Objective To construct a human phage single chain-antibody library, and to sieve out the antibody ScFv against lung cancer from the library. Methods Total RNA was abstracted from lymph node tissue of the lung cancer, and was used to amplify V_H and V_L gene by RT-PCR. V_H and V_L were joined by a DNA linker by SOE-PCR to form the single chain variable fragment ( ScFv) gene. ScFv gene was coloned into the phage vector pCANT-AB5E. Panning against lung cancer cell line A549 was performed and positive clones were chosen for soluble expression. Results A recombination phage single chain-antibody library was constructed. After 4 rounds panning, the number of eluted phages increased by 115 times. Positive reactions to A549 were detected in 7 of 10 random clones. The human ScFvs against lung cancer were produced and confirmed by SDS-PAGE and ELISA analysis. Conclusion ScFvs against lung cancer were acquired by the construction of phage single chain-antibody library. The soluble ScFvs has specificall avidity to human lung cancer cells.

Aim To screen and identify anti-CCR7 single chain fragments variable(scFv)from lager phage library and to detect the scFv efficiency.Methods The insert ratio of ScFv antibodies library was identified by PCR.The products digested by Sfi I/Not I double enzyme.Panning against breast cancer cell and CCR7 were performed four and three rounds respectively.Positive clones were transformed to E.coli HB2151, and their dissolvability was assayed.The soluble scFv was purified by affinity chromatography, and its relative molecular mass was determined by Western blot.The ELISA assay was used to identify the immunocompetence of the antibody.Immunocytochemical staining and radioimmunoimaging were employed to determine the affinities of scFv binding with CCR7 in cell line and in nude mice.Results The insert ratio of ScFv gene was 90%(18/20), enzyme digest reaction showed the aim products on 1% agarose gel.ScFvs were obviously enriched after 7 round panning.Western blot result showed soluble scFv's molecular mass was about 34 KD.ELISA analysis showed dissolved antibody had high immunocompetence to MDA-MB-435 s cells.Immunocytochemical staining and radioimmunoimaging indicated that ScFvs were bound efficiently to MDA-MB-435 s cells which expressed CCR7.Conclusions ScFvs against CCR7 are successfully acquired by screening the phage antibody library.The soluble ScFvs have high affinity and specifical binding to human breast cancer cells.

Objective To construct a human phage single chain-antibody library,and to sieve out the antibody ScFv against lung cancer from the library.Methods Total RNA was abstracted from lymph node tissue of the lung cancer,and was used to amplify VH and VL gene by RT-PCR. VH and VL were joined by a DNA linker by SOE-PCR to form the single chain variable fragment (ScFv) gene. ScFv gene was coloned into the phage vector pCANTAB5E. Panning against lung cancer cell line A549 was performed and positive clones were chosen for soluble expression.Results A recombination phage single chain-antibody library was constructed. After 4 rounds panning,the number of eluted phages increased by 115 times. Positive reactions to A549 were detected in 7 of 10 random clones. The human ScFvs against lung cancer were produced and confirmed by SDS-PAGE and ELISA analysis.Conclusion ScFvs against lung cancer were acquired by the construction of phage single chain-antibody library. The soluble ScFvs has specificall avidity to human lung cancer cells.

Objective To assess the evaluation of SRT (Stereotactic radiotherapy ) and 3D-CRT (Three-dimensional conformal ra-diotherapy)on treatment of brain metastasis from lung cancer .Methods From June 2009 to June 2011 ,74 patients with multiple brain metastasis(brain metastasis ≤3 ,tumor mass ≤3 cm) from lung cancer were analyzed retrospectively ,37 patients received 3D-CRT alone were retrospectively compared with 37 patients who received SRT alone .the results was evaluated by median survival time(MS) ,local control(LC) and toxicity effect .Results The median survival time(MS) was 9 .3 ,which 8 .6 months after 3D-CRT ,and 10 .6 months after SRT .the local control rate was 73 .5% for 3D-CRT ,and 79 .6% for SRT after 3 months .there was no difference between two groups on toxicity effect from stastic .Conclusion The SRT was a efficacious methods for the treatment of brain metastasis from lung cancer ,which could improve the local control rate ,but there was no more toxiticy .

Objective To investigate the feasibility of magnetic resonance diffusion tensor imaging (DTI) as a noninvasive way for assessment of kidney transplant function in the early posttransplantation period.Method Fifty-one kidney transplant recipients less than 1 month after kidney transplantation and 26 age-matched healthy volunteers were included and examined using a fatsaturated echo-planar DTI sequence in oblique-coronal orientation at 3.0 Tesla magnetic resonance (MR) imager (diffusion directions=6,b =0,300 s/mm2).According to the estimated glomerular filtration rate (eGFR) recorded and calculated on the examined day,all subjects were divided into four groups:group 1,healthy volunteers (n =26) ; group 2,eGFR≥60 mL/min/1.73 m2 (n =24) ; group 3,30≤eGFR＜60 mL/min/1.73 m2 (n =19) ; group 4,eGFR＜30 mL/min/1.73 m2 (n =8).Mean apparent diffusion coefficient (ADC) and mean fractional anisotropy (FA) were determined separately for the cortex and the medulla.The paired Students t test was used to compare ADC and FA between cortex and medulla within each group.ADC and FA between groups were compared by using the one-way analysis of variance test.Relationship between ADC and FA with eGFR of the transplants was assessed by using Pearson correlation analysis.Result Mean cortical FA was significantly higher in group 1 than in other three groups (P＜0.01 for all),while differences among groups of allograft recipients were not significant (P＞0.05 for all).There was a gradually decreasing trend of medullary FA and ADC,and cortical ADC from group 2 to group 4,and the differences among groups were all pronounced (P＜0.05 for all).In renal allografts,there was a significant positive correlation between eGFR and medullary FA,medullary ADC as well as cortical ADC (r =0.812,0.756,0.757,respectively,P＜0.01).The cortical-medullary discrimination of FA-map and ADC-map in group 3 and group 4 apparently decreased.Meanwhile,DTI revealed that the radial diffusion tracts in the medulla of group 3 and group 4 obviously broke off and reduced,especially in group 4.Conclusion DTI is a promising way to evaluate kidney transplant function early after transplantion,and can quantitatively and visually distinguish transplants with different functions.

Objective To investigate the relationship between frailty and serum biomarkers in the elderly. Methods A total of 371 elderly individuals aged 60 years and above with complete medical data were recruited during health examinations. Frailty phenotype assessment and comprehensive geriatric assessment were conducted.Serum levels of interleukin-6 (IL-6 ) ,high sensitivity C-reactive protein(hs-CRP) ,tumor necrosis factor-α(TNF-α) ,homocysteine(Hcy) ,insulin-like growth factor-1(IGF-1) ,25-hydroxyvitamin D[25(OH)D] ,folic acid and vitamin B12(VitB12) were detected by enzyme-linked immunosorbent assays ( ELISA ) and chemiluminescence immunoassays. Associations between frailty and the above factors were analyzed. Results Serum levels of IL-6 ,TNF-α ,Hcy and IGF-1 were significantly elevated along with progressive increase in frailty severity(all P＜0.05).There were a downward trend in serum 25(OH)D levels and an upward trend in serum hs-CRP ,folic acid and VitB12 levels as frailty severity increased ,but the changes did not amount to any statistical significance(all P＞0.05).Logistic regression analysis showed that ,after adjusting for age ,gender ,body mass index (BMI)and some clinical aspects (hearing loss ,urinary incontinence ,pain ,malnutrition ,cognitive dysfunction ,decreased activities of daily living ,depression , insomnia ,and anemia) ,serum levels of IL-6(OR=1.012 ,95% CI=1.005-2.041 ,P=0.033) ,IGF-1 (OR= 1.017 ,95% CI = 1.011-1.118 ,P= 0.021)and Hcy (OR= 1.007 ,95% CI :1.002-1.073 ,P=0.047)were significantly associated with frailty status. Conclusions Serum levels of IL-6 ,Hcy and IGF-1 are related to frailty status and may be used as potential biomarkers for the assessment of frailty in older adults.