BACKGROUND: Some studies have demonstrated that the degradation of extracellular matrix (ECM), which matrix metalloproteinases (MMPs) participates in, plays a key step in the corneal neovascularization (CNV). Tissue factor pathway inhibitor 2 (TFPI-2), a new type serine proteinase inhibitor found recently, can effectively inhibit the activity of MMPs. Whether TFPI-2 gene transfection can influence CNV is unclear.OBJECTIVE: To investigate the effect of TFPI-2 gene transfection on CNV.DESIGN: Randomized controlled experiment.SETTING: Laboratory for Department of Surgery, Wuhan Union Hospital; Central Laboratory, the Affiliated Third Hospital of Sun Yat-sen University.MATERIALS: This study was carried out in the laboratory for Department of Surgery of Wuhan Union Hospital and State Central Laboratory of the Third Hospital Affiliated to Sun Yat-sen University between June 2004 and March 2006. Sixty healthy purebred adult New Zealand rabbits of either gender, weighing 2.5 to 3.0 kg, were involved. Preoperatively, no obvious anterior segment ocular lesion was found by slit-lamp examination. pBos-Cite-neo/TFPl-2 was kindly gifted by Dr. Zhong Ren (Department of Hematology, Union Hospital). Peroxydase blocking agent, nonimmune goat serum,mouse anti-human MMP-1, 2 and 3 monoclonal antibodies, biotin labeled goat-anti-mouse IgG second antibody (Santa cruz Company) were used in this study.METHODS: Experimental intervention: Experimental rabbit models of CNV were created in each group by silver nitrate cautery. Then, the rabbit models were randomized into 3 groups and 20 rabbits for each group. Different reagents were subconjunctivally injected via many points in each group: saline in the group Ⅰ, empty vector in the group Ⅱ, plasmid encoding TFPI-2 in the group Ⅲ. Experimental evaluation: CNV growth was observed under the slit-lamp biomicroscope.The expression of TFPI-2 in each rabbit model was detected by reverse transcription-polymerase chain reaction (RT-PCR) method 2 weeks after modeling; the expression of MMPs in corneal tissue was detected by immunohistochemical method at 3,5,7,9 and 14 days after modeling.TFPI-2 gene expression was significantly higher in the group Ⅲ than in the group Ⅱ and group Ⅰ (P ＜ 0.01); The MMP-1, 2, 3 expressions in the corneal tissue were significantly lower in the group Ⅲ than in the group Ⅱ and group Ⅰ,respectively, especially MMP-1, 3.

Objective To investigate the effect of different doses of microbe?derived antioxidant on sleep and antioxidant ability in mice. Methods Sixty male Kunming mice with similar body weight were randomly divided into 4 groups. The control group received normal saline, and the experimental groups received microbe?derived antioxidant in a dose of 0?5 g/kg bw, 1?0 g/kg bw or 1?5 g/kg bw once per day, respectively. The experiment period was 30 days. At the end of experiment, the mice of each group were intraperitoneally injected sodium pentobarbital to induce sleep. The mice fall sleep was judged by righting reflex. After the test of sleep, blood was taken for detection of serum antioxidant ability. Results Compared with the low dose and high dose groups, the middle dose group showed a significantly prolonged sodium pentobarbital?induced sleeping time (P<0?05). Compared with the control group, low and high dose groups, the middle dose group had highly significantly increased GSH?Px activity ( P <0?01 ) and significantly increased content of SOD. Under these conditions, the middle dose group reduced both the contents of MDA and 8?ISO?PGF2α ( P<0?05 ) compared with the control group. Conclusions Our results suggest that microbe?derived antioxidant exerts effect on sleep and antioxidant ability in rats. Supplement of 1?0 g/kg bw/d shows the most significant effects.

BACKGROUND:Engineered hepatic tissue is considered a promising strategy for healing acute liver failure. But, there are series of hindrances in the construction of engineered hepatic tissues, including acquisition of vital hepatocytes, choice of scaffolds and culture system, and nutrition supply. OBJECTIVE:To construct three kinds of engineered hepatic tissues in hope to screen the optimal one for transplantation in acute liver failure. METHODS:After purification, amplification, bone marrow mesenchymal stem cells were induced to differentiate into hepatocyte-like cells which were co-cultured with acellular amniotic membrane, DACRON PATCH cardiovascular surgical patch, biological surgical patch, respectively to construct three kinds of engineered hepatic tissues. After 3 days of culture, morphological and functional detections were performed. RESULTS AND CONCLUSION:Rat bone marrow mesenchymal stem cells with higher purity were successful y harvested by using density gradient centrifugation and adherent methods, and then the cells were differentiated into hepatocyte-like cells. In the three kinds of engineered hepatic tissues, hepatocyte-like cells were found to be combined with the biological surgical patch to the maximum extent, and their combination exhibited stronger ability of urea synthesis and albumin secretion, which provides experimental basis for treatment of acute liver failure.

BACKGROUND:Previous studies have suggested that achaete-scute homology 1 (ASCL1) plays a key role in the neuronal commitment. Therefore, somatic cels may directly differentiate into neurons by gene transfection ofASCL1, which wil provide new therapeutic strategies for optic nerve regeneration. OBJECTIVE:To construct a recombinant adenovirus vector expressing ratASCL1 gene for further research ofASCL1 gene function. METHODS:The ratASCL1 gene and advenovirus shuttle plasmid (pYr-adshuttle-4) which contained enhanced green fluorescent protein (EGFP) reporter gene were cleaved by restriction endonucleaseXhoI andEcoR I. The target gene fragments were connected together to generate a recombinant plasmid pYr-ads-4-rat-ASCL1 and then transfected into E.coliDH5α. The plasmid was confirmed to be constructed as expectation by enzyme digestion and sequence reaction. The plasmid pYr-ads-4-rat-ASCL1 and pAd/PL-DEST were reconstructed by homologous recombination processes to obtain rat ASCL1 recombinant adenovirus vector. The plasmid pYrAd-rASCL1 was linearized byPac I and subsequently transfected into HEK293 cels for packaging and amplification. RatASCL1 gene in the recombinant adenoviruses were identified by PCR. Virus titer was determined by tissue culture infectious dose 50. Infection efficiency was monitored by EGFP expression. RESULTS AND CONCLUSION:Restriction endonuclease digestion and DNA sequencing showed that the recombinant adenovirus vector pAd-rat-ASCL1 was constructed correctly. The positive amplification bands of 862 bp could be seen in PCR analysis. The virus titer reached 2×1010 pfu/mL. Infection efficiency of recombinant adenovirus in HEK293 cels was more than 80%. The results indicate that the recombinant the adenovirus vector containingASCL1 with high titer and infection efficiency has been successfuly constructed, which can be helpful for further research of the function and clinical application ofASCL1 gene for optic nerve regeneration.

BACKGROUND:The degradation of extracellular matrix, which is mediated by matrix metal oproteinases, plays a crucial role in the corneal neovascularization. Tissue factor pathway inhibitor 2, a new type serine proteinase inhibitor, can effectively inhibit the activity of matrix metal oproteinases.
OBJECTIVE:To elucidate the effect of tissue factor pathway inhibitor 2 on the expressions of matrix metal oproteinases in keratocytes in vitro.
METHODS:Rabbit keratocytes were primarily cultured and subcultured in vitro. Plasmid vector pBos-Cite-neo/TFPI-2 was transfected into keratocytes with Lipofectamine 2000. The positive cells were selected using G418.
RESULTS AND CONCLUSION:The results of reverse transcription-polymerase chain reaction, western blot analysis and gelatinase zymography analysis showed that, expression of mRNA and protein of tissue factor pathway inhibitor 2 was upregulated in the transfected keratocytes (P<0.05), while activity of matrix metal oproteinase 1 and 2 was significantly decreased (P<0.05). Experimental findings indicate that that tissue factor pathway inhibitor 2 strongly inhibits the activity of matrix metal oproteinase 1 and 2 in keratocytes.

Objective To evaluate the standing bending, traction and fulcrum bending radiographies in the prediction of scoliosis correction. Methods 27 patients with thoracic scoliosis with an average of 16.4 years old were selected for the evaluation. All the patients were treated with the posterior correction and bone grafting fusion, and fixed with the third generation system of the segmental vertebral fixation, such as TSRH, CD, or CD+Horizon. Measurement of the Cobbs angle was carried out in the preoperative posterior anterior radiographies with the natural position, standing bending, traction and fulcrum bending and in the postoperative posterior anterior radiographies. Results Mean of Cobbs angle at the thoracic curve after operation was 31 degrees, which decreased significantly in comparison with the preoperative data. Average correction rate was 51.6%. The postoperative Cobbs angle at the thoracic curve was positively correlated with the preoperative Cobbs angle in the standing bending, traction and fulcrum bending radiographies. It was significantly different from the preoperative Cobbs angle in the standing bending and traction radiographies, while no difference was seen with the fulcrum bending radiography. Conclusion The fulcrum radiography can be used to evaluate the flexibility of the thoracic scoliosis, and is better than the standing bending and traction radiographies. The three methods should be used together for the analysis of the scoliosis to predict the result of the scoliosis correction.

ObjectiveTo analyze the prognosis of the new occurred proximal thoracic(PT) curve after posterior correction surgery in adolescent idiopathic scoliosis (AIS) patients.MethodsThe radiographs of 21 AIS patients,who had a primary main thoracic (MT) curve and suffered from a new PT curve after posterior correction surgery,were retrospectively reviewed.All patients were followed up for an average of 19.8 months.Imaging parameters including coronal Cobb angle,T1 tilt and radiographic shoulder height,were measured and analyzed.ResultsThe average coronal Cobb angle of the MT curve was 54.3° preoperatively,14.1° at the final follow-up,with an average correction rate of 74.6％.Before operation,5 patients had negative T1 tilt and 16 without T1 tilt.After operation,all the 21 patients had positive T1 tilt and new PT curves;the average T1 tilt was 6.0° and the average coronal Cobb angle of the PT curve was 16.0°.At the final follow-up,the average T1 tilt and coronal Cobb angle of the PT curve were 4.0° and 13.2°,respectively; eighteen patients still had positive T1 tilt and 17 patients still had PT curves larger than 10°.As for the radiographic shoulder height,5 patients had equal shoulders and 16 patients had right shoulder elevation before operation; after operation,19 patients had left shoulder elevation and 6 patients had unbalanced shoulders; at the final follow-up,14 patients still had left shoulder elevation and 5 patients had unbalanced shoulders.At the final follow-up,T1 tilt and coronal Cobb angle of the PT curve were better than those postoperatively immediately,while the incidences of the new PT curve,left shoulder elevation,unbalanced shoulders were all not significantly different from those postoperatively immediately.ConclusionThe new occurred PT curves after posterior correction surgery in AIS patients are difficult to be compensated and could cause shoulder imbalance,thus great attention should be paid to prevent the occurrence of the PT curve.

Objective To study the effect of single short (SS) veno-venous hemofiltration and intermittent short (IS) veno-venous hemofiltration on imbalance of pro- and anti-inflammatory cytokines in severe acute pancreatitis (SAP) patients. Methods There were 17 and 21 SAP patients indicated for hemofiltration respectively enrolled for (SS) veno-venous hemofiltration (SS group) and IS veno-venous hemofiltration (IS group).Twenty SAP patients admitted and indicated for hemofiltration but not receiving the management during the same period served as control. Plasma levels of TNF-?,IL-6,IL-8,and IL-10 were measured. APACHE Ⅱ scores and fluid balance were used to evaluate patients′ condition. Results APACHE Ⅱ scores in both SS and IS groups declined significantly at day one after admission(P

Objective To investigate the effect of follow-up frequency on the dialysis quality of patients undergoing continuous ambulatory peritoneal dialysis (CAPD). Methods 298 CAPD pa-tients were selected for retrospective analysis from December 2005 to April 2007. All patients were di-vided into two groups according to different follow-up frequency: group A (shorter than 3 months),group B (longer than 3 months). The dialysis quality of the two groups was compared. Results The levels of hemoglobin, albumin and transferrin of group A were (112.19±20.62)mmol/L, (40.45±4.50) retool/L, (2.43±0.29) mmol/L,which were significantly higher than those of group B, (99.63±20.69) mmol/L, (38.01±5.02)mmol/L,(2.29±0.36) mmol/L (P＜0.05). In addition, edema level, life self-care,work capacity, median duration of dialysis, education level and address in group A were significantly different from those of group B (P ＜ 0.05). Conclusion Shortening follow-up frequency plays an im-portsnt role in improving the dialysis quality of CAPD patients.

Objective To evaluate local moderate hypothermia technology on acute brain hemorrhage cases.Methods 100 patients of cerebral hemorrhage were randomly divided into two groups(moderate hypothermia groupand control group). The moderate hypothermia group was 50 cases treated with moderate hypothermia after routine treatment,the control group was 50 cases with routine treatment. The two groups were assessed after treatment for 21 days. Results The group treated via moderate hypothermia therapy showed the nerve function damages relatively lower than the control group(P < 0.05), and the surviving probability and curative ratio were significant higher than the control group(P< 0.05 ). Conclusion The local moderate hypothermia therapy for acute cerebral hemorrhage patient in clinical can significantly improve the never cell function and reduce mortality and disablement.