There is an insufficient supply in essential medicines, so the nation intends to reconsider the designated medicine production. But this will raise certain risks such as negative distribution, monopolies and weakening price signal. Thus the supporting methods should be proposed as below: improving the incentive of enterprises, the efficiency of designated products, and the government administration level to promote the development of the pharmaceutical market and improve the level of public health.

Objective To investigate the effect of matrix metalloproteinase(MMP-2) in the attack of exprimental autoimmune myositis(EAM) rats, and the effects of methylprednisolone on expressions of gene mRNA and protein of MMP-2. Methods The expression levels of MMP-2 in the peripheral blood, spleen lymphocytes and muscles were detected by using RT-PCR and immunohistochemistry techniques and was compared among EAM group,EAM with methylprednisolone treatment group (EAMM) and control group. Results ⑴ The degree of episode in EAMM group was lower than that of EAM group , and the infiltration of the inflammatory cells and the necrosis of the muscles were also mild in EAMM group. ⑵There was significant upregulation of the expression of the MMP-2 gene mRNA in the lymphocytes of peripheral bood and spleen of EAM rats as compared with that of the control,Elevation of the expression of MMP-2 protein in rats muscles tissues of EAMM group was observed obviously. However, the expression of the mRNA and the protein of MMP-2 was suppressed significantly in EAM group compared with control group. Conclusions The upregulation of the expression of MMP-2 may be correlated with the onset of EAM. Methylprednisolone may relieve the degree of the pathology of EAM by the downregulation of MMP-2 .

AIM　Diclofenac potassium sustained-release tablets were developed as a gel matrix for using twice daily and evaluated in vitro release characteristics. METHODS　The formulations were screened according to the designed in vitro release rate by the use of HPMC as the gel matrix and the hydrophobic retarding agent to modify the release. RESULTS　The release was properly characterized by the diffusion mechanism, influenced by pH of the media, slightly affected by the basket rotating speed, without the influences from the pressure exerted during the tabletting procedure. CONCLUSION　The selected formulation of diclofenac potassium sustained-release tablets could ensure the desired in vitro release rate. In addition, it proved a good manufacturing reproducibility.

Objective To study the relationship between sICAM 1,sVCAM 1,TNF ?,TNFR Ⅰ,TNFR Ⅱ and pathogenesis of vasculitis.Method The serum levels of TNF ?,TNFR Ⅰ,TNFR Ⅱ,sICAM 1 and sVCAM 1 were detected by ELISA and their levels between patients with vasculitis and controls were compared.Results Compared with the controls,the serum level of TNF ? was lower,the levels of TNFR Ⅰ,sICAM 1 and sVCAM 1 were higher in the patients with vasculitis,and there was no difference of TNFR Ⅱ between the two groups.Conclusion The low level of TNF ? suggests that this factor may not play a role in these patients,or its role was counteracted by its receptors—sTNFR,whose level was higher in patients with vasculitis.sICAM 1 and sVCAM 1 may play a significant role in the pathogenesis of vasculitis.

OBJECTIVETo study the molecular mechanism of extracts from Euodia rutaecarpa on hepatotoxicity in mice.

METHODTotally 30 KM mice were divided into 3 groups and orally administrated extracts from E. rutaecarpa for consecutively 15 days. The expressions of Erkl/2, CDK8, CK1e, Stat3 and Src were detected by Western blotting method.

RESULTThe extracts from E. rutaecarpa could up-regulated Erkl/2, CDK8 and CK1e expressions (P <0.01) and down-regulate Stat3 and Src (P <0.01).

CONCLUSIONThe molecular mechanism of E. rutaecarpa on hepatotoxicity may be correlated with Erkl/2, CDK8, CKle, Stat3 and Src signal molecules.

Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Down-Regulation ; Drugs, Chinese Herbal ; toxicity ; Evodia ; chemistry ; Female ; Fruit ; chemistry ; Gene Expression Regulation ; drug effects ; Liver ; drug effects ; metabolism ; Male ; Mice ; Plants, Medicinal ; Signal Transduction ; drug effects ; Specific Pathogen-Free Organisms ; Triglycerides ; blood ; Up-Regulation

er analysis of the survival showed that there was significant difference between patients with and without OH. Conclusion At the introductory phase of haemodialysis, OH is an independent predictor of survival rate in haemodialysis patients.

AIM:To express recombinant hCD154-GST fusion protein, to prepare anti-hCD154 monoclonal antibody, and to investigate the effect of anti-hCD154 monoclonal antibody on graft rejection. METHODS AND RESULTS: Total RNA was prepared from human peripheral blood mononuclear cell (PBMC) activated with 10ng/mL PMA and 1 ?g/mL PHA for 8h, the total RNA was reversetranscribed to cDNA. The entire coding region and a part of the 3'non-coding regions were amplified by PCR using a pair of primers designed and synthesized according to the sequence of human CD154 gene from gene bank. The amplified product, a 820bp DNA fragment was cloned into pGEX-4T-1 plasmid expressing glutathione S-transferase(GST). The cloned insert was identified by double digestion of the cloned pGEX-4T-1 plasmid with retriction enzymes BamHⅠand EcoRⅠ.The fusion protein expression plasmid of PGEX-4T-1/hCD154 was constructed, then transformed to E coli BL21. The human CD154-GST fusion protein expression was induced by IPTG in BL21. The expression of recombinant 26kD GST and 55kD human CD154-GST fusion protein were confirmed by SDS-PAGE. CONCLUSION: We have express the recombinant human CD154-GST fusion protein. The expressed hCD154-GST fusion protein will be used to prepare anti-hCD154 monoclonal antibody, to investigate the role of anti-CD154 monoclonal antibody on graft rejection.

AIM:To obtain the light chain region(VL) and heavy chain region(VH) genes from the hybridoma cell line and analyse their sequence for construction of the engineering antibody against hCD154. METHODS: In this research ,total RNA was extracted from the hybridoma cell line 7E8, which secretes McAb against hCD154, and subjected to reverse transcription. The VL gene and VH gene were amplified by PCR, cloned into puc18 vector and sequenced by Sanger's dideoxymediated chain-termination method. RESULTS: The VL cDNA of 7E8 McAb consists of 341 bp encoding 113 amino acid residues. Compared with mouse Ig database, the VL region is in accord with the characterization of DNA sequence present in the mouse Ig Vk region , it belong to mouse V?2 light chain. The VH cDNA of 7E8 McAb consists of 354 bp encoding 118 amino acid residues. Compared with mouse Ig database, the VH region is in accord with the characterization of DNA sequence present in the mouse Ig VH region. CONCLUSION: The DNA squence analysis showed that the cloned genes code the light and heavy chain variable region of mouse respectively.

Linoleic acid (C18∶2n-6) and ?-linolenic acid (C18∶3n-3) are found widely in fungi, plants and some lower animals. However, they can not be synthesized in mammals due to lack of △12 and ?-3 fatty acid desaturases. To enable endogenous production of essential fatty acids in mammalian cells, here the stable expression of a Caenorhabditis elegans gene FAT-2 encoding △12 fatty acid desaturase in CHO cells was reported. First, the FAT-2 coding sequence was cloned by RT-PCR. To facilitate high level synthesis of heterogeneous protein, the codon usage of the fatty acid desaturase genes was optimized according to the codon preference of mouse by site-directed mutagenesis, 2 synonymous mutations were introduced into FAT-2 gene by overlapping PCR. The codon-modified gene was finally fused to pBudCE4.1 vector (Invitrogen) under the control of CMV promoter. The expression vector pBudCE-FAT2 was linearized with NheⅠ, and then transfected CHO cells, the cells were under Zeocin selection for nine days and then propagated, then the transfected cells were harvested. The genome and total RNA were isolated for PCR and Norhern blot ananlysis. The results revealed that FAT-2 gene has been integrated into the genome of CHO cells and expressed properly. Fatty acids of total cellular lipids were analyzed by gas chromatography. The results indicate that the expression and function of △-12 fatty acid desaturase resulted in accumulation of linoleic acid. The levels of linoleic acid in transgenic cells were 2.4-fold higher than those in wild-type cells. The moderate linoleic acid in CHO cells was derived from cell culture media uptaken by cell membrane. The results demonstrate that a heterogenous desaturase gene can function well in mammalian cells and prove that transgenic approach is an efficient strategy for changing fatty acid composition of mammals.

OBJECTIVETo investigate clinical outcomes of Kirschner wire as blocking screws combined with interlocking intramedullary nail internal fixation in treating tibial metaphyseal fractures (AO 43A).

METHODSFrom March 2011 to June 2012, 9 patients with tibial metaphyseal fractures were treated with blocking screws Kirschner wire combined with interlocking intramedullary nail, including 7 males and 2 females aged from 23 to 54 years old with an average of 37.4. Postoperative complications, X-ray were observed, AOFAS scoring were used to evaluate function after operation at 12 weeks.

RESULTSAll patients were followed up from 6 to 40 weeks (mean 20.1), and healed at stage I. No serious swelling, infection and skin necrosis occurred. No fracture instability and displacement appeaered at 4 and 8 week after operation. AOFAS score was (95.2±4.6) at 12 weeks after operation and 7 patients gained excellent result and 2 patients good.

CONCLUSIONKirschner wire as blocking screws with interlocking intramedullary nail for treatment of tibial metaphyseal fractures can fix well and perform simply.

Adult ; Bone Screws ; Bone Wires ; Female ; Fracture Fixation, Intramedullary ; methods ; Humans ; Male ; Middle Aged ; Tibial Fractures ; surgery