A monolithic copolymer prepared from N-(4-vinyl)-benzyl iminodiacetic acid, divinylbenzene and N,N'-methylene bisacrylamide for preconcentration of cadmium(II) and cobalt(II) from biological samples prior to their determination by ICP-MS.

A capillary monolith consisting of poly[N-(4-vinyl)-benzyl iminodiacetic acid-co-divinylbenzene-co-N,N'-methylene bisacrylamide), referred to as poly(VBIDA-DVB-Bis), has been prepared. It is shown to be an efficient sorbent for the enrichment of Co(II) and Cd(II). The two ions are completely retained by the monolith in the pH range from 4.0 to 9.0. The breakthrough curve tests were adopted to evaluate the adsorption performance of the monolith towards Co(II) and Cd(II). A dose-response model was used to describe the breakthrough curves of the two ions at different initial concentrations. The adsorption capacities for Co(II) and Cd(II) are 1.54 and 1.73 mg·m-1 at a concentration level of 2.5 mg·L-1, respectively. The enrichment factor is 100, and the required sample volume is 5 mL. Following elution of the two ions with 0.5 M HNO3, they were quantified by ICP-MS. The limits of detection in a 1 mL sample are 0.35 ng·L-1 for Co(II) and 0.44 ng·L-1 for Cd(II). The method was applied to the determination of Co(II) and Cd(II) in spiked rice, human urine and seawater samples. Graphical abstract Schematic representation of a monolithic copolymer prepared from N-(4-vinyl)-benzyl iminodiacetic acid (VBIDA), divinylbenzene (DVB) and N,N'-methylene bisacrylamide (Bis) and its application for selective capturing of cadmium(II) and cobalt(II) from complex sample matrices prior to their determination by ICP-MS.

C1qTNF-related protein-6 protects against doxorubicin-induced cardiac injury.

The clinical use of doxorubicin (DOX) is limited by its toxic effect. However, there is no specific drug that can prevent DOX-related cardiac injury. C1qTNF-related protein-6 (CTRP6) is a newly identified adiponectin paralog with many protective functions on metabolism and cardiovascular diseases. However, little is known about the effect of CTRP6 on DOX-induced cardiac injury. The present study aimed to investigate whether CTRP6 could protect against DOX-related cardiotoxicity. To induce acute cardiotoxicity, the mice were intraperitoneally injected with a single dose of DOX (15 mg/kg). Cardiomyocyte-specific CTRP6 overexpression was achieved using an adenoassociated virus system at 4 weeks before DOX injection. The data in our study demonstrated that CTRP6 messenger RNA and protein expression were decreased in DOX-treated hearts. CTRP6 attenuated cardiac atrophy induced by DOX injection and inhibited cardiac apoptosis and improved cardiac function in vivo. CTRP6 also promoted the activation of protein kinase B (AKT/PKB) signaling pathway in DOX-treated mice. CTRP6 prevented cardiomyocytes from DOX-induced apoptosis and activated the AKT pathway in vitro. CTRP6 lost its protection against DOX-induced cardiac injury in mice with AKT inhibition. In conclusion, CTRP6 protected the heart from DOX-cardiotoxicity and improves cardiac function via activation of the AKT signaling pathway.

Profiling and quantification of aminophospholipids based on chemical derivatization coupled with HPLC-MS.

In this study, a novel strategy based on acetone stable-isotope derivatization coupled with HPLC-MS for profiling and accurate quantification of aminophospholipids (phosphatidylethanolamine and phosphatidylserine) in biological samples was developed. Acetone derivatization leads to alkylation of the primary amino groups of aminophospholipids with an isopropyl moiety; the use of deuterium-labeled acetone (d6-acetone) introduced a 6 Da mass shift that was ideally suited for profiling and quantification analysis with high selectivity and accuracy. After derivatization, significantly increased column efficiency for chromatographic separation and detection sensitivity for MS analysis of aminophospholipids was observed. Furthermore, an accuracy quantification method was developed. Aminophospholipids in biological samples were derivatized with d0-acetone; while more than two aminophospholipid standards were selected for each class of aminophospholipid and derivatized with d6-acetone, which were then used as the internal standards to typically construct a calibration curve for each class to normalize the nonuniformity response caused by the differential fragmentation kinetics resulting from the distinct chemical constitution of individual aminophospholipid species in the biological samples. The excellent applicability of the developed method was validated by profiling and quantification of aminophospholipids presented in liver samples from rats fed with different diets.

Preventive Cold Acclimation Augments the Reparative Function of Endothelial Progenitor Cells in Mice.

BACKGROUND/AIMS: Chronic cold exposure may increase energy expenditure and contribute to counteracting obesity, an important risk factor for cerebrocardiovascular diseases. This study sought to evaluate whether preventive cold acclimation before ischemia onset might be a promising option for preventing cerebral ischemic injury. METHODS: After a 14-day cold acclimation period, young and aged mice were subjected to permanent cerebral ischemia, and histological analyses and behavioral tests were performed. Mouse endothelial progenitor cells (EPCs) were isolated, their function and number were determined, and the effects of EPC transplantation on cerebral ischemic injury were investigated. RESULTS: Preventive cold acclimation before ischemia onset increased EPC function, promoted ischemic brain angiogenesis, protected against cerebral ischemic injury, and improved long-term stroke outcomes in young mice. In addition, transplanted EPCs from cold-exposed mice had a greater ability to reduce cerebral ischemic injury and promote local angiogenesis compared to those from control mice, and EPCs from donor animals could integrate into the recipient ischemic murine brain. Furthermore, transplanted EPCs might exert paracrine effects on cerebral ischemic injury, which could be improved by preventive cold acclimation. Moreover, preventive cold acclimation could also enhance EPC function, promote local angiogenesis, and protect against cerebral ischemic injury in aged mice. CONCLUSIONS: Preventive cold acclimation before ischemia onset improved long-term stroke outcomes in mice at least in part via promoting the reparative function of EPC. Our findings imply that a variable indoor environment with frequent cold exposure might benefit individuals at high risk for stroke.

[Effects of Xiaodu Yuji Paste on Protein Expressions of VEGF/SDF-1 a/CXCR4 in Granulation Tissue of Diabetic Foot Patients].

Objective To observe the effects of Xiaodu Yuji Paste (XYP) on protein expressions of vascular endothelial growth factor (VEGF)/stromal cell derived factor la (SDF-1a)/chemokine recep- tor 4 (CXCR4) in granulation tissue of diabetic foot patients. Methods Totally 56 patients with diabetic foot were assigned to the control group (29 cases) and the treatment group (27 cases) according to Wagner grading method (the range and the degree of foot lesion). Patients in the control group received basic treatment (anti-inflammation, blood glucose control, anti-coagulation, debridement, drainage, and so on) for 8 weeks. Patients in the treatment group additionally received XYP for 8 weeks. The wound healing was observed. Contents and protein expressions of VEGF/SDF-1 a/CXCR4 were detected using SP method and Western blot. Results The wound healing rates after 2, 4, 8 weeks of treatment were signifi- cantly higher in the treatment group than in the control group (all P <0. 05). Contents and protein expres- sions of VEGF/SDF-1 a/CXCR4 protein expression at week 8 after treatment were all significantly higher in the treatment group than in the control group (P <0. 05). Conclusion The therapeutic effect of XYP might be associated with promoting expressions of VEGF/SDF-la/CXCR4, thus promoting angiogenesis and facilitating wound healing.

[Effects of Direct Moxibustion of "Ganshu"(BL 18) on the Contents of T cells in Peripheral Blood in Rats with Precancerous Lesion of Primary Hepatocellular Carcinoma].

OBJECTIVE: To observe the effect of moxibustion stimulation of "Ganshu"(BL 18) region on contents of T cells in the peripheral blood in rats with Diethylnitrosamine (DEN)-induced hepatocellular carcinoma (HCC), so as to explore its effective in improving immunoregulatory function. METHODS: Seventy male Wistar rats were randomly divided into control group (n=10), model group(n=15), direct moxibustion-15 s group(n=15), direct moxibustion-30 s group (n=15) and ginger-separated moxibustion group(n=15). The primary HCC precancerous lesion model was established by intraperitoneal injection of DEN (50 mg/kg), once every 3 days for 10 weeks. Moxibustion was applied to bilateral "Ganshu"(BL 18) for about 15 min (3 moxa-cones), or 30 min (six moxa-cones), with or without ginger-slice separation, once every other day for 10 consecutive weeks. The contents of T cells of CD 3+, CD 4+, CD 8+ and CD 4+/CD 8+ ratio in the peripheral blood were detected with Flow Cytometey(FCM), pathological changes of liver were observed by light microscope after hematoxylin-eosin(HE) stain. RESULTS: Compared to the control group, the contents of blood CD 3+ and CD 4+ T cells and ratio of CD 4+/CD 8+ were significantly down-regulated, while that of CD 8+T cells was obviously increased in the model group(P<0.05,P<0.01). After moxibustion intervention, the decreased CD 3+ and CD 4+T cells and CD 4+/CD 8+ levels and the increased CD 8+T cell contents were reversed in all the 3 moxibustion groups (P<0.05, P<0.01), except CD 3+ in the ginger-separated moxibustion group (P>0.05). There were no significant differences among the three moxibustion groups in the CD 3+, CD 4+, CD 8+ and CD 4+/CD 8+ levels (P>0.05). In addition, the pathological changes of liver tissue as central vein deviation or absence, disordered arrangement of hepatic cords, narrowing of the hepatic sinusoid, hyperplasia of collagen fibers, formation of tuberosis, unevenness of liver cells with nuclear anachromasis and higher heteromorphism, and macronucleus oncocytes in primary HCC rats were not observed or milder after moxibustion intervention. CONCLUSIONS: Direct moxibustion and ginger-separated moxibustion can improve pathological changes of hepatic cells in rats with HCC, which may be associated with its actions in raising blood CD 3+ and CD 4+ T cell contents and reducing CD 8+ T levels to improve immune function.

[Effects of acupuncture stimulation of different acupoint groups on sleeping latency, serum and hippocampal TNF-α and IL-25 contents in rats with gastric mucosal injury].

OBJECTIVE: To observe the effect of acupuncture intervention on gastric ulcer (GU) and sleeping quality from the viewpoint of brain-gut axis which plays an important role in the regulation of many vital functions in the body. METHODS: Forty male Wistar rats were randomized into normal control, GU model, acupuncture of "Zhongwan"(CV 12)-"Zusanli"(ST 36, gastric function regulating acupoints), acupuncture of "Shenmai" (BL 62)-"Zhaohai" (KI 6, sleep-promotion acupoints), and acupuncture of CV 12-ST 36+ BL 62-KI 6 (combined treatment) groups, with 8 rats in each group. GU model was established by intragastric perfusion of dehydrated alcohol (1 mL/rat), and sleep model established by intraperitoneal injection of pentobarbital sodium (40 mg/kg) after the last treatment. The abovementioned acupoints were punctured with filiform needles and stimulated by manipulating the needle for about 30 s, once every 5 min during 20 min of needle retention. The treatment was conducted once daily for five days. The contents of tumor necrosis factor-alpha (TNF-α) and interleukin-25(IL-25) in the serum and hippocampal tissues were detected by ELISA. RESULTS: Compared with the normal control group, the gastric ulcer index score, barbiturate-induced sleeping time, and TNF-α and IL-25 contents in both serum and hippocampus were significantly increased in the model group (P < 0.01). Following acupuncture treatment, in comparison with the model group, the gastric ulcer index score, barbiturate-induced sleeping time, and TNF-α and IL-25 contents in both serum and hippocampus were significantly down-regulated in the CV 12-ST 36, BL 62-KI 6 and combined treatment groups (P < 0.01, P < 0.05). The effects of the CV 12-ST 36 and combined treatment groups were remarkably superior to those of the BL 62-KI 6 group in down-regulating ulcer index score, serum IL-25, and hippocampal TNF-α and IL-25 contents (P < 0.01, P < 0.05). In addition, the effects of the BL 62-KI 6 and combined treatment groups was considerably better than that of the CV 12-ST 36 group in shortening barbiturate-induced sleeping time (P < 0.01, P < 0.05). The effect of the combined treatment group was markedly better than that of the CV 12-ST 36 and BL 62-KI 6 groups in lowering serum TNF-α content (P < 0.05). CONCLUSION: Acupuncture stimulation of CV 12, ST 36, KI 6 and BL 62 can relieve the gastric mucosal lesion, and shorten barbiturate-induced sleeping time in gastric ulcer rats, which may be related to its effects in reducing TNF-α and IL-25 contents in the serum and hippocampus tissues, suggesting a correlation between the gastrointestinal disorder and sleeping.

Involvement of beta-site APP cleaving enzyme 1 (BACE1) in amyloid precursor protein-mediated enhancement of memory and activity-dependent synaptic plasticity.

The amyloid precursor protein (APP) undergoes sequential cleavages to generate various polypeptides, including the amyloid-beta protein (Abeta), which forms amyloid plaques in Alzheimer's disease (AD), secreted APPalpha (sAPPalpha) which enhances memory, and the APP intracellular domain (AICD), which has been implicated in the regulation of gene transcription and calcium signaling. The beta-site APP cleaving enzyme 1 (BACE1) cleaves APP in an activity-dependent manner to form Abeta, AICD, and secreted APPbeta. Because this neural activity was shown to diminish synaptic transmission in vitro [Kamenetz F, Tomita T, Hsieh H, Seabrook G, Borchelt D, Iwatsubo T, Sisodia S, Malinow R (2003) Neuron 37:925-937], the prevailing notion has been that this pathway diminishes synaptic function. Here we investigated the role of this pathway in vivo. We studied transgenic mice overproducing APP that do not develop AD pathology or memory deficits but instead exhibit enhanced spatial memory. We showed enhanced synaptic plasticity in the hippocampus that depends on prior synaptic activity. We found that the enhanced memory and synaptic plasticity are abolished by the ablation of one or both copies of the BACE1 gene, leading to a significant decrease in AICD but not of any other APP cleavage products. In contrast to the previously described negative effect of BACE1-mediated cleavage of APP on synaptic function in vitro, our in vivo work indicates that BACE1-mediated cleavage of APP can facilitate learning, memory, and synaptic plasticity.