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.

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

Objeetive To evaluate the changes of left ventricular(LV) global and segmental volume, LV outlet pressure and their co-relationship, and to access LV global and segmental systolic function and mechanical asynchrony in different LV electro-mechanical patterns using full volume three-dimensional echocardiography(3DE). Methods Nineteen open-chest canine models were employed for the acquirement of LV full volume dynamic 3DE imaging during right atrial appendage (RAA), right ventricular apical (RVA), LV lateral wall(LVL) and LV apical(LVA). LV outlet end-systolic pressure(ESP) was recorded simultaneously. End-systolic volume (ESV), end-diastolic volume (EDV), global and segmental ejection fraction(EF) and systolic dyssynchrony index(SDI) of LV were measured and calculated using a dedicated workstation. The average ascending rate of LV pressure during systole(+ dp/dt) and the average descending systolic pressure(ESP), + dp/dt and - dp/dt during RVA pacing were lower than those during RAA pacing (P <0.05). SDI during RVA pacing was higher than that during RAA pacing(P<0.05). ESP, + dp/dtand - dp/dt during LVL and LVA pacing were lower than those during RAA pacing (P <0.05). There and LVA pacing was higher than that during RVA pacing (P <0.05),SD1 during LVL pacing was lower than that during RVA pacing (P <0.05), there was no significant difference of SDI between RVA and LVA and LVL pacing. Segmental EF of septum and apex during LVI. pacing were higher than those during LVA pacing (P <0.05). @Segmental EF of anterior and post septum and all apical segments (except lateral wall) during RVA pacing were lower than those during RAA pacing (P <0.05). Segmental EF of lateral and anterior wall during I.VI. pacing were lower than those during RAA pacing (P <0.05). Segmental EF of anterior wall and anterior septum during LVA pacing were lower than those during RAA parameters. Conclusions The global and minority segmental systolic function of LV during RAA pacing could be reduced compared with normal sinus rhythm. All the ventricular pacing worsen LV systolic and diastolic function compared with RAA pacing. LV systolic function during LVL pacing was superior to RVA pacing. During ventricular pacing,the systolic function at nearby segments of the pacing site was depressed.

BACKGROUND: Adequate preparation of donors and recipients prior to living-related donor renal transplantation, short warm and cold ischemia time for donor kidney, good histocompatibility of human leukocyte antigen match, and low postoperative rejection incidence provide feasibility for use of low-dose immunosuppressive agents after living-related donor renal transplantation. OBJECTIVE: To investigate the safety and effectiveness of low-dose calcineurin inhibitors (CNI), an immunosuppressive agent, in living-related donor renal transplantation. METHODS: A total of 38 recipients who underwent living-related donor renal transplantation at the Center of Renal Transplantation of the First Affiliated Hospital of Nanjing Medical University from January 2006 to June 2008 were randomized for treatment with mycophenolate mofetil (750 mg twice a day), prednisone, and either standard-dose CNI (n=18) or low-dose CNI (n=20) during 12 months post-transplantation. Ciclosporin A was given orally (starting dose, 6 and 4 mg/kg per day, respectively) in two divided doses to achieve the 12-hour whole blood concentration as measured by fluorescence polarization immunoassay. The starting dose of tacrolimus was 0.12 and 0.08 mg/kg per day respectively, and its whole blood concentration was measured by enzyme-multiplied immunoassay technique. After transplantation, patients were followed up. Renal function, pulmonary infection, liver dysfunction, and CNI nephrotoxicity at different time periods were compared between different regimens. RESULTS AND CONCLUSION: During 12 months post-transplantation, patient death occurred in one of 18 patients (5.6%) in the CNI standard-dose group and none of 20 patients (0%) in the CNI low-dose group. There was no significant difference in renal function and acute rejection between CNI standard-dose and CNI low-dose groups (P > 0.05). The incidence of liver dysfunction and CNI nephrotoxicity was significantly lower in the CNI low-dose group than in the CNI standard-dose group (P < 0.05). In addition, a low-dose CNI regimen helped recipients to lessen the economic burdens. These findings indicate that it is effective, safe and economical to use a low-dose CNI regimen in living-related donor renal transplantation.

Human ScFv against botulinum neurotoxin serotype A(BoNTa) was modified by fusing human IgG1 Fc to C terminal of ScFv. ScFv-Fc fusion protein was expressed at high level over 30% of total host cell proteins in E.coli. Recombinant protein existed in inclusion body form. Renatured ScFv-Fc was purified to 90%~95% by Protein G Sepharose column. In vitro ScFv-Fc could bind specific to toxiod BoNTa in ELISA. Recombinant ScFv-Fc had similar relative affinity to parent ScFv and had improved stability.

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

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.