Periplocoside P (PSP) isolated from the root bark of Periploca sepium contains a pregnane glycoside skeleton and possesses high insecticidal properties. Preliminary studies indicated that PSP disrupts epithelial functions in the midgut of lepidopteran larvae. In the present study, we examined the effects of PSP on the apical and basolateral membrane voltages, Va and Vbl, respectively, of cells from (1) midguts isolated from the larvae of the oriental armyworm Mythimna separata that were in vitro incubated with toxins and (2) midguts isolated from M. separata larvae force-fed with PSP. We compared the effects of PSP with the effects of the Bacillus thuringiensis toxin Cry1Ab and inactive periplocoside E (PSE) on the midgut epithelial cells. The results showed that Va rapidly decreased in the presence of PSP in a time- and dose-dependent manner, similar to the effects of Cry1Ab. By contrast, PSE did not affect the Va and Vbl. Additionally, PSP did not influence the Vbl. Given these results, we speculate that PSP may modulate transport mechanisms at the apical membrane of the midgut epithelial cells by inhibiting the V-type H+ ATPase.

The purpose of the present study was to observe the response and changes of cranial suture to the distraction forces in growing goats and to examine the expression patterns of TGF-β and BMP during suture distraction.Twenty growing goats were divided into three groups: control (n=4), experimental (n=12), and sham (n=4). A pure titanium distractor was placed in the coronal suture in both the sham and experimental groups. After healing, the distractor was activated for distraction of the coronal suture at a rate of 0.5 mm/day for 8 days in the experimental group. Three animals were killed respectively, at 0，2，4 and 8 weeks after completion of suture distraction. No force was applied in the sham group. X- Ray examination was taken and the coronal suture samples were harvested and processed for histological analysis and scanning electron microscopic analysis and immunohistochemistry of TGF-β and BMP. The coronal sutures of experimental group were separated successfully. Signs of intramembranous bone formation and remodeling were found in the distracted suture，and the sutural structure almost return to its normal state at 8 weeks after end of distraction. At 0 and 2 weeks after completion of suture distraction, the collagen fiber bundles were strengthened and aligned in the direction of the distracted forces. Strong expression of BMP and TGF-β were detected in the fibroblast-like cells and the active osteoblasts. At 4 weeks after suture distraction, signs of intramembranous ossification were found in the edge areas of the distracted suture, and the positive staining of BMP and TGF-β was still noted in the osteoblasts around the newly formed bone trabeculae. This study suggests that cranial suture expansion can be achieved in growing animal by distraction osteogenesis. Mechanical strain resulted from distractor can induce the adaptive remodeling in the cranial suture of growing goats. It also suggests BMP and TGF-β may play very important roles in the process of bone formation and remodeling during suture distraction osteogenesis.

Cranial sutures produce new bone at the sutural edges of the bone fronts in response to external stimuli. Little is known regarding the mechanism of osteogenesis in cranial sutures. Ets1 and Cbfa1 are two important osteogenic transcription factors regulating the differentiation and maturation of osteoblasts. But their function in cranial sutures is not still elucidated. We have investigated the gene expression of Ets1 and Cbfa1 in rat’s calvarial sutural osteoblast-like cells under a single period of mechanical strain. The cells were isolated from the cranial suture of SD rats and cultured in vitro, and subjected to a single 40 minutes mechanical strain using a four-point bending apparatus. The gene expression patterns of Ets1 and Cbfa1 were examined by RT-PCR. Both mRNA levels of Ets1 and Cbfa1 have increased significantly within 6 and 12 hours respectively after mechanical strain were applied, and the increase returned to control level thereafter. However, Ets1 and Cbfa1 exhibited different temporal expression patterns: Ets1 expressed immediately after the mechanical loading and reached the maximum transcription at 0.5h; whereas Cbfa1 experienced a latency period first, then increased slowly within 2 hours, and reached the maximum transcription at 6 h. The maximum transcription of Cbfa1 was about 2.58 fold of that of Ets1. Ets1and Cbfa1 may play different roles in regulating bone matrix protein expressions in osteoblast-like cells during suture distraction and their function is time-dependent. High frequency distraction (>2times/24h) is favourable to the maximal expression of the two genes.

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