Osteogenic potential of postnatal skeletal muscle-derived stem cells is influenced by donor sex.

This study compared the osteogenic differentiation of F-MDSCs and M-MDSCs. Interestingly, M-MDSCs expressed osteogenic markers and underwent mineralization more readily than F-MDSCs; a characteristic likely caused by more osteoprogenitor cells within the M-MDSCs than the F-MDSCs and/or an accelerated osteogenic differentiation of M-MDSCs. INTRODUCTION: Although therapies involving stem cells will require both female and male cells, few studies have investigated whether sex-related differences exist in their osteogenic potential. Here, we compared the osteogenic differentiation of female and male mouse skeletal muscle-derived stem cells (F- and M-MDSCs, respectively), a potential cell source for orthopedic tissue engineering. MATERIALS AND METHODS: F- and M-MDSCs were stimulated with bone morphogenetic protein (BMP)4, followed by quantification of alkaline phosphatase (ALP) activity and expression of osteogenic genes. F- and M-MDSCs were also cultured as pellets in osteogenic medium to evaluate mineralization. Single cell-derived colonies of F- and M-MDSCs were stimulated with BMP4, stained for ALP, and scored as either Low ALP+ or High ALP+ to detect the presence of osteoprogenitor cells. F- and M-MDSCs were transduced with a BMP4 retrovirus (MDSC-BMP4 cells) and used for the pellet culture and single cell-derived colony formation assays. As well, F- and M-MDSC-BMP4 cells were implanted in the intramuscular pocket of sex-matched and sex-mismatched hosts, and bone formation was monitored radiographically. RESULTS AND CONCLUSIONS: When stimulated with BMP4, both F- and M-MDSCs underwent osteogenic differentiation, although M-MDSCs had a significantly greater ALP activity and a larger increase in the expression of osteogenic genes than F-MDSCs. In the pellet culture assay, M-MDSCs showed greater mineralization than F-MDSCs. BMP4 stimulation of single cell-derived colonies from M-MDSCs showed higher levels of ALP than those from F-MDSCs. Similar results were obtained with the MDSC-BMP4 cells. In vivo, F-MDSC-BMP4 cells displayed variability in bone area and density, whereas M-MDSC-BMP4 cells showed a more consistent and denser ectopic bone formation. More bone formation was also seen in male hosts compared with female hosts, regardless of the sex of the implanted cells. These results suggest that M-MDSCs may contain more osteoprogenitor cells than F-MDSCs, which may have implications in the development of cellular therapies for bone healing.     

转化生长因子β1短发夹RNA对白蛋白刺激人肾近曲小管上皮细胞细胞因子表达的影响

目的 研究pcDU6载体质粒介导的转化生长因子β1（TGF-β1）短发夹RNA（shRNA）对人血清白蛋白（HSA）致人肾近曲小管上皮细胞（HK2细胞）增殖，TGF-β1、结缔组织生长因子（CTGF）和纤连蛋白（FN）表达的影响，并探讨有关机制。方法 构建TGF-β1shRNA的pcDU6载体质粒，体外培养HK2细胞株。采用脂质体转染将表达TGF-β1shRNA的pcDU6质粒载体（pcDU6-A1-A2和peDU6-B1-B2）分别导人实验组细胞。用HSA（5g／L）刺激HK2细胞12h或24h。四甲基偶氮唑盐（MTF）比色法测定细胞增殖水平。RT-PCR半定量分析HK2细胞中TGF-β1、CTGF和FNmRNA的表达水平；双抗夹心酶联免疫吸附法检测HK2细胞培养液中TGF-β1及FN蛋白质水平。结果 HK2细胞在HSA刺激下，其TGF-β1、CTGF及FNmRNA的表达明显上调，培养液中TGF-β1和FN的蛋白质含量亦明显升高（P〈0．05）。与pcDU6空载体组比较，pcDU6载体质粒介导的TGF-β1shRNA干扰组TGF-β1、CTGF及FNmRNA的表达明显下调（P〈0．05）。TGF-β1shRNA转染HK2细胞后12h或24h，细胞培养液中TGF-β1和FN蛋白质含量明显下降，HK2细胞增殖被部分抑制（P〈0．05）。在细胞增殖、TGF-β1、CTGF及FN基因表达方面，TGF-β1shRNA干扰组组间比较，以及pcDU6空载体转染组与HSA刺激组比较，差异均无统计学意义。结论 peDU6载体质粒介导的TGF-β1shRNA能够明显抑制HSA刺激下HK2细胞增殖、TGF-β1、CTGF和FN基因的表达。HSA刺激HK2细胞增殖及CTGF和FN基因的过表达可能通过TGF-β1介导。     

明胶降解物对大鼠真皮成纤维细胞增殖及Ⅰ型胶原蛋白分泌的影响

目的 探讨组织工程心瓣膜常用人工基质材料--明胶发生降解后的产物对大鼠真皮成纤维细胞增殖及Ⅰ型胶原蛋白分泌的影响. 方法 利用酶组合技术制备明胶降解物（gelatin hydrolysate, GH）,采用超滤的方法分离出低分子量明胶降解物组分（gelatin hydrolysate fraction, GHF）,用于5只SD大鼠真皮成纤维细胞的体外培养.采用CCK-8活细胞计数试剂盒（Cell Counting Kit-8）检测细胞的增殖情况,ELISA试剂盒检测细胞I型胶原蛋白的分泌情况. 结果 ①GHF对细胞增殖作用的浓度效应：在0.125～1.0 mg/ml浓度范围内,GHF对细胞的促增殖作用呈现先增强后减弱的趋势,在0.25 mg/ml时,细胞增殖率最大,达到（47.54±16.35）%;②GH与GHF对细胞增殖影响比较：GH与GHF均具有促进细胞增殖的作用,且在浓度为0.25 mg/ml时,促细胞增殖作用无显著差异[（27.04±15.21）% vs （39.22±15.55）%, P=0.177];③GH与GHF对细胞分泌Ⅰ型胶原蛋白影响比较：GHF组第3天时Ⅰ型胶原分泌量为（28.06±5.60）pg/105,与空白对照的（18.24±4.52）pg/105相比,有显著差异（P=0.006）,而GH组直至第6天时Ⅰ型胶原蛋白分泌量,与空白对照相比,无统计学意义（P=0.103）. 结论 分子量大小不同的明胶降解物GH与GHF均具有促进大鼠真皮成纤维细胞增殖的作用,且分子量较小的GHF还具有促进细胞I型胶原分泌的功能.     

义齿PMMA基托表面粗糙度影响的实验研究

目的：研究技工操作因素对义齿树脂基托表面粗糙度的影响。方法：在不同粉液比、充填时期、聚合温度、混合方式以及出盒温度成型的上颌半口义齿基托上选取1.5cm^2范围，按照统一的打磨顺序，由粗到细打磨抛光完成，浸入冷水中，7天后进行粗糙度测试，所得数据经t检验统计处理。结果：在单体含量降低组、丝状期充填组．热处理时直接放入70℃水中维持9h组与对照组相比，均存在显著差异（P〈0．05）。结论：影响PMMA树脂表面粗糙度的制作工艺因素有单体含量降低、丝状期充填和未经100℃高温热处理。     

心包膜移植在结膜囊成形术中的应用

目的：探讨保存人心包膜移植治疗结膜囊狭窄的临床效果及安全性。方法：对眼球摘除术后伴有结膜囊狭窄综合征的26例26眼，利用心包膜移植行部分结膜囊成形术，术后随访观察3～12个月，平均8个月。结果：24眼（92．31％）在术后30～60天移植的心包膜被结膜覆盖，感染1例（3．85％），结膜囊狭窄1例（3．85％）。结论：保存心包膜移植治疗结膜囊狭窄取材方便，疗效显著，操作简单。     

F-Z液与HC-A液对内皮细胞保存效果的比较研究

HC-A液（高渗枸橼酸盐腺嘌呤液）是由第二军医大学附属长征医院（何长民等）与上海市中心血站（刘书元等）在Ross溶液基础上加以改良而成，经过约20年的肾移植临床应用，获得广泛的肯定。但HC-A液对于肝脏、胰腺和心脏的保存效果较差，不能满足临床需要，因此，笔者对其进行改进，并取得了较好的结果，命名为F-Z液，下文就是在内皮细胞水平，对两者的保存效果进行比较的结果。[第一段]     

脂膜微泡结合凝血酶原复合物的制备

目的:制备结合凝血酶原复合物(PCC)的阴离子脂膜微泡,评价微泡的理化性质。材料和方法:采用机械振荡直接连接法,分两组(于机械振荡之前或之后加入),制备结合PCC的阴离子脂膜微泡,观察并检测洗涤前后微泡的理化性质。结果:微泡与PCC的结合率及Ⅸ因子活性:两组间比较无明显差异(P>0.05),但洗涤后结合率由洗涤前的95%降为80%,活性由90%降为19%。结论:直接连接法可制备结合PCC的阴离子脂膜微泡,微泡与PCC结合率较高,且洗涤前能保持Ⅸ因子较高的活性。     

关节镜下人工韧带重建前交叉韧带的临床初步体会

目的探讨关节镜下应用LARS人工韧带重建前交叉韧带的可行性及近期疗效. 方法用法国产LARS人工韧带对16例前交叉韧带(anterior cruciate ligament,ACL)损伤行关节镜下ACL重建术.等距点钻胫骨、股骨骨道,将肌腱拉入骨道,韧带游离部分位于关节腔内,拉紧后2枚螺钉固定韧带,合并损伤同期处理. 结果手术时间51～86 min,平均64 min.术后无滑膜炎、韧带断裂、活动明显受限等并发症.16例均随访1.5～6个月,平均3.8月.按照IKDC评分标准:术前C级6例,D级10例;术后A级6例,B级9例, C级1例(χ2=6.264,P<0.05).Lysholm膝关节功能评分术前36～76分,(63.7±7.3)分;术后86～97分,(94.8±9.6)分(t=10.356,P<0.05). 结论关节镜下LARS人工韧带重建ACL,操作简便,可使膝关节获得即时稳定性,早期康复锻炼,最大限度的防止关节功能受限,近期疗效满意.     

超声对胰腺移植术后的监测作用

目的探讨二维超声和彩色多普勒血流显像（cDFI）对移植胰腺的功能和术后并发症的监测作用。方法用彩色多普勒超声观察6例移植胰腺的形态和血流动力学变化情况。结果（1）功能正常的移植胰腺在术后形态正常，体积偏大，主胰管内径正常，实质回声较正常胰腺略高，动脉PSV13～64cm/s，R10.44～0.68，P10.67～2.33。（2）2例出现急性排异和1例出现移植胰腺炎的移植胰腺周边出现低或无回声区，实质回声明显减低，分布不均。1例急性重度排异的RI值超过0.9，而另1例急性轻度排异RI值小于0.7。1例移植胰腺炎RI值超过0.9，PI值超过4.0。结论CDFI作为一种无创的检查技术对胰腺移植术后的监测和诊治具有积极意义。     

Protein adsorption on ex vivo catheters and polymers exposed to peritoneal dialysis effluent.

BACKGROUND: Deposition of proteins on surfaces of medical devices has been recognized to putatively relate to the process of regulation of biomaterial-associated complications by attachment of fibrin clots, eukaryotic cells, and microbes. The molecules adsorb to a varying extent, depending not only on the physicochemical properties of the biomaterial, but also on the composition of the host fluid. OBJECTIVE: Adsorption of proteins on catheters exposed both ex vivo and in vitro to dialysate of patients on peritoneal dialysis (PD) was studied. METHODS: Peritoneal dialysis effluent was collected from 5 patients with end-stage renal disease on continuous ambulatory PD. Tenckhoff catheters were obtained from 16 patients. Deposition of proteins on excised Tenckhoff catheters and tubing of different materials exposed to PD effluent in vitro was studied using 125iodine-labeled antibodies. Adhesion of Staphylococcus aureus and Staphylococcus epidermidis strains was quantified on tubing exposed to PD effluent in vitro. RESULTS: The presence of albumin, transferrin, immunoglobulin G, fibrinogen, fibronectin, von Willebrand factor, vitronectin, and thrombospondin was determined at various concentrations in PD effluent. All proteins analyzed were detected on PD catheters removed from patients. The extent of protein deposition on Tenckhoff catheters exposed to PD effluent, in vitro, rapidly reached a plateau and remained constant, as it did on polyvinyl chloride and polyethylene tubing. Adhesion of staphylococci was enhanced on Tenckhoff catheters exposed to PD effluent compared to unused PD solution. CONCLUSIONS: The data identify surface exposed proteins that may serve as adhesion sites for microbes on peritoneal catheters indwelled in patients undergoing PD.