Objective To investigate the risk factors associated with surgical site infection (SSI) in patients with breast cancer,and provide the basis for preventing SSI.Methods Case-control study was used to investigate the clinical data of 630 patients of breast cancer retrospectively by single factor and multiple factors regressive analysis.Results The incidence rate of SSI was 3.65％ (23/630),the main pathogenic bacteria was staphylococcus aureus (14/20),single factor regressive analysis showed that granulocytopenia associated with chemotherapy,diabetes mellitus,time of drainage were the risk factors of SSI (P ＜ 0.05).Multiple factors regressive analysis showed that granulocytopenia associated with chemotherapy,time of drainage were the independent risk factors of SSI (P ＜ 0.05).Conclusion Granulocytopenia associated with chemotherapy,time of drainage are the independent risk factors of SSI and comprehensive preventive measures should be taken to control the incidence rate of SSI.

Objective To explore the clinical effect of treatment for progressive multiple sclerosis(PMS)by autologous peripheral blood stem cell transplantation(APBSCT).Methods Between September 2001 and July 2004,thirteen patients with PMS were treated with APBSCT in Xuanwu Hospital.Peripheral blood stem cells (PBSCs)were mobilized with G-CSF alone.The stem cell transplants were pured of lymphocytes in 7 patients.The high dose regimen was BEAM conditioning regimen (Carmustine,Etoposide,Cytosine Arabinoside,and Melphalan).The median follow-up time was 22 (3～36)months.Results The result showed that the mean EDSS rating score of patients in 18 months after APBSCT was significantly decreased[(4.05?0.66) vs(6.00?0.30),P

Objective To investigate the animal model of acute lung injury (ALI) induced by caudal vein injection of trypsin in rats and to evaluate the model.Methods The model of lung injury was established by caudal vein injection of trypsin in rats.The rats were killed at the time point of 3 h,6 h,12 h,24 h,and 24 h and then the pathological changes of structure of lungs,peripheral blood neutrophil count,arterial blood gas analysis and lung wet/dry (W/D) weight ratio in rats were measured and observed.Results The results of hematoxylin eosin (HE) staining showed that there was no obvious pathological changes in lung tissues of the control group,while alveolar and pulmonary septal edema,thickening,and a large number of inflammatory cells infiltration in the lung tissues of the model group.Compared to the control group,the peripheral blood neutrophil counts,W/D and PaCO2 were significantly increased,PaO2 was significantly decreased (P ＜0.01).There was significant differences in the number of peripheral blood neutrophils,PaCO2,W/D and PaO2 between the model groups (P ＜ 0.01).Conclusions The rat model of ALI induced by trypsin can successfully simulate the lung damage caused by the release of a large number of trypsin when severe acute pancreatitis occurred.

BACKGROUND:The cell-sheet technology, based on a temperature-responsive culture, has been drawing more and more attention;however, the temperature-responsive culture dish is quite expensive. Therefore, it is imperative to develop a substitutive technique.OBJECTIVE: To study the feasibility of cell-sheet ulturing using common culture dish, and investigate the chondrogenesis of the cell sheet. METHODS: A piece of nasal septal cartilage was adopted from a patient with deviation of nasal septum to extract primary chondrocytes that were then cultured and amplified. The passage 3 chondrocytes were used to construct ell sheets. Monolayer cell sheet was formed by intensive culturing and allowing the extracellular matrix secretion. Bilayer cell sheet was constructed by seeding passage 2 chondrocytes on the monolayer cell sheet. The cell sheets were harvested using cell scraper, their properties were investigated prior to plantation into nude mice to construct the tissue-engineered cartilage. RESULTS AND CONCLUSION: Both bilayer and monolayer cell sheets with soft tremellose structures showed no significant difference through naked eyes. The newly harvested cell sheets appeared to have good fluidity and gelation. Eight weeks after mplantation into the nude mice, mature cartilage blocks were obtained. Histologically, the cell sheets were thin films composed by layered chondrocytes and extracellular matrix. Glycosaminoglycan formation and type Ⅱ collagen expressions were observed in the cell sheets cultured in vitro. The explanted samples exhibited ature cartilaginous tissue at 8 weeks after implantation. Biochemical analysis showed that the DNA contents of the neocartilages were higher than those of native human costal cartilage, while the contents of glycosaminoglycan and hydroxyproline were similar to native human nasal septal cartilage. To conclude, the hondrocyte cell sheets are likely to be constructed and harvested successfully using common culture dish, and the cell sheets exhibit favourable chondrogenesis.

Objective]To study the feasibility of Cartilage engineering using fibrin gel and chondrocyte cell sheets.[Methods]rabbit auricular chondrocytes were isolated and cultured to form cell sheets in flasks. The cell sheets were harvested using cell scrapers,and cut into fragments. The two precursor solutions of Fibrin gel were used to suspend the cell sheet fragments and isolated chondrocytes,and then added into the wells of a 48-well plate to form Gelatinous chondroid disc constructs. After in vitro culture, the constructs were implanted into nude mice. After 8 weeks,the constructs were harvested,and the specimens were evaluated using grossly observing, histological and immunohistochemical observation. [Results]Mature cartilage discs were obtained. The histomorphology of the explanted discs appeared non-uniform cartilaginous tissue comprise of regenerated cartilage islands with different size and irregular shape. Immunohistochemistry staining demonstrated that type II collagen highly expressed in the ECM of the cartilage islands. In 1 of the 8 discs,partial ossification was observed.[Conclusion]Fibrin gel is a favourable carrier. Artificial cartilage with stereochemical structure was constructed via combining the fibrin gel and chondrocyte cell sheets.

Objective To study the effect of osgentide (OST) on proliferation of mouse preosteoblast MC3T3-E1 under simulated microgravity ( SMG ) .Methods Under normal conditions , cell proliferation was evaluated by MTT assay to screen an OST compound of an effective concentration after MC 3T3-E1 cells were treated with series OSTs .Furthermore, cell proliferation and cell cycle distribution of MC 3T3-E1 cells were analyzed after treatment with 1 nmol/L OST5 by MTT assay and by flow cytometry ( FCM) scanning under SMG .Results Under normal conditions , 1 nmol/L OST5 was able to significantly promote the proliferation of MC3T3-E1 cells (P<0.01).Under SMG, proliferation of MC3T3-E1 cells was significantly inhibited and more cells entered G 1 than under normal conditions (CN).The proportion of S phase of MC3T3-E1 cells after treatment with 1 nmol/L OST5 ( OST-SMG) for 3 d was higher than that of untreated MC 3T3-E1 cells under SMG,suggesting that OST5 could promote DNA synthesis ( P<0.05 ) .Conclusion OST5 facilitates the proliferation of MC3T3-E1 cells under SMG, which provides a basis for the use of OST5 in the prevention and treatment of bone loss relat-ed to microgravity .

Objective To investigate the effect of simulated microgravity on growth , morphology, protein expression and virulence gene expression of Klebsiella pneumoniae (KPN).Methods KPN was divided into simulated microgravity group and control group in the experiment .The former group was in the ambient of simulated microgravity in a clinostat .The bacterial growth curves , morphologyical changes in electron microscopy , and protein expression were detected by SELDI-TOF-MS, and the expression of 4 virulence genes(ureA,wabG,uge and fimH) by real-time fluorescence quantitative PCR (RT-PCR) in both groups.Results Compared with the control group , the growth of KPN under simulated microgravity was accelerated , and the total bacterial count increased in microgravity group .The bacterial morphology in microgravity group was changed under scanning electron microscopy (SEM), and thinner and longer bacteria were increased .The transmission electron microscopy ( TEM) analysis revealed increase in cytoplasmic granular substance in microgravity group .Proteome analysis showed that the expression of 18 proteins was changed , half of which up-regulated and the rest were down-regula-ted.Those 18 proteins were searched in the protein library .And 21 proteins of a similar molecular mass were retrieved ,13 of which,proteins with known functions ,were closely related to bacterial life activities .RT-PCR results showed that four virulence genes of KPN were down-regulated.Conclusion Upon exposure to simulated microgravity , the growth and repro-duction of KPN are accelerated and enhanced .The bacterial morphology is changed .The strain′s protein expression and four virulence genes expressionare also changed .Therefore,microgravity can change the characteristics of KPN .

BACKGROUND:Research on ethyl pyruvate detection methods is reported rarely, and moreover, literature about reversed-phase high-performance liquid chromatography (RP-HPLC) for detection of ethyl pyruvate is less.
OBJECTIVE:To establish an RP-HPLC method for determination of ethyl pyruvate in ethyl pyruvate-chitosan nanoparticles.
METHODS: The chromatographic analysis was performed on a ZORBAX Eclipse XDB-C18 column (4.6 mm× 150 mm, 5μm) at 25℃, with the mixture of acetonitrile and water (40:60, V/V) as the mobile phase at the flow rate of 1 mL/min. The determination wavelength wasset at 210 nm and the injection volume was 20 μL.
RESULTS AND CONCLUSION: The peak of ethyl pyruvate and the peaks of auxiliary materials and solvent were separated wel. The linear rang of ethyl pyruvate was 1-100 mg/L (r=0.999 6). The relative standard deviation of both the intra-and inter-day precision was less than 3% for low-, moderate-, and high-concentration ethyl pyruvate. The relative standard deviation of reproducibility test and stability test was 1.25% and 1.3%, respectively. Sample average recovery rates were (91.5±1.0)%, (3.5±0.2)%, (94.4±0.4)%, respectively. Encapsulation efficiency of samples were (87.2±0.22)%, (90.5±0.15)%, (91.1±0.17)%, respectively. The relative standard deviation of different sample content were 0.9%, 0.5%, 0.3%, respectively. The RP-HPLC method for determination of ethyl pyruvate is sensitive, accurate and highly specific with wide linear range and high sample average recovery.

BACKGROUND:To seek for ideal scaffold materials is still an important task for cartilage tissue engineering.OBJECTIVE:To investigate the application of the AviteneTM microfibrillar collagen hemostat sponge in cartilage tissue engineering.METHODS:Rabbit auricular cartilage was harvested via surgical operation,and primary chondrocytes were isolated and amplified.Microfibrillar collagen hemostat sponge was cut into small bricks.The passage 2 chondrocytes were suspended and seeded onto the spongy bricks.After 1 week of in vitro culture,the constructs were then implanted into nude mice.After 8 weeks,the specimens were collected and evaluated using gross,histological and immunohistochamical observation.RESULTS AND CONCLUSION:During the cell seeding,the scaffold maintained its dimensions.No shrinkage was observed when the cell suspension was added.There was no considerable change in dimensions during the 1-week in vitro culture and at 8 weeks after implantation in nude mice.At 8 weeks post-implantation,mature cartilage blocks were harvested,which were white,translucent,and flexible.Histologically,the constructs appeared to have typical mature cartilaginous tissues,with robust extracellular matrix secretion,in which the microfibrillar collagen was incompletely degraded.We conclude that the microfibrillar collagen is a favorable scaffold material for cartilage tissue engineering.

Objective To investigate the effects of simulated microgravity by rotary cell culture system (RCCS) on expression profiles of miRNA in mouse fibroblasts L929 cell line.Methods L929 cells were cultured in vitro and divided into simulated microgravity (SMG) group and normal gravity (NG) group according to the simple random method.Samples of two groups were collected on 7th day of culture and the total RNAs were extracted,labeled,and hybridized in sequence.Feature Extraction Software was used to collect the array images and get raw data,which were analyzed by Genespring Software.Differentially expressed miRNAs were identified and then validated by qRT-PCR.Target genes of differentially expressed miRNAs were predicted by the databases of Targetscan and microRNAorg,and the intersections of databases were identified as potential regulatory target genes.Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) analysis were applied to determine the roles of these target genes.Relevant biological functions and/or signaling pathways of the regulated genes especially related with wound healing process were categorized based on their enrichments.Then integration predictions of the miRNA and mRNA expression profiles had been proposed to refine the functional miRNA-mRNA relationships.The miRNA-mRNA functional network and miRNA-mRNA control network were constructed.Results Four miRNA genes were up-regulated significantly including mmumiR-669j,-122-5p,-30a-3p,-6516-3p,among which mmu-miR-669j was up-regulated at 52.84 folds with the greatest significance (P ＜ 0.05).Seventeen miRNA genes were down-regulated significantly including mmu-miR-21a-3p,-miR-28a-5p,-218-5p,-210-3p,-miR-19a-3p,-miR-31-3p,and-miR-19b-3p,among which mmu-miR-28a-5p was down-regulated at 15.47 folds with the greatest significance (P ＜ 0.05).The qRT-PCR showed a high concordance with the microarray results (P ＜ 0.05).Target gene prediction and functional enrichment analysis suggested that a variety of biological processes and signaling pathways involved in wound repair were significantly enriched (P ＜ 0.05).Function network and regulation network of miRNA-mRNA covered all the differentially expressed miRNAs,which suggested that miR-21 a-3p and predicted target gene Smad3 might play an important role in wound healing under microgravity.Conclusions Simulated microgravity by RCCS can significantly affect the expression of stress-related miRNAs in mouse fibroblasts L929.The miRNA target gene prediction and functional enrichment analysis based on gene chip technology may provide theoretical basis for illustrating the mechanism and management of weightlessness stress injury.