Increment of hFIX expression with endogenous intron 1 in Vitro

This paper probes into the feasibility of increasing expression level of hFIX gene with endogenous intron 1 sequence.hFIX minigene was obtained with middle sequence truncated intron 1 inserted into the relative site of hFIX cDNA,and plasmid vector pKG5i‘IX,retroviral vector G1NaCi‘IX were constructed.These vectors were transduced into target cells of PA317,C2C12,primary rabbit skin fibroblasts (RSF) and primary human skin fibroblasts (HSF).The expression level of mixed colonies are PA317/pKG5i‘IX,151 ng/10^6 cells/24h;PA317/G1NaCi‘IX,308 ng/10^6 cells/24 h;C2C12/G1NaCi‘IX,186 ng/10^6 cells/24 h;RSF/G1NaCi‘IX,1929 ng/10^6 cells/24 h;HSF/G1NaCi‘IX,1646 ng/10^6 cells/24 h.These results indicated that hFIX minigene with intron l is able to increase the expression level to about 3 times of that of hFIX cDNA.Meanwhile,in order to study the application of hFIX minigene in the retroviral-mediated gene transfer system and refrain from intron splicing during viral production,a retroviral vector G1NaCi‘IXR with reversely inserted hFIX minigene expression cassette was constructed.The expression level of reverse constructor in PA317 cells was 390 ng/10^6 cells/24 h with 79% of bioactivity.PCR detection of HT/G1NaCi‘IXR cells infected with PA317/G1NaCi‘IXR supernatant confirmed the existence of intron 1 sequence.These results suggested that expression vector with forward-inserted intronl-carrying hFIX expression cassette can be used in directed gene transfer,but when using the retroviral-mediated gene transfer system,reversely-inserted intronl-carrying hFIX expression cassette should be considered.     

Ibuprofen Blunts Ventilatory Acclimatization to Sustained Hypoxia in Humans

Ventilatory acclimatization to hypoxia is a time-dependent increase in ventilation and the hypoxic ventilatory response (HVR) that involves neural plasticity in both carotid body chemoreceptors and brainstem respiratory centers. The mechanisms of such plasticity are not completely understood but recent animal studies show it can be blocked by administering ibuprofen, a nonsteroidal anti-inflammatory drug, during chronic hypoxia. We tested the hypothesis that ibuprofen would also block the increase in HVR with chronic hypoxia in humans in 15 healthy men and women using a double-blind, placebo controlled, cross-over trial. The isocapnic HVR was measured with standard methods in subjects treated with ibuprofen (400mg every 8 hrs) or placebo for 48 hours at sea level and 48 hours at high altitude (3,800 m). Subjects returned to sea level for at least 30 days prior to repeating the protocol with the opposite treatment. Ibuprofen significantly decreased the HVR after acclimatization to high altitude compared to placebo but it did not affect ventilation or arterial O₂ saturation breathing ambient air at high altitude. Hence, compensatory responses prevent hypoventilation with decreased isocapnic ventilatory O₂-sensitivity from ibuprofen at this altitude. The effect of ibuprofen to decrease the HVR in humans provides the first experimental evidence that a signaling mechanism described for ventilatory acclimatization to hypoxia in animal models also occurs in people. This establishes a foundation for the future experiments to test the potential role of different mechanisms for neural plasticity and ventilatory acclimatization in humans with chronic hypoxemia from lung disease.     

康氏木霉纤维素酶系中C_x酶多分子型的分离纯化及某些性质

从康氐木霉(Trichoderma k(?)ningii)白色变异株As 3.4001的粗酶制剂中,获得了纤维素酶系中的一组C_x酶(C_(x1) C_(x2) C_(x3) C_(x4))。分离步骤包括Sephadex G-75凝胶过滤,DEAESephadex A-50离子交换层析,ConA-Sepharose亲合层析,SE-Sephadex C-50离子交换层析及聚丙烯酰胺凝胶电泳。C_(x1)与C_(x2)的分子量不同而所带电荷相同,它们的分子量各自为44,500和34,000。C_(x2)—C_(x4)的分子量相同而所带电荷不同。纯化的C_(x1)—C_(x4)经聚丙烯酰胺凝胶电泳鉴定为单带。比较它们对羧甲基纤维素钠(CMC-Na)的糖化力及液化力表明在作用方式的随机性上C_(x2)>C_(x3)>C_(x1)>C_(x4)。     

Excitatory effect of tuftsin tetrapeptide on the activity of white rats

Studies on male white rats have shown that tuftsin (trelys-pro-arg) enhances the locomotion in animals as disclosed by a series of the behavioral tests. The effect is dose-dependent: a dose of 50 microgram/kg did not change any of the test parameters, while that of 150 microgram/kg induced a short-term elevation of locomotion measured with an "Animeks". Administration of tuftsin in a dose of 300 microgram/kg led to the enhancement of the animals' locomotion as measured with the "Animeks", to the increased "open field" running time and to the reduced latent period of the reaction during the training in a T-shaped maze. Also, this dose of the peptide relaxed the reactions associated with fear. It is assumed that the effects observed are consequent on the stimulant action of tuftsin on the body of white rats.     

Use of PVA-cryogel entrappedCitrobacter intermedius cells for continuous production of 3-fluoro-L-tyrosine

Summary The cells ofCitrobacter intermedius (containing L-tyrosine phenol lyase which catalyses the synthesis of 3-fluoro-L-tyrosine fromo-fluorophenol, pyruvate and ammonia) were entrapped into 10% poly(vinyl alcohol) cryogel beads. Cryogenic immobilization resulted in a moderate increase of the synthetic activity of the cells. During continuous operation for one month in a packed bed column reactor the productivity of the immobilized biocatalyst decreased by 30%.     

Functional-morphologic characteristics of the endocrine system of the progeny of adrenalectomized rats

The authors observed signs of premature maturation and activation of the hypophysio-adrenocortical system at the early postnatal period (the 70--90th hour) in the progeny of rats adrenalectomized 1 to 3 and 7 to 9 days before labour. A more pronounced in comparison with control (progeny of the sham-adrenalcetomized and intact rats) response to stress pointed to the early maturation of the suprahypophysial control mechanisms as well. Signs of functional exhaustion in the hypophysio-adrenocortical system and also in the thyroid gland were revealed in mature (6--10-month-old) rats born of adrenalectomized rats.     

Modification of Bielschowsky's method for detecting senile plaques and intracellular neurofibrils

The distinction of the modification of Bielschowsky's method lies in additional colouring of silver-golden sections with strong solutions of stable cresyl-violet and acetic acid, as well as in recommendations to use precise-profile cortical sections. The given modification helps to obtain preparations with definite orientation and guaranteed identification of different nerve tissue elements. The preparations obtained can be employed for quantitative analysis and for a detailed study of thin specific structures and cytologic properties of pathologically changed nerve and glial cells.     

马铃薯糖转运蛋白基因的克隆及表达分析

植物SWEET基因家族是一类糖转运蛋白,在植物的生理活动和生长发育过程中发挥着重要功能。为了解马铃薯SWEET基因的相关信息,探究其在马铃薯不同组织以及在生物胁迫与非生物胁迫下的表达特性。该研究采用同源克隆技术从马铃薯‘青薯9号’中克隆了StSWEET5基因(GenBank登录号为MN295671),其CDS序列长度为717 bp,编码238个氨基酸。系统进化树分析结果表明,StSWEET5与番茄的氨基酸序列相似性最高(97.06%)。qRT-PCR分析表明:StSWEET5基因在马铃薯各组织(根、茎、叶、花、块茎、匍匐茎)中均有表达,且在花中的表达显著高于其他组织;糖胁迫下,StSWEET5基因在根、茎、叶中均有表达,尤其在根中的表达差异最为显著(P0.05)。在晚疫病菌(Phytophthora infestans)诱导后36 h时,表达量达到最高,随后急剧下调。推测StSWEET5基因参与了马铃薯糖胁迫以及响应了晚疫病诱导的过程。     

Evaluation of the prevalence and economic burden of adverse drug reactions presenting to the medical emergency department of a tertiary referral centre: a prospective study

Background  

Adverse drug reactions (ADRs) are now recognized as an important cause of hospital admissions, with a proportion ranging from 0.9–7.9%. They also constitute a significant economic burden. We thus aimed at determining the prevalence and the economic burden of ADRs presenting to Medical Emergency Department (ED) of a tertiary referral center in India     

疣柄牛肝菌属一中国新记录种

报道了采自内蒙古呼伦贝尔市的中国疣柄牛肝菌属1个新记录种,即假褐疣柄牛肝菌Leccinum pseudoscabrum(Kallenb.)utara.。主要特征为菌肉伤后变红到紫褐色,再到黑褐色,基部菌丝体手触后呈紫褐色;具泡状菌丝,呈栅栏状。比较和讨论了该种与相近种在伤后不同的变色反应及菌丝形态。研究标本存放于吉林农业大学菌物标本馆(HMJAU)。