Combined detection tumor markers for diagnosis and prognosis of gallbladder cancer.

AIM: To clarify the value of combined use of markers for the diagnosis of gallbladder cancer and prediction of its prognosis. METHODS: Serum cancer antigens (CA)199, CA242, carcinoembryonic antigen (CEA), and CA125 levels were measured in 78 patients with gallbladder cancer (GBC), 78 patients with benign gallbladder diseases, and 78 healthy controls using electrochemiluminescence. CA199, CA242, CEA, and CA125 levels and positive rates were analyzed and evaluated pre- and post-operatively. Receiver operator characteristic curves were used to determine diagnostic sensitivity and specificity of GBC. Survival time analysis, including survival curves, and multivariate survival analysis of a Cox proportional hazards model was performed to evaluate independent prognostic factors. RESULTS: Serum CA242, CA125, and CA199 levels in the GBC group were significantly higher when compared with those in the benign gallbladder disease and healthy control groups (P < 0.01). With a single tumor marker for GBC diagnosis, the sensitivity of CA199 was the highest (71.7%), with the highest specificity being in CA242 (98.7%). Diagnostic accuracy was highest with a combination of CA199, CA242, and CA125 (69.2%). CA242 could be regarded as a tumor marker of GBC infiltration in the early stage. The sensitivity of CA199 and CA242 increased with progression of GBC and advanced lymph node metastasis (P < 0.05). The 78 GBC patients were followed up for 6-12 mo (mean: 8 mo), during which time serum CA199, CA125, and CA242 levels in the recurrence group were significantly higher than in patients without recurrence (P < 0.01). The post-operative serum CA199, CA125, and CA242 levels in the non-recurrence group were significantly lower than those in the GBC group (P < 0.01). Multivariate survival analysis using a Cox proportional hazards model showed that cancer of the gallbladder neck and CA199 expression level were independent prognostic factors. CONCLUSION: CA242 is a marker of GBC infiltration in the early stage. CA199 and cancer of the gallbladder neck are therapeutic and prognostic markers.

[Effect of Roux-en-Y gastric bypass on quality of life in non-obese patients with type 2 diabetes mellitus].

OBJECTIVE: To observe the impact of Roux-en-Y gastric bypass on quality of life in non-obese patients with type 2 diabetes mellitus. METHODS: Thirty-seven non-obese patients with type 2 diabetes mellitus who underwent Roux-en-Y gastric bypass were prospectively studied. A 36-item short form healthy survey questionnaire(SF36), the diabetes treatment satisfaction questionnaire(DTSQ), and quality of life scale for patients with type 2 diabetes mellitus(DMQLS) were used to evaluate the quality of life for all the non-obese patients with type 2 diabetes mellitus preoperatively and 12 months postoperatively. RESULT: The blood glucose and lipid indexes were significantly decreased after operation(all P<0.05). SF36 showed the physical and mental synthesis scores at 12 month after operation were 74.6±18.3 and 79.8±14.9 respectively, higher than those at one week before operation(54.9±15.1 and 56.4±17.8, both P<0.01). DTSQ showed treatment satisfaction score was increaced significantly after operation(29.2±7.1 vs. 15.4±5.6, P<0.01). The quality of life evaluated by DMQLS, was also significantly improved(P<0.01). CONCLUSION: Roux-en-Y gastric bypass can significantly improve the quality of life for non-obese patients with type 2 diabetes mellitus.

Targeting of the Sonic Hedgehog pathway by atractylenolides promotes chondrogenic differentiation of mesenchymal stem cells.

Molecules that enhance chondrogenic differentiation in mesenchymal stem cells (MSCs) were identified and isolated using an in vitro Gli reporter gene assay in MSCs incorporating a Sonic Hedgehog (Shh) target. Atractylenolide III, which promoted Gli1-mediated transcriptional activity, was isolated from an ethyl acetate extract of the Rhizoma, Atractylodis macrocephalae. After dehydration, atractylenolide III was transformed to atractylenolide I. Both atractylenolides were confirmed by MS, UV, IR, 1H- and 13C-NMR spectra. Atractylenolide III (which contains -OH at the 8-position) and atractylenolide I (which lacks -OH at the 8-position) were found to effectively promote the activity of the Gli promoter. While the hydroxyl group of atractylenolide III was not essential for the effect of atractylenolide, its effect was dependent on Shh signaling. Phenotypic cellular analysis indicated that atractylenolides induced MSCs to differentiate into chondrocytes, as shown by increased expression of specific chondrogenic markers including collagen II, aggrecan and the cartilage related transcription factor, Sox9. Atractylenolides significantly increased the expression of Shh and its target gene Gli-1, indicating that Shh signaling was activated by atractylenolides. Moreover, inhibition of Shh signaling reduced the effect of atractylenolides on the chondrogenic phenotype. The discovery that atractylenolides induce chondrocytes from MSCs is promising for bony disease therapy.

Chemical components in extracts from Plastrum testudinis with proliferation-promoting effects on rat mesenchymal stem cells.

Plastrum Testudinis (PT) is often used as an important traditional Chinese medicine to treat bone diseases in China for centuries. To identify active components of PT involved in promoting proliferation of MSCs, PT was extracted with ethyl acetate and separated by silica gel column chromatography with gradient elution. Sixteen (Ts-1 ≈ 16) components were obtained, which were biologically evaluated by MTT assay and flow cytometry on the proliferation of rat marrow-derived mesenchymal stem cells (rMSCs). Results indicated that Ts-12 could induce the proliferation compared with control group (p < 0.05), while Ts-4 exhibited inhibitive effect. The chemical components of PT which regulated the proliferation of rMSCs were investigated by Gas Chromatography-Mass Spectrometry (GC-MS), HPLC, and nine standard compounds. The experimental results suggested that palmitic acid methyl ester and cholesterol myristate, which identified from Ts-12, possessed proliferative activity while stearic acid found in Ts-4 showed inhibition.

Caveolin-1 plays a crucial role in inhibiting neuronal differentiation of neural stem/progenitor cells via VEGF signaling-dependent pathway.

In the present study, we aim to elucidate the roles of caveolin-1(Cav-1), a 22 kDa protein in plasma membrane invaginations, in modulating neuronal differentiation of neural progenitor cells (NPCs). In the hippocampal dentate gyrus, we found that Cav-1 knockout mice revealed remarkably higher levels of vascular endothelial growth factor (VEGF) and the more abundant formation of newborn neurons than wild type mice. We then studied the potential mechanisms of Cav-1 in modulating VEGF signaling and neuronal differentiation in isolated cultured NPCs under normoxic and hypoxic conditions. Hypoxic embryonic rat NPCs were exposed to 1% O2 for 24 h and then switched to 21% O2 for 1, 3, 7 and 14 days whereas normoxic NPCs were continuously cultured with 21% O2. Compared with normoxic NPCs, hypoxic NPCs had down-regulated expression of Cav-1 and up-regulated VEGF expression and p44/42MAPK phosphorylation, and enhanced neuronal differentiation. We further studied the roles of Cav-1 in inhibiting neuronal differentiation by using Cav-1 scaffolding domain peptide and Cav-1-specific small interfering RNA. In both normoxic and hypoxic NPCs, Cav-1 peptide markedly down-regulated the expressions of VEGF and flk1, decreased the phosphorylations of p44/42MAPK, Akt and Stat3, and inhibited neuronal differentiation, whereas the knockdown of Cav-1 promoted the expression of VEGF, phosphorylations of p44/42MAPK, Akt and Stat3, and stimulated neuronal differentiation. Moreover, the enhanced phosphorylations of p44/42MAPK, Akt and Stat3, and neuronal differentiation were abolished by co-treatment of VEGF inhibitor V1. These results provide strong evidence to prove that Cav-1 can inhibit neuronal differentiation via down-regulations of VEGF, p44/42MAPK, Akt and Stat3 signaling pathways, and that VEGF signaling is a crucial target of Cav-1. The hypoxia-induced down-regulation of Cav-1 contributes to enhanced neuronal differentiation in NPCs.

(E)-Ethyl 2-cyano-3-[4-(4,5-diphenyl-1H-imidazol-2-yl)phen-yl]acrylate dihydrate.

In the title compound, C(27)H(21)N(3)O(2)·2H(2)O, the three benzene rings attached to the heterocyclic imidazole ring are not coplanar with the latter, making dihedral angles of 14.8 (2), 31.4 (2), and 37.5 (2)°, respectively, for the benzene ring planes in the 2-, 4- and 5-positions. In the crystal, there are two water mol-ecules which serve as connectors between the acrylate mol-ecules and stabilize the structure via N-H⋯O, O-H⋯N, C-H⋯O and O-H⋯O hydrogen bonding.

Identification of polymer materials using laser-induced breakdown spectroscopy combined with artificial neural networks.

A combination of laser-induced breakdown spectroscopy (LIBS) and artificial neural networks (ANNs) has been used for the identification of polymer materials, including polypropylene (PP), polyvinyl chloride (PVC), polytetrafluoroethylene (PTFE), polyoxymethylene (POM), polyethylene (PE), polyamide or nylon (PA), polycarbonate (PC) and poly(methyl methacrylate) (PMMA). After optimization of the experimental setup and the spectrum acquisition protocol, successful identification rates between 81 and 100% were achieved using spectral features gathered from single spectra without averaging (1 second acquisition time) over a wide spectral range (240-820 nm). Furthermore, ten different materials based on PVC were tested using the identification procedure. Correct identifications were obtained as well. Sorting of the materials into sub-categories of PVC materials according to their charges (concentration in trace elements such as Ca) was performed. The demonstrated capacities fit, in practice, the needs of plastic-waste sorting and of producing high-grade recycled plastic materials.

BMP-Id pathway targeted by cholesterol myristate suppresses the apoptosis of PC12 cells.

Identifying small molecules that suppress apoptosis is promising for the therapy of brain diseases. We recently showed that autocrine bone morphogenetic protein (BMP) signaling involves the effects of cholesterol myristate present in traditional Chinese medicine on mesenchymal stem cells. The present study evaluated the effects of cholesterol myristate on the apoptosis and BMP signaling of PC12 cells. PC12 cells transfected by the inhibitor of differentiation (Id1) promoter reporter construct target gene of BMP4 signaling; cholesterol myristate increases the activity of Id1 promoter. However, structurally related steroids such as cholesterol, ß-sitosterol and cholesten-3-one, lack of the myristate, did not affect the activity of Id1 promoter, suggesting that myristate is essential for the effect of cholesterol myristate. These effects depend on BMP signaling. Apoptosis analysis indicated that cholesterol myristate inhibited the apoptosis of PC12 cells induced in serum-free condition. Cholesterol myristate significantly increases the expression of BMP4, BMPRIA, p-Smad1/5/8, Id1 and its antiapoptotic target gene Bcl-xL in PC12 cells treated in serum-free condition. Moreover, BMP antagonist reduced the anti-apoptotic effect of cholesterol myristate. Thus, this study is to provide evidence that BMP-Id pathway targeted by cholesterol myristate suppresses the apoptosis of PC12 cells. Our findings are therefore of considerable therapeutic significance and provide the potential of newly exploiting cholesterol myristate and clinically in brain disease therapies.

Cholesterol myristate suppresses the apoptosis of mesenchymal stem cells via upregulation of inhibitor of differentiation.

To identify small molecules that suppress the apoptosis of mesenchymal stem cells (MSCs) is promising for stem cell therapy. We recently showed that bone morphogenetic protein 4 (BMP4) signalling involves the effect of cholesterol myristate on the proliferation of MSCs. The present study evaluated the effects of cholesterol myristate on the apoptosis of MSCs and the inhibitor of differentiation (Id1), target gene of BMP4 signalling. MSCs transfected by the Id1 promoter reporter construct, cholesterol myristate increases the activity of Id1 promoter. However, structurally related steroids such as cholesterol, ß-sitosterol and cholesten-3-one, lack of the myristate, did not affect the activity of Id1 promoter, suggesting that myristate is essential for this effect. This effect depends on BMP signalling. Apoptosis analysis indicated that cholesterol myristate inhibited the apoptosis of MSCs induced by serum-free. Cholesterol myristate increases the expression of Id1 and its target gene bcl-x/l in MSCs treated with serum-free. Moreover, noggin, a BMP antagonist, reduced the anti-apoptotic effects of cholesterol myristate. Thus, this study aims to provide evidence that cholesterol myristate suppresses the apoptosis of MSCs via up-regulation of Id1. These findings can be applied for improving MSCs survival in stem-cell transplantation, bone-marrow transplantation, treatment of bone diseases such as osteoporosis and chemotherapy.

(+)-Cholesten-3-one induces differentiation of neural stem cells into dopaminergic neurons through BMP signaling.

To identify small molecules that induce dopaminergic neurons from neural stem cells (NSCs) is promising for therapy of Parkinson's disease. Here we report the results of analyzing structurally related steroids in traditional Chinese medicine to identify agents that enhance dopaminergic differentiation of NSCs. Using P19 cells transfected by tyrosine hydroxylase (TH) promoter reporter construct, (+)-Cholesten-3-one with carbonyl, but not cholesterol and cholesterol myristate can effectively promote the activity of TH promoter. This effect depends on bone morphogenetic protein (BMP) signaling. Phenotypic cellular analysis indicated that (+)-Cholesten-3-one induces differentiation of NSCs to dopaminergic neurons with increased expression of specific dopaminergic markers including TH, dopamine transporter, dopa decarboxylase and higher level of dopamine secretion. (+)-Cholesten-3-one significantly increases the expression of BMPR IB, but not BMPR IA or BMPR II; p-Smad1/5/8 positive nuclei and expression of p-Smad1/5/8 were detected in NSCs treated with (+)-Cholesten-3-one, indicating that (+)-Cholesten-3-one may activate the BMP signaling. Moreover, overexpression of BMP4 or inhibition of BMP affects the effect of (+)-Cholesten-3-one on the dopaminergic phenotype. These findings may contribute to efficient production of dopaminergic neurons from NSCs culture for many applications and raise interesting questions about the role of (+)-Cholesten-3-one in neurogenesis.

Hexadecanoic acid from Buzhong Yiqi decoction induced proliferation of bone marrow mesenchymal stem cells.

Buzhong Yiqi decoction (BYD) is a well-known ancient tonic prescription in traditional Chinese medicine (TCM). The purpose of this study is to identify active components of BYD involved in promoting proliferation of mesenchymal stem cells (MSCs) and to investigate its mechanism. BYD was extracted with petroleum ether, ethanol, and water. Evidence provided by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, bromodeoxyuridine, proliferation cell nuclear antigen immunoreactivity, cell cycle analysis, and gas chromatography-mass spectrometry indicated that hexadecanoic acid (HA) in BYD extracted with petroleum ether is the active compound responsible for increasing proliferation of MSCs. Western blot analysis show that HA significantly increase retinoic acid receptor (RAR) levels of MSCs, but not estrogen receptor, thyroid hormone receptor, vitamin D receptor, glucocorticoid receptor, and peroxisome proliferator-activated receptor. Reverse transcription-polymerase chain reaction revealed that HA significantly increased RAR mRNA levels. Furthermore, the mechanism of HA action depends on RAR pathway and up-regulates expression of mRNA for insulin-like growth factor-I, the target gene of RAR. Our findings have now allowed for a refinement in our understanding of TCM with respect to pharmacological regulation of stem cells and may be useful to stem cell biology and therapy.

In vitro studies on the antioxidant and protective effect of 2-substituted -8-hydroxyquinoline derivatives against H(2)O(2)-induced oxidative stress in BMSCs.

Novel 2-vinyl-8-hydroxyquinoline derivatives as potential antioxidants and regulators of H(2)O(2)-induced oxidative stress in rat bone marrow mesenchymal stem cells (MSCs) are first reported. The antiradical properties and the reducing power of these compounds were assessed using 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and auto-oxidation of pyrogallol method, respectively. The activity against lipid peroxidation was determined using ammonium thiocyanate method. The results revealed that introduction of electron-donating groups at 2nd position decreased the antioxidant activities of 8-hydroxyquinoline derivatives. In addition, compound 4, the structure of which is similar to melatonin, exhibited superior antioxidant activities in scavenging DPPH free radical, O(2) free radical, and anti-LPO activities. Except for compounds 7, 12, and 15, the other compounds exhibited a stimulatory effect on MSCs growth. Using hydrogen peroxide (H(2)O(2)), we also investigated the protective efficacy of 2-vinyl-8-hydroxyquinoline derivatives against oxidative stress-induced cell death of MSCs. Cell viability assayed by MTT method indicated that exposure of MSCs cultures to hydrogen peroxide resulted in a concentration-dependent decrease in cell viability, and compounds 4 and 5 at given concentration (2.62 x 10(-3) m) could protect MSCs against H(2)O(2)-induced oxidative stress in bone mesenchymal stem cell (BMSCs).

Autocrine BMP4 signaling involves effect of cholesterol myristate on proliferation of mesenchymal stem cells.

We recently identified that cholesterol myristate in traditional Chinese medicine (TCM) is the active compound that increases proliferation of mesenchymal stem cell (MSCs). The present study is further to determine what signal pathway involves in effect of cholesterol myristate. Reverse transcription-PCR, Western blot and ELISA analysis show that cholesterol myristate increases the release of bone morphogenetic protein 4 (BMP4) from MSCs and the expression in the intracellular levels of BMP4 in a time- and dose dependent manner. However, structurally related steroids such as cholesterol and cholesten presented in TCM, both lack of the myristate, did not affect the secretion and expression of BMP4 on MSCs. These finds suggest that myristate is essential for the effects of cholesterol myristate. Furthermore, cholesterol myristate significantly increase BMPRIB levels of MSCs and the number of BMPRIB positive cells in a time- and dose dependent manner, but not BMPR IA or BMPR II. Our results indicate that action of cholesterol myristate may activate the BMP4-BMPRIB autocrine. Moreover, a blocking antibody against BMP4 or the BMP4 antagonist, noggin, partially reduced the effects of cholesterol myristate on MSCs proliferation. Thus, this study is to provide evidence that autocrine BMP4 signaling involves effect of cholesterol myristate on MSCs proliferation.

Synthesis and evaluation of novel substituted 5-hydroxycoumarin and pyranocoumarin derivatives exhibiting significant antiproliferative activity against breast cancer cell lines.

A library of novel 5-hydroxycoumarin and pyranocoumarin derivatives was constructed via silica sulfuric acid-catalyzed pechmann reaction and Pd(0)-catalyzed suzuki coupling in tandem, and their antiproliferative activities against breast cancer cells MCF-7 and MDA-MB-231 were evaluated. The results showed that compounds such as 6b, 6d, 6h, and 6k possess significant antiproliferative activity against MCF-7 cell line with the IC(50) values of 7.2, 5.3, 3.3, and 6.5 microM, respectively.

Characterization of chemical components in extracts from Si-wu decoction with proliferation-promoting effects on rat mesenchymal stem cells.

Si-wu decoction (SDE), a classic prescription in Traditional Chinese Medicine, has been used for the treatment of a variety of anaemia in China for centuries. In order to explore the scientific basis of the formula, we investigated the relationship between its chemical components and proliferation-promoting effects on rat marrow-derived mesenchymal stem cells (MSCs). Twenty (F-1-F-20) components were obtained and their proliferation-promoting effects on MSCs were investigated. The results showed that F-4, F-7, F-10, and F-11 stimulated the proliferation of the MSCs. The chemical components with proliferation-promoting effects on the MSCs were further identified by GC-MS, HPLC, LC-MS, and other spectra. Ligustilide (F-4) isolated from SDE showed the best proliferation-promoting effect. Palmitic acid methyl ester and stearic acid ethyl identified from F-7 and F-10 by HPLC were also confirmed to be responsible for stimulating MSC proliferation. A novel compound, 6,7-dihydroxy-3-(-prop-1-enyl)isobenzofuran-1(3H)-one, was found in the SDE for the first time by LC-MS(n), whose structure was similar to ligustilide.

Experimental long-distance decoy-state quantum key distribution based on polarization encoding.

We demonstrate the decoy-state quantum key distribution (QKD) with one-way quantum communication in polarization space over 102 km. Further, we simplify the experimental setup and use only one detector to implement the one-way decoy-state QKD over 75 km, with the advantage to overcome the security loopholes due to the efficiency mismatch of detectors. Our experimental implementation can really offer the unconditionally secure final keys. We use 3 different intensities of 0, 0.2, and 0.6 for the light sources in our experiment. In order to eliminate the influences of polarization mode dispersion in the long-distance single-mode optical fiber, an automatic polarization compensation system is utilized to implement the active compensation.

[Effect of fatty acids from plastrum testudinis on proliferation of rat bone mesenchymal stem cell].

To investigate the components in Plastrum Testudinis which have effects on the proliferation of rat bone marrow mesenchymal stem cells( bMMSCs), the active parts of plastrum testudinis which can promote proliferation of rat mesenchymal stem cells were extracted by petroleum aether. The activities of inducing the proliferation of bMMSCs were studied by MTT assay and flow cytometry. The chemical components of extraction were analyzed by GC-MS. The results showed that the petroleum aether extraction can obviously promote the proliferation of the stem cells. The main components are long-chain fatty acids, cholesterols and cholest-4-en-3-one, and palmitic acid, stearic acid and cholest-4-en-3-one have effects on proliferation of bMMSCs. In plastrum testudinis, fatty acids can promote the proliferation of bMMSCs but not increase overly. This provide the experiment basis, and offer important reference for Traditional Chinese Medicine that researches stem cells.

[Effect of extract components from Plastrum testudinis and their combination component on proliferation of rat mesenchymal stem cell in vitro].

OBJECTIVE: To observe the effect of extract components from Plastrum Testudinis and their combination component on the proliferaiton of mesenchymal stem cell (MSC) to screen out the effective components. METHODS: Mesenchymal stem cells were dissociated from bone marrow of rat and were marked by Brdu, and the expression of CD44, CD54 and double label of Brdu and CD44 extract components (A to approximately J) with different concentration (2 microl, 5 microl,10 microl,20 microl)added to cultured MSC. The cell viability was tested by MTT. RESULTS: Cell viability in component A, B, H, I with 20 microl, C, F with 10 microl and J with 5 microl concentration group increased compared with that in control group (P < 0.05) and cell viability in compponent E with 2 microl to approximately 20 microl concentration groups decreased compared with that in control group (P < 0.05). Cell viability in component D and G with 2 microl to approximately 20 microl concentration groups was similar to that in control group. Cell viability in combination component with J component increased, but cell viability in combination component with E component decrased. CONCLUSION: All compoents A, B,C,F, H, I and J can improve proliferation of MSC. Anong them, component J and its combination are the best. Component E and its combination can inhibit proliferation. But component D and G have no effect on proliferation of MSC.

8-Hydroxyquinoline derivatives induce the proliferation of rat mesenchymal stem cells (rMSCs).

A series of 8-hydroxyquinoline derivatives with different substituted groups at 2- or 5-position have been synthesized and characterized. Their effects on the proliferation of the rat marrow-derived mesenchymal stem cells (rMSCs) have been evaluated by MTT assay and flow cytometry. We also analyzed the ability of these compounds to regulate the proliferation of rMSCs and the relationship with the structures of 8-hydroxyquinoline. Compounds 8-11, in which, the vinyl-substituents are on the 2-position of 8-hydroxyquinoline, appear to be able to induce the proliferation of rMSCs. These results show that compounds 8-11 provide a kind of new substances for regulating the proliferation of rMSCs.