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Objective:To evaluate the effect of sufentanil on activation of Schwann cells after peripheral nerve injury in mice.Methods:Eighty healthy pathogen-free male Balb/c mice, aged 6-8 weeks, weighing 18-22 g, were divided into 4 groups ( n=20 each) using a random number table method: peripheral nerve injury group (group PNI), high dose sufentanil group (group H), medium dose sufentanil group (group M) and low dose sufentanil group (group L). The model of unilateral sciatic nerve transaction was established in ketamine-anesthetized mice.Immediately after establishment of the model, sufentanil 10, 5 and 2.5 μg/kg was injected intraperitoneally once a day for 3 consecutive days in H, M and L groups, respectively, while the equal volume of normal saline was given instead in group PNI.Sciatic function index (SFI) was calculated at 4, 8 and 12 weeks after establishment of the model.At 2, 4, 8 and 12 weeks, 5 mice in each group were sacrificed, and segments of the injuried ipsilateral sciatic nerve were removed for examination of the ultrastructure of the sciatic nerve (with a transmission electron microscope) and for detection of the expression of glial fibrillary acidic protein (GFAP) of sciatic nerve (by immunohistochemistry). Results:Compared with group PNI, SFI was significantly increased, and the expression of GFAP was up-regluated at each time point after establishment of the model in H and M groups ( P<0.05) and no significant change was found in SFI and GFAP expression after establishment of the model in group L ( P>0.05). Compared with group L, SFI was significantly increased, and GFAP expression was up-regluated in H and M groups ( P<0.05). There was no significant difference in SFI and GFAP expression between group H and group M ( P>0.05). The thickness of myelin lamellae was dense, and the proliferation of Schwann cells was not marked in H and M groups.The thickness of myelin lamellae was thin, and the proliferation of Schwann cells was marked in L and MO groups. Conclusion:The mechanism by which sufentanil improves repair after peripheral nerve injury may be related to promoting activation of Schwann cells in mice.
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Objective To evaluate the effect of sufentanil on apoptosis in spinal cord neurons of mice with peripheral nerve injury. Methods One hundred and fifty clean-grade healthy male BALB∕c mice, aged 6-8 weeks, weighing 18-22 g, were divided into 3 groups ( n=50 each) using a random number table method: sham operation group (group Sham), peripheral nerve injury group (group PNI) and sufentanil group ( group SF) . The model of unilateral sciatic nerve injury was established in PNI and SF groups. After establishing the model, sufentanil 5. 0 μg∕kg was intraperitoneally injected once a day for 3 consecutive days in group SF, while the equal volume of normal saline was given instead of sufentanil in Sham and PNI groups. Five mice in each group were sacrificed at days 1, 3, 7, 14 and 28 after surgery ( T0-4 ) , and L4-6 segments of the injure ipsilateral spinal cord were removed for examination of pathological changes ( with a light microscope) and for determination of neuronal apoptosis ( by TUNEL assay) . The ap-optosis index ( AI) was calculated. Five mice in each group were sacrificed at T0-4 , and L4-6 segments of the injured ipsilateral spinal cord were removed for detection of the expression of Bcl-2, Bax and cleaved caspase-3 by Western blot. The ratio of Bcl-2 expression to Bax expression ( Bcl-2∕Bax ratio) was calculat-ed. Results Compared with group Sham, the AI was significantly increased, the expression of Bcl-2 pro-tein was down-regulated, and the expression of cleaved caspase-3 and Bax was up-regulated in PNI and SF groups ( P<0. 05) . Compared with group PNI, the AI was significantly decreased, the expression of Bcl-2 protein was up-regulated, the expression of cleaved caspase-3 and Bax was down-regulated, the Bcl-2∕Bax ratio was increased (P<0. 05), and the pathological changes were significantly attenuated in group SF. Conclusion Sufentanil can inhibit apoptosis in spinal cord neurons of mice with peripheral nerve injury.
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OBJECTIVE:To establish UPLC fingerprint of Glycyrrhiza uralensis. METHODS:UPLC method was adopted. The determination was performed on Waters CORTECS UPLC C18column with mobile phase consisted of acetonitrile-0.1% formic acid(gradient elution)at the flow rate of 0.3 mL/min. The detection wavelength was set at 254 nm,and column temperature was 30 ℃. The sample size was 1 μ L. Using glycyrrhizic acid as control,UPLC chromatograms of 27 batches of sample were determined. Similarity evaluation was conducted by using TCM Chromatogram Fingerprint Similarity Evaluation System(2004 A edition)to determine common peak and conduct cluster analysis of 27 batches of samples. RESULTS:There were 20 common peaks in UPLC chromatograms of 27 batches of samples. The similarity degree of S2,S4,S19,S21,S22,S24 were less than 0.90, the others samples were more than 0.90.After validation,UPLC chromatograms of 21 batches of batches of samples were in good agreement with control fingerprint. 27 batches of samples were clustered into 3 categories,in which S24 was categoryⅠ;S2,S4, S12,S19,S21,S22 were categoryⅡ;other were categoryⅢ. CONCLUSIONS:Established fingerprint can provide reference for quality evaluation of G.uralensis.
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Objective To investigate the effects of sufentanil on activation of spinal astrocytes in mice after unilateral sciatic nerve injury.Methods Eighty healthy male BALB/c mice,aged 6-8 weeks,weighing 18-22 g,were randomly divided into 4 groups (n=20 each): sufentanil high dose group (group H),middle dose group (group M),low dose group (group L) and model group (group MO).After model of sciatic nerve injury was established by unilateral sciatic nerve transection,group H,M and L,sufentanil 10 μg/kg,5 μg/kg,2.5 μg/kg was injected intraperitoneally once a day continuously for 3 days,while the equal volume of saline was injected in group MO.Five mice in each group were selected,and the sciatic nerve functional index were measured at 4,8,12 weeks.The 5 mice were sacrificed from each group and the damage on the same side L4-L6 segments of the spinal cord were removed at 2,4,8,12 weeks.The pathological changes were examined under light microscope at 8 week point.The expression of GFAP was determinatied at each time points by immuno-histochemistry.Results Compared with group L and MO,sciatic nerve functional index significantly was increased in groups H and M (P<0.05),and no significantly change was found in group L.Spinal cord neurons had a better morphology in groups H and M than in group L and MO.Compared with group MO,The expression of GFAP were significantly up-regulated in groups H,M and L (P<0.05).Compared with group L,the expression of GFAP were significantly up-regulated in groups H and M (P<0.05).Conclusion Sufentanil promotes spinal astrocyte activation after peripheral nerve injury in mice and improves repair after peripheral nerve injury.
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Objective To evaluate the effect of sufentanil on the expression of basic fibroblast growth factor(bFGF)in nerve tissues after peripheral nerve injury in mice. Methods One hundred pathogen-free healthy male Balb∕c mice, aged 6-8 weeks, weighing 18-22 g, were divided into 4 groups (n=25 each)using a random number table: peripherial nerve injury group(group PNI), high dose sufentanil group(group H), medium dose sufentanil group(group M)and low dose sufentanil group (group L). The model of unilateral sciatic nerve transaction was established in ketamine-anesthetized mice. Sufentanil 100, 50 and 25 μg∕kg were intraperitoneally injected immediately after establishment of the model once a day for 3 consecutive days in H, M and L groups, respectively. The equal volume of normal saline was given instead in group PNI. On 1, 2, 4, 8 and 12 weeks after establishment of the model, 5 mice were sacrificed and the nerve tissues were obtained from the site 05 cm up and down the lesion site of the nerve for examination of the shape of the myelin sheath of nerve fibers(with an electronic microscope).The expression of bFGF in the sciatic nerve tissue was detected by Western blot. Results The shape of medulla sheath was irregular, the thickness of myelin lamellae was thin, the separation of myelin lamellae was aggravate, demyelinate was found, and the proliferation of Schwann cells was not marked in group PNI. The shape of medulla sheath was regular, the thickness of myelin lamellae was dense, and the proliferation of Schwann cells was marked in group H. The shape of medulla sheath was irregular, the separation of mye-lin lamellae was observed, demyelinate was found, and the proliferation of Schwann cells was not marked in group L. Compared with group PNI, the expression of bFGF was significantly up-regulated in H, M and L groups(P<005). Compared with group L, the expression of bFGF was significantly up-regulated in H and M groups(P<005). Compared with group M, the expression of bFGF was significantly up-regulated in group H(P<005). Conclusion The mechanism by which sufentanil improves regeneration and repair after peripheral nerve injury may be related to up-regulating the expression of bFGF in nerve tissues of mice.
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Objective To evaluate the effects of sufentanil on regeneration and repair after single sciatic nerve injury in mice. Methods Seventy?five healthy male BALB∕c mice, aged 6-8 weeks, weig?hing 18-22 g, were divided into 5 groups ( n=15 each) using a random number table: peripherial nerve injury group (group PNI), low, medium and high doses of sufentanil groups (L, M and H groups) and cyclosporine A group ( group C) . The model of unilateral sciatic nerve transection was established in the 5 groups. In L, M and H groups, sufentanil 2?5, 5?0 and 10?0 μg∕kg were injected intraperitoneally, re?spectively, once a day for 3 consecutive days. Cyclosporine A 50 mg∕kg was injected intraperitoneally once a day for 3 consecutive days in group C. The equal volume of normal saline was given once a day for 3 con?secutive days in group PNI. Neurophysiological monitoring was performed at 4, 8 and 12 weeks after opera?tion, the amplitude of compound muscle action potentials of the sciatic nerve was recorded, and the nerve conduction velocity was measured. At 8 weeks after operation, 5 mice were sacrificed, the sciatic nerve 0?5 cm of the upper and lower the anastomosed site was removed for examination of the morphology of myelin sheath with light microscope, and the number of nerve fibers was calculated. Results Compared with group PNI, the amplitude of compound muscle action potentials of the sciatic nerve, nerve conduction ve?locity and the number of nerve fibers was were significantly increased in M, H and C groups ( P0?05 ) . Myelin sheath arrangement was severely irregular, and more vacuoles were found in group PNI. Myelin sheath ar?rangement was irregular, and more vacuoles were found in group L. Myelin sheath arrangement was mainly regular, and vacuoles were found occasionally in group M. Myelin sheath arrangement was regular, and no vacuoles were observed in H and C groups. Conclusion Sufentanil can promote regeneration and repair af?ter peripheral nerve injury in mice.
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Equipping tumor-targeting carrier with cell-penetrating peptide with high transduction efficacy has become a trend. According to the fusion modes and tumor-targeting mechanisms, cell-penetrating peptides can be divided into five categories: first, self-targeting cell-penetrating peptides; second, fusion carriers made of targeting ligands and cell-penetrating peptides; third, cell-penetrating peptide-modified nano-carrier; fourth, tumor-microenvironment-targeting cell-penetrating peptides; fifth, other special cell-penetrating peptides. Fusion carriers, which can not be effectively released into the cytoplasm after transducted into target cells, can be further modified to increase their efficacy. In all, cell-penetrating peptide introduced into tumor-targeting carrier should provide a new era for anti-tumor pharmacy research and cancer therapy.