Dynamic changes of mechanically activated channels and K+ channels at injury site of peripheral nerve in rat

Ectopic ion channels developed locally at the injury site after nerve damage by light ligation around common sciatic nerve of the rats. Different channel types have different processes of formation, accumulation and degeneration. During the first three days after injury, mechanically activated channels that are modulated by Ca++ channel activities first appeared. As the nerve fibers begin to be excited by TEA, a blocker of K+ channels, suggesting that the accumulation of K+ channels, the responsibility of mechanically activated channels was declining. Onset of K+ channels was from the 3rd postoperative day and lasted up to the fiftieth day. This time course of K+ channel development was closely related to allodynia and hyperalgesia of neuropathic animal behaviour. The results suggest that chronic contraction injury induces a dynamic change in the ectopic mechanically activated channels and K+ channels at the injury site of nerve and there is an interchange in the development time courses of the mechanic     

1α,25-dihydroxy-24-oxo-16-ene vitamin D3, a metabolite of a synthetic vitamin D3 analog, 1α,25-dihydroxy-16-ene vitamin D3, is equipotent to its parent in modulating growth and differentiation of human leukemic cells

1α,25(OH)₂-16-ene-D₃, a synthetic analog of the steroid hormone, 1α,25(OH)₂D₃, has great potential to become a drug in the treatment of leukemia and other proliferative disorders, because of its minimal in vivo calcemic activity associated with a potent inhibitory effect on cell growth. However, at present, the mechanisms through which 1α,25(OH)₂-16-ene-D₃ expresses its biological activities are still not completely understood. Our previous in vitro study in a perfused rat kidney indicated for the first time that 1α,25(OH)₂-16-ene-D₃ and 1α,25(OH)₂D₃ are metabolized differently. 1α,25(OH)₂-24-oxo-16-ene-D₃, an intermediary metabolite of 1α,25(OH)₂-16-ene-D₃ formed through the C-24 oxidation pathway, accumulated significantly in the perfusate when compared to 1α,25(OH)₂-24-oxo-D₃, the corresponding intermediary metabolite of 1α,25(OH)₂D₃. In a subsequent in vivo study, we also reported that 1α,25(OH)₂-24-oxo-16-ene-D₃ exerted immunosuppressive activity equal to its parent, without causing significant hypercalcemia. In order to establish further the critical role of 1α,25(OH)₂-24-oxo-16-ene-D₃, in generating some of the key biological activities ascribed to its parent, we performed the present in vitro study using a human myeloid leukemic cell line (RWLeu-4) as a model. Comparative target tissue metabolism studies indicated that 1α,25(OH)₂-16-ene-D₃ and 1α,25(OH)₂D₃ are metabolized differently in RWLeu-4 cells, and the differences were similar to the ones we previously observed in the rat kidney. The significant finding was the accumulation of 1α,25(OH)₂-24-oxo-16-ene-D₃ in RWLeu-4 cells because of its resistance to further metabolism. Biological activity studies indicated that both 1α,25(OH)₂-16-ene-D₃ and its 24-oxo metabolite produced growth inhibition and promoted differentiation of RWLeu-4 cells to the same extent, and these activities were several fold higher than those exerted by 1α,25(OH)₂D₃. In addition, the genomic action of each vitamin D compound was assessed in a rat osteosarcoma cell line (ROS 17/2.8) by measuring its ability to transactivate a gene construct containing the vitamin D response element of the osteocalcin gene linked to the growth hormone reporter gene. In these studies, both 1α,25(OH)₂-16-ene-D₃ and its 24-oxo metabolite exerted similar but potent transactivation activity which was several fold greater than that exerted by 1α,25(OH)₂D₃ itself. In summary, our results indicate that the production and slow clearance of the bioactive intermediary metabolite, 1α,25(OH)₂-24-oxo-16-ene-D₃, in RWLeu-4 cells contributes significantly to the final expression of the enhanced biological activities ascribed to its parent analog, 1α,25(OH)₂-16-ene-D₃.     

微生物产生的木聚糖酶的功能和应用

术聚糖是一种异质多糖,主要由木糖和阿拉伯糖组成。微生物产生的木聚糖酶来源广泛,能将木聚糖水解为木寡糖和D-木糖。该酶具有极大的应用价值,如可用于纸浆的漂白以减少环境污染,也可将造纸工业及农业废料中的木聚糖转化为D-木糖。     

一种模拟高原低氧复合冷冻的简易动物实验舱

以氮稀释法使舱内氧含量维持实验模拟高度,以普通冰箱制冷系统维持舱内恒温,以多功能低温浴槽作为致冻装置。该舱可于２０ｍｉｎ内升至模拟６０００ｍ高度,每次可供１６只大鼠作高原冻伤实验。实验期间高度恒定于６０００±２００ｍ,舱温２１±１℃。相对湿度５５％±１５％,冷冻液－２５．０±０．５℃。该舱结构简单、使用方便,可在实验室中模拟高原条件进行冷冻实验研究。由于不使用人体减压舱,不仅避免了减压缺氧对实验人员的不利影响,也减少了实验消耗。     

一类竞争扩散方程组在Neumann边条件下解的渐近行为

本文主要采用比较方法讨论了一类竞争扩散方程组在Neumann边条件下解的渐近行为,得到了解的稳定性区域．     

肝炎平对D－氨基半乳精所致肝损害保护作用的酶组织化学研究

本实验采用Ｄ－氨基半乳糖（Ｄ－ＧａｌＮ）诱导的大鼠急性肝损伤模型,观察大鼠肝脏组织化学的变化,探讨肝炎平对急性肝损伤的保护作用。实验分为四组,即正常对照组、模型组、肝炎平及肝得健保护组。结果表明：肝炎平对肝细胞膜系统有一定的保护作用。肝炎平组和肝得健组ＳＤＨ、ＣＣＯ及ＣｈＥ活性明显高于模型组,且与正常对照组相近。本实验模型组ＡＣＰ的活性明显高于正常组,而肝炎平组ＡＣＰ的活性明显低于模型组,与正常对照组无显著性差异。提示：肝炎平可显著改善因Ｄ－氨基半乳糖所致肝损害的作用。且其对肝细胞的保护作用与肝得健一致。     

HPI在肝癌组织中的表达及其对肝癌细胞特异性抑增殖效应研究

肝细胞增殖抑制因子（Ｈｅｐａｔｉｃｐｒｏｌｉｆｅｒａｔｉｏｎｉｎｈｉｂｉｔｏｒ,ＨＰＩ）粗制品、半纯品和纯品对体外培养的人肝癌细胞具有显著抑增殖作用,随样品纯度提高抑制活性逐渐增强。纯品（浓度５μｇ／ｍｌ）的抑制率达７７．７１％。正常成年大鼠肝细胞呈ＨＰＩ阳性表达。在ＤＥＮ诱发大鼠肝细胞癌的发生发展过程中,转化的癌前期细胞和肝癌细胞呈ＨＰＩ阴性表达。表明肝细胞ＨＰＩ的表达能力在其癌变过程中消失,从而失去了自身的抑癌作用。     

RT－cDNA克隆－PCR法获取犬瘟热病毒融合蛋白基因（F）

纯化鸡胚成纤维细胞培养的犬瘟热病毒（ＣａｎｉｎｅＤｉｓｔｅｍｐｅｒＶｉｒｕｓ,ＣＤＶ）,获得病毒基因组ＲＮＡ后,反转录合成双链病毒Ｆ基因ｃＤＮＡ。将此双链ｃＤＮＡ平端插入ＰＵＣ１９质粒ＳａｍⅠ位点构建重组质粒,进行ｃＤＮＡ克隆。以重组克隆质粒为模板ＰＣＲ扩增,获得ＣＤＶ全长Ｆ基因。将此Ｆ基因插入表达载体ＰＢＶ２２０,在大肠杆菌中表达,通过对表达产物的最终鉴定,可确认所获片段为ＣＤＶ全长Ｆ基因．     

固定化酵母细胞生产1,6-二磷酸果糖研究

本文研究了固定化酵母细胞制备果糖1,6二磷酸(FDP)的方法及其生产。用卡拉胶包埋方法固定化酿酒酵母(Sacchromyces cerevisae),对含葡萄糖1.0M,磷酸盐0.8M的糖磷液,pH6.5,在37℃下进行磷酸化反应。反复分批转化20天以上,可达到平均产FDPH_427.58mg/ml,最高为59.94mg/ml。用100ml固定化细胞生物反应器连续运转309h,稀释速率D=0.097h~(-1),平均产FDPH_4 21.51mg/ml。20L反应器连续运转,生产能力达到1.7g/h.L。用层析方法制备FDPNa_3结晶粉,提取收率为72.08％,制备质量达到或超过了国内外同类产品的质量要求。     

DNA fingerprinting of human cell lines using PCR amplification of fragment length polymorphisms

Summary Methods for monitoring cell line identification and authentication include species-specific immunofluorescence, isoenzyme phenotyping, chromosome analysis, and DNA fingerprinting. Most previous studies of DNA fingerprinting of cell lines have used restriction fragment length polymorphism analysis. In this study, we examined the utility of an alternative and simpler method of cell line DNA fingerprinting—polymerase chain reaction (PCR) amplification of fragment length polymorphisms. Fourteen human cell lines previously found by other methods to be either related or disparate were subjected to DNA fingerprinting by PCR amplification of selected fragment length polymorphism loci. Cell identification patterns by this method were concordant with those obtained by isoenzyme phenotyping and restriction fragment length polymorphism-DNA fingerprinting, and were reproducible within and between assays on different DNA extracts of the same cell line. High precision was achieved with electrophoretic separation of amplified DNA products on high resolution agarose or polyacrylamide gels, and with fragment length polymorphism (FLP) loci-specific “allelic ladders” to identify individual FLP alleles. Determination of the composite fingerprint of a cell line at six appropriately chosen fragment length polymorphism loci should achieve a minimum discrimination power of 0.999. The ability of PCR-based fragment length polymorphism DNA fingerprinting to precisely and accurately identify the alleles of different human cell lines at multiple polymorphic fragment length polymorphism loci demonstrates the feasibility of developing a cell line DNA fingerprint reference database as a powerful additional tool for future cell line identification and authentication.