OBJECTIVETo examine the role of lactoferrin (LF) on Toll like receptor 4 (TLR4) stimulated by lipopolysaccharide (LPS) in human periodontal ligament cells (hPDLCs).

METHODSPrimary hPDLCs were cultured by tissue block enzymolytic method. Cells obtained from four passages were identified and used in this experiment. Cells without stimulation served as the controls and cells treated with LPS (0.1 microg x mL(-1)) comprised the LPS group. The LPS + LF group was pretreated with LPS (0.1 microg x mL(-1)) for 2 h, and then treated with LF (10 microg x mL(-1)). Four hours after LF stimulation, the mRNA expression levels of TLR4 were examined by real-time quantitative polymerase chain reaction (RT-PCR). The protein expression of TLR4 was observed by cell immunofluorescence staining after LF stimulation of 24 hours.

RESULTSTLR4 mRNA expression in the LPS + LF group was significantly more decreased than that in the LPS group (P < 0.05), but exhibited no difference with that in the control group (P > 0.05). Cell immunofluorescence staining showed that the protein expression of TLR4 in the LPS + LF group was significantly more decreased than that in the LPS group (P < 0.05), but exhibited no difference with that in the control group (P > 0.05).

CONCLUSIONLF can decrease the expression of TLR4 stimulated by LPS in hPDLCs, thus presenting potential application for controlling the TLR4 immune pathway of periodontitis.

Down-Regulation ; Humans ; Lactoferrin ; Lipopolysaccharides ; Periodontal Ligament ; Periodontitis ; Toll-Like Receptor 4

Ginsenosides are the abundant secondary metabolites in American ginseng (Panax quinquefolium), it could be released into soil through root exudation and decomposition during plant growth. This study determined ginsenoside contents in American ginseng cultivated soil by HPLC. Three ginsenosides, Rb1, Rb2 and Rd, were detected in the rhizosphere soil of 3-4 years old American ginseng cultivated in Huairou District, Beijing, and their contents were 0.80-3.19 mg x kg(-1). Correspondingly, the contents of the three ginsenosides in soil solution were 4-16 mg x L(-1) at field water-holding capacity of 20%. According to the field soil test data, we designed the concentration of ginsenosides for bioassays (0.2-125 mg x L(-1) in solution or 0.2-125 mg x kg(-1) in soil). The results showed that radicle lengths of American ginseng were reduced by 6%-23% in solution containing 0.2-125 mg x L(-1) ginsenoside extract, and a significant difference was observed at concentration of 125 mg x L(-1) (P < 0.05). The shoot lengths of American ginseng were not significantly inhibited by 0.2-125 mg x L(-1) ginsenosides extractions. After 20 days of growth in nutrient solution amended with 25 mg x L(-1) ginsenosides extraction, plant height of 3-year-old American ginseng seedling was decreased by 28% compared to the control, and the biomass of aerial parts was also reduced by 50% (P < 0.05). However, the growth of newly-grown fibrous root was not significantly inhibited. Comparatively, when American ginseng embryos were cultivated into sterile or non-sterile soil, neither radicle lengths nor shoot lengths were significantly affected by 0.2-125 mg x kg(-1) ginsenoside extracts. In conclusion, ginsenosides showed autotoxic effect on growth of American ginseng radicle and adult seedling, however, this effect was weakened in field soil.
Chromatography, High Pressure Liquid ; Culture Media ; chemistry ; metabolism ; Ginsenosides ; analysis ; metabolism ; toxicity ; Panax ; chemistry ; drug effects ; growth & development ; metabolism ; Plant Roots ; chemistry ; drug effects ; growth & development ; metabolism ; Soil ; chemistry

OBJECTIVETo determine the potential urinary biomarkers of metabolic syndrome (MS) with early renal injury and establish diagnostic models by magnetic bead-based separation and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS).

METHODSParticipants were selected from the epidemiologic study on MS and renal involvement among residents in Pinggu district, Beijing. Eight-hour overnight urine samples were fractionated by means of magnetic bead-based weak cation exchange chromatography and subsequently analyzed with MALDI-TOF-MS. Wilcoxon test and random forests were used to screen differential protein peaks of MS patients with early renal injury, then combined with genetic algorithm and support vector machine, respectively, to establish diagnostic models.

RESULTSTotally 54 cases of MS without renal injury and 46 cases of MS with early renal injury were enrolled. Totally twenty protein peaks were up-regulated in the urine of MS patients with early renal injury by Wilcoxon test (P < 0.05); random forests algorithm revealed twelve protein peaks up-regulated in the urine of MS patients with early renal injury (importance value of mean decrease in accuracy > 0.005). Genetic algorithm based model showed 82.6% sensitivity, 84.3% specificity, and 83.5% accuracy by a 10-fold cross-validation in identifying MS patients with early renal injury; correspondingly, the support vector machine based model reported 89.2% sensitivity, 81.1% specificity and 85.5% accuracy. Four protein peaks were included in two diagnostic models with mass-to-charge ratios of 2756.98, 3019.11, 9077.04, and 10 054.26.

CONCLUSIONSThe urinary proteome patterns of MS with early renal injury were successfully established with magnetic bead-based separation and MALDI-TOF-MS technology. A series of urinary differential expressing protein peaks were identified with bioinformatics tools. Diagnostic models combining cluster of protein peaks are capable of differentiating MS patients with early renal injury from those without renal injury. The different urine protein excretion patterns revealed in this study provide urinary candidate biomarkers of MS patients with early renal injury for future identification and biological roles investigation.

Adult ; Biomarkers ; urine ; Chromatography, Ion Exchange ; methods ; Female ; Humans ; Kidney Diseases ; etiology ; urine ; Male ; Metabolic Syndrome ; complications ; urine ; Middle Aged ; Proteome ; analysis ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods ; Urine ; chemistry

BACKGROUND: High-volume hemofiltration (HVHF) is technically possible in severe acute pancreatitis (SAP) patients complicated with multiple organ dysfunction syndrome (MODS). Continuous HVHF is expected to become a beneficial adjunct therapy for SAP complicated with MODS. In this study, we aimed to explore the effects of fluid resuscitation and HVHF on alveolar-arterial oxygen exchange, the Acute Physiology and Chronic Health Evaluation II (APACHE II) score in patients with refractory septic shock. METHODS: A total of 89 refractory septic shock patients, who were admitted to ICU, the Provincial Hospital affiliated to Shandong University from August 2006 to December 2009, were enrolled in this retrospective study. The patients were randomly divided into two groups: fluid resuscitation (group A, n=41), and fluid resuscitation plus high-volume hemofiltration (group B, n=48). The levels of O2 content of central venous blood (CcvO2), arterial oxygen content (CaO2), alveolar-arterial oxygen pressure difference P(A-a)DO2, ratio of arterial oxygen pressure/alveolar oxygen pressure (PaO2/PAO2), respiratory index (RI) and oxygenation index (OI) were determined. The oxygen exchange levels of the two groups were examined based on the arterial blood gas analysis at different times (0, 24, 72 hours and 7 days of treatment) in the two groups. The APACHE II score was calculated before and after 7-day treatment in the two groups. RESULTS: The levels of CcvO2, CaO2 on day 7 in group A were significantly lower than those in group B (CcvO2: 0.60±0.24 vs. 0.72±0.28, P<0.05; CaO2: 0.84±0.43 vs. 0.94±0.46, P<0.05). The level of oxygen extraction rate (O2ER) in group A on the 7th day was significantly higher than that in group B ( 28.7±2.4 vs. 21.7±3.4, P<0.01). The levels of P(A-a)DO2 and RI in group B on the 7th day were significantly lower than those in group A. The levels of PaO2/PAO2 and OI in group B on 7th day were significantly higher than those in group A (P<0.05 or P<0.01). The APACHE II score in the two groups reduced gradually after 7-day treatment, and the APACHE II score on the 7th day in group B was significantly lower than that in group A (8.2±3.8 vs. 17.2±6.8, P<0.01). CONCLUSION: HVHF combined with fluid resuscitation can improve alveolar- arterial-oxygen exchange, decrease the APACHE II score in patients with refractory septic shock, and thus it increases the survival rate of patients.

OBJECTIVETo evaluate the complications of CO2 laser surgery in the treatment of laryngeal carcinoma, to analyze related factors and to propose preventive measures.

METHODSRetrospective analysis of 912 cases of laryngeal carcinoma (35 cases of supraglottic cancer and 877 cases of glottic cancer) treated only with laser surgery in Tongren Hospital was carried out. Among the glottic cancer, carcinoma in situ (Tis), T1, T2 and T3 were 53, 659, 158 and 7 cases. The follow-up period ranged from 2 to 18 years, with a median follow-up time of 9.3 years.

RESULTSOf 912 cases, 824 cases were still alive, 29 cases failed to be followed-up (taken into dead number), and 59 cases were dead. The recurrent rate was 9.4% (86/912). Three year survival rate was 95.6% (775/811) and five year survival rate was 87.9% (518/589). The incidence of surgery complications was 9.1% (83/912). Incidence of complications in supraglottic carcinoma and glottic carcinoma were 17.1%(6/35) and 8.8% (77/877), respectively, with no difference between the two groups (χ(2) = 2.85, P > 0.05). Incidence of complications of Tis, T1, T2 and T3 cases of glottic cancer were 5.7%(3/53), 7.8% (51/659), 13.3% (21/158) and 28.6% (2/7) respectively, with significant difference (χ(2) = 8.97, P < 0.05). Incidence of complications of glottic carcinoma with and without anterior commissure incision were 12.8%(31/242) and 7.2%(46/635) respectively, with significant difference between the two groups (χ(2) = 6.78, P < 0.05). Incidence of complications in the patients underwent type II, III, IV, V cordectomy were 3.8% (4/105), 7.0% (20/287), 9.7% (22/226) and 12.0% (31/259) respectively, with significant difference (χ(2) = 7.96, P < 0.05).

CONCLUSIONSThere are some potential risks and complications intra- and post-operatively, according to the sites and extent of the primary tumors and the range and depth of removed tissues. It needs to take active preventive measures to reduce the incidence of complications.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Intraoperative Complications ; etiology ; prevention & control ; Laryngeal Neoplasms ; surgery ; Laser Therapy ; adverse effects ; Lasers, Gas ; adverse effects ; Male ; Middle Aged ; Postoperative Complications ; etiology ; prevention & control ; Retrospective Studies

OBJECTIVETo analyze the effects of continuous cropping and soil treatment on rhizosphere fungal community of Panax quinquefolium, and the correlation between rhizosphere fungal community and growth of P. quinquefolium.

METHODField plot trail of continuous cultivated P. quinquefolium was conducted, meanwhile continuous cropping soil was treated by organic carbon fertilizer plus methylene dithiocyanate and Kingbo respectively. The rate of seedling survival, root disease index and root weight were investigated. Dilution plating was used to analyze the rhizosphere fungal community, multiple comparisons including H, M(a) and J were performed between rhizosphere fungal community and growth of P. quinquefolium.

RESULTAs to P. quinquefolium planted in continuous soil, the rate of seedling survival, root weight decreased, root disease index increased remarkably (P<0.05). Meanwhile the fungal counts decreased 50% -63% , the diversity index (H') decreased 39%-43%, while the ratio of Penicillium and Aspergillus increased. There was a significant negative correlation between rhizosphere fungal diversity and P. quinquefolium root disease index (r = -0.970, P=0.006). Organic carbon fertilizer plus methylene dithiocyanate treatment could improve the rate of seedling survival and root weight, and could partly increase the rhizosphere fungal diversity.

CONCLUSIONIn continuous cropping soil of P. quinquefolium, the rhizosphere fungal counts and diversity index decreased, meanwhile the fungal community composition changed. Organic carbon fertilizer plus methylene dithiocyanate treatment could improve the growth of continuous cultivated P. quinquefolium.

Agriculture ; methods ; Biodiversity ; Fertilizers ; analysis ; Fungi ; classification ; drug effects ; growth & development ; isolation & purification ; Panax ; growth & development ; microbiology ; Plant Diseases ; microbiology ; Plant Roots ; growth & development ; microbiology ; Rhizosphere ; Soil Microbiology

OBJECTIVETo investigate the influence of intranasal instilling titanium dioxide (TiO2) nanoparticles on monoaminergic neurotransmitters at different-time exposure.

METHODSCD female mice were intranasally instilled three different-sized (25 nm, 80 nm and 155 nm) TiO, suspension every other day in a dose of 50 mg/kg body weight. The control group was instilled the same volume of Milli-Q water. Inductively coupled plasma-mass spectrometry (ICP-MS) was used to analyze the titanium contents in murine brain after exposure to TiO2 particles 2 days, 10 days, 20 days and 30 days. The monoaminergic neurotransmitters such as norepinephrine (NE), dopamine (DA), 5-hydroxytryptamine (5-HT), 5-hydroxyindole acetic acid (5-HIAA), 3, 4-dihydroxyphenylacetic acid (DOPAC), and homovanillic (HVA), were determined by reversed-phase high performance liquid chromatography (RP-HPLC) with electrochemical detector.

RESULTSAfter exposure to TiO, nanoparticles 10 days, the titanium contents in murine brain were increased, the titanium content in the 25 nm group was up to (1059.3 +/- 293.5) ng/g. In 20 days, the titanium content decreased slowly with the metabolism of titanium in vivo, but it kept at a high level, the content decreased to (654.7 +/- 269.2) ng/g in the 25 nm group. After exposure to TiO2 nanoparticles 30 days, the titanium contents had no obviously change. Because of the accumulation of TiO, in the brain, the contents of NE and 5-HT increased significantly after exposure to 80 nm and 155 nm TiO, nanoparticles 20 days, while the decreased contents of DA, DOPAC, HVA and 5-HIAA were observed.

CONCLUSIONThe inhaled TiO2 nanoparticles could be translocated to and deposited in murine brain after absorbing by nasal mucosa, and further influence the releases and metabolisms of monoaminergic neurotransmitters in brain.

Administration, Intranasal ; Animals ; Biogenic Monoamines ; metabolism ; Brain ; drug effects ; metabolism ; Brain Chemistry ; Female ; Metal Nanoparticles ; Mice ; Neurotransmitter Agents ; metabolism ; Time ; Titanium ; administration & dosage ; pharmacology

AIMTo investigate the effects of ramipril on myocardial ischemia/reperfusion injury in diabetic rats, and to explore its mechanism according to the observation on myocardial ultrastructure.

METHODSStreptozotocin induced diabetic rats were divided randomly into three groups (n = 16): ischemia/reperfusion (I/R), ischemic preconditioning (IPC) and ramipril (RAM) group. Rats in RAM group were administered by RAM(1 mg x kg(-1) x d(-1)) orally for 4 weeks, the others were administered by normal saline. Then all rats were subjected to myocardial ischemia/ reperfusion injury. Rats in IPC group were preconditioned before ischemia. The ECG and the infarct size were examined. The changes of myocardial morphology were examined by light and electron microscopes.

RESULTSCompared with I/R group, the elevation of ST segment and the incidence of ventricular tachycardia and ventricular fibrillation during ischemia were significantly decreased, the infarct size at the end of reperfusion was remarkably reduced, the myocardial morphology were significantly improved, special structure of myofilaments and mitochondria remained clearly, blood vessels were unobstructed, injury of endothelium were decreased in PC and RAM groups.

CONCLUSIONRamipril administered for 4 weeks induces myocardial protection in diabetic rats, which is similar to that of IPC. The mechanism may be involved in protection of cardiocytes and mitochondria, and improvement of endothelial function.

Animals ; Cardiotonic Agents ; pharmacology ; Diabetes Mellitus, Experimental ; complications ; Ischemic Preconditioning, Myocardial ; methods ; Myocardial Reperfusion Injury ; pathology ; prevention & control ; Myocardium ; ultrastructure ; Ramipril ; pharmacology ; Rats

OBJECTIVETo search for the optimal approach for hepatocyte-directed differentiation of hepatic progenitor cells and investigate the molecular mechanism of the hepatic differentiation.

METHODSHepatic progenitor cells were infected with recombinant adenovirus which containing human LIF, BMP2 or BMP9 gene. The maturation and differentiation of progenitor cells were examined by PAS staining and ICG uptake methods at 4, 7 and 10 days post infection. The production of Albumin (Alb) was measured by luciferase activity at day 4, 7, 10 and 14.

RESULTSPAS staining assay revealed that BMP2 and BMP9 enhanced glycogen storage in hepatic progenitor cells most obviously at day 7. The percentages of positive cells were 30% and 45% respectively at 7 days post-infection. Meanwhile, 40% and 30% cells were positive by ICG uptake assay after BMP2 and BMP9 induction. Luciferase activity indicated that BMP9 induced ALB-Luc activity most significantly at day 7. However, less inductive activity was found in LIF-treated group.

CONCLUSIONThese results indicated tuat hepatic progenitor cells were differentiated into hepatocyte-like cells by BMPs and LIF induction.

Adenoviridae ; Bone Morphogenetic Proteins ; pharmacology ; Cell Differentiation ; Cells, Cultured ; Hepatocytes ; cytology ; metabolism ; virology ; Humans ; Leukemia Inhibitory Factor ; pharmacology ; Stem Cells ; cytology ; metabolism ; virology

OBJECTIVEMany children were found to have low free carnitine level in blood by tandem mass spectrometry technology. In some of the cases the problems occurred secondary to malnutrition, organic acidemia and other fatty acid oxidation metabolic diseases, and some of cases had primary carnitine deficiency (PCD). In the present article, we discuss the diagnosis of PCD and evaluate the efficacy of carnitine in the treatment of PCD.

METHODWe measured the free carnitine (C0) and acylcarnitine levels in the blood of 270 000 neonates from newborns screening program and 12 000 children with suspected clinical inherited metabolic diseases by tandem mass spectrometry. The mutations of carnitine transporter protein were tested to the children with low C0 level and the diagnosis was made. The children with PCD were treated with 100 - 300 mg/kg of carnitine.

RESULTSeventeen children were diagnosed with PCD, 6 from newborn screening program and 11 from clinical patients. Mutations were found in all of them. The average C0 level [(2.9 ± 2.0) µmol/L] in patients was lower than the reference value (10 µmol/L), along with decreased level of different acylcarnitines. The clinical manifestations were diverse. For the 6 patients from newborn screening, 4 were asymptomatic, 1 showed hypoglycaemia and 1 showed movement intolerance from 2 years of age. For the 11 clinical patients, 8 showed hepatomegaly, 7 showed myasthenia, 6 showed cardiomyopathy, 1 showed chronic abdominal pain, and 1 showed restlessness and learning difficulty. Among these patients, 14 cases were treated with carnitine. Their clinical symptoms disappeared 1 to 3 months later. The C0 level in the blood rose to normal, with the average from (4.0 ± 2.7) µmol/L to (20.6 ± 8.3) µmol/L (P < 0.01). However, the level was still lower than the average level of healthy children [(27.1 ± 4.5) µmol/L, P < 0.01].

CONCLUSIONSeventeen patients were diagnosed with PCD by the test levels of free carnitine and acylcarnitines in blood with tandem mass spectrometry, and gene mutation test. Large dose of carnitine had a good effect in treatment of the PCD patients.

Cardiomyopathies ; diagnosis ; drug therapy ; genetics ; Carnitine ; analogs & derivatives ; blood ; deficiency ; genetics ; Child, Preschool ; DNA Mutational Analysis ; Female ; Follow-Up Studies ; Humans ; Hyperammonemia ; diagnosis ; drug therapy ; genetics ; Infant ; Infant, Newborn ; Male ; Muscular Diseases ; diagnosis ; drug therapy ; genetics ; Mutation ; Neonatal Screening ; methods ; Organic Cation Transport Proteins ; deficiency ; genetics ; Reference Values ; Tandem Mass Spectrometry