壳聚糖酶的克隆表达与壳寡糖的制备分析

目的:克隆壳聚糖酶基因于大肠杆菌中实现高表达,制备壳寡糖。方法:以枯草芽孢杆菌总DNA为模板扩增壳聚糖酶基因(CSN),克隆至载体pET23a(+)上,转化菌株BL21(DE3)。重组子经0.5 mmol/L IPTG诱导后,SDS-PAGE和质谱检测与鉴定重组酶。酶纯化后水解壳聚糖,薄层色谱分析其水解产物。结果:质谱证明壳聚糖酶(31.5kDa)成功表达,表达量占菌体总蛋白的45%左右。纯化后重组酶浓度为900 mg/L,纯度95%、回收率85%,酶活力为10 000 U/mg。壳聚糖降解产物为壳二糖至壳四糖。结论:原核表达载体pET23a(+)-CSN构建正确,壳聚糖酶表达量与活性高,适用于水解壳聚糖制备壳寡糖。     

Obstetrics and gynecology: AIDS and pregnancy

The Scientific Board of the California Medical Association presents the following inventory of items of progress in obstetrics and gynecology. Each item, in the judgment of a panel of knowledgeable physicians, has recently become reasonably firmly established, both as to scientific fact and important clinical significance. The items are presented in simple epitome and an authoritative reference, both to the item itself and to the subject as a whole, is generally given for those who may be unfamiliar with a particular item. The purpose is to assist busy practitioners, students, research workers, or scholars to stay abreast of these items of progress in obstetrics and gynecology that have recently achieved a substantial degree of authoritative acceptance, whether in their own field of special interest or another.The items of progress listed below were selected by the Advisory Panel to the Section on Obstetrics and Gynecology of the California Medical Association and the summaries were prepared under its direction.     

Pleasure and disgust: memories and associations of pleasant and unpleasant odours in Germany and Japan

The question of whether odour experiences have a function inthe cultures of the technically industrialized world was investigatedby interviews with German (166) and Japanese (88) subjects.They were asked to name pleasant and unpleasant odours frommemory and to give associations regarding them. The participantsenumerated 2040 odours and 3520 associations. Although Japanesesubjects in general gave less information than the Germans,the results for both cultures were very similar in quality aswell as quantity. The odour memories included the whole physicaland social environment and the associations showed the far-reachingeffect of odour experiences. Both pleasant and unpleasant odourswere remembered to an equal extent. Most of the odours recalledwere assessed similarly by the subjects, as either pleasantor unpleasant. Obviously, individual likings or aversions aredominated by underlying evaluations common to all subjects ofboth cultures. Uniformly judged odours were, for example, odoursof plants (pleasant) and odours of rotten and decomposed things(unpleasant). It is discussed whether this finding reflectspreprogrammed survival strategies. The few cultural differenceswhich were found seemed to reflect different life habits, normsand values in the two countries.     

人血管能抑素基因的克隆、表达及其表达产物的纯化和生物活性测定

以人胎盘脐带组织为材料,提取组织总RNA,用netRTPCR方法合成人血管能抑素cDNA基因,将该cDNA克隆进pSP72载体获得重组质粒pSP72C, DNA序列分析结果与预期序列一致。用BamHⅠ和NdeⅠ双酶切,切下pSP72C上的血管能抑素cDNA,插入pET3c载体的相应位点获得重组表达质粒pETC, 转化E. coli BL21(DE3), SDSPAGE分析显示：在IPTG诱导下,血管能抑素基因获得了高效表达,表达量约占菌体总蛋白的 27.9 %,主要以包涵体形式存在。包涵体经过洗涤、裂解、蛋白复性以及Sephadex G75凝胶过滤层析等步骤后,获得了纯度达91.4 %的人血管能抑素。CAM实验证明10 μg纯化蛋白就能显著抑制鸡胚新生血管生成。     

Astrocytes and Synaptosomes Transport and Metabolize Lactate and Acetate Differently

Astrocytes transport the monocarboxylate acetate, but synaptosomes do not. The reason for this is unknown, because both preparations express monocarboxylate transporters (MCT). The transport and metabolism of lactate, another monocarboxylate, was examined in these two preparations, and the results were compared to those for acetate. Lactate transport is more rapid in astrocytes than in synaptosomes, but of lower affinity (Kms of 17 and 4 mM, respectively). Lactate (0.2 mM) is metabolized to CO2 more rapidly in synaptosomes than in astrocytes (rates of 0.37 and 0.07 nmol x mg protein(-1) x min(-1), respectively). The reason for this is unclear, but cellular differences in lactate dehydrogenase isotype expression may be involved. Acetate is metabolized to CO2 more rapidly in astrocytes than in synaptosomes (rates of 0.43 and 0.02 nmol x mg protein(-1) x min(-1), respectively). This is likely due to cellular differences in the expression of monocarboxylate transporter subtypes.     

Knowledge and opinions about tissue donation and transplantation among Polish students and physicians

The idea of transplantation seems to be commonly identified by lay and professional people only with transplantation of vascularized organs like kidney or heart. The question arises whether there is any awareness of tissue transplantation among the public. A survey was therefore undertaken to assess awareness of and approaches to tissue donation and transplantation among selected social groups. A questionnaire on donation and transplantation issues was administered to respondents from the following groups: secondary school students, non-medical university students, medical university students, physicians. On the whole, 441 non-randomly sampled respondents were surveyed. The awareness of tissue transplantation is narrower than the awareness of organ transplantation. The support for tissue transplantation is weaker than for organ transplantation. This study shows that there is an acute need for education in the legal aspects of transplantation and that ways of motivating healthcare professionals to promote transplantation should be developed.     

Liquid chromatographic assay of phenothiazine, thioxanthene and butyrophenone neuroleptics and antihistamines in blood and plasma with conventional and radial compression columns and UV and electrochemical detection

An assay strategy for determining a wide range of phenothiazine, thioxanthene and butyrophenone neuroleptics and antihistamines both alone and in combination in blood and plasma is described. The general method employs liquid chromatography with both conventional and radial compression nitrile bonded columns. Detection is by ultraviolet absorption spectrophotometry or by amperometry depending on the concentrations to be measured. Ultraviolet absorption is suitable down to 10 ng/ml. Below this level amperometry is preferable. The various compounds are used as internal standards for each other. The lower limit of detection is approximately 0.1 ng ml⁻¹ with a 10-ml sample. The within-run coefficient of variation is a maximum of 7.3%.     

Protein synthesis by isolated etioplasts and chloroplasts from pea and wheat and the effects of chloramphenicol and cycloheximide

Etioplasts capable of incorporating ¹⁴C-leucine into protein have been isolated from dark-grown pea and wheat plants. The requirements for leucine incorporation for etioplasts were similar to those for chloroplasts. An ATP-generating system, Mg²⁺, and GTP were required. The amino-acid-incorporation activity of etioplasts from wheat was comparable to that of chloroplasts on an RNA basis, whereas the activity of pea etioplasts was about 50% of the activity of pea chloroplasts. The incorporation of leucine into protein by etioplasts and chloroplasts from pea and wheat was inhibited by chloramphenicol, and to a slight extent by cycloheximide.     

Electronic and spatial structure of dimethyl orthophosphate and orthophosphate

The applicability of different variants of the semiempirical CNDO method to calculate the electronic structure of different conformations of all possible ionic forms of dimethyl orthophosphate and orthophosphate is considered. The CNDO/BW (Boyd-Whitehead parameterization) approximation with selected parameters for the P-O bond is shown to provide the best qualitative and sometimes quantitative agreement with ab initio methods. The dependences of energy and P-O bond strengths in P-O-CH₃ (P-O-H) groups on torsion angles at P-O bonds are obtained in this approximation for dimethyl orthophosphate and orthophosphate. The rotation of these groups is found to stremgthen one P-O bond stronger and labilize another. The energy minima of dimethyl orthophosphate and orthophosphate anions are shown to correspond to conformations where the strengths of the studied P-O bonds are almost the same, i.e., none of the bonds is weakened to a minimum. Protonation of these compounds increases the strength of P-O bonds and decreases the dependence of bond strength on torsion angles. The di-and trianions of dimethyl orthophosphate and orthophosphate are also studied. The growth of negative charge is shown to progressively weaken the P-O bond. The dependence of bond strength on torsion angle for the dianion is less pronounced than that for the monoanion. Calculation results are compared with experimental data known from literature. The significance of the data obtained for revealing essential features of the enzyme cleaving and forming the P-O bond is discussed.     

Cinnamoyl- and hydroxycinnamoyl amides of glaucine and their antioxidative and antiviral activities

The aporphine alkaloid glaucine has been converted into 3-aminomethylglaucine and its free amino group has been linked to cinnamic, ferulic, sinapic, o-, and p-coumaric acids. The antioxidative potential of the synthesized amides was studied against DPPH(*) test. All of the tested compounds demonstrated higher radical scavenging activity than glaucine and 3-aminomethylglaucine, and lower antioxidative effect than the free hydroxycinnamic acids. The newly synthesized compounds were tested in vitro for antiviral activity against viruses belonging to different taxonomic groups.     

Ligand binding and stabilization of malate- and lactate dehydrogenase

Binding of coenzymes, coenzyme fragments and phenolate ligands to malate- and lactate dehydrogenase was studied. From linear competition in titration experiments, the coenzyme binding site was concluded to bind all the ligands employed. The analogy between the phenolate ligands and tetraiodofluorescein which is known to bind at the adenosine binding site suggests binding of phenolates at this site. Coenzymes and coenzyme fragments retard the irreversible thermal inactivation of the enzymes. The retardation effect decreases in the order NADH greater than NAD greater than ADPR greater than or equal to AMP for both enzymes. Phenolate ligands binding to the adenosine pocket do not stabilize the enzymes. The stabilization is concluded to originate from the interaction of coenzyme phosphate and nicotinamide with the enzymes. The interactions with the adenosine moiety and with the second ribose seem to be ineffective in retardation of thermal denaturation.     

Synthesis and characterization of random and block copolypeptides derived from gamma-methylglutamate and leucine N-carboxyanhydrides

The synthesis of random and block copolypolyeptides derived from gamma-methylglutamate and leucine N-carboxyanhydrides using Al-Schiff's base complexes and allylamine as initiators is here reported. The copolymer structures were confirmed by (1)H and (13)C NMR. The calculation of the statistical average block lengths reveals the presence of longer methylglutamate units in the copolymer. The determination of the reactivity ratios indicated a slightly higher reactivity of gamma-methylglutamateNCA as compared to leucineNCA. Block copolypeptides containing glutamate and leucine units were obtained by sequential polymerization of the two NCAs using Al-Schiff's base complexes or allylamine in dioxane as solvent. Based on (13)C NMR spectra of copolymers exhibiting two signals corresponding to peptide linkages, we confirmed the block structure and concluded that the copolymerization proceeds by attack of an amino group present on a glutamate chain end onto a LeuNCA. The copolymerization with allylamine was also shown, from calculation of the average block lengths of sequences, to exhibit living behavior. Viscometry analysis further showed that molar masses of the copolypeptides obtained with Al-Schiff's base were quite close to those derived from allylamine, supporting the proposed mechanism of copolymerization.     

Conjectures and reveries

The origin of photosynthesis is speculated to have involved carbon dioxide and self-replicating iron-rich clays. The later evolution of photosynthesis is considered to have undergone four distinct phases: (1) The photoreduction of carbon dioxide by ferrous ion to form oxalate and formate. (2) The entry of sulfur into the evolving clay system which led to the formation of acetyl thioesters. The polymerization of the acetyl thioeters led to the formation of quinones. The formation of Fe₂S₂ and Fe₄S₄ cores appeared in this phase. (3) The ability to fix nitrogen characterized the third phase. This led to the formation of pyrrole, flavin, nicotinamide, phycobilins, porphyrins and chlorophyll. (4) Finally, phosphate entered the evolving system.The chromophores evolved from ferrous ion through the quinones, carotenoids, phycobilins to chlorophyll. This evolution of chromophores implies that photosynthesis began in the UV and evolved through the blue, yellow, orange into the red. The electron transport chain evolved from ferrous ion through the Fe₂S₂ and Fe₄S₄ cores to the hemes.     

Metabolism and action of insulin and glucagon in goat during lactating and dry period

The metabolism and action of insulin and glucagon were investigated in goats during mid lactating (50 days postpartum) and during the dry period. The animals were fed hay and concentrate during lactation (1:1) and only hay during dry period. Pulse doses of unlabelled insulin and glucagon were injected intravenously. The disappearance of insulin from the circulation was faster during lactation than during dry period; the metabolic clearance rate of insulin was significantly increased during lactation. In contrast, the kinetic parameters of glucagon disappearance were very similar during the two periods. Basal plasma hormones (i.e. before hormone injection) were higher during lactation than during dry period; the molar ratio insulin:glucagon was left unchanged. The increase in plasma insulin following glucagon-stimulated hyperglycaemia was similar during the two periods. The ability of insulin to elicit a decrease in blood glucose was markedly impaired during lactation when compared to dry period. In contrast the ability of glucagon to increase blood glucose was slightly improved during lactation. Those endocrine changes could be related to the effect of both lactation and diet.     

Structure and interactions of desmosomal and other cadherins.

The cadherin superfamily of cell-cell adhesion molecules is now known to include proteins of the desmosome as well as of the adherens type of junction. The desmosomal cadherins consist of two families of proteins, the desmocollins and the desmogleins, both of which are represented by different isoforms which are differentially expressed in epidermis. The desmocollins are quite similar to the classic cadherins in overall structure, but with alternatively spliced variants; the desmogleins have extra cytoplasmic sequences added onto the basic cadherin structure. The cytoplasmic domains are specialized for binding to 'mediator' proteins, such as plakoglobin, which interconnect to the intermediate filament system rather than the actin filaments as do the classic cadherins.     

Physical Anthropology and the dental and medical specialties

The rise of two sub-specialties in Physical Anthropology traces back to the Anatomy Departments of Schools of Medicine in Germany and France during the nineteenth century. The study of human diversity in bones and bodies was largely by medically-trained anatomists. There developed Medical Anthropology and Dental Anthropology, employing osteometry and craniometry on the skeleton, somatometry and cephalometry on the living body. As a result cross-sectional studies gave way to longitudinal studies and X-ray techniques were added to purely mensurational procedures. In Medical Anthropology the specialties most directly concerned are pediatrics, plastic surgery, endocrinology, and orthopaedics. In Dental Anthropology the specialties most directly concerned are pedodontics, orthodontics, oral surgery, and prosthodontics. The contributions of Physical Anthropology to each is discussed.     

蚊虫叮咬止痒消炎剂的研制及应用

目的研制蚊虫叮咬止痒消炎剂,观察应用该蚊虫叮咬止痒消炎剂对吉林洮南地区背点伊蚊和刺扰伊蚊叮咬后的治愈效果。方法药品制备采用渗漉法和超声波提取法,质量控制依据中华人民共和国国家标准《一次性使用卫生用品卫生标准》GB 15979-2002,效果观察采用局部外用涂抹法。结果该止痒消炎剂12、24和48 h平均痊愈率分别为24.30%、55.14%和17.76%,总有效率为97.20%。结论该蚊虫叮咬止痒消炎剂符合中华人民共和国国家标准《一次性使用卫生用品卫生标准》GB 15979-2002的规定,对吉林洮南地区优势蚊种叮咬后具有明显的治愈效果,适用于部队野外执行任务携带。