Biotechnology is one of the important areas of life science, consequently the ethical problems were caused by its researches and applications. The article analyzed the ethical problems related to the development of biotechnology and put forwards some countermeasures in order to keep it developing constantly and steady.

OBJECTIVE:To investigate the role of clinical pharmacists in pediatric severe infection treatment,and to provide reference for rational drug use in clinic. METHODS:Based on related typical cases,the breakthrough points of clinical pharmacists providing pharmaceutical care in pediatric severe infection treatment plan were introduced. RESULTS:Clinical pharmacists formu-lated multi-drug resistance pathogenic antibacterial plan according to the results of pathogenic microbiological examination,use pharmacokinetics/pharmacodynamics,excluded interference to pediatric anti-infection treatment from ADR,and guideline and eco-nomical factors of patients. CONCLUSIONS:Clinical pharmacists adopt professional knowledge of pharmacy to provide pharmaceu-tical care in pediatric severe infection treatment and play professional assisted role so as to guarantee safe,effective and economical treatment.

Objective:Defensins from human neutrophils exhibit a broad antimicrobial activity.The transfection of the gene fused by human neutrophil peptide 1(HNP1) with the J chain into other cells may induce the in vitro expression of HNP1 to play antimicrobial roles.The authors analyzed the influencing factors on the expressed proteins of HNP1 and the J chain gene in COS-7 cells.Methods: Western blot was used to analyze the technical factors influencing the HNP1 protein expression.Results: Recombinant J-HNP1 was expressed in the cells successfully.Conclusion: The expression of J-HNP1 can be detected inside and outside the cells by Western blot.

Objective To investigate the effect of postoperative analgesia with oxycodone on T cell function after operative of cesarean section with chronic hepatitis B.Methods Sixty cesarean sec-tion women with chronic hepatitis B undergoing CS,aged 22-35,were randomly divided into two groups:oxycodone group (group O)and morphine group (group M).The changes of immune cells (Th1,Th2)and liver function were recorded after the analgesia (immediate,postoperative 24 h,48 h,72 h).The total number of pressing analgesia pump and the cumulative amount of PCA were re-corded.Results The Th1 of group O was higher than that of group M at 24 h,48 h after operation (P <0.05),while there was no significant difference of Th1 and Th2 in group M.The total patient-controlled pressing times and accumulated amount of PCA of group O were significantly lower than those in group M (P <0.05).There was no significant difference in the incidence of adverse reactions between the two groups.Conclusion Oxycodone can activate T cell function in postoperative analge-sia,while morphine causes the inhibition of Th1 cells.

BACKGROUND:Thoracolumbar fracture often accompanies with the injury of adjacent intervertebral disc. Traditional posterior short-segment fixation does not deal with the injured intervertebral disc, which may be the main reason for kyphosis in patients after surgery.
OBJECTIVE:To investigate the effect of injured intervertebral disc on kyphosis angle in patients with single vertebral thoracolumbar fracture after treated with posterior short-segment fixation alone.
METHODS:From January 2009 to June 2014, 40 cases of thoracolumbar fractures were treated in Jinan Central Hospital. They were folowed-up at preoperation, 2 and 12 months after operation and 6 months after internal fixation removal. Data were obtained from X-ray and MRI scanning. According to the preoperative MRI images, cases were assigned to observation group (17 cases) and control group (23 cases) according to injury and non-injury intervertebral disc. Data of vertebral wedge angle, sagittal plane kyphosis, proximal intervertebral disc angle, sagittal index and degeneration classification of proximal intervertebral disc angle from two different groups were analyzed at each folow-up time point (18-30 months, averagely 23.6 months).
RESULTS AND CONCLUSION:(1) Imaging parameters: sagittal plane kyphosis was significantly severer at 6 months than that at 2 months in both groups (P < 0.05). Sagittal plane kyphosis, proximal intervertebral disc angle and sagittal index were greater in the observation group than in the control group at 12 months after surgery and 6 months after fixator removal (P < 0.05). (2) Degeneration classification of proximal intervertebral disc angle: Pearce degeneration grade of proximal intervertebral disc was significantly higher in the observation group than in the control group at 2 months after surgery and 6 months after internal fixation removal (P < 0.05). (3) Results suggested that kyphosis may appear in the patients with thoracolumbar fracture after a posterior short-segment fixation alone, and the injured disc may lead to more severe kyphosis.

Objective To summarize our experience in diagnosis and treatment of splenic tumors.Methods A retrospective study of the clinicopathologic data of 92 cases of splenic tumor was performed.Results There were 47 benign and 45 malignant tumors.The preoperative confirmed diagnostic rate by B-US was 85.87%(79/92),by CT was 91.30%(84/92).Splenectomy was performed in 81 cases,tumor resection or partial splenectomy in 7 cases,needle aspiration of splenic cyst in 3 cases and biopsy in one case.The 1-,3-,5-,10-year survival rate of malignant tumor was 68.29%,31.70%,14.63%,0.24%,respectirely.Conclusions Imaging studies are the main diagnostic methods of splenic tumors.Splenectomy is the treatment of choice for primary splenic malignant tumor.Splenic benign tumor does not require any other therapy after operation.Splenic malignant tumor requires adjuvant treatment.

Objective To obtain the disulfide stabilized Fv fragment (dsFv) against N-terminal fragment of human lip polysaccharide binding protein (NH-LBP) and to identify its biological vitality. Methods The disulfide stabilized Fv fragment antibody (dsFv) was obtained after the inclusion bodies of dsFvVH and dsFvVL had been refolded and purified. Then the characteristics of dsFv were determined in vitro by ELISA and by detecting the secretion of tumor necrosis factor alpha (TNF-?) in rats. Results There was 2.1 mg protein of dsFv obtained. dsFv had good combination with NH-LBP and could restrain inflammatory reaction caused by lip polysaccharide (LPS) in vivo. Conclusion It is feasible to get dsFv against NH-LBP by respective expression of VL and VH. The partial inhibition of the biological function of LBP by dsFv is a new way to restrain the over-inflammatory reaction in vivo.

Objective To reconstruct the proteinic sequence of human cathelicidin LL-37 to increase the bactericidal activity of LL-37 and to express the reconstructed LL-37 (rLL-37) in bacterium. Methods The two dimensional structure, three dimensional structure and chemical characteristic of LL-37 were analyzed by Soft Ware Anthepro 5.0 and SWISS-MODEL. Without the three dimensional changes of LL-37, some negative amino acids of human cathelicidin LL-37 were replaced by positive amino acids and the positive charge of LL-37 was increased. According to the proteinic sequence changes of rLL-37, the DNA sequence of rLL-37 was reconstructed by Touch-Down PCR and recombined with vector pET-28a (+), thus rLL-37 was expressed in E.coli. BL21 (DE3) by the induction of IPTG and was purified by chromatography. Results Glu~ 16 , Asp~ 26 , Glu~ 36 of LL-37 were replaced by Gln~ 16 , Asn~ 26 , Gln~ 36 and the static charge of LL-37 was increased from +5.8 to +9.0 at pH 7.4. The DNA sequence of rLL-37 was reconstructed and inserted into vector pET-28a (+), the rLL-37 was expressed in E.coli. BL21 (DE3) and purified by strong cation exchange supports Macro-Prep High S successfully. The rLL-37 was proved by the means of inhibitory zone to be able to kill Gram-negative bacteria and Gram-positive bacteria. Conclusion It is feasible to reconstruct human cathelicidin LL-37 and express the protein in bacteria by fusion, which make it possible to produce more rLL-37 and study its biological function deeply.

Objective To obtain and identify monoclonal antibody against human lipopolysaccharide binding protein (LBP). Methods Three clones of antibody with better activity against human LBP were isolated after human antibody library screening by human LBP and 5 rounds of enrichment. After plasmid extracting, geneⅢ cutting, self-recircling and electric transforming, soluble Fab was expressed in E.coli. XL1-blue by the induction of IPTG and its characteristics were determined. Results The positive antibody was concentrated to 8.16?104 folds and the highest activity of the three clones was 106 . Conclusion Phage antibody against human LBP has been obtained successfully, which lays a solid foundation for researching its function.

Objective To employ 2 approaches to construct and express reconstructed LL-37 (rLL-37) in procaryotic system, and to explore a better preparation method. Methods The first method: the rLL-37 was inserted into vector pET-28a (+), then was induced to express in E.coli. BL21 (DE3) and purified by chromatography; the second method: the rare codons in the rLL-37 gene sequence were substituted by the preferred codons of procaryotic cell, and a fragment of carrier protein molecule (CPM) was added to the N termination of the objective sequence to construct expression plasmid pET-30a(+)-CPM-rLL-37, then the rLL-37 was expressed in E.coli. BL21 Star(DE3) and purified by chromatography. The productive rates of the 2 methods were compared and the antimicrobial effects of obtained rLL-37 was studied. Results The first method: the DNA sequence of rLL-37 was obtained successively by Touch-Down PCR. The expression plasmid pET-30a(+)-CPM-rLL-37 was expressed with fusion protein in E.coli BL21 (DE3). The expression rate accounted for 20% of total bacterio-protein, then the expressed product was purified by using high positive ion exchange column Macro-Prep High S; The second method: a fragment of carrier protein molecule was designed that contained 28 amino-acid residue and its pHi was 2.7, net charge was-6.0 at pH 7.4. After the expression plasmid pET-30a(+)-CPM-rLL-37 was constructed successively, it was expressed in E.coli BL21 Star (DE3). The expressed fusion protein accounted for 35% of total bacterio-protein, then the expressed product was purified by using affinity binding chromatography with TALON resins successfully. The obtained 2 kinds of rLL-37 were able to kill both Gram-negative and-positive bacteria by the means of inhibitory zone. Conclusion It’s feasible to prepare efficiently rLL-37 in procaryotic system, which founds the basis for the further research on bactericidal activity of rLL-37.