LncSNHG14 promotes the development and progression of bladder cancer by targeting miRNA-150-5p.

OBJECTIVE: To elucidate whether lncSNHG14 could influence the proliferative potential and cell cycle progression of bladder cancer cells via binding to microRNA-150-5p (miRNA-150-5p). We aim to investigate the potential mechanism of miRNA-150-5p in the occurrence and progression of bladder cancer (BCa). PATIENTS AND METHODS: Expression levels of SNHG14 and miRNA-150-5p in BCa tissues and normal bladder tissues were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Their expressions in BCa cell lines were detected as well. Regulatory effects of NHG14 and miRNA-150-5p on proliferative potential and cell cycle progression were evaluated by cell counting kit-8 (CCK-8) and flow cytometry, respectively. Through the dual-luciferase reporter gene assay, binding conditions between SNHG14 and miRNA-150-5p, as well as between miRNA-150-5p and synaptic vesicle-associated membrane protein 2 (VAMP2), were verified. Finally, rescue experiments were performed to clarify whether SNHG14 regulated behaviors of BCa cells by absorbing miRNA-150-5p to degrade VAMP2. RESULTS: SNHG14 was highly expressed in BCa tissues and cell lines. The overexpression of SNHG14 accelerated the proliferative potential and cell cycle progression of BCa cells. SNHG14 was confirmed to bind to miRNA-150-5p. MiRNA-150-5p remained a low expression in BCa tissues. Moreover, miRNA-150-5p overexpression suppressed proliferative potential and cell cycle progression of BCa cells, which could reverse the promotive role of SNHG14 on behaviors of BCa cells. Furthermore, VAMP2 was the target gene of miRNA-150-5p. VAMP2 overexpression reversed the biological function of miRNA-150-5p in inhibiting proliferative potential and cell cycle progression of T24 and UC9 cells. CONCLUSIONS: LncSNHG14 overexpression accelerates proliferative potential and cell cycle progression of BCa cells through absorbing miRNA-150-5p to degrade VAMP2 expression.

Effects of siRNA-mediated silencing of Sal-like 4 expression on proliferation and apoptosis of prostate cancer C4-2 cells.

The aim of this study was to evaluate the effects of small interfering RNA (siRNA)-inhibited expression of the Sal-like 4 (SALL4) gene on the proliferation, colony formation, and apoptosis of prostate cancer C4-2 cells. C4-2 cells were cultured and divided into a si-SALL4 group, a negative control siRNA group, and a blank control group. SALL4 mRNA levels and protein expression were detected by real-time polymerase chain reaction and western blot, respectively. Changes in the cell proliferation and colony formation capacities were observed by using the MTS colorimetric method and colony formation assay, respectively. The influence of SALL4 on apoptosis was assessed with flow cytometry, and the expression of apoptosis-related proteins B-cell lymphoma 2 (Bcl-2) and bcl-like-protein 4 (Bax) were detected by western blot. The si-SALL4 group had significantly lower mRNA and protein levels of SALL4 as well as decreased proliferation and colony formation capacities than the negative control group (P < 0.05). There were significantly more apoptotic cells in the si-SALL4 group compared to the negative control (P < 0.05), and the expression of Bcl-2 and Bax decreased and increased, respectively, after treatment with si-SALL4. Silencing SALL4 expression by using siRNA technology inhibited the proliferation and colony formation of C4-2 cells, and promoted apoptosis likely mediated by Bcl-2 and Bax expression. These results provide experimental basis for further elucidating the role of SALL4 in prostate cancer cells.

In vivo determination of muscle-derived stem cells in rat corpus cavernosum.

The aim of this in vivo study was to determine the existence of muscle-derived stem cells (MDSCs) in rat corpus cavernosum. Immunohistochemical and RT-PCR analyses were performed to determine the expression of the stem cell markers (Sca-1, Oct4, and desmin) in Sprague-Dawley (SD) rats in different age groups (10 rats in each group). Sca-1 was mainly expressed in blood vessels and cavernous sinus and demonstrated primarily cytoplasmic staining. Desmin was expressed mainly in muscle tissues and staining occurred mainly in the cytoplasm but also partially in the nucleus. An extremely small amount of double-positive stained cells (Sca-1/desmin) were detected near the cavernous sinus. Expression of the markers was significantly and negatively correlated with the age of the rats (P < 0.05). The RT-PCR results showed that the expression levels of Sca-1 and desmin significantly decreased with age (P < 0.05). Correlation analysis indicated that the expression of Sca-1 and desmin were significantly and negatively correlated with the age of rats (r = -0.929, P < 0.05). The present study provides evidence for the existence of MDSCs in rat corpus cavernosum. MDSCs may have therapeutic potential in the treatment of organic erectile dysfunction.

Advanced glycation end products upregulate C-reactive protein synthesis by human hepatocytes through stimulation of monocyte IL-6 and IL-1 beta production.

Patients with chronic renal failure are characterized by increased plasma levels of C-reactive protein (CRP) and advanced glycation end products (AGE). AGE have been identified as a class of proinflammator mediators. To investigate whether AGE can stimulate hepatocytes to produce CRP, primary human fetal hepatocytes (HFH) were incubated with AGE-modified human serum albumin (AGE-HSA) or conditioned medium from AGE-HSA-stimulated monocytes (AGE-MCM). CRP concentrations in the supernatants were determined by an ELISA and CRP mRNA levels were determined by a quantitative RT-PCR. Exposure of HFH with AGE-HSA for 12-72 h did not change CRP concentrations in the supernatants. CRP protein and mRNA expression were significantly upregulated in a time- and dose-dependent manner when HFH were incubated with AGE-MCM. This stimulating effect was partially inhibited when AGE-MCM were preincubated with antibodies against interleukin-6 (anti-IL-6), interleukin-1 beta (anti-IL-1 beta), or soluble IL-1 receptor and was completely inhibited when AGE-MCM were preincubated with anti-IL-6 and anti-IL-1 beta simultaneously. The inhibiting effect did not occur when AGE-MCM was preincubated with antibody of tumour necrosis factor-alpha (anti-TNF-alpha) and soluble TNF receptor. Exposure of HFH with exogenous IL-6 and IL-1 beta, at the same concentrations as contained in AGE-MCM, also increased CRP production, but exogenous TNF-alpha had no effect. These results suggest that AGE cannot directly stimulate hepatocytes to produce CRP, but rather indirectly enhance CRP expression via stimulation of IL-6 and IL-1 beta production by human monocytes.

[The primary study on rabbit bone marrow mesenchymal stem cells in constructing tissue-engineered cartilage].

OBJECTIVE: To study the feasibility of constructing tissue engineered cartilage by differentiated rabbit bone marrow mesenchymal stem cells(MSC) cultured in vitro and in vivo. METHODS: The MSC were isolated from the nucleated cells fraction of autologous bone marrow by density gradient centrifuge, and then induced into chondrogenic differentiation by adding dexamethasone, transforming growth factor-beta 1 (TGF-beta 1) and ascorbic acid in vitro. After 3 weeks, some cells turned to round shape and secreted metachromatic matrix. The cartilaginoid grafts composed of chondrogenic MSC. Bovine type I collagen and human fibrin were cultured within the chondrogenic medium for 2 weeks in vitro or transplanted subcutaneously adjacent to the knee joint for 3 weeks in vivo. RESULTS: The most cells in the grafts were degenerated and disappeared after cultured in vitro. But the residual cells were survival and secreted metachromatic staining proteoglycan with toluidine blue, which was characteristic cartilage matrix. The grafts developed into matured cartilage tissue assessed by histological examination after 3 weeks of transplantation in vivo. CONCLUSION: MSC can be used as functional cells to constructing tissue engineered cartilage.

[Diagnosis and treatment of adrenal cysts].

Six patients with adrenal cysts, a rare disease, were treated. Among 5 patients undergoing operations, 4 had pseudocysts and one had epithelial cyst. Most of the patients had no symptoms. B-ultrasonography and CT scanning played a leading role in the diagnosis of the disease. The incidence, etiology, pathological classification and treatment of adrenal cysts are discussed.