Jmjd3和Ezh2拮抗调节小鼠骨折愈合

目的:探讨Jmjd3和Ezh2在小鼠骨折愈合过程中的作用。方法:以软骨细胞条件性基因敲除8-10周龄小鼠为研究对象,按基因型随机分为6组,每组5只:其中实验组基因型为Jmjd3~(fl/fl)/Col2a1-Cre ~(ERT2),Ezh2~(fl/fl)/Col2a1-Cre ~(ERT2)或Jmjd~(3fl/fl)/Ezh2~(fl/fl)/Col2a1-Cre ~(ERT2);对照组基因型为Jmjd3~(fl/fl),Ezh2~(fl/fl)或Jmjd3~(fl/fl)/Ezh2~(fl/fl)。建立骨髓腔中插入固定针的稳定性胫骨骨折模型,于骨折术后3天、5天和7天腹腔注射Tamoxifen 3 mg/次/天。各组于术后3W处死,并于骨折部位取材行X线片及组织学检查。结果:通过连续的X线影像学及HE组织切片观察,骨折术后3周是判断小鼠骨折愈合情况的最佳时间点。X线片发现骨折术后3W时软骨细胞内Jmjd3被敲除小鼠的骨折线较对照组明显且骨化骨痂大小和密度均较低,HE切片显示骨化骨痂面积显著低于对照组,而软骨骨痂面积高于对照组;相反,X线片发现Ezh2被敲除小鼠的骨痂面积明显大于对照组,且密度高于对照组,HE组织切片显示Ezh2被敲除的小鼠的骨化骨痂的钙化程度更高,骨小梁更粗更密集。最后,X线片和HE切片均没有发现软骨细胞Jmjd3和Ezh2同时被敲除的小鼠与对照小鼠之间存在明显差异。结论:以软骨细胞特异基因敲除小鼠为基础,我们首次发现Jmjd3具有促进骨折愈合的作用,而Ezh2具有抑制骨折愈合的作用;并且发现Jmjd3和Ezh2对抗调节小鼠的骨折愈合过程,这些发现为骨折愈合治疗提供了新的分子实验基础。     

人脐带间充质干细胞对脊柱骨折大鼠愈合及神经功能的影响

摘要 目的：探讨人脐带间充质干细胞(Human umbilical cord mesenchymal stem cells,hUC-MSCs)对脊柱骨折大鼠愈合及神经功能的影响。方法：脊柱骨折Sprague-Dawley雄性大鼠模型30只随机分为hUC-MSCs组与对照组,各15只。hUC-MSCs组大鼠在骨折部位移植0.5 mL的hUC-MSCs(细胞浓度为2×10⁶/mL),对照组大鼠移植同体积的生理盐水,记录大鼠愈合及神经功能变化情况。结果：两组造模后15 min、30 min、90 min的平均动脉压都波动明显,不过组间对比差异无统计学意义(P>0.05)。与造模后2 w对比,两组造模后4 w的神经功能BBB评分均升高,且hUC-MSCs组造模后2 w、4 w的神经功能BBB评分都高于对照组(P<0.05)。hUC-MSCs组造模后8 w的骨体积分数高于对照组(P<0.05)。hUC-MSCs组骨折部位附近有少量骨痂生长,骨折线逐渐消失;骨痂已明显包裹骨折部位。hUC-MSCs组造模后8 w的脊髓细胞凋亡指数低于对照组(P<0.05)。结论：hUC-MSCs在脊柱骨折大鼠的应用能促进骨折愈合与改善神经功能,也可以抑制脊髓细胞凋亡,从而发挥很好的治疗作用。     

Upregulation of inflammatory genes and downregulation of sclerostin gene expression are key elements in the early phase of fragility fracture healing

Background

Fracture healing is orchestrated by a specific set of events that culminates in the repair of bone and reachievement of its biomechanical properties. The aim of our work was to study the sequence of gene expression events involved in inflammation and bone remodeling occurring in the early phases of callus formation in osteoporotic patients.

Methodology/Principal Findings

Fifty-six patients submitted to hip replacement surgery after a low-energy hip fracture were enrolled in this study. The patients were grouped according to the time interval between fracture and surgery: bone collected within 3 days after fracture (n = 13); between the 4^th and 7^th day (n = 33); and after one week from the fracture (n = 10). Inflammation- and bone metabolism-related genes were assessed at the fracture site. The expression of pro-inflammatory cytokines was increased in the first days after fracture. The genes responsible for bone formation and resorption were upregulated one week after fracture. The increase in RANKL expression occurred just before that, between the 4^th–7^th days after fracture. Sclerostin expression diminished during the first days after fracture.

Conclusions

The expression of inflammation-related genes, especially IL-6, is highest at the very first days after fracture but from day 4 onwards there is a shift towards bone remodeling genes, suggesting that the inflammatory phase triggers bone healing. We propose that an initial inflammatory stimulus and a decrease in sclerostin-related effects are the key components in fracture healing. In osteoporotic patients, cellular machinery seems to adequately react to the inflammatory stimulus, therefore local promotion of these events might constitute a promising medical intervention to accelerate fracture healing.     

髓腔内固定联合低强度脉冲超声对兔股骨中段骨折愈合作用研究

摘要 目的：针对髓腔内固定联合低强度脉冲超声对兔股骨中段骨折愈合作用进行研究。方法：选择成年兔股骨中段骨折40只,作为本次的研究对象,将其平均分为对照组和观察组。所有兔子,在手术前禁水、禁食,对其右后肢进行备皮,称重、麻醉,对照组实施髓腔内固定治疗,观察组实施髓腔内固定联合低强度脉冲超声治疗。治疗后1、2、3、4周对兔子的骨折部位进行影像学检查确定兔子的骨折线模糊情况,并在各周采集样品进行组织学检查。治疗4周后对骨折愈合情况进行检查。结果：观察组愈合程度I级、II级、II级比例均低于对照组相应比例,差异具有统计学意义（P＜0.05）,观察组IV级、V级比例分别为35.0 %、55.0 %,均高于对照组的5.0 %、5.0 %,差异具有统计学意义（P＜0.05）;观察组兔子的在1 w、2 w、3 w、4 w骨折线模糊程度评分高于对照组相应时间评分,差异具有统计学意义（P＜0.05）。结论：在兔股骨中断骨折治疗中,实施髓腔内固定联合低强度脉冲超声,可以提高骨折的愈合速度,加速骨折修复,整体治疗效果显著,可以在临床上进行推广实施。     

Autologous mesenchymal stem cells loaded in Gelfoam<Superscript>®</Superscript> for structural bone allograft healing in rabbits

This study was designed to evaluate the effect of autologous bone marrow mesenchymal stem cells (MSCs) seeded into Gelfoam^® on structural bone allograft healing. Thirty New Zealand white rabbits were divided into two groups. Segmental bone defect was created on diaphysis of the femur, and the defect was reconstructed with structural bone allograft. In experimental group, structural allograft was wrapped around by Gelfoam^® containing autologous MSCs, whereas cells were not included in control group. At 4, 8, 12 weeks, the femur of rabbits underwent radiographic and histologic evaluation for bony union. Bone morphogenic protein-2 (BMP-2), BMP-4, BMP-7, vascular endothelial growth factor (VEGF), and receptor activator of nuclear factor-kappa B ligand (RANKL) were measured within the grafted periosteal tissue. Bony union was not achieved in both groups at 4 and 8 weeks. At 12 weeks, three out of five femurs in experimental group were united, but one out of five in control group was united. Mean Taira scores were significantly different between two groups. The expression of BMP-2 was significantly higher at 4, 8 weeks, the expressions of BMP-4 and BMP-7 were significantly higher at 8 and 12 weeks, and the expression of VEGF and RANKL were significantly higher at all time points in experimental group. Incorporation of the structural bone allograft could be enhanced if allograft is covered with Gelfoam^® containing autologous MSCs. MSCs have influence on not only bone formation, but neo-angiogenesis, and bone resorption.     

Enhancement of tendon-bone healing with the use of bone morphogenetic protein-2 inserted into the suture anchor hole in a rabbit patellar tendon model

Background aimsSuture anchor fixation failure has been reported as a result of anchor loosening and migration during the tendon-bone repair. The aim of this study was to evaluate the effects of bone morphogenetic protein-2 (BMP-2) inserted into the suture anchor hole on bone formation and the tendon-bone healing.MethodsBoth back legs of 24 New Zealand White rabbits (n = 48) were used in this study. A metal suture anchor was then placed 5 mm below the cortex. In the control group, the space over the eyelet of the anchor (suture anchor hole) was not filled. In the experimental group, the suture anchor hole was filled with 0.1 mL of fibrin glue (group 2) or collagen gel (group 3) with 1 μg BMP-2. Histologic analysis, real-time-polymerase chain reaction, bone density and failure load measurement were performed, and differences were analyzed at 4 and 8 weeks.ResultsHistologic analysis revealed more abundant new bone, mature bone and organized fibrocartilage at the tendon-bone interface at 4 and 8 weeks in groups in which BMP-2 was applied. At 8 weeks, the failure load of groups 1, 2 and 3 was significantly different among the three groups (P = 0.01). After post hoc Tukey test, the failure load of group 2 was significantly higher than that of group 1 (P = 0.01).ConclusionsBMP-2, administrated as described in this study, improved tendon-bone healing and bone formation, resulting in improved biomechanical strength of the tendon-bone junction.     

Organic Gallium Treatment Improves Osteoporotic Fracture Healing Through Affecting the OPG/RANKL Ratio and Expression of Serum Inflammatory Cytokines in Ovariectomized Rats

This study aimed to investigate the impact of organic gallium (OG) on osteoporotic fracture healing in ovariectomized female Sprague-Dawley rats, as well as study the mechanisms of OG on osteoporotic fracture healing. Forty-five female Sprague-Dawley rats were divided into three groups: sham operation group (Sxas control group), ovariectomized group (Ovx), and Ovx treated with OG group (Ovx + OG). Rat femoral fractures were studied using a standardized fracture-healing model utilizing bone fixation with an intramedullary pin. Six weeks later, analyses of micro-CT, histomorphometric, RNA extraction, RT-qPCR, and serum were performed following sacrifice of all mice. In comparison with Ovx group, OG can significantly increase bone volume (BV), tissue volume (TV), BV/TV radio, bone strength, callus bony area, and as similar to BMP-2 expression. OG treatment elevated OPG messenger RNA (mRNA) and inhibited RANKL mRNA, and showed an effect on OPG/RANKL ratio. OG treatment can inhibit the expression of TNF-α and IL-6. In conclusion, current study results indicate that organic OG can positively affect the OPG/RANKL ratio and inhibit the expression of serum inflammatory cytokines; thus, it can improve osteoporotic fracture healing.     

Spinal cord injury regeneration using autologous bone marrow-derived neurocytes and rat embryonic stem cells: A comparative study in rats

BACKGROUNDSpinal cord injury (SCI) is an important cause of traumatic paralysis and is mainly due to motor vehicle accidents. However, there is no definite treatment for spinal cord damage.AIMTo assess the outcome of rat embryonic stem cells (rESC) and autologous bone marrow-derived neurocytes (ABMDN) treatment in iatrogenic SCI created in rats, and to compare the efficacy of the two different cell types.METHODSThe study comprised 45 male Wistar rats weighing between 250 and 300 g, which were divided into three groups, the control, rESC and ABMDN groups. The anesthetized animals underwent exposure of the thoracic 8^th to lumbar 1^st vertebrae. A T10-thoracic 12^th vertebrae laminectomy was performed to expose the spinal cord. A drop-weight injury using a 10 g weight from a height of 25 cm onto the exposed spinal cord was conducted. The wound was closed in layers. The urinary bladder was manually evacuated twice daily and after each evacuation Ringer lactate 5 mL/100 g was administered, twice daily after each bladder evacuation for the first 7 postoperative days. On the 10^th day, the rats underwent nerve conduction studies and behavioral assessment [Basso, Beattie, Brenham (BBB)] to confirm paraplegia. Rat embryonic stem cells, ABMDN and saline were injected on the 10^th day. The animals were euthanized after 8 wk and the spinal cord was isolated, removed and placed in 2% formalin for histopathological analysis to assess the healing of neural tissues at the axonal level.RESULTSAll the animals tolerated the procedure well. The BBB scale scoring showed that at the end of the first week no recovery was observed in the groups. Post-injection, there was a strong and significant improvement in rats receiving rESC and ABMDN as compared to the control group based on the BBB scale, and the Train-of-four-Watch SX acceleromyography device exhibited statistically significant (P < 0.0001) regeneration of neural tissue after SCI. Histological evaluation of the spinal cord showed maximum vacuolization and least gliosis in the control group compared to the rESC and ABMDN treated animals. In the ABMDN group, limited vacuolization and more prominent gliosis were observed in all specimens as compared to the control and rESC groups.CONCLUSIONThis study provided strong evidence to support that transplantation of rESC and ABMDN can improve functional recovery after iatrogenic SCI. The transplanted cells showed a beneficial therapeutic effect when compared to the control group.     

对侧皮质锁定螺钉与锁定螺钉治疗股骨远端骨折疗效比较

目的:比较对侧皮质锁定螺钉与锁定螺钉治疗股骨远端骨折的临床疗效。方法:回顾性分析自2013年5月至2016年8月诊治的52例股骨远端骨折患者,采用对侧皮质锁定螺钉+NCB接骨板内固定治疗26例(A组:对侧皮质锁定组),采用锁定螺钉+NCB接骨板内固定治疗26例(B组:锁定螺钉组)。记录两组手术出血量和手术时间、切口长度、内固定治疗后骨折愈合时间、内固定治疗后完全负重时间、内固定治疗后并发症发生率等,在每个随访节点对每位患者进行患肢的正侧位X线平片检查,末次随访时对患肢进行膝关节功能评分,采用美国特种外科医院膝关节评分标准评定患肢功能。骨折愈合的定义为活动时骨折处无痛且在骨折正侧位X线平片上可见到断端骨皮质骨痂连接。术后并发症包括:关节僵硬、内固定断裂、骨不连以及感染等。结果:本研究52例骨折均获得至少12个月的随访。两组在手术相关指标及切口愈合等方面均无明显差异(P均0.05)。在骨折愈合以及完全负重时间方面,A组均显著短于B组(P均0.05)。末次随访时52例患者患肢膝关节功能:A组:优18例,良5例,差4例,优良率88.5%;B组:优15例,良6例,中4例,差1例,优良率80.8%。两组对比A组优良率显著高于B组(P0.05)。两组并发症对比无明显差异:A组发生骨不连2例,骨折内固定断裂2例。B组发生骨不连3例,畸形愈合2例。结论:与传统锁定螺钉相比,对侧皮质锁定螺钉在骨折愈合时间、完全负重时间、术后患肢功能优良率方面具有优势,但在并发症发生率方面没有明显差异。对侧皮质锁定螺钉的治疗指征及自身强度还有待大样本、多中心的临床研究进一步明确。     

CGRP和NPY 在大鼠股骨闭合骨折愈合不同阶段的变化及意义

目的:探讨交感神经分泌的神经肽Y(NPY)和感觉神经分泌的钙基因相关肽(CGRP)在体内骨折愈合的不同阶段的变化及意义。方法:选择6-8月龄的雄性大鼠,建立大鼠的股骨闭合骨折模型,术后2、4、8、12周取材。进行扫描电镜,免疫组织荧光染色和血清Elisa检测。结果:1骨折愈合不同时期感觉神经肽类物质CGRP和交感神经肽类物质NPY都有表达,且其含量有先增加后减少的趋势,并在骨折后8周含量达到最高。2骨折愈合不同阶段的大鼠血清感觉神经肽类物质CGRP和交感神经肽类物质NPY均呈上升趋势,差异有统计学意义(P0.05),且NPY的含量比CGRP的含量高。骨折后2-4周,CGRP含量增加较快;骨折后4-8周NPY含量增加较快。结论:骨折愈合的不同阶段,感觉神经肽类物质CGRP和交感神经肽类物质NPY含量先升后降,对不同阶段的骨形成及骨吸收产生影响。     

血流变学指标及血清学指标在骨折延迟愈合患者中的临床意义

目的:探讨血流变学和血清学指标在骨折延迟愈合患者中的变化及其临床意义。方法:随机选取2010年1月~2016年6月在我院进行手术治疗的骨折延迟愈合及骨折正常愈合患者各90例,分别为观察组与对照组,对比分析两组患者术后第1、8、12周时血清可溶性血管细胞黏附分子1(sVCAM-1)、胰岛素样生长因子1(IGF-1)、血小板衍生生长因子(PDGF)及人可溶性细胞间黏附分子1(sICAM-1)和红细胞刚性指数、红细胞聚集指数、血浆黏度的差异。结果:术后第1、8、12周两组血清学及血流变学各指标整体相比差异均具有统计学意义(均P0.05),且组内两两比较均具有统计学差异(均P0.05)。术后8、12周观察组血清s ICAM-1、sVCAM-1、红细胞刚性指数、红细胞聚集指数、血浆黏度均高于对照组,而血清PDGF、IGF-1均低于对照组,比较差异均具有统计学意义(均P0.05)。结论:骨折患者血清sICAM-1、PDGF、IGF-1、sVCAM-1及红细胞刚性指数、红细胞聚集指数、血浆黏度水平会随着病程进展发生变化,并且血清sICAM-1、sVCAM-1及红细胞刚性指数、红细胞聚集指数、血浆黏度水平的升高,血清PDGF、IGF-1水平的降低可能是引起骨折延迟愈合的重要因素,对于骨折患者的临床治疗具有重要临床意义。     

Relationship between the Autoantibody and Expression of β3-Adrenoceptor in Lung and Heart

Background

Evidences suggest that β₃ -adrenoceptor (β₃-AR) plays an important role in heart failure (HF), although no data is reported indicating how these effects may change with the increasing age. Pulmonary congestion and edema are the major life-threatening complications associated with HF. The purpose of this study is to explore the relationship between the anti-β₃-AR autoantibody and the expression of β₃-AR in the lungs and heart for both aged patients and rats with HF.

Methods

Synthetic β₃-AR peptides served as the target antigens in ELISA were used to screen the anti-β₃-AR autoantibody in aged patients and rats. Two aged rat models were constructed based on aortic banding and sham-operation. The expression of β₃-AR mRNA and protein in the lung and heart was measured in intervention and non-intervention groups by Western blot analysis at the baseline, 5^th, 7^th, 9^th and 11^th week, respectively.

Results

The frequency and titer of anti-β₃-AR autoantibody in aged patients and rats with HF were higher than those in the control group (p<0.05). The expression of β₃-AR mRNA and protein in pulmonary tissues decreased continually from the 7^th week (p<0.05), followed by HF observed during the 9^th week. The expression of β₃-AR in myocardial tissues continued to increase after the 9^th week (p<0.05), and the expression of both β₃-AR mRNA and protein in the BRL group [HF group with BRL37344 (4-[-[2-hydroxy-(3-chlorophenyl)ethyl-amino] phenoxyacetic acid) (a β₃-AR agonist) injection] was positively correlated with BRL37344 when compared with non-BRL group (HF group without BRL37344 injection) (p<0.05).

Conclusion

Anti-β₃-AR autoantibody was detected in aged patients and rats with HF. The expression of β₃-AR mRNA and protein in pulmonary tissues decreased continually, and began earlier than in the heart, but its expression in myocardial tissues increased continually and could be further promoted by β₃-AR agonist.     

新型可降解锌合金内固定系统用于犬下颌骨骨折固定的研究

目的:探讨新型可降解锌合金内固定系统用于犬下颌骨骨折固定的可行性。方法:选择比格犬12只,并将其随机分为两组,其中6只为锌合金实验组,6只为钛合金对照组。每只比格犬的下颌作左右两个骨折内固定模型,每侧骨折模型使用一套小型四孔直连内固定产品。分别于术后即刻、术后4周、12周拍X片。于术前、术后4周、12周、24周抽血检测微量元素锌。于术后12周、24周分批次将动物安乐死,取下颌骨块行Micro-CT检测,取动物重要脏器和内固定周围软组织做病理切片。结果:全程动物无脱落,实验组与对照组骨折均临床愈合。X线图像及Micro-CT显示实验组与对照组的内固定效果比较差异无统计学意义(P0.05)。锌合金表面有明显的降解产物,Micro-CT测量锌合金的板钉初始体积为155.8±1.536 mm~3,12周体积为147.1±0.9893 mm~3,24周体积为137±5.365 mm~3,降解率12.07%,随着植入后时间的延长显著下降(P0.05),而钛合金板钉体积无显著变化。实验组及对照组心、肝、肾及植入物板周软组织均未见异常。血清锌离子浓度在正常范围内略有上升,各时间节点比较差异无统计学意义(P0.05)。结论:可降解锌合金内固定系统可以对犬下颌骨骨折提供稳固的固定,可降解锌合金在下颌骨部位具有一定的降解性能,血液中锌离子未见明显异常升高,对心、肝、肾及植入物周围软组织无毒性作用。     

强骨胶囊对老年股骨头近段骨折延迟愈合患者血清BMP-2 及IGF-1水平的影响

目的:探究强骨胶囊对老年股骨头近段骨折延迟愈合患者血清骨形态发生蛋白-2(BMP-2)及胰岛素生长因子-1(IGF-1)水平的影响。方法:选择我院收治的股骨近端骨折延迟愈合的老年患者41例,随机分为实验组及对照组。对照组19例予钙片;实验组22例予强骨胶囊。对比两组的临床疗效及治疗前后血清BMP-2及IGF-1水平的改变。结果:实验组总有效率(95.5%)高于对照组(78.9%),差异具备统计学意义(P0.05)。两组血清BMP-2及IGF-1水平均较治疗前显著升高(P0.05),且实验组血清BMP-2和IGF-1水平较对照组高(P0.05)。治疗后,两组血浆粘度均下降、骨密度值(BMD)均升高(P0.05);与对照组相较,实验组血浆粘度降低、BMD较高(P0.05)。结论:强骨胶囊能够有效改善老年股骨头近段骨折延迟愈合,促进骨折断端的愈合,推测其机制与增加患者血清BMP-2及IGF-1水平有关。     

白藜芦醇后处理对大鼠脑缺血再灌注损伤海马CA1区Bax、Bcl-2的影响

摘要 目的：探讨白藜芦醇后处理对大鼠脑缺血再灌注损伤Bax、Bcl-2表达的影响。方法：清洁级雄性SD大鼠60只随机分为假手术组（n=12）、I/R组（n=12）、白藜芦醇组（n=36）,白藜芦醇组按不同剂量分为低剂量、中剂量、高剂量组（10 mg/kg、20 mg/kg、40 mg/kg）,每组12只。假手术组：仅暴露大鼠颈外动脉,不做缺血处理;I/R组：采用改良线栓法制备大鼠大脑中动脉缺血再灌注损伤模型（缺血2 h,再灌注24 h）;白藜芦醇组：造模方法同I/R组,在大鼠缺血2h后,将不同剂量白藜芦醇腹腔注射入大鼠体内,比较各组SD大鼠神经功能缺损评分、采用Western blotting法、免疫组化法对大鼠脑组织缺血侧海马CA1区Bax和Bcl-2表达进行比较。结果：白藜芦醇低、中、高剂量组神经功能缺损评分均低于I/R组,随着白藜芦醇剂量的增加,神经功能缺损评分逐渐降低,其中白藜芦醇高剂量组神经功能缺损评分降低最为明显;白藜芦醇组与I/R组相比,不同剂量白藜芦醇组Bax表达逐渐减少,而Bcl-2表达明显增加,其中以白藜芦醇高剂量组改变最为明显。结论：高剂量白藜芦醇可以降低大鼠神经功能缺损评分,减轻脑缺血再灌注损伤,对大鼠脑缺血再灌注损伤具有保护作用,其机制与Bax、Bcl-2的表达有关。     

骨髓间充质干细胞移植对脑梗死大鼠神经功能恢复的影响

目的:观察骨髓间充质干细胞(BMSC)移植对脑梗死大鼠神经功能恢复的影响,并对其相关机制进行探讨。方法:90只大鼠随机分为3组:假手术组、对照组、BMSC移植组,每组30只。对照组和BMSC移植组建立大鼠大脑中动脉阻塞(MCAO)模型,假手术组只需要分离大鼠颈部组织,而不造MCAO模型。BMSC移植组在MCAO模型术后1天经尾静脉注射1 mL/3×10~6 BMSC,对照组注射同剂量的生理盐水,于MCAO术后1 d、3 d、7 d、14 d、21 d、28 d、35 d、42 d、49 d分别对各组大鼠进行神经功能评分(mNSS),术后2个月对BMSC移植组及对照组大鼠脑组织进行免疫组化染色,检测MAP2、TUJ1、Ⅷ因子、GFAP的表达情况。结果:在治疗后的第7天至第35天,BMSC移植组mNSS均显著低于对照组(P0.05)。术后2个月,BMSC移植组MAP2、TUJ1、Ⅷ因子表达量显著高于对照组,而GFAP表达量显著低于于BMSC对照组(P0.01)。结论:BMSC移植可以促进脑梗死神经功能的恢复。     

miR-126调控脑缺血大鼠细胞凋亡机制的研究

摘要 目的：探讨脑缺血大鼠microRNA-126（MiR-126）在脑组织和血浆中表达的动态变化及与细胞凋亡的关系。方法：健康雄性SD大鼠45只随机分为3组,其中3只大鼠不作任何处理,用于检测正常大鼠中MiR-126在脑组织和血浆的表达,剩余大鼠随机分为假手术组和模型组,模型组用线栓法造脑缺血大鼠模型。TTC法检测模型组大鼠脑组织梗死病变用以确定造模是否成功;RT-qPCR检测大鼠造模后1、3、6、9、12、24和48小时脑组织和血浆中MiR-126的动态表达变化;采用苏木精-伊红染色法检测脑组织形态学变化;采用Western blot 方法分析相关凋亡蛋白表达的变化。结果：TTC法和H-E染色结果表明,大鼠脑缺血造模成功,模型组大鼠出现脑组织大面积梗死病变;MiR-126和Caspase-3表达在模型组大鼠脑组织和血浆中随着脑缺血时间的延长其表达水平逐渐提高,24 h到达高峰后,在48 h Mi-R126表达强度又降低,但仍然高于假手术组大鼠水平;Caspase-3在正常对照组和假手术组表达量都较少（P>0.05）,其他凋亡蛋白Bax和 STAT3相对表达水平趋势同 Caspase-3相似;而Bcl-2表达水平明显降低,趋势与其他凋亡蛋白相反。结论：MiR-126在脑缺血大鼠脑组织和血浆中表达水平明显升高,而且细胞凋亡明显增加,因此MiR-126可能通过促进细胞凋亡对脑缺血大鼠有保护作用,此研究为脑缺血发病机制和诊断提供更多依据。     

Celecoxib Ameliorates Non-Alcoholic Steatohepatitis in Type 2 Diabetic Rats via Suppression of the Non-Canonical Wnt Signaling Pathway Expression

Our aim was to test whether pharmacological inhibition of cycloxygenase-2 (COX-2) reverses non-alcoholic steatohepatitis (NASH) in type 2 diabetes mellitus (T2DM) rats via suppression of the non-canonical Wnt signaling pathway expression. Twenty-four male Sprague-Dawley rats were randomly distributed to two groups and were fed with a high fat and sucrose (HF-HS) diet or a normal chow diet, respectively. After four weeks, rats fed with a HF-HS diet were made diabetic with low-dose streptozotocin. At the 9^th week the diabetic rats fed with a HF-HS diet or the non-diabetic rats fed with a normal chow diet were further divided into two subgroups treated with vehicle or celecoxib (a selective COX-2 inhibitor, 10 mg/Kg/day, gavage) for the last 4 weeks, respectively. At the end of the 12^th week, rats were anesthetized. NASH was assessed by histology. Related cytokine expression was measured at both the protein and gene levels through immunohistochemistry (IHC), Western blot and real-time PCR. T2DM rats fed with a HF-HS diet developed steatohepatitis and insulin resistance associated with elevated serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), insulin levels and the non-alcoholic fatty liver disease (NAFLD) activity score (NAS). The expression of Wnt5a, JNK1, NF-κB p65, and COX-2 were all significantly increased in the T2DM-NASH group compared with the control and control-cele group. Hepatic injury was improved by celecoxib in T2DM-NASH-Cele group indicated by reduced serum ALT and AST levels and hepatic inflammation was reduced by celecoxib showed by histology and the NAFLD activity score (NAS). Serum related metabolic parameters, HOMA-IR and insulin sensitivity index were all improved by celecoxib. The expression of Wnt5a, JNK1, NF-κB p65, and COX-2 expression were all suppressed by celecoxib in T2DM-NASH-Cele group. The results of the present study indicated that celecoxib ameliorated NASH in T2DM rats via suppression of the non-canonical Wnt5a/JNK1 signaling pathway expression.     

续断对兔骨折愈合过程中相关基因表达和血钙、磷含量的影响

本研究通过检测中药材续断对家兔骨折模型愈合过程中与成骨密切相关基因的表达,以及血清中钙、磷含量变化,探讨其对骨折愈合的促进机制。构建家兔骨折缺损模型,术后按组分别给予续断和蒸馏水灌胃,并分别检测骨保护素(OPG)、骨保护素配体(OPGL)、局部转化生长因子β1(TGF-β1)和骨形态发生蛋白-2(BMP-2)的基因表达以及血清中Ca、P、碱性磷酸酶(ALP)含量。结果表明,续断治疗组中血钙、血磷和ALP的含量在灌胃第2周,第3周和第4周后均有明显升高,且在第三周时达到最大值。同时,OPG、TGF-β1、BMP-2三个基因在用续断治疗的不同时期呈现不同程度的表达上调,OPGL则在治疗早期表达下调。推测续断对骨折的治疗可能是通过调控OPG、OPGL、TGF-β1、BMP-2等基因在骨愈合不同阶段的表达量和血清中Ca、P、ALP的含量来促进骨骼生长。     

Lentivirus-mediated Wnt10b overexpression enhances fracture healing in a rat atrophic non-union model

Lentivirus vectors encoding Wnt10b gene (LV–Wnt10b) or luciferase gene (LV-luc) were constructed to determine whether Wnt10b overexpression improved fracture healing in a rat atrophic non-union model. After fracture, rats were injected with LV-Wnt10b or LV-luc. Luciferase signals were clearly detected. At 2 and 4 weeks, LV-Wnt10b group had 107 and 98 % more proliferating cell nuclear antigen (PCNA) positive cells, respectively, and promoted expression of bone morphogenetic protein-2 (BMP-2) in the callus compared with controls. LV-Wnt10b injection significantly increased bone mass density and bone mineral content: 46–84 and 96–193 %, respectively, at the site of fracturein the LV-Wnt10b group compared with controls. At 8 weeks, fractured femora were healed in the LV-Wnt10b group compared with atrophic non-unions formed in controls. Thus, Wnt10b overexpression associated with lentiviral gene therapy is effective in healing atrophic non-unions in rats.