果胶酶亲和吸附剂的制备及其性能比较

以甜菜渣、果胶、海藻酸钠为原料 ,与表氯醇在碱性条件下交联 ,制备了果胶酶亲和吸附剂并应用这几种亲和吸附剂对草酸青霉果胶酶和黑曲霉果胶酶进行亲和层析实验。结果表明 :用小于 60目的甜菜渣粉末和海藻酸钠制备的果胶酶亲和吸附剂 (分别简写为SBP吸附剂和SA吸附剂 )具有较好的纯化果胶酶性能 ,其中SBP吸附剂性能稳定 ,而SA吸附剂膨胀系数较大 ,在流动相改变时引起流速不稳。     

A self-priming microfluidic diaphragm pump capable of recirculation fabricated by combining soft lithography and traditional machining

Fluid transport is crucial in the development of microanalytical devices. While there are many micropump designs available, most are incapable of sustaining recirculation of fluid at microL/min to mL/min levels. We have designed and fabricated a positive displacement micropump by combining soft lithography with traditional bulk machining. The micropump is actuated through pneumatic pressure. The pump is self-priming and is suitable for recirculating fluid through a microfluidic device containing mammalian cell culture. By custom designing the volume of the pumping chamber, tight control of the output flow rate can be obtained by changing the actuation frequency. It can also be fabricated easily on plastic substrates without access to expensive microfabrication equipment.     

奥氮平与碳酸锂分别联合丙戊酸钠治疗双相障碍躁狂发作 的临床疗效比较

目的:比较奥氮平与碳酸锂分别联合丙戊酸钠治疗双相障碍躁狂发作的临床疗效,探讨提高双相障碍躁狂发作临床疗效的药物治疗方案。方法:选择双相障碍躁狂发作患者90例,随机均分为A组与B组,A组给予奥氮平联合丙戊酸钠治疗,B组给予碳酸锂联合丙戊酸钠治疗,比较两组患者治疗第2周、第4周、第6周躁狂量表(BRMS)评分、副反应量表(TESS)评分和治疗第6周的临床疗效。结果:两组患者在上述方面比较,差异均具有统计学意义(P<0.05),A组临床疗效好于B组。结论:药物治疗双相障碍躁狂发作时,应选择奥氮平联合丙戊酸钠治疗方案,可提高临床疗效,减少用药后副反应。     

蚯蚓对土壤微生物残留物的影响研究评述与展望

蚯蚓如何影响土壤有机碳的固持是土壤生态学的关键科学问题之一。蚯蚓能同时促进土壤有机碳分解和稳定,这种两面作用带来的不确定性被研究者称为"蚯蚓困境"。研究证据和新兴的"土壤微生物碳泵"概念模型表明土壤微生物残留物是土壤有机质的主要贡献者。为系统了解蚯蚓对土壤微生物残留物的影响与可能的机制,研究分析和总结了已有的国内外蚯蚓与微生物残留物（氨基糖）的相关研究成果,表明：（1）过往的研究忽略了蚯蚓对微生物残留物的影响,导致这一方向的研究严重滞后;（2）蚯蚓对土壤微生物残留物影响的方向和大小仍有很大的不确定性,可供量化分析其驱动机制的研究还很缺乏。研究尝试将蚯蚓整合到"土壤微生物碳泵"概念框架中,分析蚯蚓影响土壤微生物残留物3个方面的可能机制,即：（1）改变土壤微生物量、群落结构,（2）改变微生物生理特性,（3）改变土壤团聚体结构等,影响土壤有机碳的积累。同时,本文提出了未来相关研究的6个重点方向,包括：（1）蚯蚓对微生物的选择性取食,（2）肠道介导的微生物"涨落"现象,（3）蚯蚓对矿质结合有机物的"破坏"与"重组",（4）蚯蚓引起的"激发"和"续埋"效应,（5）多生态型相互作用,（6）全球变化背景下的蚯蚓生态学等,以期为进一步揭示蚯蚓-微生物相互作用影响土壤有机碳累积与稳定性的机制提供参考。     

Subcellular distribution and immunocytochemical localization of Na,K-ATPase subunit isoforms in human skeletal muscle

Summary

The expression of Na,K-ATPase isoforms was investigated in human skeletal muscle membranes isolated by subcellular fractionation. The α1, α2, α3 and β1 subunits were detectable in membranes prepared from the human soleus muscle. The α1 subunit was largely detected in a fraction enriched with plasma membranes (PM), its abundance in an Intracellular membrane fraction (IM) accounted for only 4% of that in the PM fraction. No α1 subunits were detected in membranes of sarcoplasmic reticulum (SR) origin. The PM and IM fractions were enriched with α2 subunits which were less abundant in the SR-enriched fraction. The abundance of α2 molecules within the IM fraction was about 75% of that in the PM fraction when the total protein content for the two fractions was taken into account. Immuno-cytochemical studies confirmed the localization of the α1 subunit to the muscle cell surface. The α2 subunit was also found to be present in the cell surface but the observation that α2 immuno-fluorescence was diffusely dispersed throughout the muscle fibre indicated that it was also present intracellularly, consistent with its biochemical localization in the PM and IM membrane fractions. The α3 subunit was detected largely in the PM fraction but the lack of good antibodies to this isoform precluded an analysis of its immunocytochemical localization. The β1 subunit was enriched in the PM fraction but was also detected to a modest extent in the IM. A polyclonal anti-β2 antibody, which reacted positively with both human brain microsomes and rat skeletal muscle membranes, revealed that human skeletal muscles contained no immunoreactive β2 subunits. Our results indicate that the human soleus expresses the α1 and α2 (and possibly the α3) subunits of the Na,K-ATPase and that the activity of these isoforms must be supported by the β1 subunit in this muscle.     

Sodium Pentaborate Pentahydrate ameliorates lipid accumulation and pathological damage caused by high fat diet induced obesity in BALB/c mice

BackgroundObesity is one of the most popular topic in the field of research. In order to defeat this highly widespread disease, the mechanism of fat accumulation at the molecular level and its elimination are crucial. The use of boron has been showing promising results during the recent years.MethodsIn this study, anti-obesity potential of Sodium Pentaborate Pentahydrate (SPP) used as a dietary supplement on BALB/c mice fed with a high-fat diet was evaluated. Mice were divided into four groups with different diets, consisting of a normal diet, a high-fat diet (HFD) (containing 60 % fat), a HFD-supplemented with 0.5 mg/g body weight (BW) of SPP and a HFD-supplemented with 1.5 mg/g body weight (BW) of SPP. The animals were then observed for 10 weeks and physically monitored, and were sacrificed at the end of the experiment for physical and physicochemical evaluation.ResultsAccording to the physical parameters measured -body weight, food and water intake ratios-, the results indicate that SPP decreased weight gain in a dose dependent manner. Measurement of the hormone levels in the blood and fat accumulation in organs of mice also supported the anti-obesity effects of SPP. Expressions of adipogenesis related genes were also negatively regulated by SPP administration in white adipose tissue (WAT) tissue.ConclusionThese findings promise a treatment approach and drug development that can be used against obesity when SPP is used in the right doses. As a future aspect, clinical studies with SPP will reveal the effect of boron derivatives on obesity.     

Definition of Epitopes for Monoclonal Antibodies Developed Against Purified Sodium Channel Protein: Implications for Channel Structure

To test sodium channel structural models, we defined the epitopes for nineteen independently cloned monoclonal antibodies previously generated against purified, detergent-solubilized, adult rat skeletal muscle sodium channel protein using channel proteolysis, synthetic peptides, and fusion proteins. All identified epitopes were continuous and unique to the skeletal muscle subtype α-subunit. Of the nineteen independent clones, seventeen had epitopes located either in the origin of the amino-terminus or in the interdomain 2–3 region while only two antibodies had epitopes located in the mid-portion of the interdomain 1–2 region. No immunogenic regions were identified on the α-subunit's extracellular regions, interdomain 3–4 segment, or carboxyl-terminus or on channel β-subunits. While immune tolerance may explain the lack of immunogenicity of extracellular regions, the lack of immunogenicity of most of the channel's cytoplasmic mass may be due to segment inaccessibility from organization of these regions as globular domains, to insertion of parts of these regions into the membrane phase, or to interaction with other protein elements. The definition of monoclonal antibody epitopes allows us to reinterpret previously reported monoclonal antibody competition studies, providing independent support for our model of sodium channel cytoplasmic domain structure. In addition, these data suggest additional testable hypotheses concerning the interactions of the sodium channel amino- and carboxyl-termini with each other as well as with other protein elements. Received: 4 March 1998/Revised: 15 May 1998     

Structural and functional characterization of the human NBC3 sodium/bicarbonate co-transporter carboxyl-terminal cytoplasmic domain

The sodium bicarbonate co-transporter, NBC3, is expressed in a range of tissues including heart, skeletal muscle and kidney, where it modulates intracellular pH and bicarbonate levels. NBC3 has a three-domain structure: 67 kDa N-terminal cytoplasmic domain, 57 kDa membrane domain and an 11 kDa C-terminal cytoplasmic domain (NBC3Ct). The role of C-terminal domains as important regulatory regions is an emerging theme in bicarbonate transporter physiology. This study determined the functional role of human NBC3Ct and characterized its structure using biochemical techniques. The NBC3 C-terminal domain deletion mutant (NBC3ΔCt) had only 12±5% of wild-type transport activity. This low activity is attributable to low steady-state levels of NBC3ΔCt and almost complete retention inside the cell, as assessed by immunoblots and confocal microscopy, suggesting a role of NBC3Ct in cell surface processing. To characterize the structure of NBC3Ct, amino acids 1127–1214 of NBC3 were expressed as a GST fusion protein (GST.NBC3Ct). GST.NBC3Ct was cleaved with PreScission Protease? and native NBC3Ct could be purified to 94% homogeneity. Gel permeation chromatography and sedimentation velocity ultracentrifugation of NBC3Ct indicated a Stokes radius of 26 and 30 Å, respectively. Shape modelling revealed NBC3Ct as a prolate shape with long and short axes of 19 and 2 nm, respectively. The circular dichroism spectra of NBC3Ct did not change over the pH 6.2–7.8 range, which rules out a large change of secondary structure as a component of pH sensor function. Proteolysis with trypsin and chymotrypsin identified two proteolytically sensitive regions, R1129 and K1183-K1186, which could form protein interaction sites.     

Sodium chloride tolerance in strains ofEpidermophyton floccosum andEpidermophyton stockdaleae

The ability of strains of genusEpidermophyton to grow at different concentrations of NaCl (ranged from 0 to 10%) was studied. A reduction in the diameter of colonies was observed as the concentration of NaCl increased. A nearly complete absence of macroconidia were shown even at the lowest concentration assayed. The geophilic speciesE. stockdaleae tolerated higher concentrations of NaCl (> 7%) thanE. floccosum (< 3%).