Tension promotes keloid fibrosis: a preliminary study / 中华皮肤科杂志

Objective:To preliminarily evaluate the effect of tension stimulation on the biological activity of and expression of fibrosis marker genes in keloid fibroblasts (KD-Fbs) .Methods:Three patients who were diagnosed with keloids and received surgical treatment were collected from the Department of Dermatology, Tangdu Hospital, the Fourth Military Medical University from January to March 2017. Human KD-Fbs were isolated from resected keloid tissues, and subjected to primary culture. The third- to sixth-passage KD-Fbs were divided into tension group and control group to be cultured in the tension-based chamber and control chamber respectively, and subjected to tension stimulation and normal culture respectively. Cell counting kit-8 (CCK8) assay was performed to assess the proliferative activity of KD-Fbs after 1-, 2-, 3- and 4-day culture, and the scratch assay to evaluate the migratory ability of KD-Fbs after 1- and 2-day culture. After 48-hour treatment, real-time quantitative PCR and Western blot analysis were performed to determine the mRNA and protein expression of fibrosis markers type Ⅰ collagen, fibronectin and α-smooth muscle actin (α-SMA) in KD-Fbs respectively. Two-independent-sample t test was used for comparisons between 2 groups. Results:CCK8 assay showed that the proliferative activity of KD-Fbs was significantly higher in the tension group than in the control group after 1-, 2-, 3- and 4-day culture ( t=3.05, 7.00, 16.65, 15.19, respectively, all P< 0.05) . After 1- and 2-day culture, the scratch assay showed that the migration rate of KD-Fbs was significantly higher in the tension group (48.65%±3.96%, 100.00%, respectively) than in the control group (9.36%±1.14%, 50.35%±4.23%, t=16.53, 20.35, respectively, both P< 0.01) . Real-time quantitative PCR showed that the mRNA expression of type Ⅰ collagen, fibronectin and α-SMA was significantly higher in the tension group (3.04±0.20, 2.16±0.10, 3.76±0.24, respectively) than in the control group (1.00; t=17.57, 21.01, 20.25, respectively, all P< 0.01) . As Western blot analysis revealed, changes in the protein expression of the 3 fibrosis markers were consistent with their mRNA expression changes (all P< 0.05) . Conclusion:Tension may participate in the fibrosis in keloids by promoting the expression of fibrosis marker genes, and enhancing the proliferative and migratory ability of KD-Fbs.

High expression of DNMT3B promotes proliferation and invasion of hepatocellular carcinoma cells Hippo signaling pathway / 南方医科大学学报

OBJECTIVE@#To explore the role of DNMT3B in regulating the proliferation and invasion of hepatocellular carcinoma (HCC) cells.@*METHODS@#We collected the tumor tissues and adjacent tissues from a total of 175 patients with HCC diagnosed in the Second Affiliated Hospital of Chongqing Medical University between May, 2008 and May, 2013 to prepare the tissue microarrays. The association of the expression of DNMT3B with the prognosis and the tumor-free survival and tumor-specific survival rates of the patients was analyzed. Univariate and multivariate Cox regression analyses were used to analyze the effect of DNMT3B expression on the prognosis of HCC. We used RNA interference technique to knock down the expression of DNMT3B in Huh-7 hepatoma cells and observed the changes in cell proliferation using CCK-8 assay and EDU staining and in cell migration and invasion ability using Transwell assay.@*RESULTS@#The positive rates of DNMT3B was significantly higher in HCC tissues than in paired adjacent tissues (67.4% 41.1%, =0.015). A high DNMT3B expression in HCC was significantly associated with the tumor size (=0.001), vascular invasion (=0.004), and intrahepatic metastasis (=0.018). The patients with high DNMT3B expressions had significantly lower tumor-free and tumor-specific survival rates than those with low DNMT3B expressions ( < 0.005). In Huh-7 cells, silencing DNMT3B significantly inhibited the cell proliferation and inhibited cell migration and invasion. Western blotting showed that silencing DNMT3B obviously increased LATS1 expression, decreased the expression of YAP1, and activated Hippo signaling pathway. Methylation-specific PCR showed that the methylation level of LATS1 was decreased in the cells with DNMT3B silencing.@*CONCLUSIONS@#The expression level of DNMT3B is significantly higher HCC tissues than in the adjacent tissues, and the high expression of DNMT3B is closely related to the low survival rate of the patients. Silencing DNMT3B inhibits the proliferation, migration and invasion of HCC cells. DNMT3B promotes the progression of HCC primarily by enhancing the expression of YAP1 through methylation of LATS1 and inhibition of its expression, which inhibits the anti-cancer effect of Hippo signaling pathway.

Clinical analysis of ano-saving surgery in low rectal carcinoma / 中国普通外科杂志

ObjectiveTo evaluate the indication, the operation pattern and therapeutic effect of the ano-saving surgery in low rectal carcinoma. MethodsAccording to anorectal finger-examination, IRUS,and CT, 94 out the 161 rectal cancer patients were selected to have ano-saving surgery from August 1993 to December 1994.Excreting function, 5-year survival rate and local recurrence rate of the various operation were compared. ResultsThe perfomed rate of ano-saving operation in low rectal cancer was 58.4%. Among them, low anastomosis was done in 6 cases, ultra-low anastomosis in 48 cases,Park′s operation in 25 cases,and Bacon operation in15 . The death rate was 2.1%(2 cases).Incidence of anastomotic leakage after the surgery was 3.2%(3 cases), and only 13 cases had anastomotic narrowing(13.8%) within 1 year. The successful rates of excreting function after the surgery were respectively as follows: low anastomosis 100%, ultra-low anastomosis 97.9%,Park′s surgery 88.0%,and Bacon surgery 53.3%. The 5 year survival rates and the local recurrence rates were respectively,as follows: low anastomosis 83.3% and 0; ultra-low anastomosis 79.2% and 4.2%; Park′s 64.0% and 12.0%; and Bacon 66.7% and 13.3%,respectively. ConclusionsUltra-low colo rectum anastomosis becomes the main operative pattern to preserve anal sphincter in low rectal cancer.There is no difference in the 5-year survival rate and the excreting function among low, ultra-low anastomosis and Park′s operation, but the low and ultra-low colo-rectum anastomosis were obviously better than that of Bacon and Miles operation.The local recurrence rates of low and ultra-low colon-rectum anastomosis are lower than that of Miles′.There is no difference in the 5-year survival rate and local recurrence rate between Park′s, Bacon and Miles operation.