Circular RNA circ-FAM158A promotes retinoblastoma progression by regulating miR-138-5p/SLC7A5 axis.

BACKGROUND: Mounting evidence has shown that circular RNAs (circRNAs) have vital roles in human cancers, including retinoblastoma (RB). The purpose of this study was to investigate the exact roles and underlying mechanism of circRNA ER membrane protein complex subunit 9 (circ-FAM158A) in RB. METHODS: Quantitative real-time polymerase chain reaction (qRT-PCR) was carried out to determine the expression levels of circ-FAM158A, miR-138-5p and solute carrier family 7 member 5 (SLC7A5). Cell proliferation was evaluated by Cell counting Kit-8 (CCK-8) assay and colony formation assay. Flow cytometry analysis was applied to determine cell cycle distribution and apoptosis rate. Transwell assay was conducted to assess cell migration and invasion. The interaction between miR-138-5p and circ-FAM158A or SLC7A5 was predicted by starBase v2.0 and confirmed by dual-luciferase reporter assay. Western blot assay was performed to examine the protein expression of SLC7A5. The mice xenograft model was established, immunohistochemistry (IHC) and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling (TUNEL) assays were conducted to confirm the role of circ-FAM158A in vivo. RESULTS: Circ-FAM158A and SLC7A5 were overexpressed and miR-138-5p was downregulated in RB tissues and cells. Circ-FAM158A knockdown inhibited RB cell proliferation, metastasis, and promoted apoptosis in vitro and in vivo. MiR-138-5p was a direct target of circ-FAM158A, and miR-138-5p inhibition reversed the inhibitory effect of circ-FAM158A silence on the progression of RB cells. Additionally, SLC7A5 was identified as a target of miR-138-5p, and SLC7A5 overexpression abated the anti-tumor roles of miR-138-5p in RB cells. Besides, circ-FAM158A positively regulated SLC7A5 expression by sponging miR-138-5p. CONCLUSION: Circ-FAM158A knockdown inhibited the progression of RB by regulating miR-138-5p/SLC7A5 axis, which provided new insights into the pathogenesis of RB.

Producing and storing self-sustaining drinking water from rainwater for emergency response on isolated island.

Drinking water on isolated islands includes treated rainwater, water shipped from the mainland, and desalinated seawater. However, marine transportation and desalination plants are vulnerable to emergencies, such as extreme weather, making self-sustaining stand-by water for emergency response essential. Rainwater is ideal for producing the stand-by water, and rainwater harvesting is sustainable and clean, and prolonged biostability can be ensured by managing biological and chemical parameters. The present study applied a stand-by drinking water purification system (primarily including nanofiltration and low-dose chlorination) to explore the feasibility of producing and storing cleaner drinking water from rainwater and the following conclusions were drawn. First, treatment of rainwaters ensures biosafety for seven days, which is longer than that for untreated rainwater; the proportion of opportunistic pathogens decreased from 23.40-7.77% after nanofiltration, and it was proposed that the microbial community converges after advanced water treatment. Second, chemical qualities were improved. Local resource coral sand prevents pH in rainwater from decreasing below 6.5, and treated rainwater had lower disinfection by-product potential and higher disinfection efficiency, allowing periodical rainwater recycling. Third, harvesting rainwater was extremely cost-effective, with an operation cost of 1.5-2.5 RMB/m3. From biosafety, chemical safety, and economic cost perspectives, self-sustaining water from rainwater can contributes to the development of sustainable and cost-effective water supply systems on isolated islands. Mixing treated rainwater and desalinated seawater reasonably guarantees sufficiency and safety.

Long non-coding RNA colon cancer-associated transcript 2 may promote esophageal cancer growth and metastasis by regulating the Wnt signaling pathway.

The aim of this study was to investigate how long non-coding (lnc)RNA colon cancer-associated transcript 2 (CCAT2) regulates the proliferation, invasion and metastasis of esophageal cancer cells via the Wnt signaling pathway. The expression of lncRNA CCAT2 was quantified by reverse transcription-quantitative PCR in four esophageal cancer cell lines (Eca-109, EC9706, KYSE150 and TE-1) and normal human esophageal epithelial cells (HEECs). The effect of silencing CCAT2 (si-CCAT2) and inhibiting Wnt signaling (using the inhibitor FH535) on the proliferation, migration and invasion of Eca-109 cells was measured by MTT, wound-healing and Transwell invasion assays. Flow cytometry was used to evaluate apoptosis in si-CCAT2 Eca-109 cells. The expression of ß-catenin and proliferating cell nuclear antigen (PCNA) proteins was detected by immunohistochemistry. The pro-apoptotic protein Bax, cyclin D1 and Wnt target proteins, including c-Myc and adenomatous polyposis coli (APC), were detected by western blotting. LncRNA CCAT2 was highly expressed in the four esophageal cancer cell lines compared with the HEEC cells. The expression of CCAT2 was significantly decreased in si-CCAT2 Eca-109 cells. Treatment with si-CCAT2 and FH535 alone or in combination significantly inhibited the proliferation, migration and invasion of Eca-109 cells. The treatments also promoted apoptosis, upregulated the expression of Bax and APC proteins, and downregulated ß-catenin, PCNA, cyclin D1 and c-Myc proteins. In summary, lncRNA CCAT2 is upregulated in esophageal cancer cells and the knockdown of lncRNA CCAT2 inhibits their proliferation, migration and invasion via the Wnt signaling pathway.

Capsaicin induces apoptosis and autophagy in human melanoma cells.

Melanoma is the most lethal type of skin cancer; rapid metastasis and resistance to conventional radio- and chemotherapy make melanoma the most aggressive type of skin cancer. In addition, there is a high recurrence rate within 1 year among patients with melanoma following traditional treatment by chemotherapy or immunotherapy, and these treatment options are only useful in advanced stages. As the efficiency of treatment options for melanoma is not ideal, the present study aimed to confirm that capsaicin has inhibitory effects on the human melanoma A375 and C8161 cell lines in vitro. Capsaicin, the active component of peppers, has been reported to possess substantial anticarcinogenic and antimutagenic activities. Additionally, capsaicin exhibits an inhibitory effect on tumor growth in numerous malignant cell lines. In the present study, flow cytometry, fluorescent puncta detection and western blotting were performed. The experimental results indicated that capsaicin activated apoptosis, and that apoptosis induction was associated with poly(ADP-ribose) polymerase (PARP) cleavage and activation of caspase-3. Additionally, the formation of autophagosomes and accumulation of microtubule-associated proteins 1A/1B light chain 3B-II and beclin 1 suggested that capsaicin induced autophagy in human melanoma cells. Furthermore, inhibiting capsaicin-induced autophagy promoted the activation of cleaved caspase-3 and PARP proteins, which are associated with apoptosis. In addition, inhibition of autophagy using 3-MA enhanced capsaicin-induced cell death, indicating that capsaicin-induced autophagy is a pro-survival process in cells. In conclusion, the results of the present study revealed that capsaicin induced cell apoptosis and autophagy in human melanoma cells and capsaicin may be considered as a novel candidate drug for melanoma treatment.

Kaempferol attenuates retinal ganglion cell death by suppressing NLRP1/NLRP3 inflammasomes and caspase-8 via JNK and NF-κB pathways in acute glaucoma.

AIMS OR PURPOSE: Glaucoma is the leading cause of vision loss and blindness in the world. Elucidating the pathogenesis of glaucoma and developing effective treatment should be the priority. Inflammation and oxidative stress play essential roles in glaucoma pathogeneisis. Kaempferol is a natural flavonol and has anti-inflammatory and anti-oxidative activities. In this study, we explored the potential effects of kaempferol on acute glaucoma. METHODS: We established the retinal ischemia-reperfusion (I/R) mice model and administrated kaempferol to I/R mice. We monitored the retina thickness change, retinal ganglion cell (RGC) death, caspase-8 and caspase-3 activation, NLRP1/NLRP3 inflammasomes activation, pro-inflammatory cytokines production, and activations of NF-κB and MAPKs signaling pathways. RESULTS: Kaempferol prevented retina thickness change and RGC death in I/R mice. The activations of caspase-8, caspase-3, and NLRP1/NLRP3 inflammasome activation were inhibited by kaempferol. Kaempferol prevented pro-inflammatory cytokines productions in I/R mice. The activation of NF-κB and JNK signaling pathways was also inhibited by Kaempferol in I/R mice. CONCLUSION: Kaempferol attenuated retinal ganglion cell death by suppressing NLRP1/NLRP3 inflammasomes and caspase-8 via inhibiting NF-κB and JNK pathways in acute glaucoma.

Effects of transplanted adipose derived stem cells on the expressions of α-SMA and DCN in fibroblasts of hypertrophic scar tissues in rabbit ears.

To study the effects of transplanted adipose derived stem cells (ADSCs) on the expressions of α-smooth muscle actin (α-SMA) and decorin (DCN) in fibroblasts of hypertrophic scar tissues in rabbit ears. Twelve New Zealand white rabbits were selected; the normal subcutaneous adipose tissues in inguinal region were removed, ADSCs were extracted via enzyme digestion, cultured in Dulbecco's modified Eagle's medium (DMEM) and inoculated into the culture dish (3-5×104 cells/ml). After the rabbit ear hypertrophic scar model was established successfully, the fibroblasts of hypertrophic scar tissues in rabbit ears were separated and cultured using the mechanical method combined with enzyme digestion, and the ADSCs and scar fibroblasts were cultured in non-contact Transwell co-culture system for 21 days (experimental group); the corresponding scar fibroblasts were cultured in an ordinary 6-well plate without any treatment for 21 days (control group). The content of collagen I in fibroblasts was detected using the enzyme-linked immunosorbent assay (ELISA) kit, the mRNA expressions of α-SMA and DCN were detected via reverse transcription-polymerase chain reaction (RT-PCR), the protein expressions of α-SMA and DCN were detected via western blot analysis, and the expressions and distribution of α-SMA and DCN were detected via immunofluorescence assay. The results of ELISA showed that the content of collagen I in experimental group was decreased significantly (p<0.01). The results of RT-PCR and western blot analysis revealed that the mRNA and protein expression levels of α-SMA were significantly decreased (P<0.01, but those of DCN were significantly increased (p<0.01). Moreover, the results of immunofluorescence assay showed that the expression of α-SMA in experimental group was significantly decreased, while the expression of DCN was significantly increased. ADSCs can inhibit the mRNA and protein expressions of α-SMA and promote the mRNA and protein expressions of DCN in in vitro culture system, and they are expected to be used in the prevention and treatment of pathological scars.

Resveratrol alleviates LPS-induced injury in human keratinocyte cell line HaCaT by up-regulation of miR-17.

BACKGROUND: Resveratrol (RSV), an edible polyphenolic phytoalexin, plays an important role in ameliorating inflammation, including skin inflammation after burn injury. However, the specific molecular mechanism underlying its anti-inflammation effect is still unclear. Herein, the effect and the mechanism underlying the protection of HaCaT cells by RSV against inflammation were examined. METHODS: Lipopolysaccharide (LPS)-induced inflammation and the cytoprotection of RSV were evaluated by detecting viability, apoptosis, expressions of apoptosis-associated proteins and the productions of pro-inflammatory factors by CCK-8 assay, flow cytometer, Western blot, and qRT-PCR. miR-17 expression in RSV-treated HaCaT cells was determined by qRT-PCR. The role of miR-17 in protective effect of RSV was investigated after altering its expression using transfection assay. The main ingredients in PTEN/PI3K/AKT and mTOR pathways were quantified by Western blot. RESULTS: LPS-induced HaCaT cell injury was inhibited by RSV administration. RSV promoted viability, inhibited apoptotic cell rate, increased Bcl-2 expression, decreased Bax, cleaved-Caspase-3, and cleaved-Caspase-9 expressions. RSV also inhibited inflammation injury of HaCaT cells by reducing productions of IL-6, IL-8, and TNF-α. miR-17 was up-regulated in LPS and RSV-co-treated cells. The protective effect of RSV might contribute to overexpression of miR-17. In the mechanism study, RSV-miR-17 axis was found to activate PTEN/PI3K/AKT and mTOR pathways in LPS-treated cells. CONCLUSION: RSV alleviated LPS-induced injury in human keratinocyte cell line HaCaT through activations of PTEN/PI3K/AKT and mTOR pathways, which were modulated by miR-17.

Combination of autologous transplantation of G-CSF-mobilized peripheral blood mononuclear cells and Panax notoginseng saponins in the treatment of unreconstructable critical limb ischemia.

BACKGROUND: The aim of this study is to explore the efficacy and safety of the combination of autologous transplantation of granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood mononuclear cells (PBMNCs) and Panax notoginseng saponins (PNS) in the treatment of unreconstructable critical limb ischemia (CLI). METHODS: We performed an open-label, parallel-group, single-center, randomized clinical trial in this study. A total of 52 patients were enrolled and randomly divided into 2 groups (the PBMNC + PNS group and the PBMNC group) in a 1:1 ratio. Evaluation variables, including changes in the ankle-brachial index (ABI) of ischemic limbs, ulcer area, severity of rest pain, transcutaneous oxygen pressure (T(C)PO2), and 6-min walk distance from baseline to week 8 and 16, as well as angiographic scores for new collateral vessel formation at week 16, were used to compare the benefits of these 2 treatment approaches. RESULTS: After 16 weeks of treatment, improvement in ABI, T(C)PO2, and 6-min walk distance was significantly better in the PBMNC + PNS group. In addition, the combination of PBMNC transplantation and PNS administration yielded a greater reduction in ulcer area and severity of rest pain than did PBMNC transplantation alone. The proportion of patients experiencing any adverse event was similar between both treatment groups. Adverse events caused by PBMNC transplantation or PNS were generally mild and no serious adverse events occurred throughout the entire period of study. CONCLUSIONS: A combination of PNS and PBMNC transplantation appears to be a safe and effective treatment for patients with unreconstructable CLI. This combination may have great potential advantages in comparison with PBMNC transplantation alone and might constitute a novel therapeutic option for unreconstructable CLI.