PF-06409577 Activates AMPK Signaling and Inhibits Osteosarcoma Cell Growth.

Osteosarcoma (OS) is a common primary bone malignancy. We here investigated the potential activity of PF-06409577, a novel, potent, and direct activator of AMP-activated protein kinase (AMPK), against human OS cells. In established (U2OS, MG-63, and SaOs-2 lines) and primary human OS cells, PF-06409577 inhibited cell viability and proliferation, while inducing cell apoptosis and cell cycle arrest. PF-06409577 induced AMPK activation, mTORC1 inhibition, autophagy induction, and downregulation of multiple receptor tyrosine kinase inOS cells. AMPK inactivation by AMPKα1 shRNA, CRISPR/Cas9 knockout, or dominant negative mutation (T172A) was able to abolish PF-06409577-induced activity in OS cells. In vivo, PF-06409577 oral administration at well-tolerated doses potently inhibited growth of U2OS cells and primary human OS cells in severe combined immunodeficient mice. AMPK activation, mTORC1 inhibition, autophagy induction, as well as RTK degradation and apoptosis activation were detected in PF-06409577-treated xenografts. In conclusion, activation of AMPK by PF-06409577 inhibits OS cell growth.

[Sampling time of living leaf for estimating phytolith-occluded organic carbon sequestration rate of Phyllostachys edulis.] / æ¯›ç«¹æž—æ¤ç¡ ä½“ç¢³å°å­˜é€ŸçŽ‡ä¼°ç®—çš„æœ€ä½³é²œå¶é‡‡æ ·æ—¶é—´.

Monthly phytolith-occluded organic carbon (PhytOC) content in living leaves and litterfall of Moso bamboo (Phyllostachys edulis) were measured for a year. The PhytOC sequestration rate in living leaves of different months were compared with that in annual litterfall to determine the optimum sampling time of living leaves for estimating PhytOC sequestration rate of Moso bamboo. The contents of phytoliths and PhytOC in living leaves of Moso bamboos were 23.45-101.07 g·kg-1 and 0.73-1.98 g·kg-1, respectively, with significant difference among different months. The monthly PhytOC sequestration rates of living leaves of Moso bamboo in different months ranged from 0.75 to 7.68 kg·hm-2·a-1. The maximum and minimum rates of the PhytOC sequestration occurred in December and April respectively, with significant difference between them. There was no difference between the PhytOC sequestration rate in living leaves of Moso bamboos in February or December and that of litterfall in the whole year. Therefore, February or December should be the optimal month of sampling living leaves for estimating the PhytOC sequestration rate of Moso bamboo stands.

The anti-cancer activity of the mTORC1/2 dual inhibitor XL388 in preclinical osteosarcoma models.

In the present study, we investigated the activity of XL388, a novel mammalian target of rapamycin (mTOR) complex 1/2 (mTORC1/2) dual inhibitor, in preclinical osteosarcoma (OS) models. XL388 was cytotoxic, cytostatic and pro-apoptotic to multiple established OS cell lines and primary human OS cells. XL388 blocked mTORC1/2 activation and downregulated cyclin D1/B1 expressions in OS cells, leaving AKT Thr-308 phosphorylation un-affected. Intriguingly, AKT1 T308A mutation potentiated XL388-induced cytotoxicity in OS cells. XL388 activated cytoprotective autophagy in OS cells. Autophagy inhibition, either pharmacologically or genetically, augmented XL388-induced anti-OS activity. Further, XL388 oral administration inhibited U2OS xenografts growth in severe combined immuno-deficient (SCID) mice. Such activity was enhanced with co-administration of the autophagy inhibitor 3-methyladenine (3-MA). Similarly, Beclin-1-silenced U2OS xenografts were remarkably more sensitive to XL388. Thus, concurrent blockage of mTORC1/2 with XL388 may have therapeutic value for OS.

[Numerical analysis of inter-specific relationships in the estuary salt marsh plant community of southern Chongming Dongtan, Shanghai.] / å´‡æ˜Žä¸œæ»©å—éƒ¨æ²³å£ç›æ²¼æ¤ç‰©ç¾¤è½ç§é—´å ³ç³»çš„æ•°é‡åˆ†æž.

The salt marsh plant communities were investigated with quadrats in the southern Chongming Dongtan. Based on the vegetation coverage and the 2×2 contingency table, 8 common species among the 17 higher plants recorded were analyzed. The variance ratio of overall association, Chi-square test and Spearman rank correlation coefficient were used to describe the relevance and correlations between species pairs. The results showed that W (48.61), a statistical index to test the variance ratio (VR=0.61), fell outside of the range of Chi-square test, indicating that the overall correlation of all vegetation species was significantly negative. According to the environment adaptation mode of dominant species and the main influencing factors, the species were divided into 4 ecological groups, i.e., Phragmites australis, Carex scabrifolia-Scirpus triqueter - Juncellus serotinus, Spartina alterniflora - Scirpus mariqueter, Echinochloa crusgalli - Imperata cylindrica, based on the ranking of Spearman correlation coefficient. The inter-specific relationships in the salt marsh plant community of southern Chongming Dongtan were complicated and extremely unstable with species sensitive to environmental impacts. According to the analysis of relationships between the species and their pre-sent distribution, we suggested using S. mariqueter as target species to provide strategies for protecting native species based habitats.

Use of Amniotic Microparticles Coated With Fibroblasts Overexpressing SDF-1α to Create an Environment Conducive to Neovascularization for Repair of Full-Thickness Skin Defects.

As angiogenesis and vasculogenesis involve the complex network structures of various types of cells, extracellular matrix components, and cytokines, it is still difficult to exactly mimic the microenvironment of vascularization in vivo. In our study, we constructed a complex containing highly proliferative fibroblasts that can secrete extracellular matrix components and growth factors to chemotaxize endothelial progenitor cells (EPCs) in an attempt to create an ideal microenvironment for quick vascularization. Amniotic membrane microparticles (mAM) rich in type IV collagen (COL IV) and laminin (LN) were prepared, and human dermal fibroblasts (HDF) were infected with lentivirus (LV) of overexpression of SDF-1α to construct SDF-1α(ov)HDF. Using the rotary cell culture system (RCCS), mAM was loaded with HDF or SDF-1α(ov)HDF to construct HDF-mAM and SDF-1α(ov)HDF-mAM complexes. The complexes were able to secrete various types of active peptides (IL-6, IL-8, TGF-ß, and bFGF) during in vitro culture. In addition, SDF-1α(ov)HDF-mAM complex highly expressed SDF-1α. Transwell assay showed SDF-1α(ov)HDF-mAM complex had an apparent chemotactic effect on EPCs. Transplantation of complexes onto full-thickness skin defects of C57BL mice further demonstrated that SDF-1α expression and the number of peripheral EPCs at days 3, 5, and 7 in the SDF-1α(ov)HDF-mAM group were significantly higher than that in other groups (p < 0.01). The local microvascular density at day 10 of transplantation showed that the microvascular density in the SDF-1α(ov)HDF-mAM group was significantly higher than that in HDF-mAM group (p < 0.01). In conclusion, HDF-mAM had a strong proliferative activity and could be used to create a sound microenvironment for quick vascularization by secreting multiple cytokines and extracellular matrix components. Overexpression of SDF-1α could chemotaxize EPCs to reach local wounds, thus further accelerating angiogenesis in the transplant site. The technique described may prove to be a new model for accelerating vascularization of tissue and organ transplants and chronic ischemic wounds.

Developing Shingles-Induced Koebner Phenomenon in a Patient With Psoriasis: A Case Report.

Both shingles and psoriasis are common cutaneous diseases. About 25% of the psoriatic patients develop Koebner phenomenon (KP) after various injuries, and in rare instance, KP may occur at the site of healed or healing shingles.We report a 30-year-old man with 7-month history of scalp psoriasis who developed KP at the areas of developing shingles. Cutaneous examination revealed scaly erythematous papules and plaques located on the scalp and forehead, and groups of clustered erythematous papules with silver scales in the dermatome distributed on the right side of chest wall the prior herpes zoster lesions involved. After removal of the scales on the papules, underlying bleeding points were present.The lesions on chest had good response to anti-psoriatic therapies, as the lesions on scalp did. After a year of follow-up, recurrent psoriasis occurred, but the lesions were located only on the scalp, and the areas of prior occurrence of shingles, because of which we considered diagnosis of recurrent psoriasis rather than relapsing KP for the chest lesions.Not only the healing and healed shingles can trigger KP in psoriasis, but also the developing shingles can cause psoriatic KP at the site of herpes zoster lesions.

Activity of the novel dual phosphatidylinositol 3-kinase/mammalian target of rapamycin inhibitor NVP-BEZ235 against osteosarcoma.

Recent studies have identified that constitutively active phosphatidylinositol 3-kinase (PI3K)/Akt/mammalian target of rapamycin (mTOR) signaling is an important feature of osteosarcoma, where it promotes cell proliferation, survival, and chemo-resistance. Here, we studied the therapeutic potential of NVP-BEZ235, a novel dual PI3K/mTOR dual inhibitor, on osteosarcoma cells in vivo and in vitro. NVP-BEZ235 was cytotoxic and cytostatic to a panel of osteosarcoma lines (MG-63, U2OS and SaOs-2), where it induce apoptosis and cell-cycle arrest. At the molecular level, NVP-BEZ235 inhibited PI3K-AKT-mTORC1 activation and downregulated cyclin D1/cyclin B1 expressions, while increasing MEK/Erk phosphorylation in osteosarcoma cells. MEK/Erk inhibitors PD98059 and MEK-162 increased NVP-BEZ235 activity on osteosarcoma cells. In vivo, oral NVP-BEZ235 inhibited U2OS xenograft in SCID mice, and its anti-tumor efficiency was further enhanced by MEK-162 co-administration. Taken together, our findings indicate that dual inhibition of PI3K and mTOR with NVP-BEZ235, either alone or in combination with MEK/Erk inhibitors, may be an efficient treatment for osteosarcoma.

Bufotalin-induced apoptosis in osteoblastoma cells is associated with endoplasmic reticulum stress activation.

The search for novel and more efficient chemo-agents against malignant osteoblastoma is important. In this study, we examined the potential anti-osteoblastoma function of bufotalin, and studied the underlying mechanisms. Our results showed that bufotalin induced osteoblastoma cell death and apoptosis in dose- and time-dependent manners. Further, bufotalin induced endoplasmic reticulum (ER) stress activation in osteoblastoma cells, the latter was detected by the induction of C/EBP homologous protein (CHOP), phosphorylation of inositol-requiring enzyme 1 (IRE1) and PKR-like endoplasmic reticulum kinase (PERK), as well as caspase-12 activation. Conversely, the ER stress inhibitor salubrinal, the caspase-12 inhibitor z-ATAD-fmk as well as CHOP depletion by shRNA significantly inhibited bufotalin-induced osteoblastoma cell death and apoptosis. Finally, by using a mice xenograft model, we demonstrated that bufotalin inhibited U2OS osteoblastoma cell growth in vivo. In summary, our results suggest that ER stress contributes to bufotalin-induced apoptosis in osteoblastoma cells. Bufotalin might be investigated as a novel anti-osteoblastoma agent.

Assessment of coronary stents by 64-slice computed tomography: in-stent lumen visibility and patency.

OBJECTIVE: To assess lumen visibility of coronary stents by 64-slice computed tomography (CT) coronary angiography, and determine the value of 64-slice CT in non-invasive detecting of in-stent restenosis after coronary artery stent implantation. METHODS: Totally, 60 patients (54 males, aged 57.0+/-12.7 years) and 105 stents were investigated by 64-slice CT at a mean interval of 20.0+/-16.6 months after coronary stents implantation. Axial multi-planar reconstruction images of the stents and curved-planar reconstruction images through the median of the stents were reconstructed for evaluating stent image quality on a 5-point scale (1=excellent, 5=non-assessable), and stent lumen diameter was detected. Conventional coronary angiography was performed in 18 patients, and 32 stents were evaluated. RESULTS: Image quality was good to excellent on average (score 1.71+/-0.76). Stent image quality score was correlated to heart rate (r=0.281, P<0.01) and stent diameter (r=-0.480, P<0.001). All the stents were assessable in lumen visibility with an average visible lumen diameter percentage of 60.7%+/-13.6%. Visible lumen diameter percentage was correlated to heart rate (r=-0.193, P<0.05), stent diameter (r=0.403, P<0.001), and stent image quality score (r=-0.500, P<0.001). Visible lumen diameter percentage also varied depending on the stent type. In comparison with the conventional coronary angiography, 4 of 6 in-stent stenoses were correctly detected. The sensitivity and specificity for the detection of in-stent stenosis were 66.7% and 84.6%, respectively. CONCLUSIONS: Using a 64-slice CT, the stent lumen is partly visible in most of the stents. And 64-slice CT may be useful in the assessment of stent patency.

[Radiological features of dual-energy CT lung perfusion imaging in patients with acute pulmonary embolism: comparison with CT pulmonary angiography].

OBJECTIVE: To explore the relationship of the perfusion defects in the dual-energy CT lung perfusion image and the filling defects in the CT pulmonary angiography (CTPA) in patients with acute pulmonary embolism. METHODS: The clinical data of 22 patients with acute pulmonary embolism were retrospectively analyzed. All of them underwent contrast CT scan in two phases using dual-energy CT: the first is pulmonary artery phase, and the second was performed immediately after the first phase using dual-energy scan covered the whole lung. Perfusion defects in the lung perfusion images were compared with the filling defects in the CTPA. RESULTS: Complete filling defects of segmental and subsegmental pulmonary arteries mostly showed correspondence perfusion defects in the CT perfusion map, accounting for 83% and 62% respectively. However, when there were partial or central filling defects, most of them were partial perfusion defects or normal in the CT perfusion map. Three segmental perfusion defects were depicted without the visualization of endoluminal thrombi within the corresponding arteries. CONCLUSIONS: The perfusion defects in the CT lung perfusion image are not completely corresponding to the filling defects in the CTPA. The combination of CTPA and CT lung perfusion map will offer more information for the acute pulmonary embolism.

Application of computed tomographic colonography in diagnosis of colonic polyps.

OBJECTIVE: To assess the clinical values of computed tomographic colonography (CTC) in diagnosis of colonic polyps. METHODS: Forty-two patients who were clinically suspicious of colonic polyps or underwent colonic polyps screening received examinations with both CTC and conventional colonoscopy. Sixteen- or 64-slice spiral computed tomography and professional imaging processing techniques were used for evaluation. Per-polyp and per-patient results were analyzed. Those by per-polyp were subsequently divided into > or = 10 mm group, 5-10 mm group, and < or =5 mm group. Sensitivity, positive predictive value (PPV), specificity, negative predictive value (NPV), and accuracy were calculated using statistical method for diagnostic studies, with conventional colonoscopy as a gold standard. RESULTS: Ninety and 61 polyps were found by CTC and conventional colonoscopy, respectively. The per-polyp sensitivity/PPV were 80.3%/55.6% in total, and 100%/92.9%, 93.8%/65.2%, and 68.8%/ 41.5% in the > or = 10 mm group, 5-10 mm group, and < or =5 mm group, respectively. The per-patient sensitivity, PPV, specificity, NPV, and accuracy were 97.1%, 89.5%, 42.9%, 75.0%, and 88.1%, respectively. CONCLUSION: CTC can clearly reveal the morphology of colonic polyps and be used as a routine monitoring method for the clinical diagnosis of polyps.

[CT perfusion in evaluation of cerebral hemodynamics in carotid artery stenosis].

OBJECTIVE: To study the clinical value of multi-slice spiral CT (MSCT) perfusion technique in evaluation of cerebral hemodynamics in patients with severe carotid stenooclusive lesions. METHODS: Twenty-three patients with severe carotid stenosis were enrolled for evaluation of cerebral hemodynamics, including 12 patients with carotid occlusion and 11 with severe carotid stenosis (> 70%). Among them, 16 patients had multiple arterial diseases, including contralateral carotid stenosis, vertebral arteries stenosis, and cerebral arteries stenosis or occlusion. All patients received CT perfusion examination. Ten normal persons received CT perfusion were regarded as the control group. RESULTS: Cerebral perfusion was bilaterally symmetrical in 10 normal persons. While in the patients group, cerebral hemodynamic impairments ipsilateral to the stenosis side were found by CT perfusion in 17 patients, including an delay of time to peak (TTP) (17 patients), increase of cerebral blood volume (CBV) (7 patients), and decrease of cerebral blood flow (CBF) (4 patients). No significant difference was found between occlusion group (n = 8) and stenosis group (n = 9) (P = 0.640), but incidences of hemodynamic impairment were significantly higher in multiple arterial diseases group (n = 14) than single arterial disease group (n = 3) (P = 0.045). CONCLUSION: CT perfusion technique can sensitively reveal the cerebral hemodynamic impairments in patients with severe carotid stenooclusive diseases, and therefore valuable for clinical application.

[Application of multi-slice spiral CT in the diagnosis of pulmonary hypertension].

OBJECTIVE: To evaluate the utility of multi-slice spiral CT in the diagnosis of pulmonary hypertension. METHODS: The clinical and multi-slice spiral CT data of 22 patients with pulmonary hypertension were retrospectively analyzed. The diagnoses included chronic thromboembolic pulmonary hypertension (CTEPH) (n = 9), primary pulmonary hypertension (PPH) (n = 7), connective tissue diseases (n = 2), portal-pulmonary hypertension (n = 2), eosinophilia-myalgia syndrome (n = 1), and fibrosing mediastinitis (n = 1). Lung parenchyma, such as ground-glass opacity, mosaic attenuation and septal thickness, mediastinum, pulmonary vascular, right ventricular enlargement and interventricular septum displacement were evaluated on CT. RESULTS: CT pulmonary angiography (CTPA) accurately diagnosed 95% of pulmonary hypertension and the signs of right ventricular dysfunction. Pulmonary embolism was found in 9 patients. The incidence of mosaic attenuation was significantly higher in CTECH than in PPH (P = 0.034). CONCLUSION: CTPA can accurately diagnose pulmonary hypertension and thereby provide useful information for the etiological diagnosis.

Using of multislice helical CT colonography in patients with malignant lesions of colon.

OBJECTIVE: To investigate the techniques and clinical applications of multislice helical computed tomography (CT) colonography in colonic lesions. METHODS: Fifty-nine patients with malignant lesions of colon underwent volume scanning using multislice helical CT. Four types of reconstruction including CT virtual colonoscopy (CTVC), shaded surface display (SSD), Raysum, and multiple planar reconstruction (MPR) were used for image post-processing. The results were compared with those of colonoscopy and pathology. RESULTS: Multislice helical CT colonography detected 54 colorectal carcinomas, 4 adenomas with focal carcinoma, 1 non-Hodgkin's lymphoma (NHL). The lesions' number, size, location, morphology, stricture of intestinal cavity, infiltration, and metastasis were shown satisfactorily by multislice helical CT colonography. Whole colon could be shown in all patients. CT colonography displayed 4 synchronous colonic tumors, 1 ascending colon carcinoma combined with left renal carcinoma among 54 patients with colonic carcinomas. The accuracy of location of CT colonography was 100%. There were 9 cases that CT showed the tumor location was different from the finding of conventional colonoscopy, while all of the CT location were proven exact by operation. CT colonography also displayed the infiltration of serous layer and fatty tissue in 45 cases; 21 cases matched the pathological results in all the 24 cases of suspicious lymph node metastasis, the sensitivity was 87.5%, the specificity was 90.6%; 9 cases hepatic metastasis, 2 ovarian metastasis, and 1 double adrenal gland metastasis. CONCLUSIONS: Multislice helical CT colonography is effective in preoperative diagnosis, location, stage, and making treatment plan of colorectal carcinoma. It can display the portion not seen during colonoscopy and may have an adjunctive role.

[Techniques and clinical applications of 16 multislice helical CT in colonic diseases].

OBJECTIVE: To evaluate the techniques and clinical applications of 16 multislice helical CT in colonic lesions. METHODS: Eighty-one patients including 54 colorectal carcinomas, 5 adenomas, 1 non-Hodgkin's lymphoma, 6 inflammatory bowel diseases, and other 15 cases underwent volume scanning using 16 multislice helical CT. Four types of reconstruction included multiple planar reconstruction, shaded surface display, raysum, and CT virtual colonoscopy. RESULTS: Complete colon could be shown in all patients. The lesions' morphology, number, size, location, intestinal cavity, pericolonic changes, and other abdominal organs were satisfactorily shown by CT. CONCLUSIONS: Sixteen multislice helical CT colonography is a valuable imaging technique for detecting colonic diseases. It is effective in diagnosis and treatment planning. It can display the portions of colon that is inaccessible at colonoscopy.