Objective To produce prokaryotic recombinant protein glyceraldehyde-3-phosphate dehydrogenase of Clonorchis sinensis (CsGAPDH), analyze its enzyme activity and immunological function. Methods The recombinant CsGAPDH was purified according to the protocol of GST?Bind~TM kit and was digested with thrombin proteinase and eluted with wash buffer. The BALB/c mice were inoculated with the purified protein. The antisera collected from the mice were used to detect the titres of IgG antibodies by ELISA, and Western blotting was used to identify the specificity of the antisera with the purified CsGAPDH. S-P immunohistochemistry method was used to confirm the expression and distribution of CsGAPDH in adult Clonorchis sinensis with the polyclonal antibodies from immunized BALB/c mice. The CsGAPDH catalytic activity was evaluated employing the conventional substrate glyceraldehydes-3-phosphate (3-GAP). Results SDS-PAGE showed a single purified protein band. Gel scanning analysis revealed that the protein purity of CsGAPDH was 90%. ELISA analysis showed an increased IgG value. S-P immunohistochemistry analysis demonstrated that the recombinant plasmid pGEX-4T-1-GAPDH expressed and distributed in muscle cell membrane of immune mice. Western blotting result suggested that CsGAPDH protein contained essential epitopes with high antigenic activities. This protein CsGAPDH could catalyzed 3-GAP with enzymatic active unit of 2 872 U min~-1ml~-1. Conclusion The recombinant protein CsGAPDH shows a proper enzymatic activity and immunogenicity.

Objective:To detect the contents of hyaluronic acid(HA),Laminin(LN) and probe their role in the pathologic course of chronic renal failure(CRF).Methods:The serum HA and LN from 37 patients with CRF and 40 healthy adults were measured by RIA.T test was used in data(x?s) analysis.Results:The HA and LN in CRF serum evidently increased than those of normal controls,respectively(P

Objective To construct prokaryotic recombinant plasmids of transcriptional co-activator (TC) gene of Clonorchis sinensis, express and purify the recombinant protein and analyze its biological function. Methods A pair of primers was designed according to the known sequence of TC gene. The TC gene fragment was amplified by PCR. After purification and digestion with BamHⅠ and SalⅠ , the TC gene was connected to the prokaryotic expression vectors, pGEX-4T-1 and pET30a(+). By cloning target gene into these vectors, pGEX-4T-1 and pET30a(+), prokaryotic recombinant plasmids of TC gene were constructed and transferred into E.coli BL21. The positive expressed recombinants were detected by SDS-PAGE and Western blotting. Immobilized metal (Ni 2+ ) chelation affinity chromatography was used to purify His-TC produced by the expression of the recombinant protein pET30a(+)-TC. Results The recombinant plasmids, pGEX-4T-1-TC and pET30a(+)-TC, were constructed successfully. SDS-PAGE testified that the molecular weight of the recombinant protein was correct. Western blot analysis of GST-TC recombinant protein testified that the recombinant protein could be recognized by immunized rabbit serum, which means the protein is GST-immune active and the clone can express recombinant Clonorchis sinensis antigen. After affinity chromatography of the pET-TC protein, there was only one protein band with expected size on the SDS-PAGE gel. Conclusion The TC gene was screened from cDNA library of adult Clonorchis sinensis, cloned, expressed and purified. The purified protein of TC gene will be of importance for further research on the biological function of the gene.

A slight halophilic denitrifying bacteria(designated GYL)was screened from the activated sludge which was used to treat high-salinity wastewater.According to the results of morphological observation,physiological and biochemical test,sequence analysis of the 16S rDNA,strain GYL was identified as Halo-monas sp..This strain could survive at 10% salinity and the optimal salinity range for growth was 2%~7%.The suitable pH value for growth was 7.5~8.5 and sucrose was the most effective carbon source.The nitro-gen removal efficiency exceeded 80% when the temperature ranged from 25?C to 30?C.Meanwhile hetero-trophic nitrification characteristics of this strain were measured.Results showed that this strain was able to realize SND and ammonia removal rate was 98.3%.It showed that this strain could perform the whole proc-ess of bacteria denitrification independently.

Ancient literature of ethnic medicine , as a concentrated expression of the ethnic culture of Chinese medicine , has a distinct regional and national character . In this paper , the questionnaire , interview , literature and other methods , were applied to elaborate the research situation of ethnic medicine ancient literature in Sichuan province from the aspect of the literature resources investigation, excavation arrangement, and etc. Ac-cording to the different national medical status differences , we pointed that the national pharmaceutical develop-ment ignored literature research . And we proposed recommendations on how to protect and use the ethnic medicine ancient literature . This paper provided a scientific basis for the further promotion of ethnic medicine literature excavations work in Sic huan province .

Objective To evaluate diagnostic value of X-ray computed tomography(CT) neck masses in children.Methods Clinic,pathologic diagnosis and CT scans of 26 patients from Jan.2004 to Jun.2006 with neck masses which location,density,edge and near construction were reviewed retrospectively.Results Seven cases in neck anterior area,13 cases in neck lateral area,6 cases in other areas or unconfirmed.Eight cases with lower density than cervical muscles,2 cases with same density,2 cases with higher density,14 cases with mixed density.Inflammatory masses were observed in 13 cases,congenital malformation in 11 cases,thyroiditis in 1 case,pharyngeal tumor-like proliferation in 1 case.Conclusion CT scan is the best choice of diagnosis of neck masses in children,and is valuable to diagnosis associated with history and physical examination.

High altitude polycythemia (HAPC) had become one of the main common chronic diseases, which had seriously threatened the health of Plateau people. In the Tibetan medicine classic bookSi-Bu Yi-Dian, there were recordings on HAPC treatment methods and medications, which had the unique advantages of identified therapeutic effect with little side effect. This article analyzed Tibetan medicine in the prevention and treatment of HAPC from aspects such as etiology and pathogenesis, clinical treatment advantages and modern innovation study. Questions were also raised on lacking of standardization on HAPC clinical effectiveness, as well as Tibetan medicine compound material basis and action mechanisms were unclear. It was proposed that based on the inheritance of Tibetan medicine theoretical basis and clinical therapeutic effect, the Tibetan medicine original thinking should be combined with modern science and technology, in order to strengthen the analysis of ancient literature collection in HAPC treatment and data mining in medication experiences. The clinical treatment standards and medication plan should be standardized. Methods of systems biology, such as metabolomics, can be used in the further study of the scientific connotation of HAPC treatment by Tibetan medicine.

Objective To investigate the dynamic changes of the spinal cord during neck flexion in Hirayama disease for diagnosis.Methods MRI examinations in neutral neck position and a fully flexed neck position were performed on 18 cases of Hirayama disease and 31 young normal control subjects.We measured an antero-posterior diameter(APD)and transverse diameter(TD)of the cervical cord at the superior margin of the C6 vertebral body for each position,and investigate the dynamic changes.The different in frequency of these findings between the control and patient groups was examined by means of the x~2 test.The group means were compared by independent-sample t-test.Significance was defined as P

20% improvement in 4/7 primary indices.Sharp declines in several parameters were noticed during 3 to 6 months.Pain in 9 patients disappeared.There was also a statistically significant decrease in TNF ? transcripts in the PBMC.Conclusion Thalidomide is a reasonably promising drug for treating resistant ankylosing spondylitis and its biological mechanism is related to the gene expression suppression of proinflammatory cytokine TNF ?.

Objective To investigate the effect of recombinant adenovirus mediatied human endostatin (rAD-GFP-ES)on rats with collagen typeⅡinduced arthritis(CIA),and explore the mechanism of inflamma- tion and cytokines inhibition on rats CIA.Methods The rAD-GFP-ES was amplified and purified.The model of rat CIA was induced by intradermal injection of typeⅡcollagen combined with complete Freund's adjuvant(CFA). On the second day after the injection,the therapeutic administration of rAD-GFP-ES(1?10~(11)pfu?kg~(-1)?week~(-1)?4 weeks)were performed to the rats.The mean arthritis index(AI)was scored every week since then.The relative concentrations of ES,IL-I?,TNF-?in sera collected at the fourth week were evaluated by western blotting. Results①The titer of the purified rAD-GFP-ES and rAD-GFP was 6.6?10~(12)pfu/ml and 4.8?10~(12)pfu/ml,re- spectively(A_(260nm)/A_(280nm)＞1.3).②The concentration of ES in sera of the group treated with rAD-GFP-ES was 2.4-lold higher compared to the normal group.③The mean arthritis index of the group treated with rAD-GFP- ES was much lower than that of the model group.The administration of rAD-GFP-ES could significantly de- creas the production of IL-1?and TNF-?in sera.Conclusions①The rAD-GFP-ES is efficiently expressed in vivo.②The rAD-GFP-ES has an inhibitory effect on the arthritis index of rat CIA.③IL-1?and TNF-?are involved in the pathogenesis of RA.The rAD-GFP-ES has an inhibitory effect on the expression of IL-1?and TNF-?in rat CIA.