1.The effects of rehabilitation training on microvessel nltrastructure and expression of CD31 in rats with focal cerebral infarction
Xiquan HU ; Haiqing ZHENG ; Tiebin YAN ; Sanqiang PAN ; Shangfeng MA
Chinese Journal of Physical Medicine and Rehabilitation 2009;31(2):73-77
Objective To study the effects of rehabilitation training on angiogenesis and its ultrastructure and expression of CD32 in the peri-infarction region of rats with focal cerebral infarction. Methods Sixty-six Spra-gue-Dawley rats with experimental left middle cerebral artery occlusion (MCAO) were used as subjects in this study. All the rats were randomly divided into three groups: a rehabilitation training group (n=30), which was given bar rotating, balancing and rolling exercises everyday after 48 hours post MCAO; a control group (n = 30) and a sham-operated group (n=6), which were fed in cages with no special training exercises. Then brain tissues were fixed on the 3rd d, 7th d, 14th d after MCAO, for observing the morphological alterations of microvessels in peri-infarction re-gion using transmission electron microscope, immunohistochemistry and Western blotting were used to measure the ex-pression of CD31, which acted as the marker of the neogenetic microvessels. Results (1) It showed that the capil-lary endothelial cells were less edematous in the rehabilitation training group, and there were less pinocytosis bullae in basal membrane more integral nucleus of endothelial cells in rehabilitation training group when compared with those in the control group. (2) Expression of CD31 can be observed in peri-infarction region in both groups from the 3rd d on-wards, and peaked on the 7th d, and then gradually went down after the 14th d. Comparison between the 2 groups showed that the expression of CD31 in rehabilitation training group was higher than that in the control group at every time point, but statistical difference between the 2 groups in this regard could be revealed only on the 7th d (P< 0.05), Conclusion Rehabilitation training could promote ultrastruetural recovery of microvessels and induce an-giogenesis in peri-infaretion region, and it might be one of the mechanisms of neural functional recovery in rats after MCAO.
2.The effects of rehabilitative training on motor function and expression of GAP-43 and SYN in rats with local cerebral infarction
Xiquan HU ; Haiqing ZHENG ; Tiebin YAN ; Sanqiang PAN ; Shangfeng MA
Chinese Journal of Physical Medicine and Rehabilitation 2008;30(4):217-221
Objective To study the effects of rehabilitative training on motor function and expression of GAP-43 and SYN in rats with local cerebral infarction. Methods A total of 76 male adult Sprague-Dawley rats were randomly divided into a rehabilitative training group(n=32),a control group(n=32),and a sham-operated group(n=1 2).All the rats were subjected to left middle cerebral artery occlusion(MCAO)with the suture occlu sion.Motor training programs including balancing,grasping,rotating and walking exercises were administered to the rats of the rehabilitative training group at 48 hours post-operation,while those of other two groups were reared in their original living status without any special training.The animals were given behavioral tests with Bederson test,balancing wood test,net screen test to assess the functional outcome,and immunohistochemistry staining was employed to evaluate the exDression of GAP-43 and SYN in peri-infarction cortex at the 3rd,7th,21 st,35th days after MACO,respectively. Results The scores of behavioral tests in the rehabilitative training group was better than those in the control group(P<0.05)at the 7th,21 st,35th day after MCAO,and the immunostaining showed that expression of GAP-43 was higher in the rehabilitative training group than that in the control group(P<0.05)and the sham operated group(P<0.01)at the 7th and 21 st days post-operation,respectively,and that the expression of SYN was higher in the rehabilitative training group than that in the control(P<0.05)and the sham operated groups(P<0.05)at the 21 st and the 35th days post-operation,respectively. Conclusion Rehabilitative training can improve func tional recovery in rats with local cerebral infarction,and the function enhancement may be partially attributed to the up-regulation of expression of GAP-43 and SYN in peri-infarction cortex.
3.Animal experiments of immunotherapy with ovarian tumor vaccine during lymphopenia induced by chemotherapy
Qiling LI ; Yunping WANG ; Shangfeng GAO ; Jan MA ; Caixia FENG ; Yueling WANG
Chinese Journal of Obstetrics and Gynecology 2009;44(11):856-860
Objective To explore the mechanisms and effects of adoptive immunotherapy with ovarian cancer vaccine modified by GM-CSF gene which was used after immunologic reconstitution during lymphopenia induced by chemotherapy.Methods Lymphopenia was induced by chemotherapy with cyclophosphamide.The immune reconstituted model was built in rats.The tumor vaccine draining lymph nodes were harvested after the ovarian cancer cells NUTU-19 modified by GM-CSF gene were injected.The effector T cells (T_E) were got after being stimulated and amplified.Enzyme-linked immunosorbent assay was used to detect the level of interleukin (IL)-2 and IL-4 secreted by T_E.Intracellular cytokine staining was used to determine frequency of tumor-specific T_E.Fluorescence-activated cell sorting (FACS) was used to detect the special cytotoxicity ofT_E killing target cells.The survival period of rats bearing pre-established abdominal ovariam carcinoma after being adoptively transferred byT_E.was observed.Results Compared with those in control group,the significant higher levels IL-2[(65.7±4.0) pg/ml]and lower levels IL-4 [(277±49) pg/ml]were observed in chemotherapy-immune recunstitution-vaccine immunization group.The amount of CD_4~+ T cells secreting interferon-γ (13.0±2.1)% were also significantly increased.The rate of the special cytotoxicity of killing T cells (86.5±1.1) % was markedly improved.The survival period of rats (110±16) days was increased in chemotherapy-immune reconstitution-vaccine immunization group.Conclusions The combined immunotherapy of chemotherapy-immune reconstitution-tumor vaccine immunotherapy may increase the frequency and function of specific tumor T_E.The specific cytotoxicity is increased and the weak reaction of T_E to tumor is improved,which showed that this therapy can enhance immune reaction.