Objective To investigate the expression of matrix metalloproteinase-2 (MMP-2) mRNA in lungs of rabbits with acute lung injury (ALI) induced by endotoxin (ET) and the effects of meloxicam in its treatment, and to explore the protective mechanism of meloxicam. Methods Twenty four Japanese flap-eared white rabbits were randomly assigned to three groups: control group, ET-challenge group and meloxicam-treatment group. There were eight rabbits in each group. ALI model of rabbits was replicated with intravenous ET injection (700?g/kg weight), and meloxicam (2.5mg/kg weight) was intravenously given afber ET challenge in the treatment group. During the experiment, arterial blood gases, lung wet/dry (W/D), lung water content, the pathologic changes in lung, the expressions of MMP-2 mRNA in the lungs were determined. Results Compared with control group, oxygen saturation index (PaO_2/FiO_2) was significantly decreased and pulmonary edema, hemorrhage, extensive inflammatory cells infiltration were observed in ET group. The lung wet/dry (W/D), lung water content and the expressions of MMP-2 mRNA were significantly higher in ET group than those in control group. In meloxicam-treatment group, PaO_2/FiO_2 remained in normal range, and the lung wet/dry (W/D), the lung water content and the expressions of MMP-2 mRNA were significantly lower than those in ET group. The pathologic changes in lung tissues were not severe in group C. Conclusion The up-regulation of MMP-2 mRNA expression was observed in ALI animal model. Meloxicam could down-regulate MMP-2 mRNA expression, producing protective effects on ALI induced by ET in rabbits.

BACKGROUND:Patients with spinal metastases may appear different degrees of pain and abnormal spinal stability, and can be treated with percutaneous baloon kyphoplasty combined with bone cement implantation. OBJECTIVE:To explore the effect of percutaneous baloon kyphoplasty with bone cement implantation on spinal stability and pain in patients with spinal metastases. METHODS:Twenty-three patients with metastatic spinal tumor were treated with percutaneous baloon dilatation kyphoplasty with polymethyl methacrylate bone cement. There were 10 females and 13 males, and their age ranged from 23 to 71 years. The visual analogue scale score, anterior and posterior edge height of vertebral body, quality of life, and motor function score of patients were observed before and after treatment. RESULTS AND CONCLUSION: Compared with before treatment, the visual analogue scale score and motor function score were significantly decreased, while anterior and posterior edge height of vertebral body were significantly increased in 23 patients at 24 hours after treatment (P < 0.05). After 12 months of folow-up, no case appeared to have spinal nerve root damage, serious adverse reactions and bone cement embolism. The patient'squality of life was significantly improved, compared with before treatment (P < 0.05). Experimental findings indicate that percutaneous baloon dilatation kyphoplasty with polymethyl methacrylate bone cement can significantly improve the spinal stability, relieve the degree of pain, and exert exact effects in treatment of spinal metastases.

Objective To observe the pulmonary pathologic changes of endotoxin (ET)-induced acute lung injury (ALI) in rabbits and the potential protective effects of fructose-1,6-diphosphate (FDP) on the ET-induced ALI of rabbits. Methods 24 flap-eared albation rabbits were randomly assigned to 3 groups, 8 for each, as follows: control group (group A), ET-treated group (group B) and combination group (treated by ET and FDP, group C). ALI was induced by injection of ET at one time. Group A was only injected with placebo, normal saline. ET was given to the rest groups. In group C, FDP was given as an intervening measure after rabbits injured. Rabbits were sacrificed at 6h time point. The pulmonary pathologic changes were observed. Some markers of pulmonary tissues, including the content of lipid peroxide (LPO), thromboxane B2 (TXB2), 6-keto-prostaglandin F1α(6-keto-PGF1α), interleukin-13 (IL-13) and the activity of superoxide dismutase (SOD), were observed. Results Compared with group A, the contents of LPO and TXB2 of group B showed significant increase (P＜0.05, P＜0.01), the SOD activity of group B weakened obviously (P＜0.01), the contents of 6-keto-PGF1α and IL-13 showed no statistical differences. The LPO content and the SOD activity of group C were similar to those of group A, the contents of TXB2, 6-keto-PGF1α and IL-13 of group C were much higher than those of group A (P＜0.01). Estimated by light microscope and electron microscope, the structure of lung tissue of group A is basically normal, the pathologic injuries of lung tissue of group B were much more severer and that of group C were slighter. Conclusion In the progress of ET-induced ALI, the oxidative injury, imbalance of TXB2/6-keto-PGF1α ratio and the secretion deficiency of protective cytokines play important role in inducing pathologic injuries of lung tissues. FDP can inhibit oxidative injury, ameliorate TXB2/6-keto-PGF1α balance and promote the secretion of protective cytokines, which, in turn, can protect rabbits from ET-induced ALI to some extent.

Objective:To detect the influence of Ipas, and construct better Shigella vaccines.Methods: Guniea pigs were immunized with Ipa + and Ipa - bivalent Shigella vaccines strains by eyes mucosal immunization, and challenged with willed strains. The specific antibodies in tears were measured by BA ELISA assays after immunization and challenge. Results: The levels of specific local specific antibodies of immunized groups increased significantly compared to the control group and Ipa + group compared to Ipa - group after immunization.Conclusion: The expression of invasive plasmid antigens(Ipa) can enhance the levels of specific local antibodies. The two Shigella vaccines have good immunogenicity.

BACKGROUND:Studies concerning bone marrow mesenchymal stem cells (BMSCs) cultured by umbilical cord serum in vitro were hot topic to avoid the heterogenous serum rejection during BMSC culture and improve culture efficiency of BMSCs.OBJECTIVE:To compare biological feature of BMSCs by the umbilical cord serum and adult autoserum in vitro.DESIGN,TIME AND SETTING:The opening experiment was performed at the Laboratory of Cell Biology,Xiangya Second Hospital,Central South University,China from October 2006 to June 2008.MATERIALS:Sixteen bone marrow samples were provided by Xiangya Second Hospital,Central South University and Xiangtan Central Hospital of Hunan Province.METHODS:BMSCs were obtained from 16 fresh adult bone marrow by gradient centrifugation with Ficoll Paque,cultured with DMEM/F12 with 10% umbilical cord blood serum and 10% adult autoserum.The fourth passage was used in this study.The surface antigens of BMSCs were detected by flow cytometry.BMSCs could differentiate into ostenblasts and adipocytes under culture in conditioned medium for osteogeneais and adipogenesis and the differentiated BMSCs were identified by morphological observation,immunophenotype and immunochemical staining.MAIN OUTCOME MEASURES:The following parameters were measured:morphological observation;cell cycle and cell count;identification of surface antigen;observation of osteoblasts and adipocytea induced from BMSCs by immunochemical staining.RESULTS:The quantity and velocity of BMSCs in umbilical curd blood serum was better than those in adult autoserum (P ＜ 0.05)Only few cells were proliferating in both medium,BMSCs in S phase in umbilical cord blood serum was more than those in adult autoserum (P ＜ 0.05).The positive rate of CD29,CD73 and CD105 on BMSCs in umbilical cord serum (above 90%) was higher than those in adult autoserum (P ＜ 0.05),and the positive rates of CD31,CD34 and CD45 were lower than 2%,and the positive rate of CD31 was lower than those in adult autoserum (P ＜ 0.05).The positive rate of BMSCs differentiated into osteoblasts and adipocytes in umbilical cord blood serum was also higher than these in adult autoserum (P ＜ 0.05).CONCLUSION:Purity and differentiated potency of BMSCs in umbilical cord blood serum are better than those in the adult autoserum in vitro.The umbilical cord serum is more adapt to clinical application.

Objective:To prepare compound fluconazole gel and to establish a method for its quality control. Methods: Flucon-azole and baicalin were the main ingredients in the gel, and their contents were determined by HPLC. Moreover, the drug release in vitro, irritation and stability of the gel were tested. Results:The fluconazole and baicalin had a good linear relationship within the range of 12. 5-150. 0 μg·ml-1 and 25. 0-175. 0 μg·ml-1, respectively. The average recovery rate was 99. 45%(RSD=0. 40%, n=9) and 99. 31%(RSD=0. 31%, n=9), respectively. The in vitro drug release of the gel was accordance with the first order release e-quation, and the cumulative release rate of fluconazole and baicalin in 12 h was 87. 6% and 80. 03%, respectively. The gel was stable without irritation. Conclusion:The formula, preparation technology and stability of compound fluconazole gel are promising and the quality standard is controllable.

The effect of anisodine on brain monoamine was studied in 30 rats. The brain monoamine levels of caudate nucleus, hippocampus, diencephalon, brain stem and cerebral cortex were estimated by fluoro-metric method in drug-treated rats and saline-treated controls. Only the NE level of the brain stem was significantly increased in anisodine-treated animals 48 hours after injection. There were no significant differences in the dopamine values between anisodine-treated rats and saline-treated controls, while the 5HT and 5HIAA levels were significantly increased in anisodine-treated animals. The relation of brain monoamines to learning and memory is discussed. The increased 5HT and 5HIAA levels caused by anisodine may play a role in the impairment of memory.

Objective To set up a technical platform of reverse genetics based on the 8 plasmid.virus rescue system of cold-adapted influenza virus strain. Methods The cold-adapted, temperature sensitive, live attenuated influenza virus strain A/AnnArbor/6/60(H2N2)was chosen as the master donor virus(MDV)for rescue research,and its six internal gene fragments PB2,PB1,PA,NP,M and NS were artificially synthesized. Meanwhile, five amino acid mutations have been introduced as tags. Six fragments were ligated with modified pAD3000 for the construction of rescue plasmid. Six transcription/expression plasmids(pMDV-A-PB2,pMDV-A-PB1,pMDV-A-PA,pMDV-A-NP,pMDV-A-M,and pMDV-A-NS)were obtained, and their sequences were accurate. Results The reassorted virus named as rMDV-A contains HA and NA gene segments derived from PR8 strain along with six gene segments,PB2,PB1,PA,NP,M and NS,from MDV. The COS-1 cells were co-transfected with eight recombinant plasmids. The results showed that a cold-adapted, attenuated reassortant influenza A virus with hemagglutination activity was rescued successfullv bv"6+2" combination of MDV and PR8, and the allanotoic fluid of the injected eggs gave a posigenes of A/AA/6/60 used as backbone has provided experimental materials for further research on the gene function and novel vaccine candidate of cold-adapted, attenuated human influenza virus.

Six gene segments,PB1,PB2,PA,NP,M and NS,were fully synthesized which derived from the master donor virus (MDV),cold-adapted(ca),temperature sensitive(ts),live attenuated influenza virus strain A/Ann Arbor/6/60(MDV-A).Meanwhile,five amino acid substitutions (PB1-391E,58lG,661T,PB2-265S,NP-34G) were artificially altered by human intervention.HA and NA fragments derived from the 2006-2007 circulating strain A/New Caledonia/20/99 (H1N1).Eight fragments were ligated with modified pAD3000 for rescue plasmid construction.Eiight transcription/expression plasmids were named as pMDV-A-PB2,pMDV-A-PB1,pMDV-A-PA,pMDV-A-NP,pMDV-A-M,pMDV-A-NS,pMDV-A-HA,pMDV-A-NA,respectively.The COS-l cells were co-transfected with eight plasmids representing 6 internal viral backbone of the strain A/AA/6/60 and two plasmids containing the CDNA of the HA and NA segments of the strain A/New Caledonia/20/99 (H1N1),the results showed that cold-adapted,attenuated reassortant influenza A virus Was rescued successfully.Titers of a reassorted influenza A virus in embryonated chicken eggs mnged from 1:29to l:210.The rescue system of six intemal genes used as backbone opens the way for further research on gene function and neotype vaccine candidate of cold-adapted,live attenuated human influenza virus.

Six gene segments, PB1, PB2,PA, NP, M and NS, were fully synthesized which derived from the master donor virus(MDV), cold-adapted(ca),temperature sensitive(ts), live attenuated influenza virus strain A/Ann Arbor/6/60(MDV-A). Meanwhile, five amino acid substitutions (PB1-391E, 581G, 661T, PB2-265S, NP-34G) were artificially altered by human intervention. HA and NA fragments derived from the 2006～2007 circulating strain A/New Caledonia/20/99 (H1N1). Eight fragments were ligated with modified pAD3000 for rescue plasmid construction. Eight transcription/expression plasmids were named as pMDV-A-PB2, pMDV-A-PB1, pMDV-A-PA, pMDV-A-NP, pMDV-A-M, pMDV-A-NS, pMDV-A-HA, pMDV-A-NA, respectively. The COS-1 cells were co-transfected with eight plasmids representing 6 internal viral backbone of the strain A/AA/6/60 and two plasmids containing the cDNA of the HA and NA segments of the strain A/New Caledonia/20/99 (H1N1), the results showed that cold-adapted, attenuated reassortant influenza A virus was rescued successfully. Titers of a reassorted influenza A virus in embryonated chicken eggs ranged from 1∶29 to 1∶210. The rescue system of six internal genes used as backbone opens the way for further research on gene function and neotype vaccine candidate of cold-adapted, live attenuated human influenza virus.