Novel method for the quantitative assessment of cell migration: a study on the motility of rabbit anterior cruciate (ACL) and medial collateral ligament (MCL) cells

A novel method of quantitating cell migration has been proposed for the potential utilization of tissue engineered scaffolds. Applying Alt's conservation law to describe the motion of first passage ACL and MCL cells, we have developed a quantitative method to assess innate differences in the motility of cells from these two ligamentous tissues. In this study, first passage ACL and MCL cells were cultured from four mature New Zealand white rabbits. One side of the cell monolayer was scraped completely away to create a wound model. The cell moved into the cell-free area, and cell density profiles were analyzed at 6 h and 12 h. Values of the random motility coefficient (mu) were then estimated by curve fitting the 6 h and 12 h data to a mathematical model, derived from the conservation law of cell flux. During 6 h of incubation in medium supplemented with 1% FBS, MCL cells (mu(MCL) = 4.63 +/- 0.65 X 10(-6) mm(2)/sec) were significantly (p < 0.05) more mobile than ACL cells (mu(ACL) = 2.51 +/- 0.31 X 10(-6) mm(2)/sec). At 12 h, the MCL cells also appeared to move faster (mu(ACL) = 4.39 +/- 0.63 X 10(-6) mm(2)/sec, mu(MCL) = 6.59 +/- 1.47 X 10(-6) mm(2)/sec), but the difference was not statistically significant (p = 0.18). Exposure of the cells to growth factors PDGF-BB or bFGF for 6 h had no significant effect on the migration of the ACL and MCL cells. However, exposure of the ACL cells (p < 0.05) and the MCL cells (p = 0.19) to 1 ng/mL of PDGFBB for 12 h enhanced their migration. Incubation with a high concentration (100 ng/mL) of PDGF-BB or bFGF at concentrations tested (1 or 100 ng/mL) for 12 h, produced little or no migratory stimulation on these ligament cells. Our findings support the previous qualitative observations made by numerous investigators. The novel methodology developed in this study may provide a basis for tissue engineering, and the results may be applied to tissue reconstruction techniques of the knee ligaments.     

Human hepatic lipase mutations and polymorphisms.

Human hepatic lipase (HL) is a 477 residue glycoprotein that hydrolyzes triglycerides from plasma lipoproteins. Familial HL deficiency is a rare recessive disorder that is characterized by premature atherosclerosis and abnormal circulating lipoproteins. While studying the HL gene from the world's index family with HL deficiency, we identified four coding sequence variants of HL, one in each of exons 4, 5, 6, and 8. In this report we present the genetic basis for two new HL gene variants, one in each of exons 3 and 5. All six HL DNA variants are single base pair changes. Two variants (at codons 133 and 202) are diallelic DNA polymorphisms that are silent at the amino acid level. One variant (V73M) is an allele that defines an uncommon HL isoprotein. One variant (N193S) has two alleles of approximately equal frequency in the population that specify two common HL isoproteins. Two variants (S267F and T383M) are rare mutations found to date only in HL deficient subjects and their relatives. Of the six HL variants described to date, only S267F and T383M are associated with hyperlipidemia.     

腕关节关节囊内韧带的解剖观察及其创伤学意义

本文描绘了腕关节关节囊内韧带的解剖,其中首次描述了中腕关节的韧带连结,即大、小多角骨,头状骨至舟状骨的韧带和钩骨至三角骨的韧带.本文还论述了与这些韧带连结方式密切相关的腕骨生理运动特点及维持腕关节稳定性的因素.文章还讨论了与腕关节关节囊内韧带有关的腕部创伤学特点.     

An animal model of SARS produced by infection of Macaca mulatta with SARS coronavirus

A new SARS animal model was established by inoculating SARS coronavirus (SARS-CoV) into rhesus macaques (Macaca mulatta) through the nasal cavity. Pathological pulmonary changes were successively detected on days 5-60 after virus inoculation. All eight animals showed a transient fever 2-3 days after inoculation. Immunological, molecular biological, and pathological studies support the establishment of this SARS animal model. Firstly, SARS-CoV-specific IgGs were detected in the sera of macaques from 11 to 60 days after inoculation. Secondly, SARS-CoV RNA could be detected in pharyngeal swab samples using nested RT-PCR in all infected animals from 5 days after virus inoculation. Finally, histopathological changes of interstitial pneumonia were found in the lungs during the 60 days after viral inoculation: these changes were less marked at later time points, indicating that an active healing process together with resolution of an acute inflammatory response was taking place in these animals. This animal model should provide insight into the mechanisms of SARS-CoV-related pulmonary disease and greatly facilitate the development of vaccines and therapeutics against SARS.     

Fatal avian influenza A (H5N1) in a child presenting with diarrhea followed by coma

In southern Vietnam, a four-year-old boy presented with severe diarrhea, followed by seizures, coma, and death. The cerebrospinal fluid contained 1 white cell per cubic millimeter, normal glucose levels, and increased levels of protein (0.81 g per liter). The diagnosis of avian influenza A (H5N1) was established by isolation of the virus from cerebrospinal fluid, fecal, throat, and serum specimens. The patient's nine-year-old sister had died from a similar syndrome two weeks earlier. In both siblings, the clinical diagnosis was acute encephalitis. Neither patient had respiratory symptoms at presentation. These cases suggest that the spectrum of influenza H5N1 is wider than previously thought.     

EMS患者治疗前后血清SIL-2R、TNF-α和CA199检测的临床意义

目的:探讨子宫内膜异位症患者治疗前后血清SIL-2R、TNF-α和CA199水平的变化及意义.方法:应用放免法测定40例子宫内膜异位症患者血清TNF-α和CA199含量、酶联法测定SIL-2R含量,并与35名正常健康人作比较.结果:在治疗前,血清SIL-2R、TNF-α和CA199水平非常显著地高于正常人组(P＜0.01),经治疗1个月后,与正常人组比较仍有显著差异(P＜0.05).结论:检测子宫内膜异位症患者血清SIL-2R、TNF-α和CA199水平的变化对其病情和预后判断均有重要的临床意义.     

Oxygen-dependent up-regulation of mucoid exopolysaccharide (alginate) production in Pseudomonas aeruginosa.

We previously showed substantial differences in Pseudomonas aeruginosa exopolysaccharide production in vitro at oxygen tensions reflective of the right versus left cardiac circuits in vivo (40 versus 80 mm Hg, respectively; A. S. Bayer, T. O'Brien, D. C. Norman, and C. C. Nast, J. Antimicrob. Chemother. 23:21-35, 1989). However, those studies did not specifically confirm this exopolysaccharide to be the characteristic P. aeruginosa mucoid alginate seen in patients with cystic fibrosis. With a murine monoclonal antibody prepared against P. aeruginosa alginate, strongly positive immunofluorescence (IF) staining of a nonmucoid P. aeruginosa strain (PA-96) was seen after its exposure in vitro to oxygen tensions (pO2) of approximately 80 mm Hg; the intensity of the IF staining under these conditions was similar to that observed with a phenotypically mucoid P. aeruginosa strain (C1712M) from a cystic fibrosis patient. In contrast, the same nonmucoid strain (PA-96), after exposure to pO2 of approximately 40 mm Hg, showed little IF staining for alginate. Following enzyme treatment with alginase, PA-96 cells previously exposed to the higher pO2 and exhibiting enhanced alginate production, as determined by IF staining, now showed no IF staining. Moreover, treatment of the oxygen-up-regulated PA-96 cells with alginase released amounts of unsaturated alginate breakdown products (uronic acids) quantitatively similar to those released by typically mucoid strains treated with the same enzyme. These data indicated that the P. aeruginosa exopolysaccharide in our studies was, indeed, mucoid alginate and that variations in oxygen tensions represent one of the trigger mechanisms for the up-regulation of mucoid exopolysaccharide production.     

一种新型的脱细胞组织工程血管支架的构建和评价

本研究的目的是制备一种免疫原性较小、生物相容性较好、力学性能优良的组织工程血管支架。新鲜获得的犬主动脉,置于三蒸水中4℃过夜，使血管细胞由于渗透压差较大而破裂；随后经过多聚环氧化合物家族的乙二醇缩水甘油醚（EX-810）的作用。进一步促进细胞破裂的同时。对血管支架的纤维结构起交联作用；最后应用物理超声的方法清除支架内的细胞碎片残留。用这种方法处理的犬主动脉内几乎没有可见的核染，基本消除了血管支架的免疫原性。同新鲜的血管相比较。这种组织工程血管支架的各种力学指标与新鲜的犬主动脉没有显著差异。说明处理后的支架仍然保持新鲜血管的力学特征。同时它还表现出极低的细胞毒性。分别在支架上种植内皮细胞和平滑肌细胞，扫描电镜检测结果表明，两种细胞在支架上生长良好，且局部已经融合成片。 相容性     

维拉帕米对失血性休克犬心脏的保护作用及机制

杂种犬１１只，由股动脉放血使平均动脉压（ＭＡＰ）维持在６．０ｋＰａ９０ｍｉｎ后回输全部血液，继续观察１５０ｍｉｎ。在休克３０ｍｉｎ后，对照组和维拉帕米处理组（Ｖｅｒ组）分别静脉滴注生理盐水和Ｖｅｒ溶液［（１０μｇ／ｋｇ·ｍｉｎ）１５ｍｉｎ］，液体总量为３ｍｌ／ｋｇ。对照组犬在失血后心率（ＨＲ）加快，左室ｄｐ／ｄｔｍａｘ显著降低，回输血液后ＭＡＰ虽有回升，但仍低于基础值，左室ｄｐ／ｄｔｍａｘ无明显改善；Ｖｅｒ使休克犬ＨＲ减慢，血液回输后ＭＡＰ缓慢恢复至基础值，左室ｄｐ／ｄｔｍａｘ显著高于对照组；电镜观察可见对照组心肌肌原纤维和线粒体等有明显的损伤，而Ｖｅｒ组心肌超微结构基本正常。结果还显示：Ｖｅｒ组犬心肌中丙二醛含量、黄嘌呤氧化酶活性低于对照组，而超氧化物歧化酶活性高于对照组。结果表明：Ｖｅｒ对失血性休克犬的心脏具有保护作用，其机制可能与其阻滞膜Ｃａ２＋内流、抑制脂质过氧化有关。     

脑室注射促甲状腺素释放激素对电针镇痛及脑内亮氨酸脑啡肽含量的影响

实验用大鼠76只,用辐射热-甩尾法测痛,以电针后痛阈变化的百分率评价镇痛效应;用放射免疫分析法测定脑内L-Enk含量。发现脑室注入三肽-促甲状腺素释放激素(TRH)后,能显著降低电针镇痛效果;提高丘脑内L-Enk含量,但痛阈与脑内L-Enk含量的变化无相关关系。本文证明,TRH在电针镇痛过程中是对抗作用。已知针刺镇痛与脑内多种神经质和肽类有关,在不利于针刺镇痛的因素中,TRH是新添的“一员”。