Objective To investigate the influence of different transplantating times on the survival and immigration of the bone marrow mesenchymal stem cells (BMSCs) in injured spinal cord by subarachnoid administration, and to evaluate the most optimal subarachnoid administration times for BMSCs. Methods Eight adult male rats (weighing 120 g) were used to isolate BMSCs that were cultured, purified and labeled with Hoechst 33342 in vitro. Another 75 adult Wistar rats (weighing 220 g) were made the spinal cord injury (SCI) models at T9,10 level according to the improved Allen’s method and were randomly divided into 5 groups (groups A, B, C, D, and E, n=15). The labeled BMSCs at 1 × 107/mL 0.1 mL were injected into subarachnoid space of the rats via a catheters under the subarachnoid space in groups A (one time at 1 week), B ( two times at 1 and 3 weeks), C (3 times at 1, 3, and 5 weeks) and D (5 times at 1, 3, 5, 7, and 9 weeks) and 0.2 mL phosphate-buffered sal ine (PBS) was injected in group E (5 times at 1, 3, 5, 7, and 9 weeks) as blank control. The neurological functions were evaluated using the Basso-Beattie-Bresnahan (BBB) scale 1, 3, 5, 7, 9, and 12 weeks after transplantation. The migration, survival, differentiation, and histomorphological changes of BMSCs were observed by HE, immunohistochemistry, and fluorescence microscopy. Results At 3 weeks after injury, there were significant differences in the BBB scores between group E and groups A, B, C, D (P lt; 0.01), and between groups A, B and groups C, D (P lt; 0.01). At 7, 9, and 12 weeks, the BBB scores were significantly higher in groups C and D than in groups A and B (P lt; 0.01), and in group B than in group A (P lt; 0.01). There were no significant differences in the BBB scores between groups C and D (P gt; 0.05). The fluorescence microscopy showed that the transplanted BMSCs survived and grew in the injured region at 3 weeks after injury and as time went on, the transplanted cells gradually decreased in group A; in groups B, C, and D, BMSCs count reached the peak values at 5 and 7 weeks and then gradually decreased. At 12 weeks, the survival BMSCs were significantly more in groups C and D than in groups A and B (P lt; 0.01). HE staining showed that the formation of cavity was observed in each group at 3 weeks after injury and the area of cavity gradually decreased in groups A, B, C, and D. At 12 weeks, the area of cavity was the miximal in groups C and D, moderate in groups A and B, and the maximal in group E. The immunohistochemistry staining indicated that the expression of NF-200 was more intense in groups C and D than in groups A and B. The expression of NF-200-positive fibers was more intense in group C. Conclusion Multiple administration of BMSCs promotes the restoration of injured spinal cord and improves neurological functions, and three times for BMSCs transplantation is best
Objective To investigate the therapeutic effects of transplanting allogeneic marrow mesenchymal stem cells (MSCs) via subarachnoid space on spinal cord injury(SCI) and the T cell subpopulation. Methods Density gradient centrifugation was used to isolate and expand MSCs from bone marrow of 10 six-week-old SD rats. The SCI model was produced by weightbeating from 60 eight-week-old female SD rats. Forty survival SCI rats,which BBB scores were zero, were divided randomly into 2 groups:experimental group(group A) and control group(group B). In addition, 20 normal eightweekold SD ratswere used as blank group (group C). In group A, 1 ml cells suspention containing MSCs(the 6th generation, 2×106/ml) was injected via subarachnoid space. Ingroup B, equal volume of L-DMEM was injected in the same way. The BBB score was obtained after 1st,2nd and 3rd weeks of injection. At the same time,T cell subpopulation was detected by flow cytometry. Results The BBB score in group A was better than that in group B, but fewer than that in group C in the 3rd week. CD4+T cells in group A were less than those in groups B and C in the 1st, 2nd, and 3rd weeks. CD8+T cells in group A were less than those in groups B and C in the 2nd and 3rd weeks. The ratio of CD4+/CD8+T cells in group A was less than those in groups B and C in the 1st week. Above differences showed statistically significant difference(P＜0.05). However, there were no statistically significant differences in the ratio of CD4+/CD8+T cells between group A and groups B, C in the 2nd and 3rd weeks (P＞0.05). Conclusion The above results suggest that allogeneic MSCs transplantation via subarachnoid space is beneficial to SCI to some extend, do not result in rejection in vivo. Furthermore, it can lead to immunosuppression in short time. So, it provides clues to apply MSCs to treat SCI and other diseases.
目的:探讨住院期间自发性蛛网膜下腔出血患者再出血的相关因素和对策。方法:对42例自发性蛛网膜下腔出血患者住院期间引起再出血的各种相关因素进行分析。结果: 再出血危险因素:34例持续高血压,14例情绪激动,7例下床排便用力,6例剧烈活动,7例睡眠不佳烦燥后出血,4例再出血发生在术前8小时, 4例患者在腰穿时出血。结论: 自发性蛛网膜下腔出血患者住院时高血压和和各种引起血压升高的诱因是引起再出血最主要原因．同时出血时间间隔和住院期间不适当的腰穿也是引起再出血的重要原因。
Objective To explore the effectiveness and safety of exchange of cerebrospinal fluid in the treatment of subarachnoid hemorrhage （SAH）. Methods Sixty SAH patients diagnosed by CT and lumbarpuncture were randomly assigned into a control group （n =30, received conventional treatment） and a treatment group （n =30, received exchange of cerebrospinal fluid plus conventional treatment）. The main complications and effectiveness between the two groups were compared. SPSS 10.0 was used for statistical analysis. Results Compared with the control group, complications of persistent headache （ P =0.002 and 0. 007 respectively）, cerebral vasospasm （ P =0. 028 ） and hydrocephalus （ P =0. 038 ） were fewer in the treatment group. No significant difference in the incidence of rehaemorrhagia was found between the two groups （P = 1. 000）. Better effectiveness was observed in the treatment group （RR. 3.00, 95% CI 1. 014 to 8. 880, P = 0. 044 ）. Conclusions Exchange of cerebrospinal fluid plus conventional treatment is more effective than conventional treatment alone in the treatment of SAH.
ObjectiveTo assess the effectiveness and safety of continuous lumbar drainage of cerebrospinal fluid for subarachnoid hemorrhage (SAH). MethodsThe Cochrane Library (January 1992 to May 2013), Medline (January 1950 to May 2013), SinoMed (January 1979 to May 2013), CNKI (January 1979 to May 2013), and Wanfang Database (January 1979 to May 2013) were searched for randomized controlled trials (RCTs) on continuous lumbar drainage for SAH. The method of Cochrane systematic review was used to evaluate all the included RCTs. ResultsTwelve RCTs (857 patients) met the inclusion criteria, but the general methodological quality of trials was poor. Only two studies addressed the outcomes about SAH patients' death, vegetative state or disability and other adverse events at the end of the follow-up period (at least 3 months). Meta-analysis of fatality showed a better effect of lumbar continuous drainage of the cerebrospinal fluid for SAH than the control group with statistical significance [RR=0.27, 95%CI (0.12, 0.59), P=0.001]. Meta-analysis of complications also showed a better effect of lumbar continuous drainage of the cerebrospinal fluid for SAH than the control group with statistical significance (except re-bleeding) [cerebral vasospasm: RR=0.20, 95%CI (0.14, 0.30), P<0.000 01; hydrocephalus: RR=0.24, 95%CI (0.13, 0.41), P<0.000 01; cerebral infarction: RR=0.27, 95%CI (0.16, 0.45), P<0.000 01]. Only one trial reported the adverse events of lumbar continuous drainage of the cerebrospinal fluid including intracranial infection and intracranial hypotension reaction, while the others did not report the adverse events. ConclusionWith poor quality of the most included trials, insufficient evidence is obtained to support the conclusion that lumbar continuous drainage of the cerebrospinal fluid is safe and effective in the treatment of SAH. Further high-quality RCTs should be carried out to provide more reliable evidences.