Objective To observe the expression of DR5 in the ocular tissues of uveitis rats induced by endotoxin,and study the relationship between the apoptosis of inflammatory cells and expression of TRAIL / DR5.Methods SD rats were randomly divided into 3 groups:Blank control group,normal saline injection group and endotoxin injection group.The endotoxin injection group was injected with lipopolysaccharide into the rat posterior foot pad to make endotoxin-induced uveitis animal model.There were no operations in the blank control group,and the subgroups were divided into 6 hours,12 hours,24 hours and 48 hours groups according to the time of injection.The ultrastructural changes of inflammatory cells and endothelial cells in iris capillaries were observed by transmission electron microscopy (TEM).The expression of DtR5 protein on inflammatory cells at different time after endotoxin induction was detected by SABC method.Results TEM showed that the microvilli of the capillary endothelial cells in the iris tissue of the blank control group and saline injection group had more obvious vesicles with no obvious abnormal structure and shape.The number of swallowed vesicles in the capillary endothelial cells injected with endotoxin was decreased at 6 hours group,and the number of vesicles in the infiltrating neutrophils and lymphocytes decreased.Neutrophils and lymphocytes appear chromatin condensation,vacuolar changes in the expression of apoptosis.Immunohistochemistry showed that the DR5 protein was negative in the iridocular epithelium of the blank control group and saline injection group.In the endotoxin injection group,the DR5 protein was weakly colored in the iris pigment epithelium and appeared on the inflammatory cells.The number of staining and the intensity of coloring in the 24 hours group were significantly higher than those in the 6 hours group,and the color density was 0.085 9 ± 0.019 6,there were statistical differences compared with 6 hours group,12 hours group and 48 hours group (all P ＜ 0.05).Conclusion TRAIL and its receptor DR5 may be involved in the apoptosis of inflammatory cells in endotoxin-induced uveitis.

Objective To investigate the hemodynamics and structural effect of rat endothelial progenitor cells (EPC) transplant on pulmonary artery hypertension (PAH) induced by monocrotaline(MCT) in rats.Methods EPCs were identified and marked.Twenty-one days after injection of EPCs,the pulmonary hemodynamic parame- ters,average pulmonary artery pressure (mPAP),right heart index were determined.The vascular endothelial cells and pulmonary vascular structural changes were verified by fluoresccuse microscope.Results Compared with the model,EPCs treatment(n=10) decreased mPAP significantly (mPAP,EPCs:25.9?0.7 mmHg vs model group:29.3?2.2 mmHg,P

Objective To investigate the effect of telmisartan on the oxidative stress induced by high glucose in human umbilical vein endothelial cell (HUVEC) in vitro.Methods HUVEC were cocultured with telmisartan (1?10~(-6) mol/L) and various concentration of glucose(5,30 mmol/L) for 0,12,24,36,48 h respectively. The level of MDA in the supernatants of cultured endothelial cells was measured by thiobarbituric acid test,SOD was determined by xanthine oxidase test.The protein expression of peroxisome proliferator activated receptors ? (PPAR-?) in HUVEC 24 h was assessed by Western blot after treatments.Results High glucose significantly increase the levels of MDA (before:1.2?0.06 vs after:1.6?0.1 mmol/mL,P

Background Doxycycline is a broad spectrum antibiotic,and it is frequently used in the treatment of ocular surface diseases.Objective The purpose of the present study was to investigate the effect of doxycycline on the inhibition of cell proliferation in bovine corneal myofibroblasts in vitro and assess its contribution to ocular surface repairing mechanism.Methods Six fresh bovine corneas were collected.The corneal stromal layer was isolated by two-step method of 1.0 g/L and 2.0 g/L collegenase-1.Isolated cells were plated at mantaryay culture flask in 10％ FBS of RPMI-1640.Vimentin and alpha-smooth muscle actin (α-SMA) organization were evaluated by immunocytochemistry,and the cells with influoresccence staining for vimentin and α-SMA were identified as the corneal myofibroblasts.Doxycycline at the concentrations of 10,20,40,60,80 mg/L was added to the medium,respectively,in different concentrations of doxycycline groups.Dexamethasone (120 mg/L)was used in the same way in the positive control group,and no drug was used in the negative control group.Cell proliferation was evaluated by MTT and the cell cycle was analyzed by BD FACScan flow cytometer assay 24 hours and 48 hours after addition of any drug.Results The cells grew well and showed the positive response for vimentin and α-SMA.MTT assay showed that the A570values of bovine corneal myofibroblasts were gradually declined with the increase of the concentration of doxycycline and lapse of active time,showing statistically significant difference (Fconcentration =1233.778,P＜0.001 ; Ftime =227.564,P ＜ 0.001).And the difference between the two factors was also statistically significant (Ftime*concentration =51.656,P＜0.001).Flow cytometry cell cycle analysis showed that 24 hours after 10,20,40,60,80 mg/L doxycycline treated,the perentage of of corneal myofibroblast cell in G0-G1 phase was 82.85％,84.36％,85.18％,87.12 ％ and 89.31％,showing significant increase in comparison with 63.89％ of the negative control group (all P＜0.05),and that of 40 mg/L doxycycline group was near the positive control group.Forty-eight hours after 10,20,40,60,80 mg/L doxycycline treated,the perentage of of corneal myofibroblast cell in G0-G1 phase was 82.78％,86.15％,88.23％,89.57％,93.00％,with significant increase in comparison with 70.17％ of the negative control group (all P ＜ 0.01),and that of 40 mg/L doxycycline group was near the positive control group.Conclusions The growth of the bovine corneal myofibroblasts is inhibited by doxycycline in time-and dosedependent manner in the range from 10 mg/L to 80 mg/L,and 40 mg/L of doxycycline has an obviously inhibitory action as 120 mg/L dexamethasone.

A quantitative HPLC-DAD method was developed for simultaneous determination of N-trans-p-coumaroyloctopamine and N-trans-p-coumaroyltyramine in Solani Melongenae Radix from different cultivation regions in China The separation was performed on an Agilent Eclipse XDB C18 column (4.6 mm x 250 mm, 5 microm) at 30 degrees C with a gradient elution of methanol and 0.1% formic acid in water as mobile phase. The flow rate was set at 1.0 mL x min(-1) and the detection wavelength was 300 nm. The calibration curves of N-trans-p-coumaroyloctopamine and N-trans-p-coumaroyltyramine were linear over the ranges of 2.84-68.16, 3.10-74.40 mg x L(-1), and the average recoveries (n = 9) were 99.30% and 102.8%, respectively. The developed method was successfully applied for the analysis of sixteen samples from different cultivation regions in China, which indicated that the method is simple, rapid, accurate, and reliable for quality evaluation of Solani Melongenae Radix.
Amides ; analysis ; China ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Plant Roots ; chemistry ; Solanaceae ; chemistry ; classification

Objectives To detect the expression of serum high mobility group box-1 (HMGB1) and explore its changes in rats with acute necrotizing pancreatitis (ANP).Methods Intraperitoneal injection of 20％ L-arginine in the dosage of 250 mg/100 g twice every 1 hour was used to establish ANP rat model.Intraperitoneal injection of normal saline solution in equal volume was performed in control rats.Rats were sacrificed at 6 h,18 h,24 h,36 h,48 h,72 h and 96 h after injection.Blood samples were collected to detect serum amylase and HMGB1 level.Pancreatic tissue was collected for pathological examination.Realtime PCR was applied to detect the mRNA expression of HMGB1 in pancreatic tissue.Werstem blot was used to determine HMGB1 protein expression in pancreatic tissue.Results Serum amylase level began to increase at 6 h after modeling,reached the peak at 18 h [(5 070 ± 603) U/L] and returned to normal level after 48 h.Serum amylase activity at 6 h and 18 h in ANP group was much higher than that in control group (1 844 ± 181)U/L(P＜0.05).The expression of HMGB1 began to increase at 6 h,reached to the peak at 36 h [(288.5 ±42.1)μg/L],and then decreased gradually.HMGB1 expressions at each time point in ANP group were significantly higher than those in control group (31.6 ± 10.1) μg/L],and the differences were statistically significant (all P ＜ 0.05).Pathological scores in pancreatic tissues in ANP group were higher than those in control group 0.38 ± 0.52,and the differences were statistically significant (P ＜ 0.05).HMGB1 mRNA expressions at t 6 h,18 h,24 h,36 h,48 h,72 h and 96 h in ANP group were 1.23 ±0.25,2.60 ± 0.46,3.23 ± 0.34,4.77 ± 0.66,2.88 ± 0.56,2.05 ± 0.20,1.33 ± 0.28,which were significantly higher than those in control group 0.44 ± 0.09,and the relative expression of HMGB1 in ANP group at 36 h was significantly higher than those at other time points (all P ＜ 0.05).HMGB1 protein expression in pancreatic tissue in ANP group at 6 h,18 h,36 h,72 h were 1.14 ±0.02,1.15 ±0.01,1.22 ±0.01,1.22 ±0.04,which obviously higher than those in control group(1.0),and HMGB1 expression in ANP group at 36 h was higher than those at other time points (all P ＜ 0.05).Conclusions HMGB1 may participate in systematic inflammation as one of the late inflammatory mediators during ANP.

Objective To explore the value of monitoring in vertebral and spinal cord operation by the real-time monitoring of the spinal cord in the process of operation.Methods Nineteen cases were included by the real-time monitoring of spinal cord in the process of operation.Japanese Orthopaedic Association(JOA) score was used to evaluate the clinical efficacy during following-up before and discharge after operation,6 months and 12 months after operation.Results Intraoperative ultrasound monitoring in the 19 patients had no symptom of spinal cord injury and other complications within the plant without loosening fracture occurred were bony fusion; the average JOA scores in the patients before operation,discharge after operation,6 months and 12 months after operation were 8.80±1.60,14.00±1.57,14.60±1.61 and 14.80±1.58,respectively.The improvement rate of JOA score for discharge after operation,6 months and 12 months after operation were individually 51.6%,61.3%,and 64.5%.The JOA score after operation was obviously higher than that before operation.The significant difference between the two groups was observed in the experiment (P<0.05).The cervical spinal canal sagittal diameter significantly increased after operation (mean sagittal diameter of the expansion of 6.6 mm),compared with that before the trial (P<0.05).Conclusion The real-time monitoring with ultrasonography during operation can identify the change of spinal cord and provide the image evidence of operative efficiency to avoid the injury in vertebral and spinal cord operation.

Objective To study the effects of total extracts of Averrhoa carambola L.( Oxalidaceae ) roots ( TEACLR ) in hyperlipidemia rats. Methods Sixty SD rats ( male and female in half ) were subjected to induction of hyperlipidemia by high -fat diet for established hyperlipi-demia rat model and were divided into the following groups:blank group, model group, positive control group ( simvastatin 2.5 mg? kg-1? d -1 orally) , TEACLR high, middle and low doses groups.Hyperlipidemia rats were treated with TEACLR, at doses of 1200, 600 and 300 mg? kg-1? d-1 for 5 weeks by gavages, respectively.After 5-week of treatment with drugs, blood sample and liver were collected for determi-nation.Results In contrast to the model group, TEACLR could reduce the contents of cholesterol, triglyceride, low-density lipoprotein, alka-line phosphatase in rat serum and liver malondialdehyde reduced obvious-ly [ ( 2.48 ±0.43 ) , ( 0.86 ±0.44 ) , ( 0.86 ±0.43 ) mmol? L-1 and (165.55 ±71.71 ) U? L-1 , ( 1.38 ±0.42 ) nmol? prot-1 , respective-ly] .The level of high -density lipoprotein and glutathione elevated evidently [(1.67 ±0.32)mmol? L-1,(3.43 ±0.78)nmol? prot -1]. Liver coefficient was significantly lowered ( 2.92 ±0.03 ) .TEACLR middle dose group could reduce the contents of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) obviously[(38.91 ±10.47),(99.91 ±30.74) U? L-1 ].TEACLR high dose group could increase the contents of superoxide dismutase evidently(24.44 ±3.13) nmol? prot -1 .Comparison with blank group and positive control group, there was no significant difference in TEACLR middle dose group.Conclusion TEACLR is proven to be able to decrease the level of hyperlipidemia markedly, which might be used as an effective therapeutic agent for hyperlipidemia and fatty liver.

BACKGROUND/AIMS: The aim of this study was to investigate whether genetic variations at positions -1082, -819, and -592 in the interleukin (IL)-10 promoter affect IL-10 production in children with irritable bowel syndrome (IBS). METHODS: Ninety-four children with IBS and 102 children as healthy controls (HCs) were enrolled. Genomic DNA was extracted, and IL-10 -1082, -819, and -592 polymorphisms were detected by direct sequencing from all participants. Peripheral blood mononuclear cells (PBMCs) from 46 IBS children and 38 HCs were isolated and cultured with and without 5 ng/mL Escherichia coli lipopolysaccharide (LPS). IL-10 levels in the culture supernatants were measured by enzyme-linked immunosorbent assay. RESULTS: There were no significant differences in the distribution of IL-10 -1082, -819, and -592 polymorphisms or in the allele and haplotype frequencies between IBS children and HCs. PBMCs from children with IBS had significantly lower IL-10 levels after LPS stimulation than PBMCs from HCs (p=0.011); however, LPS-induced IL-10 levels in PBMCs with different genotypes of -819 and -592 polymorphisms were not significantly different between IBS patients and HCs. CONCLUSIONS: Although significantly lower LPS-induced IL-10 production by PBMCs was noted, it is unlikely that IL-10 production was fully genetically determined in our IBS children. ClinicalTrials.gov identifier: NCT01131442.
Alleles ; Child ; DNA ; Escherichia coli ; Genetic Variation ; Genotype ; Haplotypes ; Humans ; Interleukin-10 ; Interleukins ; Irritable Bowel Syndrome

OBJECTIVETo develop a method for tumor segmentation on multi-modality magnetic resonance (MR) images based on parameter optimization of SVM model.

METHODSEach one of the 4 sub-classifiers was trained using the feature information in mono-modality MR images and applied to the corresponding modality images. The classification results differed due to different information in the selected support vectors of the mono-modality images. By modifying the weight values of the error data points, we chose the best weight values of the sub-classifier to obtain a weighed combination SVM classifier of multi-modalities for use in MR image segmentation.

RESULTSThis tumor image segmentation method was validated on the MR images of brain tumors in 34 patients and resulted in an average classification accuracy of 90.59%. Compared with the 4 mono-modality classifiers, multi-modality RBF kernel SVM classifiers increased the overall accuracy by 5.76%-20.11%.

CONCLUSIONThe proposed method combines multi-modality images with SVM classifiers to allow accurate tumor image segmentation from MR images with a high precision.