OBJECTIVE:To discuss the choice and the route for the realization of brand strategy feasible to Chinese pharmaceutical industry(CPI)in the current and future competition environment.METHODS:Reasons for the lack of influential brands were analyzed based on the current practical situation of CPI,then some countermeasures were put forward.RESULTS&CONCLUSIONS:Knowing the importance of brands and strengthening brands construction are the key issues for CPI to consider in the market competition,however,establishing national or even international brands is the long-term target of the future development of CPI.

Through the improvement on determination of rhTNF-? bioassay, we established the formal procedure . Methods; We choose L929 cell for rhTNF-? bioassay determination. To identify the influence of rhTNF-? on the growth of HL-60 cells, various concentration and effecting time were studied. Results: The results showed that the inhibition of rhTNF-? on HL-60 cells based on dose and time. Northern Dot Blot and DNA electrophoresis showed c-myc gene was surpressed, and DNA was damaged by rhTNF-?. Conclusion: Autometic calculation is responsible for bioassay of rhTNF-?. rhTNF-? could inhibit the proliferation of HL-60 cells and its mechanism is different from doxirubicin.

Objective: To establish National Standard of rhTNF-? complying with the requirements of research and quality control. Methods: National Standard of rhTNF-? were assayed against international standard (NIBSC) of rhTNF-? by cytotoxicity bioassay (L929 ) in vitro. The collaborative study has been carried out among four laboratories. Results: Based on the statistical analysis the results show that mean pf 95 % confidence interval is 3 289 ~ 4 266 IU per ampoule; the 95 % reference range is 1 735 ~ 8 087 IU per ampoule. Based on this study, the potency of rhTNF-? standard is defined as 3500 IU per ampoule. Stability tests indicate that the bioactivity has not been changed significandy under the storage of four different temperautres for 27 monthes. Conclusion: The preparation of rhTNF-? met the requirement of quality and could be used as a National Standard.

Objective To study the antibacterial effect of borneol, honey, gentamicin in Ruchuang Bingmi hydropathic compress agent.Method Staphylococcus aureus(S.aureus), Escherichia coli(E.coli), Pseudomonas aeruginosa(P.aeruginosa) were used to produce the third phase of infection decubitus animal models, respectively.Vaseline as the control group, divided into 12 groups:Vaseline-S.aureus, Vaseline-E.coli, Vaseline-P.aeruginosa;Honey-S.aureus,Honey-E.coli,Honey-P.aeruginosa;Borneol -S.aureus;Borneol -E.coli;Borneol-P.aeruginosa;Gentamicin-S.aureus;Gentamicin-E.coli;Gentamicin-P.aeruginosa;6 rabbits in each group.Honey, borneol, gentamicin was made into a gauze in treatment for decubitus .Organizations do strain identification and colony counts was observed before and after taking the treatment.ResuIts Borneol group ( F =11.059,P<0.01).,there was differences of each groups count cultured by borneol;Time ×Strains(F=11.281,P=0.009),there was no significant interaction between the two groups;gentamicin(F=7.99,P=0.000),gentamicin culture showed a difference in each group count;Time ×Strains(F=12.531, P<0.07),interaction between the two groups showed significant.Borneol has no antibacterial effects on P.aeruginosa , had a certain antibacterial activityon S.aureus and E coli;gentamicin had good antibacterial effect on the three kinds of bacteria, and against P.aeruginosa was particularly significant.ConcIusion The antibacterial activity of gentamicin is better than single herbs borneol in Ruchuang Bingmi hydropathic compress agent, honey has no antibacterial effect.

To establish a detection method of oncolytic adenovirus/p53 and standard of quality control, human telomerase reverse transcriptase (hTERT) promoter, CMV fusion promoter containing hypoxia reaction element (HRE) and p53 gene were identified by vector DNA restriction enzyme digestion and PCR analysis. The result conformed that all modified regions were in consistent with theoretical ones. Particle number was 2.0 x 10(11) mL(-1) determined by UV (A260). Infectious titer was 5.0 x 10(10) IU mL(-1) analyzed by TCID50. In vitro p53 gene expression in human lung cancer cell H1299 was determined by ELISA, and A450 ratio of nucleoprotein in virus infection group to control group was 5.2. Antitumor potency was evaluated by cytotoxicity assay using human lung cancer cell A549, and the MOI(IC50) of this gene therapy preparation was 1.0. The tumor cells targeted replication ability of recombinant virus was determined by TCID50 titer ratio of filial generation virus between human lung cancer cell A549 and human diploid epidermal fibrolast BJ cells after infected by virus with same MOI. TCID50 titer ratio of tumor cell infection group to normal cell infection control group was 398. The IE-HPLC purity of virus was 99.5%. There was less than 1 copy of wild type adenovirus within 1 x 10(7) VP recombinant virus. Other quality control items were complied with corresponding requirements in the guidance for human somatic cell therapy and gene therapy and Chinese pharmacopeia volume III. The detection method of oncolytic adenovirus/p53 was successfully established for quality control standard. The study also provided reference for quality control of other oncolytic viral vector products.

Objective To study the role of TLR2 and TLR4 signal transduction in RAW264.7 monocyte-macrophages stimulated by Aspergillus fumigatus conidia,and to investigate the expression of TLR2 signal transduction after silencing gene of TLR4.Methods Macrophages were randomly divided into normal group ( N group),normal+stimulated with Aspergillus fumigatus conidia ( N +Af group ),normal + transfected with TLR4-siRNA [ TLR4 (RNAi) group ],normal+transfected with TLR4-siRNA +stimulated with Aspergillus fumigatus conidia[ TLR4(RNAi) +Af group].RT-PCR and Western blot were used to assay expression levels of TLR2,TLR4,MyD88 mRNA and pro-inflammatory cytokines TNF-α protein when macrophages were stimulated 12 h by Aspergillus fumigatus conidia after tranfected 24 h with TLR4-siRNA by technology of RNAi.Results ( 1 ) Compared with N group,the expression of TLR2,TLR4,MyD88 mRNA and TNF-αprotein in N+Af group significantly increased before silencing gene of TLR4.(2) Silencing efficiency of macrophates was up to 83％ after transfected with TLR4-siRNA.(3)The expression of TLR2,MyD88 mRNA in TLR4 (RNAi) group significantly decreased contrast with normal group.Meanwhile the expression of TLR2,MyD88 mRNA and TNF-α protein also obviously reduced in TLR4(RNAi) +Af group when compared with N +Af group.Compared with TLR4 (RNAi) group,the expression of MyD88 mRNA in TLR4 (RNAi) +Af group significantly increased.However,the expression of TLR2 mRNA and TNF-α protein have no significant change after silencing gene of TLR4.Conclusion Signaling pathway of TLR2 and TLR4 in macrophages was activated by given stimulus of Aspergillusfumigatus conidia and exerted the effect of anti-Aspergillus fumigatus spores stimulation through the release of pro-inflammatory cytokines TNF-α.Meanwhile,silencing gene of TLR4 down-regulate the effect of TLR2 signal transduction in RAW264.7 cells to anti-Aspergillus fumigatus conidia stimulation,and it found that TLR4 played an more important role by contrast with TLR2.

Structure of a recombinant chimeric anti-CD20 IgG1 monoclonal antibody was verified by the application of high-performance liquid chromatography-mass spectrometry (HPLC-MS)and N-terminal sequencer. Molecular masses, N-terminal sequences and peptide maps of the antibody treated with different reagents and enzymes were measured. Results indicate that the amino acid sequences of light and heavy chains and 10 disulfide bonds were consistent with theoretical structure. By comparison of molecular masses and peptide maps for the fully glycosylated and deglycosylated samples, the N-linked glycosylation site was identified. The method is simple, rapid, precise, and could be referred to the quality control and structure determination of other IgG1 products.

Purpose To investigate the clinicopathological and immunohistochemical features of primary renal lymphomas ( PRL) , and to discuss the diagnosis, differential diagnosis, treatment and prognosis of the tumors. Methods Clinical data of 19 patients with PRL from January 2005 to October 2014 were retrospectively analyzed. Result The 19 patients in this study, there were 11 males and 8 females and the age ranged from 37 to 85 years old (averaged 55). Patients were mainly presented with unilateral renal masses, with lumbodynia as the main symptom. 13 patients underwent nephrectomy, 6 patients underwent renal biopsy and 17 patients received CHOP or R-CHOP chemotherapy. All of them were diagnosed as non-Hodgkin’ s lymphoma, with 14 cases of diffuse large B cell lym-phoma (DLBCL) (73. 684%, 14/19), 4 cases of B cell small cell lymphoma (21. 053%, 4/19), and 1 cases of T cell lymphoma (5. 263%, 1/19). Follow-up information was available in 15 patients. 12 were still alive and survived for 1~78 months, while the other 3 were dead with 1 case who died of cerebral infarction, and survived for 3~38 months ( averaged 23 months) . Conclusion PRL is an extranodal lymphoma which is rare in kidney and is often misdiagnosed as renal carcinomas due to its nonspecific clinical manifestations. The diagnosis of PRL can be confirmed by histopathological examination, immunohistochemistry and molecular analy-sis. The majority of the lymphomas are B cell lymphomas and most of them are DLBCL. The recommended treatment is surgery com-bined with chemotherapy and the prognosis is associated with the age, clinicopathological characteristics, tumor types and treatment.

AIMTo establish quality control requirements and methods for recombinant adeno-associated virus(rAAV) type 2/human blood coagulation factor IX (rAAV-2/hFIX).

METHODSIdentification of rAAV genome fragments, potential contaminants including wild type AAV(wtAAV) and helper virus, were detected by PCR. Purity of rAAV-2/hFIX was analyzed by cation-exchange HPLC and SDS-PAGE. Virus partical numbers were performed by dot blot assay. hFIX expression was demonstrated by ELISA and potency of hFIX was verified by APTT.

RESULTSIdentity of rAAV-2/hFIX was proved. Residues of wtAAV and helper virus were conformed to requirements. Purity of rAAV-2/hFIX were more than 98%. Partical numbers of rAAV-2/hFIX were more than 1.0 x 10(15) VG.L-1. hFIX expression was more than 20.0 micrograms.L-1. hFIX potency was verified by APTT following rAAV-2/hFIX injected to FIX gene knockout mice, potency results conformed to requirements.

CONCLUSIONThe methods and requirements had been established for quality control of rAAV-2/hFIX.

Animals ; Dependovirus ; genetics ; Factor IX ; biosynthesis ; genetics ; Female ; Gene Transfer Techniques ; Genetic Vectors ; genetics ; Genome, Viral ; Humans ; Male ; Mice ; Mice, Knockout ; Quality Control ; Random Allocation ; Recombinant Proteins ; biosynthesis ; genetics

The design and development of multi-dimensional ultrasonic reconstruction and visualization system (MURVS) have been described in the present paper. This system is basically composed of four modules: the data input/output module, image segmentation and arrangement module, multi-dimensional reconstruction module, and the dynamic visualization module. At first, some algorithms used in the system are introduced by the authors, including the AVI segmentation algorithm, three-dimensional interpolation algorithm, and volume rendering algorithms. Then the key questions of techniques to be discussed are: how to design the main modules, how to solve the dynamic visualization problem, and how to implement the system. The experiments indicate that MURVS is able to reconstruct all three-dimensional data fields in one cardiac cycle of a patient within 4 seconds, and dynamically display the motion of the heart. It allows the medical professionals to select different parameters when observing the reconstructed results. This is very helpful for medical professionals to reach more accurate diagnoses of their patients' diseases.
Adult ; Algorithms ; Echocardiography, Three-Dimensional ; methods ; Female ; Humans ; Image Processing, Computer-Assisted ; methods ; Mitral Valve Stenosis ; diagnostic imaging