The high performance liquid chromatography-fluorescence micelle assay (HPLC-FMA) method for the content determination of polysorbate 80 in monoclonal antibody drugs was validated to study its applicability and transferability between various laboratories, and the feasibility to be included in the Chinese Pharmacopoeia. Both J.T. Baker and Nanjing Well-sourced polysorbate 80 was used in the collaborative validation of polysorbate 80 content analysis in seven different laboratories. The results show that when the protein concentration was no more than 20 mg·mL^-1 and the concentration of polysorbate 80 ranged from 0.05 to 0.5 mg·mL^-1, the method had good specificity. The recovery rates of the spiked samples ranged from 92.20% to 117.70% for J.T.Baker and from 93.90% to 117.20% for Nanjing Well. The intra-laboratory precision (%RSD) was less than 4.30% for J.T. Baker, and less than 2.60% for Nanjing Well, while the overall precision was less than 5.45% for J.T. Baker, and less than 6.70% for Nanjing Well. The linear correlation coefficient was more than 0.98 for J.T. Baker and more than 0.99 for Nanjing Well. The results of the collaborative validation prove that the HPLC-FMA method has good accuracy, precision, linearity, and specificity, and could be used for release control analysis of polysorbate 80 content in monoclonal antibodies across different laboratories.

Objective To discuss the frequent etiology of spontaneous sub-cortical hemorrhage and its diag- nosis.Methods The clinical materials of 79 cases of spontaneous sub-cortical hemorrhage were analyzed.Results 56% of the hemorrhage was caused by arterial-venous malformation.48% of the hemorrhage was caused by occult AVM.Conclusion AVM is the most frequent etiology of spontaneous sub-cortical hemorrhage.CTA plays a special role in its diagnosis.

AIMTo study and establish the fingerprint of Polygala tenuifolia by RP-HPLC.

METHODSThe fingerprint of P. tenuifolia was built by using Kromasil C18 as column and acetonitrile-0.05% phosphoric acid aqueous in gradient as mobile phase. The flow rate was 1.0 mL x min(-1), and the detecting wavelength was set at 318 nm. Total 14 batches of P. tenuifolia from different habitats were detected, and different commodities and preparative methods were compared.

RESULTSSamples from different habitats were of high similarities, and the qualities of "yuanzhitong" and "yuanzhirou" were better than that of "yuanzhigun", while the fingerprints of P. tenuifolia with different preparative methods have not obvious differences. Moreover, the standard fingerprint of P. tenuifolia was originated from the "Computer Aided Similarity Evaluation" software, and 29 common peaks existed in the fingerprint. Each peak in the fingerprint was separated very well under the above chromatographic condition, with good accuracy, stability and repeatability, satisfied to the requests of fingerprint.

CONCLUSIONThe RP-HPLC fingerprint method can be used in the quality control of P. tenuifolia.

Chromatography, High Pressure Liquid ; methods ; Ecosystem ; Hot Temperature ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Polygala ; chemistry ; Quality Control ; Reproducibility of Results ; Technology, Pharmaceutical ; methods

The objective was to identify some biochemical and physical properties for fusion protein IL6D24-PE40KDEL. Edman degradation, SDS-PAGE, peptide mass fingerprinting, Western blot and MTT were used for identification of the protein. The results showed that the sequence of N-terminus is Met-Ile-Asp-Lys-Gln-Ile, Met was added because of prokaryotic expression system; Western blot revealed that the purified protein could react with IL6 and PEA antibody. The purified protein IL6D24-PE40KDEL could kill the multiple myeloma cell lines U266 expressing high affinity IL6R, but it could not kill the cell lines CEM which not expressed IL6R; The molecular weight was 58.7 kD measuring by SDS-PAGE; peptide mass fingerprinting (PMF) confirmed that the construction of IL6D24-PE40KDEL was correct. A novel protein by Peptident database in EXPASY web site was identified. In conclusion, IL6D24-PE40KDEL is a new targeting protein with bioactivity of specific killing effect.
ADP Ribose Transferases ; chemistry ; metabolism ; pharmacology ; Amino Acid Sequence ; Blotting, Western ; Cell Line, Tumor ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Exotoxins ; chemistry ; metabolism ; pharmacology ; Humans ; Interleukin-6 ; chemistry ; metabolism ; pharmacology ; Molecular Sequence Data ; Pseudomonas aeruginosa ; genetics ; metabolism ; Recombinant Fusion Proteins ; chemistry ; metabolism ; pharmacology ; Sequence Analysis, Protein ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVETo observe the role of interleukin (IL) 21 alone, IL12 alone, and IL21 plus IL12 for inducing the antitumor activity of peripheral blood mononuclear cells (PBMCs) in patients with endometrial cancer.

METHODSPBMCs were isolated from peripheral blood in patients with endometrial cancer in vitro, and kept the culture with low-level IL2. IL2-stimulated PBMCs were cocultured under different conditions (with anti-IL21 antibody, IL21 alone, IL12 alone, or IL21 plus IL12) for 72 h. The cytotoxicity of PBMCs was then examined by lactate dehydrogenase(LDH) released assay. CD4(+) CD25(+) FOXP3(+) regulatory (Treg) cell and CD4(+) IL17A(+) T-helper (Th17) cell proportion were determined with flow cytometry. Cell proliferation and apoptosis were measured by cell counting kit-8(CCK-8)assay and flow cytometry, respectively.

RESULTSIn comparison to control group, both IL21 and IL12 significantly enhanced the cytotoxicity of PBMCs. The IL21 plus IL12 group had superior effect to IL21 alone and IL12 alone. IL21 and IL12 significantly decreased the percentages of Treg cells and the rate of PBMCs apoptosis. IL21 or IL12 had no significant effect on the differentiation of Th17 cells and the proliferation of PBMCs.

CONCLUSIONSIL21 and IL12 can enhance the cytotoxicity of PBMCs in patients with endometrial cancer, which can be further strengthened with treatment of IL21 plus IL12. Such effects may be achieved by inhibiting the differentiation of Treg cells and the apoptosis of PBMCs, but not by the differentiation of Th17 cell.

Adult ; Aged ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endometrial Neoplasms ; immunology ; pathology ; Female ; Humans ; Interleukin-12 ; pharmacology ; Interleukins ; pharmacology ; Leukocytes, Mononuclear ; drug effects ; immunology ; pathology ; Middle Aged ; T-Lymphocytes, Regulatory ; immunology

The purpose of this study was to detect the minimal residual disease (MRD) in peripheral blood of newly diagnosed patients with acute myeloid leukemia (AML) on day 8 of induction chemotherapy and analyze the correlation between day 8 MRD (D8RD) and therapeutic effectiveness. 29 adult patients (13 males and 16 females, aged 16 - 75 years, median 41 years) with AML diagnosed and treated in West China Hospital from September 2009 to June 2010 were analyzed and followed up in the study. The leukemia-associated aberrant immunophenotype (LAIP) of all the patients were detected by multiparameter flow cytometry (FCM) before therapy. The level of MRD in the peripheral blood at day 8 of induction chemotherapy was detected by FCM based on the LAIP. The overall survival curve was drawn by calculation using Kaplan-Meier method using, and the comparison between different groups was carried out by Log-rank test. The results indicated that after first course therapy, the levels of peripheral D8RD in 7 out of 29 AML cases were lower than 0.01% (negative group), and that in another 22 cases were higher than 0.01% (0.08% - 55%, positive group). The sex, age, WBC, LDH, percentage of bone marrow blasts at diagnosis in these groups were not statistically different. 6 cases achieved CR (86%) in D8RD negative group, and also 6 cases achieved CR (27%) in D8RD positive group, CR rate in D8RD negative group was higher than in D8RD positive group (86% vs 27%, P < 0.05). The median follow-up of 29 cases lasted for 15 months; the 1-year overall survival rate of D8RD negative and D8RD positive groups was 100% and 39.4%, respectively (P < 0.01). It is concluded that MRD level in peripheral blood at day 8 of induction chemotherapy is an early index to predict clinical efficacy of induction therapy in AML.
Adolescent ; Adult ; Aged ; Early Diagnosis ; Female ; Flow Cytometry ; Humans ; Leukemia, Myeloid, Acute ; blood ; drug therapy ; mortality ; Male ; Middle Aged ; Neoplasm, Residual ; diagnosis ; drug therapy ; mortality ; Prognosis ; Survival Rate ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the influence of chronic prostatitis/chronic pelvic pain syndromes (CP/CPPS) on male health.

METHODSWe conducted investigations among 200 cases of CP/CPPS, aged 20 -59 years and randomly selected from 650 out-patients, using an anonymous questionnaire including such items as the patients'age, height, body weight, disease duration and treatment, the results of EPS and semen determinations, and the scores on NIH-CPSI, IIEF-5 and Symptom Checklist (SCL-90).

RESULTSAccording to the 198 valid questionnaires collected, CPSI scores showed 56 (28.3%) cases to be mild, 98 (49.5%) moderate and 44 (22.2%) severe. Sperm density and motility (grade a + b) were reduced in 38 (19.2%) and 47 (23.7%) cases, respectively. IIEF-5 revealed ED in 41 (20.7%) cases, of which 58.5% were mild. Psychological abnormality was found in 94 (47.5%) cases, including anxiety, depression, sensitiveness and so on. NIH-CPSI scores were positively correlated with psychological symptoms and ED (r = 0.25, P < 0.05; r = 0.12, P < 0.05), but not significantly related with sperm density and motility (P > 0.05).

CONCLUSIONThe influence of CP/CPPS is obvious on male psychology but mild on penile erection and not significant on semen. Integrated physiological and psychological therapy is recommended for the treatment of CP/CPPS.

Adult ; Chronic Disease ; Humans ; Male ; Middle Aged ; Pelvic Pain ; psychology ; Penile Erection ; Prostatitis ; psychology ; Quality of Life ; Surveys and Questionnaires ; Young Adult

Chemical constituents of ethyl acetate extract of Artemisia rupestris were isolated and purified by various chromatographic methods, including silica gel, ODS and Sephadex LH-20. Their structures were elucidated on the basis of spectroscopic data analysis. 12 compounds were separated from A. rupestris and their structures were identified as 5,4'-dihydroxy -3,6,7-trimethoxy flavone (1), R-(-) -vestitol (2), tricin (3), chrysoeriol (4), 3-indole carboxylic acid (5), esculetin (6), apigenin (7), luteolin (8), trans-caffeic acid (9), casticin (10), chrysosptertin B (11) and artemetin (12). Compound 2 was obtained from the genus Artemisia for the first time, and compounds 1-6 and 9 were separated from this plant for the first time.
Artemisia ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Mass Spectrometry ; Molecular Structure

OBJECTIVETo study the effects of MTA1 small interfering RNA (siRNA) on the anchorage-independent growth and anoikis of prostate cancer cell line PC-3.

METHODSAfter transfection of human prostate cancer PC-3 cells by MTA1 siRNA, we detected the expression of the MTA1 gene by real-time PCR and Western blot, the anchorage-independent growth of the cells by clone formation in soft agar, and their anoikis by DNA fragmentation assay and flow cytometry.

RESULTSCompared with the control group, MTA1 siRNA transfection significantly decreased the mRNA and protein levels of MTA1, inhibited the anchorage-independent growth of the PC-3 cells, and induced their anoikis, all in a dose- and time-dependent manner (r = 0.935, P = 0.001; r = 0.901, P = 0.0005; r = 0.916, P = 0.0003).

CONCLUSIONMTA1 siRNA can inhibit the anchorage-independent growth of prostate cancer cells by inducing their anoikis.

Anoikis ; genetics ; Gene Expression Regulation, Neoplastic ; Histone Deacetylases ; genetics ; Humans ; Male ; Prostatic Neoplasms ; genetics ; RNA, Small Interfering ; Repressor Proteins ; genetics ; Transfection ; Tumor Cells, Cultured

Background::The dermoscopic features of rosacea have already been reported. However, the current findings are incomplete, and little is known about phymatous rosacea. Hence, this study aimed to summarize and compare the dermoscopic features and patterns of three rosacea subtypes (erythematotelangiectatic [ETR], papulopustular [PPR], and phymatous [PHR]) in the Chinese Han population and to evaluate whether these features differ with patients’ genders, ages, and durations.Methods::Dermoscopic images of 87 rosacea patients were collected in non-polarized and polarized dermoscopy contact modes at 20-fold magnification. Dermoscopic features, including vessels, scales, follicular findings, and other structures, were summarized and evaluated.Results::The reticular linear vessels and red diffuse structureless areas of ETR were distinctive. For PPR, red diffuse structureless areas, reticular linear vessels, yellow scales, follicular plugs, and follicular pustules were typical dermoscopic criteria. The common dermoscopic features of PHR were: orange diffuse structureless areas, linear vessels with branches, perifollicular white color, orange focal structureless areas, and white lines. The following features statistically differed among the three rosacea subtypes: reticular linear vessels ( P < 0.001), unspecific linear vessels ( P= 0.005), linear vessels with branches ( P < 0.001), yellow scales ( P = 0.001), follicular plugs ( P < 0.001), perifollicular white color ( P < 0.001), red diffuse structureless areas ( P = 0.022), orange diffuse structureless areas ( P < 0.001), red focal structureless areas ( P = 0.002), orange focal structureless areas ( P = 0.003), white lines ( P < 0.001), follicular pustules ( P < 0.001), and black vellus hairs ( P < 0.001). Conclusions::The dermoscopic patterns of ETR are red diffuse structureless areas and reticular linear vessels. For PPR, the pattern comprehends combinations of red diffuse structureless areas, reticular linear vessels, yellow scales, follicular plugs, and follicular pustules. Meanwhile, PHR is characterized by remarkable orange diffuse structureless areas, linear vessels with branches, perifollicular white color, orange focal structureless areas, and white lines.