OBJECTIVETo study the expression of the receptor for advanced glycation end products (RAGE) and the inhibitory effect of advanced glycation end products (AGEs) on testosterone production in rat Leydig cells.

METHODSRat Leydig cells were primarily cultured and the expression of RAGE in the Leydig cells was detected by RT-PCR and immunofluorescence staining. The Leydig cells were treated with AGEs at the concentrations of 25, 50, 100 and 200 microg/ml, respectively, and the testosterone content was determined by ELISA.

RESULTSRT-PCR and immunofluorescence staining exhibited the expression of RAGE in the rat Leydig cells. AGEs remarkably suppressed hCG-induced testosterone production in the Leydig cells in a concentration-dependent manner in the 50, 100 and 200 microg/ml groups as compared with the control (P < 0.01).

CONCLUSIONRAGE exists in rat Leydig cells and AGEs can significantly inhibit the secretion of testosterone in primarily cultured rat Leydig cells.

Animals ; Cells, Cultured ; Enzyme-Linked Immunosorbent Assay ; Glycation End Products, Advanced ; pharmacology ; Leydig Cells ; metabolism ; radiation effects ; Male ; Rats ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction ; Testosterone ; biosynthesis

Animals ; Cell Differentiation ; Cytokines ; metabolism ; Disease Progression ; Epithelial-Mesenchymal Transition ; Extracellular Matrix ; metabolism ; Fibroblasts ; metabolism ; pathology ; Hepatic Stellate Cells ; metabolism ; pathology ; Humans ; Liver ; metabolism ; pathology ; Liver Cirrhosis ; etiology ; metabolism ; pathology ; Stem Cells ; metabolism ; pathology

OBJECTIVE To have a systematic pathomechanism view of three chest impediment-syndromes of Qi Deficiency and Blood Stasis syndrome(QDBS),Qi Stagnation and Blood Stasis syn-drome (QSBS), Cold Obstruction and Qi Stagnation syndrome(COQS) and further investigate the changed metabolome and related pathways for screening potential biomarkers in rat plasma. METHODS According to clinical pathogeny, three kinds of syndrome models were established to simulate the disease of chest impediment. Plasma metabonomics based on UPLC-Q-TOF/MS was applied in this research to detected small molecule metabolites for identifyingthe special potential biomarkers of three chest impediment syndromes, respectively. RESULTS Significant metabolic differences were observed between thecontrol group and three syndrome groups. Furthermore, three syndrome groups were distinguished clearly by pattern recognition method.The particular metabolites contributing most to the classification of three chest impediment syndromes were identified. In the QSBS group, the potential biomarkers could include 2-keto-glutaramic acid, L-methionine, L-homocysteic acid, octadecanamide, stearoylglycine,behenic acid,linoleylcarnitine,lysoPC(14:1(9Z)),indoxyl sulfate and cholic acid.In the COQS group, they could be aminoadipic acid, palmitic amide, oleamide, lysoPC(P-16:0), lysoPC(P-18:0), lysoPC(20:2(11Z,14Z)), 9-HETE and tauroursodeoxycholic acid. Moreover, 4-pyridoxic acid, L-palmi-toylcarnitine, lysoPC(20:0), lysoPC (22:5 (4Z,7Z,10Z,13Z,16Z)), 3- hydroxyhexadecanoic acid and arachidonic acid could be the potential biomarkers for the QDBS group. CONCLUSION Three chest impediment syndromes have their own potential biomarkers.Each special metabolite has its owndifferent metabolic pathway.Both metabolismof cysteine and methionine,and metabolism of alanine,aspartate and glutamate are the main pathways in regulation of metabolic disorders in QSBS syndrome. Lysine biosynthesis and degradation,fatty acid metabolism,and glycerophospholipid metabolism are the main pathways in regulation of metabolic disorders in COQS syndrome.Arachidonic acid metabolism, fatty acid metabolism,fatty acid elongation in mitochondria,and vitamin B6 metabolism are the main pathways in regulation of metabolic disorders in QDBS syndrome.These endogenous substances were indicated as the special potential biomarkers for three chest impediment syndromes and worth studying in depth.

Objective To investigate rickettsiae infection from host animals and vector arthropods in some areas of Yunnan Province. Methods Rat clip and cage traps were used to capture mice. Chiggers from body surface of mice and ticks from body surface of farm cattle were collected. DNAs were extracted from mice spleens, chiggers and ticks. Rickettsiae groEL segment were amplified by nested-polymerase chain reaction (nPCR), sequenced to analyze the homology with other known sequences. Results A total of 410 samples were collected for rickettsiae groEL segment detection with nPCR and 19 samples (4.63%) showed positive for rickettsiae groEL segment . Among them, 2.68%(11/410)were positive for Orientia tsutsugamsushi (Ot) groEL segment, and 1.22%(5/410)were positive for spotted fever group rickettsia (SFGR) groEL segment, and 0.49%(2/410)were positive for rickettsia mooseri (Rm) groEL segment, and 0.24%(1/410)were positive for rickettsia endosymbiont(Re) groEL segment. When analyzed the homology with other known sequences, 11Ot strains with 93.6%-100% similarities among them in this study shared the highest similarity with other Ot strains from GenBank respectively, reached up to 96.1%-100%; The groEL segments of 5 SFGR strains with 92.1%-99.5% similarities among them in this study shared highest similarity with other SFGR strains from other GenBank respectively, reached up to 98.9%-100%; In this study groEL segments of 2 Rm strains all showed 100% similarity with Wilmington strain (GenBank No:AE017197); One groEL segment of Re showed 98.9% similarity with Re strain (GenBank No:EU435143). Conclusion There were kinds of rickettsiaes infection in host animals and vector arthropods in Yunnan Province, so the monitoring and prevention of the Rickettsiosis should be strengthened.

OBJECTIVETo investigate the effect of liquorice in functional modulation of intestinal P-glycoprotein (P-gp) in rats.

METHODSAn in vitro diffusion chamber system (Ussing chamber) was used to examine the direct effect of liquorice decoction on rhodamine 123 (a subtrate of P-gp) transport and evaluate the permeability of rhodamine 123 or fluorescein sodium through rat jejunum membranes after oral administration of liquorice decoction.

RESULTSDirect application of liquorice decoction did not obviously affect rhodamine 123 transport across the intestinal mucosa. Oral administration of liquorice decoction (10 g/kg, twice daily for a week) significantly increased the absorption of rhodamine 123 and also enhanced rhodamine 123 secretion across the jejunum mucosa. Liquorice had no obvious effect on the transport of CF across the jejunum mucosa.

CONCLUSIONLiquorice may slightly inhibit P-gp function in the intestinal mucosa to increase the intestinal absorption of rhodamine 123.

ATP-Binding Cassette, Sub-Family B, Member 1 ; antagonists & inhibitors ; metabolism ; Animals ; Glycyrrhiza ; Intestinal Absorption ; drug effects ; Intestinal Mucosa ; drug effects ; metabolism ; Intestines ; drug effects ; metabolism ; Male ; Plant Extracts ; administration & dosage ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Rhodamine 123 ; metabolism

Autoimmune Diseases ; diagnosis ; genetics ; therapy ; Humans ; Liver Diseases ; diagnosis ; genetics ; therapy

OBJECTIVETo evaluate the effect of cluster of differentiation 14 (CD-14) and Toll like receptors (TLR) on the expression of interleukin-6 (IL-6) mRNA induced by Porphyromonas endodontalis (Pe) lipopolysaccharides (LPS).

METHODSMC3T3-E1 cells were treated with 10 mg/L Pe-LPS for different hours, and the cells uninvolved by anything as the blank group. The expression of IL-6 was detected by reverse transcription polymerse chain reaction (RT-PCR) and enzyme-liked immunosorbent assay (ELISA). The expression of CD-14, TLR-2 and TLR-4 mRNA was observed at different time point (0 - 24 h) by RT-PCR. The protein of CD-14, TLR-2 and TLR-4 was analyzed with a flow cytometer. MC3T3-E1 cells were pretreated with anti-CD-14, anti-TLR-2 and anti-TLR-4 antibody for 1 h, and then cells were stimulated with 10 mg/L Pe-LPS for 6 h. The expression of IL-6 mRNA was examined by RT-PCR. Statistical analysis was performed using one-way ANOVA Dunnett-t test with SPSS 11.0 software package.

RESULTSThe IL-6 mRNA and proteins increased significantly after treatment with Pe-LPS. When MC3T3-E1 cells treated by Pe-LPS for 6 h, the expression of proteins soared from (11.696 ± 0.672) ng/L to (36.534 ± 0.574) ng/L (P < 0.01); In the control group, the CD-14 and TLR-4 mRNA are ambly-expression, and the ratios of CD-14 and TLR-4 positive cells were (39.038 ± 3.131)% and (11.438 ± 0.385)% respectively in MC3T3-E1. After treatment by Pe-LPS, the expression of CD-14 and TLR-4 mRNA increased significantly, and the ratios of CD-14 and TLR-4 positive cells markedly increased to (62.407 ± 1.800)% and (21.367 ± 2.271)%. TLR-2 expression did not change apparently after Pe-LPS treatment. The expression of IL-6 mRNA was partly inhibited by anti-CD-14 or anti-TLR-4 antibody, but not by TLR-2.

CONCLUSIONSPe-LPS can induce the expression of IL-6 in osteoblast MC3T3-E1 through CD-14 and TLR-4, but not TLR-2.

3T3 Cells ; Animals ; Antibodies ; immunology ; Interleukin-6 ; genetics ; metabolism ; Lipopolysaccharide Receptors ; genetics ; metabolism ; Lipopolysaccharides ; isolation & purification ; pharmacology ; Mice ; Porphyromonas endodontalis ; RNA, Messenger ; metabolism ; Toll-Like Receptor 2 ; genetics ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism

BACKGROUND:Periplaneta americana extract is recognized to have a positive effect on gastrointestinal mucosa. This study aimed to investigate the effects of periplaneta americana extract on immune function, nutrition status and gastrointestinal complications of early enteral nutrition patients with systemic inflammatory response syndrome (SIRS). METHODS:Patients with SIRS were randomly divided into two groups:treatment and control groups. All patients in the two groups received conventional therapy including enteral nutrition, but periplaneta americana extract, an additional Chinese medicine, was given to the patients in the treatment group. At the beginning of treatment (0 day) and 1, 3, and 7 days after treatment, the levels of immunoglobulin (IgA), total lymphocyte count (TLC), total protein (TP) and prealbumin (PA) were respectively tested in patients' venous blood. The incidences of bloating, diarrhea, aspiration pneumonia and high blood sugar at 7 days after treatment were recorded. The mortality of the patients in 28 days was recorded. RESULTS:At 3 and 7 days after treatment, the levels of IgA and TLC in the treatment group were higher than those in the control group (P<0.05). At 7 days after treatment, the levels of TP and PA in the treatment group were higher than those in the control group (P<0.05). The incidences of bloating and diarrhea in the treatment group were lower than those in the control group, the differences were significant (P<0.05). The mortality of treatment group was lower than that of the control group (P>0.05). CONCLUSION:Periplaneta americana extract could reduce gastrointestinal complications and improve immune function and nutritional status in patients with systemic inflammatory response syndrome.

Objective To evaluate the significance of platelet function monitoring in anti -platelet treatment of acute myocardial infarction . Methods The platelet maximum aggregation rates ( MAR ) induced by arachidonic acid , adenosine diphosphate and collagen were detected in 60 cases of acute myocardial infarction .The platelet aggregation function was compared before and after anti -platelet therapy for 24 h and 1 week.The plasma concentration of clopidogrel acid was detected and the correlation between concentration of clopidogrel acid and platelet aggrega-tion was analyzed.Results The MAR induced by arachidonic acid , adenosine diphosphate and collagen were significantly decreased after an-tiplatelet treatment for 24 h( P<0.01 );but after 1 week, no further re-duce in platelet aggregation rate was found , which showed some tole-rance.After 24 h, patients with MAR decreased by more than 30% ac-counted for 36.5%, 40.4% and 21.2%, respectively induced by arachidonic acid , adenosine diphosphate and collagen; patients with MAR decreased by 10% -29% accounted for 30.8%, 25%, and 34.6%,respectively,andthosewithMARdecreasedlessthan10%accountedfor34.6%,36.5%and44.2%,re-spectively.During half year follow -up, 34% of patients showed reoccurrence of cardiovascular events , and 55% of them were found aggregation rate less than 30%.Conclusion The aggregation rates under induction of arachidonic acid, adenosine diphosphate and collagen are all less than 30%when adopting the same anti -palate regime, but the incidence of cardiovascular events increased significantly and the platelet aggregation function and blood concentration of clopidogrel acid has no correlation .

Autophagy is a cellular degradation process for long-lived proteins and unnecessary or damaged organelles.The endoplasmic reticulum stress (ERS) response represents an adaptive mechanism that triggers the unfolded protein response to maintain ER homeostasis.Both ERS and autophagy were like double-edged sword,by which appropriate activity is cell protective,but excessive degree will trigger cell death.Several studies have suggested that ERS induced autophagy,which played the important role in myocardial ischemia reperfusion injury.This article reviews the recent advances about the effects and possible mechanism of ERS related autophagy on myocardial ischemia reperfusion injury.