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Acute kidney injury induced by renal ischemia-reperfusion (IR) is a prominent risk factor in the development towards renal fibrosis. Ras-related C3 botulinum toxin substrate 1(Rac1) has been involved in the pathophysiology of fibrotic disorders. But the role of Rac1 in the pathogenesis of IR-induced renal fibrosis is still unknown. Here, we examined the effects of NSC23766, an inhibitor of Rac1, on the progression of renal fibrosis after IR injury. In mice, IR induced Rac1 activation in kidneys. Rac1 inhibition alleviated renal damage and dysfunction. Mice treated with NSC23766 displayed diminished collagen area in the kidneys compared with IR controls, which was associated with reduction of extracellular matrix (ECM) proteins and α-SMA. Furthermore, Rac1 inhibition reduced profibrotic molecules levels in the kidneys of mice with IR. Finally, Rac1 inhibition impaired the accumulation of bone marrow-derived M2 macrophages and the transition of M2 macrophages to myofibroblasts. In cultured mouse monocytes, IL-4 treatment activated Rac1, which was abrogated by NSC23766. Moreover, application with IL-4 induced polarization of monocytes to M2 phenotype and increased the levels of ECM proteins and α-SMA, which was abolished by NSC23766. In summary, Rac1 plays a crucial role in the pathogenesis of renal fibrosis after IR via regulation of expressions of profibrotic molecules, bone-marrow derived M2 macrophages recruitment, and M2 macrophages-myofibroblasts transition.

A ZnO and carbon nanoparticle (NP) mixture has been prepared. The fluorescence color of the mixture changes from yellow to blue when the pH value of the ambient is smaller than 5.5, which accords well with the pH standard value of acid rain (

Hypoxia promotes the progression of lung cancer cells. Unfortunately, anesthetic technique might aggravate hypoxia of lung cancer cells. Sevoflurane is a commonly used anesthetic. Its effect on hypoxia-induced aggressiveness of lung cancer cells remains unknown. The aim of the study is to investigate the effects of sevoflurane on hypoxia-induced growth and metastasis of lung cancer cells. As hypoxia-inducible factor-1α (HIF-1α) plays a pivotal role in mediating the adaptation and tolerance of cancer cells under hypoxic microenvironment, the role of HIF-1α in the effect of sevoflurane on hypoxia-induced growth and metastasis has also been elucidated. A549 cells were treated with normoxia, hypoxia, co-treatment of sevoflurane and hypoxia, and dimethyloxaloylglycine (DMOG, a HIF-1α agonist) for 4 h, respectively. MTT assay and colony formation assay were used to evaluate cell growth. Transwell assay was performed to detect invasion and migration ability. The protein level of HIF-1α, X-linked inhibitor of apoptosis protein (XIAP), survivin, fascin, heparanase (HPA), and p38 MAPK were determined by Western blotting. Hypoxia enhanced proliferation and metastatic potential of cells. Sevoflurane could suppress hypoxia-induced growth and metastasis ability of cells. Furthermore, HIF-1α, XIAP, survivin, fascin and HPA were down-regulated significantly by the co-treatment of sevoflurane and hypoxia as compared to hypoxia treatment. DMOG abolished the inhibiting effects of sevoflurane on hypoxia-induced growth and metastasis ability of cells. In addition, sevoflurane partly reversed the increase of p38 MAPK activity that was induced by hypoxia. Sevoflurane could suppress hypoxia-induced growth and metastasis of lung cancer cells, which might be associated with modulating HIF-1α and its down-stream genes. Moreover, p38 MAPK signaling pathway was involved in the regulation of HIF-1α by sevoflurane.

The interferon-γ-inducible lysosomal thiol reductase (GILT) has been demonstrated to play an important role in the processing and presentation of MHC class II restricted antigen (Ag) by catalyzing disulfide bond reduction. In this study, we cloned a GILT gene homolog from goldfish (designated gGILT), a kind of precious freshwater fish with high market value. The open reading frame of gGILT consists of 756 bases encoding a protein of 251 amino acids with an estimated molecular mass of 27.8 kDa and a theoretical isoelectric point of 5.24. The deduced protein possesses the typical structural features of known GILT proteins, including an active-site motif, a GILT signature sequence, and 10 conserved cysteines. RT-PCR results showed that gGILT and gIFN-γ (goldfish IFN-γ) mRNA were expressed in a tissue-specific manner and obviously up-regulated in splenocytes and the cells from head kidney after induction with LPS. Recombinant gGILT fused with His6 tag was efficiently expressed in Escherichia coli BL21 (DE3) and purified by Ni-NTA affinity chromatography. Further study revealed that gGILT was capable of catalyzing the reduction of the interchain disulfide bonds from intact IgG. This study shows that gGILT may be involved in the immune response to bacteria challenge and maintain first line of innate immune defense at basal level in goldfish. It also provides the basis for investigating on the role of GILT using goldfish as an animal model.

The tumor necrosis factor superfamily (TNFSF) proteins are cytokines involved in many biological processes. In this study, the TNF superfamily member 14 (TNFSF14, LIGHT) has been isolated from zebrafish Danio rerio (designated zLIGHT). The full-length open reading frame (ORF) of zLIGHT cDNA consists of 708 bp encoding a protein of 235 amino acids. The zLIGHT open reading frame (ORF) genomic sequence consists of three introns and four exons, is about 9.9 kb in size. Real-time quantitative PCR (qPCR) analysis suggested that zLIGHT was predominantly expressed in zebrafish spleen. The soluble LIGHT (zsLIGHT) had been cloned into the pSUMO vector, SDS-PAGE and Western blotting analysis confirmed that the recombinant protein SUMO-zsLIGHT was efficiently expressed in Escherichia coli BL21 (DE3). Laser scanning confocal microscopy analysis showed that SUMO-zsLIGHT could bind to its receptors on T cells. LIGHT is involved in many important biological effects, including up-regulating proinflammatory chemokines, cytokines, inducing cell death, apoptosis, and enhancing T cell survival. Zebrafish may conduct as a model animal for further research on LIGHT.

Objectives: It was the aim of this study to investigate the effect of ZP123 on prolonged ventricular fibrillation (VF) in swine. Methods: VF was electrically induced in 20 pigs. The animals randomly received either ZP123 or saline control infusion before VF. After 8 min of untreated VF, cardiopulmonary resuscitation and biphasic defibrillation shocks were applied. VF mean frequency (VFmf) and mean amplitude (VFma), hemodynamics, outcome of defibrillation and the rate of return of spontaneous circulation (ROSC) were analyzed. Results: Compared with the control group, VFmf was higher but VFma lower during the 8 min of VF in the drug group (11.8 ± 2.1 vs. 10.4 ± 2.0 Hz and 0.24 ± 0.10 vs. 0.31 ± 0.16 mV, respectively; p < 0.05). Hemodynamic variables in the 2 groups were comparable (p > 0.05). The defibrillation threshold was lower and the rate of successful defibrillation was higher in the drug group compared with the control group (92.2 ± 26.4 vs. 133.3 ± 28.9 J and 90 vs. 30%, respectively; p < 0.05). The rate of ROSC was not different between the 2 groups (40 vs. 30%; p > 0.05). Conclusion: In prolonged VF, ZP123 could decrease the defibrillation threshold and improve the rate of successful defibrillation. However, it could not improve the rate of ROSC – which may be due to its side effect of decreasing VFma.

A simple and sensitive method based on the derivatization of 20 amino acids with 1,2-benzo-3,4-dihydrocarbazole-9- ethyl chloroformate (BCEOC) as derivatization reagent on a reversed-phase Hypersil BDS C 18 column (4.6 mm × 200 mm, 5 μm) with a gradient elution followed by high performance liquid chromatography (HPLC) via fluorescence detection at 390 nm (excitation wavelength of 333 nm) and tandem mass spectrometric identification has been developed. Optimum derivatization, giving the corresponding stable fluorescent derivatives, was obtained by reacting amino acids with BCEOC at 40 ° for 10 min in borate buffer (pH 9.0) with four times excess of molar reagent. The linear range of 20 amino acids were 51.6 fmol–105.6 pmol, all correlation coefficients >0.9995, detection limits were 6.3–177.6 fmol (at signal to noise 3:1, S/N =3:1). The identification of amino acid derivatives was carried out by post-column tandem mass spectrometry with electrospray ion (ESI) source, and the MS/MS cleavage mode of representative tyrosine derivative was analyzed. Under all the above optimum experimental conditions, the contents of 20 amino acids in rat serum of 3 groups (A: Quiet, B: At exercising exhaust, C: 12 h after exercising exhaust) were determined. It was indicated that the contents of 20 amino acids in group B were obviously higher than those in group A, and the contents of amino acids of group C and group A were almost the same. The established method exhibited high sensitivity and excellent reproducibility, and provided a new technology for the determination of amino acids in rat serum.

To study the changes of cardiovascular regulating factors in rats during recovery of aerobic exhaustive exercise.Sixty male Wistar rats were randomly divided into control group, 1 h-exercise group, 3 h-exercise group, exhausted group, 2 h-recovery group and 12 h-recovery group. The rats were killed at corresponding times for each group after an 8-week-long treadmill training, and the levels of NO, ET, ANP and TXB2 in plasma were measured in each group.NO/ET ratio of 1 h-exercise group was significantly higher than that in control group (P0.01), while it was significantly decreased in 3 h-exercise group and exhausted group (P0.05). ANP contents in rat plasma were significantly higher in 3 h-exercise group, exhausted group and 2 h-recovery group than that in control group (P0.05 or P0.01). The concentration of TXB2 in plasma was significantly increased in 3 h-exercise group, exhausted group and 2 h-recovery group (P0.05).Changes in cardiovascular regulating factors after exhaustive exercise may lead to deficiency of coronary circulation blood/oxygen supply, which may cause exercise-induced fatigue.

The aim of this study is to investigate the dynamic alterations of cardiac connexin 43 (Cx43), matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-2 (TIMP-2) in the setting of different ventricular fibrillation (VF) duration. In this study, thirty-two dogs were randomly divided into sham control group, 8-min VF group, 12-min VF group, and 30-min VF group. Cx43 and phosphorylated Cx43 (p-Cx43) in tissues were detected by western blot and immunofluorescence analysis. MMP-2 and TIMP-2 were detected by western blot and immunohistochemistry analysis. The results showed that Cx43 levels in three VF groups were significantly decreased compared with sham control group. p-Cx43 levels in 12-min and 30-min VF groups were significantly reduced compared with sham control group. The ratio of p-Cx43/Cx43 was also decreased in VF groups. Compared with sham controls, no significant difference was observed between the sham control group and 8-min VF group in MMP-2 level, but MMP-2 level increased in 12-min and 30-min VF groups. The ratios of MMP-2/TIMP-2 were higher in VF groups, and were correlated with the duration of VF. A remarkable correlation was observed between the ratio of p-Cx43/Cx43 and MMP-2/TIMP-2 (r = −0.93, P

Sevoflurane, an inhalational anesthetic, and cisplatin (DDP)-based chemotherapy have been widely used during lung cancer surgery. However, the effect of sevoflurane on the sensitivity of lung cancer cells to DDP chemotherapy remains unclear. In this study, the effects of combined treatment with sevoflurane and cisplatin on the growth and invasion of human lung adenocarcinoma A549 cell line have been investigated. The underlying mechanism has also been explored. In our experiment, A549 cells were treated with 2.5% sevoflurane, 10 μmol/L DDP, or the co-treatment of sevoflurane and DDP for 4 h, respectively. Cell proliferation was evaluated by the MTT assay and colony formation assay. Apoptosis was assessed by flow cytometry. Cell invasion was detected by Transwell assay. The expressions of X-linked inhibitor of apoptosis protein (XIAP), Survivin, matrix metalloproteinase (MMP)-2 and MMP-9 were determined by western blotting. Our results showed that sevoflurane combined with DDP resulted in a more pronounced inhibition of tumor cells growth and invasion as compared with either drug alone. Besides, XIAP, Survivin, MMP-2, and MMP-9 were downregulated more significantly by the co-treatment of the two drugs as compared to sevoflurane treatment or DDP treatment alone. Taken together, the growth-inhibitory and invasion-inhibitory synergy between sevoflurane and DDP in human adenocarcinoma A549 cell line was found in this study. Furthermore, we showed that the growth-inhibitory synergy between sevoflurane and DDP might be associated with the downregulation of XIAP and Survivin, and the invasion-inhibitory synergy between sevoflurane and DDP might be involved in the downregulation of MMP-2 and MMP-9.

To explore the influence of different strength intermittent treadmill training of growth period rats on the bone metabolism, so as to provide the training intensity of teenagers to set theory support.Select 70 male four weeks Wistar rats according to body weight randomly divided into seven groups (n = 10): the control group and the exercise group. According to the VO2max the exercise group was divided into 6 groups: 65%, 70%, 75%, 80%, 85% and 90% group. Nine weeks treadmill training, training six days a week, each group of training three times, each time not less than 10min, the interval was 30 min. The last movement after 24 h, took the femur and blood to measured the bone mineral density (BMD), bone mass (BMC) and alkaline phosphatase (AKP), resist tartaric acid acidic phosphatase (Str-ACP).1. The femoral BMD (0.1393 +/- 0.0031), BMC (0.4525 +/- 0.0335) of 70% group were significantly higher than those in the control group (BMD: 0.1200 +/- 0.0095, BMC: 0.3238 +/- 0.0485) and the other sports group (65% BMD:0.1339 +/- 0.0062, BMC: 0.4058 +/- 0.0492, 75% BMD: 0.1296 +/- 0.0015, BMC: 0.3869 +/- 0.0254, 80% BMD: 0.1223 +/- 0.0082, BMC: 0.3454 +/- 0.0483, 85% BMD: 0.1250 +/- 0.0044, BMC: 0.3731 +/- 0.0381, 90% BMD: 0.1171 +/- 0.0047, BMC: 0.3051 +/- 0.0286) (P0.05), the femoral BMD, BMC of 90% group were lower than those of the control group, the other in the exercise group were higher than those in the control group; 2. Serum AKP in the exercise group were significantly higher than those in the control group, the group of 65% (41.015 +/- 2.114), 70% (46.035 +/- 2.611), 75% (43.834 +/- 3.155), and 80% (38.043 +/- 4.073) were very significantly higher than those in the control group (26.875 +/- 1.188) (P0.01); 70% group and 75% group were significantly higher than those in the 80% group , 85% group and 90% group, while 70% group serum AKP level were significantly higher than those in 65% group (P0.05), it showed that 70% of the VO2 max training intensity of osteoblasts was most active. The serum Str-ACP of exercise group were significantly higher than those in the control group, along with the increase of the training intensity, serum Str-ACP level was rising and the group of 80% (22.430 +/- 1.591), 85% (23.990 +/- 1.870), and 90% (28.009 +/- 1.839) serum of Str-ACP were significantly higher than those in the group of 65% (18.503 +/- 2.429), 70% (16.447 +/- 2.120) and 75%(17.769 +/- 1.642) ( P0.05), the group of 90% serum Str-ACP were significantly higher than those in the group of 80% and 85% (P0.05).The training of 70% of the VO2max, moderate intensity intermittent running, make the growth period rat bone mass and bone mineral density to increase obviously.

To investigate the effect of the "compound extract of Chinese medicine" on free radical metabolism and antioxidant enzyme systems of the rat brain.70 Wistar mice were randomly divided into two groups (n = 35): normal control group (N), taking medicine group (M). After a week of feeding, M group was taking medicine for 8 weeks. After 9 weeks, killed the two groups when they were at the rest state, the immediate ends of the fixed load, the immediate ends of exhaustive exercise and 12, 24 hour ends of exhaustive exercise, respectively. And the activity of malonaldehyde (MDA), glutathione peroxidase (GSH-PX), glutathione (GSH), superoxide dismutase (SOD), total anti oxidation capacity (T-AOC) of the rat brain were measured.At the five states, the content of MDA in M group was lower than that in N group in different degree, the activity of GSH-PX, GSH, SOD, T-AOC in M group were higher than those in N group in different degree.The "Compound extract of Chinese medicine" can reduce the MDA level of the rat brain and improve the enzyme activity of GSH-PX, GSH, SOD, T-AOC.

To observe the effect of Chinese herbal compound on variance of neurotransmitters in rat telencephalon and to further discuss the mechanism underlying Chinese herbal compound in improving exercise capacity and promoting recovery from exercise-induced fatigue.64 rats (8 week old) were randomly divided into medicine group (MG) and control group (CG). Chinese herbal compound was administered to rats of MG for 8 weeks. 8 weeks later, every group was divided into 4 subgroups and all were killed at different time point separately, and then neurotransmitter in rat brain was tested.The exhaustion time of MG was significantly longer than that in CG (P0.01). In rest conditions, glutamic acid (GLU) of MG was significantly higher than that in CG (P0.01), while, there were no significant differences between MG and CG in other indexes. After fixed quantitative load exercise, the content of 5-hydroxytryptamineZZ(5-HT), 5-hydroindole acetic (5-HIAA), gamma-aminobutyric acid (GABA), Dopamine (DA) and 5-HT/5-HIAA were significantly lower than those in CG, while, GLU, GLU/GABA and DA/5-HT were significantly higher than those in CG. Compared with CG, exhaustion significantly (P0.05) decreased 5-HT, GABA and 5-HT/5-HIAA, and significantly (P0.05) increased GLU, DA/5-HT and GLU/GABA level in MG. 12 h after exhaustion, in contrast to CG, level of 5-HT and 5-HT/5-HIAA in MG were significantly (P0.01) lower while GLU, DA, GABA and DA/5-HT were significantly (P0.01) higher.During exhaustion exercise, Chinese herbal compound demonstrated strong inhibiting effect on synthesis of 5-HT, 5-HIAA, DA, GABA and promoting effect on GLU synthesis, this had been confirmed by the combined effect, including increase of excitatory transmitter and excitability of central nervous system and the prolongation of exhaustion time and promoting recovery from fatigue.

This study aimed to determine whether (a) there was an imbalance between matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1) after cardiopulmonary resuscitation (CPR) in a canine model of prolonged ventricular fibrillation (VF); (b) with the duration of VF, the degree of the imbalance would be greater; and (c) there was a relationship between the level of MMP-9 or TIMP-1 and the cardiac function.Ventricular fibrillation was electrically induced in 24 dogs. The animals were randomly divided into 3 groups (sham control, n = 8; 8-minute VF, n = 8; 12-minute VF, n = 8). Echocardiographic measurement and hemodynamic variables were recorded before VF and after return of spontaneous circulation. Tissue inhibitor of metalloproteinase 1 (TIMP-1) and MMP-9 were analyzed by Western blot and immunohistochemistry. Compared with sham controls, dogs under VF and CPR showed significantly decreased level of TIMP-1 (P.001), and with the duration of VF, the level of TIMP-1 declined (P.01). The level of MMP-9 did not achieve statistical significance in the 3 groups (P.05); however, they were higher in VF and longer duration VF groups. The ratios of TIMP-1/MMP-9 were lower in VF groups (P.05). There was a negative correlation between TIMP-1 and left atrium dimension and left ventricular diastolic dimensions (r = -0.83 and r = -0.96, respectively; P.01) and a positive correlation between TIMP-1 and left ventricular ejection fraction (r = 0.85; P.01).There was an imbalance between TIMP-1 and MMP-9 after CPR. It may partly contribute to the postresuscitation cardiac dysfunction.

To observe changes in connexin 43 (Cx43) after ventricular fibrillation (VF) and the effects of rotigaptide (ZP123) on Cx43.Thirty domestic pigs were randomly assigned to three groups (10 in each group): sham group, model group and ZP123 group. VF was induced by an 80 V AC transthoracic shock for 5 seconds with the use of subcutaneous needles. Before the induction of VF, animals in ZP123 group were administered with ZP123 (1 μg/kg bolus+10 μg×kg(-1)×h(-1) dissolved in 50 ml normal saline and pumped for 15 minutes ). Those in model group received 50 ml normal saline pumped for 15 minutes. For pigs in sham group VF was not induced and no fluid was given. After 8 minutes of VF, animals were euthanized and myocardial tissues were harvested along the long axis of each left ventricular free wall. Immunofluorescence combined with laser scanning confocal microscope was used to detect the distribution of Cx43. Western blotting was used for quantitative determination of Cx43 protein expression.Immunofluorescence signals for Cx43 in sham group were strong and regularly distributed. In model group, Cx43 signals were weak and distributed in heterogeneity, while in ZP123 group, Cx43 signals were enhanced and their distribution were much more orderly. Compared with sham group, the percentage area and the optical densities (A value) of Cx43 fluorescence signals and Cx43 protein expression were significantly decreased in model group [the percentage area: (0.64±0.36)% vs.(1.27±0.19)%, A value: 15 201± 2 613 vs. 30 634±4 975, Cx43 protein expression: 0.72±0.08 vs. 0.97±0.07, all P0.05]. The level of Cx43 expression in ZP 123 group [the percentage area (0.96±0.16)%, A value 22 100±4 404, Cx43 protein expression 0.82±0.04] was much higher than model group (all P0.05).During VF, down-regulation of myocardial Cx43 expression occurred, which could be attenuated by administration of ZP123.

To study the immunogenicity of Bordetella avium OMP, ultrasonic dispersion, TritonX-100 technique were used to extract Bordetella avium OMP. Its content was determined by Bradford method and it was detected by SDS-PAGE. Then OMP immunizing antigen was prepared and 1-day chicken was vaccinated by hypodermic inoculation, with the immunizing does of 0.3mL (OMP90microg), 0.5mL (OMP150microg), 0.8mL (OMP240microg), respectively. Results showed that content of Bordetella avium OMP is 300 g/mL, the best immunizing does is 0.5mL each one and Chickens can be protected against Bordetella avium at fetal dose if antibody titer is over 1:2(8). We can see from the antibody level detected by indirect ELISA that antibody can last long enough to help chickens pass susceptible period, so OMP has good immunogenicity. Results of this study lay good foundation for the development of monoclonal antibody to OMP, rapid diagnosis kit and subunit vaccine.

Cancer patients have an increased risk of thrombosis after operation because of a hypercoagulable status. Therefore, anticoagulant treatment is necessary for patients with hypercoagulability during perioperative period. This study was to investigate the effect of acute hypervolemic hemodilution (AHHD) with 6% hydroxyethyl starch (HES), or 4% succinylated gelatin (GEL), or lactated Ringer's (RL) solution before operation on the hypercoagulable status and the occurrence of deep venous thrombosis (DVT) of patients with colon cancer.Sixty colon cancer patients with hypercoagulable status underwent operation were randomized into HES, GEL, and RL groups; each group contained 20 patients. The patients were infused with HES, GEL, or RL solution respectively at a dose of 15 ml/kg within 30 min before operation. Preoperative coagulation function was assessed by thrombelastography (TEG). DVT was diagnosed by the color Doppler ultrasonic system.Acute hypervolemic hemodilution with HES solution led to a significant decrease of coagulation index (CI) at 30 min and 2 h after starting operation and 1 h after operation as compared with the prehemodilution value (P0.01). At 1 h after operation, CI was significantly lower in HES group than in GEL group (P0.05). At 30 min and 2 h after starting operation and 1 h after operation, CI was significantly lower in HES group than in RL group (P0.05). Hemodilution with GEL solution lessened CI at 30 min and 2 h after starting operation significantly as compared with the prehemodilution value (P0.01). At 30 min and 2 h after starting operation, CI was significantly lower in GEL group than in RL group (P0.05). After operation, DVT in occurred 2 (10%) patients in HES group, 3 (15%) in GEL group, and 10 (50%) in RL group (P0.05).Acute hypervolemic hemodilution with HES solution and GEL solution can alleviate the hypercoagulability of colon cancer patients during perioperative period and decrease the occurrence of DVT.