Objective To isolate specific humanized anti-D-dimer scFv(single chain Fv) antibody from scFv phage libraries. Methods Isolate anti-D-dimer positive clones from Tomlinson I + J phage libraries by three rounds of panuing, then sequence monoclonal genes by bideoxy-mediated chain termination and express soluble scFv antibody; Pick out anti-D-dimer antibodies with high specificity and affinity by ELISA.Results After three rounds of selection from human scFv phage libraries Tomlinson I and J, 38 monclonal specific anti-D-dimer scFv fragments were selected. By polyclonal and monoclonal phage ELISA and gene sequencing, 20 different full-length monoclonal scFv phages were identified, the result of soluble scFv ELISA showed that 20 full-length monoclonal scFv were expressed smoothly. According to the result of soluble scFv ELISA, in 5 scFv antibodies with high value of A450 selected, 3 scFv antibody fragments showed high specific and affinity. Conclusion Antibody phage display was an effective, rapid method to isolate anti-D-dimer antibodies with high specificity and affinity.

Aspergillosis ; etiology ; therapy ; Aspergillus ; Humans ; Lung Diseases, Fungal ; etiology ; therapy ; Male ; Middle Aged ; Paraquat ; poisoning ; Treatment Outcome

Objective To investigate the effect of microRNA-1 (miR-1) on the fibrosis of cardiac fibroblasts induced by high glucose. Methods The primary cultured fibroblasts from 1-3 days old Sprague-Dawley (SD) rats, were randomly divided into 4 groups (n = 3): normal glucose + lentivector-vehicle (CON+Lv-Vehicle group), normal glucose + lentivector-miR-1 (CON+Lv-miR1 group), high glucose + lentivector-vehicle (HG+Lv-Vehicle group), high glucose + lentivector-miR-1 (HG+Lv-miR1 group). Fibroblasts were cultured in glucose concentration 5.5 mmol/L and 25 mmol/L DMEM culture, and were injected lentiviral vector carrying miR-1 silencer sequence or the same volume of lentiviral vector. After 12 hours, the medium was replaced with fresh complete medium. After 3 days when transfection efficiency was up to 90%, the cellular miR-1 content was detected by real-time reverse transcription-polymerase chain reaction (qRT-PCR). The secretion of collagen Ⅰ and Ⅲ were measured by enzyme linked immunosorbent assay (ELISA). Expression of collagen Ⅰ and Ⅲ, matrix metalloproteinase 2 and 9 (MMP-2, MMP-9), autophagy related protein LC3B-Ⅱ, P62/SQSTM1 and Cathepsin D were assessed by Western Blot. Results Compared with the CON+Lv-Vehicle group, the content of miRNA in the CON+Lv-miR1 group had no statistical significance. Compared with the CON+Lv-Vehicle group, high glucose increased the amount of miR-1 (2-ΔΔCt: 1.82±0.17 vs. 1.00±0.04), collagen Ⅰ and Ⅲ secretion (mg/L: 14.55±0.33 vs. 7.28±0.22, 157.50±13.22 vs. 61.25±8.54) and expression (gray value: 432.35±56.00 vs. 100.00±15.00, 320.35±47.00 vs. 100.00±15.00), the level of MMP-2, MMP-9 and the expression of autophagy related protein LC3B-Ⅱ and P62/SQSTM1 (gray value: 249.0±21.0 vs. 100.0±15.0, 142.3±20.0 vs. 100.0±16.0, 178±19 vs. 100±14, 378.3±20.0 vs. 100.0±15.0), decreased the expression of lysosomal associated protein Cathepsin D (gray value: 60±14 vs. 100±10), and the differences were statistically significant (all P < 0.01). Compared with the HG+Lv-Vehicle group, the amount of miR-1 in the HG+Lv-miR1 group was significantly decreased (2-ΔΔCt: 1.21±0.10 vs. 1.82±0.17), collagen Ⅰ and Ⅲ secretion (mg/L: 10.68±0.54 vs. 14.55±0.33, 87.25±13.55 vs. 157.50±13.22) and expression (gray value: 179.41±45.00 vs. 432.35±56.00, 173.41±50.00 vs. 320.35±47.00), the level of MMP-2, MMP-9 and the expression of autophagy related protein LC3B-Ⅱ and P62/SQSTM1 (gray value: 172.0±23.0 vs. 249.0±21.0, 90.0±17.0 vs. 142.3±20.0, 138±15 vs. 178±19, 265.0±17.0 vs. 378.3±20.0) in the HG+Lv-miR1 group were decreased and the expression of lysosomal associated protein Cathepsin D was higher (gray value: 110±17 vs. 60±14), and the differences were statistically significant (all P < 0.05). Conclusions The expression of miRNA-1 was up-regulated in cardiac fibroblasts cultured in high glucose, and miRNA-1 silencing inhibited cardiac fibroblast induced fibrosis in high glucose. The mechanism may be related to the recovery of autophagy flux, up-regulation of Cathepsin D expression and inhibition of collagen production.

Twelve populations of Inula lineariifolia were used as materials to measure morphological characteristics, photosynthetic parameters and chemical constituents. It aims to provide a theoretical basis for germplasm resources evaluation. The results showed that I. lineariifolia had relatively rich morphological diversity, there were significant differences of morphological characteristics, photosynthetic parameters and chemical constituents among populations. There was positive correlation on morphological characteristics and P(n). Twelve populations were divided into three-type. The three populations of Xuyi, Mingguang and Fengyang were of narrower-longer leaf, bigger biomass,better photosynthetic and higher chemical constituents. Then they were classified for a similar group. It proved that the three populations were more suitable for cultivation and promotion.
Biomass ; China ; Flowers ; chemistry ; growth & development ; metabolism ; Inula ; chemistry ; classification ; growth & development ; metabolism ; Photosynthesis ; Plant Leaves ; chemistry ; growth & development ; metabolism ; Plant Stems ; chemistry ; growth & development ; metabolism

ObjectiveTo explore the procedures and methods for genetic diagnosis in one nonsyndromic variants of congenital neutropenia (NSVCN) patient and its pathogenic mutation.Methods Genomic DNA was prepared from one NSVCN patient who had progressed to chronic myelomonocytic leukemia and ELA2,HAX1,WASp and GFI1 genes were amplified and sequenced.Results A novel compound heterogeneous mutation consisting of two frame-shift mutations (c.430-1insG and c.655- 9del5bp) was found in HAX1 gene.ConclusionA practically genetic diagnosis procedure for NSVCN has been established,and the novel HAX1 gene mutation may contribute to the etiology of NSVCN.

Objective To assess the sleep quality of postoperative patients with breast cancer,and find out influencing factors.Methods The measurement was to adopt Athens Insomnia Scale,Hamilton Depression Scale and Big Five Personality Scale to do questionnaire for 60 breast cancer postoperative patients and 60 healthy subjects.Results 88.34％ patients had sleep difficulty after operation.The score of sleep quality of patients (12.60 ± 4.65) were higher than that in control group (3.70 ± 3.77,P ＜ 0.05).Similarly,according to the stepwise multiple regressions,the score of sleep quality of patients were positively associated with anxiety,adaptability of personality and negatively associated with openness of personality.Conclusion Sleep problems of breast cancer postoperative patients are quite severe.The influencing factors of sleep quality of patients are anxiety,openness and adaptability of personality.

Objective To investigate the clinical value of bile reinfusion combined with enteral nutrition before R0 resection of hilar cholangiocarcinoma.Methods The clinical data of 46 patients with hilar cholangiocarcinoma who underwent R0 resection at the Affiliated Drum Tower Hospital of Medical School of Nanjing University from January 2010 to January 2014 were analyzed retrospectively.The clinical data of 21 patients (the experimental group) who received bile reinfusion by preoperative percutaneous transhepatic cholangiography and drainage (PTCD) combined with enteral nutrition with nasojejunal tube were compared with 25 patients (the control group) who received neither PTCD nor enteral nutrition.The differences in the clinical indicators between the 2 groups were analyzed.Results In the experimental group,the levels of ALT,AKP,TBil and DBil decreased significantly after PTCD as compared with the original levels (t =4.433,4.547,5.648,4.681,P ＜ 0.05).The operation time and blood loss in the experimental group was reduced significantly when compared with the control group (t =-2.810,-2.047,P ＜ 0.05).The volume of postoperative albumin transfusion and the postoperative hospitalization in the experimental group was significantly reduced when compared with the control group (t =-3.083,t =-3.083,P ＜ 0.05).Conclusion Bile reinfusion combined with enteral nutrition facilitated the recovery of preoperative hepatic function in patients who underwent R0 resection of hilar cholangiocarcinoma,thus improving the safety of surgery and facilitated postoperative rehabilitation.

Objective To explore the relationship between job burnout and personality characteristics of prison police,and provide a basis for prevention and intervention of prison police burnout.Methods Using Maslach burnout inventory general survey(MBI-GS) and Catteii the sixteen factor questionnaire(16PF) to investigate 217 prison police in Beijing.Results The overall prevalence of burnout syndrome among prison police was 64.7％.The prevalence of emotional exhaustion(11.09±5.86),depersonalization(6.38±4.81) and diminished personal accomplishment(17.96± 2.07) was 46.5％,25.6％,33％ respectively.Compared with the national norm of 16PF,the 10 factors of warmth(t=-2.01,P=0.007),reasoning(t=9.22,P=0.000),dominance(t=6.46,P=0.000),liveliness(t=4.01,P=0.000),social boldness(t=-2.00,P=0.047),sensitivity(t=-7.80,P=0.000),abstractedness (t=-2.89,P=0.004),privateness (t=4.35,P=0.000),perfectionism(t=3.19,P=0.002) were significant differences.The emotional exhaustion was significantly related to emotional stability,dominance,liveliness,social boldness and tension.The depersonalization was significantly related to emotional stability,abstractedness and tension.The diminished personal accomplishment was significantly related to self-reliance.Conclusion Prison police burnout situation is more serious,and personality characteristics has a significant impact on job burnout.

OBJECTIVEThis research aimed to detect the expression levels of ffh gene in Streptococcus mutans (S. mutans) UA159 under different pH conditions, analyze the effect of pH on the expression of ffh gene in S. mutans, and identify the factors regulating the ffh gene expression.

METHODSSamples of S. mutans were collected at different growth stages (4 h, 18 h) and pH values (pH 4.0-7.0). Fluorescence quantitative real-time polymerase chain reaction (qRT-PCR) was used to measure the relative mRNA expression and trend of the target gene ffh in S. mutans at different growth stages and pH values.

RESULTSqRT-PCR results showed that the ffh gene expression decreased along with pH at 4 h, but the expression increased with decreasing pH at 18 h. Under the same pH conditions, the ffh gene expression was significantly different between 4 h and 18 h (P < 0.05).

CONCLUSIONGrowth stage and pH value influenced the ffh gene expression in S. mutans.

OBJECTIVE To study the content and distribution characteristics of thymic stromal lymphopoietin (TSLP) in nasal mucosa of murine model of allergic rhinitis.METHODS 6-week-old BALB/c mice were divided into control group and experimental group.Mice of experimental group were sensitized by means of dropping OVA in nasal cavity and intraperitoneal injection of OVA. Meanwhile,mice of control group were dealt with physiological saline instead of OVA.The content of TSLP in nasal mucosa of mice was tested by means of real-time PCR and its distribution was detected by means of immunohistochemistry staining.RESULTS The content of TSLP was obviously higher in nasal mucosa of experimental group than that of control group.It was distributed mainly in lamina propria and on the surface of epithelium.CONCLUSION TSLP may play an important role in allergic rhinitis of the mice.