Objective To explore the dignostic value of the candidate molecular markers in Urothelium carcinoma based on Illumina Transcriptome Sequencing. Methods Candidate genes which were screened with transcriptome sequencing were validated by using real-time fluorescent quantitative PCR in 31 pairs of tumor and normal tissues of Urothelium carcinoma patients from Affliated Hai kou Hospital Xiangya School of Medicine Central South University. Results The four candidate genes (CDH1,VEGFA,PTPRF and CLDN7) in tumor tissues were up-regulated compared with normal groups in samples of sequencing. The relative expression of VEGFA mRNA were higher than that in normal tissues. There were significant difference in two groups (P<0.05). PTPRF and CLDN7 were also up-regulated, but no significant difference was found (P > 0.05). PTPRF was closely related with the recurrence of tumor (P=0.002), and the predict sensitivity and speciality rate were 90.0%and 83.3%respectively. Conclusions CDH1 and VEGFA play important roles in the occurrence and development of bladder urothelial carcinoma, and they may be the possible biomarkers. PTPRF is expected to be a molecular reference recurrent predictor of bladder tumor.

Objective To explore the pathogenic mechanism by detecting the expression of membrane glycoprotein in the platelets of nonmuscle myosin heavy chain 9 related disease (MYH9-RD)patients.Methods Periperal bloods were obtained from 11 MYH9-RD patients and 7 normal family members.Flow cytometry was used for detecting the expression of the platelet membrane glycoprotein including GP Ⅱ b/Ⅲa(CD41/61),GP Ⅰ a(CD49b),GP Ⅰ b/Ⅸ/Ⅴ (CD42a) GP Ⅰ b(CD42b) and GPⅣCD36).Results The expression fluorescence intensity of platelet membrane glycoprotein GP Ⅱ b/Ⅲ a CD41/61),GPⅠa(CD49b),GP Ⅰ b/Ⅸ/Ⅴ (CD42a) GP Ⅰ b(CD42b) and GPⅣ (CD36) are 653.7 ±192.7,420.0 ± 151.3,667.7 ± 371.3 and 236.4 ± 64.2 respectively,which are significantly higher than those in normal controls (406.7 ± 126.1,181.2 ± 29.3,271.4 ± 91.6 and 136.1 ± 23.5 ; P ＜ 0.01) ; The expression of GP Ⅰ a(CD49b) was lower in patients with MYH9-RD (139.1 ± 54.9) than that in normal controls (192.2 ± 143.4),but there was no significant difference (P ＞ 0.05).Conclusion In our study,the diverse clinical manifestations in patients with MYH9-RD is probably associated with the expression level of platelet membrane glycoprotein

Objective To detect the expression of humar telomerase reverse transcriptase (hTERT) and telomerase activity in HeLa,MCF-7,SMMC7721 PC-3m and U2OS cell lines. Methods The techniques of immunochemistry and TRAP-ELISA were employed to detect the expression of hTERT and telomerase activity in different tumor cell lines.Results The positive rate of hTERT in HeLa cells(93.75%?3.10%)was significantly higher than that in U2OS cells(2.75%?0.96%),besides the other three cell lines showed an positive rates of hTERT in MCF-7(92.50%?2.65%),SMMC7721 (53.75%?2.22%)and PC-3m(23.50%?2.89%); Meanwhile,the telomerase activity of HeLa cells (94.58%?3.49%) was also much higher than that of U2OS(3.02%?0.43%),likewise the telomerase activities of the other three cell lines (MCF-7,SMMC7721,PC-3m) were 73.90%?4.50%,66.67%?3.35% and 50.62%?1.96%, respectively. Conclusion The expression of hTERT and telomerase activity show obvious differences among five tumor cell lines,suggesting that telomerase inhibitors cannot effect on all the tumor cell lines.

This paper describes a practical process for a SGLT2 inhibitor dapagliflozin. The target product was synthesized from 1-chloro-2-( 4-ethoxybenzyl)-4-iodobenzene and 2, 3, 4, 6-tetra-O-acetyl-alpha-D-glucopyranosyl bromide by iodine-magnesium exchange, and coupling and acetyl removing reactions with the total yield of 50%. This practical process highlights fewer reaction steps, less waste and mild reaction conditions.

Objective:To investigate the effect of interleukin-1β (IL-1β) on epithelial-mesenchymal transition (EMT) and cytoskeleton rearrangement of renal tubular epithelial cells.Methods:Immortalized renal tubular epithelial cell line NRK52E was cultured in vitro with IL-1β (30 μg/L) for 3 days and 6 days,then the cell morphology was observed;The mRNA expressions of α-smooth muscle actin (α-SMA),cytoskeleton components β-actin and α-tubulin were semi-quantitative examined by RT-PCR.The protein expression of α-SMA and arrangements of β-actin and α-tubulin were assessed by immunofluorescent staining.Results:After induced by IL-1β for 3 days and 6 days in vitro,the mRNA and protein expression of α-SMA increased significantly compared with corresponding control cells (P<0.001),it prompted that NRK52E cells underwent EMT;At the same time,the cell morphology also changed,from a typical multilateral paving stone to fibroblast-like appearance,with multiple processes; Cytoskeletal protein β-actin mRNA expression was also slightly increased (P<0.05).The distributions and arrangements of β-actin protein were also changed,from cell membrane transferred to peri-nucleus and cytoplasm,moreover it formed fiber bundle-like structures.However,another cytoskeleton protein α-tubulin in IL-1β induced cells,neither it's mRNA expression nor it's distribution had significant differences compared with the control group.Conclusion:IL-1β can induce NRK52E cells undergoing EMT in vitro,cell morphology changes into fibroblast-like appearance with multiple processes,and also the cytoskeleton protein β-actin expression increases and rearrangement occurrs.However,there was no changes onα-tubulin.

Objective To study the combined effect of interleukin-21 gene transfer and ionizing radiation on the growth of cervical carcinoma HeLa cells.Methods Previously constructed Ad-IL-21 gene was amplified by infecting 293A cells and the titer was measured by TCID50 method. HeLa cells were transfected with Ad-1L-21 and then irradiated with 6 Gy 137Cs γ-rays.The cells were divided into 5 groups,including blank control,Ad-LaeZ group,Ad-IL-21 group,radiation group and Ad-IL-21 combined with radiation group (combination group).The cell growth,cell cycle,apoptosis,and the expressions of IL-21 gene and protein in HeLa cells were detected.Results Ad-IL-21 was successfully amplified and the titer of Ad-11.-21 was 9 × 1010 pfu/ml.Compared with Ad-IL-21 group and radiation group,the cell growth of combination group was significantly inhibited at 96 h after transfection ( F =85.26,72.98,P ＜ 0.05 ).The cells in combination group were arrested in G1 phase and decreased at S phase( F =36.69,34.83,P ＜ 0.05),while the cellular apoptosis increased markedly ( F =28.23,25.57,P ＜ O.05 ). The gene expression of 1L-21 in the combination group was 1.54- and 2.43-fold of Ad-IL-21 group and blank control group,respectively (F=22.31,36.65, P ＜ 0.05 ), while the protein expression of IL-21 in the combination group was 1.62-fold and 2.31-fold of Ad-IL-21 group and blank control group,respectively ( F =27.36,35.86,P ＜ 0.05 ).Conclusions Ad-IL-21 gene transfection combined with radiation has synergic effect on the inhibition of cervical carcinoma cell growth.

OBJECTIVETo report on a rare D13S317-off-ladder (OL) allele identified in an ethnic Han Chinese from Hainan.

METHODSThe rare allele D13S317-OL was detected with a short tandem repeat (STR)-PCR method and DNA sequencing.

RESULTSThe core repeat sequence of the rare allele was found to be [TATC]5.

CONCLUSIONThe rare D13S317-5 allele also exists among ethnic Hans from Hainan, which may have a significant value for personal identification and paternity testing.

Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Female ; Humans ; Male ; Microsatellite Repeats ; genetics ; Paternity ; Polymerase Chain Reaction ; methods