OBJECTIVETo study the effect of berberine on the release of nitric oxide (NO) by rat intestinal mucous microvascular endothelial cells (RIMECs) cultured in vitro for exploring the pharmacological mechanism of berberine in treating intestinal disease.

METHODSCultured RIMECs in vitro were identified adopting immunofluorescent stain with factor VIII-related antigen. NO level in the supernatant of normal cell culture or cell culture treated with different concentrations of berberine was detected with colorimetry at the 3rd, 6th, 9th and 12th h and compared.

RESULTSCompared with that in the normal control group, NO level increased more obviously in the berberine treated groups, and the best effect was shown in the 5 microg/mL berberine treated group.

CONCLUSIONOne of the important pharmacological mechanisms of berberine might be through promoting the endogenous NO release to induce endothelium-dependent dilatation of microvascular in intestinal mucosa, thus to improve the local microcirculation of intestine.

Animals ; Berberine ; pharmacology ; Cells, Cultured ; Colorimetry ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Intestinal Mucosa ; blood supply ; cytology ; Nitric Oxide ; biosynthesis ; Rats ; Time Factors

OBJECTIVETo investigate the properties of screw-bone interface of expansive pedicle screw (EPS) in osteoporotic sheep using micro-CT and histology.

METHODSSix female sheep with bilateral ovariectomy-induced osteoporosis were employed in this experiment and were randomly assigned into 2 groups: A and B. After EPS insertion in each femoral condyles, sheep in group A were bred for 3 months, while those in group B 6 months. Femoral condyles with EPS were 3D-imaged and reconstructed by micro-CT. Histology was evaluated thereafter.

RESULTSThe trabecular microstructure was denser at the screw-bone interface than in the distant parts in expansive section, especially within spiral marking. In the non-expansive section, however, there was no significant difference between the interface and the distant parts. The regions of interest (ROI) adjacent to EPS were reconstructed and analyzed by micro-CT using the same thresholds. The 3D-parameters generated, including tissue mineral density (TMD), bone volume fraction (BVF, BV/TV), bone surface/bone volume (BS/BV) ratio, trabecular thickness (Tb.Th), and trabecular separation (Tb.Sp), were significantly better in expansive than in non-expansive sections (P < 0.05). Histologically, newly formed bone trabeculae crawled along the expansive fissures and into the center of EPS. The newly formed bones, as well as the bone at the bone-screw interface, closely contacted the EPS and constructed four compartments.

CONCLUSIONSBased on micro-CT and histological evaluation, the study suggest that EPS can significantly provide stabilization in osteoporotic cancellous bone.

Animals ; Bone Density ; Bone Screws ; Disease Models, Animal ; Female ; Femur ; diagnostic imaging ; metabolism ; surgery ; Osteoporosis ; etiology ; pathology ; surgery ; Ovariectomy ; adverse effects ; Reproducibility of Results ; Sheep ; Tomography, X-Ray Computed ; methods

OBJECTIVETo investigate the properties of screw-bone interface of expansive pedicle screw (EPS) in osteoporotic sheep by micro-CT and histological observation.

METHODSSix female sheep with bilateral ovariectomy-induced osteoporosis were employed in this experiment. After EPS insertion in each femoral condyle, the sheep were randomly divided into two groups: 3 sheep were bred for 3 months (Group A), while the other 3 were bred for 6 months (Group B). After the animals being killed, the femoral condyles with EPS were obtained, which were three-dimensionally-imaged and reconstructed by micro-CT. Histological evaluation was made thereafter.

RESULTSThe trabecular microstructure was denser at the screw-bone interface than in the distant parts in expansive section, especially within the spiral marking. In the non-expansive section, however, there was no significant difference between the interface and the distant parts. The regions of interest (ROI) adjacent to EPS were reconstructed and analyzed by micro-CT with the same thresholds. The three-dimensional (3-D) parameters, including tissue mineral density (TMD), bone volume fraction (BVF, BV/TV), bone surface/bone volume (BS/BV) ratio, trabecular thickness (Tb.Th), and trabecular separation (Tb.Sp), were significantly better in expansive sections than non-expansive sections (P less than 0.05). Histologically, newly-formed bony trabeculae crawled along the expansive fissures and into the center of EPS. The newly-formed bones, as well as the bones at the bone-screw interface, closely contacted with the EPS and constructed four compartments.

CONCLUSIONSThe findings of the current study, based on micro-CT and histological evaluation, suggest that EPS can significantly provide stabilization in osteoporotic cancellous bones.

Animals ; Bone Screws ; Bone and Bones ; diagnostic imaging ; pathology ; Female ; Osteoporosis ; diagnostic imaging ; pathology ; Sheep ; Tomography, X-Ray Computed

OBJECTIVETo analyze the genetic differences of Orientia tsutsugamushi (Ot) Sta56 gene between Shandong isolates and other strains deposited in GenBank.

METHODSPCR and restriction fragment length polymorphism (RFLP) were used to amplify the complete sequence of Ot-Sta56 gene. RFLP profiles of Ot were predicted by a computer program according to their complete sequences of Ot-Sta56 gene. PCR amplicon from XDM2 strain was sequenced and analyzed by Clustal X (1.8) and PHYLIP software.

RESULTSThe complete sequences (about 1.6 kbp) of Ot-Sta56 gene were amplified from B16 strain (isolated from patients), FXS2 strain (isolated from A. agrarius) and XDM2 strain. Four species of restriction endonucleases (Hha I, Hinf I, Hae III, Pst I) were used to digest the PCR amplicons from the 3 isolates. When comparing with the RFLP profiles of prototype Ot, the RFLP profiles of PCR amplicons from the 3 isolates were similar to those of Japan Kawasaki strain, but were quite different from the international reference strains Gilliam, Karp, Kato. Results from DNA sequence analysis showed that the complete sequence of Ot-Sta56 gene homology to Japan Kawasaki strain of XDM2 strain was 97%, and deduced amino acid sequence was 92%.

CONCLUSIONData from the complete sequence of Sta56 gene indicated that the genotypes of Ot isolates in Shandong province were similar, but with distinction from the Kawasaki strain.

Amplified Fragment Length Polymorphism Analysis ; Antigens, Bacterial ; genetics ; Bacterial Proteins ; genetics ; Bacterial Typing Techniques ; classification ; DNA, Bacterial ; genetics ; Genes, Bacterial ; Membrane Proteins ; genetics ; Orientia tsutsugamushi ; genetics ; isolation & purification ; Polymorphism, Restriction Fragment Length ; Sequence Analysis, DNA

Aim To investigate the effect of chlorogenic acid (CGA) on chronic inflammatory pain induced by complete Freund's adjuvant (CFA) and to observe the influence of CGA on CFA-induced synaptic expression of AMAP receptor in spinal dorsal horn. Methods CFA was injected intraplantarly into the hindpaws of mice to induce mechanical allodynia and thermal hyperalgesia. The changes of paw withdrawal threshold (PWT) and the paw withdrawal latency (PWL) were tested after intrathecal administration of CGA. Meanwhile, the synaptic expression and phosphorylation of AMPA receptor subunit were assessed by immunoblot-ting. Results The intrathecal injection of CGA produced dose-depended improvement of both PWT and PWL in CFA injected mice. A higher dose of CGA (200ng) did not influence the base thresholds of normal mice. The median dose of CGA (100 ng) effectively reversed the CFA-induced synaptic expression of GluAl; meanwhile, it suppressed the phosphorylation of GluAl at Ser845, with no influence on phosphorylation at Ser831. Conclusions CGA might exert its analgesic effect by specifically inhibiting the phosphorylation of GluAl -Ser845 and the synaptic incorporation of GluAl-containing AMPA receptor.