Expression and characterization of Gag protein of HIV-1（CN） in Pichia pastoris

To express the core protein of HIV-1 of Chinese prevalent strain （HIV-1 （CN）） in Pichia pastoris, the fulllength gag gene was inserted into the secretory expression vector pHILS1. Linearized recombinant plasmid pHILGAG by Sail was electrotransformed into the yeast strain GS115, and the yeast transformants were identified by PCR. To induce the interest protein to be expressed, the PCR positive transformants were inoculated in the medium of BMGY and BMMY, mRNA of the strain was detected by RT-PCR, and the expressed protein was analyzed by SDS-PAGE, Western blotting and thin layer scanning. mRNA （1.3 kb） was amplified by RT-PCR. SDS-PAGE and Western blotting analysis showed that the molecular mass of the expressed protein was 55 kD, which was similar to the expected value, and the expressed protein could react with McAb to HIV-1 p24. Thin layer scanning analysis demonstrated that the whole amount of the expressed protein was approximately 13 % of the soluble protein in the supernatant. The recombinant yeast had good genetic stability. The optimal expression conditions of the engineering yeast were as follows： BMMY medium, 80-90% of dissolved oxygen, 1% methanol, and 3-day-cultivation course. Gag proteins were expressed under the optimal expression condition and purified via gel filtration chromatography. The purity of the interest protein was up to 85 %. After the purified proteins were inoculated into BALB/c mice, the anti-HIV-1 antibodies in the immunized mice could be detected by Western blotting.     

语言刺激的功能性MR成像在自闭症诊断中的潜在应用价值

目的对自闭症病人给予被动性语言刺激,评价采用功能性磁共振(MR)成像作为判断病人有无语言缺陷的客观指标的可行性。材料与方法本研究为前瞻性研究,研究方     

Changes in the configuration of peripheral blood erythrocytes in patients with pneumoconiotic tuberculosis and pulmonary tuberculosis during treatment

The impact of enterosorption on the morphology of peripheral red blood cells was examined in patients with coniotuberculosis and pulmonary tuberculosis by using scanning electron microscopy. Enterosorption was found to promote decreases in the proportion of abnormally changed cellular elements and increases in normal cells, which confirms the stimulating and antitoxic effects of this treatment.     

The influence of pretransplant lipoprotein abnormalities on the early results of renal transplantation

Abstract. Lipoprotein patterns were investigated before and after renal transplantation in a prospective study including 151 patients. Kidney graft losses during the first 6 months were associated with higher total cholesterol ( P = 0.03), LDL cholesterol ( P = 0.003) and LDL triglyceride levels ( P = 0.01) before transplantation. Patients with serum cholesterol ±6.9 mmol l^-1 before transplantation had more acute rejections (1.7 vs. 0.9), a worse graft function and more vascular intimal hyperplasia and glomerular mesangial changes in transplant biopsies at 6 months. Patients with serum creatinine levels exceeding 160 μmol l^-1 at 6 months had more severe lipid disorders already before transplantation.
Serum creatinine at 6 months was influenced by the number of acute rejection episodes ( P = 0.0001) and the age of the donor ( P = 0.009) while the number of acute rejections was found to be related to pretrans plant total cholesterol levels ( P = 0.0086) and the age of the recipient ( P = 0.025).
In conclusion, pretransplant lipoprotein disturbances have an impact on the early outcome of renal transplantation. Since there is a progresssion of hyperlipidaemia following transplantation, this may have an influence also on the cardiovascular morbidity and late graft dysfunction.     

酸中毒时心脏克隆钾离子通道Kv1．4的动力学特性

目的：探讨心脏克隆钾离子通道Kv1．4的C型失活(Kv1．4△N)在非洲蟾蜍卵母细胞上表达后的动力学特性以及酸中毒时的改变。方法：将Kv1．4△N cRNA(最大体积为50 n1)注入非洲爪蟾的卵母细胞内,于18℃孵育16h以上。电极采用两步法拉制,微电极由1．5 mm口径的电极拉制。电极内充3M KCl,电阻0．5～1．0MΩ。采用双微电极电压钳制法(two electrode voltage clamp,TEV)在室温下(20～24℃)记录电流。结果：与正常pH时相比,酸性环境下,Kv1．4△N的峰电流减小,在pH7．4时通道在去极化至-40mv激活,而pH 6．8时为-30mv激活；通道在pH 7．4时最大失活为0．384±0．072,而在pH6．8时为0．197±0．013；在pH6．8时通道复活减慢(P＜0．05)。结论：酸中毒导致通道电流减小,并且使通道失活加快和恢复减慢。     

Endoscopic ultrasonography guided biliary drainage: Summary of consortium meeting,May 7^th , 2011, Chicago

Endoscopic retrograde cholangiopancreatography (ERCP) has become the preferred procedure for biliary or pancreatic drainage in various pancreatico-biliary disorders. With a success rate of more than 90%, ERCP may not achieve biliary or pancreatic drainage in cases with altered anatomy or with tumors obstructing access to the duodenum. In the past those failures were typically managed exclusively by percutaneous approaches by interventional radiologists or surgical intervention. The morbidity associated was significant especially in those patients with advanced malignancy, seeking minimally invasive interventions and improved quality of life. With the advent of biliary drainage via endoscopic ultrasound (EUS) guidance, EUS guided biliary drainage has been used more frequently within the last decade in different countries. As with any novel advanced endoscopic procedure that encompasses various approaches, advanced endoscopists all over the world have innovated and adopted diverse EUS guided biliary and pancreatic drainage techniques. This diversity has resulted in variations and improvements in EUS Guided biliary and pancreatic drainage; and over the years has led to an extensive nomenclature. The diversity of techniques, nomenclature and recent progress in our intrumentation has led to a dedicated meeting on May 7 th , 2011 during Digestive Disease Week 2011. More than 40 advanced endoscopists from United States, Brazil, Mexico, Venezuela, Colombia, Italy, France, Austria, Germany, Spain, Japan, China, South Korea and India attended this pivotal meeting. The meeting covered improved EUS guided biliary access and drainage procedures, terminology, nomenclature, training and credentialing; as well as emerging devices for EUS guided biliary drainage. This paper summarizes the meeting’s agenda and the conclusions generated by the creation of this consortium group.     

Neurophysiological investigations of the features of the state and physiological activity of some structures of the striopallidum and thalamus in various forms of parkinsonism

The results of many years of investigations of the features of the functional state and physiological activity of some structures of the striopallidum (Cd, GP, Put) and thalamus (VP, VL, CM) as links in the brain systems of the organization of mental and motor activity in patients with various forms of parkinsonism are presented in generalized form. The possibilities for the use of various types of ultraslow physiological processes to study the brain mechanisms of the activational and emotional states, with more precise definition of the role of structures of the striopallidal complex in the formation of systems supporting these types of mental activity in parkinsonism, are reviewed. Laboratory of the Physiology of the States of the Brain and Organism, Institute of Human Brain, Russian Academy of Sciences, Saint Petersburg. Translated from Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 80, No. 1, pp. 78–87, January, 1994.     

Defect of epidermal 12(S)-hydroxyeicosatetraenoic acid receptors in psoriasis

12-hydroxyeicosatetraenoic acid (12-HETE) is assumed to play a central role in the pathophysiology of psoriasis. Since its effects in skin are mediated by specific high-affinity receptors, we studied the receptor characteristics in cultured epidermal cells from involved and apparently healthy skin of psoriasis patients by radioligand binding assay. Involved and uninvolved psoriatic epidermal cells showed a fourfold decrease in the number of 12-HETE binding sites as compared with normal healthy individuals and patients with atopic dermatitis, while receptor affinity remained unchanged. The decrease in receptor number was evident in psoriatic cells even in long-term culture and was not due to receptor down-regulation, defective response to interferon gamma or to protease degradation of receptor protein. The decrease in the number of 12-HETE receptors detectable even in clinically normal psoriatic skin functionally leads to diminished 12-HETE uptake and may thus represent a primary central molecular defect in the pathophysiology of the disease.