, caspase-1, IL-1β were significantly reduced (P< 0. 05). Compared with the LPS + 100μmol / L luteolin group, the cell viability and TEER value in the luteolin + NSS group were significantly reduced, and the levels of cell supernatant LDH, TNF-α, IL-6, the expression levels of GSDMD, GSDMD-N, NLRP3, caspase-1, IL-1β were significantly increased (P< 0. 05). Conclusion Luteolin can alleviate the pyroptosis and inflammatory injury of human intestinal epithelial cells HIEC-6 induced by LPS, this may be related to the inhibition of NLRP3 / caspase-1 signaling pathway.

Objective To investigate the effect of cinnamaldehyde on apoptosis of nasal mucosa cells in rats with allergic rhinitis (AR) and its mechanism. Methods The AR model was established in rats and the rats were randomly divided into AR group, low-, medium-and high-dose cinnamaldehyde groups and loratadine group, another 10 rats were selected as the sham operation group (sham group). The apoptosis of nasal mucosa cells, the level of serum IL-6, and the expression levels of IL-6, JAK2, STAT3 mRNAs and JAK2, p-JAK2, STAT3, p-STAT3, Bcl-2, Bax proteins were detected. Results Compared with the sham group, the level of serum IL-6, the expression levels of IL-6 mRNA and P-JAK2 / JAK2, P-STAT3 / STAT3 and Bcl-2 proteins of nasal mucosa cells were increased in the AR group (P < 0. 05), while the apoptosis index (AI) and Bax protein expression level were decreased (P< 0. 05). Compared with the AR group, the level of serum IL-6, the expression levels of IL-6 mRNA and P-JAK2 / JAK2, P-STAT3 / STAT3 and Bcl-2 proteins in rats nasal mucosa cells in the low-dose, medium-dose and high-dose cinnamaldehyde groups and the loratadine group were decreased (P< 0. 05), while the AI and Bax expression level were increased (P< 0. 05). Conclusion Cinnamaldehyde can promote the apoptosis of nasal mucosal cells in AR rats, and the mechanism may be related to the inhibition of IL-6 / JAK2 / STAT3 signaling pathway. 

Objective To explore the possible mechanism of rhamnetin on protecting brain damage in rats with diabetes mellitus (DM) undergoing cerebral ischemia-reperfusion (CI/ R). Methods The model of DM rats was constructed, and CI/ R was prepared by suture-occluded method. They were administered intragastrically with rhamnetin (50, 100, 200 mg / kg). The brain water content, brain index, cell apoptosis, serum inflammatory factors, markers for cell apoptosis and inflammatory response, expression levels of Notch1 and P38 mitogen-activated protein kinase (P38 MAPK) proteins were detected 14 days after administration. Notch1 activator Jagged1 was applied to observe the changes of expression levels of Notch1 and P38 MAPK proteins. Results After intervention with medium- and high- doses of rhamnetin, the number of error times in step-down test, the brain index and brain water content, the apoptosis rate, the levels of IL-6, IL-1β, TNF-α, and the expression levels of cleaved caspase-3 / caspase-3, cleaved caspase-9 / caspase-9, p-Notch1 / Notch1 and p-P38 MAPK/ P38 MAPK were decreased, while the number of times that the rat entering extraneous arms and the level of IL-10 were increased (P< 0. 05). Rhamnetin could inhibit the Jagged1 induced activation of Notch1-p38 MAPK signaling (P<0. 05). Conclusion Rhamnetin can protect DM rats undergoing CI/ R from brain tissue damage, its mechanism may be related to the alleviation of inflammatory response through down-regulation of Notch1-p38 MAPK signaling.

Objective To explore the effect of total flavonoids from Valeriana jatamansi Jones (TFV) on gastric cancer cells AGS proliferation, migration and invasion and its possible mechanism. Methods In vitro cultured human gastric cancer cells AGS were randomly divided into Con group, TFV-L group, TFV-M group, TFV-H group, miR-NC group, miR-4429 group, TFV + anti-miR-NC group, and TFV + anti-miR-4429 group. The expression of miR-4429, cell proliferation, clone formation ability, cell migration and invasion were detected by qRT-PCR, CCK-8, plate colony formation assay, wound healing assay and transwell assay, respectively. The expression levels of E-cadherin and N-cadherin proteins were detected by Western blotting. Results Compared with the Con group, the expression levels of E-cadherin, proliferation inhibition rate and miR-4429 expression level in the TFV-L, TFV-M, and TFV-H groups were increased (P< 0. 05), while the expression level of N-cadherin and the wound healing rate were decreased (P< 0. 05), the number of colonies formed and the number of invasive cells were decreased (P < 0. 05), all in a dose-dependent manner. Compared with the miR-NC group, the cell proliferation inhibition rate and the expression level of E-cadherin in the miR-4429 group were increased (P< 0. 05), while the number of cell clones and the number of invasive cells were decreased (P< 0. 05), the wound healing rate and the expression level of N-cadherin were decreased (P < 0. 05). Compared with the TFV + anti-miR-NC group, the cell proliferation inhibition rate and the expression level of E-cadherin in the TFV + anti-miR-4429 group were decreased (P< 0. 05), while the number of colonies formed and the number of invasive cells were increased (P< 0. 05), the wound healing rate and the expression level of N-cadherin were increased (P< 0. 05). Conclusion Total flavonoids from Valeriana jatamansi Jones can inhibit gastric cancer AGS cells proliferation, migration and invasion, and its anti-tumor mechanism may be related to the up-regulation of miR-4429 expression.

Piezo2, as a member of Piezo channel family, is a fast adaptive and mechanically activated cation channel expressed in the sensory neuron subsets of dorsal root ganglion. It can convert mechanical stimulation into mechanical sensitive current to control the generation of body surface proprioception, touch and pain. Nowadays, more and more studies show that Piezo2 plays an important role in the generation and transmission of a variety of pains. Based on the analysis of previous research results, this paper summarizes the structure and function of mechanically sensitive ion channel Piezo2 and its participation in different pains and the associated diseases, so as to understand the application prospects of using it as a potential intervention target for the treatment of pain.

Multiple myeloma is the second most common hematological malignancy, there is still no cure for it despite the great progress on its treatment in decades. This is found to be closely related to the cellular and non-cellular components in the immune microenvironment. For instance, the mesenchymal stromal cells in the immune microenvironment could mediate the immune escape, increase the osteoclast activity, and impaire the osteoblast differentiation. In addition, some immune cells and cytokines with abnormal immune function could promote the proliferation, survival and drug resistance of myeloma cells. In recent years, many new therapeutic tools has emerged. Therefore, an in-depth study of the interaction between the above components and myeloma cells is expected to provide some new ideas and strategies for the clinical treatment of multiple myeloma.