The major challenge in the development of recombinant biologics lies in generating and isolating rare high-producing stable single clone in a short period of time. The selection marker is an essential component of the plasmid vector, it plays an important part in the generation and screening of producing cell lines. Engineering the selection marker to enhance the stringency of selection for high producing cells is one of the most effective approaches to improve the cell line development process. Here, using Chinese hamaster overy (CHO) cells as an example, we introduce the application of selection marker for generation of recombinant biologics producing mammalian cell lines, methods of engineering the selection markers to enhance the selection stringency, and propose considerations on cell substrate stability and selection marker safety, in order to provide references for high-efficiency development of recombinant biologics.

To explore the feasibility of stable expression of Hantavirus H8205 strain G1 segment and human IL-2 fusion gene in Vero cells, and to examine the immune protection effects on mice vaccinated with this recombinant eukaryotic expression vector containing Hantavirus G1 gene and IL-2 gene. With the help of lipofectamine, the Vero cells were transfected with pcDNA3.1/HisB-IL-2-G1 and the positive cells were selected by G418. IFAT and SDS-PAGE electrophoresis were used to determine the stable transfection and expression of recombinant protein. Each mouse was inoculated with plasmids intramuscularly (i.m.) three times, 2 boosts were given at 2-week intervals, serum anti-hantavirus antibodies were detected by ELISA and neutralizing antibodies (NAb) were detected by Plaque Reduction Neutralization Test. The fusion protein expressed in Vero cells was 78 kD, corresponding to the estimated molecular size. The neutralizing antibody titers of mice with pcDNA3.1/HisB-IL-2-G1 were 1:20-1:80. IL-2/G1 fusion gene could be transferred in Vero cells and stably express the fusion protein. Specific humeral immune responses in mice can be induced with the recombinant eukaryotic expression vector containing the fusion gene, which lays the foundation for further development of therapeutic HTNV vaccine.

Objective To evaluate the effectiveness of discharge planning and home care for patients with peritoneal dialysis (PD).Methods A total of 60 PD patients were randomly divided into the intervention group and control group,30 cases in each group.Discharge planning and home care were carried out in the intervention group, while routine nursing and telephone follow -up service after discharge were implemented in the control group.The patientsˊscores of Zung Self -Rating Anxiety Scale(SAS),Self -Rating Depression Scale(SDS),Self management ability of patients undergoing peritoneal dialysis,complication rate and readmission rate were compared between the two groups at the next day admission,discharge,lth month and 3th after discharge.Results The scores of SAS and SDS in the intervention group were significantly lower than that in the control group at 3th month after discharge,the difference was statistically significant(tSDS =5.263,tSAS =3.812,P <0.05).The Self management ability of patients undergoing peritoneal dialysis was significantly higher than that of the control group at lth and 3th month after discharge,the difference was statistically significant (t =6.845,t =7.231,P <0.05).No significant difference in the re -admission rate (16.67%,6.67%)was found between the two groups (χ2 =0.387,P >0.05).The complication rate in the intervention group was significantly lower than in the control group at 3th month after discharge(χ2 =5.124,P <0.05).Conclusion Discharge planning and home care can effectively relieve depression and anxiety in patients with peritoneal dialysis and improve the abilities of daily life.It is worth promoting and applying in clinic.

Maize seedlings were cultured in nickel or cadmium contaminated sand treated with α-naphthylacetic acid (NAA). The effects of NAA on nickel and cadmium uptake in roots, shoots, and subcellular fractions (cell wall, nuclei and remained parts of seedling cells) were determined. The data showed growth promotion when NAA was applied at low concentrations and inhibition at high concentrations. Uptake of nickel and cadmium content increased concurrently in roots and shoots. In the subcellular fraction, nickel and cadmium was greatest in the cell wall. The changes in growth had greatest correlation with nickel and cadmium content in the subcellular fraction.

Objective To observe the characteristics of cognitive impairment in patients with primary hypothyroidism.Methods A total of 90 primary hypothyroidism patients untreated with thyroid hormone were selected.All the 90 patients were divided into subclinical hypothyroidism group,mild or moderate hypothyroidism group and serious hypothyroidism group,and 30 patients in each group.The other 30 healthy volunteers were selected as controls.Mini-Mental State Examination (MMSE) and Montreal Cognitive Assessment (MoCA) were used for the testing their orientation,immediate memory,attention and calculation,delayed recalling,linguistic competence,visual space and execution,naming ability and abstracting power.One-way analysis of variance was performed to determine significant differences among four groups.Results The MMSE scores in subclinical hypothyroidism group,mild or moderate hypothyroidism group and serious hypothyroidism group((27.53 ± 2.16),(26.90±1.88) and (24.80 ± 2.10) respectively) were lower than those of the control (28.23 ± 1.33).The MoCA scores of the above hypothyroidism groups ((23.57 ± 3.33),(2 1.60 ± 2.81) and (20.53 ± 3.03) respectively) were also lower than that of the control (26.63 ± 2.31) (P ＜ 0.05).Except for orientation and immediate memory,statistical significances of the other cognitive function were existed between hypothyroidism groups and the healthy controls (P＜ 0.05).With the increase in severity of hypothyroidism,the abnormality of attention,calculation,linguistic competence,visual space and executive ability,naming ability and abstracting power were appearing gradually in hypothyroidism groups (P ＜ 0.05),and the scores were low(P＜ 0.05).Conclusion Defects of attention and calculation,delayed recalling,linguistic competence,visual space and execution,naming ability and abstracting power are existed in primary hypothyroidism patients.

The prescription of antibiotics for children with upper respiratory infections (URIs) in outpatient department of our hospital from November to December 2010 and from July to August 2011 were retrospectively analyzed.In 37 211 patient visits (prescriptions) with suppurative tonsillitis,laryngitis and other URIs,the antibiotic prescription rates were 90.45％ (3 626/4 009),78.77％ (2 642/3 354)and 52.25％ (15 596/29 848),respectively;the intravenous antibiotics rates were 78.22％ (3 136/4 009),60.58％ (2 032/3 354) and 16.47％ (4 915/29 848),respectively for above three conditions.For suppurative tonsillitis,the 3rd generation cephalosporins were the most commonly prescribed antibiotics (orally:51.07％,715/1 400) and(intravenously:54.88％,1 721/3 136),followed by the 2nd and first generation cephalosporins and macrolides.For laryngitis and other URIs,the 2nd generation cephalosporins were most frequently prescribed(orally:48.78％,538/1 103 and 39.16％,4 433/11 320,respectively) and (intravenously:54.38％,1 105/2 032 and 42.75％,2 101/4 915,respectively),followed by macrolides,3rd generation cephalosporins and first generation cephalosporins.No intravenous penicillin were prescribed.Oral penicillin were prescribed for 0-0.22％ of these children.The results indicate that antibiotics prescription rates are high for children with upper respiratory infections,particularly the prescription of broad spectrum antibiotics and intravenous antibiotics,while penicillin is seldom prescribed.

Objective To investigate the current treatment of childhood bronchiolitis and wheezy bronchitis in the outpatient clinic.Methods The medication treatments of the children under 2 years old with bronchiolitis were retrospectively analyzed in the outpatient department of the Children's Hospital Affiliated to the Capital Institute of Pediatrics,during the 2 periods from November to December 2010 and July to August 2011.These children were divided into 3 groups according to age,1-6 months old group,＞ 6 months to 1 year old group and ＞ 1-2 year old group.The medications prescribed,including antibiotics,steroid,β2 agonists and anticholinergic drugs were analyzed.Results Altogether 6 198 patient visits were included.Among them,4 764 (76.86％) visits received antibiotics,4 078 (65.80％) visits received intravenous antibiotics,3 018 (48.69％) visits received systemic glucocorticoids,2 980 (48.08％) visits received intravenous steroid,3 400 (54.86％) visits received nebulization treatment,3 381 (54.55％) visits received nebulized steroid,3 417 (55.13％) visits received inhaled bronchodilators,among them,3 384 (99.03％) visits received combined nebulization of β2 agonists and anticholinergic drugs;and 826 (13.33％)visits received oral bronchodilators.In the 3 age groups,the older the age group the higher the oral bronchodilators prescription rate (6.77％,11.50％ and 18.77％,respectively).While the prescription rate of intravenous steroid,inhaled steroid and inhaled bronchodilators were lowest in the oldest(＞ 1-2 year old) group (43.65％,49.64％ and 50.00％,respectively).The average cost was 166.08 Yuan per visit,and 69.06％ was spend on drug cost.Conclusions There is a wide spread use of antibiotics,glucosteroids and bronchodilators in the treatment of bronchiolitis and wheezy bronchitis.There is a need for national guideline of bronchiolitis to standardize the management of childhood bronchiolitis in outpatient clinic.

Objective To observe the inhibitory effect of atorvastatin on bleomycin-induced pulmonary fibrosis in SD rats and study their possible mechanism.Methods 30 male SD mice under SPF condition with average body weight of 250g were randomly allocated to three groups (n =10,each) of saline control group (control group),bleomycin-induced pulmonary fibrosis group (pulmonary fibrosis group) and atorvastatin treatment group (treatment group).Bleomycin (5mg/kg) (versus 0.2 ml saline in control group) were endotracheally instilled in pulmonary fibrosis group and the treatment group in order to establish the model of pulmonary fibrosis.Subsequently,the rats in the treatment group received daily atorvastatin (10 mg/kg) orally.5 rats in each group were sacrificed on 7th and 28th day after intratracheal instillation.Their lung tissues were taken and tested.The histological changes in the lungs were evaluated by hematoxylin-eosin and masson stain.The tumor necrosis factor (TNF-α) level and hydroxyproline content in lung tissues were measured by enzymelinked immunosorbent assay (ELISA).The expressions of Kruppel-like factor 2 (KLF2) protein and mRNA in lung tissues were measured by Western blotting and Real-Time PCR.Results The lung tissue in model group had significant bleeding and oozing inflammatory response on the 7th day and pulmonary fibrosis on the 28th day.Bleeding and oozing inflammatory response and pulmonary fibrosis were subdued in treatment group on the same days as compared to the model group.Hydroxyproline and TNF-α contents in lung tissue were significantly higher in model group than in control group (both P＜0.05).KLF2 protein and KLF2-mRNA expressions in lung tissues were significantly lower in model group than in control group (both P＜0.05).The above changes were partially reversed in treatment group.Compared to model group,treatment group showed that hydroxyproline and TNF-α contents in lung tissues were significantly reduced (both P＜0.05) and KLF2 protein and KLF2 mRNA expressions in lung tissues were significantly increased (both P＜ 0.05).Conclusions Atorvastatin can reduce the secretion of TNF-α and alleviate bleomycin-induced pulmonary fibrosis.The mechanism inhibiting fibrosis might be associated with up-regulation of KLF2-mRNA expression.

Objective This study was to investigate the association of certain single nucleotide polymorphisms (SNPs) of the ER with genetic susceptibility to ISS .Methods Genomic DNA was extracted from peripheral blood of children with ISS (n=355) and of normal growth and development individuals as controls from Jiangxi province (n=345) .The association between the ER gene polymorphisms with the height-related clinical traits was analysed .Results The allele T of rs6557177 is significantly lower in the ISS group than in the control group(P=0 .021 ,OR=0 .624) .The clinical indexes of two kinds of different genotypes were analyzed and compared ,there was no statistical difference(P>0 .05) .Conclusion The allele T of rs6557177 of ER gene is a protection factor of ISS .C gene and T gene grouping clinical parameters (hight ,weight ,IGF-1 ,IGFBP3 ,E2 and BMI) were analyzed and found no statistically significant difference .

Objective To evaluate the diagnostic value of shear wave elastography on deep venous thrombosis in different stage.Methods Models of rabbits'deep venous thrombosis have been established to noninvasively measure Young's moduli of thrombosis in day 1 ,4,7,10 and 14 respectively by shear wave elastography.Results The elastic modulus of thrombosis increased with increment of the stages of thrombosis.Specifically,the mean Young's moduli of thrombosis increased from (4.0±1 .42)kPa (day 1)to (25.71 ±6.09)kPa (day 14),with statistical differences between each two observation time points (P <0.05).Conclusions Shear wave elastography can effectively reflect the elasticity of thrombosis in different stages,and the stage of thrombosis can be estimated judging by quantitative ultrasound elasticity preliminarily.