OBJECTIVETo explore roles of extracellular signal-regulated kinase (ERK) 1/2, p38 mitogen activated protein kinase (p38 MAPK) and nuclear factor (NF) -KB in expression of inducible nitric oxide synthase (iNOS) in rat alveolar macrophages induced by high mobility group box 1 (HMGB1 ).

METHODSPrimary rat alveolar macrophages (PRAMs) cultured in vitro were incubated with PD98059 ( inhibitor against ERK), SB203580 (inhibitor against p38 MAPK) , PDTC (inhibitor against NF-kappaB), or PD98059 plus SB203580 for 2 hours, respectively. HMGB1 was added into the cultures and incubated with cells for 6 hours. Total RNA of PRAMs was extracted and iNOS mRNA expression was semi-quantified with reverse transcription-polymerase chain reaction ( RT-PCR). Greiss reaction was applied to determine nitrite/nitrate (NO2-/NO3- ) concentration in PRAMs culture supernatants.

RESULTSExpression of iNOS mRNA and NO production in PRAMs culture supernatants were down-regulated by inhibition of ERK or p38 MAPK by PD98059 or SB203580, respectively (P <0. 05). Moreover, inhibition of iNOS expression and NO production was observed after simultaneous pretreatment with PD98059 and SB203580 (P < 0. 05). Expression of iNOS mRNA in PRAMs and NO production in PRAMs culture supernatants were down-regulated by inhibition of NF-kappaB by PDTC (P <0. 05).

CONCLUSIONCellular signal molecules of ERK, p38 MAPK, and NF-kappaB all participate in the expression of iNOS and NO production in PRAMs induced by HMGB1.

Animals ; Cells, Cultured ; Extracellular Signal-Regulated MAP Kinases ; antagonists & inhibitors ; physiology ; Flavonoids ; pharmacology ; HMGB1 Protein ; pharmacology ; Imidazoles ; pharmacology ; Macrophages, Alveolar ; metabolism ; Male ; NF-kappa B ; antagonists & inhibitors ; physiology ; Nitric Oxide Synthase Type II ; biosynthesis ; Proline ; analogs & derivatives ; pharmacology ; Pyridines ; pharmacology ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Thiocarbamates ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; physiology

BACKGROUNDThe classic glycine receptor (GlyR) in the central nervous system is a ligand-gated membrane-spanning ion channel. Recent studies have provided evidence for the existence of GlyR in endothelial cells, renal proximal tubular cells and most leukocytes. In contrast, no evidence for GlyR in myocardial cells has been found so far. Our recent researches have showed that glycine could protect myocardial cells from the damage induced by lipopolysaccharide (LPS). Further studies suggest that myocardial cells could contain GlyR or binding site of glycine.

METHODSIn isolated rat heart damaged by LPS, the myocardial monophasic action potential (MAP), the heart rate (HR), the myocardial tension and the activities of lactate dehydrogenase (LDH) from the coronary effluent were determined. The concentration of intracellular free calcium ([Ca(2+)](i)) was measured in cardiomyocytes injured by LPS and by hypoxia/reoxygenation (H/R), which excludes the possibility that reduced calcium influx because of LPS neutralized by glycine. Immunohistochemistry was used to detect the GlyR in myocardial tissue. GlyR and its subunit in the purified cultured cardiomyocytes were identified by Western blotting.

RESULTSAlthough significant improvement in the MAP/MAPD(20), HR, and reduction in LDH release were observed in glycine + LPS hearts, myocardial tension did not recover. Further studies demonstrated that glycine could prevent rat mycordial cells from LPS and hypoxia/reoxygenation injury (no endotoxin) by attenuating calcium influx. Immunohistochemistry exhibited a positive green-fluorescence signaling along the cardiac muscle fibers. Western blotting shows that the purified cultured cardiomyocytes express GlyR beta subunit, but GlyR alpha1 subunit could not be detected.

CONCLUSIONSThe results suggest that glycine receptor is expressed in cardiomyocytes and participates in cytoprotection from LPS and hypoxia/reoxygenation injury. Glycine could directly activate GlyR on the cardiomyocytes and prevent calcium influx into the cardiomyocytes.

Animals ; Blood Pressure ; drug effects ; Blotting, Western ; Calcium ; metabolism ; Cytoprotection ; Glycine ; pharmacology ; Heart ; drug effects ; physiology ; Heart Rate ; drug effects ; Immunohistochemistry ; L-Lactate Dehydrogenase ; secretion ; Lipopolysaccharides ; toxicity ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, Glycine ; analysis ; physiology

OBJECTIVETo evaluate the improvement of Kangshuai Yizhi Formula I ( I, KYF I) on: the learning and memory dysfunction in mice, and on the mechanism of the hippocampal cholinergic system and the nervous system of monoamine which are closely related to learning and memory function.

METHODSMice: in the low-, middle-, and high-dose KYF I groups were given low-, middle-, and high-dose KYF, respectively, by gastrogavage for 35 successive days. Animals in the control group and the model group were treated with distilled water. The acute learning and memory dysfunction model was established by injection of scopolamine from day 31, and Morris water maze was used to assess the behavior performance of scopolamine-induced model mice for five days. The activities of acetylcholinesterase (AChE), choline acetyl transferase (ChaT) and the content of monoamine neurotransmitters in hippocampus were measured. The activity of monoamine oxidase (MAO) in hippocampus and serum was also detected.

RESULTS(1) Compared with the control group, the: mean escape latency was shortened, and the frequency across the platform and the staying time at the platform area on the 5th day were decreased in the model group by Morris water maze test. The activities of AChE and MAO were increased, and the ChaT activity and monoamine neurotransmitter content were decreased as well. (2) The escape latency for 4 days in the low-, middle-, and high-dose KYF I groups was significantly shortened than that in the model group, with the shortest latency in the high-dose KYF I group (P<0.05, P<0.01). The frequency across the platform was significantly increased and the staying time at the platform was significantly prolonged in the middle- and high-dose KYF I groups (P<0.05, P<0.01). (3) As compared with the model group, the activity of ChaT and the content of monoamine neurotransmitters in the hippocampus were significantly increased, and the activities of AchE and MAO were significantly decreased in the hippocampus in the high-dose KYF I group (P<0.01).

CONCLUSIONSHigh-dose KYF I can significantly improve the learning and memory dysfunction: induced by scopolamine in mice. Its mechanism may be related to improving the central cholinergic system and regulating the hippocampal monoamine neurotransmitters.

Acetylcholinesterase ; metabolism ; Animals ; Behavior, Animal ; drug effects ; Choline O-Acetyltransferase ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hippocampus ; drug effects ; enzymology ; pathology ; Learning ; drug effects ; Male ; Memory Disorders ; blood ; drug therapy ; enzymology ; physiopathology ; Mice ; Monoamine Oxidase ; blood ; Neurotransmitter Agents ; metabolism ; Reaction Time ; Scopolamine Hydrobromide ; toxicity ; Time Factors

BACKGROUNDThe association between vulnerability of plaque assessed with intravascular ultrasound (IVUS) and plasma levels of fibrinolytic biomarkers was determined in patients with acute coronary syndrome (ACS). However, few data are available on the relationship between the levels of tissue type plasminogen activator (t-PA) and virtual histological intravascular ultrasound (VH-IVUS) signs of plaque instability.

METHODSEighty-nine patients with ACS were enrolled in the study. Blood was collected to measure t-PA levels by liquid phase bead flow cytometry. Eighty-nine nonbifurcate lesions (identified by coronary angiography and ECG) were investigated using IVUS before catheterization. IVUS radiofrequency data obtained with a 20 MHz catheter were analyzed with IVUS virtual histological software. The areas of plaque and media were calculated and lesions were classified into two groups: VH-IVUS derived thin cap fibroatheroma (VH-TCFA) and non-VH-TCFA plaque.

RESULTSPlasma t-PA level in the patients with TCFA was significantly lower than that with non-TCFA ((1489+/-715) pg/ml vs (2163+/-1004) pg/ml). Decreased plasma levels of t-PA were associated with plaque vulnerability. Plasma levels of t-PA correlated negatively with plaque plus media and necrotic core in plaque in patients with ACS.

CONCLUSIONSt-PA is an independent risk factor and a powerful predictor of vulnerable plaques. Decreased levels of t-PA may reflect instability of atherosclerotic plaques and might therefore serve as noninvasive determinants of those at high risk for consequent adverse events.

Acute Coronary Syndrome ; blood ; pathology ; Aged ; Coronary Artery Disease ; pathology ; Coronary Vessels ; pathology ; Female ; Humans ; Male ; Middle Aged ; Tissue Plasminogen Activator ; blood ; Ultrasonography, Interventional

OBJECTIVETo investigate the effect of multi-plane Hyaluronic acid (HA) injection for rhinoplasty.

METHODSThe HA was injected below or above the periosteum at the nasal bone, above the perichondrium at the cartilage portion of nose, and between the great alar cartilage at the nasal tip. The HA volume was 1-1.5 ml, according to the nose form and aesthetic assessment. Over-injection was not permitted. Touch-up injection could be performed one week after the first injection if need.

RESULTSFrom Jan. 2010 to Jan. 2012, 60 cases underwent rhinoplasty with HA injection. The patients were followed up for 10-13 months with satisfactory result. The effect lasted about 9 months with the longest period as 12 months and the shortest period as 6 months.

CONCLUSIONSGood results can be achieved with multi-plane HA injection for rhinoplasty.

Adult ; Female ; Follow-Up Studies ; Humans ; Hyaluronic Acid ; administration & dosage ; therapeutic use ; Injections ; Male ; Rhinoplasty ; methods ; Treatment Outcome ; Young Adult

OBJECTIVETo detect over-expression of AChR-gamma mRNA in rhabdomyosarcoma tissues by duplex RT-PCR and discuss its potential in diagnosis of rhabdomyosarcoma.

METHODSDuplex RT-PCR was applied to the simultaneous detection of AChR-alpha and gamma subunit messenger RNA in 17 cases of rhabdomyosarcoma (9 ERMS, 6 ARMS, 2 PRMS). 20 cases of non-rhabdomyosarcomous small round cell tumors (6 poorly differentiated synovial sarcomas, 6 ES/PNET, 6 lymphomas, 2 neuroblastomas) and three normal muscle samples were also detected for AChR-alpha and gamma mRNA by the same method.

RESULTSAChR-alpha and AChR-gamma mRNA were expressed in all the cases of rhabdomyosarcoma. The rate of quantity in both transcripts was AChR-gamma/AChR-alpha >or= 1, but the rate for three normal muscle samples was < 1. Cases of non-rhabdomyosarcomous small round cell tumors were all negative for AChR-gamma.

CONCLUSIONAChR-gamma mRNA expression detected by molecular genetic methods is useful in diagnosis and differential diagnosis of rhabdomyosarcoma.

Diagnosis, Differential ; Humans ; Protein Subunits ; RNA, Messenger ; analysis ; Receptors, Cholinergic ; genetics ; Receptors, Nicotinic ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Rhabdomyosarcoma ; diagnosis

Breast Neoplasms ; pathology ; Cell Culture Techniques ; methods ; Cells, Cultured ; Female ; Humans ; Immunohistochemistry ; methods ; Quality Control

To evaluate the efficacy and safety of interferon-alpha-2b (IFN-α-2b) in polycythemia vera patients(PV patient) with or without post-polycythemic myelofibrosis (post-PV MF), 30 patients with mutated JAK2V617F were enrolled in this study, from which 29 patients were evaluable. The percentage of mutated JAK2V617F allele (V617F%) was evaluated by real-time polymerase chain reaction (RT-PCR) before and after treatment with IFN-α-2b. The correlation of V617F allele burden with the major clinical outcomes was studied. Adverse effects appeared in patients was observed. The results showed that the median follow-up was 24 (12 - 42) months for 29 evaluable patients. Complete hematologic response was achieved in 10%, 48%, 72% and 78% of patients after treatment for 6, 12, 24 and 36 months respectively. The detection of V617F allele burden revealed that the molecular remission of patients (V617F%) was achieved in 41%, 76%, 89% and 89% after treatment for 6, 12, 24 and 36 months respectively. Molecular complete remission (JAK2V617F undetectable) was achieved in 4 patients, lasted from 6 to 12 months after IFN-α-2b discontinuation. The decrease of V617F% in patients with post-PV MF was significantly higher than that in patients without post-PV MF (53 ± 18% vs 32 ± 22%, respectively; p = 0.031) after treatment for 12 months. PV patients had a good tolerance to IFN-α-2b. It is concluded that IFN-α-2b can decrease the mutated V617F allele burden. Patients with PV, especially with post-PV MF, can achieve molecular remission after treatment with IFN-α-2b.
Adult ; Alleles ; Female ; Humans ; Interferon-alpha ; therapeutic use ; Janus Kinase 2 ; genetics ; Male ; Middle Aged ; Mutation ; Polycythemia Vera ; drug therapy ; genetics ; pathology ; Primary Myelofibrosis ; drug therapy ; genetics ; pathology ; Recombinant Proteins ; therapeutic use

OBJECTIVETo study the circulating vaccine-derived poliovirus (cVDPVs) that occurred in Zhenfeng county, Guizhou province in 2004 and to discover wild-poliovirus, vaccine-derived poliovirus (VDPVs) and other vaccine-associated poliovirus which could cause clinical poliomyelitis.

METHODSField epidemiological studies at the epidemic area and collecting acute flaccid paralysis (AFP) case and contact stool specimen for virus identification and nucleotide sequencing. Analysis on data related to annual reports on stool specimens surveillance which involved AFP case and contacts in the resent years in Zhenfeng county.

RESULTSType-I VDPVs had been isolated from 2 AFP cases and 3 contact stool specimen in Wanlan village of Zhenfeng. After the first cVDPVs case was identified, there were 3 cases identified of having other vaccine-associated poliovirus of type-I or type-II in the 5 case of AFP that met the criteria of clinical poliomyelitis. The result of virological surveillance on polio showed that the EV isolation rate (55.1%) of Zhenfeng county was higher than the rate from the whole province of the same year (23.2%). The poliovirus (PV) isolation rate (36.8%) was obviously higher in 2004 than in the previous years. In the 16 PVs strains, the type-I accounted for 43.8% which was significantly higher than the average level (18.3%) from the whole province.

CONCLUSIONSData indicated that the type-I VDPVs had been circulating (cVDPVs) in Zhenfeng county in Guizhou province. Clinical poliomyelitis was caused by non-VDPVs. The increased PV infection and the decreasing rate of vaccination in the general population were responsible for the epidemic of type-I cVDPVs at this time. Monitoring and evaluation on the rate of routine immunization program and prediction of the trend of epidemic should be strenthened.

Adolescent ; Animals ; Cell Line ; Child, Preschool ; China ; epidemiology ; Disease Outbreaks ; Feces ; virology ; Humans ; Infant ; Infant, Newborn ; Male ; Paraplegia ; epidemiology ; etiology ; virology ; Poliomyelitis ; prevention & control ; Poliovirus ; immunology ; physiology ; Poliovirus Vaccines ; immunology ; Vaccination ; statistics & numerical data

OBJECTIVETo study the therapeutic effect of intravenous high-dose vitamin C on implanted hepatoma in rats.

METHODSThe rats bearing implanted Walker-256 hepatoma were treated with high-dose vitamin C at 2.83 and 5.65 g/kg intravenously, and the general condition, liver functions (A/G, ALT, AST, GGT), tumor volume, and tumor growth of the rats were evaluated.

RESULTSThe A/G of the rats treated with 2.83 g/kg vitamin C was significantly higher, but the ALT and GCT were significantly lower than those of the model rats (P<0.05 or 0.01). The ALT level in rats with 5.65 g/kg vitamin C treatment was significantly lower than that of the model rats (P<0.05). The tumor necrosis rate was significantly higher in rats with 2.83 g/kg vitamin C treatment than in the model rats (P<0.05).

CONCLUSIONIntravenous administration of 2.83 g/kg vitamin C can promote the necrosis and apoptosis of hepatoma Walker256 cells in rats and protect the liver function of the tumor-bearing rats.

Animals ; Apoptosis ; drug effects ; Ascorbic Acid ; administration & dosage ; Injections, Intravenous ; Liver Neoplasms, Experimental ; drug therapy ; pathology ; Male ; Necrosis ; Neoplasm Transplantation ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar