Minimally invasive endoscopic repair of rectovaginal fistula.

BACKGROUND: Surgical techniques for repair of rectovaginal fistula (RVF) have been continually developed, but the ideal procedure remains unclear. Endoscopic repair is a novel and minimally invasive technique for RVF repair with increasing reporting. AIM: To review the current applications and preliminary outcomes of this technique for RVF repair, aiming to give surgeons an alternative in clinical practice. METHODS: Available articles were searched according to the search strategy. And the sample size, fistula etiology, fistula type, endoscopic repair approaches, operative time and hospital stay, follow-up period, complication and life quality assessment were selected for recording and further analysis. RESULTS: A total of 11 articles were eventually identified, involving 71 patients with RVFs who had undergone endoscopic repair. The principal causes of RVFs were surgery (n = 51, 71.8%), followed by obstetrics (n = 7, 9.8%), inflammatory bowel disease (n = 5, 7.0%), congenital (n = 3, 4.2%), trauma (n = 2, 2.8%), radiation (n = 1, 1.4%), and in two patients, the cause was unclear. Most fistulas were in a mid or low position. Several endoscopic repair methods were included, namely transanal endoscopic microsurgery, endoscopic clipping, and endoscopic stenting. Most patients underwent > 1-year follow-up, and the success rate was 40%-93%, and all cases reported successful closure. Few complications were mentioned, while postoperative quality of life assessment was only mentioned in one study. CONCLUSION: In conclusion, endoscopic repair of RVF is novel, minimally invasive and promising with acceptable preliminary effectiveness. Given its unique advantages, endoscopic repair can be an alternative technique for surgeons.

Autophagy as a Vital Therapy Target for Renal Cell Carcinoma.

Autophagy is a process that degrades and recycles superfluous organelles or damaged cellular contents. It has been found to have dual functions in renal cell carcinoma (RCC). Many autophagy-related proteins are regarded as prognostic markers of RCC. Researchers have attempted to explore synthetic and phytochemical drugs for RCC therapy that target autophagy. In this review, we highlight the importance of autophagy in RCC and potential treatments related to autophagy.

Emerging roles of hsa_circ_0005075 targeting miR-431 in the progress of HCC.

Circular RNAs (circRNAs) are non-coding RNAs that play key roles in the pathogenesis of diseases and are associated with human hepatocellular carcinoma (HCC). Previous reports have shown the circRNA hsa_circ_0005075 is highly expressed in HCC tissues, but its function is unknown. In this study, we confirmed circ_0005075 is significantly increased in HCC tissues and cell lines. This overexpression was associated with increased numbers of proliferative, migrated and invasive SMMC-7721 cells. In addition, hsa_circ_0005075 inhibited the transcription activity of miR-431 measured by dual-luciferase reporter assays. In contrast, silencing hsa_circ_0005075 decreased cell number. Finally, effects after hsa_circ_0005075 silencing were rescued by co-transfection with miR-431 inhibitor. These results suggest hsa_circ_0005075 promotes HCC via miR-431 regulation.

The biological roles and clinical implications of microRNAs in clear cell renal cell carcinoma.

Clear cell renal cell carcinoma (ccRCC) accounts for about 3% of tumors in adults as well as 85% of all primary renal carcinoma. And it is the third most predominant urological carcinoma, but it has the maximum mortality rate. Early diagnosis and proper follow-up of ccRCC patients may improve the prognosis of the illness. Thus, it is imperative to understand the new biomarkers of ccRCC and study new method for the modern therapy of this deadly disease. Furthermore, a large number of microRNAs (miRNAs), small non-coding RNAs, have been relevant to tumor type, stage, or survival and miRNAs might be progressed as the markers of diagnosis or prognosis in ccRCC. A growing body of data also advised the rationality of regarding miRNAs as therapeutic targets for ccRCC treatment. In this review, we tried to summarize biogenesis of miRNAs and their expression profiles, biological roles, and clinical implications in ccRCC.

Therapeutic potential of cysteine-rich protein 61 in rheumatoid arthritis.

Cysteine-rich protein 61 (Cyr61)/CCN1, a product of an immediate early gene, can directly accommodate cell adhesion and migratory processes whilst simultaneously regulating the production of other cytokines and chemokines through paracrine and autocrine feedback loops. This intricate functionality of Cyr61 indicate its important role in targeting components of the infectious or chronic inflammatory disease processes including rheumatoid arthritis (RA). Recent work has focused on the role of Cyr61 in RA. For example, Cyr61 induced proIL-1ß production in FLS via the AKT-dependent NF-κB signaling pathway. Moreover, Cyr61-siRNA decreased the levels of matrix metalloproteinase (MMP)-3 and MMP-13, and induced apoptosis in RA-FLS cells. These results indicated that Cyr61 may represent a novel target for the treatment of RA. In this article we will introduce the molecular properties of Cyr61, discuss the function of Cyr61, and the therapeutic potential of modulating the Cyr61 in RA.

NLRC5 promotes cell proliferation via regulating the AKT/VEGF-A signaling pathway in hepatocellular carcinoma.

NLRC5, a newly found member of the NLR family and the largest member of nucleotide-binding, has been reported to regulate immune responses and is associated with hepatocellular carcinoma (HCC). We investigated the mechanisms and signaling pathways of NLRC5 in HCC progression. Increased expression of NLRC5, vascular endothelial growth factor-A (VEGF-A) were found in human HCC tissue. There was a positive correlation between NLRC5 and VEGF-A expression and cell proliferation were enhanced in NLRC5-overexpressing HepG2 cells, but inhibited in cells with NLRC5 silencing treatment. Interestingly, we found that up-regulation of NLRC5 also coordinated the activation of PI3K/AKT signaling pathway. An AKT inhibitor LY294002 blocked VEGF-A expression and AKT phosphorylation in HepG2 cells and NLRC5-overexpressing HepG2 cells. These results demonstrate that NLRC5 promotes HCC progression via the AKT/VEGF-A signaling pathway.

Cryptolepine derivative-6h inhibits liver fibrosis in TGF-ß1-induced HSC-T6 cells by targeting the Shh pathway.

Liver fibrosis is a worldwide problem with a significant morbidity and mortality. Cryptolepis sanguinolenta (family Periplocaceae) is widely used in West African countries for the treatment of malaria, as well as for some other diseases. However, the role of C. sanguinolenta in hepatic fibrosis is still unknown. It has been reported that Methyl-CpG binding protein 2 (MeCP2) had a high expression in liver fibrosis and played a central role in its pathobiology. Interestingly, we found that a cryptolepine derivative (HZ-6h) could inhibit liver fibrosis by reducing MeCP2 expression, as evidenced by the dramatic downregulation of α-smooth muscle actin (α-SMA) and type I collagen alpha-1 (Col1α1) in protein levels in vitro. Meanwhile, we also found that HZ-6h could reduce the cell viability and promote apoptosis of hepatic stellate cells (HSCs) treated with transforming growth factor beta 1(TGF-ß1). Then, we investigated the potential molecular mechanisms and found that HZ-6h blocked Shh signaling in HSC-T6 cells, resulting in the decreased protein expression of Patched-1 (PTCH-1), Sonic hedgehog (Shh), and glioma-associated oncogene homolog 1 (GLI1). In short, these results indicate that HZ-6h inhibits liver fibrosis by downregulating MeCP2 through the Shh pathway in TGF-ß1-induced HSC-T6 cells.

NLRC5 regulates cell proliferation, migration and invasion in hepatocellular carcinoma by targeting the Wnt/ß-catenin signaling pathway.

NLRC5, the largest member of nucleotide-binding and oligomerization domain (NOD)-like receptor (NLR) family, has been reported to regulate immune responses and is associated with chronic inflammatory diseases. However, the biological function of NLRC5 in hepatocellular carcinoma (HCC) has not yet been well demonstrated. In this study, the role of NLRC5 in hepatocellular carcinoma cell proliferation, migration and invasion capacities was evaluated by using MTT, flow cytometry, wound healing, transwell assay, and tumor formation assay in nude mice. Western blot analysis and qPCR assay were performed to assess NLRC5 interacting with the activation of Wnt/ß-catenin signaling pathway. Here, we demonstrate that NLRC5 was highly expressed in HCC. Knockdown of NLRC5 significantly inhibited cell proliferation, migration, invasion and the tumor formation in nude mice, and arrested the cell cycle at G0/G1 phase. Furthermore, overexpression of NLRC5 promoted the proliferation, migration and invasion of HCC cells in vitro. Interestingly, we found that up-regulation of NLRC5 not only positively correlates with the increase of ß-catenin but also coordinates the activation of downstream Wnt/ß-catenin signaling pathway. Thus, our findings suggest that NLRC5 may play an important role in progression of HCC and provide a potential therapeutic value in this tumor.

Inhibition of IRF3 expression reduces TGF-ß1-induced proliferation of hepatic stellate cells.

Therapeutic management of liver fibrosis remains an unresolved clinical problem. Activation of hepatic stellate cell (HSC) is a pivotal event in the progression of liver fibrosis. Recent reports have showed that inhibition of activated HSC proliferation contributes to the reversal of liver fibrosis. Interferon regulatory factor 3 (IRF3), one member of the interferon regulatory factor (IRF) family, is recently proven to be a critical modulator in cardiac fibrosis. And accumulating evidence demonstrated that IRF3 plays a crucial role in liver diseases, such as hepatic steatosis, liver inflammation, and alcoholic liver injury. However, the understanding of the function of IRF3 in liver fibrosis remains limited. Our results identified the role of IRF3 in regulating human HSC (LX-2 cell) cell proliferation and apoptosis. The present study indicated that the expression of IRF3 was significantly increased in HSCs in response to TGF-ß1 stimulation. Moreover, a stable and unlimited source of human HSC, the LX-2 cell line, transfected with IRF3-siRNA significantly decreases the expression level of type I collagen (Col1a1) and α-smooth muscle actin (α-SMA) in activated LX-2 cells. On the contrary, overexpression of IRF3 gives rise to an upregulation of Col1a1 and α-SMA in LX-2 cells, and further promoted HSC proliferation. Moreover, the inhibition of IRF3 significantly suppressed TGF-ß1-induced HSC proliferation and increased its apoptosis. Of note, the present study indicated IRF3 may regulate LX-2 cell proliferation by via AKT signaling pathway. In summary, these observations suggest IRF3 may function as a novel regulator to modulate TGF-ß1-induced LX-2 proliferation, at least in part, via AKT signaling pathway.

NLRC5 Mediates IL-6 and IL-1ß Secretion in LX-2 Cells and Modulated by the NF-κB/Smad3 Pathway.

Recent data have shown that nucleotide-binding domain leucine-rich repeat proteins (NLRs), a class of innate immune receptors that respond to pathogen attack or cellular stress, have gained increasing attention. NLRC5 (NLR family, CARD domain containing 5) is the largest member of the NLR family, which has recently been identified as a critical regulator of immune responses. Until recently, the function of NLRC5 has been a matter of debate. In this study, we explore the role of NLRC5 in cytokine secretion and the role of the nuclear factor-κB (NF-κB) signaling pathway in tumor necrosis factor-alpha (TNF-α)-induced NLRC5 expression in LX-2 cells. We demonstrated that overexpression of NLRC5 results in an upregulation of IL-6 and IL-1ß secretion. On the other hand, knockdown of NLRC5 by transfecting siRNA decreased IL-6 and IL-1ß secretion in LX-2 cells. Meanwhile, the results showed that pyrrolidine dithiocarbamate (PDTC) (a specific inhibitor of the NF-κB signaling pathway) inhibited NLRC5 expression and NLRC5 silencing could increase the expression levels of p65 in cell nucleus accompanied with upregulated phosphorylation of Smad3 protein levels in response to TNF-α. These results indicated that NLRC5 plays a significant role in TNF-α-enhanced cytokine (IL-6 and IL-1ß) secretion of LX-2 cells and the NF-κB/Smad3 signal pathway is involved in its induction of expression.

[Correlation of the methylation status of CpG islands in the promoter region of 10 genes with the 5-Fu chemosensitivity in 3 breast cancer cell lines].

OBJECTIVE: To explore the relationship between the methylation status of CpG islands in the promoter region of 10 genes in breast cancer cells and their sensitivity to 5-fluouracil (5-Fu), and to identify the genes responsible for the 5-Fu resistance in breast cancer. METHODS: Three cell lines (differently resistant to chemotherapy) were used in this study: Bcap-37 (IC(50): 289.77 microg/ml), T47D (IC(50): 134.16 microg/ml) and ZR-75-30 (IC(50): 4.20 microg/ml). The methylation profile of 10 genes (BAG1, C11ORF31, CBR1, CBR4, GJA1, FOXL2, IGFBP6, P4HA1, SRI and TYMS) in the 3 breast cancer cell lines was determined by methylation specific PCR. The steady-state mRNAs of ABCC8, CHFR and IGFBP6 genes were quantified by real-time RT PCR analysis. RESULTS: Among the 10 genes, only genes IGFBP6 and FOXL2 displayed differential DNA methylation pattern between the 5-Fu-resistant and 5-Fu-sensitive cell lines. The mRNA expression level of genes PRSS21, LOX, IGFBP6, ABCC8 and CHFR was quantified by real-time RT-PCR analysis. Except for CHFR, the expression level of the other 4 genes was correlated with the methylation status of CpG islands, namely, a lower expression level with methylation status and a higher level with demethylation status. CONCLUSION: The results of the present study have demonstrated that there are 8 genes with differential methylation status in chemosensitive and chemoresistant breast cancer cell lines, i.e. two genes more than the six genes we reported previously. Our findings provide both mechanistic insights for the drug resistance of breast cancer and the basis for further studies on potential application of the DNA methylation in this set of genes for prediction of chemosensitivity of breast cancer.