OBJECTIVE:To establish the method to identify the polysaccharide element in Holothuria atra Jeager by spectroscopy.METHODS:The purity of polysaccharide in Holothuria atra Jeager was determined by electrophoretic method;the infrared absorption spectrum,ultraviolet absorption spectrum,carbon spectra and hydrogen spectra of NMR were identified in structure by infrared spectrophotometry,ultraviolet spectrophotometry and NMR spectrometry respectively.RESULTS:The gel band after electrophoresis took blue color and the results of spectrum determination accountable for each other.CONCLUSION:Uniformly constituent of polysaccharose was found in the Holothuria atra Jeager,which was an acidic polysaccharose with vitriolic esterfunction,without glucopeptide;?-configuration,?-glucopyranose was found and the intramolecular hydrogen bond was formed by hydroxide radical.

OBJECTIVE: To observe the effect of electro-acupuncture of Neiguan (PC 6) on mean systemic arterial blood pressure and plasmic concentrations of NO and TNFalpha in endotoxin shock rats. METHODS: The model of endotoxin shock was induced by lipopolysaccharide (1.5 mg/kg i.v.) and D-galactosamine (100 mg/kg i.p.). Aminoguanidine (100 mg/kg i.p.) and electro-acupuncture of bilateral Neiguan (PC 6) were administered. A catheter was inserted into the right subclavian artery to record the change of blood pressure and the blood was abstracted out and centrifuged to determine the NO and TNFalpha concentrations. RESULTS: Electro-acupuncture of Neiguan (PC 6) retrieved the blood pressure and reduced the plasmic NO and TNFalpha concentrations. CONCLUSION: Electro-acupuncture of Neiguan (PC 6) expresses an anti-endotoxin shock effect by repressing the plasmic NO and TNFalpha concentrations smoothly and retrieving the blood pressure stably.

[Objective] To observe the effect of serum containing Carapax et Plastrum Testudinis (CPT) in inducing the differentiation of mesenchymal stem cells (MSC) of adult rats into neurons. [Methods] The fifth generation of MSC were induced by the serum containing CPT (sero-CPT) and the expressions of neurofilament (NF) protein and glial fibrillary acidic protein (GFAP), the marks of neuron-like cells were detected by immunohistochemical method. [Results] The expression of NF protein was positive after the stimulation of sero-CPT and the expression reached the peak 12 hours after induction. [Conclusion] Sero-CPT can induce the differentiation of MSC into neurons in vitro.

[ Objective ] To investigate the mechanism of Jianxin Pinglu Pill (JPP) in counteracting arrhythmia. [Methods] Ninety rats were randomized into 3 groups: model (normal saline), high-dose JPP (1.6 g/kg) and low-dose JPP (0.8g/kg) . The treatment lasted 7 days. Rat models with myocardial ischemia /repermsion were established by blocking left coronary artery. Ⅱ lead of electrocardiogram was recorded, connexin 43 (Cx 43) was detected by immunohistochemical assay. The effect of JPP on incidence of arrhythmia, death rate and the expression of connexin 43 in the myocardium was observed. [Results] JPP could decrease the incidence of arrhythmia and death rate obviously and enhance the expression of connexin 43 in myocardium. [ Conclusion ] JPP can counteract the damage of myocardial ischemia /reperfusion to enhance Cx 43 expression, and this is one of its possible mechanism.

Objective To observe the effect of alteplase intra-arterial thrombolysis combined with continuous monitoring of transcranial Doppler (TCD) in patients with acute cerebral infarction. Methods Fifty-four patients with acute cerebral infarction were divided into TCD group and control group by random digits table method. The patients of 2 groups were given alteplase intra-arterial thrombolysis, and the patients of TCD groups were combined with continuous monitoring of TCD. The degree of clinical neurologic impairment was evaluated by National Institutes of Health stroke scale (NIHSS) and Barthel index (BI). The condition of vascular recanalization was evaluated by the change of blood flow signal of TCD. CT was performed within 1 d after thrombolysis to detect intracranial hemorrhage and other adverse reaction. The degree of clinical neurologic impairment, blood vessel recanalization rate after thrombolysis and recanalization time and incidence of intracranial hemorrhage and other adverse reaction were compared between 2 groups. Results There were no statistical difference in NIHSS score before thrombolysis and 1 h after thrombolysis between 2 groups (P>0.05). The NIHSS scores 7 and 30 d after thrombolysis in TCD group were significantly lower than those in control group:(8.1 ± 4.9) scores vs. (12.1 ± 4.2) scores and (6.9 ± 3.1) scores vs. (10.9 ± 3.9) scores, there were statistical differences (P<0.05). The BI scores 7 and 30 d after thrombolysis in TCD group were significantly higher than those in control group: (78.5 ± 13.8) scores vs. (60.8 ± 12.4) scores and (82.6 ± 13.5) scores vs. (63.2 ± 12.9) scores, and there were statistical differences (P<0.05). The blood vessel recanalization rate 24 h after thrombolysis in TCD group was significantly higher than that in control group: 70.4% (19/27) vs. 44.4% (12/27), the recanalization time after thrombolysis was significantly shorter than that in control group: (36.4 ± 9.5) min vs. (58.5 ± 12.4) min, and there were statistical differences (P<0.05). No adverse reaction like intracranial hemorrhage and so on was found by CT detection within 1 d after thrombolysis in the 2 groups. Conclusions Alteplase intra-arterial thrombolysis combined with continuous monitoring of TCD in acute cerebral infarction has good curative effect, and no obvious adverse reaction.

Objective To investigate the effect of Plastrum Testudinis on neural stem cell after focal cerebral ischemia.Methods Rat models of focal cerebral ischemia were established by inserting nylon thread into the anterior cerebral artery.Immunohistochemical method was used to detect the expression of the neuroepithelial stem cell protein(Nestin),neurologic deficit and histological features were also observed.Results Plastrum Testudinis can decrease the neurologic dificit score and promote the expression of Nestin.Conclusion Plastrum Testudinis can alleviate the neurological symptoms and promote the proliferation of neural stem cell after focal cerebral ischemia.

Objective: To explore the effect of Plastrum Testudinis on the three subtypes of NOS (nNOS,iNOS,eNOS) expression in rats with focal cerebral ischemia. Methods:Rat model of focal cerebral ischemia was set up by inserting nylon thread into the internal carotid artery . The expressions of nNOS, iNOS and eNOS was detected by immunohistochemical method to observe the effects of Plastrum Testudinis. Results: Expressions of nNOS and iNOS in cerebral cortex and caudate-putamen of Plastrum Testudinis treatment group were significantly lower than those of model control group (P

Objective To observe the effects of Niupo Zhibao pellet, a compound traditional Chinese herbal medicine,on helper T cell(Th cell)expression in rats with endotoxin-induced rapid pulmonary fibrosis (RPF). Methods Two hundred and twenty-four Wistar rats were randomly divided into four groups:normal control,model,dexamethasone(DXM)and Niupo Zhibao pellet(NW)groups(each n=56). By using endotoxin three-hit regimen,the RPF model was established. Three days before and 7 days after the establishment of models in NW group,they were administered with the pellet by intragastric feeding,50 mg/100 g of distilled water twice a day,a total of 10 days medication. Rats in DXM group received DXM intraperitoneal injection,3.0 mg/kg once a day for consecutive 7 days. Rats in normal control group were administered with the same volume of distilled water by intragastric administration. On 1,3,7,9,14,21,28 days after the administration,blood and bronchoalveolar lavage fluid(BALF)specimens were collected. The contents of γ-interferon(IFN-γ)and interleukin-4(IL-4) in the serum and BALF were investigated by enzyme-linked immunosorbent assays(ELISA). The lung tissue was stained by hematoxylin-eosin(HE)and Van Gieson respectively. Morphological changes in lung tissue were observed under light microscope. Results HE staining showed that the pulmonary interstitial tissues of rats in model group were thickened,there was a large amount of inflammatory exudates,and micro thrombi were seem in alveolar space. The pulmonary interstitial tissues in rats of DXM group were thickened too,but the inflammatory exudate in alveolar space was much less. Compared to rats in DXM group,the rats in NW group had slightly more inflammatory exudate and their pulmonary interstitial tissues were basically normal. Van Gieson staining showed that the expression of collagen fiber in model group was obvious,that of DXM group less than the former one but not significant,while that in the NW group was reduced markedly. ELISA assays demonstrated that the levels of IFN-γin BALF and serum in model group were increased significantly,those in DXM group were lowered to the minimum on the 7th day,raised to the levels before treatment on the 14th day,and gradually declined on the 21st day. In the NW group,the IFN-γwas consistently at a high level,and then gradually declined at a slow rate. After 7 days of drug administration,the IFN-γ levels in BALF and serum at various time points in NW group were obviously higher than those in model and DXM groups〔BALF(ng/L):140.47±4.22 vs. 149.23±8.35,90.67±6.65;serum(ng/L):140.47±4.15 vs. 100.43±11.05,99.35±7.85,P<0.05 or P<0.01〕. The levels of IL-4 in BALF and serum in each group increased significantly,reached to their maximum on the 7th day,and then gradually decreased. After the drug administration for 28 days,those levels in NW group were obviously lower than those in model and DXM group〔BALF(ng/L):6.60±1.05 vs. 7.20±1.25,8.55±1.05,serum(ng/L):6.75±1.05 vs. 7.21±1.25,8.25±1.15,P<0.05 or P<0.01〕. Conclusion Niupo Zhibao pellet can suppress inflammation,ameliorate injury of lungs and inhibit lung fibrosis by promoting IFN-γsecretion,restraining IL-4 secretion and adjusting the imbalance of Th cells.

BACKGROUND: Bone marrow-derived mononuclear cells (BM-MNCs) hold the potential of differentiating into osteoclasts. Polygonatum sibiricum polysaccharide (PSP) may inhibit the differentiation of BM-MNCs into osteoclasts and it is expected to become a new drug for the treatment of osteoporosis. OBJECTIVE: To investigate the effect of PSP on the differentiation of mouse BM-MNCs into osteoclasts induced by receptor activator of nuclear factor kappa-B ligand (RANKL) and bone resorption in vivo. METHODS: Mouse bone marrow-derived macrophages cultured in vitro, the effect of macrophage colony stimulating factor and PSP (5, 10, 20, 40, 80,160, 320, 640, 1280, 2560 mg/L) on the proliferation of mouse BM-MNCs was detected by cell counting kit-8 assay to determine the PSP concentration range; the mouse BMMs were cultured and induced in DMEM medium containing macrophage colony stimulating factor, RANKL and 5, 10, 20, 40, 80,160, 320, 640 mg/L PSP, respectively; those cultured without PSP served as control group. The morphological changes of cells were observed under an inverted microscope.; the number of osteoclasts was detected by tartrate-resistant acid phosphatase staining; the mRNA expression levels of osteoclast-related genes including tartrate-resistant acid phosphatase, matrix metalloproteinase-9, cathepsin K, and nuclear factor of activated T cells c1 were evaluated by quantitative real-time PCR. A mouse model of calvarial osteolysis induced by lipopolysaccharide was established to receive PSP intervention, and then micro CT scanning, three-dimensional reconstruction and relevants software were used for quantitative analysis of bone volume/volume percentage, trabecular number, trabecular bone spacing and thickness. The number of osteoclasts was identified by tartrate-resistant acid phosphatase staining and quantitative analysis of bone resorption area was conducted. RESULTS AND CONCLUSION: Compared with the control group, the concentration of PSP below 640 mg/L showed no significant effect on the proliferation of BMMs (P > 0.05). Different concentrations of PSP (40-640 mg/L) significantly reduced the number of osteoclasts, osteoclast differentiation and maturation, and the mRNA expression levels of tartrate-resistant acid phosphatase, matrix metalloproteinase-9, cathepsin K, and nuclear factor of activated T cells c1 TRAP, MMP-9, CtsK and NFATc1 (P < 0.05). Compared with lipopolysaccharide, PSP could effectively alleviate the lipopolysaccharide-induced calvarial osteolysis, and the bone volume/volume percentage, trabecular number, and trabecular bone spacing were significantly decreased (P < 0.05); additionally, the number of osteoclasts and the area of bone resorption were decreased significantly (P < 0.01). To conclude, PSP can inhibit the differentiation and maturation of mouse BMMs to osteoclasts and alleviate lipopolysaccharide-induced calvarial osteolysis.

To observe the effects of Niupo Zhibao Pellet, a compound traditional Chinese herbal medicine, on high-mobility group box-1 protein (HMGB1) expression in lung tissues of rats with endotoxin shock.