BACKGROUND:Increasing evidence suggests that Sanguis draxonis is of great significance for treating pressure sores, burns and ulcers.
OBJECTIVE:To observe the influences of Sanguis draxonis on vascular endothelial growth factor, epidermal growth factor, substance P and hydroxyproline in rats undergoing tissue-engineered skin transplantation and to verify its promotion of wound repair.
METHODS:The tissue-engineered skin transplantation model of rats were established. Rats in treatment groups were given external application, single oral use of 0.1 g/(kg·d) Sanguis draxonis and combined use, respectively. No treatment was given in control group. After continuous treatment for 7 days, the expression levels of vascular endothelial growth factor, epidermal growth factor, substance P and hydroxyproline in skin tissue were determined.
RESULTS AND CONCLUSION:Compared with the control group, the levels of vascular endothelial growth factor, epidermal growth factor, and hydroxyproline were significantly increased (P<0.05 or P<0.01), while substance P had no change in the treatment groups (P>0.05). These results demonstrate that Sanguis draxonis can promote tissue-engineered skin to repair skin wound by upregulating the expression levels of vascular endothelial growth factor, epidermal growth factor, and hydroxyproline.

Objective To study the immunogenicity of Streptococcus pneumoniae pneumolysin DNA vaccine in Rhesus macaques. Methods The deletion of the gene sequence encoding for the 11 amino acids at the carboxyl terminal of pneumolysin (PN) from its wild type gene (pn) by PCR resulted in a mu-tant pneumolysin gene (pnd). The wild type pn gene encoding PN and the mutant gene (pnd) encoding PND were cloned into pVAX1 vector respectively and then tested as Ppn and Ppnd DNA vaccines. The PN and PND proteins were purified from recombinant E. coli. Rhesus macaques were immunized by intramuscu-larly (i.m.) injection of Ppn or Ppnd DNA vaccine with electroporation (EP). Results Because of the deletion of the gene sequence encoding for the eleven amino acids at the carboxyl terminal of the PN from pn gene, the recombinant PND antigen lost its hemolytic activity while its antigenicity was remained. The spe-cific humoral immunity against pneumolysin was induced by injecting monkey with 500 μg DNA followed by EP. Boosting the Ppn or Ppnd DNA/EP primed animals with corresponding recombinant protein, PN or PND, evoked strong immune response at about 4 fold increase in the antibody titer. Conclusion Specific antibody responses were induced in the monkeys by DNA vaccination and electroporation. The immunogenic-ity of the DNA vaccines were significantly enhanced when PN or PND protein boost was applied 10 d after third DNA vaccination.

Objective To establish a stable and useful method for culturing human osteoclast-like cells in vitro,and investigate the effect of 1?,25-(OH)2D3,M-CSF and PGE2 on osteoclasts differentiation,proliferation and activation so as to lay the foundation for further study of the biological mechanism for tooth movement.Methods The HCMNC were isolated and cultured in 24-well plate with coverslips and human dentine slices.The experiment group was cultured with 1?,25-(OH)2D3,M-CSF and PGE2,respectively,while the control group was not.The liquid was changed every 3 days and the whole culture process lasted for 7 days.The phase contrast microscopy and TRAP staining were adopted to identify osteoclast-like cells.Results On the 3rd day the monocytes began to fuse and on the 7th day positive multinucleated cells could be seen with TRAP staining,but absorption pit was not formed on the dentin slices.The group with 1?,25-(OH)2D3 had the largest number of osteoclast-like cells.Conclusion After the monocytes in UCB are cultured by 1?,25-(OH)2D3,M-CSF,PGE2 induction,they can turn into TRAP(+) multinucleate osteoclast-like cells,the 1?,25-(OH)2D3 10-8mol/L being the most effective.

Objective: To observe the effect of polysaccharide ingredients from six TCM complex prescriptions on releasing the cytokine(CK) level of mouse celiac macrophages (M?), to explore the possible mechanism and direct the extractionand separation of active components for above compounds. Methods: The total polysaccharide ingredients of six complex prescriptions were prepared: Siwu Decoction, Sijunzi Decoction, Liuweidihuang Decoction, Guizhi Decoction, Longdanxiegan Decoction and Yupingfeng Pulveres, then the mouse celiac M? were incubated together in 96 holes board. Furthermore, levels for the latter to release CK, including IL-1?, IL-6, IL-8 and TNF-?, were measured. Results: Polysaccharide ingredients of each complex prescription could obviously accelerate mouse celiac M? to release one or more CK, and the e ects had concentration otherness (P

AIM:To establish macrophage mannose receiver Model(MMR),and use it to screen active component with mannose receiver(MR)as target from traditional Chinese herbs.METHODS:The mouse abdominal macrophages was hatched with D-mannose and D-galactose of the different concentration,and the flow cytometry and fluorescence microscope were used to measure the antagonistic effect of M-FITC-BSA(Mannose-fluorescein isothiocyanate-bovine serum albumin)with D-mannose and D-galactose.After the MMR screening model was established to screen MR union ingredients of polysaccharide ingredients from six compound prescriptions,such as Siwu Decoction and so on.RESULTS:Both of measuring methods showed that when D-mannose concentration increased the relevance ratio of M? marked with M-FITC-BSA decreases(P

Object A novel microwave-heated extraction (MHE) method was studied for the extraction of glycyrrhizic acid from Glycyrrhiza uralensis Fisch. Methods Several factors, such as temperature, time and microwave power were investigated and the appropriate MHE conditions were obtained from the orthogonal test. Under the optimum conditions, the optimal solvent was selected and the MHE was compared with ultrasonic extraction, leaching at room temperature and Soxhlet extraction,. Results The optimum conditions of MHE is extracting for another 40 min in 0.5% ammonia water after heated to 60 ℃ by microwave of 2 000 W. Yield of glycyrrhizic acid was about equal to that of Soxhlet extraction for 4 h, and that of leaching at room temperature for 44.3 h. Conclusion The MHE method is fast, efficient, energy-saving and high-selective, which is recommendable to the application to active compounds extraction from Chinese herbal medicines.

Objective To analyze the clinical features of juvenile dermatomyositis (JDM) combined with soft-tissue calcification. Methods Forty-seven patients with JDM combined with soft-tissue calcification (soft-tissue calcification group) were retrospectively analyzed, and they were contrasted with 89 patients with non-calcification (non-calcification group). Results The rates of Gotton signe, muscle contracture and joint dysfunction in soft-tissue calcification group were significantly higher than those in non-calcification group:87.23% (41/47) vs. 43.82% (39/89) and 68.09% (32/47) vs. 21.35% (19/89), and there were statistical differences (P<0.05). The dosage of glucocorticoid (conversion of prednisone measuring more than 1.5 mg/kg), rate of using immunodepressant, level of creatine kinase in soft-tissue calcification group were significantly lower than those in non-calcification group:17.02%(8/47) vs. 68.54%(61/89), 25.53%(12/47) vs. 88.76%(79/89), (566.45±240.41) U/L vs. (1 680.12±656.50) U/L, and there were statistical differences (P<0.05). Conclusions The patients with JDM combined with Gotton signe are more prone to soft-tissue calcification. The rate of muscle contracture and joint dysfunction in soft-tissue calcification patients is significantly higher than that in non-calcification patients. For the patients whose creatine kinase are not obviously elevated, they are more prone to soft-tissue calcification. Early active application of glucocorticoid and immunodepressant therapy can reduce or prevent the occurrence or development of late calcium deposition.

Objective To prepare the chitosan-potD nanoparticles and to evaluate its protective efficacy against pneumococcal nasopharyngeal colonization. Methods potD gene was amplificated from pneumococcal genome and was inserted into pVAX1 expression vectors to construct pVAX1-potD recombinant plasmid which was then transfected into 293T cell using LipofectAMINE 2000 to analyze transient potD gene expression in vitro by RT-PCR and Western blot. Chitosan-potD nanoparticles were freshly prepared by coacervation methods at each time and the characterizations of the nanoparticles were then evaluated. BALB/c mice were immunized with chitosan-potD, naked potD DNA or pVAX1 for 4 times at two-week intervals. Anti-PotD IgG, IgG1 and IgG2a levels in serum and IgA levels in nasal washes, bronchoalveolar lavage fluids (BALF) and middle ear lavages(MEL) were detected by indirect enzyme-linked immunosorbent assay (ELISA). IL-17A, IL-4 and IFN-γ levels in splenocytes were determined by double sandwich ELISA. Mice were intrannsally challenged with Streptococcus pneumoniae ATCC6303, and Pneumococci were recovered from the nasopharyngeal niche at the fifth day after challenge. Results potD gene was successfully amplificated by PCR and the sequence was confimed to be consistent with that in the Genbank. The pVAX1-potD recombinant plasmid was successfully constructed and was expressed in eukaryocytes in vitro. The mean size and zeta potential of chitosan-potD nanoparticles was 430 nm and + 20.5 mv, respectively. Chitosan-potD nanoparticles were not digested by DNase Ⅰ , while naked potD DNA was completely digested. The levels of antibodies inculding IgG, IgG1, IgG2a, IgA and cytokines including IL-17A, IL-4 and IFN-γ were significantly higher in mice immunized with chitosan-potD nanoparticles than mice with naked potD or pVAX1 ( P <0.05) only. More importantly, much less Pneumococci were recovered from mice immunized with chitosan-potD nanoparticles than the other groups(P <0.05). Conclusion Chitosan-potD nanoparticles significantly enhanced the immunogenicity and protection efficacy of DNA vaccines by intranasal immunization and could be used as a potential mucosal vaccine to prevent pneumococcal infection.

OBJECTIVE To prepare pneumolysin(Pn)by genetic engineering and thereby establish the basis for the study of vaccines against otitis media. METHODS A pair of primers including two restriction sites was designed based on the pneumolysin gene sequence reported by Walker in 1987. The pneumolysin gene was PCR-amplified from pneumococcal DNA. The resulting fragment, digested by restriction enzymes, was ligated into the vector PET-28a and then transformed into host cell E.coli JM109(DE3). RESULTS The sequence of the inserted pneumolysin gene was confirmed by DNA sequencing. CONCLUSION The pneumolysin gene was successfully cloned into the host cell.

Objective By culturing the osteoclasts together with the osteoblasts directly to investigate the effect of osteoblasts on the formation of mature osteoclasts.Methods The bone marrow mononuclear cells of rats were treated with 30?g/L M-CSF and 50?g/L RANKL and cultured for 6 days.Subsequently,the primary osteoblasts which were of the same quantity as the osteoclasts were co-cultured directly.In the co-culture system,we added the liquid containing 1,25-(OH)2D3 1?10-8mol/L and PGE2 1?10-6mol/L.The morphological observation,TRAP staining and pit staining were adopted to identify osteoclasts.Results When the osteoclasts were co-cultured with primary osteoblasts,the growth of osteoblasts had more preponderances.After staining,we could see more osteoblasts than osteoclasts.Conclusion The relationship between osteoblasts and osteoclasts is related to the relative quantities of the two cells.When osteoblasts outnumber osteoclasts,osteoblasts would inhibit the formation and differentiation of osteoclasts.