体外培养骨髓单个核细胞分泌促血管生长因子的实验研究

目的分析体外培养的骨髓单个核细胞持续分泌促血管生长因子的能力。方法从大鼠胫骨及股骨采集骨髓,密度梯度离心法分离出骨髓单个核细胞进行体外培养,并连续收集4周培养上清液。酶联免疫吸附实验(ELISA)法测定培养上清液中碱性成纤维细胞生长因子(bFGF)、血管内皮细胞生长因子(VEGF)和白介素-1β(IL-1β)等因子水平。结果第1、2、3、4周骨髓单个核细胞体外培养上清液中VEGF分别为(24.40±7.99)pg/m、l(89.28±5.13)pg/m、l(115.24±10.08)pg/m、l(157.00±15.64)pg/m l;bFGF含量分别为(52.72±2.13)pg/m、l(48.10±6.41)pg/m、l(44.71±3.21)pg/m、l(25.61±2.42)pg/m l;IL-1β含量分别为:(31.28±5.44)pg/m、l(71.87±3.01)pg/m、l(55.77±11.94)pg/m、l(41.75±9.14)pg/m。l结论体外培养骨髓单个核细胞可持续分泌VEGF、bFGF、IL-1β等多种促血管生长因子。     

Ex Vivo Application of Carbon Monoxide in University of Wisconsin Solution to Prevent Intestinal Cold Ischemia/Reperfusion Injury

Carbon monoxide (CO), a byproduct of heme catalysis, was shown to have potent cytoprotective and anti-inflammatory effects. In vivo recipient CO inhalation at low concentrations prevented ischemia/reperfusion (I/R) injury associated with small intestinal transplantation (SITx). This study examined whether ex vivo delivery of CO in University of Wisconsin (UW) solution could ameliorate intestinal I/R injury. Orthotopic syngenic SITx was performed in Lewis rats after 6 h cold preservation in control UW or UW that was bubbled with CO gas (0.1-5%) (CO-UW). Recipient survival with intestinal grafts preserved in 5%, but not 0.1%, CO-UW improved to 86.7% (13/15) from 53% (9/17) with control UW. At 3 h after SITx, grafts stored in 5% CO-UW showed improved intestinal barrier function, less mucosal denudation and reduced inflammatory mediator upregulation compared to those in control UW. Preservation in CO-UW associated with reduced vascular resistance (end preservation), increased graft cyclic guanosine monophosphate levels (1 h), and improved graft blood flow (1 h). Protective effects of CO-UW were reversed by ODQ, an inhibitor of soluble guanylyl cyclase. In vitro culture experiment also showed better preservation of vascular endothelial cells with CO-UW. The study suggests that ex vivo CO delivery into UW solution would be a simple and innovative therapeutic strategy to prevent transplant-induced I/R injury.     

移植人羊膜细胞对大鼠创伤性脑损伤的实验研究

目的 探究大鼠TBI后脑内移植人羊膜细胞（HACs）对大鼠运动功能的影响。方法 HACs经分离、Hoechst33342标记后重悬调整细胞浓度为10^5/μl；采用改进的Feeney自由落体法打击大鼠脑皮层后肢运动区域，损伤后24h经微量注射器和立体定向仪将Hoechst33342标记的HACs 10μl分别移植于挫伤灶中心和挫伤灶边缘；在TBI后的28d内采用钉板平衡木行走测试大鼠运动功能变化，运动功能检测结束后取出脑组织行组织学检测。结果 治疗组滑落脚步数明显少于对照组（P〈0．05）；移植的HACs呈蓝色荧光；部分移植HACs可见MAP-2阳性表达。结论 移植HACs使大鼠TBI后运动功能明显改善。     

Particulate endocytosis mediates biological responses of human mesenchymal stem cells to titanium wear debris.

Continual loading and articulation cycles undergone by metallic (e.g., titanium) alloy arthroplasty prostheses lead to liberation of a large number of metallic debris particulates, which have long been implicated as a primary cause of periprosthetic osteolysis and postarthroplasty aseptic implant loosening. Long-term stability of total joint replacement prostheses relies on proper integration between implant biomaterial and osseous tissue, and factors that interfere with this integration are likely to cause osteolysis. Because multipotent mesenchymal stem cells (MSCs) located adjacent to the implant have an osteoprogenitor function and are critical contributors to osseous tissue integrity, when their functions or activities are compromised, osteolysis will most likely occur. To date, it is not certain or sufficiently confirmed whether MSCs endocytose titanium particles, and if so, whether particulate endocytosis has any effect on cellular responses to wear debris. This study seeks to clarify the phenomenon of titanium endocytosis by human MSCs (hMSCs), and investigates the influence of endocytosis on their activities. hMSCs incubated with commercially pure titanium particles exhibited internalized particles, as observed by scanning electron microscopy and confocal laser scanning microscopy, with time-dependent reduction in the number of extracellular particles. Particulate endocytosis was associated with reduced rates of cellular proliferation and cell-substrate adhesion, suppressed osteogenic differentiation, and increased rate of apoptosis. These cellular effects of exposure to titanium particles were reduced when endocytosis was inhibited by treatment with cytochalasin D, and no significant effect was seen when hMSCs were treated only with conditioned medium obtained from particulate-treated cells. These findings strongly suggest that the biological responses of hMSCs to wear debris are triggered primarily by the direct endocytosis of titanium particulates, and not mediated by secreted soluble factors. In this manner, therapeutical approaches that suppress particle endocytosis could reduce the bioreactivity of hMSCs to particulates, and enhance long-term orthopedic implant prognosis by minimizing wear-debris periprosthethic osteolysis.     

直肠癌切除前后腹腔冲洗液脱落细胞学对比研究

目的研究直肠癌术中腹腔多次冲洗，对于降低患者腹腔内脱落癌细胞阳性率的临床意义。方法对63例直肠癌患者术中腹腔冲洗分6次进行，肿瘤切除前3次，切除后3次；将6次腹腔冲洗液行常规病理学涂片细胞学检查结果进行对比。结果肿瘤切除前，全组患者腹腔冲洗液脱落癌细胞均呈阳性表达。肿瘤切除后第1次腹腔冲洗液脱落癌细胞阳性40例，第2次阳性33例，第3次阳性13例。肿瘤切除后第1次冲洗液的脱落癌细胞阳性结果与切除后第2次冲洗液阳性结果比较，P〉0.05，差异无统计学意义；但第3次冲洗液的脱落癌细胞阳性结果与第2次比较，P〈0.01，差异有统计学意义。结论对直肠癌患者进行术中多次腹腔冲洗可降低腹腔内脱落癌细胞的阳性率。     

血管内皮生长因子及炎性因子在油酸性犬急性呼吸窘迫综合征中的变化

目的探讨油酸性急性呼吸窘迫综合征（ARDS）beagle犬血浆及肺泡灌洗液血管内皮生长因子（VEGF）、可溶性细胞间黏附分子-1（ICAM-1）、白细胞介素-8（IL-8）、肿瘤坏死因子-α（TNF—α）水平的改变。方法12只英国纯种beagle犬，静脉注射油酸0．15mL／kg，在注射油酸前、后1h，出现ARDS的典型表现时，抽血测VEGF、sICAM-1、IL-8、TNF—α，并对此时相作肺泡灌洗液VEGF、sICAM—1、IL-8、TNF-α的测定。结果beagle犬静脉注射油酸后1h血浆TNF—α升高（P〈0．05），血浆反肺泡灌洗液IL8、sICAM-1和VEGF在1h较油酸前没有明显变化（P〉0．05），beagle犬油酸型ARDS模型建立后血浆及肺泡灌洗液VECF、sICAM-1、IL-8、TNF—α均显著高于建模前（P〈0．05）。结论VEGF、sICAM-1、IL-8、TNF～α在beagle犬油酸型ARDS发生发展过程中可能均起重要作用，其水平的高低可能与ARDS严重程度及预后有关。     

Gastrointestinal stromal tumours: a regular origin in the muscularis propria, but an extremely diverse gross presentation

Background Gastrointestinal stromal tumours (GISTs) are thought to arise from the interstitial cells of Cajal (ICCs). ICCs form a network surrounding the myenteric plexus and between-muscle fibres of the muscularis propria of the tubular GI tract. The cell of origin of so-called extra-gastrointestinal stromal tumours (EGISTs) is not known.Aim and methods To study the diversity of gross presentation of GISTs and to critically assess the incidence of EGISTs and their relationship to mural GISTs, a total of 200 neoplasms with typical morphologic and immunohistochemical features of GISTs were reviewed, looking for any degree of association with the muscularis propria of the gut wall.Results There were 130 gastric (65%), 9 duodenal (4.5%), 48 small intestinal (24%), 9 colorectal (4.5%), 1 appendiceal (0.5%) and 3 unclassifiable GISTs (1.5%). Fourteen cases (7%) were initially submitted as EGISTs (four mesenteric, four omental, one pararectal/prostatic, one pelvic/Douglas, one perivesical, one located between root of mesentery and tail of pancreas, one involving the mesentery, omentum and abdominal wall extensively and one located between liver and stomach). After critical re-evaluation of surgical reports and remote clinical history and a careful search for residual muscular tissue from the gut wall in the tumour pseudocapsule (in some cases supported by desmin immunoreactivity), it was possible to reclassify most of these cases (11/14) as either GISTs with extensive extramural growth resulting in loss of contact to the external muscle coat of the gut (8/14) or as metastases from an inoperable GIST (2/14) or from a previously resected deceptively benign tumour (1/14).Conclusion EGISTs are probably rarer than previously reported (1.5% or less in this study). We concluded that most so-called EGISTs represent apparent EGISTs that should have arisen from the outermost muscle coat, but have lost their contact to the point of origin due to extensive extramural growth pattern. From a surgical point of view, it is crucial to document and mark any focal attachment or adhesions to the gut wall noticed during surgery for an apparent EGIST. In contrast to most other neoplasms, GISTs should be defined by virtue of any degree of association with the muscularis propria (no matter how minimal), but not by localisation of the bulk of the tumour.