Information feedback of 18F-FDG PET/CT computer imaging combined with tumor markers on recurrence and metastasis of non-small cell lung cancer.

To investigate the diagnostic value or information feedback of tumor markers combined with 18F-FDG PET/CT computer imaging on recurrence and metastasis of non-small cell lung cancer (NSCLC). METHODS: From January 2013 to December 2017, 95 NSCLC patients undergoing systemic 18F-FDG PET/CT computer imaging examination at the PET/CT computer imaging center of Mudanjiang Medical University had been enrolled. Typically, the interval between the completion of treatment and PET/CT computer imaging examination should be at least three months. Besides, all patients had undergone serum CEA monitoring before and after 18F-FDG PET/CT computer imaging, and 70 of them had received CYFRA21-1 test at the same time. Tumor markers were examined with PET/CT at intervals of less than one week, and all the feedback results were compared with clinical follow-up results or final pathology. Additionally, all the enrolled patients were followed up for 6-12 months. RESULTS: The sensitivity, accuracy, specificity, positive predictive value and negative predictive value of 18F-FDGPET/CT information feedback in evaluating recurrence or metastasis after NSCLC treatment were superior to those of common tumor markers, and the differences were statistically significant (P<0.05). Those of 18F-FDG PET/CT computer imaging combined with tumor marker examination for the recurrence and/or metastasis after NSCLC treatment were remarkably higher than those of either individual examination, and the accuracy difference of information feedback had significant statistical significance (P<0.05). Clearly, the diagnosis using tumor markers was correlated with that by 18F-FDG PET/CT imaging, and the correlation coefficient was r=0.63. Moreover, serum CEA was grouped at different levels, and the positive rate and accuracy of 18F-FDG PET/CT computer imaging diagnosis were increased with the increase in CEA level. 8 patients had received 18F-FDG PET/CT dual-phase examination, among them, 4 were diagnosed with recurrence or metastasis after MSCLC treatment, and all of them had been detected.

Expression, purification, and evaluation of in vivo anti-fibrotic activity for soluble truncated TGF-ß receptor II as a cleavable His-SUMO fusion protein.

Excessive production of transforming growth factor-ß1 (TGF-ß1) and its binding to transforming growth factor-ß receptor type II (TGF-ßRII) promotes fibrosis by activation of the TGF-ß1-mediated signaling pathway. Thus, the truncated extracellular domain of TGF-ßRII (tTßRII) is a promising anti-fibrotic candidate, as it lacks the signal transduction domain. In this work, the native N-terminal tTßRII was prepared as a His-SUMO fusion protein (termed His-SUMO-tTßRII) in Escherichia coli strain BL21 (DE3). His-SUMO-tTßRII was expressed as a soluble protein under optimal conditions (6 h of induction with 0.5 mM IPTG at 37 °C). His-SUMO-tTßRII was purified by Ni-NTA resin chromatography, and then cleaved with SUMO protease to release native tTßRII, which was re-purified using a Ni-NTA column. Approximately 12 mg of native tTßRII was obtained from a one liter fermentation culture with no less than 95% purity. In vivo studies demonstrated that tTßRII prevented CCl4-induced liver fibrosis, as evidenced by the inhibition of fibrosis-related Col I and α-SMA protein expression in C57BL/6 mice. In addition, tTßRII downregulated phosphorylation of SMAD2/3, which partly repressed TGF-ß1-mediated signaling. These data indicate that the His-SUMO expression system is an efficient approach for preparing native tTßRII that possesses anti-liver fibrotic activity, allowing for the large-scale production of tTßRII, which potentially could serve as an anti-fibrotic candidate for treatment of TGF-ß1-related diseases.

MiR-1290 promotes proliferation, migration, and invasion of glioma cells by targeting LHX6.

To investigate the interaction of miR-1290 and LHX6 in gliomas, and their influence on the propagation and metastasis of glioma cells. The differential expression of miR-1290 in glioma cells was identified by chip screening. The expression level of miR-1290 and LHX6 were determined by qRT-PCR and Western blot. The influence of miR-1290 on propagation of glioma cells were analyzed by MTT assay, EdU incorporation, and colony formation, while the impact of miR-1290 on metastasis was assessed by transwell assay. The relationship between LHX6 and miR-1290 was testified by luciferase reporter assay. The gliomas orthotopic implantation model of nude mice was established to investigate the influence of miR-1290 and LHX6 on tumor growth. Tumor volumes were evaluated by photon density, and the expression of Ki67 protein was analyzed by immunohistochemistry. MiR-1290 presented a higher expression in glioma cells and tissues. MiR-1290 overexpression significantly promoted propagation and metastasis of glioma cells, while miR-1290 knockdown inhibited glioma development. MiR-1290 suppressed LHX6 expression, facilitating development of glioma cells. The orthotopic implantation model showed that miR-1290 overexpression promoted tumor growth while LHX6 overexpression inhibited it. MiR-1290 could promote glioma cell propagation and metastasis by inhibiting LHX6.

MiR-129-5p influences the progression of gastric cancer cells through interacting with SPOCK1.

The purpose of our study is to clarify the effect of microRNA-129-5p in the progression of human gastric cancer cells by regulating SPOCK1. The expression of microRNA-129-5p and SPOCK1 was tested by quantitative real-time polymerase chain reaction in tissues and cell lines. We validated the targeted relationship between microRNA-129-5p and SPOCK1 by dual luciferase reporter gene assay. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, colony formation, flow cytometry, transwell, and wound scratch assays were used to analyze the effects of microRNA-129-5p on SGC-7901 cell viability, proliferation, cell cycle and apoptosis, invasiveness, and migration. MicroRNA-129-5p was downregulated while SPOCK1 was upregulated in gastric cancer tissues and cell lines. The result of luciferase reporter gene assay demonstrated that microRNA-129-5p can target SPOCK1 by binding to the 3'untranslated region. The overexpression of microRNA-129-5p or the inhibition of SPOCK1 inhibited SGC-7901 viability, proliferation, migration, and invasion while promoted cell cycle arrest in G0/G1 stage and cell apoptosis. Our results suggested that microRNA-129-5p could directly specifically suppress SPOCK1, which might be one of the potential mechanisms in inhibiting cell processes including viability, proliferation, cell mitosis, migration, and invasiveness of gastric cancer cells.

Interaction between miR-572 and PPP2R2C, and their effects on the proliferation, migration, and invasion of nasopharyngeal carcinoma (NPC) cells.

We investigated the how miR-572 regulates PPP2R2C, and studied the effects of miR-572 and PPP2R2C on proliferation and migration as well as invasion of nasopharyngeal carcinoma (NPC) cells. NPC tissues and normal tissues were collected, and the expressions of miR-572 and PPP2R2C were detected by real-time PCR. Western blot was applied to detect the expression of PPP2R2C protein. The target relationship between miR-572 and PPP2R2C was confirmed by dual luciferase reporter gene assay. MTT assay and flow cytometry were applied to investigate the viability and apoptosis levels of NPC cells. Transwell as well as wound healing assays were used, respectively, to detect the invasiveness and migration of NPC cells. MiR-572 was highly expressed in NPC tissues as well as NPC cells, and there was lower expression of PPP2R2C in NPC tissues compared with normal samples. MiR-572 could bind to the 3' UTR of PPP2R2C and decrease its expression. Over-expressed miR-572 and decreased PPP2R2C expression could both inhibit proliferation and invasion and induce apoptosis of NPC cells. Thus, miR-572 promotes the proliferation and invasion of NPC by directly down-regulating PPP2R2C.

High-level expression and characterization of bioactive human truncated variant of hepatocyte growth factor in Escherichia coli.

Hepatocyte growth factor (HGF) is an effective anti-fibrotic factor because of its bioactivity in inhibiting fibrosis-related proteins in the development of hepatic fibrosis. However, high-level production of bioactive mature form HGF is difficult because of its complex structure. Here, we report a non-fusion protein expression system to obtain truncated variant of N-terminal hairpin and first kringle domains of HGF (tvNK1) in Escherichia coli to determine its anti-fibrotic effects on hepatic stellate cells (HSCs). Under the selected conditions of cultivation and isopropyl-ß-D-1-thiogalactopyranoside induction, the expression level of tvNK1 accounted for approximately 65 % of the total cellular protein and 50 % of fusion protein in the supernatant of whole cell lysates. The recombinant protein could be purified in one step with Ni(2+)-affinity chromatograph. Finally, about 65 mg recombinant tvNK1 was obtained from 1 l fermentation culture with no <95 % purity. In vitro, the final purified tvNK1 was shown to inhibit the proliferation of HSCs and decrease the mRNA and protein expression levels of fibrosis-related COL1A1 and α-smooth muscle actin genes.