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 identify Mallotus panicutus (lam) Muell.Arg and explore the acute toxicity.Methods Pharmacognostic identification and chemical reaction methods were adopted to the identification.The maximum concentration and maximum volume of ethanol extracts of M.panicutus (lam) Muell.Arg were given via intragastric administration in mice to observe the toxic and side effects after 14 days.Results The chemical reaction showed M.panicutus(lam) Muell.Arg concluded the constituents such as flavone and saponin and there was no acute toxicity in mice.After 14 days,the growth rate of mice weight of blank group and drug group were 73.5% and 62.9% (P ＞ 0.05).Conclusion The results could be taken as the reference for analyzing the quality of this drug and guiding the clinical medication.

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: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: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 study the anti-senile and anti-oxidative activities of Forsythia suspense leaves tea(FSLT) in mice. Methods: 36 ICR mice(7 mo age) were randomly divided into three groups: senior control group,ig distilled water ; FSLT1 group,ig FSLT 0.3 g ?kg-1?d-1 ; and FSLT2 group,ig FSLT 0.9 g?kg-1?d-1 .The other 12 ICR mice(young control group) were given ig distilled water. MDA and MAO-B in brain and mitochondria were measured. SOD,MDA and POD in heart, liver index, thymus index and spleen index were determined. Results: Forsythia suspense leaves tea may reduce the level of MDA and MAO-B, and increase the activities of SOD and POD. Conclusion: Forsythia suspense leaves tea may have anti-senile and anti-oxidative activities.

A total of 68 patients with histologically proven endometrial cancer were preoperatively evaluated with a 3.0 T magnetic resonance (MR) unit.Two radiologists interpreted the depth of myometrial invasion on T2-weighted and diffusion-weighted(DW) MR (b =700 s/mm2) imaging.Myometrial tumor spread was classified as superficial (＜ 50％) or deep (≥50％ myometrial thickness).Statistical methods included Kappa statistics for reader agreement,accuracy assessment and receiver operating characteristic analysis for diagnostic performance comparison.For assessing the depth of myometrial invasion,the diagnostic accuracy,sensitivity and specificity were as follows:T2-weighted imaging-reader 1st,61.8％,60.0％ and 65.2％;reader 2nd,66.2％,64.4％ and 69.6％ ; T2-weighted combined DW imaging-reader 1st,86.8％,84.4％and 91.3％ ; reader 2nd,89.7％,86.7％ and 95.7％.Reader agreement was excellent for T2-weighted combined DW imaging (Kappa =0.818).For assessing deep myometrial involvement,ROC analysis showed that the diagnostic accuracy was significantly higher on T2-weighted combined DW imaging than that on T2-weighted imaging.The AUC (area under the curve) values measured by two readers on T2-weighted and T2-weighted combined DW imagings were 0.626,0.879 and 0.670,0.912 respectively.The addition of 3.0T MR diffusion-weighted to T2-weighted imaging can improve the diagnostic performance of MR imaging in the assessment of myometrial invasion.

Objective To investigate the safety , feasibility , and clinical application value of digital sub-traction angiography ( DSA) -guided insertion of totally implantable venous access port ( TIVAP) in patients with digestive system cancer .Methods We retrospectively analyzed the data of 15 digestive system cancer patients who were inserted with TIVAP under DSA guidance between April 2013 and January 2016, recorded the TIVAP-related complications and indwelling time, and investigated the patients’satisfaction about TIVAP.Paired rank sum test was used for the differences in patients’quality of life before and after the insertion of TIVAP.Results Of the 15 digestive system cancer patients , operation success rate of TIVAP insertion was 100%, with the success rate of venipuncture on first attempt being 100%.The incidence of complication was 6.67% ( 1/15 ) , which was manifested as pull feeling at the neck occurring in the eighth month after insertion .The indwelling time of TIVAP was from 2 to 28 months, with a median value of 9 months.Patients’satisfaction rate of TIVAP was 86.67%(13/15).The proportion of patients with a good quality of life was 100%(15/15), statically signifi-cantly higher than that before the insertion [46.67%(7/15), Z=-3.416, P=0.000).Conclusions TIVAP insertion under DSA guidance in digestive system cancer patients is safe and feasible , with few complications and fair patient satisfaction .It may improve the patient ’s quality of life , worthy of clinical application .

Objective To investigate the possibility of perform depression screening with the patient health questionnaire (PHQ-9) in hepatocellular carcinoma patients,and to evaluate the prevalence of depression among these patients.Methods 186 hepatocellular carcinoma patients completed PHQ-9 and Hamilton depression scale (HAMD) from January 2015 to April 2015.60 patients randomly selected from these 186 patients received depression screening again with PHQ-9 in 7 to 14 days after the first survey.Results Cronbach's α coefficient of PHQ-9 was 0.869,the test-retest reliability 0.963,and the range of correlation coefficient of the nine items with the total score of the scale was 0.546-0.752 (P＜0.01).The sensitivity and specificity of PHQ-9 were 84.4％ and 95.7％ respectively.69(37.1％) cases of the 186 patients were detected with depression with PHQ-9,and mild,moderate and major depression were 41 (22.0％),21 (11.3％) and 7(3.8％) respectively.Conclusion PHQ-9 has been shown to have good reliability and validity for screening of depression in hepatocellular carcinoma patients.

Objective Observe the effect and the indications of intermittent short veno-venuous hemofiltration(ISVVH) in severe acute pancreatitis(SAP). Methods APACHE II scores≥14 and fluids imbalance were respectively used to define as the indication of starting hemofiltration and ending hemofiltration. In 39 patients with SAP,19 underwent ISVVH(IS group),and the other 20 patients were not accepted hemofiltration (N group). APACHE II scores, Balthazar CT grades and the plasma levels of procalitonin(PCT), TNF-?, IL-6, IL-8, IL-1ra, IL-2 and IL-10 were observed. Results At admission and 2d after admission, APACHE II scores in IS group and N group were (13.8?3.1)and (17.8?3.2) ( P