A preliminary in vitro study on the fabrication and tissue engineering applications of a novel chitosan bilayer material as a scaffold of human neofetal dermal fibroblasts

The bilayer structure of chitosan film and sponge was designed as a scaffold of skin tissue engineering and a dermis substitute. It was processed successively via the formation of a dense chitosan film by casting method and a porous chitosan sponge by lyophilization. The dry thickness of the film layer was 19.6 microm and that of the sponge layer was controlled at 60-80 microm. Porogens such as sodium chloride, glucose, and sucrose were used to create large pores of the chitosan sponge layer. Human neofetal dermal fibroblasts were seeded in the chitosan sponge layer and cultured for 4 weeks. It was found that the cells could grow and proliferate well in an extended shape on the flat bottom of large pores with 15-100 microm width and in spherical form on the rough pore walls or at the edges of micropores less than 5 microm. Fibroblasts after the culture could bind tightly with the sponge layer via newly formed extracellular matrices to give a living cell-matrix-chitosan composite. The bilayer chitosan material remained stable in shape and size during the cell culture. The results suggested that the bilayer chitosan material would be an alternative of collagen materials which was obviously contracted during cell culture.     

兔骨髓基质细胞诱导成骨细胞复合同种异体生物衍生骨实验研究

目的探讨体外培养的骨髓基质细胞与自体来源的生物衍生骨复合后的生长特性，为进一步研究骨髓基质细胞作为种子细胞，以及探索一种良好的支架材料提供实验依据。方法分离纯化兔骨髓基质细胞并诱导成成骨细胞后与同种异体来源的生物衍生骨复合后体外培养，并在相差显微镜和扫描电镜下观察其生长情况。结果骨髓基质细胞在生物衍生骨上贴附生长、增殖良好。结论骨髓基质细胞可作为骨组织工程的理想种子细胞，与生物衍生骨复合后可作为骨组织工程的载体。     

全髋关节置换术均衡双下肢长度的探讨

目的探讨全髋关节置换术均衡双下肢长度的临床意义和手术设计。方法回顾分析2000-2004年获得随访的单侧全髋关节置换手术病例48例,采用骨水泥型假体12髋,混合型假体36髋,术前双下肢等长30例(含股骨颈骨折患者),10例患肢长度短缩1.0-2.0cm,6例短缩2.0-4.0cm,2例短缩4.0-6.0cm。通过术前肢体长度测量及X片测量制定手术方案,选择假体、预估颈长和截骨平面;术中正确磨削髋臼,寻找最佳旋转中心,结合透视及特殊试验修正截骨平面,调节假体颈长,达到均衡双下肢长度的目的。结果全部病例采用Harris评分,优30例,良12例,中4例,差2例。术前等长30例巾术后出现5例手术侧延长1.0-2.0cm,1例延长2.5cm。术前不等长的18例中,术后恢复等长10例,6例手术侧延长1.0-2.0cm,2例术前重度短缩的先天性髋关节发育不良病人术后仍然短缩2.0-3.0cm。术后肢体等长35例Harris评分平均92.3分,术后肢体不等长13例Harris评分平均88.6分。结论全髋关节置换术的手术设计。手术技巧以及假体设计等诸多因素影响下肢长度,术后肢体不等长降低了患者的满意度。应重视术前测量及术中综合评价方法的运用,重视软组织平衡技术和肢长调节,掌握全髋置换术中均衡下肢长度的手术技术,进一步改善手术效果。     

Anti-HBV effects of CpG oligodeoxynucleotide-activated peripheral blood mononuclear cells from patients with chronic hepatitis B

Unmethylated CpG dinucleotides in bacterial DNA or synthetic oligodeoxynucleotides containing immunostimulatory CpG motifs (CpG ODN) are known as a potent Th1-like immune enhancer in vertebrates. Chronic hepatitis B is the immunocompromising condition. We therefore investigated the effects of CpG ODN on cultured cells from chronic hepatitis B patients and healthy controls. The inhibitory effects of CpG ODN on hepatitis B virus (HBV) were also studied. The secretion of IFN-alpha by CpG ODN-activated peripheral blood mononuclear cells (PBMCs) from chronic hepatitis B patients and healthy controls was significantly increased when compared with PBMCs alone or GpC ODN-stimulated PBMCs. After activation with CpG ODN, the IFN-alpha secretion by chronically HBV-infected patient PBMCs is less than that by healthy control PBMCs. Treatment of HepG2 2.2.15 cells with culture supernatants of PBMCs activated by CpG ODN can significantly suppress the secretion of HBsAg, HBeAg and HBV DNA as compared with that of PBMCs without CpG ODN activation under the same conditions. No inhibitory effect on the replication of HBV was found for CpG ODN treatment alone. Our results indicated that CpG ODN could efficiently enhance the immune response of chronic hepatitis B patients. Moreover, the CpG ODN-activated PBMCs from chronic hepatitis B patients were able to significantly inhibit HBV replication in vitro, suggesting that CpG ODN may be a potential immunoregulator against HBV infection in the future.     

Rapid identification and susceptibility testing using the VITEK(R) 2 system using culture fluids from positive BacT/ALERT(R) blood cultures

BACKGROUND AND PURPOSE: In order to reduce the turnaround time for laboratory diagnosis of bacteremia, the efficacy of identification and antimicrobial susceptibility testing using samples taken directly from positive BacT/ALERT(R) standard aerobic and standard anaerobic blood culture bottles was evaluated. METHODS: 160 positive blood culture bottles were examined and incubated at 35 degrees C in 5% carbon dioxide for 4-24 h, and an aliquot of the culture fluid was Gram stained. Samples containing Gram-negative bacilli were inoculated on VITEK(R) 2 ID-GNB (identification-Gram-negative bacilli) and AST (antimicrobial susceptibility testing)-GN04 cards, and those containing Gram-positive cocci were inoculated on ID-GPC (identification-Gram-positive cocci) and AST-P526 cards. The same samples were also examined by the standard method, involving subculture from positive BacT/ALERT standard blood culture bottles. RESULTS: Eighty seven of 97 Gram-negative bacilli (89.7%) and 21 of 63 Gram-positive cocci (33.3%) were correctly identified to the species level. For antimicrobial susceptibility testing, the direct method had an overall error rate of 5.4% for Gram-negative bacilli, with 0.9% very major, 0.9% major, and 3.6% minor discrepancies compared to the standard method. The overall error rate in antimicrobial susceptibility testing for the 13 Staphylococcus spp. was 10.3%, with 6.0% very major, 2.6% major, and 1.7% minor discrepancies. CONCLUSION: These data suggest that VITEK 2 cards inoculated with samples taken directly from positive Bact/ALERT blood culture bottles would provide acceptable identification and antimicrobial susceptibility testing results for Gram-negative bacilli, but not for Gram-positive cocci. Compared to the standard method, the direct method would reduce turnaround time by at least 24 h.     

大鼠腘窝淋巴结内淋巴滤泡的生后发育

目的:研究大鼠淋巴结淋巴滤泡的生后发育.方法:采用常规组织学、免疫细胞化学及三维重建技术研究了大鼠腘窝淋巴结内淋巴滤泡的发生.结果:生后18天始,淋巴结浅层皮质内sIgM阳性B淋巴细胞聚集形成初级淋巴滤泡.生后3周ED-5阳性滤泡树突状细胞出现.随着鼠龄及体重的增长,初级淋巴滤泡不断扩大,每个淋巴结内淋巴滤泡数亦增多.生后13周滤泡数达高峰,平均每个淋巴结86个,13周以后体重缓慢增长,淋巴滤泡数逐渐下降.生后8周时,次级淋巴滤泡出现,ED-5阳性滤泡树突状细胞主要分布于亮区.结论:初级淋巴滤泡形成过程中B淋巴细胞聚集可能诱导局部网状细胞分化成滤泡树突状细胞;发育期间淋巴滤泡数随体重增长而增加可能与某些决定机体生长的因素作用有关.     

Mcm1 regulates donor preference controlled by the recombination enhancer in Saccharomyces mating-type switching

Switching of Saccharomyces mating type by replacement of sequences at the MAT locus involves a choice between two donors, HML and HMR. MATα cells inhibit recombination along the entire left arm of chromosome III, including HML, whereas MATa cells activate this same region. MATa-dependent activation of HML depends on a small, cis-acting DNA sequence designated the recombination enhancer (RE), located 17 kb centromere-proximal to HML. A comparison of RE sequences interchangeable between Saccharomyces cerevisiae and Saccharomyces carlsbergensis defines a minimum RE of 244 bp. RE activity is repressed in MATα cells by binding of the Matα2–Mcm1 corepressor to a site within the RE. Mutation of the two Matα2 binding sites removes most, but not all, of this repression, and RE chromatin structure in MATα cells becomes indistinguishable from that seen in MATa. Surprisingly, a 2-bp mutation in the Mcm1 binding site completely abolishes RE activity in MATa cells; moreover, RE chromatin structure in the MATa mutant becomes very similar to that seen in MATα cells with a normal RE, displaying highly ordered nucleosomes despite the absence of Matα2. Further, a mutation that alters the ability of Mcm1 to act with Matα2 in repressing a-specific genes also alters donor preference in either mating type. Thus, Mcm1 is critically responsible for the activation as well as the Matα2-Mcm1-mediated repression of RE activity.     

Skeletal alterations in hypophysectomized rats: I. A histomorphometric study on tibial cancellous bone

Background: Hypophysectomy (HX) results in a cessation of bone growth and a decrease in bone metabolism. The purpose of this study is to examine the effect of HX on the static and dynamic histomorphometry of cancellous bone in the secondary spongiosa of the proximal tibial metaphysis in rats. Methods: Female rats, at 2 or 3 months of age, were HX and sacrificed at 0, 5 days, 2 and 5 weeks after the surgery. Age-matched intact rats served as controls. Cancellous bone histomorphometry was performed on doublefluorescent labeled, 30-um-thick sections of the proximal tibia. Tartrateresistant acid phosphatase histomorphometry was performed at 5 days on HX and control rats to evaluate the resorption in the metaphyseal bone. Results: Although the intact rats gained in body weight, tibial length, tibial weight, and density after 5 weeks, these changes did not occur following HX. As compared to the basal group, HX resulted in a decrease in the density and dry weight of the metaphysis. The histomorphometric data showed that the cancellous bone volume and trabecular number of the secondary spongiosa were decreased and the separation was increased in the HX rats. The dynamic results showed that HX significantly decreased longitudinal growth rate and tissue-based bone formation and resorption. However, the bone surface-based eroded surface, labeled surface, the mineral apposition rate, and the bone formation rate did not differ between the intact and the HX rats at either the 2 or 5 weeks study. Five days after HX, the bone surface and tissue-based osteoclast surfaces were significantly lower in the HX than in the intact rats. Conclusions: Pituitary hormone deficiency results in cancellous bone loss. The bone loss is due primarily to the suppression of longitudinal growth-dependent bone gain and the inhibition of tissue-based bone turnover with a lower bone formation relative to bone resorption. The surfacebased bone turnover is not affected. © 1995 Wiley-Liss, Inc.     

Evaluation of the common conditions associated with eosinophilia.

In an eosinophilic population of 47 boys of the same age, a large proportion (92%) were helminth infested or atopic, or both, compared with 36% of 36 controls. The methods used to detect these conditions were not costly or elaborate, except for the radioallergosorbent test, which was used to measure concentrations of circulating IgE antibodies to atopic allergens. It is suggested that an economical approach to detect helminthiasis and atopy in cases of eosinophilia is adopted using the methods employed here, with skin prick tests replacing the radioallergosorbent test.