Objective To observe pathological process of intestinal epithelial cells subjected to ischemia,ischemia/reperfusion injury and inflammation simulated hypoxia/reoxygenation (H/R) and lipopolysaccharide (LPS) challenged human fetal normal colonic cell (FHC) line in vivo,and to observe the changes when the assaulted intestinal epithelial cells were treated with emodin,in order to explore the possible intervention targets of emodin.Methods Normoxia group:the FHC cells were cultured in 95％ air and 5％ CO2 at 37 ℃.Hypoxia (H) group:the cells were cultured with a mixed anaerobic gas of 1％ O2,5％ CO2 and 94％ N2 at 37 ℃ for 1,2,3,4 hours.H + LPS group:the cells were cultured in hypoxic condition as H group with simultaneous challenge of LPS (1 mg/L).H/R group:the cells were cultured in hypoxia for 3 hours followed by reoxygenation for 1,2,3 and 4 hours,respectively.H/R + LPS group:the cells were cultured in H/R as H/R group and LPS (1 mg/L) simultaneously.Emodin intervention group:the cells were cultured in H3 h/R2 h + LPS and emodin (20,40,60,80 μmol/L) simultaneously.The variation trends of phosphorylation nuclear factor-κB profilin-o (pIκB-α),phosphorylation NF-κBp65 (pNF-κBp65) and their downstream target gene cyclooxygenase-2 (COX-2),and hypoxia-inducible factor-1α (HIF-1 α) were determined by Western Blot.The morphological changes in intestinal epithelium in different groups were observed using light microscope.The effect of emodin on the proliferation of intestinal epithelial cell was measured by methyl thiazolyl tetrazolium (MTT) assay.Results ① H group:the expressions of pIκB-α,pNF-κBp65 and COX-2 were upregulated,peaking at H1 h (0.350 ± 0.018,1.083 ± 0.054,0.903 ± 0.045),and then they gradually lowered (F value was 3.011,7.247,5.754,P value was 0.013,0.000,0.005,respectively).The expression of HIF-1 α peaked at H3 h (1.511 ± 0.076),but there was no significant difference among different groups (F=1.881,P=0.062).H + LPS group:the expressions of pIκB-α,pNF-κBp65,COX-2,HIF-1α were increased with elongation of duration of hypoxia,and a maximal induction was observed at H3 h (0.504 ± 0.025,1.255 ± 0.063,0.812 ± 0.041,1.209 ± 0.075,F value was 2.683,8.774,9.765,2.432,and P value was 0.011,0.000,0.000,0.026,respectively).H/R group:with the prolonged duration of reoxygenation,the expressions of NF-κB signaling pathway proteins (pIκB-α,pNF-κBp65,COX-2) were decreased and dropped to nadir at H3 h/R4 h (0.712 ± 0.034,1.202 ± 0.048,0.691 ± 0.042,F value was 1.923,6.765,2.719,and P value was 0.063,0.000,0.016,respectively).Compared with H group,HIF-1α was decreased with a prolonged duration of reoxygenation in H/R group,but there was no significant difference in value among different time points (F=1.280,P=0.081).H/R + LPS group:pIκB-o,pNF-κBp65,COX-2,HIF-1α showed no sign of degradation with the prolonged duration of reoxygenation,and their expression increased to maximum analogously at R2-3 h (3.302 ± 0.061,2.315 ± 0.055,2.017 ± 0.043,2.413 ± 0.098,Fvalue was 4.614,1.652,5.970,2.076,and Pvalue was 0.001,0.067,0.000,0.037,respectively).Emodin group:emodin when co-treated with H/R + LPS inhibited the expression of HIF-1o and NF-κB pathways with a dose-effect relationship (P＜0.05 or P＜0.01).Emodin at the dose of 80 μmol/L showed most marked inhibition (2.599 ± 0.130,1.772 ± 0.089,2.590 ± 0.129,2.518 ± 0.125).However,after treatment of emodin did not show such effect.② After treatment with H/R + LPS,there were morphological changes in cells:vacuoles,deformation and fusion.The speed of cell growth became much slower compared with H group.③ Emodin (20-80 μmol/L) had no significant effect on cell proliferation.Although emodin produced biological effect in this concentration range,it had no cellular toxicity.Conclusions Both hypoxia and inflammation can activate the hypoxia pathway of HIF-1α and the pro-inflammatory pathway of NF-κB,but different stimuli cause varying degrees of activation in these two pathways.In H/R group,both pathways were weakened during reoxygenation.However,in H/R + LPS group,the proteins remained to show a relatively high expression during the process of reoxygenation.This may be related to the pathophysiological mechanism of intestinal ischemia/reperfusion injury:hypoxia/reperfusion injury and LPS act together to destroy the intestinal epithelial cells and induce gut-derived sepsis.Emodin may inhibit inflammation by blocking HIF-1α/NF-κB-COX-2 signaling pathways.

Objective To investigate the protective effect of basic fibroblast growth factor(bFGF) gene modified mesenchymal stem cells(MSCs-bFGF)on cerebral isehemia in rats.Methods MSCs or MSCs-bFGF were transplanted into rat models of focal cerebral ischemia by intravenous injection.The neurological deficits and infarction volumes were evaluated,and the survival rate and differentiation of grafted MSCs were observed by double immunofiuoreseent labeling.Results In the rat cerebral ischemic model, both MSCs and MSCs-bFGF showed protective effect on the rats in comparison with control group.However, the protective effect was more significant in MSCs-bFGF group.Double immunofluorescent staining showed the number of BrdU-labeled and NeuN co-expression cells in MSCs-bFGF treated animals(127.40?7.43 and 11.20?3.09)were much more than in those of MSCs treated animals.While there was no significant difference between MSCs-bFGF and MSCs group in the number of GFAP co-expression cells.Conclusion MSCs transplantation has protective effect on cerebral ischemia in rats.Basic fibroblast growth factor gene modified MSCs is more effective than MSCs in neuroproteetion.

Background Curcumin can eliminate free radical arising in the system and superoxide anion and further inhibit lipid peroxidation.Researches proved that curcumin has resisting effect on oxidative damage of lens,but its effect on the formation of age-related cataract is under clear.Objective This study was to investigate the effect of curcumin on the cataract induced by sodium selenite.Methods Thirty healthy clean SD rats of 12-day-old were selected.All the rats were randomly divided into the normal control group,model group and curcumin group and ten rats for every group.The selenite cataract models were established in the rats of model group and curcumin group by the injection of sodium selenite subcutaneously.Meanwhile,0.005 ％ curcumin lavaged the rats in the curcumin group daily for two weeks.Lens of the rats were examined under the slit lamp at 4,7,10,14 days after injection in all groups,and the opacification of lens was graded based on the He' criteria.The lenses of rats were obtained at the end of experiment and then the level of superoxide dismutase (SOD),the activities of malondialdehyde (MDA) and glutathione peroxidase(GSH-Px) in rat lens were detected using biochemical assay method.The use of experimental animals followed the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results The lenses were transparent throughout the experimental duration in the blank control group.Compared with model group,the development of the Ⅲ,Ⅳ,Ⅴ grade of cataract was slow down in the curcumin group with the significant difference( P＜0.05 ).The content of MDA,SOD and GSH-Px activity in lens appeared considerable differences among 3 groups ( MDA:F =215.42,P＜0.01 ; SOD:F =46.83,P＜ 0.01 ; GSH-Px:F=44.29,P＜0.01 ).The level of SOD was significantly reduced in the model group and the curcumin group compered with blank control group ( P ＜ 0.05 ) and that in the curcumin group was higher than the model group ( P＜0.05 ).The MDA level in the model group and the curcumin group was obviously rised in comparison with blank control group( P＜0.05 ),but that in the curcumin group lowed in comparison to model group( P＜0.05 ).GSH-Px activity in the curcumin group was significantly lower than that in blank control group and higher than that in the model group( P ＜ 0.05 ).Conclusions The curcumin can effectively delay rather than the cataract formation induced by sodium selenite in SD rat by improved the resistance of oxidation in the lens.

OBJECTIVETo summarize clinical outcomes of locking compression plate (LCP) combined with minimally invasive percutaneous plate osteosynthesis (MIPPO) for the treatment of Pilon fracture.

METHODSFrom January 2009 to December 2012, Pilon fracture patients treated by LCP with MIPPO were retrospectively analyzed. All open fractures, pathologic fractures and those who had limb vascular disease or nerve injury were excluded. Thirty-eight patients were enrolled, including 29 males and 9 females aged from 21 to 78 years old with an average of 48 years old. According to AO classification, 20 cases were type B, 18 cases were type C. Operative time, blood loss, reduction quality, time of fracture healing complications and postoperative ankle joint function were applied for evaluating clinical outcomes, AOFAS scoring were used for assessing postoperative clinical effects.

RESULTSAll patients were followed up from 13 to 24 months (averaged 18 months). All patients obtained bone union without any plate failures or loss of fixation/reduction. One patient occurred superficial wound infection, and resolved with antibiotics and local wound care. Postoperative average AOFAS score was 81 (ranged 65 to 97).

CONCLUSIONLCP with MIPPO for Pilon fratcure has advantages of less invasion, fewer complications and satisfactory ankle function.

Adult ; Aged ; Bone Plates ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Retrospective Studies ; Tibial Fractures ; surgery

Generation of therapeutic angiogenesis to enhance vascularization in the ischemic tissues is a method for treating ischemic tissues in atherosclerotic cardiovascular artery disease. The chemokine stromal cell-derived factor-1 (SDF-1) and its receptor (CXC chemokine receptor 4, CXCR4) play a critical role in the process of post-natal neovascularization. The SDF-1-CXCR4 axis is a potential mechanism for the treatment of ischemic limb. Here, we investigated the role of CXCR4 in bone marrow cells (BMCs) in neovascularization induced by tumor cells and the supernatant of culture media in a murine hind-limb ischemia model which was made by resecting femoral artery and vein. After the injection of mouse melanoma cells B16-F10 (1x10(6) cells in 0.1 mL at the operation day, s.c.) into the abdomen or the cell culture supernatant (0.1 mL/d for 21 d after operation, i.m.) into the ischemic abductor muscle, the CXCR4 positive BMCs were analyzed by flow cytometry. The perfusion of the ischemic limb was evaluated by laser Doppler perfusion imaging (LDPI) on 7, 14 and 21 d after vascular injury operation. Capillary endothelial alkaline phosphatase (AP) was stained to quantify the presence of capillaries, and histological method was used to evaluate the capillary density as a measure of neovascularization in ischemic tissues. The proportions of CXCR4 positive BMCs were notably higher in ischemic limb injected with tumor cells or the supernatant compared to those in the control group (P<0.05). Injection of tumor cells or the supernatant resulted in significantly improved perfusion as measured by LDPI perfusion ratios on 7, 14 and 21 d after femoral artery and vein resection in mice, compared to the controls (P<0.05). Tissue samples harvested from the lower calf muscle at day 21 demonstrated increased capillary densities in mice receiving tumor cells (0.81+/-0.13) or the supernatant (0.63+/-0.05), compared with those in control group (0.44+/-0.09, P<0.05). In conclusion, the injection of B16-F10 tumor cells or the supernatant induces the increase of CXCR4 positive cells in BMCs and the improvement of in vivo neovasculogenesis in mouse ischemic limb.
Animals ; Bone Marrow Cells ; metabolism ; Capillaries ; Cell Culture Techniques ; Chemokine CXCL12 ; metabolism ; Culture Media, Conditioned ; pharmacology ; Disease Models, Animal ; Endothelium, Vascular ; Hindlimb ; blood supply ; Ischemia ; physiopathology ; Melanoma, Experimental ; Mice ; Muscle, Skeletal ; blood supply ; Neovascularization, Physiologic ; Receptors, CXCR4 ; metabolism

For microbial production of lycopene, the lycopene synthetic genes from Pantoea agglomerans were integrated into Saccharomyces cerevisiae strain BY4742, to obtain strain ZD-L-000 for production of 0.17 mg · L(-1) lycopene. Improving supplies of isoprenoid precursors was then investigated for increasing lycopene production. Four key genes were chosen to be overexpressed, inclu- ding truncated 3-hydroxy-3-methylglutaryl-CoA reductase gene (tHMG1), which is the major rate-limiting enzyme in the mevalonate (MVA) pathway, a mutated global regulatory factor gene (upc2.1), a fusion gene of FPP synthase (ERG20) and endogenous GGPP synthase (BTS1), which is a key enzyme in the diterpenoid synthetic pathway, and GGPP synthase gene (SaGGPS) from Sulfolobus acidocaldarius. Over-expression of upc2.1 could not improve lycopene production, while over-expression of tHMGI , BTS1-ERG20 and SaGGPS genes led to 2-, 16. 9- and20. 5-fold increase of lycopene production, respectively. In addition, three effective genes, tHMG1, BTS1-ERG20 and SaGGPS, were integrated into rDNA sites of ZD-L-000, resulting in strain ZD-L-201 for production of 13.23 mg · L(-1) lycopene, which was 77-fold higher than that of the parent strain. Finally, two-phase extractive fermentation was performed. The titer of lycopene increased 10-fold to 135.21 mg · L(-1). The engineered yeast strains obtained in this work provided the basis for fermentative production of lycopene.
Bacterial Proteins ; genetics ; metabolism ; Biosynthetic Pathways ; Carotenoids ; biosynthesis ; Genes, Synthetic ; Genetic Engineering ; Pantoea ; enzymology ; genetics ; Saccharomyces cerevisiae ; genetics ; metabolism

AIM: The present study was undertaken to explore the effects of ?-MSH on partial biological activities of LPS. METHODS:Colorimetric method was used for the measurement of hydrogen peroxide(H_2O_2) production in mouse peritoneal macrophages, the apoptosis of polymorphonuclear leukocytes(PMNs) and the binding of LPS to monocytes were studied with flow cytometry. RESULTS: It was found that LPS strongly stimulated macrophages to release H_2O_2. When macrophages were cultured with ?-MSH in the presence of LPS, the H_2O_2 release was markedly suppressed (P0.05). In the presence of LPS, however, ?-MSH significantly promoted the apoptosis of PMNs (P

OBJECTIVETo observe the effect of octyl-a-cyanoacrylate upon bone healing and its degradation in vitro after middle tibial transverse fracture in rabbitsì and to establish treatment of higher efficacy with the application of octyl-a-cyanoacrylate.

METHODSMiddle tibial transverse fracture model of New Zealand rabbits was established. In the experimental group, internal fixation with 2 mm Kirschner wires was performed and the broken ends were fixed with octyl-a-cyanoacrylate. In the control group, only internal fixation with 2 mm Kirschner wires was conducted. Animals were killed at preset time intervals of 2, 4, 6, 8, 10 and 12 weeks postoperatively and samples were harvested.

RESULTSTwo weeks after operation, clear fracture lines were observed in both the experimental and the control groups. Fibrous soft tissue connection was noted between the broken ends and there was soft tissue adhesion around the fracture site. There was no callus formation and the broken ends were surrounded by adhesive soft tissues. Obvious external callus formation was confirmed at 8 weeks after operation in both groups with partial disappearance of fracture lines. Ten and twelve weeks after the operation, fracture lines disappeared completely and there was obvious external callus formation and bone union. In the fourth week, fibrous cells and chondrocytes were found to grow into the colloid and surround it at the 6th week. The adhesive material was degraded and gradually absorbed at the 8th week. Chondrification was observed.

CONCLUSIONSTwo weeks after fixation for tibial fracture in rabbits, octyl-a-cyanoacrylate begins in vivo degradation. Chondrocytes and fibrocytes gradually grow into the degradation area and surround the adhesive material, which broke into pieces at 8 weeks. Complete degradation and disappearance of the adhesive material is present between 10 and 12 weeks. No barrier effect hampering fracture healing is noted.

Adhesives ; therapeutic use ; Animals ; Cyanoacrylates ; therapeutic use ; Rabbits ; Radiography ; Tibial Fractures ; diagnostic imaging ; pathology ; therapy

Objective:To explore the barriers of nutritional care for spouses of elderly patients with stroke, and provide reference for the home nutrition education management program and related interventions to improve the nutritional care quality of elderly spouses.Methods:Semi-structured interviews were conducted for spouses of 13 elderly patients with stroke. The Colaizzi 7-step analysis was used for analysis.Results:The barriers could be summarized into three generic categories and seven themes. Cognitive factors included poor nutrition cognitive awareness, and weakened self-maintenance. Behavioral factors included poor compliance, improper diet adjustment, and invalid personal response. Environmental factors included delayed social support and limited access to information.Conclusions:Paying attention to the barrier factors of nutritional care for spouses of elderly patients with stroke can help us understand the difficulties and problems of spouses, and provide reference for the effective implementation of family nutrition education and management.

OBJECTIVETo study the diagnostic value of heart rate variability (HRV) in heart dysfunction in children with beta-thalassemia major (β-TM)by examining the changes of HRV in β-TM children.

METHODSA 24 hours Holter monitoring electrocardiogram (Holter) was performed in 21 children with β-TM and 15 healthy children (control group). The time domain and frequency domain indexes of HRV in the two groups were compared. The correlation between serum ferritin levels and HRV was evaluated.

RESULTSThe time domain indexes SDNN, rMSSD and PNN50 and the frequency domain indexes very low frequency (VLF), low frequency (LF) and high frequency (HF) in the β-TM group were significantly lower than in the control group (P<0.05). There was no correlation between serum ferritin level and HRV in children with β-TM.

CONCLUSIONSThe autonomic nerve dysfunction exists in children with β-TM. HRV analysis is useful in the prediction of early cardiac dysfunction in children with β-TM.

Adolescent ; Child ; Female ; Ferritins ; blood ; Heart Rate ; physiology ; Humans ; Male ; beta-Thalassemia ; blood ; physiopathology