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Taking squash (Cucurbita pepo L.) variety Alan as test object, this paper studied the effects of exogenous Ca2+ on the morphological and photosynthetic characteristics and chlorophyll fluorescent parameters of squash seedlings under the cross-stress of high temperature and strong light. Under the stress, applying 5-20 mmol∙L-1 of Ca2+ increased the plant height, leaf area, chlorophyll and carotenoid contents, photosynthetic rate (Pn), stoma conductance (Gs), transpiration rate(Tr ), maximal PS-efficiency (Fv / Fm), actual PS-efficiency (ΦPS II), and photochemical queching coefficient (qP ), and decreased the intercellular CO2 concentration (Ci ) and non-photochemical fluorescence quenching coefficient (NPQ), suggesting that this application of exogenous Ca2+ could effectively mitigate the damage of high temperature and strong light stress on the squash seedlings leaf, and make it keep more rapid photosynthetic electron transfer rate and higher PS-electron transfer activity. Among the treatments of applying Ca2+, 10 mmol Ca2+∙L-1 had the best effect. When the Ca2+ application rate exceeded 40 mmol∙L-1, no mitigation effect was observed on the high temperature and strong light stress. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effects of silent information regulator 1 (SIRT1) in amygdala on depression-like behaviors in rats using chronic restraint stress (CRS) as a model of depression., Methods: Sixty male SD rats were randomly divided into six groups ( n =10 per group): control group (Control), chronic restraint stress group (CRS), CRS + fluoxetine-treated group (CRS + FLU), CRS + saline-treated group (CRS + NaCl), CRS + SIRT1-overexpression group (CRS + AAV-SIRT1), and CRS + empty vector group (CRS + AAV-EGFP). Except for the control group, rats from the other groups were exposed to chronic restraint stress for 21 days. After the modeling, rats in fluoxetine-treated group and saline-treated group were, respectively, treated with fluoxetine (10 mg/kg) or saline (10 mg/kg) by gavage every day for 3 weeks; AAV-SIRT1 or AAV-EGFP was, respectively, stereotaxically injected into the amygdala of rats in SIRT1-overexpression group and empty vector group, and the virus was expressed for 3 weeks. Rats in normal control group and CRS model group were not given any drug treatment. The depression-like behaviors of rats in each group were evaluated by sugar preference test (SPT), open field test (OFT) and forced swimming test (FST). SIRT1 expression in amygdala of rats was assessed by using immunoblot blotting. The number of SIRT1-positive cells in amygdala of rats was detected by immunofluorescence technique., Results: Compared with the normal control group, the level of SIRT1 protein and the number of SIRT1 + cells in amygdala of the CRS-exposed rats were decreased significantly ( P ＜0.01), and CRS-exposed rats showed a significant decrease in sucrose preference ( P ＜0.01), less total horizontal distance ( P ＜0.01) and less time entered the center field ( P ＜0.01) in the OFT, a significant increase in the immobility time of the FST ( P ＜0.01). Fluoxetine treatment ( P ＜0.05, P ＜0.01) or SIRT1 overexpression ( P ＜0.01) partially reversed the down-regulation of SIRT1 protein and SIRT1 + cells in amygdala of CRS-exposed rats and significantly improved the depression-like behaviors of CRS rats., Conclusion: Fluoxetine treatment partially reversed the down-regulation of SIRT1 level and the number of SIRT1 + in CRS rats, and significantly improved the depression-like behaviors. The antidepressant effect of fluoxetine treatment may be related to the up-regulation of SIRT1 in the amygdala of CRS-exposed rats.

Since synthetic vitamins were used to fortify food and as supplements in the late 1930s, vitamin intake has significantly increased. This has been accompanied by an increased prevalence of obesity, a condition associated with diabetes, hypertension, cardiovascular disease, asthma and cancer. Paradoxically, obesity is often associated with low levels of fasting serum vitamins, such as folate and vitamin D. Recent studies on folic acid fortification have revealed another paradoxical phenomenon: obesity exhibits low fasting serum but high erythrocyte folate concentrations, with high levels of serum folate oxidation products. High erythrocyte folate status is known to reflect long-term excess folic acid intake, while increased folate oxidation products suggest an increased folate degradation because obesity shows an increased activity of cytochrome P450 2E1, a monooxygenase enzyme that can use folic acid as a substrate. There is also evidence that obesity increases niacin degradation, manifested by increased activity/expression of niacin-degrading enzymes and high levels of niacin metabolites. Moreover, obesity most commonly occurs in those with a low excretory reserve capacity (e.g., due to low birth weight/preterm birth) and/or a low sweat gland activity (black race and physical inactivity). These lines of evidence raise the possibility that low fasting serum vitamin status in obesity may be a compensatory response to chronic excess vitamin intake, rather than vitamin deficiency, and that obesity could be one of the manifestations of chronic vitamin poisoning. In this article, we discuss vitamin paradox in obesity from the perspective of vitamin homeostasis.

Phosphatidylethanolamine N-methyltransferase (PEMT) plays a critical role in breast cancer progression. However, the epigenetic mechanism regulating PEMT transcription remains largely unknown. Here, we show that the first promoter-specific transcript 1 is the major PEMT mRNA species, and methylation of the -132 site is a key regulatory element for the PEMT gene in BRCA1-mutated breast cancer. Mechanistically, hypermethylated -132 site-mediated loss of active histone marks H3K9ac and increase of repressive histone marks H3K9me enrichment synergistically inhibited PEMT transcription. Clinicopathological data indicated that a hypermethylated -132 site was associated with histological grade (P = 0.031) and estrogen receptor status (P = 0.004); univariate survival and multivariate analyses demonstrated that lymph node metastasis was an independent and reliable prognostic factor for BRCA1-mutated breast cancer patients. Our findings imply that genetic (e.g., BRCA1 mutation) and epigenetic mechanisms (e.g., DNA methylation and histone modifications) are jointly involved in the malignant progression of PEMT-related breast cancer.

Objective: To investigate the effect of saponin from Tupistra chinensis Baker (STCB) on lethal toxicity of endotoxin in mice and explore the underlying mechanism., Methods: Mouse models of endotoxin-induced death and endotoximia were established by intraperitoneal administration of KM mice with lipopolysaccharides (LPS) from Pseudomonas aeruginosa in doses of 60 mg/kg and 10 mg/kg respectively. Mouse survival rate and survival time were recorded and the serum levels of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) in endotoximia mice were measured by enzyme-linked immunosorbent assay (ELISA). Mouse peritoneal exudate cells induced by LPS were used as an in vitro inflammatory model,which was then intervened with STCB and the levels of IL-1beta and TNF-alpha in the culture supernatants were measured by ELISA., Results: The survival rates of mice prophylactically treated with STCB (200 and 400 mg/kg, in 5 consecutive days) were slightly higher compared with that in model group,but no statistical difference was observed (P>0.05). The survival time was much longer in the treated group (P<0.05). The serum levels of IL-1beta and TNF-alpha in STCB-treated mice (200 and 400 mg/kg, in 5 consecutive days) were significantly lower compared with those in model group (P<0.05). STCB (20 and 40 microg/mL) remarkably inhibited LPS-induced IL-1beta and TNF-alpha production by peritoneal exudate cells in vitro (P<0.05)., Conclusion: Saponin from Tupistra chinensis showed beneficial effect on the prevention of mice from lipopolysaccharides-induced death, in which down regulation of IL-1beta and TNF-alpha expression might be involved.

Objective: To investigate the effect of prostaglandin E2 (PGE(2)) on the proliferation of cultured hepatocellular carcinoma cells and explore which subtypes of EP prostanoid receptor mediate the action., Methods: RT-PCR was used to determine COX-2 and EP receptor mRNA expression levels in human hepatocellular carcinoma cell line Hep3B and human normal hepatocyte line QSG7701. Cell counting kit-8 (CCK-8) assay was employed to investigate the effect of PGE(2), selective EP2 receptor agonist butaprost and EP3/EP4 receptor agonist PGE1 alcohol on the proliferation of the cells., Results: COX-2 mRNA was highly expressed in Hep3B cells but scarcely in QSG7701 cells. Hep3B cells expressed the mRNAs for all the EP receptor subtypes, but EP2 and EP4 receptors were much more strongly expressed than EP1 and EP3 receptors. PGE(2) significantly promoted Hep3B cell proliferation in a time- and dose-dependent manner, and 10 µmol/L PGE(2) increased the cell proliferation by 22.57% (P<0.001) after a 48-h incubation; treatment with 0.1, 1.0, and 10 µmol/L PGE(2) for 72 h resulted in significantly increased cell proliferation by 12.13% (P<0.01), 17.58% (P<0.01) and 33.07% (P<0.001), respectively. EP2 receptor agonist butaprost (20 µmol/L) increased Hep3B cell proliferation by 21.96% (P<0.001), but the EP3/EP4 receptor agonist PGE(1) alcohol (2-20 µmol/L) exhibited no significant mitogenic effect in Hep3B cells, and 200 µmol/L PGE(1) alcohol decreased the cell viability., Conclusion: Selective activation of EP2 receptor promotes Hep3B cell proliferation, indicating the predominant role of EP2 receptor in mediating the mitogenic effect of PGE2.

Objective: To study the optimal extraction process of immune active polysaccharides from Fomes fomentarius by introduction of ultrasonication., Methods: An orthogonal experimental design of L9 (3(4)) was used to investigate the effects of ultrasonication time, extraction temperature and extraction time on the extraction ratio, sugar content and immune stimulating activity (mouse splenocyte metabolic activity measured with MTT colorimetry) of the polysaccharides and the optimal extraction process was evaluated., Results: Ultrasonication treatment had the most remarkable effect on the immune stimulating activity of the polysaccharides. The optimal extraction process for extraction of immune active polysaccharides was as follows: ultrasonication for 30min, extraction temperature at 80 degrees C and water extraction time for 2h., Conclusion: Ultrasonication can be used as a useful technique for extraction of immune active polysaccharides from Fomes fomentarius.

Taking squash (Cucurbita pepo L.) variety Alan as test object, this paper studied the effects of exogenous Ca2+ on the morphological and photosynthetic characteristics and chlorophyll fluorescent parameters of squash seedlings under the cross-stress of high temperature and strong light. Under the stress, applying 5-20 mmol x L(-1) of Ca2+ increased the plant height, leaf area, chlorophyll and carotenoid contents, photosynthetic rate (Pn), stoma conductance (Gs), transpiration rate (Tr), maximal PS II efficiency (Fv/Fm), actual PS II efficiency (phi(PS II)), and photochemical queching coefficient (q(P)), and decreased the intercellular CO2 concentration (Ci) and non-photochemical fluorescence quenching coefficient (NPQ), suggesting that this application of exogenous Ca2+ could effectively mitigate the damage of high temperature and strong light stress on the squash seedlings leaf, and make it keep more rapid photosynthetic electron transfer rate and higher PS II electron transfer activity. Among the treatments of applying Ca2+, 10 mmol Ca2+ x L(-1) had the best effect. When the Ca2+ application rate exceeded 40 mmol x L(1), no mitigation effect was observed on the high temperature and strong light stress.

Objective: To investigate the effects of matrine on proliferation, cell cycle, apoptosis rate and beta-catenin-dependent transcriptional activity in cultured hepatoma cell line Hep3B., Methods: Cell viability was analyzed by cell counting kit-8 (CCK-8) assay. Cell cycle and apoptosis rate wene determined by flow cytometry analysis, beta-catenin-dependent transcriptional activity was assayed with Dual-Luciferase Reporter System., Results: 72 h of matrine (50 - 800 mg/L) exposure significantly suppresses Hep3B cells growth in a concentration-dependent manner (P<0.01), the 50 percent inhibitory concentration (IC50) was 312.53 mg/L. A higher proportion of apoptotic cells in matrine-treated group than in control groups (21.73 +/- 1.66% vs. 4.39 +/- 1.93%) are shown. Cell proportions in G0/G1 phase were respectively 74.48% and 57.39% in matrine-treated group and control group. Cell proportions in S phase wene respectively 12.94% and 27.67%. Beta-catenin reporter activity was also decreased by matrine treatment in a concentration-dependent manner in Hep3B cells (P<0.05 or P<0.01)., Conclusion: Through inhibition of beta-catenin-dependent transcription, matrine induces cell cycle arrest and apoptosis, and subsequently suppresses proliferation of hepatoma cells.

Objective: To establish a mouse model of humoral immune response by immunization with rabbit red blood cells (RRBCs)., Methods: The mice were immunized with RRBCs and the serum hemolysin level was measured by micro-hemolysis spectrophotometry., Results: The peak time needed for hemolysin production against RRBCs was 6 days after the immunization, and 20% RRBCs in a total volume of 0.2 ml was optimal for intraperitoneal injection. Hydrocortisone (25 mg/kg) and cyclophosphamide (20 mg/kg) inhibited hemolysin production. Mannatide (4 mg/kg) produced no significant effect on serum hemolysin level in normal mice, but significantly potentiated hemolysin production in immunosuppressed mice induced by cyclophosphamide (20 mg/kg)., Conclusion: Intraperitoneal RRBC injection is feasible for establishing mouse models of humoral immune response.

Objective: To observe the effects of lipopolysaccharides of Bacterium prodigiosum (BP-LPS) in inhibiting tumor growth and improving immunosuppression in mice., Methods: In mice bearing S180 tumor and a mouse model of immunosuppression induced by cyclophosphamide (CTX), the tumor growth, indexes of the immune organs and peripheral white blood cell count were measured after intraperitoneal injection of BP-LPS., Results: Injections of BP-LPS (40 U/kg) for 8 consecutive days resulted in a significant inhibition of the tumor growth in mice bearing S180 tumor (P<0.01), with a dose-dependent increase of the spleen indexes but no obvious changes in the thymus indexes. Intraperitoneal injections of BP-LPS for 7 days inhibited the reduction of peripheral white blood cells and spleen indexes in immunosuppressive mice, but did not produce any significant changes in normal mice., Conclusion: BP-LPS can inhibit the tumor growth in tumor-bearing mice and enhance the immune functions of immunosuppressive mice.

Objective: To study the effect of Flammulina velutipes polysaccharides (FVP) on the production of tumor necrosis factor alpha (TNF-alpha), interferon-gamma (INF-gamma) and interleukin 2 (IL-2) by murine immunocytes., Methods: The cell's metabolic activity was determined with methylthiazolyl tetrazolium (MTT) colorimetry assay and the amounts of TNF-alpha, INF-gamma and IL-2 were measured by ELISA., Results: FVP (200, 100, 50 microg/mL) could promote the metabolic activity of murine splenocytes and peritoneal exudate cells (PEC) and increase the amounts of TNF-alpha, INF-gamma and IL-2 in the supernatants of splenocyte cultures, and the amount of TNF-alpha in PEC cultures, with the most marked increase on TNF-alpha level. FVP (100, 50, 25 mg/kg) could raise the serum levels of TNF-alpha and INF-gamma in S180 tumor-bearing mice., Conclusion: FVP may regulate murine immune function through promoting the production of TNF-alpha, INF-gamma and IL-2.

Objective: To investigate the immunomodulatory effects of Fomes fomentarius polysaccharides (FFP) in mice., Methods: MTT assay was employed to evaluate the in vitro metabolic activity of the mouse splenocytes treated with FFP at different concentrations, and the secretion of tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (INF-gamma) and interleukin 2 (IL-2) from the cells were measured by enzyme-linked immunosorbent assay. The changes in the phagocytotic activity of mouse macrophage in response to FFP treatment were evaluated by phagocytosis percentage of chicken red blood cells (CRBCs). The effect of FFP on the humoral immunity was assessed in mice immunized with sheep red blood cells (SRBCs) by measuring the serum levels of specific antibody (hemolysin) against SRBCs., Results: FFP at the concentrations of 25, 50, and 100 microg/ml all significantly enhanced the metabolic activity of mouse splenocytes in vitro and increased the production of TNF-alpha, IFN-gamma and IL-2. FFP treatment also markedly enhanced the metabolic activity of mouse peritoneal exudate cells and TNF-alpha production by the cells. At the doses of 25, 50, and 100 mg/kg, FFP significantly increased serum hemolysin level in mice immunized with SRBCs, and FFP at 50 and 100 mg/kg obviously increased the capacity of mouse peritoneal macrophages in vivo for CRBC phagocytosis., Conclusion: FFP can promote the secretion of TNF-alpha, IFN-gamma and IL-2 by mouse immunocytes and enhance mouse humoral immune response and the phagocytotic activity of the macrophages.

Objective: To study the anticoagulant activity of different portions of leech ethanol extracts (LEEs)., Methods: Anticoagulant activity was determined by measuring prothrombin time(PT), thrombin time (TT), activated partial throboplstin time (APTT) and fibrinogen coagulation time (FCT)., Results: PT, TT, APTT and FCT were remarkably prolonged by ethyl acetate portion of LEEs. Portions of petroleum ethrer, n-butanol and water extracted from LEEs was much weaker on anticoagulant activity., Conclusion: The anticoagulant effect of ethyl acetate extract portion of LEEs is the strongest among the four portions, and this results may be from its inhibitory effect on thrombin-catalyzed fibrinogen hydrolysis.

Objective: To investigate the effects of cyclooxygenase inhibitor diclofenac on the proliferation and cyclooxygenase-2 (COX-2) mRNA expression of cultured hepatocellular carcinoma cell lines HepG2, Hep3B and human hepatocellular cell line QSG-7701., Methods: After exposure to diclofenac at various concentrations (10-200 micromol/L) for 24, 48 and 72 h, the cell proliferation was analyzed by Cell Counting Kit-8 (CCK-8) assay and mRNA expression determined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR)., Results: Diclofenac exposure for 24, 48 and 72 h significantly inhibited HepG2 and Hep3B cell proliferation in a concentration-dependent manner, with inhibition rate of 40.47% and 54.49% after 48 h exposure to 50 micromol/L diclofenac and IC50 of 70.54 and 48.39 micromol/L, respectively. A much weaker antiproliferative effect on QSG-7701 cells was shown, with IC50 of 189.91 micromol/L after 48-hour exposure to diclofenac. RT-PCR detected COX-2 mRNA in HepG2 and Hep3B cells, but hardly in QSG-7701 cells. Treatment with diclofenac or 5-Fu resulted in elevated COX-2 mRNA expression both in HepG2 and Hep3B cells., Conclusion: Diclofenac can specifically inhibit the proliferation of COX-2-expressing HepG2 and Hep3B cells, and induce up-regulation of COX-2 mRNA expression, which indicates the important role of COX-2 in the proliferation of hepatoma cells.

Objective: To determine the content of dexamethasone sodium phosphate in Fuyankang cream, a preparation for treatment of various skin conditions., Methods: Dexamethasone sodium phosphate content in Fuyankang cream was determined using high-performance capillary electrophoresis (HPCE) at the detection wavelength of 254 nm, under the optimal condition achieved with a fused-silica capillary tube (60 cm x 75 microm) and 100 mmol/L sodium tetraborate buffer (pH 9.2) at a constant voltage of 25 kV with a sampling time of 10 s at 25 degrees Celsius;., Results: The calibration curve displayed good linear relationship within dexamethasone sodium phosphate concentration range of 20 to 100 microg/ ml r=0.999 8 . The average recovery of dexamethasone sodium phosphate was 99.75% n=5, RSD=1.03% ., Conclusion: HPCE is simple, quick and sensitive in determination of dexamethasone sodium phosphate content in Fuyankang cream.