Effect of microRNA-221 targeting tissue inhibitor of metalloproteinase-2 to mediate Akt / mTOR signaling pathway on non-small cell lung cancer transplanted tumor mouse model / 解剖学报

Objective To explore the effects of miR-221 on tumor cell proliferation, migration and invasion in nonsmall cell lung cancer (NSCLC) xenograft model mice, and to preliminarily analyze its possible mechanism of regulating Akt/ mammalian target of rapamycin(mTOR) signaling pathway by targeting tissue inhibitor of metalloproteinase-2 (TIMP-2) on tumor cells in non-small cell lung cancer (NSCLC) through tumor-bearing nude mice. Methods The A549 cells were divided into control group, mimic group, TIMP-2 group and mimic+TIMP-2 group. The mimic group and TIMP-2 group were transfected with miR-221 mimic and TIMP-2 overexpression plasmids, respectively. The mimic + TIMP-2 group was simultaneously transfected with miR-221 mimic and TIMP-2 overexpression plasmids. The control group was transfected with the same amount of negative control plasmid. After transfection, the cells of each group were injected subcutaneously into the left forelimb to construct the corresponding 4 groups of NSCLC mouse models. The proliferation-related protein (Ki67) was detected by immunohistochemical staining to detected the effect of cell proliferation ability. Matrix metalloproteinase-2 (MMP-2) and N-cadherin proteins in each group were tested by Western blotting to assess and compare the abilities of migration and invasion. The levels of miR-221, TIMP-2 and Akt/ mTOR pathways in bone marrow and tumor tissues were detected by Real-time PCR and Western blotting. Results When co-transfected with wild type(WT)-TIMP-2 and miR-221 mimic, the relative luciferase activity in the cells reduced significantly (P<0. 05). Compared with the control group, the tumor mass, volume, Ki67, MMP-2 and N-cadherin protein expression levels, miR-221 and Akt/ mTOR pathway levels were increased significantly, while the levels of TIMP-2 mRNA and protein were significantly reduced in the mimic group (P<0. 05). Compared with the control group, the levels of TIMP-2 mRNA and protein in the TIMP-2 group increased significantly, while the other indicators decreased significantly (P<0. 05). Tumor tissue mass, volume, Ki67, MMP-2, Ncadherin, miR-221 and Akt/ mTOR pathway levels in mimic+TIMP-2 group were significantly lower than those in the mimic group and significantly higher than those in the TIMP-2 group, while TIMP-2 mRNA and protein levels were significantly higher than those in the mimic group and significantly lower than those in the TIMP-2 group (P<0. 05). Conclusion In the NSCLC transplanted tumor mouse model, miR-221 may mediate the Akt/ mTOR pathway by targeting the expression of TIMP-2 protein to promote cell proliferation, migration and invasion.

UPLC Fingerprint Analysis of Acanthopanacix Cortex from Different Origins / 中国实验方剂学杂志

Objective::To established fingerprint of Acanthopanacix Cortex by UPLC method, in order to provide reference for quality control and evaluation. Method::UPLC method was performed on Waters BEH C18 (2.1 mm×100 mm, 1.7 μm), with acetonitrile-0.1% glacial acetic acid as the mobile phase for gradient elution.The detection wavelength was 282 nm, the flow rate was 0.3 mL·min-1, the column temperature was 25 ℃, and the injection volume was 2 μL.With syringin as reference substance, the fingerprint of 20 batches Acanthopanacix Cortex were analyzed under the same chromatographic conditions.The Similarity Evaluation System for Chromatographic Fingerprint of Chinese Materia Media (version 2012) was used to analyze the similarity of 20 batches of Acanthopanacix Cortex, and the SPSS 21.0 was applied for cluster analysis. Result::The UPLC fingerprint of the Acanthopanacix Cortex was established.The similarity results showed that the 7 batches of the 20 batches of Acanthopanacix Cortex was less than 0.800, and the remaining medicinal materials were similar within the range from 0.800 to 0.924.Besides, 12 common fingerprint peaks were calibrated and 4 components were identified, namely protocatechuic acid (peak 1), chlorogenic acid (peak 3), syringin (peak 4), and 4-methoxysalicylaldehyde (peak 12). The clustering results showed that the 20 batches of Acanthopanacix Cortex were divided into four groups.Among these batches, S1, S3, S9, S13 and S20 were clustered into one category, S11 was a category, S14 was a category, and the remaining samples belonged to a category. Conclusion::With a good precision, repeatability and stability, short analysis time as well as superior specificity, the method will provide a scientific basis to evaluate and control the quality of Acanthopanacix Cortex.

Abnormal function of the posterior cingulate cortex in heroin addicted users during resting-state and drug-cue stimulation task / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Previous animal and neuroimaging studies have demonstrated that brain function in heroin addicted users is impaired. However, the posterior cingulate cortex (PCC) has not received much attention. The purpose of this study was to investigate whether chronic heroin use is associated with craving-related changes in the functional connectivity of the PCC of heroin addicted users.</p><p><b>METHODS</b>Fourteen male adult chronic heroin users and fifteen age and gender-matched healthy subjects participated in the present study. The participants underwent a resting-state functional magnetic resonance imaging (fMRI) scan and a cue-induced craving task fMRI scan. The activated PCC was identified in the cue-induced craving task by means of a group contrast test. Functional connectivity was analyzed based on resting-state fMRI data in order to determine the correlation between brain regions. The relationship between the connectivity of specific regions and heroin dependence was investigated.</p><p><b>RESULTS</b>The activation of PCC, bilateral anterior cingulate cortex, caudate, putamen, precuneus, and thalamus was significant in the heroin group compared to the healthy group in the cue-induced craving task. The detectable functional connectivity of the heroin users was stronger between the PCC and bilateral insula, bilateral dorsal striatum, right inferior parietal lobule (IPL) and right supramarginal gyrus (P < 0.001) compared to that of the healthy subjects in the resting-state data analysis. The strength of the functional connectivity, both for the PCC-insula (r = 0.60, P < 0.05) and for PCC-striatum (r = 0.58, P < 0.05), was positively correlated with the duration of heroin use.</p><p><b>CONCLUSION</b>The altered functional connectivity patterns in the PCC-insula and PCC-striatum areas may be regarded as biomarkers of brain damage severity in chronic heroin users.</p>

Effect of locally administered rapamycin and cyclosporin A on penetrating keratoplasty rejection rat model / 国际眼科杂志(Guoji Yanke Zazhi)

· AIM: To evaluate the combined effect of rapamycin ophthalmic solution and cyclosporin A ophthalmic solution on allogeneic transplantation in a rat model.rats were used as donors. The animals were randomly assigned to 4 groups. Group A: control (ophthalmic solution solvent 100μ L); group B: 2g/L rapamycin ophthalmic solution 100μ L; group C: 10g/L cyclosporin A ophthalmic solution 100μ L; group D: 2g/L rapamycin ophthalmic solution 50μ L and 10g/L cyclosporin A ophthalmic solution 50μ L, 4 times every day. The treatment was started on 2d after operation, and the animals were administered until rejection. The grafts were inspected by slit-lamp microscope and the corneal survival time was recorded. The pathologic changes were measured by light microscope.significantly prolonged compared with the control group (P ＜ 0.01). However, the combined therapy (group D) was significantly superior compared with group B and C (P＜0.05, P＜ 0.01). The histopathological findings showed that the inflammation cells, neovascularity in each treated group were significantly fewer than that in control group at 14d after operation.a double drug regimen with rapamycin ophthalmic solution and cyclosporin A ophthalmic solution for the control of acute corneal allograft rejection. It indicats that the combined therapy produced synergistic effect.

Determination of adefovir in monkey plasma by liquid chromatography-tandem mass spectrometry / 药学学报

<p><b>AIM</b>To develop a sensitive and specific liquid chromatography-tandem mass spectrometry (LC/MS/MS) method for the determination of adefovir in monkey plasma.</p><p><b>METHODS</b>Adefovir and internal standard 9-(3-phosphonylmethoxypropyl) adenine were isolated from plasma by protein precipitation with methanol, then chromatographed by using a Diamonsil C18 column. The mobile phase consisted of methanol-water-formic acid (20:80:1). Electrospray ionization (ESI) source was applied and operated in the positive ion mode. Selected reaction monitoring (SRM) mode with the transitions of m/z 274-->m/z 162 and m/z 288-->m/z 176 were used to quantify adefovir and the internal standard, respectively.</p><p><b>RESULTS</b>The linear calibration curve was obtained in the concentration range of 0.02-4.00 mg.L-1. The lower limit of quantitation was 20 micrograms.L-1. The inter- and intra-day precision (RSD) was less than 5.8%, and the accuracy (relative error) was within +/- 4.5%. The method was successfully used in a pharmacokinetic study of adefovir dipivoxil in monkeys.</p><p><b>CONCLUSION</b>The method is proved to be suitable for pre-clinical investigation of adefovir dipivoxil pharmacokinetics, which offers advantages of specificity and simple sample preparation compared with the previously reported methods.</p>