Objective: To observe the main pharmacodynamic effects of Yinaoning (YNN) on acute blood stasis rats and cerebral ischemia rats. Methods: Rats were prevented by the oral administration of YNN (0.4375～ 1.75g/kg once daily for 7 days. The acute blood stasis model and cerebral ischemaed model were used. Results: YNN could decrease the blood viscosity and plasma viscosity, shorten the length of in vitro thrombus, abate the wet and dry weights of thrombus in the acute blood stasis model group. It decreased the brain index significantly in the cerebral ischemia model group. The high dose of YNN could decrease the level of Evans blue obviously and reduce the degeneration of cranial nerve cells. These effects were similar to those of YNN Tablets and were dose dependent. Conclusion: YNN is effective for acute blood stasis rats and cerebral ischemaed rats.

Background Animal model of fungal keratitis is an available tool to the experimental study of the pathogenesis mechanism of fungal keratitis. Current modeling methods of fungal keratitis include corneal scratching, corneal stroma injection and corneal surface lens methods. But these methods still have their own shortages. Objective This experiment was to create a fungal keratitis animal model by modifying corneal surface lens method. Methods Modified animal models of fungal keratitis were created by modified corneal surface lens method in 12 general adult New Zealand white rabbits. The filter papers soaked 108 spores / ml or A106spores / ml of spergillus fumigatus suspension were attached on the de-epithelial cornea surface and fixed with contact lens and tarsorrhaphy for 2 days, and the filter paper with physiological saline was used as control group. The symptoms of anterior segment were examined under the slit lamp in 3 ,7 and 14 days after surgery and scored based on the criteria of Dong. Corneal scraping was stained with 10% potassium hydroxide and calcofluor white stain to observed mycelium under the fluorescence microscope. Corneal tissue sections were examined by hematoxylin-eosin staining and periodic acid Schiff staining under the light microscope. The use of animal followed the Standard of Association for Research in Vision and Ophthalmology. Results Fungal keratitis models were successfully established in 6 eyes and 4 eyes in 108 spores/ml group (6/6) and 106 spores/ml group respectively. The symptom was more severer and score was higher in the eyes of 108 spores/ml group than that in 106 spores/ml group. At 3 and 7 days after surgery,the symptom scores of fungal keratitis models were higher than those of control group from 3 through 7 days with the statistically significant difference (P<0. 01) and the symptom scores of 108 spores/ml group were significantly higher than those of 106 spores/ml group (P<0. 01). At 14 days after surgery, the symptom scores of 108 spores/ml group were still higher than those of control group (P<0. 05). Fungal hyphae was seen in the corneal scrapes in 108 spores/ml group and 106 spores/ml group respectively from 3 through 7 days after surgery. Inflammatory cell infiltration, stroma cells necrosis and fungal hyphae were presented in 108 spores/ml group, and the corneal neovascularization could be observed in 108 spores / ml group 14 days later. Fungal culture revealed the positive outcome in both 3 and 7 days after surgery in 108 spores/ml group,but in 106 spores/ml group,the positive result was only in the 3rd day. Conclusion Modified corneal surface lens method is more feasible and sample in the model of Aspergillus keratitis. This animal model of Aspergillus keratitis is practical for the further study of fungal keratitis.

ObjectiveTo evaluate central nervous system function of patients with coronary cardiac diease by short latency somatosensory evoked potentials (SSEP).MethodsThe cerebral and spinal somatosensory evoked potentials were recorded by stimulating median nerve in 43 patients with coronary cardiac disease but without apparent nervous symptoms and 14 healthy control subjects.ResultsThe lactency periods and central conductive time of N13, N20 and P25 wave were significantly prolonged in patients with myocardial infarction (MI) or angina pectoris (AP) when compared with normal controls (P<0.05～0.001). The lactency periods and central conductive time of N20 and P25 wave recorded in MI patients were longer than those recorded in AP patients (P<0.01～0.001).ConclusionThe subclinical nervous damages in the central somatosensory pathway from spinal cord to cerebral cortex is present in patients with coronary cardiac disease especially myocardial infarction.

Objective To observe effects of ginsenoside Rb1 on phosphorylation of microtubule-associated protein 2 (pMAP-2) in hippocampus and amygdala of depressive model rats. Methods Thirty male Wistar rats were randomly divided into control group, model group and treatment group. The depression rat model was produced by giving chronic unpredicted mild stress (CUMS). Treatment group was given daily intragastric administration of ginsenoside RB 1 (1 g/mL crude drug, 1 mL/100 g body weight) for 22 days during modeling. Western blot assay was used to detect expressions of MAP-2 and pMAP-2 protein, and real-time PCR was used to detect expressions of pMAP-2 mRNA respectively. Results The expressions of pMAP-2 protein and mRNA in hippocampus and amygdala were significantly lower in model group than those of control group (P < 0.05). The expressions of pMAP-2 protein and mRNA were significantly higher in treatment group than those of model group (P<0.05). Conclusion Ginsenoside Rb1 can play anti-depression role by inhibiting the phosphorylation of MAP-2 in rats.

OBJECTIVETo observe the clinical effect of Chinese medical (CM) syndrome differentiation based Chinese herbs and recombinant human tumor necrosis factor receptor II-antibody fusion protein (etanercept) for treating ankylosing spondylitis (AS) patients.

METHODSTotally 35 AS patients were treated with syndrome differentiation based Chinese herbs and etanercept. Reinforcing Shen and strengthening Du channel, activating meridians to stop pain was principle used in syndrome differentiation based treatment. Etanercept was subcutaneously injected, 25 mg each time; twice per week for the first three months and once a week for the latter three months. The clinical efficacy was evaluated after 3 and 6 months of treatment. Meanwhile, ASAS20 and ASAS50 standards arriving rates were also observed. Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), Bath Ankylosing Spondylitis Functional Index (BASFI), visual analog score (VAS) for spine pain, VAS for night pain, patient global assessment (PGA), VAS for physician global assessment, CM syndrome score, finger-ground distance, thoracic activity, tragus-wall distance, lumbar scoliosis, cervical rotation, Schober improved test, erythrocyte sedimentation rate (ESR), and C-reactive protein (CRP) were observed before treatment, 3 and 6 months after treatment.

RESULTSCompared with before treatment, BASDAI, BASFI, VAS for spine pain, night pain, physician global assessment, PGA, CM syndrome score, finger-ground distance, thoracic activity, tragus-wall distance, lumbar scoliosis, Schober improved test, ESR, and CRP all decreased after 3 and 6 months of treatment, with statistical difference (P < 0.05). Cervical rotation also decreased after 6 months of treatment, with statistical difference (P < 0.05). Compared with 3 months of treatment, total effective rate of CM syndrome, ASAS20 and ASAS50 standards arriving rates increased after 6 months of treatment, with statistical difference (P < 0.05). There were statistical differences in all indices mentioned above between after 3 months of treatment and after 6 months of treatment (P < 0.05).

CONCLUSIONSyndrome differentiation based Chinese herbs combined etanercept could alleviate inflammatory reaction favorably, control the progression of active AS, and improve joint functions.

Disease Progression ; Drugs, Chinese Herbal ; therapeutic use ; Etanercept ; therapeutic use ; Humans ; Pain ; prevention & control ; Pain Management ; Spondylitis, Ankylosing ; drug therapy ; Treatment Outcome

With the in-depth research of blood circulation and blood stasis, many scholars gradually carried out research and exploration the pathophysiological basis of blood stasis of the ankylosing spondylitis (AS), from the blood rheology, microcirculation, platelet activation, vascular endothelial function and so on. Some progress that been made were summarized below.

The paper is to establish a method for simultaneous determination of 5 kinds of alkaloids in ephedra and poppy which are in Kechuanning tablets. Solid-phase extraction (SPE) was adopted in pretreatment, and a UPLC method with 2 different wavelengths had been developed: 210 nm for the detection of morphine, codeine phosphate, ephedrine hydrochloride and pseudoephedrine hydrochloride, and 251 nm for papaverine hydrochloride. The column used was Acquity UPLC BEH C18 (100 mm x 2.1 mm ID, 1.7 microm) with linear gradient elution using acetonitrile and 0.1% phosphoric acid. The flow rate was 0.4 mL.min-1, and the column temperature was 30 degrees C. The linear response range was 0.375 0 - 12.50 microg.mL-1 for morphine, 0.064 32 - 2.144 microg.mL-1 for codeine phosphate, 0.030 06 - 1.002 microg.mL-1 for papaverine hydrochloride, 1.126 - 37.52 microg.mL-1 for ephedrine hydrochloride, 0.287 8 - 9.592 microg.mL-1 for pseudoephedrine hydrochloride (r = 0.999 7). The average recoveries of these compounds were 99.26%, 100.6%, 95.29%, 100.1% and 97.48%, respectively. This is a more reasonable and credible method of quality control for Kechuanning tablets.

Contents in orthopaedic are independent relatively and finding out the internal relations during them is helpful to improving the effect of orthopaedic teaching.Transfer theory is important tools for clinical teaching practice,and finding out the common characteristics between the orthopaedic chapters is primary for the theory.This research focuses on the following fields:fractures,nerve injury,infection,tumor and deformity.After the common characteristics between these chapters were analyzed and discussed,we concluded that the transfer theory is helpful in orthopaedic teaching practice,especially for students' comprehension and memory,but still we should avoid some negative effects in teaching process.

Objective: To explore the role of anti-?? T cell receptor(TCR) and anti-CD80 monoclonal antibodies(mAbs) combined with donor bone marrow cells(BMCs) infusion in the induction of murine skin allografts tolerance.Methods: On day 0,2?10~8 BMCs of BALB/c mice were injected into recipient C57BL/6 mice via the tail vein,meanwhile,an intraperitoneal injection of TCR?? mAb(500 ?g) was given.On day 2,CD80 mAb was administered intraperitoneally.Skin grafting was performed on day 6.Delayed type hypersensitivity(DTH),mixed lymphocyte reaction(MLR),IL-2 reverse assay of MLR,adoptive transfer assay and chimerism detection were performed at different time points and tolerance mechanisms were investigated.Results: The mean survival time(MST) of BALB/c skin allografts in C57BL/6 recipients that were treated by anti-TCR?? and anti-CD80 mAbs combined with donor BMCs infusion was 70 days.DTH and MLR assay indicated that the tolerant mice displayed significant hyporesponsiveness.The result of IL-2 reverse test showed that clone anergy was probably involved in the formation of tolerance in the tolerant C57BL/6 mice.In vivo and in vitro adoptive transfer assay,suppressive activity in the spleens of tolerant C57BL/6 mice was observed.Chimerism existed in both the thymus and spleen of the tolerant C57BL/6 mice.The chimerism level gradually declined with time.Conclusion: Treatment of anti-TCR?? and anti-CD80 mAbs combined with donor BMCs infusion can successfully induce a long-term tolerance in BALB/c mice to C57BL/6 skin graft.Multiple mechanisms,including clone anergy,suppressor cells and chimerism are involved in the tolerance.

The influence of short hairpin RNA (shRNA)-mediated osteopontin (OPN) gene silencing on the proliferation and invasion of human renal cancer ACHN cells was investigated. Four types of OPN shRNA recombinant plasmids were constructed and RT-PCR assays were used to screen the most highly functional shRNA recombinant plasmids, which were transferred into the cultured ACHN cells by Lipofectamine 2000. The cells transfected by shRNA expression vectors (ACHN/OPN) were visualized under an inverted microscope and screened by G418. Untreated cells (ACHN) and cells transfected by mock vectors (ACHN/Vect) were used as control groups. The expression levels of OPN mRNA and protein were detected by real-time PCR and Western blot respectively. The cell cycle and ratios of apoptotic cells were assessed by flow cytometry. MTT method was used for drawing the growth curve and observing cell proliferation in vitro. The abilities of migration and invasion in three groups were measured by Transwell chamber test. The expression levels of matrix metalloproteinase (MMP)-2 and MMP-9 in three groups were examined by Western blot. Our results showed that the recombinant plasmid could be successfully transferred into ACHN cells by LipofectamineTM 2000. Compared with untreated cells, the expression levels of OPN mRNA and protein in ACHN/OPN cells were decreased by 59.68% and 76.42%, respectively (P<0.05), ACHN/OPN cells were blocked in S phase and apoptotic ratio increased significantly (P<0.05), however, no significant differences were found between ACHN/Vect and ACHN. Recombinant plasmid significantly attenuated expression levels of MMP-2 and MMP-9 proteins and suppressed the proliferation, migration, and invasion of ACHN cells. This study suggested that OPN may play an important role in the growth and invasion of human renal cancer ACHN cells, and these processes are correlated with the activations of MMP-2 and MMP-9. Our data provided preliminary experimental evidence for the feasibility of RNA interference technology in gene therapy of human renal cancer.