Novel Copper Complexes That Inhibit the Proteasome and Trigger Apoptosis in Triple-Negative Breast Cancer Cells.

Five innovative ternary copper(II) complexes [Cu(OH-PIP)(Phe)Cl](1), [Cu(OH-PIP)(Gly)(H2O)]NO3·2H2O (2), [Cu(OH-PIP)(Ala)(Cl)]·H2O (3), [Cu(OH-PIP)(Met)]PF6·2H2O (4), and [Cu(OH-PIP)(Gln)(H2O)](Cl)·3H2O (5) have been synthesized and characterized by infrared spectroscopy, elemental analysis, and single crystal X-ray diffraction analysis. X-ray crystallography indicates that all Cu atoms are five-coordinated in a square-pyramidal configuration. The complexes have been screened for cytotoxicity against human breast cancer cell lines MCF-7, MDA-MB-231, and CAL-51. The best anticancer activity is obtained with triple-negative breast cancer CAL-51 and MDA-MB-231 cell lines, with IC50 values in the range of 0.082-0.69 µM. Importantly, the copper compounds were more effective than carboplatin at triggering cell death. Mechanistically, the complexes inhibit proteasomal chymotrypsin-like activity, and docking studies reveal their 20S proteasome binding sites. As a consequence, they cause the accumulation of ubiquitinated proteins, inhibit cell proliferation, and induce apoptosis. In addition, these copper complexes decrease the stemness of triple-negative breast cancer cells and have synergistic effects with CBP on TNBC cells, indicating their great potential as a novel therapy for triple-negative breast cancer.

[Effects of nitrogen form on growth and quality of Chrysanthemums morifolium].

This paper is aimed to study the effects of nitrogen form on the growth and quality of Chrysanthemums morifolium at the same nitrogen level. In order to provide references for nutrition regulation of Ch. morifolium in field production, pot experiments were carried out in the greenhouse at experimental station of Nanjing Agricultural University. Five proportions of ammonium and nitrate nitrogen were set up and a randomized block design was applied four times repeatedly. The results showed that the growth and quality of Ch. morifolium were significantly influenced by the nitrogen form. The content of chlorophyll and photosynthesis rate were the highest at the NH4(+) -N /NO3(-) -N ratio of 25:75; The activities of NR in different parts of Ch. -morifolium reached the highest at the NH4(+) - N/NO3(-) -N ratio of 0: 100. The contents of nitrate nitrogen in the root and leaves reached the highest at the NH4(+) -N/NO3(-) -N ratio of 50:50. The activities of GS, GOGAT and the content of amylum increased with the ratio of NO3(-) -N decreasing and reached it's maximum at the NH4 + -N/NO3 - -N ratio of 100: 0. The content of ammonium nitrogen were the highest at the NH4 + -N /NO3 --N ratio of 75: 25, while the content of soluble sugar reached the highest at the NH4(+)-N/NO3(-) -N ratio of 25: 75. The content of flavones, chlorogenic acid and 3,5-O-dicoffeoylqunic acid were 57.2 mg x g(-1), 0.673% and 1.838% respectively, reaching the maximum at the NH4(+) -N /NO3(-) -N ratio of 25:75; The content of luteoloside increased with the ratio of NO3(-) -N increasing and reached it's maximum at the NH4(+) -N/NO3(-) -N ratio of 0: 100. The yield of Ch. morifolium reached it's maximum at the NH4(+) -N /NO3(-) -N ratio of 25:75. Nitrogen form has some remarkable influence on the nitrogen metabolism, photosynthesis and growth, Nitrogen form conducive to the growth and quality of Ch. morifolium at the NH4(+) -N /NO3(-) -N ratio of 25: 75.

[Effects of low-level laser irradiation on rat mesenteric microcirculatory disturbance during early stage of endotoxemia].

OBJECTIVE: To observe the effects of low-level laser irradiation on mesenteric microcirculation of rats in vivo in the early stage of endotoxemia (ETM). METHODS: The experimental model of ETM was reproduced by injection of lipopolysaccharide (LPS). Sixty healthy male Sprague-Dawley (SD) rats were divided into three groups used random number table: control group, LPS group and low-level laser irradiation group, each group included 20 rats which were subdivided into four temporal subgroups (1, 2, 4, 6 hours, respectively). In low-level laser irradiation group, the rats were irradiated by type SLT semiconductor laser (650 nm, 5 mW) on unilateral femoral artery and vein, and blood vessel of the ear concurrently for 30 minutes. The interference course was vertical irradiation taken at 30 minutes after the injection of LPS. At 1, 2, 4, 6 hours after the injection of LPS, changes in mesenteric microcirculation and microcirculatory blood flow were recorded with the laser Doppler flowmeter, the velocity of red blood cells in venules was observed, and the number of open capillaries and adherent leukocytes were recorded. RESULTS: The blood flow velocity (mm/s) of the mesenteric microcirculation in LPS group was accelerated at 1 hour and 2 hours after LPS injection (1 hour: 0.190+/-0.007 vs. 0.174+/-0.009, 2 hours: 0.200+/-0.010 vs. 0.172+/-0.015, both P<0.05, respectively), but decelerated at 6 hours (0.116+/-0.015 vs. 0.164+/-0.011, P<0.05). The blood flow volume in the mesenteric vessels and the number of open capillaries did not show any significant change at that time. Significant increase in number of adherent leukocytes was observed at 2, 4, 6 hours after injury (2 hours: 2.60+/-1.14 vs. 0.40+/-0.55, 4 hours: 5.40+/-0.89 vs. 0.40+/-0.55, 6 hours : 5.40+/-1.52 vs. 0.60+/-0.90, all P<0.05, respectively). The state of blood flow in the microcirculation became abnormal. After irradiated with laser in low dose, the blood flow velocity was smooth and stable (mm/s, 1 hour: 0.174+/-0.011, 2 hours: 0.180+/-0.023, 4 hours: 0.168+/-0.013, 6 hours: 0.162+/-0.023), and the number of adherent leukocytes was reduced significantly at 4 hours and 6 hours than that in LPS group (4 hours: 2.00+/-0.71 vs. 5.40+/-0.89, 6 hours: 2.60+/-1.52 vs. 5.40+/-1.52, both P<0.05) and the microcirculatory flow state was improved obviously. CONCLUSION: Low-level laser irradiation may ameliorate the local mesenteric microcirculation, alleviate the microcirculatory disorder in early stage of ETM.

A clinically relevant orthotopic xenograft model of ependymoma that maintains the genomic signature of the primary tumor and preserves cancer stem cells in vivo.

Limited availability of in vitro and in vivo model systems has hampered efforts to understand tumor biology and test novel therapies for ependymoma, the third most common malignant brain tumor that occurs in children. To develop clinically relevant animal models of ependymoma, we directly injected a fresh surgical specimen from a 9-year-old patient into the right cerebrum of RAG2/severe complex immune deficiency (SCID) mice. All five mice receiving the initial transplantation of the patient tumor developed intracerebral xenografts, which have since been serially subtransplanted in vivo in mouse brains for 4 generations and can be cryopreserved for long-term maintenance of tumorigenicity. The xenograft tumors shared nearly identical histopathological features with the original tumors, harbored 8 structural chromosomal abnormalities as detected with spectral karyotyping, maintained gene expression profiles resembling that of the original patient tumor with the preservation of multiple key genetic abnormalities commonly found in human ependymomas, and contained a small population (<2.2%) of CD133(+) stem cells that can form neurospheres and display multipotent capabilities in vitro. The permanent cell line (BXD-1425EPN), which was derived from a passage II xenograft tumor and has been passaged in vitro more than 70 times, expressed similar differentiation markers of the xenograft tumors, maintained identical chromosomal abnormalities, and formed tumors in the brains of SCID mice. In conclusion, direct injection of primary ependymoma tumor cells played an important role in the generation of a clinically relevant mouse model IC-1425EPN and a novel cell line, BXD-1425EPN. This cell line and model will facilitate the biological studies and preclinical drug screenings for pediatric ependymomas.

N-(3,4-Diethoxy-phen-yl)acetamide.

In the title compound, C(12)H(17)NO(3), the conformations of the N-H and C=O bonds are anti to each other. In the crystal structure, N-H⋯O hydrogen-bond inter-actions help to establish the packing.

[Effects of anisodamine on microcirculation of the asystole rats during the cardiopulmonary resuscitation].

OBJECTIVE: To study the effects of anisodamine (Ani) on microcirculation and reperfusion volume of intestine wall in Wistar rats during cardiopulmonary resuscitation (CPR) for cardiac arrest (CA). METHODS: Healthy Wistar rats subjected to CA and resuscitation were randomly divided into four groups, 15 rats in each group. After a 4-minute-non-intervention interval, CPR was started. After CPR for 4 minutes the animals in control group received normal saline, group epinephrine (Epi) received Epi (bolus dose of 200 microg/kg), Epi plus low dosage Ani (Ld Ani) group received Epi plus Ani (bolus dose of 200 microg/kg Epi followed by Ani of 5 mg/kg), and Epi plus high dosage Ani (Hd Ani) group received Epi plus Ani (bolus dose of 200 microg/kg Epi followed by Ani of 10 mg/kg). The recanalization rate of mesenteric arterioles and venules, caliber of the recanalized mesentery arteriole and venule, and the reperfusion volume of intestine wall were observed in vivo in rats with restoration of spontaneous circulation (ROSC). RESULTS: As the rate of recanalization of mesenteric arterioles and venules was compared, group Hd Ani (66.6%, 60.0%)>group Ld Ani (60.0%, 53.3%)>group control (40.0%, 40.0%)>group Epi (26.7%, 20.0%), and group Ld Ani and group Hd Ani was much better than group Epi(all P<0.05). When the caliber of arterioles and venules was compared, group Hd Ani>group Ld Ani>group control>group Epi 30 minutes and 60 minutes after ROSC. Thirty minutes after ROSC, the caliber of arterioles and venules was much larger in group Ld Ani and group Hd Ani than that in group Epi (all P<0.05). Sixty minutes after ROSC, there was no statistical difference in the caliber of venules between group Ld Ani and group Epi, so as between group Ld Ani and group Hd Ani. Reperfusion volume of intestine wall in group Ld Ani and group Hd Ani was higher than that in groups control and Epi 15 minutes after ROSC, and it kept on to be better up to 60 minutes after ROSC. CONCLUSION: Administration of Ani at earlier period of resuscitation could improve microcirculation of the tissue and raise ROSC rate and successful rate of resuscitation.

[Perlecan participates in the protection of ischemic myocardium induced by basic fibroblast growth factor: experiment with rats].

OBJECTIVE: To investigate if perlecan (PN) is involved in the myocardial protection of basic fibroblast growth factor (bFGF) in acute myocardial infarction. METHODS: Twenty-four Wistar rats were randomized into 3 groups: myocardial infarction group (MI group, n = 8, undergoing ligation of the descending anterior branch of the left coronary artery), bFGF + MI group (n = 10, injected with bFGF into the border myocardium between the infracted and non- infracted areas immediately after the ligation of the descending anterior branch), and sham operation group (n = 6, undergoing sham operation and injection of normal saline). 24 h, 14 days, and 28 days after the operation the hemodynamic parameters, infarct size, and microvessel density (MVD) were observed. The hearts were taken out, RT-PCR was used to detect the mRNA expression of PN, and Western blotting was used to detect the expression of focal adhesion kinase (FAK) and p38 mitogen-activated protein kinase (MAPK) phosphorylation. RESULTS: In the bFGF + MI group, the +dp/dt and -dp/dt were improved markedly, the infarct size was significantly smaller, the MVD value was higher than those of the MI group on day 14 and day 28; the perlecan mRNA expression was higher in the infarct marginal area and interior zone; and the expression of FAK and the p38MAPK phosphorylation were up-regulated in the marginal zone of the bFGF group. CONCLUSION: bFGF is useful in promoting ischemic myocardial angiogenesis; reducing the size of infracted myocardium, and improving the ventricular function in acute myocardial infarction. Perlecan participates in the cardiac protection induced by bFGF. The relevant pathway is related to up-regulation of FAK and p38MAPK.

[Histopathology analysis of microemboli after thrombolysis].

OBJECTIVE: To observe the changes in fibrin of microemboli in micro-embolism soon after thrombolysis, in order to provide a theoretical basis for micro-thrombolytic therapy. METHODS: Thirty adult male SD rats weighing (180+/-10)g were randomized into five groups: 5, 30, 60 and 90 minutes groups after micro-thromboembolization and the control group. Rat cremasteric artery was embolized with photochemical method, and micro-thrombi were produced after thrombolysis by staphylokinase. The levels of tissue plasminogen activator (t-PA) and tissue plasminogen activator inhibitor (PAI) were immunohistochemically determined, and taken as the markers of fibrin dissolution. RESULTS: Fibrin was observed in pathologic sections as well as fibrinolytic markers t-PA and PAI in immunohistochemical sections. The levels of t-PA were found to be reduced while that of PAI increased gradually with passage of time. CONCLUSION: Fibrin is one of the main constituents of microthrombi soon after thrombolysis. The content of fibrous emboli increases after microembolization with the passage of time. Thrombolysis plays an important role in the treatment of "no reflow phenomenon".

[The changes in effective local blood perfusion and compensatory ventilation in different lung areas of acute respiratory distress syndrome rabbits model].

OBJECTIVE: To discuss the mechanism of severe hypoxemia that induced with acute respiratory distress syndrome (ARDS) by way of observing the effective local blood perfusion in different lung areas of ARDS rabbits. METHODS: ARDS rabbit models were established by injecting oleic acid through central vein, and the results of effective local blood perfusion in different areas of rabbit lung (upper area, abdominal area, and dorsal area of right lung)with PIM-II laser perfusion imager and arterial blood gas under different ventilation modes [large tidal volume ventilation, low tidal volume ventilation + positive end-expiratory pressure (PEEP), prone position + large tidal volume ventilation, and prone position + low tidal volume ventilation + PEEP] were measured. RESULTS: The results of oxygenation index were significantly getting worsen after lung injury, and after receiving lung protective ventilation (including low tidal volume + PEEP, prone position + low tidal volume + PEEP), the indexes were taking a turn to the better. After lung injury, the results of effective local lung blood perfusion index (including upper area, abdominal area, and dorsal area) under right lung of ARDS rabbits decreased in the experimental ARDS groups. The results of experimental dorsal area group decreased most significantly, and the results of experimental upper area group changed most lightly. We observed that the effects of receiving low tidal volume + PEEP ventilation were not as good as prone position + low tidal volume + PEEP ventilation in the dorsal area of the rabbit lung compared to the results of effective local area lung blood perfusion index between two ventilation modes, there showed significantly differences in statistics. CONCLUSION: We indicated that the one of the main mechanisms of severe hypoxemia in ARDS might be caught by the shunt between pulmonary artery and pulmonary vein just like physiological right to left shunt, which caused severe ventilation/perfusion disturbance. Both low tidal volume ventilation + PEEP and prone position + low tidal volume ventilation + PEEP had good effects on ameliorating local blood perfusion, and the effects of ventilation mode of prone position + low tidal volume ventilation +PEEP might be better.