Objective: To clone and express Treponema pallidum (TP) specific antigens P15 and P47 and use them in clinical examination. Methods: P15 and p47 genes synthesized were inserted into a GST fusion expression vector. The recombinant antigens were purified by affinity chromatography and then coated on microplates to establish an ELISA kit. Using this kit, national TP standards, sera of normal persons and patients with TP and other diseases were tested. Results: The synthesized p15 and p47 genes sequenced were identical with those of GenBank. National TP standards were tested by ELISA kit which coated with these recombinant antigens, and the results accorded with the standards. High sensitivity and specificity was showed when 2 recombinant antigens were used in ELISA to detect the sera of normal persons and patients with TP and other diseases. Conclusion: The recombinant TP specific antigen P15 and P47 are suitable for establishment of ELISA kit in clinical examination.

Medical education of PhD candidates is of great importance to medical personnel training in our country. With the development of society, the progress of science and technology and renewal of knowledge, cultivation of innovative talents and improvement of graduate education quality is priority nowadays. This paper reflects on the progress we’ve made on medical education of PhD candidates. Though efforts have been made to improve the innovative ability of talents in our country gradually, we still need to be more innovative, more consistent and more scientific in the future. According to the state Degree Requirements and our goal of talent training, we should combine courses with scientific research systematically, pay more attention to scientific methodology and academic training, finally form a better training mode of PhD candidates majoring in medical science.

Human kallikrein-1 (hK1) gene was cloned from kidney tissues cDNA, it was inserted into the plasmid pPICZalphaA, then the yeast expression vector pPICZalpha-hK1 was constructed. After transformed into Pichia pastoris host X33, high-level expression transformants were screened by escalating the concentration of Zeocin (from 500 to 700 microg/mL) of YPD plate and medium. When temperature was 30 degrees C, pH 6.0 with induction duration of 64 hours in the 30 L fermenter, the highest yield can reach about 6500 u/L (1.25 g/L). The variation of glycosylation resulted in two kinds of molecules, i.e. rhK1-H with a heavy molecular weight and rhK1-L with a light one. rhK1 was purified from the supernatant through Phenyl hydrophobic interaction, Cu(2+)-charged Chelating and Anion-exchange chromatography. 0.28 g rhK1-H and 0.62 g rhK1-L can be purified from one liter supernatant. The yield recovery was 72% with a purity of > 96%. So far our yield of rhK1 is superior than known recombinant expression method reported by other researchers.
Amino Acid Sequence ; Base Sequence ; Chromatography, Ion Exchange ; methods ; Genetic Vectors ; genetics ; Humans ; Kidney ; metabolism ; Molecular Sequence Data ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Tissue Kallikreins ; biosynthesis ; genetics

Objective To evaluate the efficacy of ultrasound-guided great auricular nerve (GAN)block combined with general anesthesia for middle ear microsurgery in adult patients.Methods Sixty-four adult patients of both sexes,aged 18-60 yr,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,scheduled for elective middle ear microsurgery,were assigned into 2 groups (n =32 each) using a random number tabte:GAN block combined with general anesthesia group (group GANB+G) and general anesthesia group (group G).Patients received GAN block with 0.25％ ropivacaine 2 ml using ultrasoundguided in-plane technique before anesthesia induction in group GANB+G.Anesthesia induction and maintenance protocols (IV infusion of propofol and target-controlled infusion of remifentanil) were similar in two groups.Analgesia was performed with sufentanil,flurbiprofen axetil or parecoxib sodium after operation to maintain visual analog scale score ＜4 within 48 h after operation.The infusion duration and consumption of propofol and remifentanil,requirement for postoperative analgesia and occurrence of postoperative nausea and vomiting were recorded.Results Compared with group G,the consumption of remifentanil was significantly reduced,and the requirement for postoperative analgesia and incidence of postoperative nausea and vomiting were decreased in group GANB + G (P ＜ 0.05 or 0.01).Conclusion Ultrasound-guided GAN block combined with general anesthesia provides better analgesic efficacy in the the perioperative period in adult patients undergoing middle ear microsurgery.