Objective To investigate the effect of silk fibroin scaffolds seeded with adipose mesenchymal stem cells (ADMSCs) in remodeling fiber for tunica albuginea defect in rabbits.Methods Fifty-six New Zealand rabbits were divided into 4 groups according to the random number table after a defect was created in the tunica albuginea:Group A (the defect was repaired with silk fibroin),Group B (with silk fibroin seeded with ADMSCs),Group C (with autologous tunica vaginalis) and Group D (left unrepaired),with 14 rabbits per group.Tunica albuginea sections were obtained for HE staining,Sirius red staining,Hart staining and immunofluorescence staining of macrophages at 6 and 12 weeks after surgery.Results At 12 weeks after surgery,HE staining revealed chaotically distributed new fiber ingrowth in Group D and orderly ingrowth in Groups A,B,and C.At 12 weeks after surgery,Sirius red staining revealed mean area of type Ⅰ collagen fibers was greater than that of type Ⅲ in Groups A (98 780 ±4 190 vs 51 177 ±5 464),B(94 855 ±9 010 vs 50 815 ±3 895),and C(99 860 ±6 015 vs 50 948 ± 6 595),but the difference in area of collagen fiber of the same type was insignificant among the three Groups.Moreover,less type Ⅰ collagen fibers (79 386 ±2 237) and more type Ⅲ collagen fibers (85 278 ± 2 645) were observed in Group D compared with other three Groups (P ＜ 0.01).At 12 weeks after surgery,Hart staining showed the mean area of elastic fibers in Groups A,B,C,and D was 2 805 ± 90,2 849 ±84,3 068 ±485,and 1 961 ±96 respectively.There was no statistical difference between Groups B and C,but less amount of fibers was observed in Group A (P ＜ 0.01) and least amount was observed in Group D (P ＜0.01).At 6 weeks after surgery,the number of infiltrating macrophages in Groups A,B,C and D was 4.10 ± 0.87,3.80 ± 0.78,3.70 ± 0.94,and 6.80 ± 1.63 respectively.There was no statistical difference in the number of macrophage infiltration among Groups A,B and C,but all were lower than that in Group D (P ＜ 0.01).Conclusion Silk fibroin seeded with ADMSCs is comparable to autologous grafts for repair of tunica albuginea defect in rabbits.

OBJECTIVEThe aim of the present study is to explore the effects of exhaustive exercise-induced oxidative stress on the antioxidant capacity and diformability of rat red blood cells.

METHODSRats were divided into three group (n = 10): sedentary control (C), exhaustive running exercise (ERE) and moderate running exercise (MRE) groups. Animals in the ERE group started treadmill running at a speed of 20 m/min speed with a 5% gradient, and reached a speed of 25 m/min with gradient 15% in 20 min. Running was continued until exhaustion. MRE group rats running at a speed of 20 m/min with a 5% gradient for 40 min. The levels of free thiol in erythrocyte membrane protein, lipidperoxidation levels and membrane protein components were analyzed. The red blood cell deformability of different groups was also observed.

RESULTSThe results showed that red blood cells were damaged by severe oxidative stress and the anti-oxidative capacity decreased significantly under exhaustive exercise conditions. Besides, lipid peroxidation and protein sulfhydryl cross-link based clustering of membrane were found after exhaustive exercise, and polymers high molecular weight (HMW) was formed. The elongation index (EI) was found to decline significantly in the ERE group compared with the C and MRE groups under shear stress (control group, 0.41 +/- 0.01 at 3 Pa and 0.571 +/- 0.008 at 30 Pa; ERE group, 0.314 +/- 0.013 at 3 Pa and 0.534 +/- 0.009 at 30 Pa; P < 0.05 and P < 0.01, respectively).

CONCLUSIONThese exercise-induced oxidative injure result in a significant decrease in deformability of rat erythrocytes, which in turn leads to dysfunction in the microcirculatory.

Animals ; Disease Models, Animal ; Erythrocyte Deformability ; Fatigue ; metabolism ; physiopathology ; Male ; Oxidative Stress ; Physical Conditioning, Animal ; Rats ; Rats, Sprague-Dawley

Objective To study the relationship between Couagen Ⅰ,MMP-2,TIMP-2 gene expression and atrial fibrosis during heart failure(HF)in dog.Methods Fourteen dogs were used and randomized into HF induced by ventricular tachypacing and control group.Burst atrial pacing was used to induce atrial fibrillation(AF).And the mRNA and protein level of collagen Ⅰ,MMP-2 and TIMP-2 were detected by RT-PCR and immunohistochemical technique.Tissue samples were stained with Mallory trichrome.Results Left ventricular ejection fraction (LVEF) decreased from (67.4? 6.0)% to (29.2?7.8)%,the inducible rate of AF(7/7 vs 2/7) and sustained AF(5/7 vs 0/7) increased and duration of AF stabeatrial fibrillation(SAF) [(462.12?181.43)s vs(0.57?0.57) s] prolonged significantly in HF group.Atrial fibrous tissue content and atrial size of HF group were significantly greater than the controls dogs(268.8% in lefe atria and 190.3% in right atria).The mRNA and protein level of collagen Ⅰ(56.2% and 132.2% in lefe atria,37.4% and 78.0% in right atria)and MMP-2 (100.0% and 115.7% in lefe atria,65.7% and 96.8% in right atria) increased evidently in both lefe atria and right atria,TIMP-2 mRNA decreased 46.3% in lefe atria and had no change in right atria and that its protein had no change in both atrium,whereas the ratio of MMP-2/ TIMP-2 of mRNA and protein increased markedly in both lefe atria (285.3% and 148.8%)and right atria (106.1% and 134.7%)of HF group.SAF had a positive correlation with fibrosis and the gene level of collagen Ⅰ in lefe atria,the ratio of MMP-2/TIMP-2 had a positive correlation with fibrosis and collagen Ⅰ gene level in lefe atria during HF.Conclusions The changes of collagen Ⅰ,MMP-2 and TIMP-2 gene expression appear to be a molecular mechanism of AF, and the molecular remodeling of collagen Ⅰ induced by regulation unbalance of MMP-2/TIMP-2 appears to be an important mechanism of atrial fibrosis during HF.

Aim To explore the mechanism of catechin in inhibiting Th2-driven inflammation responses of allergic asthma mice.Methods Female BALB/c mice were chosen to establish an allergic asthma model by sensitized and excited with OVA.Then,the administered group was intervened with catechin by series of dose (75,150,300 mg · kg-1).The proportion of peripheral leukocyte cells was analysed,and pathological changes in lung tissues were observed by HE dyeing.The levels of TSLP,IL-5,IL-13 in lung tissues and OVA specific IgE in mouse serum were measured using ELISA kit.NF-κB p65,IκB protein expression levels in lung tissues were investigated using Western blot,and NF-κB p65 nuclear translocation was detected with immunofluorescent technique.Results Catechin could not only effectively reduce the proportion of peripheral leukocyte cells,but relieve the levels of TSLP,Th2 inflammatory factor IL-5 and IL-13,and the activation of NF-κB signal pathway in lung tissues of allergic asthma mice.Conclusions Catechin can effectively relieve Th2 inflammation in allergic asthma mice induced by allergen OVA.The possible mechanism may be that it could reduce expression of TSLP by inhibiting the NF-κB signal.

Objective To observe the clinical efficacy and investigate its safety of esophageal achalasia with retrievable metal stents. Methods Thirty cases for esophageal achalasia in our hospital were selected, and they were randomly divided into stent group and balloon group. The former were treated by retrievable metal stent, and the latter were treated endoscopic balloon-dilating. The dysphagia grade, clinical symptom scores and the situation of esophageal in-spection before treatment,after 1 month and 6 months of treatment, and the common complications were compared be-tween two groups. Results (1)The rusults of two groups in dysphagia grade and clinical symptom scores after 1 month and 6 months' treatment were obviously lower than those before therapy(P<0.05).(2)Two groups patients in dysphagia grade and clinical symptom scores after 1 month's treatment had no statistical difference (P>0.05),and after six months' treatment, the stent group patients' dysphagia grade and clinical symptom scores were lower than balloon group (P<0.05). (3)After treatment, the maximum diameter of the esophagus of two groups patients were significantly lower than before treatment (P<0.05), and the narrowest diameter of the cardia were significantly higher than before treatment (P<0.05).(4)The rates of common complications such as hemorrhage, perforation, pain, stent group were lower than the balloon group(P<0.05),but reflux stent group was higher than the balloon group(P<0.05). Conclusion The treatment of esophageal achalasia with endoscopic retrievable metal stent is simple, safe and effective, compared with balloon-dilating,its long-term efficacy is more excellent,and the complications are more lighter and less.

OBJECTIVETo study the effect of ZGDHu-1 on proliferation, differentiation and apoptosis in SHI-1 human leukemia cell line and explore its possible mechanism. Methods SHI-1 cells were cultured with different concentration of ZGDHu-1 and for different time. The cell proliferation was analysed by cell counting, alive cell count, MTT assay and Brdu-ELISA. Cell apoptosis was analysed by morphology, DNA content, Annexin-V/PI and Hoechst 33258 labeling method. Cell differentiation were assayed by morphology,expression of CD11b,CD14 and CD64 and NBT reduction. The expressions of phosphorylated p38MAPK or STAT3 were analysed by flow cytometry.

RESULTSZGDHu-1 inhibited SHI-1 cell proliferation in a time and dose dependent manner, the IC50- 48 h and IC50- 72 h were 250 ng/ml and 85 ng/ml, respectively. The majority of SHI-1 cells were arrested in G2/M phase. 48h after treated with 200 ng/ml ZGDHu-1, and those in G2/M phase accounted for (48.4 +/- 2.1)%. The SHI-1 cells apoptosis was increased with a time- and does-dependent manner. The morphology of SHI-1 cells cultured with 2-50 ng/ml ZGDHu-1 for three days become more mature with higher NBT positivity and up-regulated expressions of CD11b,CD14 and CD64. The expression of phosphor-p38MAPK was increased and phosphor-STAT3 down-regulated by the treatment of ZGDHu-1.

CONCLUSIONZGDHu-1 can inhibit SHI-1 cell proliferation and induce the cell differentiation and apoptosis. The mechanism may associate with its up-regulation of phosphor-p38MAPK and down-regulation phosphor-STAT3.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Formamides ; pharmacology ; Heterocyclic Compounds, 1-Ring ; pharmacology ; Humans ; Leukemia ; pathology ; Phosphorylation ; STAT3 Transcription Factor ; biosynthesis ; p38 Mitogen-Activated Protein Kinases ; biosynthesis

To study the molecular mechanisms of nitric oxide donor sodium nitroprusside (SNP) -induced apoptosis in K562 human leukemia cell line, the different concentrations of SNP and different time of culture were used to treat K562 cell. At the same time, potassium ferricyamide (PFC) was used as control, blank was designed in experiment. Cell apoptosis was analysed by cell morphology, DNA agarose gel electrophoresis, DNA content, and annexin-V/PI labeling method. The TdT-mediated dUTP nick end labeling (TUNEL) assay was used to quantify in situ cell apoptosis. Reactive oxygen species (ROS) in cells and mitochondrial transmembrane potential (DeltaPsim) were labeled by dihydrorhodamin 123, 2', 7'-dichlorodihydrofluorescein diacetate and rhodamin 123/PI. bcl-2, bax, bad, p53 gene proteins and mitochondrial membrane protein were analysed by flow cytometry. The results showed that the K562 cell apoptosis was confirmed by typical cell morphology, DNA fragment, sub-G(1) phase, TUNEL and annexin-V/PI labeling. A majority of K562 cells were arrested in G(0)/G(1) phase. During the process of SNP-induced apoptosis in K562 cell, the mean fluorescence intensity of ROS in cells was significantly higher than those in blank and PFC control, while the DeltaPsim reduced. The expression of p53, bax, bad, Fas protein and mitochondrial membrane protein increased and bcl-2 protein decreased after SNP treatment. It is concluded that SNP induces K562 cell apoptosis through increasing ROS in cells, expressing the p53, bax, bad, Fas protein and mitochondrial membrane protein and decreasing bcl-2 protein, opening the mitochondrial permeability transition pore and reducing DeltaPsim. Furthermore, the Fas was activated during the apoptosis process.
Apoptosis ; drug effects ; Dose-Response Relationship, Drug ; Humans ; In Situ Nick-End Labeling ; K562 Cells ; Membrane Potential, Mitochondrial ; drug effects ; Nitric Oxide Donors ; pharmacology ; Nitroprusside ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Reactive Oxygen Species ; metabolism ; Tumor Suppressor Protein p53 ; metabolism ; bcl-2-Associated X Protein ; metabolism ; fas Receptor ; biosynthesis

OBJECTIVETo investigate the expression of occludin, ZO-1, MMP-2, and MMP-9 in cerebral microvasculature following Pulse Electromagnetic Field (PEMF) induced BBB permeability change.

METHODSSprague-Dawley rats were randomized into PEMF and sham exposed groups (n = 8). After exposure to PEMF at 0.5, 1, 3, 6, and 12 h, BBB permeability was measured by Evans-Blue extravasation. The expression of occludin, ZO-1, MMP-2, and MMP-9 were detected by real-time quantitative reverse transcriptase PCR and western blotting. MMP-2 and MMP-9 activity were detected by EnzChek gelatinase assay.

RESULTSCompared with the sham group, PEMF exposure led to increased permeability of the BBB to EB, which was prolonged after exposure. BBB permeability became progressively more severe, and recovered at 6 h. The gene and protein expression of occludin and ZO-1 were significantly decreased, while MMP-2 and MMP-9 expression were significantly increased after exposure to PEMF. All levels of expression recovered 12 h following PEMF.

CONCLUSIONChanges to BBB permeability were related to the alteration expression of tight junction proteins and matrix metalloproteinase after exposure to PEMF.

Animals ; Blood-Brain Barrier ; Electromagnetic Fields ; Male ; Matrix Metalloproteinases ; metabolism ; Proteins ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tight Junctions ; metabolism

This study was aimed to investigate the changes of reactive oxygen species and antioxidative capacity on nitric oxide induced apoptosis in HL-60 cells. By means of in vitro incubation of HL-60 cells with sodium nitroprusside (SNP), the growth inhibition was detected by MTT assay. Cell morphology was observed by transmission electronmicroscopy and light microscopy. The apoptosis was analyzed by DNA agarose gel electrophoresis, DNA content and Annexin-V/PI labeling method. Reactive oxygen species (ROS) labeled with dihydrorhodamin 123 in cells was determinated by flow cytometry. The SNP-treated cells were examined for glutathione (GSH) level and activity of catalase (CAT), glutathione S-transferase (GST) and glutathione peroxidase (GPX). The results indicated that SNP could inhibit HL-60 cell growth. Cell apoposis was confirmed by typical cell morphology, DNA fragment, sub-G(1) phase and Annexin-V/PI labeling method. HL-60 cell apoptosis was induced by SNP in a dosage- and time-dependent manner. After exposing to SNP at the concentration of 0.5 - 3.0 mmol/L for 48 hours, the mean fluorescence intensity of ROS in cells was significantly higher than those in groups control and potassium ferricyanide (PFC). During the apoptosis process, level of ROS in cells increased, levels of GSH, CAT, GPTand GPX decreased. The significant dose-effect relationship existed between the levels of ROS, CAT, GST, GPX and SNP dose. It is concluded that change of intracellular reactive oxygen species and antioxidative capacity are an important factors during the process of SNP-induced apoptosis in HL-60 cell.
Antioxidants ; metabolism ; Apoptosis ; drug effects ; Cell Nucleus ; drug effects ; ultrastructure ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; HL-60 Cells ; Humans ; Microscopy, Electron ; Nitric Oxide Donors ; pharmacology ; Nitroprusside ; pharmacology ; Reactive Oxygen Species ; metabolism

The diaphragm is one of the most important respiratory muscles in the human body,of which 60%-80%of the function of the respiratory muscle is played by the diaphragm.Mechanical ventilation is a common respiratory support method for severely ill patients in clinical.However,improper use of the ventilator can also lead to ventilator-induced diaphragm dysfunction(VIDD)while maintaining patients'respiratory function.The pathogenesis of VIDD is mainly induced by mechanical ventilation leading to diaphragmatic mitochondrial oxidative stress(MOS).In addition,the protein degradation of the ubiquitin-proteasome system(UPS),autophagy,changes in calpain activity,calcium,and cytokines also play important roles in the process of VIDD.In-depth understanding the pathological mechanism of VIDD is beneficial to prevent and treat VIDD.This article reviews the research progress of the potential etiology(improper use of ventilator,age,nutritional and metabolic status,coronary disease,drug factors)and pathogenesis(mitochondrial oxidative stress,autophagy of diaphragm cells,activation of calpain,and increase of intracellular calcium)of VIDD,so as to provide reference for the treatment of VIDD.