Paeoniflorin Enhances the Sensitivity of ER-Positive Breast Cancer Cells to Tamoxifen through Promoting Sirtuin 4.

Tamoxifen is an effective drug for treating patients with advanced estrogen receptor-positive (ER+) breast cancer (BC), but not for all ER + BC patients. Drug tolerance is the biggest obstacle. In this study, we designed an experiment to investigate whether paeoniflorin affects the ER + BC cell's sensitivity to tamoxifen in the T47D and MCF-7 cell lines. Herein, we found that paeoniflorin inhibited cell proliferation without inducing apoptosis. However, it enhanced tamoxifen-induced apoptosis in both cell lines. Immunoblotting revealed that paeoniflorin significantly increased the already elevated Bax/Bcl2 protein expression ratio and the caspase 3 activity levels, both induced by tamoxifen. Paeoniflorin was also found to increase SIRT4 expression, and deletion of SIRT4 could significantly reverse the inhibition of cell proliferation induced by paeoniflorin and significantly decrease paeoniflorin-enhanced apoptosis induced by tamoxifen. Moreover, protein expression detection revealed that paeoniflorin enhanced the tamoxifen-induced inhibition of STAT3 activation. Besides, the deletion of SIRT4 could significantly increase STAT3 activation in the T47D and MCF-7 cells. In conclusion, paeoniflorin suppressed STAT3 activation to enhance the sensitivity of ER-positive breast cancer cells to tamoxifen through promoting SIRT4 expression.

Evaluation of macular visual function in patients with myopic choroidal neovascularization before and after intravitreal injection of conbercept / 中华眼底病杂志

Objective To evaluate the macular visual function of patients with myopic choroidal neovascularization (MCNV) before and after intravitreal injection of conbercept.Methods A prospective,uncontrolled and non-randomized study.From April 2017 to April 2018,21 eyes of 21 patients diagnosed as MCNV in Shanxi Eye Hospital and treated with intravitreal injection of conbercept were included in this study.There were 9 males (9 eyes,42.86％) and 12 females (12 eyes,57.14％),with the mean age of 35.1 ± 13.2 years.The mean diopter was-11.30 ± 2.35 D and the mean axial length was 28.93 ± 5.68 mm.All patients were treated with intravitreal injection of conbercept 0.05 ml (1+PRN).Regular follow-up was performed before and after treatment,and BCVA and MAIA micro-field examination were performed at each follow-up.BCVA,macular integrity index (MI),mean sensitivity (MS) and fixation status changes before and after treatment were comparatively analyzed.The fixation status was divided into three types:stable fixation,relatively unstable fixation,and unstable fixation.The paired-sample t-test was used to compare BCVA,MI and MS before and after treatment.The x2 test was used to compare the fixation status before and after treatment.Results During the observation period,the average number of injections was 3.5.The logMAR BCVA of the eyes before treatment and at 1,3,and 6 months after treatment were 0.87±0.32,0.68±0.23,0.52±0.17,and 0.61 ±0.57,respectively;MI were 89.38 ± 21.34,88.87 ± 17.91,70.59 ± 30.02,and 86.76 ± 15.09,respectively;MS were 15.32 ± 7.19,21.35 ± 8.89,23.98 ± 11.12,22.32 ± 9.04 dB,respectively.Compared with before treatment,BCVA (t=15.32,18.65,17.38;P＜0.01) and MS (t=4.08,3.50,4.26;P＜0.01) were significantly increased in the eyes 1,3,and 6 months after treatment.There was no significant difference in the MI of the eyes before treatment and at 1,3,and 6 months after treatment (t=0.60,2.42,2.58;P＞0.05).Before treatment and at 1,3,and 6 months after treatment,the proportion of stable fixation were 28.57％,38.10％,38.10％,33.33％;the proportion of relatively unstable fixation were 47.62％,47.62％,52.38％,57.14％ and the proportion of unstable fixation were 23.81％,14.28％,9.52％,9.52％,respectively.The proportion of stable fixation and relatively unstable fixation at 1,3 and 6 months after treatment were higher than that before treatment,but the difference was not statistically significant (x2=1.82,1.24,1.69;P＞0.05).Conclusion BCVA and MS are significantly increased in patients with MCNV after intravitreal injection of conbercept.

Study of forced degradation behavior of pramlintide acetate by HPLC and LC-MS.

Pramlintide acetate (Symlin®), a synthetic analogue of the human hormone amylin. It was approved in March 2005 as a subcutaneous injection for the adjunctive treatment of patients who have type 1 or 2 diabetes mellitus. The objective of current investigation was to study the degradation behavior of pramlintide acetate under different ICH recommended stress conditions by HPLC and LC-MS. Pramlintide acetate was subjected to stress conditions of hydrolysis (acidic or alkaline), oxidation, photolysis and thermal decomposition. Extensive degradation products were observed under the hydrolysis, oxidation or thermal stress conditions, while minimal degradation was found in the photolytic conditions. Successful separation of drug from the degradation products was achieved by the validated chromatography (RP-HPLC and SCX-HPLC) methods. Subsequent to isolation, the molecular weight of each component was determined by LC-MS. The LC-MS m/z values and fragmentation patterns of 4 impurities matched with the predicted degradation products of pramlintide acetate.

Mechanism of glaucocalyxin A that induces apoptosis of glioma cell and up-regulates GEF-H1 / 药学学报

Rabdosia japonica(Burm.f.) Hara var.glaucocalyx(Maxim.) Hara is a traditional Chinese medicine, and is known to have anti-tumor effects. This study aims to investigate the effect of glaucocalyxin A (GLA), a diterpenoids extracted from Glaucocalyx Hara, on apoptosis of glioma cells and its mechanism. This study investigated the molecular signaling mechanism of GLA-induced glioma cell apoptosis by analyzing survival rate of C6 rat glioma cells, cell morphology, colony formation ability, interference ribonucleic acid, polymerase chain reaction, and Western blot. The result showed that in the presentce of GLA, the survival rate of C6 rat glioma cells decreased significantly, while the expression of guanine nucleotide-exchange factor-H1 was up-regulated, causing phosphorylation of extracellular regulated protein kinases proteins and apoptosis. Hence, the mechanism of GLA-induced glioma cell apoptosis was the GEF-H1/ERK pathway.

[Quantitative assess the efficacy of congenital idiopathic nystagmus surgery by digital eye tracker].

OBJECTIVE: To evaluate the feasibility of independently developed digital eye tracker in determining the efficacy of congenital idiopathic nystagmus (CIN) surgery. METHODS: Aprospective selfpairing study. The surgical efficacy was evaluated by independently developed eye tracker in 16 CIN patients. The null zone and the frequency, amplitude, intensity of nystagmus in various gazing position were recorded with eye tracker pre and post operatively. The consistency of null zone determined by digital eye tracker and clinical investigation were evaluated. The preoperative and postoperative rectification of horizontal and vertical nystagmus in the horizontal direction of individual patient were compared by paired samples t-test. The improvement or aggravation quantity were recorded by comparing preoperative and postoperative intensity of nystagmus every 5° within 25°. RESULT: The null zone got from digital eye tracker and clinical investigation were highly consistent (r = 0.952, P < 0.01). The horizontal and vertical intensity improved in 9 patients (t = 2.335-6.609, P < 0.05) and 5 patients (t = 2.176-5.471, P < 0.05) respectively after surgery. There were 67.63% (117/173) horizontal intensity and 69.94% (121/173) vertical intensity improvement. CONCLUSION: The independently developed digital eye tracker can quantitatively evaluate the pre and post-operative nystagmus and analyze the surgical efficacy for CIN patients.

Effect of artificial scotomas on open-loop disparity vergence eye movements.

PURPOSE: To investigate the effect of an artificial scotoma on open-loop disparity vergence responses (DVRs) and vergence control mechanisms, we examined open-loop DVRs to disparity stimuli using monocular artificial scotomas in normal subjects. METHODS: Using a mirror haploscope with two computer monitors, we delivered disparity stimuli on a pair of random dot patterns subtending 40 by 30 degrees at 47 cm from each eye. The scotomas were black circles located in the center of a random dot pattern for the left eye. Eye movements of both eyes were recorded with a magnetic search coil system. RESULTS: We first found that the amplitudes of DVRs were gradually decreased and the latency of DVRs was moderately increased as the size of the scotomas was increased. Second, monocular responses from each eye were symmetrical although the stimuli to each eye were asymmetrical. CONCLUSIONS: The results suggest that the monocular eye movements in disparity vergence are controlled by a binocular central mechanism, not driven separately by monocular inputs in the open-loop window.

A new measure of nystagmus acuity.

AIM: To construct a new visual acuity measuring function for congenital nystagmus (CN) patients by studying the relationships between acuity, velocities and positions of the eye. METHODS: After assessing the relationship between acuity, movement velocities and positions of the eye separately, a new function, which we call the automated nystagmus acuity function (ANAF), was constructed to measure the visual acuity of CN patients. Using a high-speed digital video system working at 500 frames per second, each eye was calibrated during monocular fixation. Twenty-six recorded nystagmus data were selected randomly. Using nystagmus waveforms, the best vision position (foveation period) and visual acuity were analyzed in three groups of subjects, and then all outputs were compared with the well-known expanded nystagmus acuity function (NAFX) and ANAF. Standard descriptive statistics were used to summarize the outputs of the two programs. RESULTS: Foveation periods were brief intervals in the CN waveform when the image was on or near the fovea and eye velocity was relatively slow. Results showed good visual acuity happened during the period when velocity was low and the eye position was near the zero position, which fitted the foveation periods. The data analyzed with NAFX and ANAF had a correlation coefficient of 0.934276, with an average error of -0.00973. CONCLUSION: The results from ANAF and NAFX analyses showed no significant difference. The NAFX manually identifies foveation eye positions and produces accurate measurements. The ANAF, however, can be calculated simply using the factors eye position and velocity, and it automatically calculates the ANAF without the need to manually identify foveation eye positions.

Identification of small molecule activators of the janus kinase/signal transducer and activator of transcription pathway using a cell-based screen.

Type I interferons (IFN-α/ß) have been widely used in the treatment of many viral and malignant diseases by activation of the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway, but the side effects of protein-based IFN therapy severely limit their clinical use. Discovering small molecules to activate the JAK/STAT pathway will greatly facilitate the development of new drugs which have similar pharmacological function to IFNs but with fewer side effects. To screen a natural products-based library, we established a cell-based screening assay using human hepatoma HepG2 cells stably transfected with a plasmid where the luciferase reporter activity is driven by interferon α-stimulated response element (ISRE), the motif specifically recognized by type I IFN-induced activation of JAK/STAT pathway. Among 1,431 natural product compounds screened, four compounds (emodin, quercetin, apigenin and luteolin) were identified as activators of the JAK/STAT pathway. Further studies demonstrated that these four compounds could increase the endogenous antiviral gene expression regulated by the IFN-activated JAK/STAT pathway. The identified small molecule activators are valuable for structural modification and warrant further investigation for use in new antiviral drugs as IFN mimics or adjuvants.

MyMolDB: a micromolecular database solution with open source and free components.

BACKGROUND: To manage chemical structures in small laboratories is one of the important daily tasks. Few solutions are available on the internet, and most of them are closed source applications. The open-source applications typically have limited capability and basic cheminformatics functionalities. In this article, we describe an open-source solution to manage chemicals in research groups based on open source and free components. It has a user-friendly interface with the functions of chemical handling and intensive searching. RESULTS: MyMolDB is a micromolecular database solution that supports exact, substructure, similarity, and combined searching. This solution is mainly implemented using scripting language Python with a web-based interface for compound management and searching. Almost all the searches are in essence done with pure SQL on the database by using the high performance of the database engine. Thus, impressive searching speed has been archived in large data sets for no external Central Processing Unit (CPU) consuming languages were involved in the key procedure of the searching. AVAILABILITY: MyMolDB is an open-source software and can be modified and/or redistributed under GNU General Public License version 3 published by the Free Software Foundation (Free Software Foundation Inc. The GNU General Public License, Version 3, 2007. Available at: http://www.gnu.org/licenses/gpl.html). The software itself can be found at http://code.google.com/p/mymoldb/.

A new algorithm for automated nystagmus acuity function analysis.

AIMS: We developed a new data analysis algorithm called the automated nystagmus acuity function (ANAF) to automatically assess nystagmus acuity function. We compared results from the ANAF with those of the well-known expanded nystagmus acuity function (NAFX). METHODS: Using the ANAF and NAFX, we analysed 60 segments of nystagmus data collected with a video-based eye tracking system (EyeLink 1000) from 30 patients with infantile or mal-development fusional nystagmus. The ANAF algorithm used the best-foveation positions (not true foveation positions) and all data points in each nystagmus cycle to calculate a nystagmus acuity function. RESULTS: The ANAF automatically produced a nystagmus acuity function in a few seconds because manual identification of foveation eye positions is not required. A structural equation model was used to compare the ANAF and NAFX. Both ANAF and NAFX have similar measurement imprecision and relatively little bias. The estimated bias was not statistically significant for either methods or replicates. CONCLUSIONS: We conclude that the ANAF is a valid and efficient algorithm for determining a nystagmus acuity function.

A systematic approach to eye muscle surgery for infantile nystagmus syndrome: results in 100 patients.

PURPOSE: To report a systematic approach to, and the visual and electrophysiological effect of, eye muscle surgery in 100 patients with infantile nyustagmus syndrome (INS). METHODS: Prospective, interventional case cohort analysis of clinical and eye movement data in 100 patients with INS who had virgin extraocular eye muscles operated on for nystagmus with or without combinations of strabismus and an anomalous head posture. All patients were followed at least 9 months after surgery. Outcome measures, part of an IRB approved study, included binocular visual acuity, head position, strabismic deviation, and eye movement recordings, from which waveform types and an Automated Nystagmus Acuityn Function (ANAF) was calculated. Computerized parametric and non-parametric statistical analysis of data were performed using standard software on both individual and group data. RESULTS: There were 9 consistent surgical procedures used with the most common being that for a horizontal head posture alone (22%). Age at surgery averaged 14 years with 11 months followup. Sixty-eight percent had associated eye disease (optic nerve, retinal, amblyopia, cataracts). Group means in binocular acuity, strabismic deviation, head posture, abd ANAF measures from eye improved for all procedures. There were 12 (12%) reoperations without any serious surgical complications. Individual analysis revealed only age and head posture differences in outcome measures between the 9 procedures. CONCLUSIONS: Using this approach, surgery on the extraocular muscles in patients with INS results in improvements in multiple aspects of ocular motor and visual function.

A new hetisine-type alkaloid from the stems and leaves of Aconitum coreanum / 药学学报

<p><b>AIM</b>To study the chemical constituents of the stems and leaves of Aconitum coreanum (Lèvl.) Rapaics.</p><p><b>METHODS</b>The constituents of Aconitum coreanum were isolated by using various kinds of modern chromatographic methods. The new alkaloid was identified on the basis of spectral analysis.</p><p><b>RESULTS</b>Two compounds were isolated and identified as: 13-dehydro-1beta-acetyl-2alpha,6beta-dihydroxyhetisine (I) and Guanfu base G (II).</p><p><b>CONCLUSION</b>Compound I is a new alkaloid.</p>

Study on selective isolation of volatile oil in the seed of Fructus foeniculi / 中国中药杂志

<p><b>OBJECTIVE</b>To study the selective isolation of a single chemical component from volatile oil of Fructus foeniculi by inclucion method.</p><p><b>METHOD</b>The host molecule was selected and a single chemical component isolated from volatile oil by the host-guest recognition.</p><p><b>RESULT</b>X-ray single crystal analysis showed that 1,1,6,6-tetraphenylhexa-2, 4-diyne-1, 6-diol could successfully include 4-[1-propenyl] benzaldehyde from volatile oil of Fructus foeniculi.</p><p><b>CONCLUSION</b>The host-guest inclusion technology can be used to isolate a single component selectively from mixture.</p>

Establishment of drug screening model based on transcriptional regulation of estrogen responsive element / 中国中药杂志

<p><b>OBJECTIVE</b>AIM To establish a drug screening model based on transcriptional regulation of estrogen responsive element (ERE) and use it to screen compounds for discovering new ligands of estrogen receptor (ER) subtypes.</p><p><b>METHOD</b>A recombinant reporter vector pERE-TAL-SEAP was constructed by inserting a synthetic sequence composed of five tandem copies of EREs upstream of promoter of the reporter vector pTAL-SEAP. The pERE-TAL-SEAP and the internal control plasmid pCMV were transiently co-transfected into Hela cells expressing ER subtype or ER subtype, and the effects of pure ER agonists 17estradiol, phytoestrogen genistein and pure ER antagonist ICI182, 780 on reporter gene SEAP expression were observed.</p><p><b>RESULT</b>In the Hela cells expressing ER alpha or ER beta subtype, the expression of SEAP gene were induced in a dose dependent manner by 17-estrodiol with a maximal effect at approximately 10 nmol.L-1 and with EC50 of (80.58 +/- 8.51) pmol.L-1 and (103.90 +/- 5.29) pmol.L-1, respectively, so done by phytoestrogen genistein with a maximal effect at 1 mumol.L-1 and with EC50 of (10.86 +/- 0.75) nmol.L-1 and (39.38 +/- 2.26) nmol.L-1, respectively. The maximal level induced by estrodiol and genistein were about 7-14 fold higher than that of vehicle. The pure antiestrogen ICI182, 780 at concentration of 1 mumol.L-1 completely blocked the inductions of 17-estrodiol and genistein.</p><p><b>CONCLUSION</b>The cellular drug screening model can be established by transfecting reporter vector pERE-TAL-SEAP in Hela cell lines expressing ER alpha or ER beta. The cell lines can be used to screen compounds with estrogenicity by testing SEAP activity in the culture media of cells growing in microtitier wells. The system should provide an efficient model for screening and analyzing the activity of large numbers of ligands of ER.</p>