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Objective@#To investigate the effect of argatroban in repair of spinal cord injury in rats.@*Methods@#A total of 54 female Wistar rats were selected and divided into three groups according to the random number table: sham group, injury group and Argatroban group, with 18 rats in each group. The sham group only took the T10lamina; the injury group used the spinal cord injury device to make the rat spinal cord injury model; the Argatroban group received Argatroban treatment after spinal cord injury. The recovery of hindlimb motor function was evaluated by BBB score and clined plate test before injury and 7, 14, 21, 28, 35 and 42 days after injury. The sensory evoked potentials (SEP) and motor evoked potentials (MEP) were detected 42 days after operation. HE staining was used to compare the size of the cavity in the local region 42 days after injury.@*Results@#At day 7 after injury, the BBB score was (3.7±0.5)points and the inclined plane test was (28.0±2.6)° in the Argatroban group, which were better than those in the injury group [(3.3±0.5)points, (24.3±1.9)°] (P<0.05). At day 42 after injury, the BBB score was (13.0±0.8)points and inclined plane test was (50.7±2.7)° in the Argatroban group, which were significantly better than those in the injury group [(9.7±1.3) points, (40.5±2.7)°] (P<0.05). But all the above values in the Argatroban group were significantly lower than those in the sham group [(21.0±0.0)points, (60.0±0.0)°](P<0.05). At day 42 after operation, the SEP latency [(25.0±0.9)ms] in the Argatroban group was significantly shorter than that in the injury group [(31.5±1.9) ms]; the amplitude [(2.1±0.1)μV] in the Argatroban group was lower than that in the injury group [(0.5±0.1)μV] (P<0.05). The MEP latency [(11.5±1.0)ms] in the Argatroban group was significantly shorter than that in the injury group [(17.5±1.1)ms], and the amplitude [(4.8±0.8)μV] in the Argatroban group was lower than that in the injury group [(2.8±0.7)μV] (P<0.05). And the SEP or MEP latency and amplitude in the Argatroban group showed significant differences compared to the sham group [(7.5±1.0)ms, (7.5±1.0)μV](P<0.05). HE staining showed that the central area of the lesion in the Argatroban group [(0.35±0.04)mm2] was significantly smaller than that in the injury group [(0.71±0.05)mm2].@*Conclusion@#After spinal cord injury, argatroban can protect the spinal cord tissue effectively in the injured area and promote recovery of sensory and motor function in the hind limbs of rats.
ABSTRACT
Objective To investigate the effect of argatroban in repair of spinal cord injury in rats.Methods A total of 54 female Wistar rats were selected and divided into three groups according to the random number table:sham group,injury group and Argatroban group,with 18 rats in each group.The sham group only took the T101amina;the injury group used the spinal cord injury device to make the rat spinal cord injury model;the Argatroban group received Argatroban treatment after spinal cord injury.The recovery of hindlimb motor function was evaluated by BBB score and clined plate test before injury and 7,14,21,28,35 and 42 days after injury.The sensory evoked potentials (SEP) and motor evoked potentials (MEP) were detected 42 days after operation.HE staining was used to compare the size of the cavity in the local region 42 days after injury.Results At day 7 after injury,the BBB score was (3.7 ±0.5) points and the inclined plane test was (28.0 ± 2.6) ° in the Argatroban group,which were better than those in the injury group [(3.3 ± 0.5) points,(24.3 ± 1.9) °] (p ＜ 0.05).At day 42 after injury,the BBB score was (13.0 ± 0.8) points and inclined plane test was (50.7 ± 2.7) ° in the Argatroban group,which were significantly better than those in the injury group [(9.7 ± 1.3) points,(40.5 ± 2.7)°] (p ＜0.05).But all the above values in the Argatroban group were significantly lower than those in the sham group [(21.0 ± 0.0) points,(60.0 ± 0.0) °] (P ＜ 0.05).At day 42 after operation,the SEP latency [(25.0 ± 0.9)ms] in the Argatroban group was significantly shorter than that in the injury group [(31.5 ± 1.9) ms];the amplitude [(2.1 ± 0.1) μV] in the Argatroban group was lower than that in the injury group [(0.5 ± 0.1) μV] (P ＜ 0.05).The MEP latency [(11.5 ± 1.0) ms]in the Argatroban group was significantly shorter than that in the injury group [(17.5 ± 1.1) ms],and the amplitude [(4.8 ± 0.8) μV] in the Argatroban group was lower than that in the injury group [(2.8 ± 0.7) μV] (P ＜ 0.05).And the SEP or MEP latency and amplitude in the Argatroban group showed significant differences compared to the sham group [(7.5 ± 1.0) ms,(7.5 ± 1.0) μV] (P ＜0.05) . HE staining showed that the central area of the lesion in the Argatroban group [(0.35 ± 0.04) mm2]was significantly smaller than that in the injury group [(0.71 ± 0.05)mm2].Conclusion After spinal cord injury,argatroban can protect the spinal cord tissue effectively in the injured area and promote recovery of sensory and motor function in the hind limbs of rats.
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Objective To design a new cancer vaccine by using alpha-galactosylceramide (α-Galcer,α-GC) loaded tumor cells in combination with TLR 9 ligand and to evaluate its therapeutic effects on colon canc-er in mice.Methods MC38 cells were transfected with lentivirus (GFP-CD1d) to prepare CD1d-MC38 cells. The expression of CD1d molecules in CD1d-MC38 cells was detected by fluorescence microscopy , RT-PCR and flow cytometry.The sorted CD1d-MC38 cells were loaded with α-Galcer to prepare CD1d-MC38/α-GC complex. Flow cytometry was performed to evaluate the efficiency of combination .A mouse model of colon cancer was es-tablished to investigate the therapeutic effects of α-Galcer loaded tumor cells in combination with TLR 9 ligand ( CD1d-MC38/α-GC+CpG1826) on colon cancer in mice by analyzing tumor growth and mice survival time .Im-munohistochemical staining was used to detect CD 4+T and CD8+T infiltrating lymphocytes in tumor tissues .Re-sults The MC38 cancer cells that expressed CD 1d and GFP were successfully constructed , among which 98.10%±2.53%were positive for CD1d.Moreover, the CD1d-MC38 cells could combine with α-Galcer effec-tively in a dose and time dependent manner .Compared with PBS treated group ,α-GC treated group and TLR9 ligand treated group , the experimental vaccine strategy was sufficient to inhibit the growth of established tumors and prolong survival of tumor-bearing mice (P<0.01).Immunohistochemistry analysis revealed that levels of CD4+T cells and CD8+T cells in experiment group were significantly higher than those in groups treated with PBS,α-GC and TLR9 ligand (P<0.01).Conclusion CD1d-MC38/α-GC in combination with CpG1826 could efficiently inhibit the growth of established tumors and prolong survival of tumor-bearing mice .Immunohisto-chemistry analysis revealed that CD 4+T cells and CD8+T cells played important roles in anti-tumor immunity.
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In vivo,dendritic cells(DCs)are the most powerful antigen-presenting cells (APCs).DCs based neoplasm immunotherapy has recently become the focus of research.DCs can be used not only as separate cell vaccine to mediate anti-tumor immunity,but can also be combined with cytokines,immune cells,and other methods in tumor immunotherapy. In animal models and clinical trials,DC-based immunotherapy as well as other tumor immunotherapy strategies have gained more significant progress.
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Objective:To prepare functional monoclonal antibodies against human 4-1BBL molecule and analysis of their biological characteristics.Methods:Female BALB/c mice of 6-8 weeks old were immunized with 4-1BBL transfectant (L929/4-1BBL) as immunogen.The spleen B cells of the mice were fused with sp2/0 and hybridoma cells were screened with 4-1BBL transfectant (L929/4-1BBL) by FCM.The biological characteristics of antibody were investigated by rapid isotyping analysis,karyotype analysis,competitive inhibition test etc.Furthermore,the growth of monocytes in vitro was determined by cell number counting and the cytokine concentration in the supernatants was assayed by ELISA.Results:One hybridoma cell line named 3E7 was obtained,which had the property of secreting anti-human 4-1BBL monoclonal antibody continuously and steadily.This mAb specifically recognized human 4-1BBL molecules.The experimental results manifested that mAb 3E7 could effectively enhance the growth of monocytes and the high level IL-6 and TNF-? secretion.Conclusion:One hybridoma cell line which can secret a functional mouse anti-human 4-1BBL mAb has been developed successfully.This mAb can specifically recognize human 4-1BBL and regulate growth and functions of monocytes in vitro.