Objective To explore the expression of the Pendrin gene (SLC26A4) and protein in multinodular goiter.Methods Thyroid tissues were obtained from 40 multinodular goiter patients undergoing surgery while the control group were obtained from 40 nomal thyroid tissues.RT-PCR was used to test SLC26A4 gene while western blot and immunohistochemistry were used to test Pendrin protein expression and distribution.Results SLC26A4 mRNA expression in multinodular goiter tissue was significantly increased in comparison with normal nodular tissues (t=2.663,P=0.011).Pendrin protein expression in multinodular goiter group was higher than that in normal tissue (t=2.286,P=0.026).The immunohistochemistry results showed that the Pendrin protein in multinodular goiter was mainly located in cytoplasm.There was positive expression in 24 patients (60％) in multinodular goiter group,while it was in 14 patients (35％) in the normal control group.The difference was significant (X2=5.013,P=0.025).Pendrin protein mainly expressed in cytoplasm in multinodular goiter tissue while it was mainly in cytomembrane in the normal control group.Conclusion SLC26A4 mRNA and its coding protein Pendrin expression are increased in multinodular goiter group,and mainly located in cytoplasm,indicating that iodide transporter function may be damaged when multinodular goiter occurs.

Objective To explore the correlations between expression of miRNA-21 and PTEN and the invasion, metastasis of colorectal carcinoma.Methods The mRNA level of miRNA-21 and mRNA and protein levels of PTEN were assayed by real-time quantitative PCR and immunohistochemical methods respectively in 65 colorectal carcinoma specimen.Results The expression of miRNA-21 mRNA in cancer tissues was significantly higher than that in paracancerous tissues (t =3.50, P ＜ 0.05).Both mRNA and protein expressions of PTEN in tumor tissue were significantly lower than that in paracancerous tissues (t =7.35,t =12.23;P ＜ 0.05).Expression of miRNA-21 mRNA, PTEN mRNA and protein were obviously related with depth of invasion(F =18.36 ,F =17.26 ,F =12.83;P ＜ 0.05), Dukes stage (F =31.25, F =24.43, F =57.12;P ＜ 0.05) and lymph node metastasis (t =5.85, t =2.18, t =4.05;P ＜ 0.05), while it was not related to tumor differenciation and tumor location (F =7.39, t =4.62;F =7.78, t =1.29;F =5.14,t =1.37;P ＞ 0.05).Positive relation between miRNA-21 and PTEN in colorectal carcinoma was identified (r =-0.994, r =-0.927;P ＜ 0.05).Conclusion High expression of miRNA-21 and low expression of PTEN are both closely associated with invasion and metastasis of colorectal carcinoma.

Aim To investigate the time-dependent effect of insulin on the expression of SREBP-1(sterol regulatory element binding protein-1),FAS(fat acid synthase)and lipid droplet formation in HKC cells(human renal proximal tubular epithelial cells line).MethodsHKC cells were respectively treated with 100 nmol·L~(-1) insulin for 0,2,4,6,12 h and 24 h.The analysis of SREBP-1 and FAS mRNA was performed by RT-PCR and the expression of SREBP-1 protein was detected by Western blot and immunocytochemistry.Furthermore,Oil Red O staining was used to determine cellular lipid droplet formation.ResultsCompared with HKC cells of 0 h group,there was no difference of SREBP-1 and FAS mRNA in HKC cells of 2 h group.However,the expression of SREBP-1 and FAS mRNA was significantly increased in HKC cells of 4,6 h and 12 h group.Further,the most expression of SREBP-1 and FAS mRNA was at 6 h group and was respectively increased by 3.578 and 4.272 times compared with 0 h group.The results of Western blot showed that the precursor and mature segments of SREBP-1 protein in 4,6 h and 12 h group HKC cells were increased and those of 6 h group HKC cells were the highest and about 4.106 and 5.167 times than those of 0 h group HKC cells.Immunocytochemistry presented the result that SREBP-1 protein was located in the plasma and the expression of 4,6 h and 12 h group HKC cells was significantly higher than that of 0,2 h and 24 h group HKC cells.The result of Oil Red O staining showed that lipid droplet markedly deposited in 6 h group HKC cells,contrarily,no lipid droplet was found in HKC cells of other groups.ConclusionAbove results suggested that insulin up-regulated SREBP-1 and FAS in time-dependent manner that led to cellular lipid droplet deposit,which may play an important role in the pathogenesis of renal lipid accumulation in metabolism syndrome.

Purpose To investigate the role of PTEN in podocyte injury in patients with diabetic nephropathy ( DN) . Methods Uri-nary samples from 30 patients with DN and 10 healthy volunteers were collected to detect the level of PCX by ELISA. Renal biopsies were reviewed to observe the morphological changes. All patients with DN were divided into three groups by glomerular lesion. The ex-pression of p-Akt and PTEN in glomeruli was detected by immunohistochemistry. Results The levels of PCX in the urine were signifi-cantly higher in patients with DN compared with those in healthy volunteers, and gradually increased along with glomerular lesion aggra-vating. The expression of p-Akt and PTEN increased in patients with DN compared with healthy volunteers. Although the expression of p-Akt and PTEN decreased with the aggravation of glomerular lesion, they were still higher than that in volunteers. There were obvious-ly positive correlation between the level of PCX and 24-h urinary protein and negative correlation between the level of PCX and the ex-pression level of p-Akt and PTEN. Conclusion PTEN down-regulation may be associated with podocyte injury in DN, which may be associated with the phosphorylation of Akt.

Objective:To investigate the expression of Agmatinase (AGMAT) in breast cancer and its effect on the proliferation, migration and invasion of breast cancer cell lines.Methods:The expression levels of AGMAT in cancer and adjacent tissues of 1 094 breast cancer samples in The Cancer Genome Atlas (TCGA) database were analyzed. And its expression degree was verified in breast cancer cell lines MDA-MB-231, MCF-7, HCC-1937 and T-47D. The expression of AGMAT in breast cancer cells was knocked down by shRNA, and the expression level of AGMAT mRNA was detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), and the protein expression level was detected by Western blotting assay. Cell count and MTT assay were used to detect cell proliferation. Cell apoptosis and cell cycle changes were detected by flow cytometry. Cell migration ability was detected by cell scratch assay, cell invasion ability was detected by Transwell assay, and the expression level of related proteins in cells was detected by Western blotting assay. Measurement data were expressed as mean ± standard deviation ( ± s), comparisons between multiple groups were performed using ANOVA, pairwise comparisons were performed using independent samples t-test, and Tukey′s post hoc multiple test was used. Results:Analyzing the data of breast cancer samples in TCGA database, it was found that the expression level of AGMAT in cancer tissues was significantly higher than that in adjacent tissues ( FC=10.537, P<0.001). Flow cytometry showed that knocking down the expression of AGMAT inhibited the proliferation of breast cancer cells, and induced cell apoptosis [(3.20±0.10)% vs (6.83±0.06)%, t=62.35, P<0.001], and caused G 1 cell cycle arrest [(49.51±2.22)% vs (31.44±1.67)%, t=42.56, P=0.001]. The results of cell scratch assay and Transwell assay showed that decreased GMAT expression could reduce the cell migration ability [(34.27±1.67) % vs (57.97±0.58) %, t=33.52, P<0.001], invasive ability (163.00±1.77 vs 61.00±0.74, t=52.50, P<0.001). The results of Western blotting assay showed that the protein expression levels of Twist, Vimentin, matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) decreased, while the protein expression level of E-cadherin increased. AGMAT was involved in the process of epithelial-mesenchymal transition in breast cancer. Conclusion:AGMAT can be used as a prospective biomarker affecting the invasion, metastasis and therapeutic targets of breast cancer.

Objective: To analyze the genetic characteristics of Coxsackievirus A4 isolated from Taian, 2017-2018. Methods: Sixty throat swab samples of the children who visited Taian Maternal and Child Health Hospital during the year 2017-2018 and were diagnosed as hand, foot and mouth disease, were collected and aseptically inoculated. Fluorescent quantitative PCR analysis was performed using the universal primer for enteroviruses. The high-throughput sequencing was applied to the enterovirus-positive samples, and the full-length genome sequences of the viruses were obtained. Phylogenetic analysis was performed using Mega5.05 and RaxML respectively, and sequence homology and amino acid mutation sites were also analyzed using Mega5.05. Results: Four whole genome sequences of CV-A4 isolated from infants aged 17-19 months old were obtained. Phylogenetic analysis of the full length CV-A4 genomes showed that apart from MG550920/AA/Henan/2016, the remaining CV-A4 strains from China (97.2%), including the four strains from Taian, fell within Group 3. The VP1 genes could be classified into four genotypes and 98.5% of the Chinese strains belonged to genotype D, and the four strains from Taian belonged to D2. It was notable that the Taian isolate A1/Taian is closely related to two strains C179 and C062 from Australia both in the complete genome and the VP1 gene, as well as one strain YT184R isolated from Yantai in 2016 by us. Compared with the prototype CV-A4 strain High Point, 18 amino acid mutations were found in the P1 region. Conclusions Both phylogenetic trees estimated using the complete genome and the VP1 gene sequences revealed that the four CV-A4 isolates from Taian fell within the same clade with the majority of CV-A4 strains circulating in China. Compared with the prototype CV-A4 strain, several amino acid variations have occurred in the P1 region, which warrants further investigation.