Opportunistic use of lumbar computed tomography and magnetic resonance imaging for osteoporosis screening.

Compared with the healthy patients, patients with osteoporosis had a lower Hounsfield unit (HU) value and a higher vertebral bone quality (VBQ) score. Both the HU value and VBQ score can simply distinguish patients with osteoporosis (OP), with a cutoff value of HU value < 97.06 and VBQ score > 3.08. INTRODUCTION: The purpose of this study is to determine whether the opportunistic use of computed tomography (CT) or magnetic resonance imaging (MRI) is effective for identifying spine surgical patients with OP. METHODS: We retrospectively evaluated 109 lumbar spine surgery patients who received lumbar quantitative CT (QCT) and MRI. Using the area under the curve, the CT-based HU value and MRI-based VBQ score were calculated. Then, based on the QCT results, receiver operating characteristic (ROC) curves were constructed to determine the diagnostic performance of the HU value and VBQ score. RESULTS: The HU value was significantly lower in the OP group, and the VBQ score was significantly higher in the OP group. Using the area under the curve, the diagnostic performance of the HU value and VBQ score for OP were 0.959 and 0.880, respectively. The diagnostic threshold values determined with optimal sensitivity and specificity were an HU value of 97.06 and a VBQ score of 3.08. CONCLUSION: Opportunistic use of CT and MRI can simply distinguish patients with OP, which are expected to be potential alternatives to T-score for the osteoporosis screening.

DEPDC1 affects autophagy-dependent glycolysis levels in human osteosarcoma cells by modulating RAS/ERK signaling through TTK.

The current treatment for osteosarcoma (OS) is based on surgery combined with systemic chemotherapy, however, gene therapy has been hypothesized to improve patient survival rates. The density-enhanced protein domain 1 protein (DEPDC1) functions as a crucial determinant in the advancement of OS, which is highly expressed in OS cells. The current study was designed to delve into the effect and mechanism of DEPDC1 and phosphotyrosine-picked threonine tyrosine kinase (TTK) in OS. The expression of DEPDC1 and TTK in OS cells was detected by western blotting. Furthermore, the assessment of glycolysis encompassed the quantification of extracellular acidification rate, glucose uptake rate, lactate concentration, and the expression of glucose transporter 1, hexokinase 2, and pyruvate kinase M2. Finally, the functions of DEPDC1 and TTK in autophagy and ras-extracellular signal-regulated kinase signaling were determined by western blotting after interfering with DEPDC1 in SaOS-2 cells. The results revealed that DEPDC1 and TTK were upregulated in OS cell lines and interfering with DEPDC1 inhibited glycolysis and autophagy in OS cells. Furthermore, the STRING database suggested that DEPDC1 and TTK perform targeted binding. Notably, the results of the present study revealed that DEPDC1 upregulated RAS expression through TTK and enhanced ERK activity, thereby affecting glycolysis and autophagy in OS cells. Collectively, the present investigation demonstrated that DEPDC1 affected autophagy-dependent glycolysis levels of OS cells by regulating RAS/ERK signaling through TTK.

Anandins A and B, Two Rare Steroidal Alkaloids from a Marine Streptomyces anandii H41-59.

Anandins A (1) and B (2), two rare steroidal alkaloids, were isolated from the fermentative broth of a marine actinobacteria Streptomyces anandii H41-59. The gross structures of the two alkaloids were elucidated by spectroscopic methods including HR-ESI-MS, and NMR. Their absolute configurations were confirmed by single-crystal X-ray diffraction analysis and comparison of their experimental and calculated electronic circular dichroism spectra, respectively. Anandin A exhibited a moderate inhibitory effect against three human cancer cell lines MCF-7, SF-268, and NCI-H460 with IC50 values of 7.5, 7.9, 7.8 µg/mL, respectively.

[Non-alkaloid Chemical Constituents from Macleaya cordata].

Objective: To study the non-alkaloid chemical constituents of Macleaya cordata. Methods: Alcohol extraction and liquidliquid partitionmethods were used to extract the chemical constituents. Silica gel,reverse-phase octadecylsilyl( ODS), and Sephadex LH-20 column chromatographic methods were applied for isolation and purification. Spectroscopic methods including MS and NMR were used to determine their structures. Results: Eleven non-alkaloid compounds were isolated from the fruits of Macleaya cordata, and their structures were identified as 3-( 3,4-dihydroxy) phenylpropanoic acid methyl ester( 1),ferulic acid( 2),1-octacosanol( 3),syringic acid( 4),p-hydroxy-benzoic acid( 5),p-coumaric acid( 6),quercetin-3-O-ß-D-glucoside( 7),N-p-coumaroyl tyramine( 8),10-eicosenoic acid( 9) and ß-sitosterol( 10) and daucosterol( 11). Conclusion: Compounds 1,3 ~9 are isolated from Macleaya cordata for the first time.

A new flavonoid glucoside from Rhododendron seniavinii.

The leaves of Rhododendron seniavinii Maxim with little phytochemical information are used as folk remedies for the treatment of acute and chronic bronchitis in China. In our pursuing for the biologically active chemical constituents in the leaves, a new flavonoid glycoside 5,7,3'-trimethoxy-quercetin-3-O-ß-D-glucopyranoside (1) was isolated from the water extract of its leaves, together with two known compounds 5,7,3'-trimethoxy-quercetin (2) and ovafolinin B-9'-O-ß-D-glucopyranoside (3). The structures of the new flavonoid glucoside as well as two known compounds were elucidated by spectroscopic and chemical methods.

A new cytotoxic benzophenanthridine isoquinoline alkaloid from the fruits of Macleaya cordata.

A new cytotoxic benzophenanthridine isoquinoline alkaloid, named cordatine (1), together with one known alkaloid 8-methoxydihydrochelerythrine (2), was isolated from the fruits of Macleaya cordata. The structure of the new compound was elucidated by spectroscopic methods including 1D and 2D NMR, HR-ESI-MS. Both compounds indicated significant cytotoxicity against MCF-7 and SF-268 cell lines.

[Alkaloids from Macleaya cordata and their cytotoxicity assay].

OBJECTIVE: To study the alkaloids of Macleaya cordata and their anti-tumor activities. METHOD: Alcohol and liquid-liquid extraction were used methods were used to extract the alkaloids constituents, and silica gel, reverse-phase octadecylsilyl (ODS), sephadex LH-20 chromatographic methods and HPLC were applied to isolate and purify compounds. MS, NMR spectroscopic methods were used to determine their structures. Furthermore, the cytotoxicity of these chemical components for MCF-7 and SF-268 cell lines was measured by MTT method. RESULT: Twelve alkaloids were isolated from the fruits of M. cordata, and their structures were identified as: maclekarpine E (1), 6-acetonyldihyrochelerythrine (2), cavidilinine (3), 6-acetonyldihyrosanguinnarine (4), O-methylzanthoxyline (5), 6-methoxy-dihydrosanguinarine (6), spallidamine (7), 6-hydroxyldihydrochelerythrine (8), arnotianamida (9), dihydrosanguinarine (10), protopine (11), and cryptopine (12). CONCLUSION: Compounds 1, 3, 7-9 were isolated from M. cordata for the first time, and compound 5 is a new natural product. The results of cytotoxic assay indicated that compound 6 showed strong cytotoxicity against MCF-7 and SF-268 cell lines with IC50 values of 0.61 µmol · L(-1) and 0.54 µmol · L(-1), respectively.

A polymethoxyflavone from Laggera pterodonta induces apoptosis in imatinib-resistant K562R cells via activation of the intrinsic apoptosis pathway.

BACKGROUND: Treatment with imatinib mesylate (IM) (a tyrosine kinase inhibitor) is the first line of standard care for patients newly diagnosed with CML. Despite the success of IM and other tyrosine kinase inhibitors (TKIs), chronic myeloid leukemia (CML) remains largely incurable, and a number of CML patients die due to Abl mutation-related drug resistance and blast crisis. 3, 5-Dihydroxy-6, 7, 3'4'-tetramethoxyflavone (DHTMF) is a polymethoxyflavone isolated from Laggera pterodonta which is a herbal medicine used to treat cancer in the Chinese folk. In the previous study, we found DHTMF demonstrated good antiproliferative activities against a number of cancer cell lines and induced the apoptosis of CNE cells in vitro in a time- and dose-dependent manner while exhibiting low cytotoxicity in the two normal cell lines Vero and EVC304. The aim of the present study was to evaluate the proliferation inhibition and apoptosis induced by DHTMF alone and in combination with IM in the IM-resistant CML cell line K562R. METHODS: Cell proliferation was assayed with the cell counting kit-8 (CCK8) method. The apoptosis percentage was determined by flow cytometry (FCM). Mitochondrial transmembrane potential was detected using FCM and confocal laser-scanning microscopy. The level of proteins involved in apoptosis was detected by Western blotting. RESULTS: DHTMF suppressed K562R cell viability in both time- and dose-dependent manners. DHTMF combined with IM enhanced the inhibitory effects and apoptosis in K562R cells as compared with DHTMF alone. DHTMF alone and in combination with IM significantly decreased the mitochondrial membrane potential and increased the levels of cleaved caspase-9, caspase-7, caspase-3, and PARP in K562R cells. CONCLUSIONS: We demonstrated that DHTMF could inhibit IM-resistant K562R cell proliferation and induces apoptosis via the intrinsic mitochondrial apoptotic pathway. These results suggest that DHTMF may be a potential therapeutic drug with lower side effects against IM resistance in CML cells.

Two new acetylated flavonoid glycosides from Phyllanthus urinaria.

Two new acetylated flavonoid glycosides, quercetin 3-O-α-l-(2,4-di-O-acetyl) rhamnopyranoside-7-O-α-l-rhamnopyranoside (1) and quercetin 3-O-α-l-(3,4-di-O-acetyl) rhamnopyranoside-7-O-α-l-rhamnopyranoside (2), together with two known compounds, quercetin (3) and quercetin 3-O-α-l-rhamnopyranoside (4), were isolated from the ethanol extract of Phyllanthus urinaria. The structures of the new compounds were determined on the basis of extensive spectroscopic data including IR, HR-ESI-MS, 1D NMR, and 2D NMR.

[Studies on the chemical constituents from Conyza canadensis].

OBJECTIVE: To study the chemical constituents of Conyza canadensis. METHODS: Chromatographic methods including HP20 macroporous resin, silica gel, Sephadex LH-20 and eighteen alkyl silane bonding silica gel (ODS) were used for the isolation and purification of Conyza canadensis. The structures of the obtained compounds were identified by physical chemistry and spectroscopic data. RESULTS: Six compounds were isolated from ethanol-extraction of water area of C. canadensis and identified as Eugenyl beta-Psd (1), scutellarin (2), luteolin-7-O-beta-D-glucuronide (3), quercetin (4), quercetin-3-O-beta-D-glucopyranoside (5) and luteolin (6). CONCLUSION: Compounds 1,3,5 and 6 are isolated from C. canadensis for the first time.

Comparison of the differentiation abilities of bone marrow-derived mesenchymal stem cells and adipose-derived mesenchymal stem cells toward nucleus pulposus-like cells in three-dimensional culture.

Nucleus pulposus cell (NPC) transplantation can be a potential therapeutic approach for intervertebral disc degeneration (IDD). However, low cell viability has restricted the therapeutic capacity of NPCs, and sources of natural NPCs are limited. Bone marrow-derived mesenchymal stem cells (BMSCs) and adipose-derived mesenchymal stem cells (ADSCs) can be differentiated toward NPC-like cells. However, it is unknown whether there are differences in the abilities of these two cell types to differentiate into NPC-like cells, or which cell type exhibits the best differentiation ability. The present study compared the abilities of BMSCs and ADSCs to differentiate toward NPC-like cells with or without a 3D culture system to lay a foundation for stem cell transplantation therapy for IDD. BMSCs were isolated from the rat whole bone marrow cell using the repeated adherent culture method. ADSCs were isolated from rat adipose tissues in the subcutaneous inguinal region using the enzyme digestion method. Cells were identified using flow cytometry. Cell viability was assessed via Cell Counting Kit-8 assays, and reverse transcription-quantitative PCR and western blotting were carried out to evaluate the expression of NPC markers and chondrocyte-specific genes. Glycosaminoglycans (GAGs) and proteoglycans were examined via Alcian blue and safranin O staining, respectively. ADSCs in 3D culture displayed the highest cell proliferative ability, compared with the 2D culture system and BMSC culture. In addition, ADSCs in 3D culture exhibited increased GAG and proteoglycan synthesis than BMSCs. Compared with BMSCs in 3D culture, ADSCs in 3D culture exhibited higher mRNA and protein expression of NPC marker genes (hypoxia-inducible factor 1-α, glucose transporter 1) and chondrocyte-specific genes (Sox-9, aggrecan and type II collagen). The present findings indicated that ADSCs exhibited a better ability to differentiate into NPC-like cells in 3D culture compared with BMSCs, which may be of value for the regeneration of intervertebral discs using cell transplantation therapy.

[Research on antiproliferative effect of flavones isolated from Laggera pterodonta].

OBJECTIVE: To research the cytotoxicity and in vitro antiproliferative effect of the six flavone compounds extracted from Laggera pterodonta. METHOD: The cytotoxicity on the normal cells and antiproliferative effect on tumor cells were tested by MTT assay, and then the preliminary structure-activity relationship was analysed. The phase distribution of the cell cycle and apoptosis rate were analyzed by flow cytometry. RESULT: The results of MTT assay showed 5,7,3',4'-tetramethoxy-3-hydroxyflavone and chrysosplenetin B inhibited growth of A549 and Hela cells significantly with a dose dependent mode, while exhibited low cytotoxicity to the two normal cells, Vero and EVC304. Both compounds contain ortho-phenolic methoxyl moietys in their structures. Flow cytometry analysis revealed that Hela cells treated with increasing quantities of chrysosplenetin B increased the percentage of cells in the G2/M phase, and Hela and A549 cells treated with increasing quantites of the 5,7,3',4'-tetramethoxy-3-hydroxyflavone and chrysosplenetin B increased the apoptosis rates. CONCLUSION: The 5,7,3',4'-tetramethoxy-3-hydroxyflavone and chrysosplenetin B extracted from L. pterodonta showed high antiproliferative effect on cancer cells with low cytotoxicity on normal cells, and took the effects on A549 and Hela cells through the hold-up of the G2/M phase of the cell cycle and induction of the apoptosis.

[Chemical constituents of Laggera pterodonta].

OBJECTIVE: To study the chemical constituents of Laggera pterodonta. METHOD: The ethanol extract of L. pterodonta was isolated by column chromatogramphy on silica gel, ODS, and Sephadex LH-20 to afford compounds. The structures of the obtained compounds were identified by chemical reactions and spectroscopic analysis. RESULT: Nineteen compounds were separated and identified to be pterodondiol (1), ilicic acid (2), artemitin (3), chrysosplenetin B (4), 3,5-dihydroxy-3',4',6,7-tetramethoxyflavone (5), chrysosplenol D (6), 5,6,4'-trihydroxy-3,7-dimethoxyflavone (7), quercetin (8), tamarixetin (9), patuletin (10), quercetin-3-O-beta-D-galactopyranoside (11), patuletin-3-O-beta-D-glucopyranoside (12), helichrysoside (13), 4,5,7-trihydroxy-6-methoxyflavone-3-O-beta-D-rutinoside (14), kaempferol-3-O-beta-D-glucopyranoside (15), stigmasterol (16), stigmasterol 3-O-beta-D-glucopyranoside (17), 2-hydroxy-benzoic acid (18), beta-sitosterol (19). CONCLUSION: Compounds 5, 7, 9-15, and 17-18 were isolated from this plant for the first time. The 13C-NMR data of compound 7 is reported for the first time.

Sasanquasaponin ΙΙΙ from Schima crenata Korth induces autophagy through Akt/mTOR/p70S6K pathway and promotes apoptosis in human melanoma A375 cells.

BACKGROUND: Melanoma is a high fatality skin cancer which lacks effective drugs. Sasanquasaponin, an important sort of constituents in theaceae, has been demonstrated to have potent anti-tumor effect in breast cancer and hepatocellular carcinoma. As a sasanquasaponin, we speculate that Sasanquasaponin III (SQS III) isolated from Schima crenata Korth may also have anti-tumor activity. PURPOSE: This study aims to investigate whether SQS III has anti-melanoma activity and examine the underlying mechanisms of SQS III against melanoma. METHODS/STUDY DESIGNS: The anti-proliferative effect of SQS III was assessed by cells viability assay. Annexin V-FITC/PI double staining assay was utilized for detection of apoptosis. Mitochondrial membrane potential and reactive oxygen species (ROS) production were detected using JC-1 and DCFH-DA assay, respectively. Autophagy was monitored using transmission electron microscopy (TEM) and GFP-LC3 transfection fluorescence analysis. Autophagosome-lysosome fusion and lysosomal degradation were determined using a GFP-LC3 & LAMP1 co-localization assay and DQ-BSA staining. Proteins related to apoptosis and autophagy were analyzed by Western blotting. RESULTS: Our results demonstrated that the SQS III exhibited potent anti-cancer activity in A375 cells by inducing both apoptosis and autophagy. In melanoma cells treated with SQS III, caspases were activated and PARP was cleaved, proving the occurrence of apoptosis. Mechanistic studies indicated that the pro-apoptosis activity of SQS III was mediated by death receptor pathway and mitochondrial dysfunction which was induced by ROS accumulation and reversed by the ROS inhibitor N-acetyl-cysteine (NAC). In addition to triggering apoptosis, SQS III may also cause autophagy in melanoma cells. Our results demonstrated that SQS III induced up-regulated expression of GFP-LC3, autophagosome-lysosomal fusion and lysosomal degradation. Additionally, the ROS accumulation was also involved in the activation of autophagy. Meanwhile, it was also found that after SQS III treatment, the expression of LC3-II was up-regulated and the AKT/mTOR signaling pathway was inhibited. The autophagy inhibitor 3-MA converted cytotoxicity and apoptosis of SQS III in A375 cells, which indicated that autophagy promoted the SQS III-induced apoptosis. CONCLUSION: SQS III showed potent anti-cancer activity by inducing apoptosis and autophagy, which provides insights into its possible use as a therapy for melanoma.

Two new quercetin glycoside derivatives from the fruits of Gardenia jasminoides var. radicans.

Two new quercetin glycoside derivatives named quercetin-3-O-[2-O-trans-caffeoyl-α-L-rhamnopyranosyl-(1 → 6)-ß-D-glucopyranoside] (1) and quercetin-3-O-[2-O-trans-caffeoyl-ß-L-rhamnopyranosyl-(1 → 6)-ß-D-glucopyranoside] (2) along with three known flavonoids, 5-hydroxy-6,7,3',4',5'-pentamethoxyflavone (3), 5,7-dihydroxy-8-methoxyflavone (4) and kaempferol 3-O-ß-D-glucopyranoside (5), were isolated from the fruits of Gardenia jasminoides var. radicans. The structures of the new compounds were determined by means of extensive spectroscopic analysis (1D, 2D NMR and HR-ESI-MS), glycoside hydrolysis and sugar HPLC analysis after derivatisation. This is the first report on the isolation of a pair of compounds with α or ß-L-rhamnopyranosyl configuration from plant and the first detail assignment of their NMR data.

A new triterpenoid from the fruits of Gardenia jasminoides var. radicans Makino.

A novel triterpenoid 3α,16ß,23,24-tetrahydroxy-28-nor-ursane-12,17,19,21-tetraen (1) was isolated from the fruits of Gardenia jasminoides var. radicans Makino. The structure of the new compounds was elucidated on the basis of spectroscopic analysis including MS and NMR data. Compound 1 was in vitro tested for cytostatic activity on human throat cancer (Hep-2) cell line by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide method and showed mild anticancer activity with the IC50 of 31.2 µM.

Triterpenoid saponins from the root bark of Schima superba and their cytotoxic activity on B16 melanoma cell line.

Eight new oleanane-type triterpenoid saponins, schisusaponins A-H, along with eight known triterpenoid saponins, were isolated from the root bark of Schima superb (Theaceae). Their structures were elucidated on the basis of extensive spectroscopic analyses and chemical methods. The cytotoxicity of the new compounds against B16 melanoma cells was assessed. Among the isolated new saponins, schisusaponins C and E showed more potent effects (with IC50 values of 10.08 and 10.89 µM) than vinblastine (with an IC50 value of 19.48 µM).

Comparison of our self-designed rotary self-locking intramedullary nail and interlocking intramedullary nail in the treatment of long bone fractures.

OBJECTIVE: The purpose of this study is to compare the clinical effects of our self-designed rotary self-locking intramedullary nail (RSIN) and interlocking intramedullary nail (IIN) for long bone fractures. METHODS: A retrospective study was performed in 1,704 patients who suffered bone fractures and underwent RSIN or IIN operation in our hospital between March 1999 and March 2013, including 494 with femoral fractures, 572 with humeral fractures, and 638 with tibial fractures. Among them, 634 patients were followed up for more than 1 year. The operative time, intraoperative blood loss, postoperative complications, healing rate, and the excellent and good rate of functional recovery were compared between two groups. RESULTS: Compared with IIN group, RSIN group exhibited significantly shorter operative time and less intraoperative blood loss no matter for humeral, femoral, or tibial fractures (all p < 0.001). The healing rate in patients with more than 1 year follow-up was significantly higher in RSIN group for femoral and tibial fractures (both p < 0.05). In RSIN group, no nail breakage or loosening occurred, but radial nerve injury and incision infection were respectively observed in one patient with humeral fracture. In IIN group, nail breakage or loosening occurred in 7 patients with femoral fractures and 16 patients with tibial fractures, radial nerve injury was observed in 8 patients with humeral fractures, and incision infection was present in 2 patients with humeral fractures and 1 patient with femoral fracture. The complication rate of IIN group was significantly higher than that of RSIN group (p < 0.05). However, there were no significant differences in the excellent and good rate of shoulder, elbow, knee, and ankle joint functional recovery between RSIN group and IIN group. CONCLUSION: RSIN may be a reliable and practical alternative method for the treatment of long bone fractures.