OBJECTIVE: To investigate the effects of 4-methylcatechol (4MC) on the migration and inflammatory response in rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS), as well as its underlying mechanisms of action. METHODS: RA-FLS was isolated from synovial tissue donated by RA patients, and the optimal concentration of 4MC was determined by cell counting kit 8 method for subsequent experiments, and the effect of 4MC on the migratory ability of RA-FLS was evaluated via a cell scratch assay. An inflammation model of RA-FLS was induced by tumor necrosis factor α (TNF-α). Real-time fluorescence quantitative PCR and ELISA were employed to detect the gene and protein expression levels of interleukin-1β (IL-1β) and IL-6 in RA-FLS and their culture supernatants, respectively, thereby investigating the anti-inflammatory effects of 4MC. Western blot was used to examine the expressions of nuclear factor κB (NF-κB) signaling pathway-related proteins, including inhibitor of NF-κB-α (IKBα), phosphorylated (P)-IκBα, NF-κB-inducing kinase α (IKKα), P-IKKαβ, P-p65, and p65. Cellular immunofluorescence was utilized to detect the expression and localization of p65 in RA-FLS, exploring whether 4MC exerts its anti-inflammatory effects by regulating the NF-κB signaling pathway. Finally, a collagen-induced arthritis (CIA) mouse model was established. The anti-RA effect of 4MC in vivo was evaluated by gross observation and histological examination. RESULTS: 4MC inhibited RA-FLS migration in a concentration-dependent manner. In the TNF-α-induced RA-FLS inflammation model, 4MC significantly decreased the gene and protein expression levels of IL-1β and IL-6. Furthermore, 4MC markedly reduced the ratios of P-IΚBα/IΚBα, P-IKKαβ/IKKα, and P-p65/p65, thereby blocking the transcriptional activity of p65 by inhibiting its nuclear translocation. This mechanism effectively suppressed the activation of the TNF-α-mediated NF-κB signaling pathway. Animal studies demonstrated that 4MC [10 mg/(kg·day)] significantly lowered serum levels of IL-1β, IL-6, and TNF-α, and alleviated arthritis severity and bone destruction in CIA mice. CONCLUSION 4MC not only inhibits the migration of RA-FLS but also mitigates their inflammatory response by suppressing the NF-κB signaling pathway, thereby effectively exerting its anti-RA effects.
Synoviocytes/metabolism* ; Arthritis, Rheumatoid/metabolism* ; Animals ; Cell Movement/drug effects* ; Humans ; Catechols/therapeutic use* ; Fibroblasts/drug effects* ; Mice ; Tumor Necrosis Factor-alpha/pharmacology* ; Interleukin-1beta/metabolism* ; Interleukin-6/metabolism* ; Signal Transduction/drug effects* ; NF-kappa B/metabolism* ; Transcription Factor RelA/metabolism* ; Synovial Membrane/cytology* ; Cells, Cultured ; Male ; Arthritis, Experimental ; Anti-Inflammatory Agents/pharmacology* ; NF-KappaB Inhibitor alpha ; Inflammation

With the development of high-throughput sequencing technology, circular RNAs (circRNAs) have gradually become a hotspot in the research on non-coding RNA. CircRNAs are produced by the covalent circularization of a downstream 3' splice donor and an upstream 5' splice acceptor through backsplicing, and they are pervasive in eukaryotic cells. CircRNAs used to be considered byproducts of false splicing, whereas an explosion of related studies in recent years has disproved this misconception. Compared with the rich studies of circRNAs in animals, the study of circRNAs in plants is still in its infancy. In this review, we introduced the discovery of plant circRNAs, the discovery of plant circRNAs, the circularization feature, expression specificity, conservation, and stability of plant circRNAs and expounded the identification tools, main types, and biogenesis mechanisms of circRNAs. Furthermore, we summarized the potential roles of plant circRNAs as microRNA (miRNA) sponges and translation templates and in response to biotic/abiotic stress, and briefed the degradation and localization of plant circRNAs. Finally, we discussed the challenges and proposed the future directions in the research on plant circRNAs.
Animals ; MicroRNAs/metabolism* ; Organelle Biogenesis ; Plants/metabolism* ; Protein Biosynthesis/physiology* ; RNA, Circular/metabolism* ; RNA, Plant/metabolism* ; Research/trends* ; Stress, Physiological/genetics*

Apomixis has been widely concerned because of its great potential in heterosis fixation. Artificial apomixis is an important direction of current apomixis research. Mitosis instead of Meiosis (MIME) produces diploid gametes that is identical with the maternal genetic composition and is a key step in the artificial creation of apomixes. This paper reviews the occurrence of MIME and its application in crop apomixis and the problems encountered, in an aim to provide reference for expanding the application of MIME in crop apomixis.
Apomixis ; Crops, Agricultural ; genetics ; Diploidy ; Germ Cells ; Meiosis ; Mitosis

Objective To investigate the molecule mechanism of microRNA (miR)-138 in inhibiting invasion and migration of breast cancer by regulating epithelial mesenchymal transformation (EMT). Methods Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect expression of miR-138 after transfecting miR negative control simulacrum (miR-NC) and miR-138 simulacrum in human normal mammary epithelial cell (MCF-10A) and breast cancer cells (MCF-7 and MDA-MB-231) from July 2017 to June 2018. MTT method was used to detect the breast cancer cell activity. Cell scratch test and Transwell test were used to detect the breast cancer cell migration distance and invasion rate. RT-PCR was used to detect the expression of the EMT key molecules Vimentin, N-cadherin and E-cadherin after transfecting miR-138 simulacrum. Results The expression level of miR-138 in MCF-10A was significantly higher than that in MCF-7 and MDA-MB-231 (1.006 ± 0.009 vs. 0.324 ± 0.027 and 0.512 ± 0.068), and there was statistical difference (P<0.05);there was no statistical difference in the expression level of miR-138 between MCF-7 and MDA-MB-231 (P>0.05). The breast cancer cell viabilities of MCF-7 and MDA-MB-231 at third and fourth day after transfecting miR-138 simulacrum were significantly lower than those of transfecting miR-NC (MCF-7: 0.514 ± 0.052 vs. 0.593 ± 0.061 and 0.643 ± 0.074 vs. 0.784 ± 0.081;MDA-MB-231:0.552 ± 0.043 vs. 0.614 ± 0.063 and 0.673 ± 0.074 vs. 0.792 ± 0.077), and there were statistical differences (P<0.05). The breast cancer cell migration distances and invasion rates of MCF-7 and MDA-MB-231 after transfecting miR-138 simulacrum were significantly lower than those of transfecting miR-NC (MCF-7: 0.572 ± 0.051 vs. 1.003 ± 0.012 and 0.624 ± 0.043 vs. 1.002 ± 0.007, MDA-MB-231:0.472 ± 0.051 vs. 1.003 ± 0.095 and 0.573 ± 0.044 vs. 1.004 ± 0.091), and there were statistical differences (P<0.05). The expressions of Vimentin and N-cadherin mRNA in MCF-7 and MDA-MB-231 after transfecting miR-138 simulacrum were significantly lower than those of transfecting miR-NC, but the expression of E-cadherin mRNA was significantly increased, and there were statistical differences (P<0.05). Conclusions The expressions of miR-138 in both breast cancer cells decreased. Overexpression of miR-138 in breast cancer cell can inhibit proliferation, migration and invasion via regulating EMT.

For the last decade,the incidence of kidney neoplasms has shown an obvious rising trend in the world. The most common histopathological type of kidney neoplasms is clear cell renal cell carcinoma (ccRCC),which has a poor prognosis. ccRCC is generally accompanied by reprogramming of glucose,fatty acid,glutamine,tryptophan and arginine metabolic networks and pathways. Reprogramming of metabolic net-works and pathways enables tumor cells to proliferate rapidly,survive in conditions of nutrient depletion and hy-poxia,and escape surveillance by epidemic systems. New strategies have been developed to the treatment of ccRCC by targeting key proteins or enzymes involved in metabolic reprogramming pathways.

Objective To investigate the effects of microRNA-134 (miR-134) on the proliferation and apoptosis of non-small cell lung cancer (NSCLC) and its potential molecular mechanism.Methods Quantitative real-time fluorescent polymerase chain reaction (qRT-PCR) was used to detect the differences of miR-134 expression between 10 cases of lung cancer tissues and normal lung tissues,and between normal human lung epithelial cell line BEAS-2B and lung adenocarcinoma cell line A549.miR-NC and miR-134 mimic were transfected into A549 cells.The effect of miR-134 on proliferation of A549 cells was detected by methyl thiazolyl tetrazolium (MTT) and colony form experiment.Flow cytometry was used to determine the effect of miR-134 on A549 cells apoptosis.The effect of miR-134 on the expression of P53 protein was detected by Western blotting.Results The relative expressions of miR-134 in NSCLC tumor tissues and adjacent tissues were 0.429 ± 0.126 and 0.971 ±0.183 respectively,and the difference was statistically significant (t =7.742,P ＜0.001).The relative expressions of miR-134 in BEAS-2B cells and A549 cells were 1.013 ± 0.095 and 0.371 ± 0.068 respectively,and the difference was statistically significant (t =17.377,P ＜ 0.001).The absorbance (A) values of A549 cells transfected with miR-mimic were 0.451 ±0.051 and 0.518 ±0.074 on the third and forth day respectively,and those of A549 cells transfected with miR-NC were 0.683 ± 0.041 and 0.815 ± 0.065 respectively.The proliferation ability of miR-mimic group was significantly lower than that of miR-NC group (t =12.965,P ＜ 0.001;t =9.535,P ＜ 0.001).The colony forming rates of A549 cells transfected with miR-NC and miR-134 mimic were 91.2％ ± 8.3％ and 38.6％ ±4.5％ respectively,and the colony forming rate of A549 cells in miR-134 mimic group was significantly decreased (t =17.617,P ＜0.001).The apoptosis rates of miR-134 mimic group and miR-NC group were 93.5％ ± 3.7％ and 85.4％ ± 2.0％ respectively,and the difference was significant difference (t =6.119,P ＜ 0.001).The relative expressions of P53 protein in miR-134 mimic group and miR-NC group were 1.816 ±0.173 and 0.992 ± 0.096 respectively,and the difference was statistically significant (t =19.308,P ＜ 0.001).Conclusion miR-134 can be an effective target for the treatment of NSCLC by increasing the protein expression of P53,inhibiting the viability and proliferation of tumor cells,and promoting the apoptosis of tumor cells.

BACKGROUND: The epidermal growth factor receptor (EFGR) mutation was closely related to the invasion and metastasis of lung adenocarcinoma and the biological axis of CXCR4/CXCL12 (chemokine receptor 4/chemokine ligand 12) played an important role in the organ-specific metastasis of the tumor. It was a question surrounding whether there is interaction between them in the process of lung adenocarcinoma metastasis. To investigate the potential molecular mechanisms of EGFR over-expression and EFGR-mutations effects on cell proliferation, migration and invasion, we constructed EGFR over-expression and three EFGR-mutant human lung adenocarcinoma H1299 cell sublines. METHODS: EGFR over-expression and three EFGR-mutant (EGFR-E746-A750del, EGFR-T790M and EGFR-L858R) plasmid were designed and transfected H1299 cells with Lipofectamine 2000. H1299 cells transfected with empty vector were negative control (NC), and H1299 cells without transfection were set as blank control (BC). The effects of EGFR over-expression and mutations on the proliferation, migration and invasion of H1299 cells were detected by cell cloning assay, wound healing assay and Transwell assay. The mRNA and protein expression levels of MMP-2, MMP-9, CXCR4 and CXCL12 were detected by RT-PCR and Western blot. RESULTS: Compared with negative control group and blank control group, EGFR over-expression and EGFR-E746-A750 deletion have significantly higher colony formation (28±2, 28.33±4.16; respectively) (P<0.05) and the cell migration and invasion ability were significantly increased (P<0.05). RT-PCR and Western blot assay showed that the mRNA and protein expression of MMP-2, MMP-9, CXCR4 and CXCL12 in EGFR over-expression and EGFR-E746-A750 deletion group were remarkably higher than that in negative control and blank control group (P<0.05). CONCLUSIONS EGFR over-expression and 19 exon deletion can promote the expression of MMP-2 and MMP-9 by up-regulating CXCR4/CXCL12 signaling pathway, leading to the change of tumor biological characteristics such as higher proliferation, migration and invasion ability.
Adenocarcinoma ; pathology ; Adenocarcinoma of Lung ; Cell Line, Tumor ; Cell Movement ; genetics ; Chemokine CXCL12 ; metabolism ; ErbB Receptors ; genetics ; Gene Expression Regulation, Neoplastic ; Humans ; Lung Neoplasms ; pathology ; Mutation ; Neoplasm Invasiveness ; Receptors, CXCR4 ; metabolism ; Signal Transduction ; genetics

Objective:To explore the influence of ulinastatin combined with thymosinα1 on the immune function of patients with a-cute brain injury. Methods:Sixty-eight cases of patients with acute brain injury were divided into the control group and the observation group randomly with thirty-four ones in each. The control group was given the routine treatment, and the observation group was given ulinastatin combined with thymosinα1 additionally. After the 1-day, 3-day, 7-day and 14-day treatment, transforming growth factor-β1 (TGF-β1), interleukin-6(IL-6), interleukin-10 (IL-10), tumor necrosis factor-α (TNF-α) and the other serum inflammatory cyto-kine levels, and CD4 +, CD8 +, CD4 + /CD8 +, HLA-DR and cellular immune index levels were detected in the two groups. The prog-nosis effects were evaluated by the prognostic classification of brain injury, and the adverse reactions were analyzed in the two groups as well. Results:After the 1-day treatment, there were no significant differences in the serum inflammatory cytokines and immune param-eters between the groups (P>0. 05). After the 3-day, 7-day and 14-day treatment, serum TGF-β1, IL-6, IL-10 and TNF- α levels were higher than those on the first day after the treatment, and TGF-β1 showed an increasing trend with time extension, while IL-6, IL-10, TNF-α and CD4 + and CD4 + /CD8 +rose first and then decreased. After the 3-day, 7-day and 14-day treatment, serum IL-10 and CD4 +levels in the observation group were significantly higher than those in the control group, and IL-6 and TNF-αlevels were signifi-cantly lower than those in the control group (P<0. 05). After the 3-day and 14-day treatment, CD4 + and CD8 + levels in the observa-tion were significantly higher than those in the control group, and after the 7-day and 14-day treatment, HLA-DR levels were signifi-cantly higher than those in the control group (P<0. 05). The prognosis effect of the observation group was better than that of the con-trol group with statistically significant difference (P<0. 05). Conclusion:Ulinastatin combined with thymosin α1 is used to treat the patients with acute brain injury with better cellular immune function improvement and prognosis effect, which is worthy of clinical popu-larization and application.

Objective To explore the effectiveness of epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC) patients with EGFR double mutations containing rare mutations.Methods From March 2007 to January 2017,NSCLC patients with EGFR double mutations containing rare mutations confirmed by histopathology and EGFR mutation detections at Renmin Hospital of Wuhan University,Zhongnan Hospital of Wuhan University and Hubei Cancer Hospital were enrolled.According to the mutation types,patients were divided into double activating mutations group and activating mutations with insensitive mutations group.The effectiveness of TKIs in NSCLC patients with EGFR double mutations containing rare mutations and different mutation types was analysis.Results Among the 2 637 NSCLC patients underwent EGFR mutation detections,19 patients were confirmed as EGFR double mutations containing rare mutations.Fifteen patients received EGFR-TKIs therapy,the objective response rate (ORR),disease control rate (DCR) and median progression free survival (PFS) were 46.7％ (7/15),73.3％ (11/15) and 8.1 months,respectively.In patients with double activating mutations,two patients had partial response (PR),one patient had a stable disease (SD).In the activating mutations with insensitive mutations group,five patients had PR,three patients had SD,four patients had no effect.Conclusion Patients with EGFR double mutations containing rare mutations have a general response to EGFR-TKIs,and this result can provide a reference for the treatment of those patients.

Objective To study the risk factors for pathological upgrading after diagnosis of esophageal low?grade intra?epithelial neoplasia with ESD preoperative biopsy. Methods The endoscopic and pathological data of 85 lesions with ESD preoperative biopsy were analyzed, and grouped based on pathological upgrading after ESD. The risk factors for pathological upgrading after ESD was studied through single and multiple factor analysis. Results Pathological upgrading occurred in 45(52?94%) lesions after ESD, among whom 38 lesions developed up to high?grade intra?epithelial neoplasia and 7 lesions developed to esophageal early cancer. NBI?ME was performed on 37 patients and the accuracy of detecting the pathological invasion was 83?8%(31/37).Multi?factor analysis showed that reddish surface(OR=9?478, 95%CI:2?775?32?368, P = 0?000 3 ) and nodular lesion ( OR = 15?628, 95%CI:1?475?165?617, P =0?022 5) were independent factors for pathological upgrading after ESD. Conclusion Pathological upgrading of low?grade intra?epithelial neoplasia was common, especially esophageal mucosa with red surface and nodular lesion.Biopsy combined with NBI?ME is of significant importance to improve diagnostic accuracy.