Objective:To evaluate the value of pretreatment 18F-fluorodeoxyglucose (FDG) PET/CT-based heterogeneity for early prediction of targeted therapy outcome in patients with human epidermal growth factor receptor 2 (HER2) positive metastatic breast cancer. Methods:From May 2012 to April 2018, 29 patients (all females, median age: 52 (32-69) years) who had HER2 positive metastatic breast cancer and underwent pretreatment 18F-FDG PET/CT in Fudan University Shanghai Cancer Center were retrospectively enrolled. All patients received trastuzumab as first-line treatment and were followed up for 6-87 (median time: 35) months. The relations between clinicopathologic parameters or PET/CT-based parameters and progression-free survival (PFS)/overall survival (OS) were analyzed with Cox univariate analysis. The parameters with P≤0.01 were further analyzed with Cox multivariate analysis. Optimal cut-off values were determined by time-dependent receiver operating characteristic (ROC) curve analysis. The survival analyses were estimated by Kaplan-Meier method and log-rank test. Results:The median OS of the 29 patients was 30 (6-83) months, and the median PFS was 10 (2-29) months. The PET/CT-based heterogeneity index(HI), including the maximum standardized uptake value (SUV max) ratio (SUV max-R; hazard ratio ( HR)=8.6, 95% CI: 2.7-27.8, P<0.001), the mean standardized uptake value (SUV mean)-2.5 (the cut-off value of standardized uptake value (SUV)=2.5) ratio (SUV mean-2.5-R; HR=2.6, 95% CI: 1.2-5.9, P=0.020), the metabolic tumor volume(MTV)-2.5 ratio(MTV-2.5-R; HR=2.4, 95% CI: 1.1-5.2, P=0.030), and the total lesion glycolysis(TLG)-2.5 ratio(TLG-2.5-R; HR=3.2, 95% CI: 1.4-7.4, P=0.008) of the lesion with the highest SUV max to that with the lowest SUV max, were significantly associated with PFS. None of the parameters was significantly associated with OS (all P>0.05). Multivariate analysis showed that the SUV max-R was the only independent predictor for PFS ( HR=6.8, 95% CI: 1.8-26.1, P<0.01). Area under the ROC curve for SUV max-R was 0.747. With a cut-off value of 1.8, SUV max-R could effectively distinguish the benefit from non-benefit population treated with trastuzumab (15.0 vs 7.0 months; χ2=18.68, P<0.01). Conclusion:Pretreatment 18F-FDG PET/CT-based HI has potential value for early prediction of first-line trastuzumab treatment outcome in patients with HER2 positive metastatic breast cancer.

Objective To study the effect of osgentide (OST) on proliferation of mouse preosteoblast MC3T3-E1 under simulated microgravity ( SMG ) .Methods Under normal conditions , cell proliferation was evaluated by MTT assay to screen an OST compound of an effective concentration after MC 3T3-E1 cells were treated with series OSTs .Furthermore, cell proliferation and cell cycle distribution of MC 3T3-E1 cells were analyzed after treatment with 1 nmol/L OST5 by MTT assay and by flow cytometry ( FCM) scanning under SMG .Results Under normal conditions , 1 nmol/L OST5 was able to significantly promote the proliferation of MC3T3-E1 cells (P<0.01).Under SMG, proliferation of MC3T3-E1 cells was significantly inhibited and more cells entered G 1 than under normal conditions (CN).The proportion of S phase of MC3T3-E1 cells after treatment with 1 nmol/L OST5 ( OST-SMG) for 3 d was higher than that of untreated MC 3T3-E1 cells under SMG,suggesting that OST5 could promote DNA synthesis ( P<0.05 ) .Conclusion OST5 facilitates the proliferation of MC3T3-E1 cells under SMG, which provides a basis for the use of OST5 in the prevention and treatment of bone loss relat-ed to microgravity .

Objective To investigate the effects of simulated microgravity by rotary cell culture system (RCCS) on expression profiles of miRNA in mouse fibroblasts L929 cell line.Methods L929 cells were cultured in vitro and divided into simulated microgravity (SMG) group and normal gravity (NG) group according to the simple random method.Samples of two groups were collected on 7th day of culture and the total RNAs were extracted,labeled,and hybridized in sequence.Feature Extraction Software was used to collect the array images and get raw data,which were analyzed by Genespring Software.Differentially expressed miRNAs were identified and then validated by qRT-PCR.Target genes of differentially expressed miRNAs were predicted by the databases of Targetscan and microRNAorg,and the intersections of databases were identified as potential regulatory target genes.Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) analysis were applied to determine the roles of these target genes.Relevant biological functions and/or signaling pathways of the regulated genes especially related with wound healing process were categorized based on their enrichments.Then integration predictions of the miRNA and mRNA expression profiles had been proposed to refine the functional miRNA-mRNA relationships.The miRNA-mRNA functional network and miRNA-mRNA control network were constructed.Results Four miRNA genes were up-regulated significantly including mmumiR-669j,-122-5p,-30a-3p,-6516-3p,among which mmu-miR-669j was up-regulated at 52.84 folds with the greatest significance (P ＜ 0.05).Seventeen miRNA genes were down-regulated significantly including mmu-miR-21a-3p,-miR-28a-5p,-218-5p,-210-3p,-miR-19a-3p,-miR-31-3p,and-miR-19b-3p,among which mmu-miR-28a-5p was down-regulated at 15.47 folds with the greatest significance (P ＜ 0.05).The qRT-PCR showed a high concordance with the microarray results (P ＜ 0.05).Target gene prediction and functional enrichment analysis suggested that a variety of biological processes and signaling pathways involved in wound repair were significantly enriched (P ＜ 0.05).Function network and regulation network of miRNA-mRNA covered all the differentially expressed miRNAs,which suggested that miR-21 a-3p and predicted target gene Smad3 might play an important role in wound healing under microgravity.Conclusions Simulated microgravity by RCCS can significantly affect the expression of stress-related miRNAs in mouse fibroblasts L929.The miRNA target gene prediction and functional enrichment analysis based on gene chip technology may provide theoretical basis for illustrating the mechanism and management of weightlessness stress injury.

Objective To investigate the effect of imatinib on the growth of A549 non-small cell lung cancer transplanted tumors and the expression of PDGFB and PDGFRβ proteins in tumor tissues and stroma in nude mice and to explore the underlying tumor suppression mechanism. Methods A transplantation tumor model of A549 non-small cell lung cancer was established in nude mice. The mice were then randomly divided into four groups: control group (0.9%NaCl), low-dose imatinib group (50 mg/(kg·d)), medium-dose imatinib group (100 mg/(kg·d)), and high-dose imatinib group (200 mg/(kg·d)). The effect of different concentrations of imatinib administered by continuous gavage on tumor growth was observed for 28 days. HE staining was performed to observe the pathological changes of tumor tissues. The expression of PDGF/PDGFR pathway-related proteins and the phosphorylation levels of AKT and ERK1/2 proteins in tumor tissues were detected by Western blot analysis. Double immunofluorescence staining was used to detect the expression of PDGFB and PDGFRβ proteins in the tumor stroma. Results Imatinib inhibited the growth of A549 non-small cell lung cancer cells in nude mice, suppressed the expression of PDGFB in tumor tissues, and decreased the phosphorylation levels of PDGFRβ, AKT, and ERK1/2. The expression of PDGFB and PDGFRβ in tumor stromal fibroblasts of the administered group was significantly lower than that of the control group. Conclusion Imatinib exhibits a pronounced inhibitory effect on A549 xenografts of nude mice with non-small cell lung cancer, and its antitumor mechanism may involve the downregulation of PDGFB and PDGFRβ expression in tumor stromal fibroblasts.

BACKGROUND:Astrocytes play an important role in the pathology of central nervous system diseases.The phenotypic and functional changes in astrocytes suggest that it may be an effective therapeutic target for central nervous system diseases.Our previous studies have confirmed that astragaloside can inhibit the lipopolysaccharide-induced astrocyte inflammatory response.Whether astragaloside can regulate the phenotype and function of astrocytes through Notch-1 and its downstream signaling pathway remains unclear. OBJECTIVE:To explore the effect of astragaloside on astrocyte activation and inflammatory response induced by inflammation and its possible mechanism. METHODS:Cerebral cortex astrocytes derived from neonatal C57BL/6 mouse were cultured in vitro.CCK-8 assay was used to determine the optimum concentration of astragaloside and Notch active inhibitor DAPT.The astrocytes were divided into five groups:PBS group,lipopolysaccharide group,lipopolysaccharide + astragaloside group,lipopolysaccharide + DAPT group and lipopolysaccharide + DAPT + astragaloside group.The secretion level of inflammatory factors was detected by ELISA,and the level of nitric oxide was detected by Griess method.The astrocytes and splenic mononuclear cells were co-cultured in Transwell chamber to observe the migration of CD4T cells.The expression of astrocyte activation marker GFAP,A1 marker C3 and A2 marker S100A10 as well as Notch 1 and Jag-1 was detected by immunofluorescence staining.The expressions of CFB,C3,S100A10,PTX3,Notch-1,Jag-1,and Hes were detected by western blot assay. RESULTS AND CONCLUSION:(1)According to the results of CCK8 assay,the final concentration of astragaloside was selected as 25 μmol/L and the final concentration of DAPT was 50 μmol/L for follow-up experiments.(2)Compared with PBS group,interleukin-6,interleukin-12 and nitric oxide secretion levels in the lipopolysaccharide group were significantly increased(P<0.05,P<0.05,P<0.01).Compared with the lipopolysaccharide group,interleukin-6(all P<0.05),interleukin-12(P>0.05,P<0.05,P<0.05)and nitric oxide(P<0.05,P<0.01,P<0.01)secretion significantly reduced in the lipopolysaccharide + astragaloside group,lipopolysaccharide +DAPT group,lipopolysaccharide + DAPT + astragaloside group.(3)Compared with the PBS group,the expression of GFAP that is the marker of activated astrocytes and the migration of CD4 T cells were significantly increased in the lipopolysaccharide group(P<0.01).Compared with the lipopolysaccharide group,astrocyte activation was significantly inhibited and CD4 T cell migration was significantly reduced in the lipopolysaccharide + astragaloside,lipopolysaccharide +DAPT,lipopolysaccharide + DAPT + astragaloside group(P<0.05,P<0.05,P<0.01).(4)Compared with the PBS group,the expressions of A1 markers C3 and CFB in the lipopolysaccharide group were increased(P<0.01,P<0.05),and the expressions of A2 markers S100A10 and PTX3 were decreased(P<0.01,P<0.05).Compared with the lipopolysaccharide group,C3(all P<0.01)and CFB(both P<0.05)were significantly reduced and S100A10(all P<0.01)and PTX3(P<0.05,P<0.05 and P>0.05)were increased in the lipopolysaccharide + astragaloside,lipopolysaccharide +DAPT,lipopolysaccharide + DAPT + astragaloside group.(5)Compared with the PBS group,the expressions of Jag-1,Notch-1 and Hes in the lipopolysaccharide group were significantly increased(all P<0.01).Compared with the lipopolysaccharide group,the expressions of Jag-1(all P<0.01),Notch-1(all P<0.01)and Hes(P<0.05,P<0.01 and P<0.01)were significantly reduced in the lipopolysaccharide + astragaloside,lipopolysaccharide +DAPT,lipopolysaccharide + DAPT + astragaloside group.(6)The results indicate that astragaloside can promote the transformation of astrocytes from A1 to A2 by regulating Notch-1 signaling pathway,reduce the secretion of inflammatory factors and the migration of CD4 T cells,and thus inhibit astrocyte activation and inflammatory response.