OBJECTIVE: To evaluate the effects of SOX30 gene knockout on testicular tissue structure and hormone secretion in male mice,and to explore the role of SOX30 gene in male reproductive system. METHODS: The SOX30 gene knockout mouse model was constructed by the gene recombination method and genotypes were identified by the PCR method. The male mice were divided into wild type (WW),heterozygous (KW) and homozygous (KK) groups according to the results from genotype identification (2.5 months old). Testicular volume and testicular organ index were calculated. After H&E staining,structures of the testes and epididymes of each mouse were determined,and diameters of the Seminiferous tubule were measured. Levels of the testosteron (T) and inhibin B (INH-B) hormones in homogenates of testicular tissues were detected by ELISA. Expressions of SOX9 and 3β-HSD in testicular tissues were determined by using the immunofluorescence technique. RESULTS: Compared with the WW and KW types,the testicular organ index of KK mice was significantly decreased (P < 0.05). There were no pathological changes in the testes and epididymes of KW and WW mice. On the contrary,testes in KK mice had disordered spermatogenic cells,giant cells,vacuolar degeneration and interstitial hyperplasia. No matured sperm was found in the epididymis of KK mice. The lumen diameters of seminiferous tubules in the KK mice were reduced compared with the WW mice (P < 0.05). In addition,the T and INH-B levels in homogenates from testicular tissues were reduced significantly in the KK compared with the WW and KW mice (P < 0.05). There was no significant difference in the number of Sertoli cells expressing SOX9 between WW and KK mice,but the number of leydig cells expressing 3β-HSD in KK mice was significantly more than that in WW mice (P < 0.05). CONCLUSION: Our results suggest that the knockout of SOX30 induced pathological damage in testes of mice via inhibiting the synthesis of T and INH-B which played important roles in maintaining normal functions of testes.

OBJECTIVE: To explore interactions between nucleus receptor RXRs and other nuclear receptors in BDE-47-induced cytotoxicity in human neuroblastoma SK-N-SH cells. METHODS: RXRα-knockout cell lines and RXRα high-expression cell lines from human neuroblastoma SK-N-SH cells (named RXR/KO-SK-N-SH and RXR/OE-SK-N-SH cells,respectively) were constructed. The three cell lines were exposed to different concentrations of BDE-47 (1,5,10,20,50,100,150,200 mol/L) for 12,24,48,72 h,and their proliferation rates were determined by the CCK-8 method. mRNA and protein levels of RXR,TR,TR,PPAR and PPAR were measured by qPCR and Western blot,respectively. RESULTS: Proliferation rates of three cell lines which were exposed to BDE-47 for 12,24,48,72 h were significantly different from each other,(P < 0.05). The IC₅₀ of BDE-47 to RXR/KO-SK-N-SH was 1/2 to SK-N-SH and RXR/OE-SK-N-SH. After treatment with BDE-47,the wild-type and RXR/OE-SK-N-SH cell lines showed significant increase of RXRα mRNA and protein expression levels. Their protein expression levels of TRα and PPARα showed the same changes. However,the knockout SK-N-SH cells showed lower expression level in RXRα. The expression levels of TRβ and PPARγ were not dramatically changed in all three SK-N-SH cell lines after the BDE-47 treatment. CONCLUSION: RXRα played a critical role in improving cell viability. After treatment with BDE-47,RXRα mediated the expression of TRα and PPARα in the three SK-N-SH cell lines,but not the expression of TRβ and PPARγ. These data demonstrate that toxicity of BDE-47 on SK-N-SH cells was medicated by activation of RXRα which enhanced TRα and PPARα expression.

OBJECTIVE: The aim of this study was to investigate associations between expression of CD10 protein and clinicopathological characteristics in colorectal cancer patients. METHODS: 78 cases of colorectal cancer tissue samples and 22 cases of neighboring noncancer colorectal tissue samples were collected from colorectal cancer patients. In addition,expression of CD10 protein in tissue samples was determined using rapid immunohistochemical EliVision method. Relationships between the expression intensity and clinicopathological characteristics of colorectal cancer were analyzed by the Chi Square test. RESULTS: There was no expression of CD10 protein in 22 normal colorectal tissue samples. The positive rate of CD10 expression was significantly different in colorectal cancer tissue samples and in neighboring noncancer colorectal tissue samples:46.2%(36/78) and 0,(P < 0.01). The expression intensity of CD10 protein in cancer tissues was not clearly related to gender,age,tumor location,differentiation degree and depth of invasion (P > 0.05) but was higher in the more advanced stages than in the early stages (P=0.033),and was also associated with lymph node metastasis (P=0.023) and vessel invasion (P=0.004). On the other hand,the data show no significant correlation between the expression ratio of CD10 protein and clinicopathological characteristics of colorectal cancer (P > 0.05). CONCLUSION: CD10 protein was highly expressed in colorectal cancer tissues and the expression intensity was associated with clinical stage,lymph node metastasis and vessel invasion,which suggests that CD10 protein may play roles in promoting the development,invasion and metastasis of colorectal cancer.

OBJECTIVE: To investigate relationships between clinicopathological characteristics and curative effect of combined radio-with chemo-therapy for advanced non-small cell lung cancer (NSCLC) with superoxide dismutase (SOD). METHODS: 174 newly diagnosed advanced NSCLC patients in the Department of Oncology,Affiliated Hospital of Hebei University of Engineering were selected in 2018 as the research subjects,and 80 healthy subjects were selected as controls. Peripheral blood and serum samples were collected from patients before they had any therapy. Expression of SOD in different tumors was analyzed via gene expression profiling interactive analysis (GEPIA) and activities of SOD in serum were detected via WST-1. Then,relationships between their expressions,clinicopathological chara cteristics and curative effect of therapy were analyzed. RESULTS: On-line analyses via GEPIA show that the expression of SOD in 19 kinds of tumors was abnormal among 33 kinds of tumors. Expression of SOD in lung squamous cell carcinoma (LUSC) and lung adenocarcinoma (LUAD) were significantly lower than that in the control group (P < 0.05). The results of WST-1 show that SOD activity in serum of patients was lower than that in the control group (P < 0.05),and there was no difference in serum SOD activity between different ages,genders and pathological patterns of patients (P > 0.05). However,there was significant difference in serum SOD activities between smoking history and pathological grades (P < 0.05). SOD activities were significantly lower in smoking patients compared with non-smoking patients (P < 0.05);and in patients with poor differentiating tumors compared with patients having better differentiating tumors (P < 0.05). SOD activities were positively related to the curative effect of combined therapies (r=0.554,P < 0.05),and it was higher in patients with effective treatment than patients with failed treatment (P < 0.05). Meanwhile,the overall survival in the high activity SOD group was higher than that in the low activity group (P < 0.05). CONCLUSION: SOD activities in serum of patients with advanced NSCLC were significantly decreased. The activities were affected by patients' smoking history,pathological grades,as well as to curative effects of combined therapies and OS. Therefore,SOD can be a potential indicator for evaluating curative effects and prognosis of NSCLC.

OBJECTIVE: To investigate effects of PM_2.5 on HBE cells using microRNA (miRNA) sequencing and bioinformatics methods. METHODS: HBE cells were treated with 50 μg/mL PM_2.5 suspension and untreated cells were used as control. Total RNA samples were extracted 24 hours later. The Small RNA sample pretreatment kit was used to construct a library and the illumina HiSeq^TM 2500/MiSeq sequencing platform was used for high-throughput sequencing. The differentially expressed miRNAs were obtained according to the screening criteria of adjusted P < 0.05,and the target genes of the differentially expressed miRNAs were predicted by miRanda and RNAhybrid software,and then GO and KEGG function enrichment analyses were performed. The STRING database and the Cytoscape software were used to visualize interactions between miRNAs and target genes and between target genes. RESULTS: Using high-throughput sequencing methods,a total of 1 137 miRNAs were detected before and after PM_2.5 exposure in HBE cells. Among them,27 miRNAs were differentially expressed,including 7 up-regulated and 20 down-regulated. The enrichment analyses using GO and KEGG show that these differential miRNAs were mainly involved in biological processes such as cell metabolism and enzyme activity regulation. They are mainly involved with metabolic,PI3K-Akt signaling,cancer,Ras signaling,MAPK signaling pathways,etc. related to cell proliferation,differentiation,apoptosis and cancer. By constructing an interaction network diagram,the core miRNAs with the number of TOP 11 interactions such as miR-371b-3p,miR-371a-5p,miR-27a-3p,miR-7-5p,miR-372-3p,and 3 core target genes (VEGFA,MAPK3,CCR5) were identified. CONCLUSION: 27 miRNAs were differentially expressed after HBE cells were exposed to PM_2.5. These miRNAs are involved with the PI3K-Akt signaling,Ras signaling,MAPK signaling pathways,etc. which are involved with the development of cancer. Among them,11 core miRNAs and 3 core genes were screened,our results provide the scientific basis for further study of the carcinogenic mechanism of PM_2.5.

OBJECTIVE: To investigate relationships between expression in the succinate receptor pathway and cardiomyocyte injury under a hypoxic environment. METHODS: Primary cardiomyocytes of suckling mice were cultured in vitro under different hypoxic environment. The siRNA interference technology was used to reduce expression of intracellular succinate receptor G protein-coupled receptor 91 (GPR91). The different cultures were randomly divided into the control (CTL),CTL+siGPR91 group,succinate group,succinate+siGPR91 group,hypoxic CTL group,hypoxic CTL+siGPR91 group,hypoxic succinate group and hypoxic Succinate+siGPR91 group. After 5 days of cultivation under 10% oxygen content,CCK-8 was used to detect the survival rate of cardiomyocytes. In addition,cellular ATP contents of primary cardiomyocyte hypoxia model were determined. Changes in expression of Akt and P-Akt in the PI3K/Akt protein pathway were determined using the Western blot. RESULTS: Compared with the CTL group,the survival rate and the content of ATP of myocardial cells were reduced,and expressions of BNP and CaMK Ⅱ were up-regulated in the hypoxic environment (P < 0.05). Compared with the CTL group,expression levels of GPR91 in the siRNA interference group were suppressed (P < 0.05). At the same time,myocardial cell survival rates and ATP contents were increased, BNP and CaMK Ⅱ protein expressions were down-regulated,and phosphorylation levels of PI3K and levels of downstream signaling molecule Akt were reduced (P < 0.05). CONCLUSION: Our data show that GPR91 was involved in the pathological process of myocardial ischemic injury by regulating the phosphorylation of PI3K/Akt pathway.

OBJECTIVE: To study the clinical significance of the expression of miR-429 and Bmi-1 in lung carcinoma (LC) tissues. METHODS: Samples of tumor tissues and paraplastic tissues were collected from 60 LC patients,January to December,2013. Expression of miR-429 and Bmi-1 mRNA in both tissues were detected by the quantitative real-time PCR (qPCR) assay. In addition,their relationships with clinical features and prognosis of the patients were analysed. RESULTS: The tumor tissues showed significantly decreased expression of miR-429 and increased expression of Bmi-1,compared with the paraplastic tissues (P < 0.01). Among the patients,those with lower expression of miR-429 and higher expression of Bmi-1 in their tumor tissues tended to have larger tumors,later clinical stages,more prone to lymph node metastasis and shorter survival time. CONCLUSION: The results show that the LC patients with lower miR-429 expression and/or higher Bmi-1 expression in tumour tissues frequently had worse clinical outcomes and poorer prognoses.

OBJECTIVE: To use a bioinformatics database for determining expression levels and clinical significance of FOXO3 in bladder cancer (BLCA). METHODS: The GEPIA databases were used to analyze the expression of FOXO3 gene in bladder cancer and normal bladder tissues. In addition,the RNA-Seq data and clinical data were downloaded from the UALCAN database and were used to determine correlations between FOXO3 gene expression and prognosis. TIMER was used to investigate correlations between FOXO3 and cancer immune infiltrate cells. The String database was used to identify proteins which interact with FOXO3. RESUITS: Based on analyses of the GEPIA database,expression of FOXO3 gene in bladder cancer tissues was significantly lower than that in normal tissues(P < 0.01). In addition,the low expression was significantly associated with longer and disease-free survival in patients than the high expression(all P < 0.05). Analyses of the TIMER database show that the FOXO3 expression had a positive correlation with infiltrating levels of B cells (r=0.12,P < 0.05),CD8⁺ T cells (r=0.262,P < 0.01),CD4⁺ T cells (r=0.12,P < 0.05),macrophages (r=0.252,P < 0.01),neutrophils (r=0.242,P < 0.01) and dendritic cells (r=0.162,P < 0.01) in bladder cancers. Analyses of protein-protein interactions reveal that AKT1,SIRT1,EP300,BCL2L11,SOD2,SGK1,AKT2,CREBBP,CDKN1B and SMAD4 interacted with FOXO3. CONCLUSION: Our findings demonstrate that the FOXO3 gene was expressed at low levels in bladder cancer tissues. However,high FOXO3 expression was correlated with poor prognosis of bladder cancer and was correlated with infiltrating levels of immune cells in bladder cancer.

OBJECTIVE: To explore relationships between differentially expressed miRNA-related single nucleotide polymorphism (SNPs) and the prognosis of gastric cancer,and to provide recommendations for molecular biomarkers for prognosis of gastric cancer. METHODS: Eleven differentially expressed miRNA-SNPs of gastric cancer were selected as gene loci by SNP microarray and bioinformatics methods. Single nucleotide polymorphisms (SNPs) of total DNA in peripheral blood leukocytes of 344 patients with gastric cancer were detected by Sequenom MassARRAY. An on the spot face to face interview was carried out to collect information on patient's condition,treatment,living and eating habits for one year after the illness. The information and survival analyses were used to study relationships between miRNA-related SNPs and prognosis of gastric cancer. RESULTS: Univariate analyses showed that the polymorphisms of rs9536676 of miR-1297 and rs17502941 of MSH2 were associated with the prognosis of gastric cancer,while the other polymorphisms had no significant association. Multivariate analyses showed that surgery,TNM stage and rs17502941 AA/AG genotype were independent risk factors for the prognosis of gastric cancer. According to stratification by age and TNM stages,rs9536676 of miR-1297,rs10413288 of miRNA-519b,rs7143252 of miR-379,rs1468063 of FAS and rs10277413 of EGFR were significantly associated with the survival distribution of patients with gastric cancer. The results showed that patients with rs9536676 AA/AG and rs17502941 GG,rs9536676 AA/AG and rs10413288 AA,rs17502941 GG and rs10413288 AA genotypes had increased risk of poor prognosis. CONCLUSION: The differential expression of certain miRNA-SNPs in gastric cancer was closely related to the prognosis of patients with gastric cancer. The association can be used to investigate mechanisms for predicting the prognosis.

OBJECTIVE: To study the expression of cAMP-response element binding protein (CREB), in esophageal squamous cell carcinoma (ESCC) tissues,and its relationship with clinical characteristics and prognosis of the patients. METHODS: Samples of tumor tissues and paraplastic tissues were collected from 60 cases of the ESCC patients from January to December,2010. Expression of CREB in both tissues and their relationships with the clinical feathers of the patients were analysed by using Western blotting and Immunohistochemical assays. RESULTS: The tumor tissues showed significantly increased expression of CREB protein,compared with the paraplastic tissues (P < 0.01). Among patients,those with higher CREB expression tended to have large tumors,later clinical stages,higher lymph node metastasis and shorter five-year overall survival rate with higher cumulative recurrence rate (P < 0.05). CONCLUSION: The ESCC tissues showed significantly elevated expression of CREB protein. Furthermore,the patients with higher CREB expression in tumor tissues frequently had worse clinical outcomes and poorer prognoses. Therefore,CREB has the potential to be an ESCC-associated tumor marker.

OBJECTIVE: To investigate impact of chromosome 1 translocation and locations of its breakpoints on semen quality in male carriers. METHODS: Lymphocytes in peripheral blood of patients with male infertility were cultured and G-banding technique was used for karyotype analysis. Semen examinations were performed according to WHO laboratory manual for the examination and processing of human semen. RESULTS: Twenty-nine male patients with infertility were diagnosed as carriers with chromosome 1 balanced translocations. Abnormal semen parameters were observed in most of patients (25/29,86%). The different degrees of abnormal semen quality were detected in the patients with the different translocation breakpoints. CONCLUSION: The chromosome 1 balanced translocations were associated with reduced semen quality. The degree of impact was closely related to the locations of the translocation breakpoints. Some molecular investigations are needed to better understand the impact of chromosome translocations on reproductive fitness.

OBJECTIVE: To explore the value of combined-biomarker detection method in early auxiliary diagnosis of silicosis. METHODS: Sixty-five silicosis male patients were chosen as the patient group,and seventy male workers without SiO2 dust exposure were selected as the control group by purposive method. Peripheral blood was collected from all subjects during the early morning before they had breakfast. Serum was separated from the blood samples. Levels of superoxide dismutase (SOD) and malondialdehyde (MDA) were determined by the colorimetric method,and levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were measured by enzyme-linked immunosorbent assay (ELISA). Taking the biomarkers (MDA,IL-6 and TNF-α) with significant changes in the serum of patients with stage I silicosis as the data set,ROC curves were drawn and the logistic regression model was used to evaluate the value of the combined biomarkers for early auxiliary diagnosis of silicosis. RESULTS: Compared with the control group,the SOD contents had no significant changes in the serum of patients with silicosis,while the MDA and IL-6 contents were significantly increased,and the TNF-α contents were significantly increased in the stage I silicosis patients. The results showed that the area under ROC curve was combined application (0.89) > IL-6(0.86) > MDA(0.81) > TNF-α(0.65). CONCLUSION: The combined application of IL-6,TNF-α and MDA improved the sensitivity and specificity of early auxiliary diagnosis of silicosis.