[Effect of Toll-like receptor 2 on the inhibition role of sevoflurane on airway inflammation in asthmatic mice].

OBJECTIVE: To investigate the effect of Toll-like receptor 2 (TLR2) on the inhibition role of sevoflurane on airway inflammation in asthmatic mice. METHODS: The lung tissue samples of C(57) BL/6 mice used in this study were from previous research, including control group, asthma group and sevoflurane group, 8 samples in each group. Twenty-four specific pathogen free female TLR2 gene deletion (TLR2(-/-)) mice were randomly assigned to control group, asthma group and sevoflurane group, with 8 mice in each group. Asthma group and sevoflurane group were then sensitized and challenged with ovalbumin (OVA) to establish asthma model, combined with repeated inhalation of 3% sevoflurane in sevoflurane group. In C(57) mice, expression levels of TLR2 were detected using Western blotting analyses. In TLR2(-/-) mice, numbers of differential inflammatory cells were investigated; levels of tumor necrosis factor-alpha (TNF-α) and interleukin-10 (IL-10) in bronchoalveolar lavage fluid (BALF) were measured by enzyme linked immunosorbent assay (ELISA); lung tissue inflammation was detected with HE staining. RESULTS: In lung tissues from C(57) mice, levels of protein expression of TLR2 in asthma group (0.547±0.042) were higher than those in control group (0.312±0.023) (P=0.023) and sevoflurane group (0.287±0.033) (P=0.020). In TLR2(-/-) mice, the number of total cells ((83.13±19.43)×10(3)/ml), numbers of differential inflammatory cells and TNF-α level ((546±16) pg/ml) in BALF in sevoflurane group were lower than those in asthma group ((206.43±41.82)×10(3)/ml, (732±41) pg/ml), but still higher than those in control group ((44.64±7.17)×10(3)/ml, (380±24) pg/ml) (all P<0.05); lung tissue inflammation was inhibited in sevoflurane group than in asthma group, but still more obvious than that in control group. CONCLUSION: Toll like receptor 2 involved in the anti-inflammatory effect of sevoflurane on asthmatic airway inflammation in mice.

Prognostic characteristics of duodenal gastrointestinal stromal tumours.

BACKGROUND: This study evaluated the clinical characteristics, surgical procedures and prognosis of duodenal gastrointestinal stromal tumours (GISTs). METHODS: Patients with a diagnosis of primary duodenal GIST treated between January 2000 and December 2012 were analysed. Patients with gastric and small intestinal GISTs were chosen as control groups according to the following parameters: age, tumour size, mitotic index and adjuvant imatinib therapy. Operative procedures for patients with duodenal GIST included pancreaticoduodenectomy or limited resection. Disease-free survival (DFS) was calculated using Kaplan-Meier analysis. RESULTS: Some 71 patients with duodenal, 71 with gastric and 70 with small intestinal GISTs were included in the study. DFS of patients with duodenal GIST was shorter than that of patients with gastric GIST (3-year DFS 84 versus 94 per cent; hazard ratio (HR) 3.67, 95 per cent c.i. 1.21 to 11.16; P = 0.014), but was similar to that of patients with small intestinal GIST (3-year DFS 84 versus 81 per cent; HR 0.75, 0.37 to 1.51; P = 0.491). Patients who underwent pancreaticoduodenectomy were older, and had larger tumours and a higher mitotic index than patients who had limited resection. The 3-year DFS was 93 per cent among patients who had limited resection compared with 64 per cent for those who underwent PD (HR 0.18, 0.06 to 0.59; P = 0.001). CONCLUSION: The prognosis of duodenal GISTs is similar to that of small intestinal GISTs.

Silencing of fat-1 transgene expression in sheep may result from hypermethylation of its driven cytomegalovirus (CMV) promoter.

The fat-1 gene was isolated from roundworm Caenorhabditis elegans, and built into pIRES2-EGFP expression vectors driven by cytomegalovirus (CMV) promoter or cytomegalovirus enhancer and chickenß-actin (CAG) promoter. Both CMV- and CAG-driven expression vectors were transfected to sheep fetal fibroblast cells. Positive transfected cells were used as donors for somatic cell nuclear transfer (SCNT) and the cloned embryos were transferred into the oviducts of synchronized recipient sheep. Two lambs derived from CMV vector and three lambs derived from CAG vector developed to term. Although Southern analyses using tissues from the two lambs derived from CMV vectors indicated integration of fat-1 gene into the genome, fat-1 mRNAs were not detected by RT-PCR. However, there was fat-1 expression (detected by RT-PCR) in tissues from transgenic lambs driven by CAG vectors. To investigate potential mechanisms involved in the two transgene models, methylation state of the vector promoters were examined. In CMV-driven transgenics, CMV promoters had almost no methylation in transfected cells and the resultant cloned embryos, whereas high methylations were detected in tissues and organs in transgenic lambs. In the CAG-driven transgenics, there were almost no methylations in transgenic cells and transgenic cloned embryos, and cloned lambs expressed fat-1 mRNA (detected by RT-PCR). Moreover, although SV40 promoters which drove neo/kan marker gene in CMV vectors were highly methylated in tissues from transgenic lambs, they were without methylation in cells and embryos. Therefore, we concluded that highly methylated CMV promoters induced the silence of fat-1 transgene expression in sheep. Furthermore, CAG promoter, but not CMV promoter was suitable for generation of fat-1 transgenic sheep.

Primary pancreatic pleomorphic leiomyosarcoma.

Primary pancreatic leiomyosarcoma is a rare mesenchymal tumour that is believed to arise from the walls of the pancreatic blood vessels or the pancreatic duct. A 56-year-old female was referred with epigastric pain and abdominal mass. Preoperative computed tomography showed a large soft tissue mass in the pancreatic body and tail. Fine needle aspiration biopsy indicated a spindle cell type tumour. The patient received distal pancreatectomy with no adjuvant treatment. Histology revealed a pleomorphic spindle cell neoplasm with an immunoprofile suggestive of smooth muscle origin. The absence of other lesions in the body was consistent with the diagnosis of primary pleomorphic leiomyosarcoma. The patient was well and tumour-free 14 months after surgery. Detailed immunohistochemical analyses are necessary in the diagnosis of this highly malignant tumour. Radical resection offers the only chance of long-term survival.

Increased epithelial and serum expression of macrophage migration inhibitory factor (MIF) in gastric cancer: potential role of MIF in gastric carcinogenesis.

AIMS: Macrophage migration inhibitory factor (MIF) is implicated in tumorigenesis. This study was conducted to determine whether MIF expression is associated with gastric pathology and whether MIF expression is increased in malignant gastric cells in vitro. MATERIALS AND METHODS: Patients with a normal gastric mucosa, Helicobacter pylori infected gastritis, intestinal metaplasia, and gastric adenocarcinoma were included. Immunohistochemistry and enzyme linked immunosorbent assay (ELISA) were used to determine MIF expression in gastric epithelial cells and MIF levels in serum, respectively. Five gastric cancer cell lines (AGS, MKN-45, MKN-28, MGC-803, and SGC-7901) and one non-malignant gastric cell line (GES-1) were cultured for 24 hours. MIF protein in the supernatant and MIF mRNA in cultured cells were measured by ELISA and reverse transcription-polymerase chain reaction, respectively. RESULTS: The percentage of MIF expressing epithelial cells was low in normal mucosa (12%) but substantially higher in gastritis (52%), intestinal metaplasia (66%), and gastric cancer (96%) (p<0.001, ANOVA). Serum MIF levels were low in patients with a normal mucosa (576 (82) pg/ml) but higher in patients with gastritis (2100 (349) pg/ml), intestinal metaplasia (4498 (253) pg/ml), and gastric cancer (9737 (1249) pg/ml) (p<0.001, ANOVA). There was a correlation between epithelial MIF expression and serum MIF levels (r = 0.776, p<0.001). In vitro, expression of MIF protein and mRNA was increased in malignant cells compared with non-malignant cells. CONCLUSIONS: Epithelial and serum MIF expression was progressively increased in H pylori induced gastritis, intestinal metaplasia, and gastric cancer, suggesting that MIF is involved in gastric carcinogenesis and may be a valuable biomarker for the early detection of gastric cancer.

Effects of calcitriol and its analogues on interaction of MCP-1 and monocyte derived dendritic cells in vitro.

AIM: To study the effect of calcitriol [1,25(OH)2D3] and its analogues on the interaction of monocyte chemoattractant protein-1 (MCP-1) and in vitro generated monocyte-derived dendritic cells (MoDC). METHODS: MoDC were obtained by differentiating monocytes in exposure to GM-CSF and IL-4 for 5 d. mRNA expression of MCP-1 and its receptors were analyzed by RT-PCR, and protein production of MCP-1 by ELISA and migratory ability of MoDC in response to MCP-1 by a micromultiwell chemotaxis chamber assay. RESULTS: MoDC can express MCP-1 mRNA, and secret a low level of MCP-1 protein and has the ability to migrate to MCP-1 in corresponding to its expression of MCP-1 receptors. Calcitriol and its analogues with the same affinity to vitamin D receptor up-regulated the gene expression of both MCP-1 and its receptors, enhanced MCP-1 protein production and promoted the migratory ability of MoDC to MCP-1. CONCLUSION: The interaction of DC and MCP-1 found in this study may suggest a possible auto-regulatory role between DC and MCP-1 and the modulatory effect of calcitriol and its analogues on DC and MCP-1 might provide an understanding of their positive role in tumors.