Objective To optimize the purification technology of total flavonoids from the Tripterospermi Chinensis Herba by macroporous resin. Methods The purification abilities of six kinds of macroporous adsorption resins for total flavonoids from Tripterospermi Chinensis Herba were studied with the adsorption and desorption rates as the index by static adsorption and desorption experiments. The column liquid concentration, adsorption rate, and the loaded amount were studied by using dynamic adsorption experiments. The optimal desorption technology was examined by orthogonal experiment. Results AB-8 macroporous resin was found to have good adsorption and desorption effects. The optimal purification conditions were as follows:the column liquid concentration was 5.285 mg/mL, adsorption rate was 2 BV/h;the loaded amount was 17.62 mg/mL. The sample was eluted by 10%ethanol with 4 BV and 50%ethanol of 5 BV at a flow rate of 4 BV/h. The purity of total flavonoids increased to 61.95% after the purification, and the yield was 87.28%. Conclusion This optimized process was stable, feasible and suitable for separation and purification of total flavonoids from Tripterospermi Chinensis Herba.

Objective To study the adsorption technology of piceid on macroporous resin. Methods The extraction condition of piceid, the types of macroporous resin, the concentration of the sample of Rhizoma Polygoni cuspidati extract, the flow velocity of samples during adsorption were investigated and the leak curve was drawed to ascertain the optimal sample quantity. Results Resin D101 gave a better adsorption performance in the following technological conditions:the concentration of the sample of Rhizoma Polygoni cuspidati extract was 0.7774 mg/mL, the flow velocity of sample during adsorption 3BV/h, the optimal adsorption capacity of the resin was 10.88 mg/mL. Conclusion The procedure was simple, feasible and can be applied to industrial production.

Objective To investigate the antitumor mechanism of astragaloside combine with cetuximab on the colon cancer cell line RKO of EGFR over expression through cell proliferation and autophagy. Methods The cell proliferation of colon cancer cell line RKO intervened by astragaloside with or without cetuximab was detected by 4- methyl- teerazolium (MTT). The expressive changes for protein of EGFR 、P62 and LC3 in RKO cells was detected by western bolt. Results MTT showed that 100 ug/mL astragaloside combined with 120 ug/mL cetuximab had significantly inhibiting effect on RKO cells of EGFR expression. Western blot showed that astragaloside can affect the expression of P62 and LC3 to suppress the occurrence ofautophagy. Conclusion In the vitro studies showed that astragalosidecan enhance the antitumor cell proliferation of cetuximab through inhibit autophagy in the treatment of colon cancer.

Objective To investigate the role of phosphatidyl-inositol 3-kinase-Akt (PI3k-Akt) signal pathway in the attenuation of ischemia-reperfusion (I/R) injury by sevoflurane preconditioning in isolated rat hearts. Methods Ninety-six adult male SD rats weighing 220-280 g were randomly divided into 6 groups ( n = 16 each): sham operation group (group S); I/R group; sevoflurane preconditioning group (group SP); wortmannin group (group W); dimethyl sulfoxide (DMSO) group (group D) and sevoflurane preconditioning + wortmannin group (group SW) . Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95%O2-5%C02 at 37 ℃ . The hearts were continuously perfused for 180 min in group S. After 15 min of equilibration, the isolated hearts were subjected to 30 min of ischemia followed by 120 min of reperfusion in SP, W, D and SW groups. Croups SP, W, D and SW received 10 min of perfusion with K-H solution containing 2. 4% sevoflurane, 100 nmol/L wortmannin, 20 μmol/L DMSO, and 2.4% sevoflurane + 100 nmol/L wortmannin, respectively, followed by 5 min washout before I/R. Eight hearts in each group were selected and HR, left ventricular end-diabetic pressure (LVEDP), left ventricular developed pressure (LVDP), and ± dp/dtmax were recorded at the end of equilibration and at 15 min of reperfusion, Myocardial tissues were obtained at 15 min of reperfusion for determination of apoptosis (by TUNEL) and phosphorylated Akt (p-Akt) expression (by Western blot) . Another 8 hearts were selected at 120 min of reperfusion for determination of myocardial infarct size by TTC staining. Result Compared with group S, LVDP and ± dp/dt,^ were significantly decreased and LVEDP was significantly increased in groups I/R, SP, W, D and SW, and myocardial p-Akt expression was up-regulated in groups I/R, SP and D ( P ＜ 0.05). Compared with group I/R, LVDP and ± dp/dtmax were significantly increased, LVEDP and apoptosis index were significantly decreased, myocardial p-Akt expression was up-regulated, and myocardial infarct size was significantly reduced in group SP (P ＜0.05) . Conclusion Activation of PI3K-Akt signal pathway is involved in the attenuation of I/R injury by sevoflurane reconditioning in isolated rat hearts.

OBJECTIVE:To optimize the extraction process for the preparation of Paeonia lactiflora formula granules and establish an HPLC method for the determination of Paeoniflorin in the samples.METHODS:The optimum water extraction process was selected by the L9(34)orthogonal design and the content of Paeoniflorin in formula granules was analyzed by HPLC.The HPLC assay was performed on a crosmosil C18-MS-II(250mm? 4.6mm,5? m)column.The mobile phase was employed by using acetonitrile-0.1% phosphoric acid(14∶ 86).The detection wavelength was set at 230 nm and the system was operated at 25℃.RESULTS:The optimum extraction conditions were to reflux for 3 times with 12 fold water and each time for one hour.The extraction rate of Paeoniflorin was 85.7% and the dried extraction yield was 34.4%.The peak area had a good linearity with the concentration of Paeoniflorin and the linear range was 0.128～ 0.640? g(r=0.999 8).The average recovery was 100.76% and RSD was 1.26%.CONCLUSION:The preparation process of Paeonia lactiflora formula granules was stable and feasible.The HPLC method for quality control was accurate and suitable.

OBJECTIVE:To develop HPLC method for quantitative determination of ti anepine sodium in blood plasma METHODS:Tianepine was extracted with ethyl acet ate The residues were analyzed with a reverse phase HPLC system(DiamonsilTM C18 column,4 6mm?250mm,5?m);Mobile phase,MeOH-C2H3OONH4(80∶20);UV detecti on 220nm RESUL_TS:The average recoveries for tianepine sodium in high,medium and low concentr ations were 97 63%,97 60% and 93 67%,respectively The within-day and b etween-day relative standard deviations were less than 5%(n=5) The calibratio n curves had good linearity within a concentration range of 1 0～83 3?g/ml T he regression equation was Y=6 05?10-2X+8 18?10-1,r=0 9 994(n=12) The limit of quantitation for tianepine sodium was 0 2?g/ml CONCLUSION:The metho d provides a se_nsitive,accurate,precise and reliable analytical procedure for clinical monito ring of tianepine sodium blood in plasma and its pharmacokinetic studies

Objective　To observe the effects of epithelial growth factor (EGF) and basic fibroblast growth factor (bFGF) on wound healing of the penetration keratoplasty (PKP) with autograft in rabbits. Methods　After the establishment of penetrating keratic autograft model on 24 rabbits, they were equally divided into EGF-, bFGF-, EGF+bFGF-treated and control groups. The wound healing of all the animals were observed with healing intensity, liquid scintillation counter, AgNORs staining, VG staining and transmission electron microscopy. Results　①EGF, bFGF, and EGF+bFGF increased the limiting pressure of the wounds and the 3H-TdR incorporation. ②The fibroblast cells and its secreting proliferative collagen in both the bFGF group and EGF+bFGF group were more well-arranged. Conclusion　①EGF, bFGF, EGF+bFGF can obviously elevate the intensity of wound healing after PKP and enhance the synthesis of DNA. ②The effect of this agent combination is just as the same as bFGF applied alone 14 d after the operation. ③bFGF can improve the quality of wound healing after PKP.

BACKGROUND: There are many methods to establish VX2 subcutaneous tumor model, and the existing modeling methods have their own advantages and disadvantages. To meet the needs of tumor experiments on animals, the method of establishing an animal tumor model with high-quality and high-efficiency is necessary. OBJECTIVE: To explore more simple and efficient way of modeling, provide rabbit VX2 subcutaneous tumor experiment with large quantities of high-quality animal models by comparing different methods to establish rabbit VX2 subcutaneous tumor model. METHODS: Sixty-six male New Zealand white rabbits were enrolled. Two of the 66 rabbits were used to prepare VX2 tumor-bearing rabbits, and tumor tissues were taken from the tumor-bearing rabbits to prepare tumor tissue blocks and tumor tissue suspensions. There were two groups in the experiment. In tumor tissue suspension group (n=20), the rabbits were injected with 0.15 mL of tissue suspension on the medial side of bilateral hind limbs after anesthesia; in tumor tissue block group (n=20), tumor tissue blocks were implanted subcutaneously on the medial side of bilateral hind limbs after anesthesia. Two tumor-bearing rabbits from each group were subjected to the corresponding vaccination methods, each passed for five generations. Tumor inoculation time in the two groups was record and compared. The tumor size and growth were observed by ultrasound with 2-D and CDFI mode. Tumor-take rate and serial passage of tumor tissues were observed and compared between the two groups. The experimental protocol was approved by the Animal Experiment Ethics Committee of the Army Medical University of PLA (approval No. AMUWE2020016) RESULTS AND CONCLUSION: The tumor inoculation time in the tissue suspension group [(75.70±11.16) s] was significant shortened compared with that in the tissue block group [(100.80±9.21) s; P=0.00]. The tumor-take rate was significantly higher in the tissue suspension group than the tissue block group (95% vs. 60%; P < 0.05). The tumor size was significantly larger in the tissue suspension group than the tissue block group (P < 0.05). The rate of tumor tissue series passage was significantly higher in the tissue suspension group than the tissue block group (95% vs. 65%; P < 0.05). Therefore, tissue suspension method for making the model of rabbit VX2 subcutaneous tumor is simpler and more efficient compared with the tissue block method.

Objective To identify the morphological parameters that are related to intracranial aneurysms (IAs) rupture using a case-control model. Methods A total of 107 patients with multiple IAs and aneurysmal subarachnoid hemorrhage between August 2011 and February 2017 were enrolled in this study. Characteristics of IAs location, shape, neck width, perpendicular height, depth, maximum size, flow angle, parent vessel diameter (PVD), aspect ratio (AR) and size ratio (SR) were evaluated using CT angiography. Multiple logistic regression analysis was used to identify the independent risk factors associated with IAs rupture. Receiver operating characteristic curve analysis was performed on the final model, and the optimal thresholds were obtained. Results IAs located in the internal carotid artery (ICA) was associated with a negative risk of rupture, whereas AR, SR1 (height/PVD) and SR2 (depth/PVD) were associated with increased risk of rupture. When SR was calculated differently, the odds ratio values of these factors were also different. The receiver operating characteristic curve showed that AR, SR1 and SR2 had cut-off values of 1.01, 1.48 and 1.40, respectively. SR3 (maximum size/PVD) was not associated with IAs rupture. Conclusions IAs located in the ICA are associated with a negative risk of rupture, while high AR (>1.01), SR1 (>1.48) or SR2 (>1.40) are risk factors for multiple IAs rupture.

AIMThe effects of angiotensin II on the changes of Ca2+ signal in cultured rat neonatal myocytes were investigated in order to reveal the localization and distribution of elementary Ca2+ signaling units.

METHODSThe cultured neonate rat myocytes were treated with angiotensin II, and calcium signal was detected using confocal laser scanning microscopy and fluo-4/AM calcium probe.

RESULTSThe propagation of Ca2+ waves was observed in rest and angiotensin II stimulated cardiac myocytes. Calcium fluorescent intensity oscillated slightly in myocytes and the average intensity was much higher in the nucleus than in the cytosol, all of which could be magnified significantly by AngII (10(-6) mol/L). Ca2+ oscillation induced by Ang II was completely blocked by NO donor sodium nitroprusside. AngII evoked Ca2+ sparks close to the cell surface membrane, and couldn't be abolished by sodium nitroprusside.

CONCLUSIONThere are spatiotemporal dynamics of Ca2+ signaling patterns such as Ca2+ wave, Ca2+ spikes, Ca2+ oscillation and the whole cell Ca2+ transients induced by angiotensin II, which might play very important roles in cellular cardiac function.

Angiotensin II ; pharmacology ; Animals ; Calcium ; metabolism ; Calcium Signaling ; Cells, Cultured ; Microscopy, Confocal ; Myocytes, Cardiac ; drug effects ; metabolism ; Rats