OBJECTIVE: To investigate expression status and clinical relevance of SERPINE1 mRNA in esophageal squamous cell carcinoma (ESCC) tissues. METHODS: Expressions of SERPINE1 mRNA were determined using tissue microarrays and RNAscope in situ hybridizations in surgically-resected tumor tissues and adjacent normal epithelia from 236 ESCC patients. Relationships between the mRNA expression and clinicopathological characteristics as well as prognosis were analyzed. RESULTS: Increased expression of SERPINE1 mRNA was observed in 23.7% (56/236) of the tumor tissues. However,there was no expression of SERPINE1 mRNA in normal tissues which were adjacent to the carcinomas. The increased expressions were positively correlated with genders,histological grades,T stages and clinical stages (r=7.939,6.618,10.939,6.671,respectively;each P<0.05). Kaplan-Meier survival curves show that patients with high expressions of SERPINE1 mRNA had shorter survival times than those with low expressions (P<0.05). Cox multiple regression analyses indicate that SERPINE1 mRNA expression was an independent prognosis factor in ESCC. CONCLUSION: SERPINE1 mRNA was expressed highly in ESCC tissues. Our data indicate that the expression may serve as a molecular biomarker for predicting poor prognosis of ESCC patients.

OBJECTIVE: To investigate clinical significance,prognostic value and immune cell infiltration from connexin 43 (CX43) gene expression in gliomas. METHODS: Based on collected mRNA data and related clinical information among glioma patients in the Cancer Genome Atlas,the R language was used to analyze differential expressions of the CX43 gene in 592 patients. The Kaplan-Meier survival analysis method was used to evaluate its prognostic value. The Gene set enrichment analysis (GSEA) was used to explore potential mechanisms of CX43 involvement in glioma. Relationships between CX43 mRNA expression and immune cells infiltration into glioma were investigated using the CIBERSORTx algorithm. Finally,34 post-operative specimens were used for immunohistochemical analyses. RESULTS: Expression levels of CX43 mRNA were significantly increased in the wild-type isocitrate dehydrogenase and the chromosome 1p/19q non-co-deletion groups (P<0.01). Prognosis of glioma patients with high expression of CX43 were poor (P<0.01). GSEA results indicate that the high CX43 mRNA expression group showed enrichment in a 9 hallmark gene set. CIBERSORTx immune infiltration analyses show that monocytes,macrophages,M2 phenotypes,and activated dendritic cells were significantly increased in the high CX43 mRNA expression group (P<0.05). Results from the immunohistochemistry analyses indicate that high expressions of the CX43 protein were related to the absence of 1p/19q co-deletion and the increased CD163-positive M2-like macrophages. CONCLUSION: Our results indicate that high expression of CX43 was related to the pathological classification of glioma 1p/19q without co-deletion, to poor prognostic for glioma and to inducing macrophages to polarize to the M2 phenotype.

OBJECTIVE: This study was designed to investigate effects from 90-day exposure of yttrium nitrate on neurobehavioral functions in female rats. METHODS: Female Sprague-Dawley rats (PND 21),weighing 45-55 g each,were randomly divided into 4 groups of 15 rats per group:control (ddH₂O exposure) and exposed groups[10,40,160 mg/(kg·d) of Y(NO₃)₃]. Rats in the exposed group were administered orally with Y(NO₃)₃ for 90 days. Then,all exposed and control rats were evaluated using Open field,Elevated plus maze,Rotarod and Morris water maze tests. After the tests,5 rats per group were perfused in situ. Their brains were removed and fixed for histopathological analyses. The other 10 rats per group were sacrificed. Their brains were removed and the levels of glutamate (Glu) in cerebral cortex and hippocampus were determined using UV-Vis. Protein expression levels of NMDARs in hippocampus were detected by Western blot. RESULTS: After 90 days of Y(NO₃)₃ exposure,the 160 mg/(kg·d) group showed significantly increased average time on the rotarod compared to the control group. In the Morris water maze test,the escape latencies among rats in the 40 and 160 mg/(kg·d) Y(NO₃)₃ groups were significantly reduced compared to that in the control group at the 4th day of navigation test. In the space exploration test of Morris water maze test,the times crossing platform and the times required for swimming in target quadrant were significantly increased in the 40 mg/(kg·d) Y(NO₃)₃ group. The times crossing platform,entering the target quadrant and swimming time in the target quadrant were significantly increased in the 160 mg/(kg·d) Y(NO₃)₃ group. The escape latency of NE,SW and SE quadrants in the 160 mg/(kg·d) Y(NO₃)₃ group was significantly delayed compared to that in NW quadrant. Glu level in hippocampus in the 160 mg/(kg·d) Y(NO₃)₃ group was significantly decreased. Expressions of NMDAR2A in hippocampus were significantly decreased in the 40 and 160 mg/(kg·d) Y(NO₃)₃ groups. Expressions of NMDAR1 in the hippocampus were significantly decreased in the 160 mg/(kg·d) Y(NO₃)₃ group. CONCLUSION: Subchronic (90-day) exposure to Y(NO₃)₃ affected the spatial learning and memory of female rats,possibly by decreasing Glu in the extracellular of hippocampal neurons and by antagonizing the activation of NMDARs.

OBJECTIVE: To explore the role of estrogen receptor α (ESRRA) in development of lung cancer using bioinformatics methodology. METHODS: The UALCAN online platform was used to analyze gene expression levels in lung cancer tissues,e.g.,adenocarcinomas and squamous cell carcinomas (LUAD and LUSC) and adjacent normal tissues. The Kaplan-Meier Plotter database was used to perform patients' survival analyses. The GEPIA2 online analysis platform was used to obtain the top 50 genes which had similar expression patterns to that of ESRRA. The protein-protein interaction (PPI) network was constructed using the GeneMANIA online analysis platform. GO analysis and KEGG pathway enrichment were performed for gene clusters in the David database. The TIMER2.0 online analysis platform was used to analyze correlations between ESRRA and gene expression levels which were enriched in oxidative phosphorylation pathways in lung cancer tissues. Expression levels of ESRRA were analyzed using Western blot. RESULTS: Transcription levels of ESRRA in LUAD and LUSC tissues were significantly higher than those in normal lung tissues (P<0.01),and the higher levels of ESRRA were associated with poorer lung cancer prognosis (P<0.01). Expression levels of the top 50 genes which were similar to that of ESRRA were mainly enriched in the oxidative phosphorylation pathways in lung cancer tissues. These genes were mainly:ATP5B,ATP5G3,COX5A,COX8A,SDHD,UQCRC1 and UQCRC2. Moreover,expressions of ATP5B,COX8A and UQCRC1 were positively correlated with that of ESRRA (P<0.05),and that of ATP5B,ATP5G3,COX5A with poorer prognosis (P<0.01). On the other hand, reduced expression levels of SDHD indicate poorer prognosis (P<0.01). Western blotting results indicate that the relative expression of ESRRA in NCI-H1975 cells was higher than that in 16HBE cells (P<0.01),and the relative expression of ESRRA in SW900 cells was higher than that in 16HBE cells (P<0.01). CONCLUSION: ESRRA-mediated changes in the expressions of ATP5B and COX8A in the oxidative phosphorylation pathways which were mainly involved with energy metabolism disorder. Our data suggest that ESRRA contributes to the development and prognosis of lung cancers.

OBJECTIVE: To investigate immunotoxicity of triphenyl phosphate (TPHP) on mouse RAW264.7 in vitro. METHODS: Raw264.7 cells were treated with different concentrations of TPHP (0,12.5,25,50,100,200,400 μmol/L) for 24,48 or 72 h. CCK-8 colorimetry and lactate dehydrogenase (LDH) release assays were used to determine cell activities. Flow cytometry was used to detect phagocytosis of macrophages to fluorescent microspheres. Western blot was used to detect changes in expression levels of Toll-like receptor 4 (TLR4),serine/threonine kinase (Akt) and mammalian target of rapamycin (mTOR). RESULTS: Compared with the control group,survival rates of RAW264.7 cells were reduced together with increase of TPHP exposure concentrations and times,therefore, significant dose-(r=0.960,P<0.05) and time-dependent (r=0.980,P<0.05) toxicity. Levels of LDH in cell culture medium increased significantly when the exposure concentrations were 50-400 μmol/L (P<0.05). When exposed to 25 and 50 μmol/L of TPHP,percentages of FITC-A⁺ cells were significantly increased, indicating significant increases of phagocytosis of fluorescent microspheres (P<0.05). Western blot results show that TPHP increased expression of TLR4 protein and triggered phosphorylation of Akt and mTOR (P<0.05 or P<0.01). CONCLUSION: TPHP exhibitedsignificant cytotoxicity to Raw264.7 cells,leading to enhanced phagocytosis of macrophages,and increased expressions of TLR4,Akt and mTOR.

OBJECTIVE: To evaluate expressions of protein tyrosine phosphatase receptor J (PTPRJ) in breast cancer tissues and their roles in clinical process. METHODS: The immunohistochemical SP method was used to detect expressions of PTPRJ in normal breast and breast cancer tissues. Then,relationships between PTPRJ expressions,and clinicopathological parameters and survival prognosis of breast cancer patients were analyzed. RESULTS: Results from immunohistochemical staining show that expressions of PTPRJ in normal breast tissues were significantly higher than that in breast cancer tissues (P=0.003). PTPRJ expressions were significantly correlated with lymph node metastasis (P=0.014),TNM stages (P=0.003) and ER status (P=0.048). Kaplan-Meier analyses reveal that breast cancer patients with low PTPRJ expressions in tumors had significantly worse overall survival rates than those with higher PTPRJ expressions (P=0.002). Multivariate analyses show that TNM stages and PTPRJ expressions were independent survival predictors. CONCLUSION: Our data indicates that expressions of PTPRJ were reduced in breast cancer tissues and were closely related to tumor progression. Therefore,PTPRJ expression can possibly be used as a valuable marker for prognosis in patients with breast cancers.

OBJECTIVE: To investigate effects of liraglutide on cognitive function and phosphorylation of Tau protein in a streptozotocin (STZ)-induced Alzheimer's disease (AD) in rats. METHODS: 24 male Wistar rats were randomly divided into blank control,sham operation,model and treatment groups,with 6 rats per group. For an AD rat model,each rat received a single injection of STZ (3 mg/kg) into the right intracerebroventricular region. After 14 days,rats in the treated group received subcutaneous injections of liraglutide for 4 weeks. Cognitive functions were detected using the Morris water maze test. Expressions of phosphorylation of Tau protein Ser396 were detected using immunohistochemistry (IHC) and Western blot methods. RESULTS: Results from the Morris water maze test show that both the escape latency and the number of times of crossing the platform for rats from the sham operation group were not significantly different from the blank control group (P>0.05). From the same test,results from the model rats were markedly increased over the sham operation group (P<0.05), that from the treated group were markedly decreased compared to the model group (P<0.05). Results from the IHC and Western blot analyses show that phosphorylations of the Tau protein Ser396 were increased significantly in the model rats (P<0.05) but decreased after liraglutide treatments (P<0.05). CONCLUSION: Our data show that liraglutide improved the STZ-induced damage of spatial learning and memory abilities and reduced expression of phosphorylation of Tau protein Ser396 in the hippocampi of rats.

OBJECTIVE: To investigate correlations between glutathione peroxidase (GSH-PX) in serum and clinicopathological indicators,curative effect of combined radio-with chemotherapy and prognosis of advanced non-small cell lung cancers (NSCLC). METHODS: In 2018,174 newly diagnosed advanced NSCLC patients (before anticancer therapy) from the Department of Oncology,Affiliated Hospital of Hebei University of Engineering and 80 healthy controls were recruited for our investigation. Expressions of GPX3 in different tumors were analyzed via the data of on-line. Activities of GSH-PX in serum from peripheral blood samples were detected via 5',5-dithio-bis-nitrobenzoic acid (DTNB). Activities of GSH-PX were compared between the advanced NSCLC and the control groups. In addition,relationships between activities of GSH-PX and clinicopathological indicators,curative effect of combined radio-with chemotherapy and prognosis of advanced non-small cell lung cancer were analyzed. RESULTS: Data from the gene expression profiling interactive analysis (GEPIA) show that expressions of GPX3 in 33 kinds of tumors (lung squamous cell carcinoma and lung adenocarcinoma) were significantly lower than that in the controls (P<0.05). Results from DTNB show that GSH-PX activities in serums of patients were significantly lower than that in the controls (P<0.05) and were irrelevant to the age,gender and pathological patterns of patients (P>0.05). However GSH-PX activities were negatively related to smoking histories and tumor sizes (r=-0.295、0.228,all P<0.05),and positively related to KPS scores and pathological grades (r=0.295、0.228,P all <0.05). There was no difference in GSH-PX activities between patients with effective and failed treatments (P>0.05). Overall survivals in the high activities group was higher than those in the low activities group (P<0.05),and they were positively related (r=0.238,P=0.000). CONCLUSION: GSH-PX activities in serum of patients with advanced NSCLC were significantly reduced,and the activities were related to smoking histories,KPS scores,tumor sizes,pathological grades and overall survivals. Therefore,GSH-PX activities were involved in the development and prognosis of NSCLC.

OBJECTIVE: To study effects of smoke exposure on gene expression in hearts of rats using gene chip technology. METHODS: Six male SD rats were divided into two groups:control (n=3) and smoke exposure (n=3). The exposure group were exposed to mainstream smoke for 30 days. Rat hearts were removed and differentially expressed genes were screened using gene chip technology. Differentially-expressed genes were analyzed using gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) databases. RESULTS: Compared with the control group,86 genes with differential expression of more than 2 times were screened out in the smoke exposure group,among which 60 genes were up-regulated and 26 genes were down-regulated. Differentially expressed genes were enriched in 292 biological processes:72 cellular components,96 molecular functions and 61 signaling pathways. These processes were involved in mitogen-activated protein kinase binding,alpha-beta T cell receptor complex,positive regulation of cellular extravasation,and tyrosine metabolism. CONCLUSION: After exposure to smoke for 30 days,differentially expressed genes and signaling pathways in the hearts of rats were mainly involved in molecular binding,cell cycle control and signal transduction,etc.

OBJECTIVE: To investigate clinical features, laboratory findings and survival in myeloid leukemia patients who had +1,der (1;7)(p10;q10) or -7/7q- abnormal karyotypes,and to guide their clinical diagnosis,treatment and prognosis. METHODS: From January 2013 to November 2020,clinical data of 21 patients with the mentioned karyotype abnormalities,including myelodysplastic syndrome (MDS),acute myeloid leukemia (AML) and myeloproliferative neoplasm (MPN),were analyzed. Bone marrow fluid samples were collected and used to detect 38 disease-related gene mutations using a next generation sequencing method and collected data were evaluated to achieve our objectives. RESULTS: Among the 21 patients,those with der(1;7) were predominantly male (P=0.024),had significantly lower platelet counts (P=0.036) and higher CRP (P=0.029) than those with -7/7q-. Immunophenotypes of the two groups were similar (χ²=0.078,P=1.000). There were 37 mutations in 13 genes among 15 patients (71.4%). The mutation frequencies of RUNX1 (21.4%),IDH1/IDH2 (21.4%) and JAK2 (21.4%) were higher in patients with der(1;7),and SETBP1 (17.4%),TET2 (17.4%),ASXL1 (13%),U2AF1 (13%) and DNMT3A (13%) were higher in patients with 7/7q- than the comparison groups. There was significant differences in gene mutation compositions between the two groups (P=0.012),and significant correlations between gene mutations and karyotype abnormalities (r_s=0.522,P=0.001). The der(1;7) patients had significantly shorter overall and median survival durations than the -7/7q- patients (P=0.045,HR=2.844). In the der(1;7) patients,the median survival times of those with the RUNX1 mutation were shorter than those with the wild type (P=0.435). The median survival and total survival times for those with the ASXL1 mutation were significantly shorter than those with the wild type among the 7/7q- patients (P=0.018).CONCLUSION: Our data indicate that myeloid leukemia with the two abnormal karyotypes:der(1;7) or -7/7q-,were independent disease subtypeswith significant differences in laboratory findings,gene mutations and clinical outcomes.

OBJECTIVE: To investigate inhibitory effects and the mechanisms of farnesoid X receptor (FXR) on proliferation of a cervical cancer cell line,CaSki. METHODS: FXR was activated by the FXR agonist GW4064 and DMSO was set as the control group at the same time. After treatment with 10 μmol/L GW4064 for 24 and 48 h,the mRNA and protein levels of FXR were detected by fluorescence quantitative PCR and Western blot,respectively. Proliferations of CaSki were detected by using methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay after FXR activation. Cell apoptosis of CaSki was detected by flow cytometry. Western blot was used to detect protein levels of p53. RESULTS: Compared with the control group,expression levels of FXR mRNA and protein in the GW4064 treatment group were increased,cell proliferation rates were decreased,apoptosis rates were increased,and expression levels of p53 protein were increased (all P<0.05). CONCLUSION: Activation of FXR inhibited proliferation of CaSki through promoting cell apoptosis which might be related to increased expression of p53.

OBJECTIVE: To investigate sensitivities of two methods and four time points for detecting muscle implantation. Consequently,the rabbit tissue response grading method and semi quantitative score evaluation method were used to detect tissue stimulation responses to seven kinds of medical materials after short-term and long-term muscle implantation. METHODS: The left and right sides of 84 New Zealand white rabbits were compared in this investigation. The same test and control samples were implanted into the left and right paraspinal muscles at 4 implantation points with an interval of 2.5 cm. On the 15th,30th,60th and 90th day after implantation,3 rabbits were killed respectively,and the 1.0-1.5 cm³ of back muscle tissues which contained the implanted samples were removed. The numbers of inflammatory cells were counted microscopically. Proliferative reaction indexes were graded and semi quantitative scorings were performed according to the GB/T 16886 standard. The gradings or stimulation degrees were determined according to the difference between the total average scores from the left and right sides. RESULTS: The samples which showed mild irritation by tissue response grading method were No.3,No.5 (60 days after implantation) and No.3 (90 days after implantation). The samples which showed mild irritation by the semi quantitative score evaluation method were No.5 (30 days after implantation),No.3,No.4,No.6 (60 days after implantation),No.3 and No.5 (90 days after implantation). The sensitivity of the two methods and time points to observe inflammatory reactions and tissue reactions were different. CONCLUSION: The semi-quantitative score evaluation method was better than the tissue reaction grading method. In addition,multiple implantation time points were better than one time point and the detection sensitivities for sample implantation were better after 60 d.

OBJECTIVE: To develope a solution which could be used for preserving DNA samples in fecal samples from individuals who participated in early colorectal cancer screening. METHODS: An original formula was modified for this investigation. Fecal samples were stored at room temperature for 3 and 7 days,and freezing and thawing at -80℃ within the 7 days. Then,agarose gel electrophoresis (the qualitative method) and real-time fluorescent quantitative PCR (qPCR,the semi-quantitative method) were used for detecting presence of the human housekeeping β-globin gene. Usefulness of our modified preservation solution wascompared to similar international preservation solutions using 501 of the fecal samples. RESULTS: The agarose gel electrophoresis band did not change significantly after the fecal samples were stored at room temperature for 3 and 7 days. The qPCR results as measured using the ΔC_t value were less than or equal to 0.2. After freezings and thawings,the agarose gel electrophoresis results remained the same and ΔC_t value for the qPCR results was less than 1. Compared with the international preservation solutions,our solution showed more clear bands and less changes of the ΔC_t value. Using our solution,effectiveness in preserving fecal samples for DNA analysis was >70% compared to <40% if no preservation solutions were used. CONCLUSION: Our data indicate that our preservation solution was both economical and effective in preserving fecal samples after cold-chain transportation and in inhibiting proliferation of microorganisms. Our solution is therefore useful for use in large-scale colorectal cancer screening.