To establish a coupling model for pharmaceutical industrial cluster and circular economy, and study the factors of coupled associations between them. The factors of coupled associations between pharmaceutical industrial cluster and circular economy were analyzed,and an effective route for sustainable development of our domestic pharmaceutical industry was seeked. The pharmaceutical industrial cluster and circular economy were coupled through scales, information, costs and intergrowth,which is beneficial to achieve scale effect, spread facilitate technical, save environmental costs and enhance the cohesion among the clusters and to achieve intensive development for pharmaceutical industry.

Breast cancer stem cells (BCSC)are a class of breast cancer cells that have the capacity of selfrenewal and can differentiate into different cell lineages. These cells, with the ability of high tumorigenesis, invasion and metastasis, play a important role in breast cancer metastasis, recurrence and treatment resistance. The treatments targeting on breast cancer stem cells are very important for improving the efficacy of clinical therapy.

Objective: To fuse Mycobacterium tuberculosis protective antigen gene with mice ubiquitin gene, constructing antigen ubiquitin system. Methods: Mice ubiquitin cDNA was amplified by RT PCR from mice testicle,and 4 antigen genes were obtained by PCR from cultured Mycobacterium tuberculosis . Ubiquitin and 4 antigen genes were linked by flexible adaptor respectively and the fusing genes were cloned into pVAX vector.The recombinant plasmids were digested with endonuclease and sequenced.Then the recombinant plasmids were transfected into COS7 cells and the expression was assayed by ELISA. Results:Ubiquitin and 4 antigen genes were 0.2,0.3,0.7,1.0,1.65 kb in length by agarose electrophoresis. Endonuclease digestion of the recombinant plasmids indicated that the fusion genes were correctly inserted into pVAX vector. Sequencing results of fusion nucleic acid vaccines were identical to those in GenBank.The recombinant plasmids expressed in COS7 cells. Conclusion: Four Chinese Mycobacterium tuberculosis protective antigens ubiquitin systems are successfully constructed and can be expressed in eukaryotic cells. This may provide a basis for diagnosis and therapy of tuberculosis.

Objective To investigate the resistance of Salmonella enterica serotype Typhimurium (STM) to quinolone. Methods A total of 33 salmonella typhimurium resistant to Ciprofloxacin in stool of outpatients from July 1st. 2004 to Oct 31st was isolated. in four hospitals in Wuhan. Antimicrobial susceptibility test was conducted by the Agar Dilution method. All of the strains were studied for the occurrence of mutations in the genes coding for QRDR by PCR and REP-PCR. Results The sequencing of QRDR of these genes in highly quinolone-resistant mutants (MICs of 4 to 16?g/ml) of the 25 STM revealed the presence of gyrA mutation, and inserted base in the gyrB and parE genes were found only in 5 STM. With aid of REP-PCR, the 32 strains were divided into 9 profiles which were correlated to the antibiotic susceptibility spectrums. Conclusions The resistance of STM from community acquired infection in WuHan is very severe. The resistance mechanism was associated with mutations of the quinolone resistance-determining regions. It suggests an outbreak in the community.

Objective: To screen out DNA binding proteins specifically recognizing CpG immunostimulatory sequence (ISS) for further investigating the molecular mechanisms of ISS. Methods: Yeast one hybrid system was adapted in screening a human bone marrow cDNA library using 4 copies of ISS as bait. The ISS binding activity of the positive clone was confirmed by electrophoretic mobility shift assay (EMSA). Results: Four dual positive colonies were obtained, two of them encoded proteins with unknown functions. The other 2 encoded light chains of immunoglobulin with amino sequences homology to anti DNA Ab and HBsAb respectively. EMSA showed HBsAb specifically bound to CpG ODN at pH6.4 and pH 5.8. Conclusion: HBsAb may have ISS specific DNA binding activity.

Objective To clone and express the gene encoding ESAT antigen of Mycobacterium tuberculosis (chinese stain). Methods Genome of mycobacterium tuberculosis was extracted. Then, the full length cDNA encoding ESAT protein was amplified by PCR and cloned into prokaryotic expressing vector pGEX5T and sequenced. pGEX5T ESAT was expressed in k802 Ecoli, and the expressed protein was determined by western blot using the sera from ten patients with tuberculosis. Results One specific band of 0.3kb or so was obtained and sequencing result was identical to that reported from Genbank. The expressed protein could be specifically recognized by the sera from tuberculosis patients. Conclusions The full length cDNA of Mycobacterium tuberculosis (Chinese strain) ESAT protein was cloned and expressed successfully, which will be helpful in further studies on diagnosis and treatment of tuberculosis.

Objective: The fibrogenicity of fur dust was studied in rat lung tissues. Methods: Intratracheal instillation of fur dust, morphologic examination of lungs and analysis of collagen content were performed in Wistar rats. Results: Morphologic examination revealed that the earliest changes consisted of alveolar edema, increased numbers of intraalveolar macrophages, and marked thickening of interalveolar septa with mixed cellular infiltrate. After sixth months, there was moderate thickening of the alveolar walls and the peribronchioli. After 12 months, interstitial positive fibrosis of the alveolar wall and the peribronchioli were weakly seen. In the carding dust group (silica content 17.6%), interstitial nodules were observed composed of fibroblasts, reticular fibers, and collagen fibers. Electron microscopic examination also showed that alveolar walls became thickened and collagen fiber bundles were seen around bronchioles and small vessels in the carding groups after 12 months. At all stages of analysis, the collagen content in lungs of the fur dust groups was significantly higher than that of the control group. Conclusions: Our study suggested that fur dust might induce weak interstitial fibrosis in the lung.
Dust ; Collagen ; interstitial ; month ; seconds

Objective: To explore the effects of ulinastatin on the level of MIF in rats with acute necrotic pancreatitis. Method: 52 healthy Wister rats were randomly divided into three groups: normal control group(group C, 12), ANP group(group A, 20)and UTI group(group U, 20). Severe acute pancreatitis rat model in group A were induced by injection of 315 % sodium taurocholate through retrogradely common biliopancreatic ducts via papilla duodeni. After inducing the rat model of ANP through the way above, rats in group U were treated by ulinastatin through portal vein injection. Pancreas and duodenum were only flipped after opening abdominal cavity in group C. Then rats were killed at 3rd, 6th ,12th ,24th hour after operation respectively. Cut the belly open at once, and draw blood in postcava. The levels of serum MIF were determined with ELISA. Blood amylase was detected through biochemistry instrument. Resected pancreas tissues was scored according to the standard of Kusske. Result: Compared to the normal control group, the level of serum MIF , blood amylase and histopathological scores were significantly increased in ANP group, P

Objective To study the changes of proteome expression in brain tissues from rats with severe traumatic brain injury(sTBI). Methods Total protein of brain tissues were obtained at days 3,7 and 14 for two-dimensional gel electrophoresis to screen and identify differential protein spots.Results We screened 17 differential protein spots that were involved in cellular metabolism,stress and inflammatory reaction. Conclusion Some differential proteins involved in sTBI can be found by twodimensional gel electrophoresis.

Objective To locally inject human umbilical cord blood (HUCB) mesenchymal stem cells (MSCs) to rat traumatic brain injury (TBI) model to investigate expression of neural markers and neurological functional improvement. Methods HUCB-MSCs were labeled by bis-benzimide for over 24 hours and stereotactically transplanted into the brain of the rats. All rats were divided into four groups, ie, sham injury group, TBI group, control (TBI + PBS) group and treatment (TBI + MSCs) group, Im-munohistochemical methods and immanofluorescence staining were used to observe the survival, migration and differentiation of the transplanted cells. The neurological functional improvement was evaluated by u-sing the neurological severity score (NSS). Results There existed a large number of MSCs survived in local region of the brain that received transplants, when some MSCs differentiated into neurons or astro-cytes and expressed the neurocyte markers including NSE and GFAP around the grafted site. Treatment group had significantly improved scores compared with sham injury group, TBI group and control group. Conclusions HUCB-MSCs transplantation can potentially improve neurological functional after TBI and may be a good alternative to bone marrow cells for stem cell transplantation or cell therapy.