Necrosulfonamide (NSA) protects intervertebral disc degeneration via necroptosis and apoptosis inhibition.

OBJECTIVE: Previous studies have shown that nucleus pulposus (NP) cell death plays an extremely important role in the progress of intervertebral disc degeneration (IVDD). This research aimed to investigate the protective effect of the MLKL inhibitor necrosulfonamide (NSA) on human NP cells. PATIENTS AND METHODS: We collected human NP tissues from the patients undergoing disc herniation operations and isolated NP cell from the samples. IL-1ß (10 ng/ml) was used to establish a NP cells degenerated model. We analyzed the expression of caspase 3, caspase 8, RIPK1, RIPK 3, and MLKL in different degree of degenerate disc tissues. Cell viability was analyzed by the Cell Counting Kit-8 (CCK-8) assay. The expression levels of collagen Ⅱ, ß-galactosidase (ß-gal), caspase 3, caspase 8, RIPK1, RIPK 3, and MLKL, several inflammatory and anti-oxidant enzymes of different NP cell treat groups were detected by Western blotting, immunofluorescence staining, or RT-PCR. Flow cytometry was used to measure the ROS level and cell apoptosis. RESULTS: The data showed that expression of caspase 3, caspase 8, RIPK1, RIPK 3, and MLKL markedly increased in severely degenerated disc tissues. IL-1ß promoted the cell death of NP cells, while NSA could reverse the effects of IL-1ß. We found that NAS increased the antioxidant SOD1, SOD2, CAT, and GPX3 expression and suppressed oxidative stress in the disc. Moreover, MMP3, MMP10, IL-6, and TNF-α were significantly suppressed by the NSA. CONCLUSIONS: These results suggest that NSA prevented NP degradation via inhibiting apoptosis and necroptosis of NP cells. Besides, the protective function of antagonizing cell death may owe to the inflammation and oxidative stress suppression.

Evaluation of the application of high volume hemofiltration in sepsis combined with acute kidney injury.

OBJECTIVE: To investigate the safety and effectiveness of the application of high-volume hemofiltration (HVHF) in children with sepsis combined with acute kidney injury. PATIENTS AND METHODS: A total of 76 child patients were enrolled and randomly divided equally (n=38) into control and the observation groups respectively. The control group received conventional volume hemofiltration (the ultrafiltration rate of 35-50 ml/kg/h), and the observation group received HVHF (50-100 ml/kg/h). RESULTS: The serum Interleukin-6 (IL-6), Tumor Necrosis Factor-a (TNF-α) and creatinine levels were significantly lower in the observation group than the control group at 6 h, 12 h, 24 h and 48 h of hemofiltration (p<0.05). The efficacy rate of treatment was improved. The mortality rate and incidence rate of complications were decreased, and the treatment course was significantly shortened (p<0.05). CONCLUSIONS: The application of HVHF in children with sepsis combined with acute kidney injury has a better safety and effectiveness.

Function of TGF-beta and p38 MAKP signaling pathway in osteoblast differentiation from rat adipose-derived stem cells.

BACKGROUND: Adipose-derived stem cells (ADSCs) are multipotent progenitors that can commit to osteoblast, chondrocyte, adipocyte and several other lineages. The proper utilization of stem cells for clinical application requires an integrated understanding of multiple signal inputs that control maintenance of stemness, proliferation and differentiation. MATERIALS AND METHODS: In this study, ADSCs in rat were isolated from subcutaneous tissues of abdomen and inguinal fat pads, purified and expanded in vitro. Bone morphogenetic protein 2, TGF-beta 1, SB203580 (P38 MAPK inhibitor), Noggin (BMP inhibitor) and SB431542 (TGF-beta inhibitor) were used for differentiation into osteoblasts. RESULTS: Both TGF-beta signaling pathway and p38 MAKP signaling pathway could affect the differential direction of the ADSCs. PCR assays indicated that both TGF-beta signaling pathway and p38 MAKP signaling pathway played a crucial roles in osteoblasts differentiation of the ADSCs, the members included Smad 1, Smad 5, Smad 8, P38, ASK1, MKK3, MKK6, Runx 2, collagen type 1, and osteopontin. CONCLUSIONS: This research provides a theoretical basis and experimental evidence for therapeutic application of rat ADSCs to treat bone injury.