OBJECTIVE: To establish the quality standard of Buqi tongmai capsule.METHODS:Salviae miltiorrhiza,Ligusticum chuanxiong,Borneolum Syntheticum,Citrus aurantium were identified qualitatively by TLC.The content of astragaloside A was analyzed quantitatively by TLC scanning method.RESULTS: The specificity of S.miltiorrhiza,L.chuanxiong,Borneolum Syntheticum and C.aurantium could be analyzed qualitatively by TLC.The linear range of astragaloside A were 0.274 2～4.934 7 ?g (r=0.998 9) with an average recovery of 96.56%(RSD=2.70%,n=5).CONCLUSION: Established quality standard can be used for the quality control of Buqi tongmai casuple.

Objective:To explore the changes of chemotherapy sensitivity of breast cancer MCF-7/ADM cells after Hiwi gene targeting silence, and to illuminate the role of Hiwi gene in resistant mechanism of breast cancer MCF-7/ADM cells.Methods:The MCF-7/ADM cells were transfected with Hiwi control,siRNA1 Hiwi gene and siRNA2 Hiwi gene as Hiwi control group, Hiwi siRNA1 group and Hiwi siRNA2 group.The expression levels of Hiwi gene mRNA and protein in breast cancer MCF-7/ADM cells were detected by Real-time PCR and Western blotting methods to judge the transfection rates in various groups.After transfection,the breast cancer MCF-7/ADM cells in various groups were treated with the different concentrations(0,0.1,0.5 and 1.0 mg·L-1) of adriamycin, and the cell resistant sensitivities were detected by MTT method.Results:Compared with control group(0 mg·L-1 adriamycin), the expression levels of Hiwi gene mRNA and protein inMCF-7/ADM cells in Hiwi siRNA1 and Hiwi siRNA2 groups were significantly decreased (P<0.01).Compared with control group, the survival rates of Cells in Hiwi siRNA1 and Hiwi siRNA2 groups were significantly decreased after treated with different concentrations of adriamycin,and the survival rates in Hiwi siRNA1 and Hiwi siRNA2 groups were significantly decreased(P<0.01);they were (48.15±6.28)% and (41.73±6.17)% when the concentration of adriamycin was 1 mg·L-1,and the sensitivities to the adriamycinwere obviously enhanced(P<0.01).Conclusion:The target silencing Hiwi genes in MCF-7/ADM cells is efficient.The sensitivity of MCF-7/ADM cells to adriamycin restores after Hiwi gene silencing.

OBJECTIVE: To optimize the preparation technology of Buqi tongmai capsule.METHODS: Orthogonal experiment of exeracting Radix Astragali was conducted with crude drugs soaking time,extracting times,amount of water added and decocting duration as factors and the content of astragaloside as index.Orthogonal experiment of exeracting Salvia miltiorrhiza,Ligusticum chuanxiong,Fructus Aurantii Immaturus,Carthamus tinctorius was conducted with crude drugs soaking time,amount of water added and decocting duration as factors and the content of tanshinol as index.RESULTS:Water was adopted for extraction and the concentration of alcohol for precipitate was 70 percent.The optimal technology of exeracting Radix Astragali was to soak the crude drugs for 30 min,decoct for 90 min in 10 times of water for 2 times.The optimal technology of exeracting S.miltiorrhiza,L.chuanxiong,Fructus Aurantii Immaturus,C.tinctorius was to decoct the crude drugs for 90 min,8 times of water for 2 times.CONCLUSION:The preparation processes is feasible,stable and meet the demands in the clinic.

Objective To introduce the method and to evaluate the therapeutic effect of collagenase chemonucleolysis for treatment of cervical disc herniation.Methods 92 patients with cervical herniated discs were selected from January 2002 to December 2004.The procedure was guided by DSA and the puncture was defined from C_(6~7) or C_7-T_1 extradural cavity.Collagenase(1200~2400 u) was injcted in the herniated extradural cavity through the micrcatheter.Results The procedure of 88 cases was successful.80 cases were followed up from 6 to 12 months.The effect showed that 70 cases(87.5%) were excellent or good.No serious complication occurred.Conclusion The method of collagenase chemonucleolysis for treating cervical disc herniation is safe and effective,it can be used in clinic.

Objective To discuss the clinical value of susceptibility weighted imaging (SWI) in the brain vascular malformations. Methods Imaging data of 34 patients with brain vascular malformations proved by digital subtraction angiography (DSA) or pathology obtained on Siemens Sonata 1.5T MR system were studied prospectively, and compared with those of conventional MRI (cMRI) and SWI. Results All 41 lesions of 34 patients with brain vascular malformations showed clearly by SWI. These patients were diagnosed by surgical findings or DSA. These nidus comprised 19 cavernous angiomas, 9 arteriovenous malformations and 6 cerebral venous malformations. Conclusion SWI should be used for clinical diagnosis of brain vascular malformations, and providing more complete and detailed information combining with other sequence.

To find anti-hypertensive lead drug, angiotensin converting enzyme (ACE) inhibitory peptides were synthesized and their effects on inhibiting ACE activity were investigated. ACE inhibitory peptides were synthesized via Fmoc solid-phase synthesis, isolated and purified through reversed phase high-performance liquid chromatography (RP-HPLC), and identified by mass spectrometry. A RP-HPLC analysis method was used to test ACE inhibitory activity in vitro of these ACE inhibitory peptides. Six octapeptides were successfully synthesized, and the analytical results of mass spectrum were consistent with their theoretically calculated data. Among these synthetic octapeptides, the anti-SARS (severe acute respiratory syndromes) octapeptide had the most obvious ACE inhibitory activity with an IC50 value of 3.4 x 10(-5) mol x L(-1). So octapeptide AVLQSGFR-OH (anti-SARS peptide) was found to be the strongest candidate for potential development as an anti-hypertensive drug and had the implication of further study.

BACKGROUND: Strontium-doped calcium polyphosphate (SCPP) is a new type of bone repair materials with good biocompatibility and controlled degradation. The preliminary studies of our group indicate their role in promoting angiogenesis,but its mechanism is unclear.OBJECTIVE: By co-culturing osteoblasts ROS17/2.8 with SCPP in vitro to observe cell proliferation and the secretion of vascular endothelial growth factor (VEGF).DESIGN, TIME AND SETTING: A contrast study was performed at the Laboratory of Tissue Engineering of Sichuan University from October 2008 to June 2009.MATERIALS: A series of calcium polyphosphate (CPP) respectively containing 0%, 1 %, 2%, 5%, 8%, and 10% Sr~(2+) were prepared. ROS17/2.8 osteoblastic cell strain was provided by Laboratory of Transplantation Immunity and Transplantation Engineering, West China Hospital, Sichuan University.METHODS: ①Preparation of cell scaffold complexes: The materials were placed in 24-well plates, then 300 μL cell suspension with a concentration of 2×10~7 cells/Lwas inoculated into each hole. These complexes were cultured for 14 days and the liquid was changed every two days. ②These complexes were measured by MTT assay to observe the proliferation of osteoblasts on the 1~(st), 3~(rd), 5~(th), 7~(th), 10~(th) and 14~(th) days, respectively. ③ The centrifugal supernatant of the complex cultured for seven days was measured by ELISA assay to check the secretion of VEGF.MAIN OUTCOME MEASURES: The proliferation of osteoblastic cells on SCPP and CPP was observed. The amount of VEGF protein secreting from osteoblastic cells was detected.RESULTS: The results of MTT showed that, compared with the CPP group, SCPP groups could promote the proliferation of osteoblasts, and 8% SCPP group was the best; ELISA results showed that, compared with the CPP group, SCPP groups could increase the amount of VEGF protein secretion, of which the promoting role of 8% SCPP was the most obvious (P < 0.05).CONCLUSION: When cultured with osteoblasts, SCPP can promote cell proliferation, and can significantly increase the secretion of VEGF; moreover, 8% SCPP is the best, which reveals a certain mechanism of its promoting angiogenesis.

BACKGROUND: Strontium-doped calcium polyphosphate (SCPP), as a new repairing material, has been demonstrated to have favorable biocompatibility and biodegradability and some effects on promoting self-angiogenesis. However, the mechanism remains still unknown. OBJECTIVE: Endothelial cells were cultured with SCPP scaffolds in vitro, as well as the cell proliferation and angiogenic factor matrix metalloproteinase-2 (MMP-2) secretion were observed. DESIGN,TIME AND SETTING: A contrast study was performed at the Laboratory of Tissue Engineering of Sichuan University from September 2008 to April 2009. MATERIALS: A series of calcium polyphosphate (CPP) respectively containing 1 %, 2%, 5%, 8%, and 10% Sr~(2+) were prepared.METHODS: ① Materials were plated on 24-well culture plate,and endothelial cell suspension (300 μL) were seeded on 24-well culture plate at the concentration of 3×10~7/L and cultured with 200 uL RPMI1640 culture media. Endothelial cell proliferation was observed using MTT method at days 1,3,5, and 7 after culture. ② CPP and 8% SCPP were plated on 24-well culture plate, and endothelial cell suspension (300 uL) was then incubated in 24-well culture plate at the concentration of 1x10~8/L and cultured with 600 uL RPMI1640 culture media. The morphology of endothelial cells was observed by scanning electron microscopy (SEM) at day 5 after culture.③ Endothelial cells were co-cultured with SCPP of various Sr~(2+) contents for 5 days. After confluence, cells were centrifuged to obtain supernatant. Angiogenic factor MMP-2 secretion was evaluated by ELISA assay.MAIN OUTCOME MEASURES: The proliferation and morphology of endothelial cells on SCPP and CPP were observed. The amount of endothelial cells-derived MMP-2 protein secretion was detected. RESULTS: MTT method demonstrated that the proliferation of endothelial cells on the 8% SCPP scaffold showed a higher level compared to CPP, and other SCPP groups. Scanning electron microscope results suggested that endothelial cells on 8% SCPP had a better growth status and biological activity. ELISA results indicated that angiogenic factor MMP-2 expression on the SCPP was promoted compared with that of CPP, and 8% SCPP showed the highest expression (P < 0.05). CONCLUSION: SCPP has good compatibility with endothelial cells,promoting angiogenesis and enhancing the angiogenic factor MMP-2 expression.

Objective To establish a mouse model of H.pylori infection, and to evaluate the chronic pathological changes in the gastric mucosa associated with H.pylori infection.Methods 34 male 5~6-week old SPF C57BL/6 mice were used in this study.The mice were intragastrically administrated with a suspension of H.pylori SS1 strain.Two weeks after infection, rapid urease test and PCR were performed to confirm the H.pylori infection.Successfully infected mice were randomly divided into 3 groups including the control group, 6-week and 12-week infected groups.Samples of gastric mucosa were taken for pathological analysis using HE and borax methylene blue staining.The contents of myeloperoxidase (MPO), superoxide dismutase (SOD), malondialdehyde (MDA) and catalase (CAT) in the gastric tissues were detected by biochemistry, and the expression levels of COX-2, iNOS, TNF-α and IL-1β were examined by RT-qPCR.Results Compared with the control group, H.pylori colonization was observed in the gastric mucosa of the 6-week and 12-week infected mice, with chronic inflammatory cell infiltration, glandular atrophy and intestinal metaplasia to varying extents.The contents of CAT and SOD were significantly decreased, while the levels of MPO and malonaldehyde MDA, and the expression levels of COX-2,iNOS,TNF-α and IL-1β were significantly increased (P<0.05 or P<0.01).Conclusions Intragastric administration with H.pylori in C57BL/6 mice can be successfully used to generate the bacterial colonization, leading to chronic inflammatory cell infiltration, enhanced oxidative stress, and up-regulated expression of proinflammatory genes in the gastric glandular tissues at 6 and 12 weeks after inoculation.However, the inflammatory changes are more extensive in the mice at 12 weeks after infection, with glandular atrophy and intestinal metaplasia.