小肠粘膜下层的制备及细胞相容性的实验研究

目的了解猪小肠粘膜下层(SIS)的细胞相容性,探讨用SIS为生长载体复合骨髓基质干细胞(BMSCs)构筑组织工程骨的可能性。方法用物理和化学方法处理猪小肠粘膜下层,将兔骨髓基质干细胞与SIS进行体外复合培养,分别进行组织学、相差显微镜和扫描电镜观察。结果经物理和化学处理的SIS纯度高,孔隙多,胶原纤维未受损;BMSCs在SIS材料上生长、粘附、增殖,并能长入材料的孔隙内,分泌大量的细胞外基质成分。结论SIS的细胞相容性良好,不影响BMSCs的形态,对细胞生长和功能表达无抑制作用,可以用作骨组织工程的支架材料。     

小肠粘膜下层复合成骨诱导的骨髓基质干细胞体内成骨的实验研究

目的探讨利用组织工程方法,以小肠粘膜下层为支架材料复合成骨诱导后的骨髓基质干细胞构建骨组织的可行性。方法将取自兔骨髓中的骨髓基质干细胞经成骨诱导液诱导后,与经处理的猪小肠粘膜下层在体外共培养。1周后,将共培养的猪小肠粘膜下层埋置于无胸腺裸鼠皮下。分别在不同时间进行扫描电镜、透射电镜、组织学和免疫组织化学观察。结果体外培养时,见细胞与材料粘附良好,且分泌大量的细胞外基质,细胞分化、增殖活跃。大体观察植入体内的细胞-材料复合物,见颜色变白,组织硬度增加,组织学和电镜观察见有大量骨组织形成。免疫组化示细胞为具有分泌特异性骨钙蛋白的成骨细胞。结论骨髓基质干细胞经成骨诱导为成骨细胞后与小肠粘膜下层共培养,植入裸鼠体内后可形成骨组织,小肠粘膜下层是一种良好的骨组织工程支架材料。     

骨髓基质干细胞复合小肠黏膜下层体外构建组织工程骨膜的实验研究

目的 探讨用猪小肠粘膜下层（SIS）作支架,复合骨髓问充质干细胞（BMSC）体外构建组织工程骨膜的可行性。方法用常规方法培养BMSC,再与SIS进行体外复合培养,分别进行相差显微镜、透射电镜、扫描电镜和组织学检查。观察BMSC在SIS上的生长、分化、增殖及细胞分泌细胞外基质情况。结果BMSC在SIS材料上粘附、增殖,分泌大量的细胞外基质成份,细胞功能活跃,在SIS上成多层生长,厚度随复合培养时间的延长而增加,类似生物骨膜。结论SIS与BMSC在体外复合培养可以构建出类似生物骨膜的组织工程骨膜,为进一步研究体内成骨奠定基础。     

冻存对组织工程骨膜生物活性的影响

目的 观察冻存复苏过程对组织工程骨膜生物学特性的影响.方法 体外培养新西兰大白兔骨髓间充质干细胞(BMSCs),与猪小肠黏膜下层(SIS)构建组织工程骨膜.冻存后复苏培养,扫描电镜(SEM)观查复苏后种子细胞生长状况、噻唑蓝(MTT)比色法绘生长曲线、钙-钴法检测碱性磷酸酶(ALP)、生物化学法定量分析ALP的表达.结果 冻存复苏后BMSCs仍能在SIS上保持良好的生长状况;组织工程骨膜复苏后成骨诱导培养5d时ALP含量达峰值,10～15d时稳定在较低的量,与对照组比较差异有统计学意义(P<0.05). 结论组织工程骨膜冻存复苏后仍保持较稳定的生物活性.     

不同年龄人骨髓间充质干细胞体外增殖及成骨分化的研究

目的 :观测年龄对人骨髓间充质干细胞 (humanmesenchymalstemcells ,hMSCs)体外增殖、成骨分化的影响。方法 :使用密度梯度离心法分离不同年龄人骨髓MSCs进行培养 ,保留贴壁细胞传代 ,观察细胞生长情况 ,检测其增殖活性、碱性磷酸酶活性 (ALP)、诱导后骨钙素定量测定。结果 :低龄hMSCs较高龄hMSCs体外生长快、MTT、ALP及骨钙素浓度高。结论 :hMSCs的增殖和成骨分化的能力和活性随着年龄的增加而降低。     

骨髓间充质干细胞与骨组织共培养模型的建立

目的:建立骨髓间充质干细胞(BMSCs)与骨组织共培养模型,模拟体内成骨环境,以全面研究骨髓间充质干细胞及细胞支架复合物的生物性状.方法:通过插入式培养皿和培养板来构建细胞一组织共培养的模型,将新鲜兔骨碎粒及骨块与骨髓间充质干细胞共培养,观察共培养条件下细胞的形态学变化、ALP活性及矿化能力,免疫组化检测Ⅰ型胶原、骨钙素.结果:构建出骨髓间充质干细胞与骨组织共培养的模型.共培养的间充质干细胞同期ALP活性高于普通培养对照组,其Ⅰ型胶原、骨钙素免疫组化阳性,对照组Ⅰ型胶原免疫组化弱阳性、骨钙素免疫组化阴性.结论:体外构建的共培养模型部分模拟了体内成骨环境;经过共培养的细胞呈现成骨细胞的表型.     

骨髓间充质干细胞复合组织工程化脱细胞真皮基质构建组织工程皮肤

目的：体外培养扩增SD大鼠骨髓间充质干细胞（bone marrow mesenchymal stem cells,BMSCs），复合组织工程化脱细胞真皮基质构建组织工程皮肤，为进一步临床应用奠定基础。方法：将SD大鼠骨髓间充质干细胞进行体外培养扩增后，以生长状态良好的骨髓间充质干细胞接种于制备好的组织工程化脱细胞真皮支架上，进行体外联合培养，构建组织工程皮肤。观察细胞生长情况及组织工程皮肤结构。结果：体外培养的SD大鼠骨髓间充质干细胞生长良好，传代扩增容易，组织工程化脱细胞真皮基质去细胞完全，骨髓间充质干细胞在脱细胞真皮基质中生长良好，可体外构建组织工程皮肤。结论：利用体外扩增培养的骨髓间充质干细胞及制备的组织工程化脱细胞真皮基质可以体外联合构建组织工程皮肤。     

骨髓基质干细胞复合小肠黏膜下层体内异位成骨的研究

目的 观察骨髓间充质干细胞复合小肠黏膜下层构建组织工程骨膜的异位成骨能力.方法 将常规方法培养的BMSCs与SIS复合(M1组)及进行成骨诱导培养BMSCs与SIS复合(M2组)植入兔(n=8)背部肌袋内作为实验组,以单纯SIS作为阴性对照.术后4、8、12周观察其成骨情况.结果 BMSCs在SIS支架上黏附、生长良好.植入体内4、8、12周,M2、M1、SIS组成骨量分别为(25.08±0.79)%、(18.81±0.42)%和(13.98±1.86)%,各指标M2组高于M1或SIS组(P<0.05),M1组高于SIS组(P<0.05).随着时间的延长,两实验组成骨量逐渐增加(P<0.05),SIS组的成骨量变化不明显(P>0.05). 结论 BMSCs与SIS复合构建组织工程骨膜在体内有异位成骨作用,而且经过成骨诱导的组织工程骨膜成骨作用更加明显.     

人间充质干细胞在骨组织工程中的应用

目的从数量与功能两方面探讨人间充质干细胞(humanmesenchymalstemcells,hMSCs)作为骨组织工程种子细胞的可行性。方法髂骨穿刺,梯度离心分离获得hMSCs,体外扩增培养;对第3代细胞用地塞米松、β-甘油磷酸、维生素C等成骨诱导培养,检测细胞碱性磷酸酶、骨钙素的表达和钙结节形成情况;诱导细胞与可吸收性复合支架材料体外构建复合体,植入裸小鼠皮下,对照组仅植入支架材料,术后3周取材检测骨钙素、Ⅰ型胶原表达。结果4ml骨髓含3.2×107～6.0×107个单个核细胞,培养3周收获hMSCs2.5×107～4.3×107;成骨诱导后,表达碱性磷酸酶、骨钙素阳性细胞数>50%,对照组<10%;诱导细胞与材料复合后植入体内仍然高表达骨钙素、Ⅰ型胶原。结论hMSCs能够满足体外构建组织工程化骨,对种子细胞数量和功能的要求。     

联合培养时诱导的内皮细胞对自体骨髓基质干细胞成骨作用的影响

目的探讨由骨髓基质干细胞(BMSCs)诱导的内皮细胞(EC)与自体BMSCs共培养时对BMSCs成骨作用的影响。方法采用密度梯度离心法分离兔BMSCs,培养细胞分为四组：A组(BMSCs组)、B组(BMSCs成骨诱导组)、C组(EC诱导组)及D组(BMSCs和诱导的EC联合培养组),通过干细胞形态、免疫荧光、细胞增殖、碱性磷酸酶活性及骨钙素含量从酶学、组织学及生化等不同方面观察诱导的EC对BM- SCs成骨活性及牛长情况的影响。结果细胞免疫荧光染色证实C组培养诱导的细胞为EC。倒置相差显微镜、HE染色均显示EC与BMSCs混合生长良好。MTF检测结果：各组细胞增殖差异无显著性意义(P＞0．05)。碱性磷酸酶活性和骨钙素含量检测结果：D组明显高于其它各组,差异有显著性意义(P＜0．05)。结论由BMSCs诱导的EC能够增强BMSCs的成骨活性,提高BMSCs的增殖能力。     

Perioperative and long-term morbidity and mortality after above-knee and below-knee amputations in diabetics and nondiabetics

We performed a retrospective review of a vascular surgery quality assurance database to evaluate the perioperative and long-term morbidity and mortality of above-knee amputations (AKA, n = 234) and below-knee amputations (BKA, n = 720) and to examine the effect of diabetes mellitus (DM) (181 of AKA and 606 of BKA patients). All patients in the database who had AKA or BKA from 1990 to May 2001 were included in the study. Perioperative 30-day cardiac morbidity and mortality and 3-yr and 10-yr mortality after AKA or BKA were assessed. The effect of DM on 30-day cardiac outcome was assessed by multivariate logistic regression and the effect on long-term survival was assessed by Cox regression analysis. The perioperative cardiac event rate (cardiac death or nonfatal myocardial infarction) was at least 6.8% after AKA and at most 3.6% after BKA. Median survival was significantly less after AKA (20 mo) than BKA (52 mo) (P < 0.001). DM was not a significant predictor of perioperative 30-day mortality (odds ratio, 0.76 [0.39-1.49]; P = 0.43) or 3-yr survival (Hazard ratio, 1.03 [0.86-1.24]; P = 0.72) but predicted 10-yr mortality (Hazard ratio, 1.34 [1.04-1.73]; P = 0.026). Significant predictors of the 30-day perioperative mortality were the site of amputation (odds ratio, 4.35 [2.56-7.14]; P < 0.001) and history of renal insufficiency (odds ratio, 2.15 [1.13-4.08]; P = 0.019). AKA should be triaged as a high-risk surgery while BKA is an intermediate-risk surgery. Long-term survival after AKA or BKA is poor, regardless of the presence of DM.     

Body composition assessment and methodology in nonathletic and athletic adolescent and adult males and females

The purpose of this review is to outline methodology for assessing body composition utilizing anthropometric and densitometric techniques. The objective of body composition assessment is to measure body fat and lean body mass. The quantity of these components varies due to growth, physical activity, dietary regimens, and aging. Anthropometric techniques incorporate selected skinfolds, circumferences, skeletal widths, or other variables to estimate body composition within k2.0-4.0%. These techniques are adequate for field testing of groups or individuals, but are population specific. Densitometry measures body volume irrespective of physique, sex, or age. This laboratory technique estimates body composition within 1.0-2.0%, is more difficult to administer, but is not population specific. Some limitation exists with any present technique due to biological variability and incomplete research of reference body composition in children, females, and the aged. J Orthop Sports Phys Ther 1984;5(6):336-347.     

Postoperative nausea and vomiting and opioid-induced nausea and vomiting: guidelines for prevention and treatment

Postoperative nausea and vomiting (PONV) causes patient discomfort, lowers patient satisfaction, and increases care requirements. Opioid-induced nausea and vomiting (OINV) may also occur if opioids are used to treat postoperative pain. These guidelines aim to provide recommendations for the prevention and treatment of both problems. A working group was established in accordance with the charter of the Sociedad Espa?ola de Anestesiología y Reanimación. The group undertook the critical appraisal of articles relevant to the management of PONV and OINV in adults and children early and late in the perioperative period. Discussions led to recommendations, summarized as follows: 1) Risk for PONV should be assessed in all patients undergoing surgery; 2 easy-to-use scales are useful for risk assessment: the Apfel scale for adults and the Eberhart scale for children. 2) Measures to reduce baseline risk should be used for adults at moderate or high risk and all children. 3) Pharmacologic prophylaxis with 1 drug is useful for patients at low risk (Apfel or Eberhart 1) who are to receive general anesthesia; patients with higher levels of risk should receive prophylaxis with 2 or more drugs and baseline risk should be reduced (multimodal approach). 4) Dexamethasone, droperidol, and ondansetron (or other setrons) have similar levels of efficacy; drug choice should be made based on individual patient factors. 5) The drug prescribed for treating PONV should preferably be different from the one used for prophylaxis; ondansetron is the most effective drug for treating PONV. 6) Risk for PONV should be assessed before discharge after outpatient surgery or on the ward for hospitalized patients; there is no evidence that late preventive strategies are effective. 7) The drug of choice for preventing OINV is droperidol.     

Similarities and differences between female and male sexual functions and dysfunctions

Men and women have 23 pairs of chromosomes. They share 22 of them. In physiologic conditions they differ systematically in only one pair, the sexual one. Females (normally) have what is called an “XX” on the 23rd pair of chromosomes, whereas males have an “XY” pair. The striking sexual differences –anatomic, functional, reproductive, psychological and sociocultural - between men and women depends on or derive from the difference in one critical chromosome out of 46, which contains on average 2% of all the genetic code. Biochemical, neuroendocrine, hormonal, vascular, nervous, and metabolic similarities that both sexes share, based on the common 45 chromosomes and related biologically determined similarities contributing to the secret sexual symmetry between genders, is reviewed. Furthermore the role of the genetically determined brain and somatic gender dymorphism, contributing to gender sexual differences is analyzed. Neuroplasticity and psychoplasticity are praised as basic mechanisms that bridge together and re-shape the individual biological and psychological world through the continuous interaction with the environment. Enhancement of sexual differences in behaviour, meaning of, and motivation to sex by cultural constructs, by religious and social dynamics, and the continuous interaction of each person with a usually role-polarized society during the whole life span will be finally acknowledged. To contribute to a better understanding of the shared biological sexual similarities between genders and their dialectic and continuous relation with biological and socioculturally related sexual differences is the ultimate goal of this introductory article and the following papers of the series.