OBJECTIVE:To optimize the extraction technology of nucleoside from Codonopsis pilosula. METHODS:The ultra-sonic extraction was used to extract nucleoside from C. pilosula. The extraction technology was screened and optimized by single factor and orthogonal test with ethanol concentration (including water),extraction time,solid-liquid ratio and extraction times as factors using the total contents of cytidine,uridine,guanosine and adenosine as index. Validation test was conducted. RESULTS:The optimal extraction technology was that pure water was extraction solvent,extracting for 45 min,with material-liquid ratio of 1∶20 (g/ml),extracting for 2 times. Total amount of 4 kinds of nucleosides from in 100 g C. pilosula was (17.01 ± 0.005) mg (RSD=0.68%,n=3). CONCLUSIONS:Optimized extraction technology of nucleoside from C. pilosula is stable,feasible,rapid in operation and cost-saving.

Objective:To investigate the anti-proliferation and anti-metastasis effects and study the molecular mechanism of sinomenine in cell line(HepG2).Methods: HepG2 cells were cultured together with different treatment concentrations of sinomen-ine.The effect of sinomenine on inhibition of growth of HepG2 cells were determined by methyl thiazolyl tetrazolium(MTT) assay.The effect of sinomenine on inhibiting metastasis of HepG2 cells were determined by Transwell assay.The inhibitory effect of sinomenine on reverse transcriptase(RT) was studied using inhibitory kinetic method,on the basis,the reactive oxygen species(ROS) of HepG2 cells was monitored by indirect fluorescent labeling.The protein expressions of CASP3,CASP9,CAV1 and SOX2 were analyzed by Western blot experiment.Results: Sinomenine inhibited the proliferation and metastasis of HepG2 cells significantly.Sinomenine had a good inhibitory effect on the growth of HepG2 cells,half inhibitory concentration(IC50) was (15.35±2.43)μmol/L.Sinomenine was RT inhibitor,IC50 was (21.32±2.43)μmol/L.The Western blot showed that CASP3,CASP9 and CAV1 were up-regulated and SOX2 was down-regulated by the sinomenine treatment in HepG2 cells.Conclusion: The potential molecular mechanism of sinomenine suppresses proliferation and metastasis of HepG2 cells by up-regulation of CASP3,CASP9,CAV1 and down-regulation of SOX2.

Objective To observe the effect of acupuncture on cerebral palsy children. Methods90 children with cerebral palsy were divided into the observation group (acupuncture and comprehensive rehabilitation) and control group (comprehensive rehabilitation). They were assessed with Gross Motor Function Measure and the Gesell Development Schedules before and 3 months after treatment. ResultsThe total effective rate was 95% in observation group, and 80% in the control group (P<0.01). The score of gross motor function and Gesell adaptive development quotient improved more in the observation group than in the control group (P<0.01). ConclusionAcupuncture is efficacious on functional recovery in children with cerebral palsy

非经典信号通路IKKε和TBK1与恶性肿瘤密切相关，多种因素激活IKKε和TBK1通路，可引起NF-κB途径的激活， 导致肿瘤细胞的凋亡减少、细胞周期加快，促进肿瘤发生和发展。抑制IKKε和TBK1信号通路，可增加多种细胞凋亡因子的表 达，抑制肿瘤细胞增殖，促进肿瘤细胞凋亡，同时提高化疗和放疗的敏感性。因此，阻断IKKε和TBK1信号通路可有效治疗恶性 肿瘤，已有的实验证实有多种阻断IKKε和TBK1通路的药物均具有良好的抗肿瘤作用。

Objective To research a fast method on establishment of simplified model of the spine.Methods The 3D geometry of the spine is reconstructed using a multiview radiography reconstruction technique through the two plane X-rays by extracting the spinal anatomy information combined with the statistical geometrical information.Results The entire spine finite element modeling including the thoracic spine(T1-T12),lumbar spine(L1-L5) and the sacrum was reconstructed,and the analysis proved that lateral corrective force can correct scoliosis to a certain extent.Conclusion Because the method needs less time,and causes less radiation to patients,it can be brought much convenience and support to the clinical application.

Objective To assess the feasibility of K value of dynamic contrast enhanced MRI (DCE-MRI) in evaluation of anti-angiogenic effects on nude mice with orthotopic transplantation tumor model of gastric cancer.Methods Nude mice with orthotopic transplantation tumor model of gastric cancer were randomly assigned to two groups:Treatment group (n =15),mice were given apatinib intragastrically for 18 days (100 mg/kg),and control group (n=15),mice were given ddH2O2 in the same manner.After 18 days,DCE-MRI was performed and K value was measured.Then the tumors were dissected from the adjacent tissues in order to detect the microvessel density (MVD) and vascular endothelial growth factor (VEGF) expression levels.MVD and VEGF expression level were compared between treatment group and the control group.Results MVD (x2 =4.89,P＜0.05) and VEGF expression level (x2 =8.69,P＜0.01) of treatment group were much lower than those of control groups.The Kt value of treatment group was significantly lower than that of control groups ([0.63±0.05]/min vs [1.66±0.23]/min,t=17.05,P＜0.01).Conclusion The value of k in DCE-MRI can be utilized to assess the effects of apatinib on nude mice with orthotopic transplantation model of gastric cancer.

Objective To investigate the factors that influence the fermentation process of compound Chinese medicine and determining the optimum fermentation with single factor experiment and response surface methodology. Methods Through controlling the factors in the fermentation process of compound Chinese medicine (such as fermentation bacteria, fermentation time, fermentation temperature, inoculum amount, etc.), with its increase rate of total peak area as evaluation indicator, the alcohol extracts before and after fermentation were monitored and comparative evaluated by HPLC, and the optimum fermentation process was determined by response surface methodology. Results The fermentation process optimized by single factor experiment and response surface methodology was as follows:SZ-2 strain served as the fermentation bacteria, temperature was 33 ℃, inoculum amount was 4%, and time was 3.5 d, the average increase rate of total peak area was 31.24%. Conclusion HPLC can be used to identify and evaluate the fermentation of compound Chinese medicine under the different factors, and to clarify the optimal fermentation process by response surface methodology, which provide reference for the development of fermentation process.

Objective To explore the effect of erlotinib in patients with non-small-cell lung cancer (NSCLC)in stage ⅢA before operation and the relation with the rate of resection,good operability and postoperative complications.Methods 31 patients with NSCLC in stage ⅢA in group A were treated with erlotinib before operation;34 patients with NSCLC in stage ⅢA in group B only were treated with operation.Results The condition of 64.5%patients were improved.The operability in group A was more than that in group B(P=0.008),the good operability in group A was more than that in group B(P=0.011),the postoperative complications do not have statistical significance (P=0.07).Conclusion The erlotinib can increase the rate of resection in patients with NSCLC in stage ⅢA,and increase good operability in patients with NSCLC in stage ⅢA,but not increase the postoperative complications.

Objective:To investigate the effect of long-chain non-coding CDKN2B targeting miR-19 on the biological behavior of chronic myeloid leukemia cells and its mechanism.Methods: The expression of CDKN2B in different leukemia cells were detected by qPCR.Double luciferase reporter gene was used to detect the interaction between CDKN2B and miR-19.MTT proliferation assay and flow cytometry were used to detect the effect of CDKN2B on the proliferation and apoptosis of HL-60 cells.The changes of migration ability of leukemia HL-60 cells after overexpress of CDKN2B were detected by scratch test.The changes of invasion ability of leukemia HL-60 cells after silencing CDKN2B were detected by Transwell invasion assay.Scaling healing experiment and Transwell invasion assay were used to detect the effect of miR-19 on the migration and invasion of leukemia cells after silencing CDKN2B.The morphological changes of cytoskeleton microfilament microtubules after silencing CDKN2B were detected by phalloidin staining.Western blot was used to detect the expression of PI3K/AKT signaling pathway after silencing CDKN2B.Results: The expression level of CDKN2B was the lowest in leukemia cell HL-60.CDKN2B binds specifically to the 3′UTR of miR-19;overexpression of CDKN2B could inhibit the proliferation and enhance the apoptosis of leukemia HL-60 cells.Overexpression of CDKN2B can inhibit the invasion and migration of leukemia HL-60 cells.After overexpressed of CDKN2B,the cytoskeleton showed decreased pseudopodia and decreased exercise capacity.The expression of actin was down-regulated.The expression of PI3K/AKT pathway protein was down-regulated after overexpressed of CDKN2B.Conclusion: CDKN2B can target the regulation of miR-19 to regulate the biological behavior of leukemia cells.

AIM: To observe the protein expression of vascular endothelial growth factor 1 (VEGF-1) in pulmonary arterial endothelial cells and VEGF-1 gene expression in lung tissue in rats with hypoxia-induced pulmonary hypertension and treated with heparin. METHODS: Twenty four male adult SD rats were randomly divided into three groups (8 rats each): a control group (group A), a group with hypoxia for 4 weeks (group B) and a group with hypoxia for 4 weeks and injected with heparin to abdominal cavity simultaneously (group C). Mean pulmonary arterial pressure (mPAP), right ventricle hypertrophy index (RVHI) and vessel morphometry were measured. The morphology of pulmonary artery was observed by HE staining. The expression of VEGF-1 protein in pulmonary arterial endothelial cells was determined by immunohistochemistry. The level of VEGF-1 mRNA in lung tissue was measured by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: mPAP, RVHI, pulmonary artery remodeling parameters, VEGF-1 protein expression in pulmonary arterial endothelial cells and VEGF-1 gene expression in lung tissue of the three groups from high to low were group B, group C and group A. It was statistically significant when compared between either two groups of the three (P<0.01). Linear correlation analysis showed that VEGF-1 protein was positively correlated with pulmonary artery remodeling parameters (r=0.974, P<0.01), and VEGF-1 mRNA was positively correlated with VEGF-1 protein (VEGF 120 mRNA, r=0.919, P<0.01; VEGF164 mRNA, r=0.896, P<0.01). CONCLUSION: Heparin may down-regulate the expression of VEGF-1 at the levels of transcription and translation, resulting in the inhibitory effect on rats with hypoxia-induced pulmonary hypertension.