This study examined the effect of sulodexide on podocyte injury in rats with adriamycin nephropathy (AN). A total of 36 healthy male SD rats were randomly assigned to three groups: control group, AN group and sulodexide treatment group. Rat models of AN were established by a single tail intravenous injection of adriamycin (6.5 mg/kg) in both AN group and sulodexide treatment group. Sulodexide (10 mg/kg) was administered the rats in the treatment group once daily by garage from the first day of model establishment until the 14th day or the 28th day. Samples of 24-h urine and renal cortex tissues were harvested at day 14, 28 after the model establishment. Excretion of 24-h urinary protein was measured by Coomassie brilliant blue method. The pathological changes in renal tissues were observed by light microscopy and electron microscopy respectively. Heparanase mRNA was detected by RT-PCR. Expressions of desmin, CD2AP and heparanase were determined by immunohistological staining. The results showed that the expressions of heparanase mRNA and protein were increased in the glomeruli of AN rats at day 14 and 28 after the model establishment, which was accompanied by the increased expression of desmin and CD2AP. The mRNA and protein expression of heparanase was decreased in the sulodexide-treated rats as compared with AN rats at day 14 and 28. And, the protein expression of desmin and CD2AP was reduced as with heparanase in the sulodexide- treated rats. Proteinuria and podocyte foot process effacement were alleviated in the AN rats after sulodexide treatment. There was a positive correlation between the expression of heparanase and the expression of desmin and CD2AP (as well as 24-h urinary protein excretion). It was concluded that increased heparanase is involved in podocyte injury. Sulodexide can maintain and restore podocyte morphology by inhibiting the expression of heparanase in AN.

Objective To access the effects of aldosterone (ALD) and its receptor antagonist- spironolactone (SPI) on the production of reactive oxygen species (ROS) and apoptosis in podocytes and to explore the possible mechanism involved. Methods Conditionally immortalized mouse podocytes were divided into control group, ALD group, SPI group and SPI +ALD group. The level of ROS production and the expression of nephrin protein were assayed by fluorescent spectrophotometry and indirect immunofluorescence technology. The apoptosis rate of podocytes was monitored by flow cytometry. The expression of bax and bcl-2 mRNA and protein was detected by RT-PCR and Western blotting methods. The anti-oxidant N-acetylcysteine (NAC)was applied to determine whether the effects of ALD on podocytes were mediated by ROS pathway.Results Compared with the control group, ALD significantly increased ROS production in podocytes (P＜0.05). SPI completely abolished the above-mentioned effect of ALD (P＜0.05). ALD induced the down-regulation of the expression of nephrin and the up-regulation of podocytes apoptosis (P＜0.05), which was accompanied with the elevated expression of bax mRNA and protein and the reduced expression of bcl-2 mRNA and protein (P＜0.05). SPI or NAC prevented the above-mentioned changes induced by ALD (P＜0.05). Conclusion ALD increases theexpression of pro-apoptotic factor (bax) but decreases the expression of anti-apoptotic factor (bcl-2)to induce podocytes apoptosis through the mineralocorticoid receptor (MR) possibly via the mechanisms involving ROS pathway.

Objective To investigate the feasibility of combined laparoscopic and thoracoscopic esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity.Methods We retrospectively analyzed the clinical data of 38 patients who underwent esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity from October 2011 to August 2012.To remove the stomach in laparoscopic and the esophagus in thoracoscopy.The main portion of a gastric conduit is created using three to four firings of a linear stapler(Ethicon Endo-surgery,Cincinati,OH) and jejunum stoma.Gastric conduit was pulled into the chest cavity and anastomosed to the esophagus.Results The average operative time was 280 minutes,the mean operative blood loss was 120 ml.No patient required laparotomy.No pulmonary complications or anastomotic leaks occurred.One had gastric retention,another one had chylous hydrothorax.All patients were cured,no one dead in hospital.Conclusion Combined laparoscopic and thoracoscopic esophagectomy for esophageal carcinoma and gastro-esophageal anastomosis in right thoracic cavity is technically feasible and safe,minimized trauma,less operative blood loss and quick recovery.

Objective To study the indication,feasibility and short-term efficacy of combined thoracoscopic and laparoscopic radical esophagectomy for the treatment of esophageal cancer.Methods Retrospective medical records analysis was conducted for 139 esophageal cancer patients who underwent combined thoracoscopic and laparoscopic esophagectomy in our department from December 2009 to August 2011.The tumors were located in upper esophagus in 16 cases,middle esophagus in 107 cases,and lower esophagus in 16 cases.The surgery started with the thoracoscopic mobilization of thoracic esophagus and lymph nodes dissection,which were followed by the laparoscopic stomach mobilization and gastroesophageal anastomosis in left neck.Postoperative pathological staging identified stage Ⅰ esophageal cancer in 25 cases ( stage Ⅰ a:13 cases,stage Ⅰ b:12 cases),stage Ⅱ esophageal cancer in 71 cases,stage Ⅲ esophageal cancer in 31 cases ( stage Ⅲ a:16 cases,stage Ⅲ b:15 cases) and stage Ⅳ esophageal cancer in 12 cases.Results Except for open conversions in 4 cases (2.9％),all surgical operations were completed smoothly.Postoperative anastomotic leak was found in 6 cases(4.3％ ),chylothorax in 1 case(0.7％ ),arrhythmia in 4 cases(2.9％ ),and dumping syndrome in 1 case( 0.7％ ).All of these complicated cases fully recovered after conservative treatments.Postoperative lung infection was found 11 cases (7.9％),3 of whom required tracheotomy and assisted ventilation and 1 case died as a result of the infection (mortality rate:0.7％ ).Ten cases(7.2％ ) presented with hoarseness postoperatively.Out of the 139 cases,130 cases were successfully followed up with durations ranged from 1 to 20 months,during of time the esophageal cancer spread to liver in 2 cases,celiac lymph nodes in 4 cases,lung in 2 cases,and bone in 1 case.Ten cases died,and all remaining cases remained alive during the follow up.The one-year survival rate was 88.9％ for these cases.Conclusion Combined thoracoscopic and laparoscopic radical esophagectomy is a technically safe and feasible treatment for esophageal cancer.The short-term efficacy results are satisfactory.This technique is indicated not only for early and middle stage esophageal cancer,but also for some of the advanced esophageal cancer cases.

OBJECTIVETo study the transport mechanism of honokiol in Caco-2 cell model.

METHODThe analysis was performed on a Kromasil 100-5 C18 column (4.5 mm x 250 mm, 5 microm) eluted with acetonitrile-water (70: 30) as mobile phase. The detection wavelength was set at 203 nm. Two-way transport of honokiol was studied by using Caco-2 cell model, and the effects of time, drug concentration, inhibitor, pH, temperature on the transport of honokiol was investigated. The drug concentrations were determined by high performance liquid chromatography(HPLC) and used to calculate the apparent permeability coefficient.

RESULTThe standard curve of honokiol was Y = 24 044X - 3 763.6 (r = 0.999 8), and the detection limit was 0.04 micromol x L(-1). In Caco-2 cell model, the transport amounts from the top side to the base side of were more than that from the base side to the top side under the same concentration. The transport amounts increased with time both in AP --> BL and BL --> AP directions. Verapamil could improve the transport amounts of AP --> BL. There were no effects of pH on the transport of AP --> BL. Both in AP --> BL and BL --> AP directions, the transport showed temperature dependence.

CONCLUSIONHonokiol is transported through the intestinal mucosa via a passive diffusion mechanism primarily, coexisting with a carrier-mediated transport, at the same time effected by P-gp.

Biological Transport ; Biphenyl Compounds ; analysis ; pharmacokinetics ; Caco-2 Cells ; Chromatography, High Pressure Liquid ; methods ; Diffusion ; Humans ; Lignans ; analysis ; pharmacokinetics ; Permeability

This study examined the effect of sulodexide on podocyte injury in rats with adriamycin nephropathy (AN). A total of 36 healthy male SD rats were randomly assigned to three groups:control group,AN group and sulodexide treatment group. Rat models of AN were established by a single tail intravenous injection of adriamycin (6.5 mg/kg) in both AN group and sulodexide treatment group. Sulodexide (10 mg/kg) was administered the rats in the treatment group once daily by garage from the fast day of model establishment until the 14th day or the 28th day. Samples of 24-h urine and renal cortex tissues were harvested at day 14,28 after the model establishment. Excretion of 24-h urinary protein was measured by Coomassie brilliant blue method. The pathological changes in renal tissues were observed by light microscopy and electron microscopy respectively. Heparanase mRNA was detected by RT-PCR. Expressions of desmin,CD2AP and heparanase were determined by immunohistological staining. The results showed that the expressions of heparanase mRNA and proteinwere increased in the glomeruli of AN rats at day 14 and 28 after the model establishment,which was accompanied by the increased expression of desmin and CD2AP. The mRNA and protein expression of heparanase was decreased in the sulodexide-treated rats as compared with AN rats at day 14 and 28. And,the protein expression of desmin and CD2AP was reduced as with heparanase in the sulodexide-treated rats. Proteinuria and podocyte foot process effacement were alleviated in the AN rats after sulodexide treatment. There was a positive correlation between the expression of heparanase and the expression of desmin and CD2AP (as well as 24-h urinary protein excretion). It was concluded that increased heparanase is involved in podocyte injury. Sulodexide can maintain and restore podocyte morphology by inhibiting the expression of heparanase in AN.

OBJECTIVE:To prepare and characterize Fluorescent dye 1,1′-octacosyl-3,3,3′,3′-tetramethylindocarbocyanine iodide(DiR)-loading polyethylene glycol-poly lactic-co-glycolic acid(DiR-PEG-PLGA)nanocapsules,and to evaluate its biocompatibility in vitro. METHODS:Using PLGA and PEG-PLGA as carrier,DiR-PEG-PLGA nanocapsules were prepared by modified ultrasonic emulsification method. The particle size,Zeta potential,morphology,stability and fluorescence in vitro of nanocapsules were detected respectively. MTT assay was used to evaluate cytotoxicity in vitro of nanocapsules to human-derived HL7702 hepatocytes,and hemolysis test was carried out to investigate its hemolysis effects. RESULTS:Prepared DiR-PEG-PLGA nanocapsules were spherical with a clear core-shell structure. The average particle size was(507.53 ± 7.87)nm,polydispersity coetficient of particle size was 0.306 1±0.001 5 and Zeta potential was(-35.20±0.92)mV with good stability within 6 months under 4℃. Fluorescence signal intensity(y)of nanocapsules was increased linearly with DiR mass concentration(x)in vitro. The linear eguation was y=0.345 2x+0.433 4(R2=0.997 3).The toxicity of nanocapsules to HL7702 cells was between 0-1 degree,and no hemolytic effect was observed. CONCLUSIONS:The study successfully prepare fluorescent DiR-PEG-PLGA nanocapsules with high biocompatibility in vitro,which is further expected to become a safe optical drug carrier.

Possible mechanisms by which Polygonati rhizoma opposes atherosclerosis (AS) were identified by network pharmacology and molecular docking analyses. The Traditional Chinese Medicine Database (TCMD) and the Traditional Chinese Medicine Systems Pharmacology Database (TCMSP) were utilized to identify the likely active components of Polygonati rhizoma. The potential targets set of Polygonati rhizoma were predicted with the PharmaDB database and the Swiss TargetPrediction database. The targets set for AS was retrieved by OMIM, DisGeNET and NCBI Gene database. We used the STRING platform to construct a protein-protein interaction network of the intersectional targets and performed visual analysis in Cytoscape. The key targets of Polygonati rhizoma in AS were searched by network topology and the resulting GO and KEGG enrichment was analyzed by Clue GO. In addition, the key targets were verified by molecular docking in Discovery Studio 4.0. A total of 45 active ingredients and 51 potential targets were obtained in the treatment of AS. The results of the topology analysis included five key targets: serum albumin, mitogen-activated protein kinase 3, mitogen-activated protein kinase 1, proto-oncogene tyrosine-protein kinase Src and matrix metalloproteinase-9. The 131 GO items showed that the biological process mainly involved the steroid receptor, cell response to steroid stimulation, the phosphatidylinositol-3 kinase signal pathway, and others. The KEGG pathway analysis included 37 pathways, which were closely related to peroxisome proliferation activated receptor signaling pathway, platelet activation pathway, vascular endothelial growth factor pathway, hypoxia inducible factor pathway and adhesion connection pathway. The results of molecular docking proved that the combined activity of the components with potential key targets is excellent. This study proposes mechanisms by which Polygonati rhizoma might act to reverse or minimize AS and provides a scientific basis for clinical research on Polygonati rhizoma.