Objective To investigate the best conditions of extraction techniques for compound Tianqi Yizhi granule and to establish its quality standards. Methods The best conditions of extraction techniques for compound Tianqi Yizhi granule were optimized by orthogonal experiments, with the yield of total saponins as the index. The qualitative analysis of Panax Notoginseng and Ginseng with Thin-Layer Chromatography and determination of total saponins with UV-spectrophotometry was made. Results The optimal condition of total saponins was that the microwave extraction for 3 times and extracting for 30 minutes, the material-water rate of 1∶10. In TLC procedure, the control material and the test sample showed the same colored spots, while the negative control sample showed no interference. Ginsenoside Re showed a good linear relationship at the range of 40～200 ?g, Y =250.83X -0.017, r =0.997 2. The accuracy and precision were accord to the standards. Conclusion The extraction techniques and the quality standards are feasible.

Objective:To investigate the effect of GRP94 gene expression down-regulation on the proliferation in human lung carcinoma A549 cells and possible mechanism.Methods:Expression of GRP94 was examined in 4 lung carcinoma cells and two lung bronchial epithelial cells by qRT-PCR and Western blot.Chemically synthesized siRNA targeting GRP94 gene and transfected into A549 cells used by Lipofectamine2000.The mRNA and protein expression levels of GRP94,c-myc and cyclinD1 were detected by qRT-PCR and Western blot.CCK8 assay and Colony formation assay were used to detect the effect of specific GRP94 siRNA on cell proliferation.Results:The levels of GRP94 expression in the four lung carcinoma cells were significantly higher than that in two lung bronchial epithelial cells (P<0.05).GRP94 siRNA significantly downregulated the levels of GRP94 mRNA and protein in A549 cells, and the downregulation evidently suppressed cell proliferation.However, The c-myc and cyclinD1 expression levels in A549 cells in siRNA-GRP94 group was significantly lower than that in siRNA-NC group (P<0.05).Conclusion:The levels of GRP94 expression in lung carcinoma cells were significantly higher than that in normal lung epithelial cells.Knockdown GRP94 expression significantly inhibit the level of lung carcinoma A549 cells proliferation,which may act via influencing the expression of c-myc,cyclinD1.

Objective:To research the anti-inflammatory effect of LL-37 in mycobacterium tuberculosis ( Mtb ) infected-macrophages,as well as its influence on the secretion of inflammatory cytokines. Methods:( 1 ) THP-1 cells were cultured and incubated with phorbol 12-myristate 13-acetate (PMA) to transform into an adherent macrophage-like state (macrophage,Mφ). Then the THP-1 cell derived macrophages were infected with mycobacterium tuberculosis,and then stimulated with different concentrations of LL-37. (2)The experiment was divided into following groups: ① Control group:THP-1+normal saline (NS);② Mtb group:THP-1+Mtb;③Mtb+LL-37 5 μg/ml group:THP-1+Mtb+5 μg/ml LL-37;④Mtb+LL-37 10 μg/ml group:THP-1+Mtb+10 μg/ml LL-37;⑤Mtb+LL-37 20 μg/ml group:THP-1+Mtb+20 μg/ml LL-37. (3) The mRNA expression levels of IL-12p40,TNF-α,IL-4,and IL-10 will be determined by Real-time PCR respectively at 6,12,24 and 48 hours. The secreted levels of IL-12p40,TNF-α,IL-4,and IL-10 will be determined by ELISA analysis respectively at 6,12,24 and 48 hours. Results:The mRNA expression levels and secreted levels of IL-12p40,TNF-α,IL-4 and IL-10 were increased in Mtb group than those in control group. The mRNA expression levels and secreted levels of pro-inflammatory cytokines IL-12p40 and TNF-α were decreased in the LL-37 groups than those in Mtb group. However,anti-inflammatory cytokines IL-4 and IL-10 mRNA expression levels and secreted levels were increased in the LL-37 groups than those in Mtb group. Conclusion:Exogenous LL-37 inhibited the secretion of inflammatory cytokines in macrophages during mycobacterium tu-berculosis infection. The effect is related to the concentrations of LL-37 and the stimulated time of macrophages which were infected with mycobacterium tuberculosis. The results will provide new insights into the treatment of Mtb infection.

Objective To investigate the role of ephrin A genes in the development of oxygen-induced retinal neovascularization (OIR) in mice. Methods The OIR model was established by oxygen induction in new born C57BL/6J mice. Reversed transcript - polymerase chain reaction (RT-PCR) was used to measure the expression levels of ephrin A1-A5 in retinas of mice in experimental and normal control group. Results All of the ephrin A family genes expressed in normal retinas. Ephrin A1 mRNA was significantly higher in OIR group (t= 3.19, P = 0. 019); ephrin A2 mRN A was higher in the 15-day-old OIR retinas (t= 3. 71,P=0. 033) ; ephrin A3-A5 mRNA decreased or disappeared in 12- and 13-day-old RNV mice, and increased in 15-day-old OIR mice. Conclusion Ephrin A genes are involved in the development of retina and OIR.

Objective To prepare a harmine hydrochloride liposome formulation with high encapsulation efficiency. Methods Preformulation investigation was carried out to obtain the drug physicochemical properties such as solubility and lgD in buffers of different pH value. Harmine hydrochloride was encapsulated into liposomes by active loading method. Encapsulation efficiency of liposomes was determined after the free drug was separated from liposome by ultrafiltration. The influence factors on the encapsulation efficiency including drug-lipid weight ratio, incubation temperature, pH value of external water phase were investigated. Results As the pH value increasing, the solubility of harmine hydrochloride was decreased, while the apparent oil-water distribution coefficient was increased. By active loading method, the encapsulation efficiency could be over 80% when the drug to lipid weight ratio was under 1∶5. The pH gradient between intervesicle and intravesicle obviously influenced the encapsulation efficiency, while incubation temperature had little effect on encapsulation efficiency. Conclusion Active loading is suitable for preparing harmine hydrochloride liposome with high encapsulation efficiency.

Objective To investigate the clinical value of olfactory functional magnetic resonance imaging in early diagnosis of Alzheimer's disease (AD).Methods 43 patients with mild cognitive impairment were selected,in the same period,49 cases of patients with AD were selected as the AD patients group,and 53 normal populations were selected as the control group.The neuropsychological of all cases were assessed by using MMSE scale,MoCA scale and CDR Scale.Siemens 3.0T MRI machines were used according to event -related design approach for olfactory cor-tex conducted fMRI scans.The matlab7.0 and SPM8 data preprocessing tools were used to image analysis.The prima-ry olfactory cortex volumes and activation voxels numbers of the three groups were compared.The correlations of acti-vation voxels numbers and primary olfactory cortex volumes and neuropsychological scores were analyzed.Results The primary olfactory cortex volumes of the control group,patients with mild cognitive impairment and AD patients were (3 846 ±517)mm3 ,(2 863 ±367)mm3 and (2 214 ±283)mm3 ,respectively,pairwise comparison,the control group >mild cognitive impairment >AD patients,the differences were statistically significant (compared with the con-trol group,t =16.835 and 34.716,compared with mild cognitive impairment group,t =19.753,P <0.05).The acti-vation voxels numbers of the control group,patients with mild cognitive impairment and AD patients were (193 ±84), (104 ±52)and (51 ±27),pairwise comparison,the control group >mild cognitive impairment >AD patients,the differences were statistically significant (compared with the control group,t =47.916 and 72.954,compared with mild cognitive impairment group,t =37.382,P <0.05).Partial correlation analysis showed that the activation voxels num-bers of the three groups were positively correlated with primary olfactory cortex volume,MMSE score and MOCA score (r =0.397,0.462 and 0.494,all P <0.05).Conclusion Olfactory fMRI in patients with AD might reflect the changes in the entorhinal cortex caused by pathological changes.It could provide clues and information for the early diagnosis of AD.

OBJECTIVE:To provide reference for improving pharmaceutical care in the hospital. METHODS:A total of 712 clinical pharmacists were selected from 38 third grade class A hospitals in 17 provinces and cities by convenient sampling to investi-gate the ability of pharmacectical care and influantial factors. Single and multiple factor Logistic regression analysis were conducted for the results. RESULTS:The single factor analysis showed that there was significant difference in the communication between clinical pharmacists and patients or medical staffs,the listening attitude of the patients,attitude towards job prospects,understand-ing about the connotation of clinical pharmacy,the timeliness of drug supply,smoothness degree of channel,the timeliness of coordi-nation,smooth development of pharmaceutical care(P<0.05). The multiple factor analysis showed that due to good communication with the doctor [OR=1.819,95%CI(1.469,2.253)],pharmacists could understand definition and connotation of pharmaceutical care [OR=2.263,95%CI(1.722,2.973)],and the male pharmacists [OR=0.558,95%CI(0.382,0.813)] with positive attitude to the pharmacists [OR=1.506,95%CI(1.309,1.733)] believed to provide good pharmaceutical care. CONCLUSIONS:This investigation can reflect the present situation of hospital pharmaceutical service in China to a certain extent. It is suggested to improve pharmaceuti-cal care ability of pharmacists through strengthening resource disposition and the cooperation between physicians and pharmacist.

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Migraine is one of the common and frequently encountered diseases. The study proves that 5-hydroxytryptamine (5-HT) receptor, plays an important role in the occurrence of migraine. Rat striatum was used for preparation of the cell membrane stationary phase (CMSP) in our experiments. The cell membrane chromatography (CMC)-offline-HPLC system was applied to specifically recognize the components from the drug pair of Chuanxiong Rhizoma and Angelicae Dahuricae Radix, which interact with the receptors on CMSP. The dissociation equilibrium constant (KD) was measured in a rat striatum/CMC system, performed by continuously pumping sumatriptan, a 5-HT1D agonist, ranging from 2.42 x 10(-8) to 4.84 x 10(-7) mol · L(-1) through a CMC column, and the capacity factors (k') were recorded. The KD value obtained from the model was (4.59 ± 0.33) x 10(-6) mol · L(-1) for imperatorin, and the rat model of migraine induced by nitroglycerin was applied to validate the pharmacological effects of the drug pair. The results indicated that the CMC method could be a quick and efficient way for characterizing the drug-receptor interactions in vitro.
Angelica ; chemistry ; Animals ; Cell Membrane ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drug Evaluation, Preclinical ; methods ; Drugs, Chinese Herbal ; chemistry ; Male ; Migraine Disorders ; drug therapy ; Rats ; Rats, Sprague-Dawley ; Receptor, Serotonin, 5-HT1D ; chemistry ; Serotonin Receptor Agonists ; analysis

Forty-two patients with type 2 diabetes (DM group),23 type 2 diabetic patients with unstable angina pectoris (DM + UAP group),and 36 healthy subjects (control group) were enrolled in this study.Serum samples were collected for determining serum concentrations of vascular endothelial growth factor (VEGF),Angiopoietin (Ang)-1,Ang-2,angiogenin,angiostatin,basic fibroblast growth factor,and platelet-derived growth factor-BB using protein array technology.The results showed that there were no significant differences in serum concentrations of all 7 angiogenic factors between control group and DM group.Whereas,serum concentrations of V EGF and Ang-2 were significantly increased in DM + UAP group compared with control group [(3 532.10 ± 1 813.72vs 2 444.50 ± 1 152.21) pg/ml,(286.90-± 217.01 vs 171.92 ± 106.63) pg/ml,both P＜0.01].Pearson's correlation analysis revealed that serum concentrations of all 7 angiogenic factors were not related to HbA1C,fasting and 2 h postprandial plasma glucose,triglycerides,high density lipoprotein cholesterol,as well as low density lipoprotein cholesterol.