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.

The level of mercury contamination in mariculture sites at the estuary of Pearl River and the potential health risk.

In the present study, the Hg contamination in mariculture sites located at the estuary of Pearl River was to investigate with an attempt to analyse associated health risks of dietary exposure to both total mercury (THg) and methyl mercury (MeHg) in cultured fish and shellfish. The highest total mercury concentration (7.037 ± 0.556 ng L-1) of seawater was observed at Zhuhai Estuary. The Hg concentrations of sediment in Guishan Island were significantly higher (p < 0.05) than in Daya Bay (away from the Pearl River). Besides, the both THg and MeHg levels in sediment at mariculture sites were higher (p < 0.05) than corresponding reference sites. It was attributed to the fact that mariculture activities increased Hg loading and promoted MeHg production. The vertical distribution of Hg in sediment cores demonstrated that mercury methylation mostly occurred at the sediment-water interface. Results of health risk assessments showed that fish consumption would impose a higher risk to children but less to adults, while shellfish produced in the studied area was safe for consumption.

Conservation and population genetic diversity of Curcuma wenyujin (Zingiberaceae), a multifunctional medicinal herb.

Curcuma wenyujin is an important multifunctional medicinal herb in China. Currently, populations of C. wenyujin are decreasing, and wild individuals have almost disappeared from their natural habitats. Moreover, little is known regarding the molecular characteristics of this plant. In this study, we investigated the genetic diversity and variation of five populations of C. wenyujin, using ran-dom amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers. We found that the percentages of polymorphic loci (PPL) at the species level (98.25% by RAPD and 100% by ISSR) were significantly higher than those at the population level (66.32% by RAPD and 67.14% by ISSR). The highest values of PPL, expected heterozygosity, and Shannon's information index were in Pop1, while the lowest values were in Pop2. Both DNA markers revealed a short genetic distance between Pop1 and Pop2 (0.1424 by RAPD and 0.1904 by ISSR). Phylogenetic trees produced similar results, with Pop1, Pop2, and Pop5 in one group and Pop3 and Pop4 in another. There were no significant correlations between their genetic distances and their geographical distances. The highest genetic diversity was in Pop1 and the lowest was in Pop2, and genetic diversity at the species level was relatively low, but much higher than that at the population level. We recommended the establishment of a germplasm bank, in situ con-servation, and propagation of wild individuals. The present study will improve the evaluation, protection, and utilization of the population resources of C. wenyujin.

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.

An esophagogastroduodenal anastomosis model for esophageal adenocarcinogenesis in rats and enhancement by iron overload.

The aim of this study is to establish a good animal model for esophageal adenocarcinoma (EAC) and to test the hypothesis that iron over-nutrition enhances EAC formation. With rats, esophagogastroduodenal anastomosis (EGDA) was accomplished by anastomosing the duodenum to the gastroesophageal junction. Iron supplementation was given by i.p. injection of iron dextran (4 mg Fe/kg/week). This model mimics the development of human EAC by introducing mixed reflux of gastric and duodenal contents. At 40 weeks after surgery, the body weight, food intake, hemoglobin, total serum iron, transferrin saturation, serum albumin, and plasma levels of alpha-tocopherol, gamma-tocopherol and retinol of the EGDA rats were not significantly different from those of the non-operated controls. The animals generally had only mild esophagitis, except that the area surrounding the anastomosis opening had more severe esophagitis. Columnar-lined esophagus (CLE), CLE with dysplasia, and EAC were diagnosed in 53.5, 34.9 and 25.6%, respectively, of the 43 rats. Intraperitoneal iron supplementation significantly enhanced esophageal lesions with CLE, CLE with dysplasia, and EAC to 78.0, 53. 7 and 53.7%, respectively, of the 41 rats. All the tumors were well-differentiated mucinous adenocarcinomas at the squamocolumnar junction area, where most iron deposition was observed. EGDA avoids nutritional problems seen in other animal models for EAC. We believe that direct anastomosis of squamous epithelium to columnar epithelium and mixed reflux of gastric and duodenal contents lead to the formation of CLE and EAC. With this model, we demonstrated that iron supplementation significantly enhanced EAC formation, suggesting that iron over-nutrition could also be a risk factor for human EAC.