OBJECTIVETo evaluate the clinical efficacy and safety of Qiangji Recipe (QR) in ad- junctive treatment of axial undifferentiated spondyloarthritis (axuSpA) through a four-week open study.

METHODSFifty-four axuSpA patients of Shen-deficiency Du-channel cold syndrome (SDDCS) in line with inclusive criteria were recruited and assigned to the treatment group and the control group according to random digit table, 27 in each group. Patients in the control group took Celecoxib Capsule (0.2 g each time, twice per day). Patients in the treatment group additionally took QR (consisting of Herba Epimedii 15 g, antler glue 15 g, Cibotium Barometz 15 g, eucommia bark 20 g, dipsacus asper 10 g, two toothed achyranthes root 15 g, drynaria 15 g, Taxillus Chinensis 20 g, ground beetle 10 g, scorpion 5 g, wild celery 10 g, notopterygium incisium 10 g, cow-fat seed 10 g, white mustard seed 6 g, and licorice root 6 g, one dose per day, twice daily). The therapeutic course for all was 4 weeks. The Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), the Bath Ankylosing Spondylitis Functional Index (BASFI), the Bath AS Metrology Index (BASMI), total body pain and spinal pain, patient and physician global assessment on a four-point scale, the Ankylosing Spondylitis Quality of Life (ASQoL), erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) were measured before and after 4 weeks of treatment. The primary end point in this study was the proportion of patients with a 20%improvement response accord- ing to the ASAS International Working Group Criteria (ASAS 20 responders) at week 4.

RESULTSTotally 50 patients completed this trial, 26 in the treatment group and 24 in the control group. Improvement of BASDAI, BASFI, BASMI, ASQoL, ESR, and CRP was shown in both groups after treatment. Better effect was shown in the treatment group in all indices except ESR and BASMI after treatment (P < 0.05, P < 0.01). Twenty cases (accounting for 76.92%) in the treatment group achieved ASAS 20 response at week 4, while 12 cases (accounting for 50.00%) in the control group achieved ASAS 20 response at week 4 (P < 0.05). No obvious adverse reaction occurred in the two groups.

CONCLUSIONQR combined Celecoxib Capsule showed better effect in treating axuSpA patients than using Celecoxib Capsule alone.

Blood Sedimentation ; C-Reactive Protein ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Pain ; Quality of Life ; Spondylitis, Ankylosing ; drug therapy

Objective To prepare natural anti-keratin IgM monoclonal antibody. Methods Spleen cells of BALB/c mice raised in specific pathogen free conditions were directly fused with Sp2/0 myeloma cells. The hybridoma supernatants were tested by ELISA using pre-extracted keratin. The natural IgM obtained was further identified by immunochemistry and immunoblot methods. Results The cell fusion rate was about 60% without pre-immunization. About 14% supernatants reacted with the keratin antigen. Three hybridoma strains secreting natural IgM monoclonal antibody against keratin were obtained. The immunochemistry results showed that the natural anti-keratin IgM was able to bind to epidermis, sebaceous gland, hair follicule, and muscle tissues. Conclusion B lymphocytes in normal BALB/c mice spleen can produce natural antibody against kerain.

Objective To conduct extensive quality control tests on Madin-Darby Canine Kidney ( MDCK) cells used for the production of influenza vaccine .Methods Tests for characteristics , extraneous agents, endogenous agents and tumorigenicity were performed on MDCK cells according to Chinese Pharma -copeia Book III .Cell lysate and DNA of MDCK cells were tested for oncogenicity in the light of new interna -tional requirements .Results The MDCK cells extracted from canis were adherent cells with an epithelial morphology, whose average number of chromosome was 80±1.No bacteria, fungi and mycoplasma contami-nation were detected . The detection for extraneous and endogenous virus showed that there was no nonspecific virus causing cytopathic effect , hemadsorption , hemagglutination or animal death .Tests for re-verse transcriptase , bovine viruses and canine viruses were all negative .Each nude mouse was injected with 107 viable cells to observe their tumorigenicity .Twelve weeks after cell injection , no node was found at the injection site and in large organs by gross anatomy .There was no significant difference between test group and negative control group .The test for tumorigenicity of viable cells was negative .Cell lysate and cellular DNA collected from equivalent amount of cells were respectively injected into nude mice , and no node forma-tion was found.There was no significant difference between the test cells and negative controls in pathology indicating that the tested MDCK cells were non-oncogenic .Conclusion It showed the possibility of using MDCK cells for the production of influenza vaccine .

Objective To investigate the children's body environmental Se and T-2 toxin level in their staple food in Kaschin-Beck disease(KBD)relative active regions in Aba state of Sichuan province in 2008.Methods We took X-ray photograph of the right hand on children aged 7-13 years in 48 villages from 11 counties in Aba state.The relative active regions of KBD were chosen according to the X-ray result and historical status of KBD.The children's urine and hair,drinking water and their staple food werr sampled.Selenium contents in urine,hair,water and food samples were determined by naphthalene fluorescence,and T-2 toxin in staple food samples were detected by ELISA kits.Results In 2145 X-ray films,66 films were positive,and the children's KBD positive rate was 3.08%(66/2145).The KBD positive rate was respectively 10.98%(29/264)and 8.52%(19/223)in Maerkang county,Jinchuan county and it was 0.75%(3/400)in Rangtang county,historically serious endemic area.The selenium content in urine of children aged 7-13 years in Maerkang county,Jinchuan county and Rangtang county was (10.41±4.67), (10.11±3.65), (8.42±2.68)μg/g Cr, respectively, there was no statistical difference among three counties(F=0.901, P>0.05). The selenium content in hair of children aged 7-13 years in Maerkang county[(0.18±0.04)mg/kg] was lower than that in Jinchuan county[(0.21±0.04)mg/kg, P<0.05].The selenium content in water in Jinchuan county [(0.225±0.124 )μg/L ] was lower than that in Maerkang county and Rangtang county[(0.320±0.092), (0.339±0.105)μg/L, all P<0.05]. The selenium content in staple food in Jinchuan county(0.0033 mg/kg) was lower than that in Maerkang county and Rangtang county(0.0258,0.0137mg/kg, Z=-6.146,-3.042, all P<0.017). The T-2 toxin level in flour in three counties was 19.60,17.95,26.25 ng/g,respectively,there was no statistical difference among three counties(X2=5.623, P>0.05).The T-2 toxin level in grain Maerkang county (10.72 ng/g) was higher than that in Jinchuan county and Rangtang county (3.74,3.30 ng/g, Z=-6.315,-4.407,all P<0.017). T-2 toxin contamination in flour was more severe than that in grain (Z=-6.690,-5.493,-3.676, all P<0.05). Conclusions In 3 relative active KBD regions of Aba state,the children's selenium nutritional status and the T-2 toxin contamination level in their staple food is consistent with the distribution of KBD.

To silence the expression of K-RASAsn12 in human pancreatic cancer cell line by vector-based RNAi(RNA interference) technique,two single-strand DNA sequences encoding mutant-specific shRNA (short haipin RNA) for K-RASAsn12 were synthesized and then inserted into pSilenCircle. The recombinant plasmid was called pSC-K-RASAsn12. According to the same method, pSC-GFP encoding shRNA for GFP was gained. Both recombinant plasmids were transfected into human pacreatic cancer cell line AsPC-1 and BxPC-3. The expression level of K-RASAsn12 was detected by semi-quantitative RT-PCR and Western blot. The result indicated that the recombinant plasmid edcoding mutant-specific shRNA for K-RASAsn12 can inhibit significantly the expression of K-RASAsn12 without affection of wild-type K-RAS(K-RASWT)in Human Pancreatic Cancer Cell Line.

BACKGROUNDHeat shock protein 70 (HSP70) is expressed highly in epithelial tumours associated closely with human papillomavirus 16 (HPV16) infections. However, evidence about the direct relationship between HSP70 expression and HPVs infections are still lacking. In the present study, we examined the expression of HSP70 in keratinocytes introduced with HPV16 E6/E7 oncogenes.

METHODSStable transfected cells were established by transfection of the plasmids pLXSN16E6/E7 into cultured primary keratinocytes and subsequently selected by plasmid specific selection antibiotic (G418) at the required concentration. The expression of HSP70 in pLXSN16E6/E7 transfected keratinocytes was determined by Western blot. The correlation of HSP70 expression and E6/E7 transfection was further confirmed by doubly labelled immunofluorescent staining.

RESULTSCompared to non-transfected keratinocytes, there was a significant trend for higher levels of HSP70 in pLXSN16E6/E7 transfected keratinocytes. Doubly labelled immunofluorescent staining experiment showed that the co-localization of HPV16 E6/E7 and HSP70 in transfected keratinocytes was observed and increased expression of HSP70 was strongly associated with the transfection of HPV16 E6/E7.

CONCLUSIONSOur studies demonstrated increased levels of HSP70 proteins in keratinocytes stably transfected by HPV16 E6/E7 oncogenes. It suggests that the expression of HSP70 is modulated by HPV16 E6/E7 proteins, which may be involved in HPV16 E6/E7 induced immortalization.

Cells, Cultured ; HSP72 Heat-Shock Proteins ; biosynthesis ; Humans ; Keratinocytes ; metabolism ; Oncogene Proteins, Viral ; genetics ; Papillomavirus E7 Proteins ; Repressor Proteins ; genetics ; Transfection

Objective To investigate the clinical value of 18 F?FDG PET/CT in staging multiple myeloma ( MM) and evaluating the glucose metabolic activity of MM. Methods A total of 25 MM patients ( 13 males, 12 females, age:39-67 years) from May 2010 to April 2015 were enrolled in this retrospective study. The SUVmax of each patient was recorded. D?S plus staging according to 18 F?FDG PET/CT was com?pared with the traditional D?S staging. The SUVmax and the percentage of plasmacytes of bone marrow of phase Ⅲ and non?phase Ⅲ ( phaseⅠand Ⅱ) according to D?S plus staging were compared. Two?sample t test and Wilcoxon rank sum test were used to analyze the data. Results In 25 MM patients, the range of SUVmax of lesions was 1.8-12?0 and the mean value was 5.15±2.74. According to D?S staging, the numbers of patients with phase Ⅰ,Ⅱ andⅢwere 7, 4 and 14, respectively. While the numbers were 3, 1 and 21 by D?S plus staging. Based on the D?S plus staging system, stages of 7 patients ( 28%, 7/25 ) were changed. According to the D?S plus staging system, the SUVmax between phaseⅢand non?phaseⅢpatients was significantly different (5.75±2.54 vs 3.00±0?70; t=2.12, P<0.05), while the percentage of plasma?cytes of bone marrow between the 2 groups had no significant difference ( 17. 50%( 4. 25%-41. 75%) vs 11?15%(10.25%-36.57%);z=0.05, P>0.05). Conclusion 18F?FDG PET/CT is of clinical importance for MM staging and metabolic activity assessment of MM.

OBJECTIVETo explore the methods for labeling CDTPA-coupled CD45 monoclonal antibody (mAb) with yttrium-90 ((90)Y) for potential acute myeloid therapy.

METHODSCD45 mAb was labeled with (90)Y by CDTPA and the labeling rate, radiochemical purity, final specific activity, and immunological activity of the mAb were detected.

RESULTSWith the optimal molar ratio of CDTPA/Ab at 20:1, the labeling rate was 95%, radiochemical purity 99.8%, and final specific activity 1.9 mCi/mg. This conjugate was stable in vitro with comparable immunological activity in comparison with unlabeled CD45 mAb.

CONCLUSION(90)Y-CDTPA-CD45 mAb possesses good properties as an ideal targetting therapeutic agent for acute leukemia.

Anhydrides ; chemistry ; Antibodies, Monoclonal ; chemistry ; immunology ; Humans ; Immunoconjugates ; chemistry ; immunology ; Isotope Labeling ; methods ; Leukocyte Common Antigens ; immunology ; Pentetic Acid ; chemistry ; Radiopharmaceuticals ; chemical synthesis ; chemistry ; immunology ; Yttrium Radioisotopes ; chemistry

OBJECTIVETo investigate the effect of bortezomib alone and in combination with arsenic trioxide on apoptosis of HL-60 cells.

METHODSHL-60 cells were treated with bortezomib alone or in combination with arsenic trioxide for 12 to 48 h and the cell proliferation was analyzed with MTT assay, and cell apoptosis detected by DNA gel electrophoresis, fluorescence microscopy and flow cytometry.

RESULTSAt the concentrations of 10 to 50 nmol/L, bortezomib effectively inhibited HL-60 cell proliferation, and induced cell apoptosis. A 12-hour bortezomib treatment at 10 nmol/L was sufficient to induce cell apoptosis, and prolonged treatment or increased concentration significantly increased HL-60 cell apoptotic rate. Combined treatment of the cells with bortezomib (10 nmol/L) and arsenic trioxide (15 micromol/L) resulted in an even higher cell apoptosis rate than that induced by the respective agent alone.

CONCLUSIONBortezomib can induce HL-60 cell apoptosis in a time- and dose-dependent manner, and a synergistic effect can be observed of bortezomib and arsenic trioxide in apoptosis induction.

Animals ; Apoptosis ; drug effects ; Arsenicals ; administration & dosage ; pharmacology ; Boronic Acids ; administration & dosage ; pharmacology ; Bortezomib ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Drug Synergism ; Electrophoresis ; Flow Cytometry ; HL-60 Cells ; Humans ; Oxides ; administration & dosage ; pharmacology ; Pyrazines ; administration & dosage ; pharmacology ; Time Factors