Acute Developmental Toxicity of Panax notoginseng in Zebrafish Larvae / 中国结合医学杂志

OBJECTIVE@#To evaluate toxicity of raw extract of Panax notoginseng (rPN) and decocted extract of PN (dPN) by a toxicological assay using zebrafish larvae, and explore the mechanism by RNA sequencing assay.@*METHODS@#Zebrafish larvae was used to evaluate acute toxicity of PN in two forms: rPN and dPN. Three doses (0.5, 1.5, and 5.0 µ g/mL) of dPN were used to treat zebrafishes for evaluating the developmental toxicity. Behavior abnormalities, body weight, body length and number of vertebral roots were used as specific phenotypic endpoints. RNA sequencing (RNA-seq) assay was applied to clarify the mechanism of acute toxicity, followed by real time PCR (qPCR) for verification. High performance liquid chromatography analysis was performed to determine the chemoprofile of this herb.@*RESULTS@#The acute toxicity result showed that rPN exerted higher acute toxicity than dPN in inducing death of larval zebrafishes (P<0.01). After daily oral intake for 21 days, dPN at doses of 0.5, 1.5 and 5.0 µ g/mL decreased the body weight, body length, and vertebral number of larval zebrafishes, indicating developmental toxicity of dPN. No other adverse outcome was observed during the experimental period. RNA-seq data revealed 38 genes differentially expressed in dPN-treated zebrafishes, of which carboxypeptidase A1 (cpa1) and opioid growth factor receptor-like 2 (ogfrl2) were identified as functional genes in regulating body development of zebrafishes. qPCR data showed that dPN significantly down-regulated the mRNA expressions of cpa1 and ogfrl2 (both P<0.01), verifying cpa1 and ogfrl2 as target genes for dPN.@*CONCLUSION@#This report uncovers the developmental toxicity of dPN, suggesting potential risk of its clinical application in children.

Effect of Huangdi Anxiao Capsules on zebrafish vascular lesions induced by high glucose and high fat / 中国中药杂志

Zebrafish of different strains with 5 dpf (5 days post-fertilization) were selected and fed with 0.2% high-fat diet for 8 h and 3% glucose solution for 16 halternatively during the day and night for 4 consecutive days. The zebrafish model was established and randomly divided into model group, Huangdi Anxiao Capsules (260 mg·L⁻¹) group and pioglitazone (32 mg·L⁻¹) group. The drug treatment groups were given the water-soluble drugs, with a volume of 25 mL, and incubated in a 28 °C incubator for 4 days. To detect the exposure to the corresponding drugs, the normal control group was set up. Thirty zebrafish were included in each group. The effect of Huangdi Anxiao Capsules on vascular wall thickness, fluorescence intensity of islet beta cells, fluorescence intensity of macrophages, and blood flow velocity of zebrafish were detected. The expressions of vascular endothelial growth factor (vegfaa) and angiotensin converting enzyme (ACE) were detected by RT-PCR. The results showed that compared with the model group, Huangdi Anxiao Capsules can significantly reduce the thickness of the blood vessel wall, increase the fluorescence intensity of islet β cells and macrophages, increase the blood flow velocity in vivo, and decrease the ACE and vegfaa expressions in zebrafish. It is suggested that Huangdi Anxiao Capsules may alleviate zebrafish vascular lesions by regulating the expressions of ACE and vegfaa.

Application of multispectral animal living imaging technology in evaluating osteoarthritis model / 中国骨伤

<p><b>OBJECTIVE</b>To observe application value of multispectral animal living imaging technology in rats model of osteoarthritis.</p><p><b>METHODS</b>Fifteen male SD rats weighed (180 +/- 20) g (3 months old) were received intra-articular injection of iodoacetic acid for establishing osteoarthritis. Articular cavity of left knee of rats were injected into 50 microl iodoacetic acid. The same volume of sterile saline was injected into right knee articular cavity as control. X-ray living imaging and bone mineral density were observed at 2 and 4 weeks after establishment of model. After 4 weeks,rats were sacrificed and their bilateral joints were collected and determined histologically based on Collins classification and Kellgren-Lawrence classification.</p><p><b>RESULTS</b>Osteoarthritis model was successfully established, compared with control group, model group showed typical manifestation of osteoarthritis, including irregular cartilage surface,osteophyte formation,joint deformity and cartilage defect,and combined with significant decrease of bone density (P < 0.01), while the decrease was not obvious in proximal tibia (P < 0.05). After 2 weeks, knee joints in model group was classified as Collins grade 1 and Kellgren-Lawrence grade 2,then classified as Collins grade 4 and Kellgren-Lawrence grade 3 after 4 weeks,control group showed smooth articular surface,normal joint space and intact cartilage surface, knee joints was classified as Collins and Kellgren-Lawrence grade 0, and bone density of distal femur and proximal tibia were normal.</p><p><b>CONCLUSION</b>Multispectral animal living imaging technology could be used in dynamic observation of living imaging and detection of bone density in the animal model of osteoarthritis, and it is significant for evaluation of osteoarthritis model, and its realted tesearch.</p>

Different biological characteristics between nucleus pulposus and annulus fibrosus cells in rabbits / 中国骨伤

<p><b>OBJECTIVE</b>To compare biological characteristics between nucleus pulposus and annulus fibrosus cells in vitro model.</p><p><b>METHODS</b>Five New Zealand white rabbits (2 to 3 kg, either gender) were isolated nucleus pulposus and annulus fibrosus under sterilized condition, then cultured in nutrient solution with 15% FBS and DMEM/F12 (1:1) by enzyme digestion combined with tissue block method. When 90% cells fused, subcultring were performed. Cell morphology were observed by inverted phase contrast microscope, cell viability were detected by trypan blue staining, histological were observed by a toluidine blue and HE staining, cell proliferation were tested by MTT method, then the cell morphology, viability, proliferation between nucleus pulposus and annulus fibrosus were compared.</p><p><b>RESULTS</b>There were no obviously differences between nucleus pulposus and annulus fibrosus in original and the first strain. Physalides were appeared in annulus fibrosus on the second generation. The strapping time was later, and activity was lower in nucleus pulposus than annulus fibrosus. The growth of cell proliferation in nucleus pulposus was lower than annulus fibrosus from the ninth day.</p><p><b>CONCLUSION</b>The cell activity in annulus fibrosus is higher than nucleus pulposus. Digenerative disc disease may caused by recession of nucleus pulposus,local biomechnical changes, furether caused structure change and function loss of annulus fibrosus.</p>

Effects of Yougui recipe on the behavioral changes in rat of steroid-induced avascular necrosis of the femoral head / 中国骨伤

<p><b>OBJECTIVE</b>To explore the effect of Yougui Recipe, a kidney-supplementing and yang-activating formula which on the behavioral changes of rat of steroid-induced avascular necrosis of the femoral head (SANFH).</p><p><b>METHODS</b>Thirty Wistar rats were involved and were randomly divided into the blank control group (group A), the model group (group B) and the Yougui Recipe group (group C). SANFH models were established by injection of colibacillus endotoxin and prednisolone intramuscularly. Group C was lavaged with Yougui Recipe (10 ml/kg), while group A and group B were lavaged with the same amount of saline. The behavior of catch force, independent activities, the tail suspension, field experiment and water cleans maze experiment were observed after 6 weeks.</p><p><b>RESULTS</b>Compared with Yougui Recipe, rats in model group: catch force and independent activity decreases; the tail suspension activities was less time. In the desert field experiments, the total distance in 10 min movement reduced significantly. In the water maze experiment, incubation period of escape had a long time obviously, total distance of activities reduced.</p><p><b>CONCLUSION</b>Yougui Recipe can relieve the ethologic change of rat model of steroid-induced avascular necrosis of the femoral head.</p>

Research progress on proteomics in femur head necrosis / 中国骨伤

Appearance of proteomics technology can fleetly filt and reveal specificity biomarkers of disease, this will help to reveal the pathogenesis of femur head necrosis and help early diagnosis, find more effective methods and therapeutic targets. At present, they are hot spots that find out the occurred mechanism,related proteins of early diagnosis and early treatment and its functional identification; set up the early related database; optimize the protein extraction methods for research of femur head necrosis. This article reviews the application of study technology of related proteins of femur head necrosis on bone tissue, serum,related animal model,and in order to provide further research ideas.

Reseach of oxidative stress induces aging in rabbit intervertebral disc nucleus pulposus cells injured by H2O2 / 中国骨伤

<p><b>OBJECTIVE</b>To study the effect of H2O2 on the morphological pattern,vitality,proliferation,cycle period of rabbit intervertebral disc nucleus pulposus cells.</p><p><b>METHODS</b>Ten New Zealand white rabbits (2 to 3 kg, female) were used for isolating nucleus pulposus cells under sterilized condition. The culture solution with 15% FBS and DMEM/F12 (1:1) was applied for cell cultivation. After 90% cell fusion, the first generation was obtain and stimulated by H2O2 with different concentrations of 0 micromol/L (control group), 130 micromol/L,216 p.mol/L,360 Ipmol/L, 600 micromol/L,and 1000 micromol/L.</p><p><b>RESULTS</b>Compared with the control group, there was little difference of the biological property (P>0.05) in 130 micromol/L and 216 micromol/L H202-treated groups. When the concentration of H2O2 attained 360 micromol/L, 600 micromol/L, and 1 000 micromol/L, the cells suffered aging,with increased cell vacuoles,decreased proliferation,and aging:related increase of 13-galactosidase dyeing. The cell cycle of many nucleus pulposus cells was blocked in G1 stage other than entering S stage. With increasing H2O2 concentrations, the aging degree was increased.</p><p><b>CONCLUSION</b>A certain concentration of H202 could induce early aging of nucleus pulposus cells,resulting in biological abnormalities of these cells.</p>