重组人粒细胞集落刺激因子缓释微球的研究

目的：研究固体／油／水法制备重组人粒细胞集落刺激因子缓释微球,为开发其缓释剂型进行初步研究。方法：以聚乳酸．聚羟乙酸共聚物（PLGA）为载体材料：用固体／油／水法和水／油／水法制备载rhG-CSF缓释微球;考察粒径大小、外观、包封率等理化性质;用MieroBCA法考察微球的体外释药特性及影响因素;用SEC-HPLC和MTT比色法初步评价了微球制备工艺过程对rhG-CSF稳定性的影响。结果：两种方法制得的微球形态圆整、分散性良好,包封率均超过80％。固／油／水法制得的微球体外释放在2周内可超过90％,而水／油／水法制得的微球在相同的时间内仅释放30％。对于固／油／水法制备过程,SEC-HPLC法测定蛋白无明显聚集体出现,MTT法测定蛋白活性无明显损失。结论：实验证明了固／油／水法制备的PLGA微球可以实现2周以上的体外缓释。     

Atorvastatin suppresses oxidized LDL-induced dendritic cell-like differentiation of RAW264.7 cells regulated by the p38 MAPK pathway

The molecular signaling events leading to protection from oxidative stress-induced apoptosis upon contact inhibition have not been fully investigated. Previous research has indicated a role for mitogen-activated protein kinases (MAPKs) in the regulation of contact inhibition, and these proteins have also been associated with cell cycle regulation and stress-induced apoptosis. The potential role of the MAPK JNK-1 in the stress-response of actively proliferating and contact-inhibited cells was investigated. Actively proliferating normal fibroblasts (BJ) and fibrosarcoma cells (HT-1080) were stressed with H2O2, and levels of activated JNK-1 and cleaved PARP were ascertained. Similarly, these results were compared with levels of activated JNK-1 and cleaved PARP detected in H2O2-stressed confluent fibrosarcoma or contact-inhibited fibroblast cells. Contact-inhibited fibroblasts were protected from apoptosis in comparison to subconfluent fibroblasts, concurrent with decreased JNK-1 activation. Increased culture density of fibrosarcoma cells was not protective against apoptosis, and these cells did not demonstrate density-dependent alterations in the JNK-1 stress response. This decreased activation of JNK-1 in stressed, contact-inhibited cells did not appear to be dependent upon increased expression of MKP-1; however, over-expression of MKP-1 was sufficient to result in a slight decrease in H2O2-stimulated PARP cleavage. Increasing the antioxidant capacity of fibroblasts through NAC-treatment not only lessened H2O2-stimulated JNK-1 activation, but also did not influence the expression of MKP-1. Taken together, these results suggest that regulation of negative regulation of JNK-1 upon contact inhibition is protective against apoptosis, and that this regulation is independent of MKP-1.     

Carbachol increases Na+-HCO3- cotransport activity in murine colonic crypts in a M3-, Ca2+/calmodulin-, and PKC-dependent manner

The Na(+)-HCO(3)(-) cotransporter (NBC) mediates HCO(3)(-) import into the colonocyte via its pNBC1 isoform. Whereas renal kNBC1 is inhibited by increased cAMP levels, pNBC1 is stimulated. Cholinergic stimulation activates renal NBC, but the effect on intestinal NBC is unknown. Therefore, crypts were isolated from the murine proximal colon by Ca(2+) chelation and loaded with the pH-sensitive dye 2',7'-bis-carboxyethyl-5,6-carboxyfluorescein. Na(+)-HCO(3)(-) cotransport activity was calculated from the dimethylamiloride-insensitive (500 microM) intracellular pH recovery from an acid load in the presence of CO(2)-HCO(3)(-) and the intracellular buffering capacity. Carbachol strongly increased Na(+)-HCO(3)(-) cotransport activity compared with control rates. Ca(2+) chelation with BAPTA-AM, blockade of the M(3) subtype of muscarinergic receptors with 4-diphenylacetoxy-N-methylpiperidine methiodide, and inhibition of Ca(2+)/calmodulin kinase II with KN-62 all caused significant inhibition of the carbachol-induced NBC activity increase. Furthermore, PKC inhibition with G?-6976 and G?-6850 significantly reduced the carbachol effect, which may be related to the unique NH(2)-terminal consensus site for PKC-dependent phosphorylation of pNBC1. We conclude that NBC in the murine colon is thus activated by carbachol, consistent with its presumed function as an anion uptake pathway during intestinal anion secretion, but that the signal transductions pathways are distinct from those involved in the cholinergic activation of renal NBC1.     

Potent angiogenic inhibition effects of deacetylated chitohexaose separated from chitooligosaccharides and its mechanism of action in vitro

This study was performed to demonstrate the effects of deacetylated chitohexaose (hexamer) separated from a chitooligosaccharide (COS) mixture on tumor angiogenesis and its mechanism of action. Five fractions from dimer to hexamer were separated by a linear gradient solution of HCl on a cation-exchange resin. Then HCl was removed from the fractions by a charcoal column. The purity of the five fractions was analyzed by HPLC and the molecular masses were analyzed by MALDI-TOFMS. The hexamer expressed an inhibitory influence on CAM angiogenesis in a dose-dependent manner at concentrations of 6.25-50 μg/egg. On further investigation, we found that the hexamer had no toxic effect on normal ECV304 cells, but could inhibit the proliferation and migration of tumor-induced ECV304 cells in a dose-dependent manner. The mechanism was demonstrated through the detection of mRNA expression of VEGF, MMP-9, TIMP-1, TIMP-2, and uPA by RT-PCR, which showed that the hexamer down-regulated the VEGF and uPA mRNA expressions in ECV304 cells, but up-regulated the TIMP-1 mRNA expression.     

Prevalence of Age-Related Cataract in High-Selenium Areas of China

The objective of this study was to determine the prevalence of age-related cataract in high-selenium areas of China. This is a cross-sectional study of 1,522 persons aged 50 years and more who were selected as a representative sample from the Enshi prefecture in Hubei province. All lenses were graded and classified for opacities by slit lamp after papillary dilation, using the Lens Opacification Classification System II. The age-related cataract patients were 418 cases (33.28%). The prevalence of age-related cataract was 37.2% in women and 26.0% in men. The prevalence of nuclear cataract was 23.7%; cortical cataract was 22.4% and posterior subcapsule cataract was 5.2%. The prevalence of cataract of the 50–59 group was 13.41%; 60–69 group was 42.15%; 70 and over group was 61.9%. The prevalence of age-related cataract in high-selenium areas has not significantly increased; to some extent, the high selenium intake will not become a risk factor for the increase of cataract incidence.     

盐胁迫下不同树龄枸杞各器官的盐离子分布

以甘肃景泰沿黄灌区栽植的枸杞(Ly cium barbarum L.)为研究对象,研究了盐胁迫下不同树龄枸杞各器官的盐离子分配.结果表明,1~2年生枸杞各器官的K 分布为幼叶>成叶>嫩茎,而3~4年生枸杞为幼果>嫩茎>幼叶>成叶;1~4年生枸杞成叶的N a 含量分别为1.273%、1.414%、0.689%和0.497%;不同年龄枸杞成叶、幼叶、嫩茎和幼果的N a /K 平均值分别为2.425%、1.397%、0.489%和0.301%;随枸杞年龄的增加,根对土壤中N a 、K 的选择吸收比值(SA)逐渐增加.枸杞选择性吸收K 、拒N a 的特性和分配较多N a 、C l-给耐盐性较强的器官(成叶)是枸杞耐盐的重要原因.     

圆柏属常绿木本植物叶片抗氧化系统季节变化与抗冷冻的关系

测定分析圆柏属2种常绿木本植物叶抗氧化系统在冷冻适应过程中的季节变化。结果表明,叶能在组织结冰状况下生存与其具备完善的抗氧化保护系统有关,该系统能及时清除氧自由基、抑制膜脂过氧化、维持膜的完整性;冷适应期积累的活性氧可能诱导了抗氧化保护系统使叶片获得抗冻性。祁连圆柏的抗氧化系统比圆柏在抗冻性诱导中具有更广泛的适应策略。     

Influence of carrier proteins on the immunologic response to haptenic antitetrodotoxin vaccine

Tetrodotoxin (TTX) is a haptenic, highly toxic neurotoxin with no specific antidote available yet. Anti-TTX vaccine is being studied for antitoxin development. The effectiveness of the carrier protein in eliciting TTX-specific antibody response was comparatively studied. TTX was conjugated to Tachypleus tridentatus hemocyanin (TTH), Limulus polyphemus hemocyanin (LPH), tetanus toxoid (TT), diphtheria toxoid (DT), and bovine serum albumin (BSA) chemically to form artificial antigens TTH-TTX, LPH-TTX, TT-TTX, DT-TTX, and BSA-TTX, respectively, with which BALB/c mice were immunized, and the antibody response and antitoxic efficacy were detected. The serum anti-TTX antibody response and antitoxic efficacy varied markedly with adopted carrier protein. TTH-TTX elicited the best and BSA-TTX the worst TTX-specific antibody response. The proportion of the immunized mice surviving a 3x lethal dose (LD) dose of TTX challenge was 92%, 75%, 42%, 8%, and 0% for TTH-, TT-, LPH-, DT-, and BSA-TTX conjugates, respectively. The rank order of total efficacy of carrier protein for both anti-TTX antibody response and antitoxic effect was TTH > TT > LPH > DT > BSA. As a result of formaldehyde treatment in coupling of TTX carriers, the relative immunogenicity of TTX vs carrier, that is, the ratio of TTX- to carrier-specific antibody response, evidently varied with respective carrier adopted, in a rank order of TT > BSA > TTH > DT > LPH. The results suggest that the carrier protein used in haptenic TTX vaccine is greatly important in eliciting potent anti-TTX antibody, and both TTH and TT are the preferred carriers for development of excellent experimental TTX vaccine.     

Physiological Responses of Limonium aureum Seeds to Ultra-drying

The seeds of Limonium aureum （L.） Hill. were dried from 8.92% to 2.88% moisture content in a desiccating container with silica gel. After ultra-drying the seeds were accelerated aged （50 ℃, 1 month）, and some physiological indices, including the electrical conductivity, dehydrogenase activity, superoxide dismutase （SOD）, peroxidase （POD）, glutathione reductase （GR）, ascorbate peroxidase （APX）, catalase （CAT）, volatile aldehydes and malondialdehyde （MDA） were tested. The results indicated that dehydrogenase, POD, SOD, GR, APX and CAT activities of the ultra-dry seeds were higher than the control seeds, while volatile aldehydes and malondialdehyde were lower than the control group. The results suggest that ultra-drying is beneficial for maintaining the vigor of L. aureum seeds at a high level. Thus, L. aureum seeds could be stored under ultra-dry conditions.