Objective To explore the effect of miR-133b on the cell proliferation and invasion of renal cell carcinoma ( RCC) by targeting polypyrimidine tract binding protein 1 ( PTBP1 ). Methods The expression level of miR-133b in RCC tissues and cells were detected. The targeting relationship between miR-133b and PTBP1 was verified. The effect of miR-133b on proliferation and migration of RCC cells were detected. Results The expression level of miR-133b in RCC tissues was decreased, while PTBP1 was up-regulated when compared with para-carcinoma tissues (P < 0. 05). PTBP1was the target gene of miR-133b. The colony formation rate, the number of invasive cells and the expression level of PTBP1 were significantly decreased in miR-133b mimic group, but were significantly increased in miR-133b inhibitor group, when compared with miR-NC group. The colony formation rate, the number of invasive cells and the expression level of PTBP1 were increased in miR-133b mimic + pc-PTBP1 group when compared with miR-133b mimic + pc-NC group (P< 0. 05). Conclusion miR-133b expression is down-regulated in RCC tissues and cells, which may affect cell proliferation and invasion by the regulation of PTBP1.

Objective To explore the effect of miR-766-5p on the proliferation, invasion and apoptosis of glioma cells. Methods The targeting relationship between miR-766-5p and matrix metalloproteinase-7 (MMP-7) was predicted by Targetscan, and then was verified. U87 cell lines with miR-766-5p overexpression, MMP-7 overexpression or miR-766-5p + MMP-7 overexpression were constructed. BrdU staining, flow cytometry, and transwell assay were used to assay the cell proliferation, apoptosis, and invasion. The expression levels of related proteins were detected by Western blotting. Results MiR-766-5p could inhibit the expression of MMP-7. Overexpression of miR-766- 5p could inhibit the expression of Ki67, proliferating cell nuclear antigen (PCNA), Wnt1 and β-catenin, and inhibit the proliferation and invasion of U87 cells. It could also promote the expression of Bcl-associated X protein (Bax) / B-cell lymphoma / leukemia-2 (Bcl-2), and promote cell apoptosis in U87 cells. Overexpression of MMP-7 could partially reverse the effect of miR-766-5p overexpression on U87 cells proliferation, invasion, and apoptosis. Conclusion MiR-766-5p can inhibit the proliferation and invasion of U87 cells, and promote the apoptosis of U87 cells by inhibiting the expression of MMP-7. 

Objective To investigate the effect of lncRNA HOXB-AS3 on cell proliferation, apoptosis and invasion in acute myeloid leukemia (AML) and the underlying mechanism. Methods The expression level of lncRNA HOXB-AS3 in bone marrow mononuclear cells ( BMMCs) of AML patients and the AML cell lines were detected by RT-qPCR. THP1 and HL60 cell lines stably expressed shRNA-HOXB-AS3-01, shRNA-HOXB-AS3-02 or shRNA-HOXB-AS3-03 were established by lentivirus transfection. CCK-8 assay, EdU assay, flow cytometry, and transwell assay were used to measure the cell viability, proliferation, apoptosis and invasion, Western blotting was employed to detect the protein levels. The AML nude mouse xenograft model was established to verify the effect of low expression level of HOXB-AS3 on the tumor growth in vivo. Results The expression level of lncRNA HOXB-AS3 in AML BMMCs and cell lines was increased significantly. The low expression of HOXB-AS3 significantly inhibited the cell viability, proliferation, and invasion of THP1 and HL60 cells, enhanced the apoptosis, up-regulated the expression levels of cleaved caspase-3, cleaved caspase-9 and E-cadherin, and down-regulated the expression levels of N-cadherin, VEGF, PI3Kp85α and p-AKT. The low expression of HOXB-AS3 significantly inhibited the tumor growth, and down-regulated the expression level of PI3Kp85α and p-AKT In vivo. Conclusion lncRNA HOXB-AS3 aggravates the proliferation, apoptosis and invasion of AML cells by activating the PI3K-AKT pathway.

Objective To explore the effect of gnaphalium affine extract (GAE) on the anterior cruciate ligament transverse (ACLT) -induced osteoarthritis (OA) in rats. Methods A total of 90 SPF-level healthy male SD rats were randomly divided into 6 groups (15 rats in each group): sham group, model group, GAE low-dose group (400 mg / kg), GAE medium-dose group (800 mg / kg), GAE high-dose group (1 600 mg / kg), and diacerein group (54 mg / kg). OA Rat model was established in above groups by performing the ACLT surgery except in the sham group, GAE or diacerein treatment were given to the rats in the corresponding groups for 30 d. The pathological damage of knee cartilage tissues was observed by HE staining. Enzyme-linked immunosorbent assay was used to measure the levels of inflammatory factors in serum and synovial fluid. The levels of oxidative stress factors in synovial fluid were measured by colorimetry. The expression levels of protein kinase R-like endoplasmic reticulum kinase / eukaryotic translation initator factor-2α / C / EBP homologous protein (PERK/ eIF2a / CHOP) signaling pathway related proteins in knee tissues were detected by Western blotting. Results The rat cartilage surfaces in the sham group were smooth and intact. The rat cartilage tissues in the model group and GAE low-dose group were severely damaged, the stratified structures of the articular cartilages were blurred and cell arrangements were disordered. The pathological damages in rats from the GAE medium-and high-dose groups and the diacerein group were significantly relieved compared with those from the model group. The Mankin’s score of rats was higher in the model group. The levels of serum and synovial fluid tumor necrosis factor-α (TNF-α), interleukin-6 ( IL-6 ) and interleukin-1β ( IL-1β) in the model group were higher than those in the sham group (P < 0. 05). The level of synovial fluid malondialdehyde (MDA) in the model group was higher than that in the sham group (P< 0. 05), while the levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were lower (P < 0. 05). The expression levels of PERK, eIF2a and CHOP were higher in the model group than in the sham group (P < 0. 05). The Mankin’s score of the rats in the GAE medium-and high-dose groups and the diacerein group was lower. The levels of serum and synovial fluid TNF-α, IL-6 and IL-1β in the GAE medium-and high-dose groups and the diacerein group were lower than those in the model group (P < 0. 05). The synovial fluid MDA level in the GAE medium-and high-dose groups and the diacerein group was lower (P< 0. 05), while the levels of SOD and GSH-Px were higher than those in the model group (P< 0. 05). The expression levels of PERK, eIF2a and CHOP were lower in the GAE medium-and high-dose groups and the diacerein group than in model group (P< 0. 05). Conclusion An appropriate dosage of GAE can significantly improve the pathological damage of knee joints, reduce the inflammatory response and oxidative stress, and inhibit the activity of PERK/ eIF2a / CHOP signaling in the OA rats. 

Objective To explore the effect of gypenosides ( GP ) on the osteoarthritis rats. Methods The experimental rats were randomly divided into sham group, model group, low-dose, medium-dose and high-dose GP groups, and diclofenac group. The pathological damage of the cartilages was detected by HE staining and alician blue staining. The BV/ TV, Tb. N, Tb. Sp and Tb. Th values were measured by CT. The expression levels of cartilage matrix related proteins and SIRT1 / AMPK were detected by Western blotting. The inflammatory factors and oxidative stress indexes levels in cartilage tissues and serum were determined by ELISA. Results The inflammatory infiltration and fibrosis of cartilage tissues were improved significantly with an increased treatment of GP. The BV/ TV, Tb. Th and Tb. N values were significantly increased, while the Tb. Sp value was significantly decreased (P < 0. 05). The expression levels of SOX-9, Aggreca and Collagen Ⅱ in the cartilage matrix were significantly increased (P< 0. 05). The levels of TNF-α, IL-6 and IL-1β in serum and cartilage tissues were significantly decreased (P< 0. 05). The levels of SOD and GSH-Px in cartilage tissues were significantly increased, while the MDA level was significantly decreased (P< 0. 05). The expression levels of p-AMPK, SITR1 and PGC1α in cartilage cells were significantly increased (P< 0. 05). Conclusion GP can reduce the oxidative stress and repair the cartilage damage by activating AMPK/ SITR1. 

Objective This study was designed to evaluate the anti-inflammatory effect and mechanisms of SAI and SAK (SA-A and SA-C) in vitro. Methods RAW264. 7 cells were stimulated by lipopolysaccharide (LPS) to establish an in vitro inflammatory model. Cells were divided into groups as follows: normal control group, model control group, SAI and SAK groups (0. 4-40 μg· mL ^{- 1} ). Cell viability was measured. The nitric oxide (NO) release, intracellular calcium concentration and ROS level were measured. The expression levels of iNOS and COX-2 mRNA were detected. The effect of SAI and SAK on the above indicators were evaluated. Results LPS can induce NO release in the supernatant of RAW264. 7 cells, increasing the Ca ^{2 +} concentration and the ROS level in cells. A concentration of 10 μg·mL ^{- 1} of SAI or SAK significantly inhibited the above indicators, and 10 μg·mL ^{- 1} of SAK was more effective than that of SAI. Also, SAI and SAK inhibited LPS-induced up-regulation of iNOS and COX-2 mRNA expressions in RAW264. 7 cells. Conclusion Both of SAI and SAK can inhibit the LPS-induced inflammatory response in RAW264. 7 cells. The anti-inflammatory mechanism is related to the inhibition of iNOS and COX-2 mRNA expressions. Moreover, SAK has more anti-inflammatory activity after removing the toxic SA-B and SA-D contents when compared with SAI. This study has certain significance for the drug production.

Objective To study the effect of transcription factor 7-like 2 (Tcf7l2) on the adipogenic differentiation of bone marrow mesenchymal stem cells ( BMSCs) and the regulation of Wnt / β-catenin signaling pathway in BMSCs. Methods Tcf7l2 shRNA and overexpression lentivirus were made and transfected into the BMSCs with or without adipogenesis to construct the sh-Tcf7l2 and Tcf7l2-overexpression BMSCs. QRT-PCR was used to detect the expression level of Tcf7l2 mRNA in BMSCs. Oil red O staining was used to assess the adipogenic differenciation of BMSCs. Western blotting was used to detect the expression levels of Wnt / β-catenin signaling pathway downstream proteins β-catenin, c-myc and cyclin D1 and some other associate proteins. Results The expression levels of Tcf7l2 mRNA, β-catenin, c-myc and cyclin D1 proteins were reduced, and the number of fat particles, the expression levels of PPAR-γ and C / EBP-α were increased in the sh-Tcf7l2 group when compared with the control groups (P < 0. 05) . The expression levels of Tcf7l2 mRNA, β-catenin, c-myc and cyclin D1 proteins were increased, and the number of fat particles, the expression levels of fat-related proteins PPAR-γ and C / EBP-α were reduced in the Tcf7l2-overexpression group, when compared with the control groups (P < 0. 05) . Conclusion Tcf7l2 overexpression can inhibit the adipogenic differentiation of BMSCs, the mechanism of which may be related to the activation of Wnt / β-catenin signaling pathway. 

Objective To explore the effect of cytokeratin 19 on the proliferation, invasion and metastasis of thyroid cancer cells and the related mechanisms. Methods In this study, human thyroid cancer cells (B-CPAP) were used as the research object. MTT assay, wound healing assay and cell invasion assay were applied to investigate the effect of cytokeratin 19 on the cell proliferation, invasion and metastasis after the treatment of keratin 19 on B-CPAP. Western blotting was used to detect the related protein expressions to explore the underlying mechanisms. Results MTT assay showed that the proliferation rate of B-CPAP increased significantly with the increasing concentration of keratin 19 treatment. Transwell assay and wound healing assay results showed that keratin 19 can promote the migration ability of B-CPAP in a dose-dependent manner significantly. In addition, PTEN and MAPK pathways may play roles in that process. Conclusion Cytokeratin 19 can significantly increase the proliferation, invasion and metastasis of B-CPAP, and this effect of keratin 19 is related to PTEN and MAPK pathways.

Objective To investigate the effect of diosgenin on the oral squamous cell carcinoma HSC4 cells. Methods HSC4 cells were divided into four groups: diosgenin 0, 0. 5, 1 and 2 μmol / L. Cell proliferation, apoptosis, migration and invasion were assayed by CCK-8, 5-bromo2′-deoxyuridine (BrdU) staining, colony formation experiment, flow cytometry, wound healing assay and transwell assay. The protein expression levels of Bcl-2, Bax and cleaced caspase-3 were detected by Western blotting. JC-1 was employed to measure the mitochondrial membrane potential. H2DCFDA was used to detect the production of reactive oxygen species (ROS). GSH and MDA levels were detected by the kits. Results The cell viability, colony formation ability, BrdU positive cell rate, cell migration and invasion ability, mitochondrial membrane potential, GSH level, and the expression level of Bcl-2 were decreased in the 1 μmol / L and 2 μmol / L diosgenin groups when compared with the 0 μmol / L diosgenin group. The apoptosis rate, the expression levels of Bax and cleaced caspase-3, and the ROS and MDA levels were increased in the 1 μmol / L and 2 μmol / L diosgenin groups when compared with the μmol / L diosgenin group. Conclusion Diosgenin can inhibit the proliferation and motility of HSC4 cells, and induce apoptosis through mitochondrial pathway and ROS production.

Objective To investigate the value of rifampicin resistant real-time fluorescent quantitative nucleic acid amplification technology (GeneXpert MTB / RIF) combined with the gene chip technology in the diagnosis of smear-negative Mycobacterium tuberculosis (MTB) and the clinical significance of serum soluble triggering receptor-1 (sTREM-1) and procalcitonin (PCT) levels in pulmonary tuberculosis. Methods A total of 130 cases of suspected smear-negative pulmonary tuberculosis patients in Liaocheng People’s Hospital from January 2019 to January 2021 were selected. Alveolar lavage fluid were collected, GeneXpert MTB / RIF were performed, gene chip method were applied, and the serum levels of sTREM-1 and PCT were detected. The alveolar lavage fluid tuberculosis culture method and the drug sensitivity test were used as the gold standard to evaluate the diagnostic value of GeneXpert MTB / RIF combined with gene chip technology for the smear-negative pulmonary tuberculosis. ROC analysis was used to evaluate the diagnostic value of serum sTREM-1 and PCT in smear-negative pulmonary tuberculosis. Results A total of 58 patients with non-smear-negative pulmonary tuberculosis and 72 patients with smear-negative pulmonary tuberculosis were diagnosed by using the alveolar lavage fluid tuberculosis culture method. The sensitivity of GeneXpert MTB / RIF combined with gene chip technology for the diagnosis of pulmonary tuberculosis was 68. 06 % , which was higher than that of gene chip method alone (P< 0. 05). The specificity, accuracy, positive predictive value and negative predictive value of GeneXpert MTB / RIF combined with gene chip method for the diagnosis of pulmonary tuberculosis were 91. 38 % , 78. 46 % , 90. 74 % and 69. 74 % , respectively, despite that the differences were not statistically significant when compared with the GeneXpert MTB / RIF or gene chip method alone (P> 0. 05). The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of GeneXpert MTB / RIF combined with gene chip method for the diagnosis of MDR-TB were 94. 12 % , 85. 45 % , 87. 50 % , 66. 67 % and 97. 92 % , respectively, compared with GeneXpert MTB / RIF and gene chip method alone, the differences were not statistically significant (P> 0. 05). The levels of serum sTREM-1 and PCT in patients with severe pulmonary tuberculosis were (18. 04 ± 2. 07) ng / mL and (2. 09 ± 0. 19) ng / mL respectively, which were significantly higher than those in patients with mild pulmonary tuberculosis ( P < 0. 05 ). The areas under the ROC curve of serum sTREM-1 and PCT for the prediction of severe pulmonary tuberculosis were 0. 811 and 0. 844. The cut-off values were 16. 32 ng / mL and 2. 00 ng / mL, the sensitivities were 89. 30 % and 82. 00 % , the specificities were 61. 40 % and 79. 60 % , respectively. Conclusion The GeneXpert combined with gene chip technology can improve the diagnostic sensitivity of smear-negative MTB, and both methods have good diagnostic values for MDR-TB. The levels of sTREM-1 and PCT in patients with severe pulmonary tuberculosis were significantly higher than those in patients with mild pulmonary tuberculosis. Serum sTREM-1 and PCT have high sensitivities and specificitise in the diagnosis of severe pulmonary tuberculosis, which can be applied for the early diagnosis of pulmonary tuberculosis.

Objective To study the regulatory effect of arginine regulatory protein ArgR on the gene expression of type Ⅵ secretion system (T6SS) in P. aeruginosa (Pseudomonas aeruginosa). Methods The ArgR protein expression vector with SUMO tag was constructed, and the ArgR protein of P. aeruginosa was expressed heterologously by using E. coli DH5α. The gel electrophoretic mobility shift assay (EMSA) was used to explore the interaction between ArgR and the promoter of T6SS-related gene hsiA2. Hcp2-Flag expression vector with FLAG tag was constructed. Western blotting and qRT-PCR assays were used to detect the regulatory effect of argR gene on the T6SS structural gene hcp2. Results The soluble ArgR protein was obtained by cutting off the label by restriction enzymes. EMSA analysis showed that there was an interaction between ArgR protein and the hsiA2 promoter. The results of Western blotting and qRT-PCR showed that the expression level of hcp2 was significantly increased in the mutants lacking argR. Moreover, the expression level of hcp2 in the mutants could be returned to an equivalent level as in the wild-type bacteria after argR expression level was supplemented. Conclusion This study shows that ArgR can bind to the promoter of hsiA2 and negatively regulate the expression of T6SS-related genes. 

Diabetic angiopathy is the main cause of disability and death in diabetes. However, strategies for the prevention and treatment of diabetic angiopathy have not achieved ideal results. Mechanistic target of rapamycin kinase ( mTOR) is a serine / threonine kinase presented in eukaryotes and plays an important role in various biological processes. A large number of studies have found that mTOR could participate in the occurrence and progress of diabetic angiopathy by regulating cell proliferation, apoptosis, autophagy, oxidative stress and inflammatory responses. This article reviews the effect and related mechanisms of mTOR on diabetic atherosclerosis, diabetic nephropathy and diabetic retinopathy, and provides a new insight for the prevention and treatment of diabetic angiopathy. 

Acyl-CoA synthetase long chain family (ACSLs) catalyzes the connection of CoA with 12-20 long chain fatty acids and participates in lipid synthesis and metabolism. ACSL5 is a subtype of ACSLs that is both located on mitochondria and involved in intestinal epithelial cell apoptosis, and is expressed in a variety of tissues. Studies on different cancer types have reported that ACSL5 expression in cancer patients is significantly different from that in healthy individuals, and it has been proved that high expression level of ACSL5 has a positive effect on the prognosis of patients with lung cancer, pancreatic cancer, breast cancer and ovarian cancer. This paper aims to review on the advances in the effect of ACSL5-meidated lipid metabolism on cancer, which may provide a new strategy for cancer therapy via targeting the lipid metabolism enzymes.

Renal cell carcinoma (RCC) affects all human beings profoundly and extensively. Clear cell renal cell carcinoma ( ccRCC) is the most common pathological subtype of RCC. In the past 30 years, the biomolecular mechanisms of RCC have been continuously explored. With the in-depth research on DNA repair pathways, the correlates between RCC and the DNA repair genes were gradually discovered. Cells were always exposed to various factor which could lead to the genome damages, and the DNA repair system helps to maintain the genome stability in that process. If the DNA repair system is defective, the genome will become unstable, cells will undergo apoptosis or transform into cancer cells. This review focuses on the important biological correlates between DNA repair genes (DRG) and ccRCC in the susceptibility, prognosis, and treatment of ccRCC. It could help lay a certain foundation for the further exploration of molecular mechanisms between DNA damage repair pathways and ccRCC, the determination of new ccRCC molecular markers, and the finding of suitable immunosuppressive therapeutic targets.