Inhibition of high-mobility group box 1 expression by siRNA in rat hepatic stellate cells

AIM:To explore the role of high-mobility group box 1 (HMGB1) protein during liver fibrogenesis and investigate the functional effects of HMGB1 gene silencing in hepatic stellate cells (HSCs) using siRNA.METHODS:Hepatic fibrosis in rats was induced through serial subcutaneous injections of dimethylnitrosamine,and expression of HMGB1 was detected by immunohistochemistry.HMGB1 siRNAs were developed and transiently transfected into HSC-T6 cells using Lipofectamine 2000.HMGB1 expression was evaluated by real-time polymerase chain reaction (PCR) and Western blotting analysis.Expression of α-smooth muscle actin (α-SMA) and collagen typesⅠand Ⅲ was evaluated by real-time PCR.Cell proliferation and the cell cycle were determined using the methyl thiazolyl tetrazolium method.Finally,collagen content in HSC supernatant was evaluated by an enzyme-linked immunosorbent assay.RESULTS:The results showed that HMGB1 was upregulated during liver fibrosis and that its expression was closely correlated with the deposition of collagen.siRNA molecules were successfully transfected into HSCs and induced inhibition of HMGB1 expression in a time-dependent manner.Moreover,HMGB1 siRNA treatment inhibited synthesis of α-SMA and collagen types Ⅰ and Ⅲ in transfected HSCs.CONCLUSION:This study suggests a significant functional role for HMGB1 in the development of liver fibrosis.It also demonstrates that downregulation of HMGB1 expression might be a potential strategy to treat liver fibrosis.

Detection of metastatic cancer cells in mesentery of colorectal cancer patients

AIM To detect the existence of isolated cancer cells in the mesentery of colorectum(named as Metastasis V), and investigate its clinical significance in colorectal cancer(CRC) patients.METHODS Sixty-three CRC patients who received radical excision between January 2012 and September 2015 were included. All the patients underwent laparoscopyassisted radical colorectomy or proctectomy [with complete mesocolic excision(CME) or total mesorectal excision(TME)] with R0 dissections at the Department of Gastrointestinal Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology. The location and size of the primary lesions were recorded immediately after the tumor was removed, with the surrounding mesenterium completely separated along the intestinal wall. Each dissected mesentery sample was analyzed for hematoxylin-eosin staining and immunohistochemistry using cytokeratin 19 antibody. Image Pro Plus Software 6.0(Media Cybernetics, CA, United States) was usedto semi-quantitatively measure the concentration of the cytokeratin 19 immunohistochemistry. The correlation between metastasis found in mesentery and clinicopathological characteristics was examined. The prognosis of patients was also evaluated by preoperative serum CEA level.RESULTS Metastasis V was detected in 14 of 63(22.2%) CRC patients who underwent laparoscopy-assisted radical colorectomy or proctectomy(with CME or TME) with R0 dissection in our hospital between January 2012 and September 2015. There was no significant difference in age, gender, tumor size, and tumor location in patients with Metastasis V(P > 0.05). Metastasis V was more likely to occur in poorly differentiated tumor(5/11; 45.5%) than moderately(8/46; 17.4%) and welldifferentiated one(1/6; 16.7%). The Metastasis V in N2 stage(9/14; 64.3%) was more frequent that in the N0 stage(3/35; 8.6%) or N1 stages(2/14; 14.3%). In addition, Metastasis V was positively related to the tumor invasive depth(T1:0/1, 0%; T2:1/12, 8.3%; T3:7/39, 17.9%; T4:6/11, 54.5%). Furthermore, preoperative serum CEA level in Metastasis V-positive patients was significantly higher than in Metastasis V-negative patients(4.27 ng/m L vs 3.00 ng/m L).CONCLUSION Metastasis V might be associated with a poor prognosis of CRC patients.

Unusual Patterns of Neural Tube Defects in a High Risk Region of Northern China

Objective To study the prevalence of different types of neural tube defects （NTDs） in Luliang Prefecture, Shanxi province, where the prevalence of NTDs is unusually high and the correlation between NTDs prevalence and patterns. Methods A surveillance population-based birth defects was performed in Luliang Prefecture, Shanxi province. Results The results of our study showed that the prevalence of NTDs was 2-fold higher in Luliang Prefecture than in other areas of Shanxi province. Unusual patterns of NTDs were found, however, multiple NTDs were relatively common in Luliang Prefecture, accounting for over 13% of all NTDs cases in China. Conclusion The prevalence of NTDs is associated with its patterns.

Association between Long Interspersed Nuclear Element-1 Methylation and Relative Telomere Length in Wilms Tumor

Background： DNA hypomethylation of long interspersed nuclear elements- 1 （LINEs- 1 ） occurs during carcinogenesis, whereas intbmaation addressing LINE-1 methylation in Wilms tumor （WT） is limited. The main purpose of our study was to quantity, LINE-1 methylation levels and evaluate their relationship with relative telomere length （TL） in WT. Methods： We investigated LINE-1 methylation and relative TL using bisulfite-polymerase chain reaction （PCR） pyrosequencing and quantitative PCR, respectively, in 20 WT tissues, 10 normal kidney tissues and a WT cell line. Significant changes were analyzed by t-tests. Results： LINE-1 methylation levels were significantly lower （P 〈 0.05） and relative TLs were sigmificantly shorter （P 〈 0.05） in WT compared with normal kidney. There was a significant positive relationship between LINE- 1 methylation and relative TL in WT （r = 0.671, P = 0.001 ）. LINE- 1 Methylation levels were significantly associated with global DNA methylation （r = 0.332, P 〈 0.01 ）. In addition, relative TL was shortened and LINE- 1 methylation was decreased in a WT cell line treated with the hypomethylating agent 5-aza-2＇-deoxycytidine compared with untreated WT cell line. Conclusion： These results suggest that LINE-1 hypomethylation is common and may be linked to telomere shortening in WT.