Alanine Transaminase ; metabolism ; Animals ; Liver ; drug effects ; Male ; Mice ; Plant Extracts ; pharmacology

BACKGROUNDIt has been widely observed that infants and young children can reossify large calvarial defects when they are younger than 2 years of age; afterwards, they lose this regenerative potential. Previous studies have implicated that the dura mater serves as a key regulator of calvarial regeneration. However, the molecular mechanism of calvarial reossification remains elusive.

METHODSIn order to identify the proteins that may participate in this process, we performed a proteome-wide comparison of the protein expression levels of immature and mature dura using 2D electrophoresis and MALDI-TOF mass spectrometry. The Western blotting was used to verify the results of the 2D electrophoresis/MALDI-TOF mass spectrometry.

RESULTSEleven proteins were found to express with significant differences in the immature and the mature dura. Among them, the emergence of vimentin, tropomyosin, beta-actin and gamma-actin were further confirmed by the Western blotting analysis.

CONCLUSIONThe proteins and proteomic profiles provide a better understanding of the molecular mechanism of calvarial regeneration.

Aging ; physiology ; Animals ; Blotting, Western ; Dura Mater ; chemistry ; Electrophoresis ; Mass Spectrometry ; Proteins ; analysis ; Proteomics ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the relationship between genetic polymorphisms of CACNA1C that encoded the a1c subunit of the L-type calcium channel and the efficacy of calcium channel blocker (CCB, Nifedipine extended release tablet/20 mg/d) in essential hypertension (EH) patients of Han Chinese in Wenzhou.

METHODSFor the enrolled 103 EH patients, Multiplex Polymerase Chain Reaction (Multi-PCR) and matrix assisted laser desorption ionization time of flight MS (MLDI-TOF MS) were performed to detect their genotypes (rs216008, rs1051375, rs2299661, rs10848683, rs215976), blood pressure (BP) after CCB monotherapy was compared among patients with different genotypes.

RESULTS(1) Blood pressure was significantly reduced in all patients post CCB (P < 0.05 vs. pre-CCB). (2) Diastolic blood pressure reduction was more significant in subjects with rs2299661 C/C genotype (wild genotype) than in subjects with rs2299661C/G and rs2299661G/G genotype (mutational genotype) [(12.46 ± 7.91) mm Hg (1 mm Hg = 0.133 kPa) vs. (7.22 ± 8.01) mm Hg and (5.93 ± 9.77) mm Hg, P < 0.05]. (3) Systolic blood pressure reduction was more significant in subjects with rs216008 C/C genotype (wild genotype) than in subjects with rs216008 C/T genotype (mutational genotype) [(20.60 ± 12.35) mm Hg vs. (13.62 ± 10.21) mm Hg, P < 0.05]. (4) Blood pressure reduction was similar between subjects with genotype of rs1051375, rs10848683 and rs215976.

CONCLUSIONEH patients with wild genotype of rs2299661 and rs216008 in CACNA1C are more likely to be responders of CCB monotherapy.

Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; Calcium Channel Blockers ; therapeutic use ; Calcium Channels, L-Type ; genetics ; Female ; Humans ; Hypertension ; drug therapy ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide

In this study, we investigated the mechanism of linoleic acid-stimulated increase in intracellular calcium concentration ([Ca(2+)](i)) in pancreatic islet β-cells. Pancreatic islet cells were primarily isolated from rats and cultured for the experiments. The cells were loaded with Fluo-3/AM, the indicator of [Ca(2+)](i), and the intensity of Fluo-3 was measured using confocal microscope. The islet β-cells were identified by immunocytochemical staining with insulin antibody after recording. The drugs were given by perfusion system. The results showed that linoleic acid (20 μmol/L) stimulated [Ca(2+)](i) increase with the first peak increase and the following plateau increase. Linoleic acid-stimulated [Ca(2+)](i) increase was partly inhibited by removal of extracellular calcium and by transient receptor potential (TRP) channel blocker, La(3+), and it was totally blocked by exhaustion of intracellular calcium stores and inhibition of phospholipase C. It is concluded that linoleic acid stimulates [Ca(2+)](i) increase in islet β-cells through both extracellular calcium influx via TRP channels and calcium release from intracellular calcium stores.
Animals ; Calcium ; metabolism ; Insulin-Secreting Cells ; cytology ; metabolism ; Linoleic Acid ; pharmacology ; Male ; Primary Cell Culture ; Rats ; Rats, Sprague-Dawley ; Transient Receptor Potential Channels ; antagonists & inhibitors

OBJECTIVETo explore the underlying mechanism of mesenchymal stem cells (MSCs) transfer induced cardiac function improvement in failing hearts.

METHODSCongestive heart failure (CHF) was induced in rats by cauterization of the heart wall. MSCs were cultured from autologous bone marrow and injected into the border zone and the remote myocardium 5 days after cauterization.

RESULTSTen weeks later, cardiomyocyte nucleus mitotic index, capillary density and expression of insulin-like growth factor 1 (IGF-1), hepatocyte growth factor (HGF) and vascular endothelial growth factor (VEGF) were significantly increased in the border zone and significantly reduced in the remote myocardium in CHF rats (all P<0.05 vs. sham). Besides cardiac function improvement and left ventricular remodeling attenuation evidenced by hemodynamic and echocardiographic examinations, expressions of IGF-1, HGF and VEGF in the remote myocardium and in the border zone were also significantly upregulated (P<0.05 or P<0.01 vs. CHF), and cardiomyocyte nucleus mitotic index as well as capillary density were significantly increased in CHF rats with MSCs (P<0.05 or P<0.01 vs. CHF). Moreover, collagen area was significantly reduced and myocardial area was significantly increased in the border zone in these rats too.

CONCLUSIONAutologous MSC implantation upregulated expressions of growth factors enhanced cardioangiogenesis which might be the underlying mechanisms for improved cardiac function and attenuated left ventricular remodeling induced by MSCs transplantation in failing rat myocardium.

Animals ; Disease Models, Animal ; Heart Failure ; metabolism ; therapy ; Hepatocyte Growth Factor ; metabolism ; Insulin-Like Growth Factor I ; metabolism ; Male ; Mesenchymal Stem Cell Transplantation ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Transplantation, Autologous ; Vascular Endothelial Growth Factor A ; metabolism ; Ventricular Remodeling

OBJECTIVETo study the role of carbamyl phosphate I (CPS-I)and ornithine transcarbamoylase (OCT) levels in cirrhosis patients with and without hepatic encephalopathy, and to analyze the correlations between CPS-Iand OCT with the development of hepatic encephalopathy.

METHODSCPS-I, OCT, plasma ammonia and liver function of 95 cirrhosis patients with hepatic encephalopathy and 25 cirrhosis patients without hepatic encephalopathy in our hospital from January 2008 to December 2009 were analyzed. 60 healthy controls were recruited in the control group. The differences of serum CPS-I, OCT levels among the cirrhosis patients with and without hepatic encephalopathy and the healthy controls were analyzed; the correlations of CPS-I, OCT levels with plasma ammonia and total protein in cirrhosis patients,and the correlations of CPS-I, OCT levels with Child-Pugh classification of cirrhosis symptom severity in cirrhosis were analyzed. the clinical characteristics between patients who had HE and no HE with chi-square tests were compared. Comparisons of CPS-I, OCT levels across patients based on the Child-Pugh classification were performed with One-Way ANOVA and Student-Newman-Keuls, correlation of CPS-I, OCT with other indicators were performed with Pearson correlation analysis.

RESULTSSerum CPS-I and OCT levels in cirrhosis patients with hepatic encephalopathy were (143.3+/-48.5) U/L, (297.0+/-102.6) is multiplied by 10 U/L, which were lower than that in cirrhosis patients without hepatic encephalopathy (180.3+/-51.5) U/L, (351.8+/-109.0) is multiplied by 10 U/L (t = 2.53, t = 2.78, P < 0.01). Compared with healthy controls, serum CPS-I and OCT levels in cirrhosis patients with and without hepatic encephalopathy were all lower (t = 3.21, t = 4.16, t = 2.12, t = 3.15, P < 0.05). CPS-I was correlated with OCT, (r = 0.946, P < 0.05); CPS-I and OCT were negatively correlated with ALT and AST (r = -0.284, r = -0.239, r = -0.303, r = -0.322, P < 0.05). Additionally, CPS-I and OCT levels were negatively correlated with the Child-Pugh classification in Cirrhosis (F = 10.13, F = 20.28, P < 0.01).

CONCLUSIONThe serum CPS-I and COT levels were important factors affecting plasma ammonia in patients with cirrhosis and played an important role in the development of hepatic encephalopathy.

Adult ; Ammonia ; blood ; Carbamoyl-Phosphate Synthase (Ammonia) ; metabolism ; Case-Control Studies ; Female ; Hepatic Encephalopathy ; blood ; enzymology ; Humans ; Male ; Middle Aged ; Ornithine Carbamoyltransferase ; metabolism

OBJECTIVETo investigate the effect on early treatment with vacuum-assisted closure(VAC) to wound healing of acute explosion injury in pigs, and provide a new way for early treatment of battle wounds.

METHODSEight healthy 3-month Landrace pigs of both sexes with the body mass of (50 +/- 5) kg were selected in the study. Sixteen battle wounds were made by explosion of same type detonator (pattern number: 660929F48840-55, included DDNP 0.3 g, RDX 0.7 g) in hibateral skin of buttock of 8 pigs, which were divided into experimental group and control group (pair wounds of left and right). The raw sufaces were thorough debrided at 3 h after exposure, according to the characteristics of treatment on the battlefield, experimental group was treated with VAC under the pressure of (-50 +/- 5) Kpa after debridement and sterilization and control group was treated with routine dry sterile gauze draping. Results of bacteriology (bacterial counts and the proportion of G+ bacteria) and pathology (HE stain and Masson stain) were detected at every wound before and after treatment.

RESULTSAt the 3 days after treatment,the bacterial number in the experimental group was [(7.82 +/- 0.55) x 10(4) ] CFU/g, in control group was [(1.07 +/- 0.14) x 10(6)] CFU/g. There was significant difference between two groups. The proportion of G+ bacteria in experimental group was significantly increased. The raw surface in experimental group was clean with affluent and neoformative granulation tissue, blood vessels and collagen, necrotic tissue decreased obviously by pathological observation.

CONCLUSIONVAC could reduce the quantity of bacteria, improve the proportion of G+ bacteria, and promote the formation of granulation tissue and the healing of wound. The VAC for the treatment of battle wounds has a positive effect.

Animals ; Colony Count, Microbial ; Explosions ; Female ; Male ; Negative-Pressure Wound Therapy ; methods ; Soft Tissue Injuries ; etiology ; microbiology ; pathology ; surgery ; Staining and Labeling ; Swine ; Time Factors

OBJECTIVETo investigate the effect of ghrelin on duodenal migrating myoelectric complex (MMC) in rats with chronic renal failure (CRF).

METHODSThirty healthy male SD rats were randomly assigned into sham-operated group (n=6) and CRF group (n=24), and the latter group was divided into 4 subgroups according to ghrelin doses administered with or without pretreatment with the receptor antagonist D-Lys(3)-GHRP-6. After a 18-h fasting, the rats with or without pretreatment with D-Lys(3)-GHRP-6 were given subcutaneous injections of ghrelin at different doses to observe the changes in duodenal MMC recorded using a multi lead physiological recording system.

RESULTSGhrelin significantly increased the MMC cycle duration and dose-dependently enhanced the frequency, amplitude and percentage of phase III MMC cycle. This effect was inhibited by the pretreatment with ghrelin receptor antagonist D-Lys(3)-GHRP-6.

CONCLUSIONGhrelin can promote gastrointestinal motilities of rats with CRF, and the receptor of ghrelin can regulate the activity of MMC.

Animals ; Duodenum ; drug effects ; Gastrointestinal Motility ; drug effects ; Ghrelin ; pharmacology ; Kidney Failure, Chronic ; physiopathology ; Male ; Myoelectric Complex, Migrating ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the origin of oxidative stress induced by angiotensin II (AngII) in human mesangial cells and the role of reactive oxygen species (ROS) in AngII-induced monocyte chemoattractant protein-1 (MCP-1) expression.

METHODSMCP-1 expression was determined by real time RT-PCR. ROS production was measured by DCFDA fluorescence. Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity was examined by lucigenin chemiluminescence. p47phox and p67phox translocation was assayed by Western blot. Twenty-four male mice were randomly divided into three groups: the control, the AngIIinfusion [AngII 400 ng/(kg.min)], and the apocynin treatment. AngII was infused by subcutaneously osmotic minipump for 14 days. Urinary albumin and 8-isoprostane excretion were measured by ELISA.

RESULTSIn cultured human mesangial cells, AngII induced the MCP-1 expression in a dose-dependent manner with 3.56 fold increase as compared with the control. AngII increased intracellular ROS production as early as 3 min with the peak at 60 min and was in a time and dose-dependent. Incubation with different dosages of AngII (1 nmol/L, 10 nmol/L, and 100 nmol/L AngII) for 60 min, ROS production increased at 1.82, 2.92, and 4.08 folds respectively. AngII-induced ROS generation was sensitive to diphenyleneiodonium sulfate (DPI, 10 micromol/L) and apocynin (500 micromol/L), two structurally distinct NADPH oxidase inhibitors. In contrast, inhibitors of other oxidant-producing enzymes, including the mitochondrial complex Iinhibitor rotenone, the xanthine oxidase inhibitor allopurinol, the cyclooxygenase inhibitor indomethacin, the lipoxygenase inhibitor nordihydroguiaretic acid, the cytochrome P450 oxygenase inhibitor ketoconazole and the nitric oxide synthase inhibitor G-nitro-L-arginine methyl ester were without an effect. AngII-induced ROS generation was inhibited by the AT1 antagonist losartan (10 micromol/L) but not the AT2 antagonist PD123319 (10 micromol/L). AngII treatment induced translocation of cytosolic of p47phox and p67phox to the membrane. The antioxidants almost abolished AngII-induced MCP-1 expression. AngII infusion increased urinary and p67 translocation by 2.69-, 2.97-, and 2.67-fold, respectively.

CONCLUSIONSNADPH oxidase-derived ROS is involved in AngII-induced MCP-1 expression. Inhibition of NADPH oxidase alleviates AngII-induced renal injury.

Acetophenones ; pharmacology ; Angiotensin II ; administration & dosage ; pharmacology ; Angiotensin II Type 1 Receptor Blockers ; pharmacology ; Animals ; Cells, Cultured ; Chemokine CCL2 ; metabolism ; Dose-Response Relationship, Drug ; Humans ; Losartan ; pharmacology ; Male ; Mesangial Cells ; metabolism ; Mice ; Mice, Inbred C57BL ; NADPH Oxidases ; antagonists & inhibitors ; metabolism ; Onium Compounds ; pharmacology ; Oxidative Stress ; Phosphoproteins ; metabolism ; Protein Transport ; Random Allocation ; Reactive Oxygen Species ; metabolism

This study was aimed to quantitatively analyze the mRNA level of bcr-abl fusion gene in K562/A02 cell line by real-time quantitative reverse transcriptase polymerase chain reaction (RQ-RT-PCR) technique. After being cultured for a period of time, the K562/A02 cell line was collected and RNA was extracted using TRIzoL kit. The real-time quantitative reverse transcriptase polymerase chain reaction technology was used to detect the level of bcr-abl fusion gene and internal reference abl gene. The results showed that a fine reproducibility was obtained between 10(7) and 10(3) copies/ml, reproducible sensitivity of RQ-RT-PCR was 10(-5). The expression of bcr-abl fusion gene in K562/A02 cells was higher and the level of bcr-abl mRNA was more than 100% in K562/A02 cells. It is concluded that RQ-RT-PCR is a reliable, sensitive and reproducible method for detecting mRNA level of bcr-abl fusion gene, which may be useful in monitoring the chronic myeloid leukemia.
Fusion Proteins, bcr-abl ; genetics ; Humans ; K562 Cells ; RNA, Messenger ; genetics ; Real-Time Polymerase Chain Reaction ; methods ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity