The biological activity of ADCC by anti-CD20 monoclonal antibody was determined by BioGlo™ Luciferase Assay System using Jurkat/NFAT-luc+FcγRIIIa cell line as effector cell and WIL2-S cell line as target cell. The developed method was verified for specificity, precision and accuracy. Anti-CD20 monoclonal antibody showed a dose-response mode by the developed method, and the determination result complied with the following four-parameter equation: y = (A-D)/[1 + (X/C)(B)] + D. The optimized parameters of the method were determined including the antibodies diluted concentration (18,000 ng·mL(-1)), dilution rate (1:5), the ratio of effector cell and target cell (6:1), and induction time (6 h). The values of eight independent tests have passed a statistical test for curve regression analysis, linear or parallelism, which showed the method possessed good specificity. Four different dilute groups of recovery rates sample were determined for 3 times, and the result showed mean relative potencies of (44.39±3.93)%, (72.74±2.78)%, (128.28±7.01)% and (168.19±2.70)% respectively, with a variation coefficient of less than 10%, and the recoveries of (88.78±7.85)%, (96.99±3.70)%, (102.63±5.61)% and (112.12±1.80)% respectively. A novel reporter gene method for determination of biological activity of ADCC by anti-CD20 monoclonal antibody was successfully developed, which showed strong specificity, good reproducibility and high accuracy, and might be used routinely.
Antibodies, Monoclonal, Murine-Derived ; pharmacology ; Antibody-Dependent Cell Cytotoxicity ; Antigens, CD20 ; immunology ; Genes, Reporter ; Humans ; Reproducibility of Results ; Rituximab

OBJECTIVETo observe the effects of telmisartan on Kv1.3 and Kv1.5 potassium channels expressed in Xenopus oocytes.

METHODSKv1.3 and Kv1.5 potassium channel currents expressed in Xenopus oocytes were recorded and observed in the absence and presence of telmisartan using standard two-microelectrode voltage clamp techniques.

RESULTSTelmisartan resulted in a concentration- and voltage-dependent inhibition effect on Kv1.3 channel current (IC(50) 2.05 micromol/L)and on Kv1.5 channel current (IC(50) 2.37 micromol/L).

CONCLUSIONSTelmisartan blocks open-state Kv1.3 channel which could be one of the mechanisms related to its immunomodulatory and anti-atherosclerosis effect. Telmisartan also blocks open-state Kv1.5 channel which might partly account for its effect on reducing the incidence of atrial fibrillation.

Animals ; Benzimidazoles ; pharmacology ; Benzoates ; pharmacology ; In Vitro Techniques ; Kv1.3 Potassium Channel ; drug effects ; Kv1.5 Potassium Channel ; drug effects ; Oocytes ; drug effects ; metabolism ; Patch-Clamp Techniques ; Xenopus

Objective:To construct the Escherichia coli (E. coli)expression system for preparation of the bone morphogenetic protein-2 (BMP2)with collagen-binding domain (CBD),and to study the methods and conditions for expression, purification and renaturation of CBD-BMP2.Methods:CBD sequence was cloned into the N-terminal of BMP2 sequence, the recombinant vector pet21b/CBD-BMP2 was constructed and transformed into E.coli BL21.The expression of recombinant protein was induced using isopropylβ-D-thiogalactopyranoside (IPTG) at 37 ℃.Ni-NTA chelate chromatography was used to purify CBD-BMP-2.Denaturing CBD-BMP2 was refolded by dilution method using ultrapure water.The refolding CBD-BMP2 was filtered through a 0.22μm microfiltration membrane for degermation.The recovery rate was calculated by the ratio of the protein concentration before and after degermation. The expression, purification, and renaturation of recombinant protein were detected by SDS-PAGE method.The concentration of CBD-BMP2 was detected by BCA assay.Results:The recombinant vector pet21b/CBD-BMP2 was successfully transformed into E.coli BL21,and the recombinant protein was expressed as inclusion bodies in E.coli.The SDS-PAGE results showed denaturing protein was dissolved in supernatant of lysis buffer with 8 mol·L-1 urea and the purified recombinant protein existed in elution buffer B with relative molecular mass about 14 000.Two bands (14 000 and 28 000)were seen in the SDS-PAGE picture,which indicated that the monomer was successfully refolded into dimer by dilution method.The concentrations of recombinant protein before and after degermation were 110 and 80 mg · L-1 , respectively, and the recovery rate was about 73%. Conclusion:The recombinant vector pet21b/CBD-BMP2 is transformed into E.coli BL21 successfully,and the recombinant CBD-BMP2 is expressed and refolded efficiently. The methods of prokaryotic expression system for preparing recombinant CBD-BMP2 protein are established.

OBJECTIVETo evaluate the clinical effectiveness of transurethral seminal vesiculoscopy (TUSV) combined with finasteride in the treatment of recurrent hemospermia.

METHODSThis study included 32 patients with recurrent hematospermia, with the disease course of 3 months to 4 years. After administration of finasteride at 5 mg/d for 2 weeks, the patients underwent TUSV for both exploration of the causes and treatment, followed by medication with finasteride at the same dose for another 2 weeks. Postoperative follow-up was conducted for observation of the outcomes and complications.

RESULTSTUSV was successfully accomplished in all the 32 cases, which revealed 16 cases of seminal vesiculitis, 10 seminal calculi, 1 seminal vesicle cyst, 2 seminal vesicle polyps, and 3 seminal vesicle abscess. The operative time was 20 to 51 (31.0 +/- 5.2) minutes. Postoperative complications included 1 case of acute epididymitis and 3 cases of breast discomfort within the first 4 weeks. No incontinence, urethral stricture, rectal injury, retrograde ejaculation, and sexual dysfunction occurred postoperatively. All the patients but 1 were followed up for 6 months to 2 years. Twenty-nine of the cases were cured, and 2 experienced recurrence.

CONCLUSIONTransurethral seminal vesiculoscopy combined with finasteride is safe and effective for the treatment of recurrent hemospermia.

Adult ; Endoscopy ; methods ; Finasteride ; therapeutic use ; Follow-Up Studies ; Hemospermia ; therapy ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

Objective To describe the prevalence of deliberate self-harm (DSH) and suicidal behaviors (SIB) as well as the relationship between them in a college student population.Methods A total of 4063 medical students were selected under the cluster sampling method in Anhui province.Data were analyzed by Pearson Chi-square and logistic regression.Results A total of 4063 ( 13.4％ )students reported that they had deliberately harmed themselves during the past 12 months.The acts of DSH with 1 and more than or equal to 2 times occurrence accounted for 3.7％ and 9.6％ among all the respondents.The act of DSH was significantly higher among boys than that among girls.Rates of suicide ideation,suicide plan,attempted suicide and SIB in the last year were 4.5％,1.4％,0.6％ and 4.9％,respectively.However no statistically significant difference was found in different sex.Students in the DSH group that with SIB were found to have lower positive coping levels (OR=0.5,95％CI:0.3-0.8),getting less support from fricnds (3-5 vs.less than or equal to 2:OR=0.6,95％CI:0.3-0.9;more than or equal to 6 vs.less than or equal to 2:OR=0A,95％CI:0.2-0.8),with higher negative coping levels (OR=2.1,95％CI:1.2-3.7),having more serious depressive symptoms (OR=2.9,95％ CI:1.6-5.2) and anxiety symptoms (OR=2.2,95％ CI:1.2-3.8),having more serious sleeping problems (OR=1.7,95％CI:1.1-2.8 ) and perceived fat (fat vs.moderate:OR=2.0,95％CI:1.1-3.6) than the DSH group without SIB.The rates of SIB in students with DSH behavior were significantly higher than those in students without those behaviors (OR=4.7,95％ CI:3.5-6.4).Psyehosocial variables could attenuate the relationship between the DSH status and suicidal events (OR=3.3,95％CI:2.4-4.5).The DSH frequency exhibited a curvilinear relationship to SIB (OR=3.1-10.0) and psychosocial variables also attenuated this relationship (OR=2.4-5.9).Conclusion It was well known that SIB was not a suicidal gesture but our findings suggested that the presence of DSH might trigger the suicidal attempts.

Background About one third of congenital cataract is associated with inheriting factor.The inherited heterogeneity has been found in congenital cataract.To seek the pathogenic gene is essential for the gene therapy. Objective Present study was to map and identify the causal gene for autosomal dominant congenital cataract (ADCC) in a Chinese family. Methods The clinical features of all affected members in this family were examined.Blood samples were collected from eleven family members for genetic linkage analysis.Polymorphic microsatellite markers were selected from the regions which harbor all known loci linked with ADCC.Universal fluorescent-labeled M13 primer was used in linkage analysis.Direct genomic sequencing was used to evaluate the candidate gene for example CRYBB2 gene.This study followed Helsinki Declaration and was proved by Tianjin City Ethic Committee.Written informed consent was obtained from each SUbject before any medical procees. Results The maximum two-point LOD score of 1.20 was obtained for marker D22S315 (θ=0).The LOD score of 0.6 was obtained for marker D16S3068.No mutation in all exons of CRYBB2 gene was found in the family. Conclusion CRYBB2 gene associated with ADCC was excluded from the family.A genome-wide linkage screening should be conducted.Genotyping with microsatellite markers using Universal fluorescent-labeled M13 primer can decrease the cost and obtain the same result.

OBJECTIVETo quantitatively assess the effects of cardiac resynchronization therapy (CRT) in patients with advanced congestive heart failure by real-time 3-dimensional(3D) echocardiography (RT-3DE).

METHODSEighteen patients with advanced congestive heart failure underwent CRT with New York Heart association(NYHA) class III and IV and wide QRS complex (>120 ms) were included (17 dilated cardiomyopathy and 1 ischemic cardiomyopathy). Before CRT and 8 months after CRT, the clinical and RT-3DE parameters and outcome were analyzed.

RESULTSThe biventricular pacemaker was successfully implanted in 17 patients (94.4%). Compared with before CRT, NYHA class of patients decreased by 1.5 class (P < 0.01), left ventricular ejection fraction increased by 25% (P < 0.01), left ventricular end systolic volume decreased by 38% (P < 0.01), left ventricular systolic dyssynchrony index (SDI) improved significantly (14.2% before CRT vs. 9.8% after CRT, P < 0.01 ) post CRT. Change in SDI and change in LVEF was positively correlated (r = 0.62, P < 0.01) . The procedure complications and outcome during and after CRT included coronary sinus dissection (n = 1), left ventricular lead dislodgement (n = 1), phrenic nerve stimulation (n = 1), sudden cardiac death (n = 1). Three non-response patients were complicated with atrial fibrillation, nonspecific intraventricular block and dilated cardiomyopathy with postero-lateral scar tissue.

CONCLUSIONSCRT could improve the cardiac function, correct the mechanical desynchronization and reverse left ventricular remodeling in patients with congestive heart failure, and SDI quantification by RT-3DE could predict increase of LVEF after CRT, however, there were complications related to the implantation procedure and possibilities of non-response.

Adult ; Aged ; Cardiac Pacing, Artificial ; methods ; Cardiac Resynchronization Therapy ; Echocardiography, Three-Dimensional ; Female ; Heart Failure ; therapy ; Humans ; Male ; Middle Aged ; Treatment Outcome

OBJECTIVETo observe the psychological stress status in patients with acute coronary syndrome (ACS) and stable angina pectoris (SA).

METHODSThe intensity of social psychological stress and the serum levels of IL-6, CRP and ICAM-1 were determined in patients with ACS (n = 67) and SA (n = 33).

RESULTS(1) The percentage of patients with psychological stress was significantly higher in ACS than that in SA group (78.8% vs. 21.2%, P < 0.01). (2) The serum levels of CRP [(14.82 +/- 5.07) g/L vs. (8.78 +/- 4.34) g/L], IL-6 [(101.7 +/- 22.2) ng/L vs. (71.1 +/- 23.5) ng/L] and sICAM-1 [(1.41 +/- 0.47) mg/L vs. (0.82 +/- 0.37) mg/L] were significantly higher in psychological stress group than those in non-psychological stress group (all P < 0.05). Serum CRP [(18.91 +/- 3.12) g/L vs. (6.20 +/- 2.46) g/L], IL-6 [(114.6 +/- 15.2) ng/L vs. (56.4 +/- 15.8) ng/L] and sICAM-1 [(1.67 +/- 0.39) mg/L vs. (0.63 +/- 0.28) mg/L] levels in ACS group were significantly higher than those in SA group (all P < 0.01).

CONCLUSIONHigher psychological stress was associated with higher risk of ACS and increased serum inflammatory cytokines.

Acute Coronary Syndrome ; blood ; psychology ; Adult ; C-Reactive Protein ; metabolism ; Humans ; Intercellular Adhesion Molecule-1 ; blood ; Interleukin-6 ; blood ; Male ; Middle Aged ; Stress, Psychological

This paper reports the determination of the drug antibody ratio in an antibody-drug conjugate with two methods, i.e. LC-MS and UV/VIS, and to provide a reliable method to scientifically evaluate and effectively control the drug antibody ratio. Deglycosylated sample was analyzed with C4 column followed by MS, and the number of conjugated drugs in the antibody was determined by the molecular weight increase due to the addition of different number of drugs to the antibody, and then drug antibody ratio was calculated by weighted average of different number of drugs conjugated to the antibody. Optical density at 252 and 280 nm was measured with UV/VIS, and due to the difference of extinction coefficients between the antibody and the drug, the drug antibody ratio was calculated from linear equation with two unknowns. The drug antibody ratio was 3.21 and 3.25 respectively measured by the two methods, and the results were similar with the two methods. Our study indicated that both methods, LC-MS and UV/VIS, could be applied to the analysis of drug antibody ratio of the antibody drug conjugate.
Antibodies ; analysis ; chemistry ; Gas Chromatography-Mass Spectrometry ; methods ; Glycosylation ; Immunoconjugates ; analysis ; chemistry ; Maleimides ; analysis ; chemistry ; Molecular Weight ; Pharmaceutical Preparations ; analysis ; chemistry ; Spectrophotometry, Ultraviolet ; methods

Objective This paper aimed to report the rate of suicide attempts among adolescent and young students of eight cities in China, and to examine the impact of related psychosocial factors on suicide attempts. Methods The investigation was obtained in senior and junior high school and college students in eight cities of China. All of the 17 622 participants were recruited to complete anonymous questionnaires regarding their experiences of suicide attempts and related psychosocial factors. Results Overall, 2.2 percent of the respondents reported that they had experienced attempted suicide during the previous year. Scores on high anxiety (OR=2.61,95%CI: 1.64-4.16),high depression (OR=2.05, 95%CI: 1.35-3.10),suicide idea (OR= 14.21, 95%CI: 9.88-20.43),suicide plan(OR=3.50,95%CI:2.59-4.73) were revealed as common risk factors while the whole time span being accompanied by mothers during juvenile years (OR=0.38, 95% CI: 0.20-0.74) seemed to be protective factor for suicide attempts through multivariate logistic regression analysis. Conclusion Our results showed that emotional symptom, idea and plans for suicide were statistically associated with suicide attempted in high school and college students.