ObjectiveTo investigate the mechanism by which Notoginsenoside R1 （NGR1） ameliorates hypoxia/reoxygenation （H/R）-induced injury in AC16 human cardiomyocyte cell lines through the regulation of mitophagy. MethodsCommon genes linked to hypoxia/reoxygenation injury and mitophagy were identified by intersecting data from GeneCards and MitoCarta databases. AC16 cell viability was assessed via CCK-8 assay under varying NGR1 concentrations （0， 6.25， 12.5， 25， 50， 100， 200， 300， 400， 500 μmol/L）. AC16 cells were divided into the following groups： control group （Control）， model group （H/R）， and treatment groups （H/R + NGR1 at 100， 200 and 300 μmol/L）. Mitochondrial membrane potential （ΔΨm） was measured using 5，5'，6，6'-tetrachloro-1，1'，3，3'-tetraethylbenzimidazolylcarbocyanine iodide （JC-1） staining. Transcriptional levels of mitophagy-related genes （Parkin， Pink1， P62） were quantified by reverse transcription-quantitative PCR （RT-qPCR）. Protein expression of mitophagy-related markers （Parkin， Pink1， P62， and LC3BⅡ） was evaluated via Western blot analysis. Mitochondrial ultrastructure was visualized by transmission electron microscopy （TEM）. ResultsCompared to the control group， cell viability in the H/R group significantly decreased （P<0.01）. Treatment with NGR1 at concentrations above 100 μmol/L significantly enhanced the cell viability of AC16 cells compared to the H/R group （P<0.01）. H/R induced a significant decrease in mitochondrial membrane potential （P<0.01）， which was restored by NGR1 treatment （P<0.01）. The mRNA levels of Parkin， Pink1， and P62 in the H/R group were upregulated compared to the control group （P<0.05）， while NGR1 intervention downregulated their expression （P<0.05）. Protein expression levels of Parkin， Pink1， and LC3BⅡ in the H/R group significantly increased， while P62 expression decreased compared to the control group （P<0.01）. In contrast， different doses of NGR1 treatment significantly reduced the expression of Parkin， Pink1， and LC3BⅡ while increasing P62 expression （P<0.05）. TEM revealed that the mitochondrial structure in the H/R group was severely disrupted， with fragmented and disorganized cristae， which was alleviated by NGR1. ConclusionNGR1 ameliorates H/R-induced AC16 cell injury， and its mechanism may be associated with modulating the Pink1/Parkin pathway to suppress excessive mitophagy.

Objective To establish a mouse model for human esophageal cancer.Methods Human PBL were isolated directly from whole blood by density gradient centrifugation.Fifteen SCID mice were randomly divided into two groups.Group A was control group,and in group B there were 12 mice intraperitoneally injected with 2×107 human PBL and subcutaneously injected with 5×106 ECA109 cells.The rate of tumor transplantation,tumor growth,metastasis and histological features were observed.After 3,4,5,6 weeks of engraftment,the level of IgG in mouse serum and the spleen weight were detected.Results The successful rate of tumor transplantation was 100 ％.Metastasis was not found.After 3,4,5,6 weeks of engraftment,the spleen weight in group B were (55.44±4.45) mg,(88.62±2.24) mg,[(125.98±2.19) mg] (P ＜ 0.05) and (213.71±2.96) mg,which had statistical significance compared with the control group (41.87±2.97) mg.The level of IgG was significantly higher than that in control group (P ＜ 0.01).Conclusion The experimental results demonstrate that human esophageal cancer models have been established in Hu-PBL-SCID mice.

BACKGROUND: Human β-defensin is mainly located in various tissues epidermis or epithelium, also exists in ocular surface, but its ocular surface and its role of ocular surface diseases remain poorly understood.OBJECTIVE: To observe the distribution of human β-defensins in ocular surface tissue, and to analyze their potential effects on ocular surface disease. DESIGN, TIME AND SETTING: In vitro controlled observation with regard to ocular surface tissue was performed at the Central Laboratory, Xinhua Hospital, Medical College of Shanghai Jiao Tong University and Cell Biochemistry Institute, Chinese Academy of Sciences Shanghai Branch, between October 2006 and December 2007.MATERIALS: A total of 18 inflammatory conjunctival specimens consisted of 6 pterygium surface bulbar conjunctiva, 4 bulbar conjunctival cysts, 4 acid burn conjunctiva, 2 thermal burn conjunctiva and 2 conjunctival granuloma; 15 inflammatory corneal specimens included 6 viral keratitis, 4 fungal keratitis, 3 bacterial keratitis and 2 eye removal following corneal perforation; 9 cadaver normal bulbar conjunctiva samples, 8 cadaver normal corneal samples. METHODS: RT-PCR method and immunohistochemistry were applied to detect human β-defensin expression in 50 samples. MAIN OUTCOME MEASURES: Distribution and location of human β-defensin proteins in normal and inflammatory ocular surface tissues. RESULTS: RT-PCR showed that human β-defensin 1 and human β-defensin 3 were positive in all of the tested samples, whereas human β-defensin 2 existed in a majority of inflammatory ocular surface tissues and no expression was observed in normal ocular surface tissues. Immunohistochemistry analysis revealed most of inflammatory ocular surface tissues expressed human β-defensins 1 and 2, distributing in epithelial cell layer and predominantly in basal lamina, occasionally infiltration of stromal cells was observed, only a small number of human β-defensin 2 expression was absent; normal cornea and conjunctiva samples presented with human β-defensin 1 expression, distributing in epithelial cells and predominantly in basal lamina, only few expressed human β-defensin 2.CONCLUSION: Human β-defensin 1 and 3 appear to be constitutively expressed in surface epithelial cells and basal lamina of normal and inflammatory ocular surface tissues, while human β-defensin 2 may be induced to express in the majority of inflammatory ocular surface tissues. Three human β-defensins expression plays a pivotal role in preventing ocular surface infection and promoting ocular surface injury repair.