骨肉瘤成骨细胞表型与临床因素的相关性研究

目的采集手术切除未经培养的骨肉瘤标本,研究肿瘤中成骨细胞的分化及其与临床因素的关系。方法观察骨肉瘤中成骨细胞相关基因表达的频率、特异性,以及骨肉瘤成骨细胞表型与生物学行为的关系。应用RT-PCR方法研究48例骨肉瘤标本的碱性磷酸酶(ALP)、骨钙素(OC)和骨桥蛋白(ON)的mRNA转录。磷酸甘油酸激酶(PGK)被用于内参照。结果在大多数骨肉瘤标本中可以发现ALP或OC的mRNA转录(分别为93.75%,79.17%)。在所有的肉瘤检测中均可发现ON的mRNA转录。在骨肉瘤的成骨细胞亚型中可见到ALP和OC的高水平表达。肺转移瘤OC的mRNA表达弱于原发灶。肿瘤细胞有高水平OC表达的病例组有较高的生存率(P=0.045)。结论OC是目前最好的成骨细胞特异性标记物,将来可能用于检测骨肉瘤患者外周血中循环的肿瘤细胞。     

319例352眼眼外伤疗效观察

采用综合治疗，抗炎，散瞳，局部治疗与全身应用免疫抑制疗法，治疗３１９例３５２眼外伤获得了满意的疗效，眼球挫伤１０４例，眼球穿孔伤９３例，化学烧伤７１例，眼炸伤４６例。眼热烧伤５例，治疗包括及时清创胶粘或缝合，清除前房出血合理用皮质类固醇和免疫抑制剂等。眼球挫伤治愈率９７．１２％，穿透伤９３．１８％，热烧伤６６．６７％。３１９例眼外伤治愈率９４．６７％好转５．３３％，并对各种治疗方法进行了讨论。     

Measurement of blood loss at childbirth and postpartum.

OBJECTIVE: To accurately measure blood loss during childbirth in a developing country. METHOD: The alkaline hematin technique was used to quantify blood lost during delivery and 24 h postpartum in 158 women in Pemba Island, Zanzibar. RESULT: Women were found to lose less blood during childbirth and 24 h postpartum than previously reported. Compared with laboratory values, nurse-midwives approximated blood loss accurately (mean difference, i.e., mean underestimation by nurse-midwives, 4.90 mL); however, their imprecision was greater for higher laboratory values. CONCLUSION: This study may prompt further investigation, as no comparable data exist for developing countries where maternal mortality is high and severe anemia prevalent.     

大白鼠皮肤Ⅱ度、Ⅲ度碱烧伤模型复制

目的 根据大白鼠皮肤碱烧伤创面早期病理组织学观察复制Ⅱ、Ⅲ度碱烧伤模型,研究碱烧伤的临床治疗.方法 Wistar纯种健康大白鼠20只,以不同浓度2mol/l、2.5mol/l、5mol/lNaOH,作用时间分别为30秒、45秒、60秒、75秒,涂抹于脱毛后的表皮.结果 2mol/L(60s)、2.5mol/L(45s),5mol/L(30s)即可直接造成Ⅱ度～深Ⅱ度烧伤;2mol/L(75s)、2.5mol/L(60s)、5 mol/L(45s)即可造成Ⅲ度烧伤,其烧伤深度与NaOH溶液浓度和作用时间呈正比.结论 Ⅱ度、Ⅲ度碱烧伤均为渐进性烧伤,与文献中介绍的潜拙样损伤似乎不同.另外,不同浓度NaOH溶液在相同时间对皮肤组织的损伤及同一浓度NaOH溶液在不同时间对大白鼠皮肤的损伤病理学变化均有差异.     

生长板软骨细胞促进骨髓基质干细胞血管内皮生长因子的表达

目的观察骨髓基质干细胞(BMSCs)在生长板软骨细胞旁分泌作用下血管内皮生长因子(VEGF)的表达规律及其与成骨分化的相关性。方法大鼠BMSCs与生长板软骨细胞进行间接共培养,培养终末期做细胞化学染色,定量测定碱性磷酸酶(ALP)活性,用RT-PCR方法半定量检测VEGFmRNA的表达。结果生长板软骨细胞持续高表达VEGF。BMSCs随共培养时间的延长,ALP活性升高,BMSCs的VEGF的表达也逐渐增强。培养液加入两种分泌型VEGF中和抗体后,VEGF表达趋势不变,ALP活性仍为升高趋势,也不影响培养终末期钙化结节的形成。培养终末期BMSCs的CD31和CD34均阴性。结论BMSCs成骨分化过程中VEGF的表达符合成骨细胞分化基因的表达规律,与成骨细胞特征性基因的表达趋势一致,体外条件共培养条件下,中和VEGF后并不能阻碍BMSCs的成骨分化。     

新型生物陶瓷对体外培养成骨细胞影响的试验研究

目的 :研究新型复合生物陶瓷 (ZrO2 -CBC)对体外培养的胎鼠成骨细胞增殖分化的影响 ,探讨ZrO2 -CBC的细胞相容性 .方法 :将ZrO2 -CBC与胎鼠成骨细胞体外共同培养 15d ,通过细胞计数、ALP活性检测和流式细胞仪法评价ZrO2 -CBC对成骨细胞增殖分化的影响 .结果 :与空白对照组相比 ,实验早期ZrO2 -CBC对成骨细胞增殖分化无抑制作用 (P >0 0 5 ) ,后期轻度抑制成骨细胞的增殖 ,并抑制成骨细胞ALP的分泌 (P <0 0 5 ) .结论 :ZrO2 -CBC是一种有前途的牙科种植体材料     

Diagnosis of gallbladder perforation in acute acalculous cholecystitis in critically ill patients

In the presence of ascites ultrasound is not appropriate to distinguish between gallbladder perforation and acute acalculous cholecystitis. However, the correct and early diagnosis of gallbladder perforation is important for the treatment and prognosis. We report 4 critically ill patients with ascites. All patients had evidence of gallbladder perforation by ultrasound and underwent cholecystectomy: 2 patients had gallbladder perforation, but 2 had acalculous cholecystitis without perforation. markedly elevated serum alkaline phosphatase was the only discriminating finding indicating gallbladder perforation.     

Stable Transfection of Nonosteogenic Cell Lines with Tissue Nonspecific Alkaline Phosphatase Enhances Mineral Deposition Both in the Presence and Absence of β-Glycerophosphate: Possible Role for Alkaline Phosphatase in Pathological Mineralization

It is documented that alkaline phosphatase (AP) plays an important role in bone mineralization. Considering that TN-AP is expressed in periodontal ligament fibroblasts, renal epithelial cells, and vascular endothelial cells, and that TN-AP is both a calcium-/phosphate-binding protein and a phosphohydrolytic enzyme, we hypothesize that membrane-bound AP also plays an important role in the initiation of physiological and pathological mineralizations in tissues other than bone and cartilage. To test this hypothesis, nonosteoblast cell lines, including a fibroblast line, a renal epithelial line, and a capillary endothelial line, were stably transfected to express high levels of rat bone AP on their cell surfaces. These rat bone AP-expressing cells were then cultured on filter membranes in the presence or absence of β-glycerol phosphate. von Kossa staining for calcium phosphate and transmission electron microscopy with electron diffraction analysis for minerals were employed to investigate the effect of membrane AP on extracellular calcium phosphate mineralization. Our results indicated that AP expression on these nonosteoblast-like cell surfaces have induced extracellular hydroxyapatite (HAP) mineralization. Our findings support the concept that membrane-bound AP contributes to extracellular apatitic mineralization by mechanisms that do not necessarily involve its hydrolase activity. They also suggest that AP might be important for the initiation of pathological mineralization in nonosteogenic tissues. Received: 11 January 1996 / Accepted: 31 October 1996     

猪肾碱性磷酸酶的提纯及性质研究

目的：从猪肾中提取碱性磷酸酶(ALP)，并对其能否作为参考品进行研究。方法：从猪肾皮质中分离刷状缘微绒毛膜，用正丁醇溶解膜结合的酶，经硫酸铵沉淀，DEAE-Sephacel、ConA-Sepharose、Sephadex G-200及抗γ-谷酰基转移酶(GGT)免疫亲和层析提纯ALP。结果：提纯的ALP比活达402kU／g蛋白，几乎不含杂酶(未检出ALT、AST、ChE、AMY、CK、LDH，GGT仅含2U／780U ALP)。其对底物4-硝基磷酸酚(4-NPP)的Km值为1．35mmol／L，最适pH值为10．40。结论：提纯的猪肾ALP与人血清中的ALP酶动力学特性非常相似，为进一步研制ALP参考品奠定了基础。     

Pulsed electromagnetic fields alter phenotypic expression in chondroblasts in tissue culture

We hypothesize that pulsed electromagnetic fields (PEMF) alter phenotypic expression of chondroblasts by promoting the production of alkaline phosphatase (AP) and altering the structure of proteoglycans. Chondroblasts from the hypertrophic zone of tibial epiphyses (HC), sternum (SC), and skin fibroblasts (F) were cultured from 16 day chick embryos. Cultures were randomly designated control (C) or experimental (E). E received PEMF for 24 h in a 6 h on, 6 h rest sequence. The controls were in the same incubator shielded by Mu metal. Assays for AP activity were performed and normalized to protein content. Proteoglycan synthesis assay involved labeling with 35S fractionating in a 5% to 20% surcrose gradient determining total protein and chondroitin sulfate content. PEMF showed no change of AP activity on F. A high AP basal activity was found in HC, but was not increased above the control. PEMF increased AP in the SC samples (E/C ratio). The sucrose gradient data showed a shift in peaks for SC only altering the ratio of carbohydrate to protein for the SC. Analysis of carbohydrate and protein indicated that the effect was decreased synthesis or degradation of protein. We conclude that PEMF alters the phenotypic expression of sternal chondroblasts in our in vitro system.