Objective: To clone the Fab gene of a monoclonal antibody (mAb) BDI against human bladder cancer and its expression in E. coli. Methods: Fd and K genes of mAb BDI were cloned by RT-PCR and inserted into an Fab expression vector. Phage displaying Fab and soluble Fab were expressed in E. coli. The N-terminal sequence of VH region was corrected by PCR mediated mutagenesis. The antigen-binding characteristics of the Fab were tested by ELISA and immu-nohistochemistry. Results: Fd and K genes were cloned into the expressing vector p3MH and the phage displaying antibody and soluble Fab were expressed in E. coli, which showed weak binding activity to bladder cancer cells. Correction of the N-terminal sequence of the VHimproved the biding activity dramatically. The feasibility of the application of the Fab in phage antibody library screening was confirmed by a simulated panning procedure. Conclusion: The Fab gene of an anti-human bladder cancer mAb was expressed in E. coli. The importance of the N-terminal sequence on antibody binding activity was suggested.

Objective To investigate the changes of superoxide dismutase (SOD),malondialdehyde (MDA),hypersensitive c-reactive protein (hsCRP)in the process of age changes,in longevity areas in China,and the relationship between the above three indicators and hypertension and diabetes.Methods Residents who participated in the project of biomedical research of aging population conducted in 2014 were selected from 8 longevity Areas in China.2 467 people aged 40 and over attended the study,including 22 aged 40 and over,249 aged 60 and over,521 aged 70 and over,679 aged 80 and over,548 aged 90 and over,448 aged 100 and over.Information,including socio-demographic,were collected by self-designed questionnaire.Calibra-ted instruments were used by the clinical personnel who had worked over 3 years to manually check subjects’heath.The
fasting blood samples were collected by vacuum tube at early morning.The contents of plasma SOD,MDA,hsCRP and glu-cose were detected and compared among these elderly who were classified into different genders and different age groups.At the same time,compare the above three indicators of different subjects of hypertension,diabetes prevalence.Results In the groups aged 40 and over,60 and over,70 and over,80 and over,90 and over,100 and over,SOD were 55.55±8.82,54.49± 6.74,55.47±8.42,56.40±8.37,57.45±8.71 and 57.38±7.72 U/ml respectively in males,the difference between groups showed statistically significant (F =3.398,P <0.01)and 59.36± 10.98,56.08±7.98,57.41 ±8.54,57.38±8.18,59.16 ±7.71 and 57.90±8.66 U/ml respectively in females,the difference between groups showed statistically significant (F =2.667,P <0.01);MDA were 6.92±4.02,6.10±4.35,5.66±3.83,5.83±3.66,5.39±2.68 and 4.80±2.34 μmol/L re-spectively in males,the difference between groups were not statistically significant (F =2.198,P =0.052)and 8.93±8.20, 7.23±4.57,6.47±3.67,6.42±4.12,5.59±2.88 and 5.85±3.73 μmol/L respectively in females,the difference between groups showed statistically significant (F = 4.434,P < 0.01);positive rates of hsCRP were 11.8% (2/17),14.8% (26/176),19.3%(62/322),23.5%(77/327),26.1%(57/218)and 43.4%(33/76)respectively in males,the difference between groups showed statistically significant (χ2 =29.914,P <0.01)and 20.0%(1/5),16.4%(12/73),15.6%(31/199),20.7%(73/352),22.7%(75/330)and 28.0%(104/372)respectively in females,the difference between groups showed statistically significant (χ2 =14.078,P <0.01).In the groups of hypertension complicated with diabetes mellitus,hypertension,diabetes mellitus,non-diabetes hypertension SOD were 55.65±8.73,55.35±7.82,57.32±8.45 and 57.24±8.01 U/ml respective-ly,the difference between groups showed statistically significant (F = 3.694,P =0.011).MDA were 6.67 ±4.79,5.16 ± 2.70,6.01±3.87 and 5.66±3.15 μmol/L respectively,the difference between groups showed statistically significant (F =5.594,P =0.001);positive rates of hsCRP were 28.9%(54/187),29.5%(28/95),21.4%(275/1 288)and 21.2%(181/853)respectively the difference between groups showed statistically significant (F=8.749,P =0.033).Conclusion The re-sults indicated that the body had the ability to keep oxidation-antioxidant function stable in the process of aging.However, with the increase of age,the risk of cardiovascular disease was significantly increased,and it was more common for the aged suffering from inflammation.SODS,MDA,hsCRP are closely related to the occurrence of diabetes,high blood pressure,they were the reliable clinical indicators of oxidative stress degree.

Objective To evaluate the mechanical characteristics of systolic left ventricular(LV) transmural torsion in different LV electro-mechanical patterns using speckle tracking imaging. Methods Five open-chest canine models were employed for the acquirement of the basal, apical short-axis and four-chamber views of LV during baseline(BASE) and right atrial appendage(RAA), right ventricular apical (RVA), left ventricular lateral wall (LVL) and left ventrieular apical (LVA) pacing. Subendocardial (subend),subepicardial(subepi) and bulk rotation angle(RA) and segmental angle excursion(AE) at basal and apical level were analyzed using a dedicated workstation. LV torsion at different layers and bulk and global LV ejection fraction (EF) were calculated. Results ① There were no significant difference of transmural torsion and RA at basal and apical level between BASE and RAA pacing (P>0.05);② LV torsion of subend, subepi and bulk during RVA pacing were lower than those during RAA pacing(P0.05);LV torsion of subend and bulk during LVA pacing were lower than those during RAA pacing(P0.05);LV RA of subend,subepi and bulk at basal level during RVA and LVA pacing were lower than those during RAA pacing (P<0.05); ③ For normal electro-mechanical pattern, LV torsion of subend were significant higher than that of subepi(P<0.05), there only were a higher tendency for all pacing models (P>0.05); ④AE of segments near the pacing site decreased during different ventricle paeings (P<0.05); ⑤At BASE and during RAA pacing, LV bulk and subepi torsion were positively correlated to EF; RA of subend,subepi and bulk at basal level were positively correlated to EF. Conclusions LV transmual torsion are significantly depressed during RVA and LVA pacing. There is a spatial co-relationship between LV EF and torsion and rotation of bulk and subepi at basal level in normal LV electro-mechanical patterns.

To investigate the cell-killing effect and its possible mechanism of rClone30-hDR5 in combination with TRAIL on human hepatic carcinoma (HCC) cell line, first of all, recombinant plasmid pee12.4-hDR5 was introduced into HepG2 cells by liposome transfection. After five rounds of screening by flow cytometry, HepG2 cells expressing high levels of DR5 on cell surface were isolated. The cytotoxicity of TRAIL to selected cells was higher than that of TRAIL to HepG2 cells by MTT method (P < 0.01). The result suggested that the cloned hDR5 gene had biological activity. MTT assay showed that, rClone30- hDR5 in combination with TRAIL more efficiently inhibited the tumor growth of HepG2 cells compared to rClone30-hDR5 or TRAIL in vitro. The results of Annexin V-FITC/PI staining and Quantitative Real-time PCR indicated that rClone30-hDR5 in combination with TRAIL significantly increased the mRNA levels of caspase 3 and caspase 8, and induced the apoptosis of tumor cells. HepG2 cells were infected with rClone30-hDR5 or rClone30 at MOI of 1. The expression of hDR5 on tumor surface increased significantly by rClone30-hDR5 compared to that by rClone30, which contributed to the sensitivity to TRAIL. In conclusion, rClone30-hDR5 in combination with TRAIL has potential application value in cancer treatment.

Calcium phosphate cement (CPC) is considered as an important bone repairing materials due to its excellent biocompatibility, osteoconductivity and remodellability, the study about its performance is still a hot topic in the field of bone tissue engineering. Premixed calcium phosphate cements (PCPC) has advantages that can save operatiion time,be convenient to the operation and preservation compared with the traditional way of immediately mixing calcium phosphate cement. PCCP has overcome the shortcomings of uneven and inadequate mixing, and can be arbitrarily remodeled according to the shape of defect, thus researches on PCCP has also become more and more interested
Bone Cements ; chemistry ; Calcium Phosphates ; chemistry ; Humans ; Materials Testing ; Solubility

Poly(ADP-ribose)polymerase-1 (PARP-1) plays a significant role in the DNA repair process by catalyzing the transfer of ADP-ribose from NAD+ to its receptors. It is a promising anticancer drug target and many PARP-1 inhibitors have been developed and used in the clinical trial. In this work, a series of 3-(2-oxo-2-substituted acetamido)benzamides have been synthesized and their inhibitory activities against PARP-1 were evaluated. Of all the tested compounds, six compounds displayed inhibitory activities with IC50 values ranging from 0.23 to 5.78 µmol.L-1 . The binding pose of compound 5a was predicted using molecular docking to facilitate further structural modification.
Antineoplastic Agents ; Benzamides ; chemistry ; DNA Repair ; Drug Design ; Humans ; Molecular Docking Simulation ; Poly(ADP-ribose) Polymerase Inhibitors ; chemical synthesis ; chemistry ; Poly(ADP-ribose) Polymerases

Objective　To analyze the clinicopathological features of angiolymphoid hyperplasia with eosinophilia (ALHE). Methods　The pathological specimens of 7 cases of ALHE collected in our department from 1950 to 1999 were sectioned, stained and observed. Results　There were 3 pathological characteristics in ALHE: ①massive hyperplasia of capillaries in the dermis; ②the endothelial cells proliferated and swelled, projecting into vascular cavity like tombstones; ③mixed infiltration of lymphocytes and eosinocytes in the vessels. Conclusion　ALHE is a disease with local benign proliferated vessels, whose etiology and pathogenesis is still unknown. It is necessary to grasp the pathological changes of ALHE to distinguish it from other diseases.

Objective To investigate the effects of the intracellular delivery of HPV16E7_(49-57) epi- tope modified by cell-penetrating peptide HIV-Tat_(49-57) and its influential factors.Methods The unique HLA-A2+/H-2kb+ limited CTL epitope of HPV16E7 fused with a cell-penetrating sequence (HIV-Tat_(49-57)) was designed and a 18-mer peptide was synthesized with aid of polypeptide solid phase synthesis technique. The intracellular transport capabilities of these peptides were tested with indirect immunofluorescence assay and laser confocal microscopy.In addition,the CTL epitope and 18-met peptide were used to stimulate PBMC of healthy C57BL/6 mice,and the E7_(49-57) specific CTL responses were measured by LDH cytotoxicity detection kit in PBMC of those mice.Results The HIV-Tat_(49-57) peptide could efficiently assist HPV 16E7_(49-57) peptide in penetrating into BHK cells in a time and dose-dependent manner (P＜0.05).Further- more,the specific CTL activity induced by the 18-mer peptide was much stronger than that induced by the single epitope.Conclusion The results show that HIV-Tat_(49-57) can efficiently deliver the exogenous anti- genic peptide into the cytoplasm of live cells,and induce specific CTL response.This is a new way to de- sign peptide-based vaccine of HPV.

Objective To explore the feasibility of culturing dermal papillae cells (DPC) of hu- man hair in a serum-flee medium,and to observe the growth characteristics of these cells.Methods Cell culture flasks (plates) were pretreated with fibronectin,and DPC (2nd passage) were incubated with Williams E serum-flee medium supplemented with insulin-transferrin-selenite (ITS).Cells were observed by an inverted phase-contrast microscope.Proliferation of DPC was evaluated with 3-(4,5-dimethylthia- zol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and by their growth curve.Results In a serum-free medium,2nd passage DPC adhered to the flask surface within two to four hours of incubation; two to three days later,confluence,of the cells was observed,without noticeable proliferation.Four days later,cell connection was interrupted,isolated cells or cell clusters were seen,and detachment of some cells from the flask surface was observed.One to two weeks later,most cells had died.After incubation with 4% bovine serum for ten hours,cell proliferation was observed surrounding the remaining viable cell colonies. DPC growth curve showed stagnant phase and slow growth phase;however,log growth phase was not ob- served.Conclusion DPC could be successfully cultured in serum-free medium.However,the culture con- dition needs to be further optimized.

0.05).At the late stage (8-48 h) of incubation,the presence time of E7_(49-57)/K~b was significantly pro- longed on the surface of Tat-E7_(49-57)-incubated cells than that on the surface of other peptides-incubated cells (all P