Objective To determine mitochondria-associated signaling pathway on cell apoptosis in Leptospira interrogans infected murine mononuclear-macrophages. Methods A cell apoptotic model of L. interrogan serogroup Icterohaemorrhagiae strain Lai inducing apoptosis of murine mononuclear-macrophage line (J774A. 1) was established. Pathological changes of mitochondria in the infected cells were observed under transmission electron microscope. Mitochondrial potential and reactive oxygen species(ROS) levels in the infected cells were detected using JC-1 staining method and DCFH-DA fluorescent probe, respectively,and caspase-8/-9 activities in the infected cells were measured using commercial kits. and the apoptosis block effects of caspase inhibitors were determined by flow cytometry. Apoptosis in the infected cells and ap-optosis-blocking effect by caspase inhibitors were detected by flow cytometry. Western blot assay was adopted to examine the levels of cytochrome C (CytC) , AIF, EndoG and Smae in the mitochondria and cytoplasm.The transposition of either AIF or EndoG from the cytoplasm into cell nucleus was determined by immunoflu-orescence staining test. Results L. interrogans strain Lai could induce J774A. 1 cell apoptoais. In the in-fected cells, visible mitochondrial injury, declined mitochondrial membrane potential and elevated ROS level were presented. The activity of caspase-8 but not of caspase-9 was significantly increased. The caspase in-hihitors could not completely block the cell apoptosis. Both the AIF and EndoG was released from the mito-chondria and subsequently transferred from the cytoplasm into the nucleus in the infected cells. However,elevation of CytC level and Smac release in cytoplasm of the infected cells could not be found. Conclusion L.interrogans can induce apoptosis in the infected mononuclear-macrophage via AIF and EndoG belonging to caspase-independent pathway.

Rhei Rhizoma is a Chinese medicine with multiple botanical origins. There is a problem to identify it with conventional methods. To compare the characteristics of chloroplast matK gene sequences of different Rheum species and authenticate inspected species, the matK gene sequences of different species from different origins were amplified, cloned, and sequenced. Genomic DNA of Rheum plants was extracted using modified DNA extracted Kit and matK gene sequences were analyzed by ContingExpress, DNAman and MEGA5.0. The length of matK gene sequences of Rheum palmatum, R. tanguticum and R. officinale were 1 518 bp containing 57 variable loci. According to the mutation sites, R. palmatum, R. tanguticum and R. officinale were divided into different genotypes separately. Based on the established method according to the loci 587, 707, 838, we successfully identified the genuine Rheum species from its adulterants.

ObjectiveTo establish a method for content determination of total iridoid compounds and baldrinal and 11-ethoxyviburtinal from Valerianae Jatamansi Rhizoma et Radix; To determine the contents of total iridoid compounds and baldrinal and 11-ethoxyviburtinal in Valerianae Jatamansi Rhizoma et Radix from three medicinal origins.Methods UV spectrophotometry was applied, 11-ethoxyviburtinal (cyclopentane-pyran-7-formaldehyde, 4-ethoxy methyl) was set as the reference substance, and the content of total iridoid compounds was determined at 288 nm. HPLC method was used to simultaneously determine the contents of baldrinal and 11-ethoxyviburtinal. The HPLC analysis was performed on a Phenomenex Luna C18 column (250 mm×4.6 mm, 5μm). The mobile phase was composed of acetonitrile-water in gradient elution at a flow rate of 0.95 mL/min. The detection wavelength was 288 nm and the column temperature was 30℃.Results The total iridoid compounds, baldrinal and 11-ethoxyviburtinal were in good linearity within the ranges of 2.088–14.616μg/μL, 74.88–224.64μg, and 41.6–249.6μg, respectively. This method was precise, and with good repeatability, stability and recovery rate.Conclusion The method is accurate, simple, rapid, which can be used for the quality control of Valerianae Jatamansi Rhizoma et Radix.

Objective:To clarify the significance of the fractional anisotropy(FA) value and apparent diffusion coefficient(ADC) value of diffusion tensor imaging(DTI) applied in the glioma grading with Meta-analysis, and to provide a reference for selecting the treatment program and judging the prognosis of glioma.Methods:Computer search was performed in PubMed database, EMBASE database,CBM, CNKI, Wanfang database, and VIP database.The relevant neurosurgery magazines and neuroimaging magazines, the proceedings of academic meeting were retrieved by hand and the references of related reviews were retrospectively retrieved.Then the case-series studies according to inclusion and exclusion criteria were screened,and the qualities of included studies were evaluated.Meta-analysis was carried out by Review Manager 5.2 software.Results:Nine studies were included.The FA value of low-grade glioma(LGG) in tumor centre was lower than that of high-grade glioma(HGG) (χ2=11.94,df=8,P=0.15;I2=33%,Z=2.78,P=0.005;WMDFA=-0.02,95%CI:-0.03--0.01),and the ADC value of LGG in tumor centre was higher than that of HGG (χ2=5.26,df=4,P=0.26;I2=24%,Z=2.19,P=0.03;WMDADC=0.15,95%CI:0.02-0.28).Conclusion:When the FA value in tumor centre of the glioma is increasd,the ADC value is decreased,and the grade of glioma is increasd.The FA value and ADC value of DTI can help determine the grade of gliomas.

Objective To evaluate the feasibility and efficacy of intravascular optical coherence tomography (OCT) in the assessment of plaque characteristics and drug eluting stent deployment quality in the elderly patients with unstable angina (UA) and non-ST segment elevation myocardial infarction (NSTEMI). Methods OCT was used in elderly patients undergoing percutaneous coronary interventions.Fifteen patients, 9 males and 6 females with mean age of 72.6±5.3 years (range 67-92 years) were enrolled in the study. Images were obtained before initial balloon dilatation and following stent deployment. The plaque characteristics before dilation, vessel dissection,tissue prolapse, stent apposition and strut distribution after stent implantation were evaluated. Results Fifteen lesions were selected from 32 angiographic lesions as study lesions for OCT imaging after diagnostic coronary angiography. There were 7 lesions in the left anterior descending artery, 5 lesions in the right coronary artery and 3 lesions in the left circumflex coronary artery. Among them,12 (80.0%) were lipid-rich plaques, and 10 (66.7%) were vulnerable plaques with fibrous cap thickness 54.2±7.3 μm. Seven ruptured culprit plaques (46.7%) were found; 4 in UA patients and 3 in NSTEMI patients. Tissue prolapse was observed in 11 lesions (73.3%).Irregular stent strut distribution was detected in 8 lesions (53.3%). Vessel dissections were found in 5 lesions (33.3%). Incomplete stent apposition was observed in 3 stents (20%) with mean spacing between the struts and the vessel wall 172±96 mm (range 117-436 mm).Conclusions 1) It is safe and feasible to perform intravascular OCT to differentiate vulnerable coronary plaque and monitor stent deployment in elderly patients with UA and USTEMI. 2) Coronary plaques in elderly patients with UA and USTEMI could be divided into acute ruptured plaque, vulnerable plaque, lipid-rich plaque, and stable plaque. 3) Minor or critical plaque rupture is one of the mechanisms of UA in elderly patients. 4) Present drug eluting stent implantation is complicated with multiple tissue prolapses which are associated with irregular strut distributions. 5) The action and significance of tissue prolapse on acute vessel flow and in-stent thrombus and restenosis need to be further studied.

Using recombinant TpNs proteins of Treponema pallidum as antigens, ELISAs are proved to be of higher sensitivity and specificity. However, they can be further increased by using multiple TpNs antigens. According to the epitope analysis, we firstly used linking primers PCRs to obtain an artificial fusion gene segment tpE17-47 containing epitopes of both TpN17 and TpN47. Subsequently, we conducted the prokaryotic expression systems of entire tpN17 and tpN47 genes and tpE17-47 fusion gene. SDS-PAGE analysis and BioRad Gel Image Analysis System showed that the recombinant proteins rTpN17, rTpN47 and rTpE17-47 expressed stably, with 36%, 20% and 28% yields of total bacterial protein, respectively. After purified by Ni-NTA affinity chromatography, all the three recombinant proteins could be recognized by T. pallidum antibody positive sera from syphilis patients. The positive rate of rTpE17-47-ELISA for detecting serum specimens in clinically 630 cases with syphilis was 98.6%. This rate was slightly higher than that by Treponema pallidum particle agglutination (TPPA) (97.9%) (P > 0.05), but significantly higher than those by rTpN17-ELISA (83.8%), rTpN47-ELISA (83.3%) and rapid plasma reagin (RPR) (72.1%) (P < 0.01). Furthermore, both ELISAs and TPPA for detecting the serum specimens in 25 cases with SLE, 36 cases with RA and 250 healthy cases were all negative. RPR showed positive in 1 case with SLE, 2 cases with RA and 2 healthy cases. This could be a novel serological screening or diagnostic method of syphilis with advantages of quickness, convenience, safety, sensitivity and specificity.
Amino Acid Sequence ; Antigens, Bacterial ; biosynthesis ; genetics ; Base Sequence ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunodominant Epitopes ; immunology ; Molecular Sequence Data ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Sensitivity and Specificity ; Syphilis ; diagnosis ; microbiology ; Syphilis Serodiagnosis ; methods ; Treponema pallidum ; immunology

OBJECTIVETo investigate the changes in aorta morphology and Ca(2+)-activated K(+) (KCa) channel expression in the diabetic rats.

METHODSA diabetic rat model was established by a single intraperitoneal injection of streptozotocin (30 mg/kg) after a modified high fat and glucose diet for 8 weeks. Pathological changes in the aorta were observed with HE staining, elastic fiber staining, Masson's trichrome staining and immunohistochemistry. Both the mRNA and protein levels of KCa channels in the aorta were measured by RT-PCR and Western blotting.

RESULTSEarly atherosclerotic changes were observed in the aorta wall of the diabetic rats. The mRNA and protein levels of KCa1.1 channel α- and β-subunits were significantly decreased, while the expression of KCa3.1 channels was obviously enhanced in the middle layer of the aorta in the diabetic rats.

CONCLUSIONKCa channel switching in smooth muscles may play a role in the development of atherosclerosis in diabetic rats.

Animals ; Aorta ; pathology ; Atherosclerosis ; pathology ; Diabetes Mellitus, Experimental ; pathology ; Intermediate-Conductance Calcium-Activated Potassium Channels ; metabolism ; Large-Conductance Calcium-Activated Potassium Channel alpha Subunits ; metabolism ; Male ; Muscle, Smooth, Vascular ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To investigate the effects of 11-ethoxyviburtinal of Valeriana jatamansi Jones on the expression of enterochromaffin cells ( EC) and mast cells ( MC) in rats with irritable bowel syndrome ( IBS) . Methods The IBS rat model was induced by chronic unpredictable stress with isolation. Male SD rats were randomly divided into six groups:control group,model group, pinaverium group at the dose of 25 mg/kg,and high-, mid-and low-dose of 11-ethoxyviburtinal group at dose of 1. 2, 0. 6, and 0. 3 mg/kg respectively. The number of 5-HT positive immunoreactive colonic EC was detected by the method of immunohistochemistry and that of colonic MC was measured with toluidine blue staining. Results Compared with control group, the number of 5-HT positive immunoreactive colonic EC and MC increased significantly in model group ( P <0 . 05 ) . The high dose of 11-ethoxyviburtinal could significantly de-crease the number of colonic EC and MC ( P<0 . 05 ) . Conclusion The 11-ethoxyviburtinal of Valeri-ana jatamansi Jones could significantly reduce the number of 5-HT positive immunoreactive 5-HT positive immunoreactive EC and MC in colon tissue of rats with IBS.

OBJECTIVETo investigate the mechanism of iridoid from Valeriana jatamansi treating irritable bowel syndrome.

METHODSixty male SD rats were equally divided into 6 groups (2 controls, 1 model and 3 treatment doses) with 10 rats per group. The test groups were administered with iridoid (24.92, 12.46, 6. 23 mg x kg(-1)) while the control groups were administered with fluoxetine (2.5 mg x kg(-1), positive control) or distilled water (negative control). The model was established by chronic stress and independent feeding. The influence of iridoid from V. jatamansi on 5-HT and 5-HIAA in colon, serum and hypothalamic were observed in all groups.

RESULTIn the model group, the content of 5-HT in colon and serum increased significantly, but the content of 5-HT in hypothalamic decreased significantly. The content of 5-HIAA and the value of 5-HT/5-HIAA had no significant change. In three iridoid-treated groups, the content of 5-HT in colon and serum decreased, but the content of 5-HT in hypothalamic increased. The content of 5-HIAA had no significant change. The value of 5-HT/5-HIAA in colon and serum reduced.

CONCLUSIONThe mechanism of iridoid from V. jatamansi treating irritable bowel syndrome may be related to the regulation effect to the levels of 5-HT from Gastrointestinal to central nervous system.

Animals ; Chromatography, High Pressure Liquid ; Colon ; drug effects ; metabolism ; Hydroxyindoleacetic Acid ; blood ; metabolism ; Hypothalamus ; drug effects ; metabolism ; Iridoids ; therapeutic use ; Irritable Bowel Syndrome ; blood ; drug therapy ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Serotonin ; blood ; metabolism ; Valerian ; chemistry