Differentiated embryonic chondrocyte gene 2 (DEC2) inhibits transdifferentiation of mouse glomerular endothelial cells and renal fibrosis by blocking TGF-β/ROCK1 signaling pathway / 细胞与分子免疫学杂志

Objective To explore the protective mechanism of transdifferentiation of glomerular endothelial cells based on the differentiated embryonic chondrocyte gene 2 (DEC2) via the TGF-β/ROCK1 signaling pathway. Methods The 24 mice were randomly divided into sham group, UUO group, UUO combined with vector group and UUO combined with DEC2 group, with 6 mice in each group. A unilateral ureteral obstruction (UUO) model was established in each group, except for the sham group. In the UUO combined with vector group and UUO combined with DEC2 group, 10 μL (108 PFU) of vector or DEC2 was injected into each kidney on day 0 (immediately after UUO) under the guidance of the ultrasound system. The mice were sacrificed 14 days after the operation, and the kidneys were collected for histological examination and Western blot analysis: HE staining was used to observe the histological changes of kidneys, Masson staining to observe the renal fibrosis, and Western blot analysis to detect the protein expression. In vitro, normal human glomerular endothelial cells (GEnCs) was selected as the research objects. GEnCs stimulated with TGF-β were treated with ROCK1 inhibitor Y-27632 or DEC2 transfection. Western blot analysis was used to detect the expression of ROCK1, α-SMA, DEC2 and E-cadherin in GEnC exposed to transforming growth factor β (TGF-β). The localization of ROCK1 and DEC2 in GEnCs cells was detected by immunofluorescence cytochemistry. The relationship between the ROCK1 and DEC2 was confirmed by co-immunoprecipitation. Results Compared with the sham group, the UUO groups showed significant renal fibrosis and collagen accumulation on the 14th day. In the UUO groups, the expression of DEC2 and E-cadherin in the kidney tissue of the mice was significantly reduced, and the expression of α-SMA significantly increased. Compared with the UUO combined with vector group, the kidney fibrosis and collagen accumulation in the UUO combined with DEC2 group decreased, and the expression of ROCK1 and α-SMA decreased and the expression of DEC2 and E-cadherin increased in the kidney tissue. TGF-β enhanced the expression of ROCK1 and α-SMA in GEnCs cells in a time-dependent manner, and the levels of DEC2 and E-cadherin decreased. Treatment with the ROCK1 inhibitor Y-27632 partially abrogated the TGF-β-induced increase in the expression of ROCK1 and α-SMA and decrease in the expression of DEC2 and E-cadherin. In addition, transfection of GEnCs cells with DEC2 before TGF-β stimulation reduced the expression of ROCK1 and α-SMA, and increased the expression of DEC2 and E-cadherin. Immunofluorescence cytochemical staining showed that DEC2 co-localized with ROCK1 in GEnCs, and the co-immunoprecipitation showed that DEC2 and ROCK1 pulled down each other. Conclusions DEC2 is down-regulated in fibrotic renal tissue, while up-regulated DEC2 inhibits epithelial myofibroblast transdifferentiation and renal fibrosis of GEnC by blocking TGF-β/ROCK1 signaling pathway.

Single lung transplantation assisted by extracorporeal membrane oxygenation technique duringperioperative period:a report of 6 cases / 中华器官移植杂志

Objective To summarize the application experiences and curative efficacies of single lung transplantation assisted by extracorporeal circulation with coated lung ,centrifugal pump and coated pipe .Methods Retrospective analysis was conducted for clinical data of 6 adult patients with respiratory insufficiency undergoing single lung transplantation .The changes of hemodynamics and oxygenation before and after adjuvant treatment were observed ,the effects of adjuvant evaluated and the experiences of application summarized .Results The hemodynamic parameters post-assistance significantly improved as compared with that pre-assistance and pulmonary arterial pressure dropped from (56 ± 15 ) to (45 ± 13 ) mmHg with statistically significant differences . Arterial blood gas parameters significantly improved .PO2 spiked from (47 ± 12) to (68 ± 9) mmHg and PCO2 declined from (65 ± 14) to (55 ± 12)mmHg .And there were statistically significant differences .All patients were discharged successfully .Conclusions The simple extracorporeal membrane oxygenation system of coated lung , centrifugal pump and coated pipe during routine extracorporeal circulation may guarantee the operative safety of single lung transplantation and provide a new therapeutic option .

Promoter methylation and protein expression of tumor suppressor candidate 1 gene in adolescent papillary thy-roid carcinoma / 中华实用儿科临床杂志

Objective To study the relationship between status of methylation of tumor suppressor candidate 1 gene(TUSC1) promoter and expression of its protein in adolescent papillary thyroid carcinoma(PTC). Methods Forty cases of adolescent PTC were chosen and the corresponding para carcinoma tissues were taken from July 2010 to Decem-ber 2013 in the First Affiliated Hospital of Zhengzhou University surgical specimens of the thyroid gland and were con-firmed by pathology. Male 12 cases,female 28 cases,median age 14 (10-18) years old. Tumor node metastasis (TNM) stageⅠ-Ⅱ13 cases,Ⅲ-Ⅳstage 27 cases;gradeⅠin 15 cases,gradeⅡin 25 cases;lymph node metastasis in 22 cases,18 cases were negative. Methylation-specific polymerase chain reaction (MSP) and Western blot were applied respectively to examine the methyaltion of TUSC1 gene promoter and its protein expression of 40 samples of adolescent PTC and their matched adjacent non-cancerous epithelium. Results The results of MSP revealed that there was no methylation of TUSC1 gene promoter in adjacent non-cancerous epithelium,while in the adolescent PTC,the hyper-methylation rate was 60%(24/60 cases,χ2=34. 28,P<0. 05). In additional,it was related to the TNM stage,pathological grade and lymph node metastasis (χ2=4. 862,7. 111,5. 625,all P<0. 05). The result of Western blot revealed that the positive expression rate of TUSC1 protein was 100% in adjacent non-cancerous epithelium and 30%(12/40 cases) in adolescent PTC (χ2=14. 118,P<0. 05),which was related to the TNM stage,pathological grade and lymph node metastasis (χ2=5. 215,6. 222, 5. 079,all P<0. 05). There was distinct correlation between methylation of TUSC1 gene promoter and the protein expres-sion (r=-0. 84,P<0. 05). Conclusions Methylation of promoter might be one of the important mechanisms of inactiva-tion of TUSC1 gene,and might play an important role in carcinogenesis and progression of adolescent PTC.