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Several near-stoichiometric lithium niobate (LiNbO3) plates were prepared by the vapour transport equilibrium technique. Domain reversal was carried out on these samples by electric field poling. A nonlinear dependence of switching field on the concentration of anti-site niobium () ions was observed. This nonlinear relationship was explained by a two-dimensional model based on the dynamics of the domain wall. A parameter of &thetas;c, denoting the critical position of the domain wall round ions, was introduced to describe domain reversal. The domain wall energy per unit area for near-stoichiometric lithium niobate was also roughly calculated to be larger than 0.27 J m−2. [ABSTRACT FROM AUTHOR]

Ag/cross-linked poly(vinyl alcohol) (PVA) cable-like nanostructureswere synthesized with control in an aqueous solution of a hydrolysableamphiphilic block polymer, poly(vinyl acetone) (PVKA) (ketalization degreeDH = 0.533)under γ-ray irradiation, via one-step in situ reduction ofAg+ and cross-linking of alcohol units. In the present approach, we try to control the speed ofthe cross-linking reaction of PVA chains (alcohol units), which are yielded from thehydrolysed PVKA, utilizing the low hydrolysis rate of the PVKA in dilute acidic solution. [ABSTRACT FROM AUTHOR]

Silver flowerlike and dendritic nanostructures were synthesized in an aqueous solution of ahydrolysable amphiphilic block copolymer polyvinylacetone (PVKA) via in situ reduction ofAg+ and hydrolysis of PVKA at room temperature. Compared with a previous result, thecomplete hydrolysis time of PVKA is greatly shortened in this process. [ABSTRACT FROM AUTHOR]

Introduction: Health literacy refers to acquiring and utilizing health information to make health-related actions and decisions. Filipino domestic workers with low health literacy are often vulnerable to health problems due to poor living and working conditions., Objective: This study examined the effect of an online health literacy promotion program on health literacy and health knowledge of Filipino domestic workers in Macao., Methods: The mixed method combined quantitative findings of a quasi-experimental study with qualitative results applied. The quasi-experiment included three parallel groups: a synchronous online education group (videoconference group), an asynchronous online education group (video group), and a control group. A total of 88 Filipino domestic workers were assigned to one of these groups, and eight respondents participated in two focus-group interviews respectively after the intervention., Results: For quantitative data, both synchronous and asynchronous online education interventions had positive effects. While comparing with the control group, participants in the videoconference group were more likely to have better health promotion health literacy after the intervention (β = 5.36, p  = .02), and participants in the video group were more likely to have better general health literacy (β = 5.17, p  = .01), disease prevention health literacy (β = 5.31, p  = .04), health promotion health literacy (β = 5.97, p  = .01). For qualitative data, three themes and eight subthemes were extracted after the online health literacy promotion program. After integrating the findings of this study, the study found that this program was essential and beneficial for Filipino domestic workers' health knowledge and health literacy., Conclusion: Overall, online health literacy promotion programe had positive impacts on participants revealed in this study. Asynchronous online education has made significant progress in overall health literacy, which may be more suitable as a widely promoted education method because of the characteristics and working conditions of this population., Competing Interests: The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2024.)

In this paper, bis (indol-3-yl) methanes (BIMs) were synthesised and evaluated for their inhibitory activity against α-glucosidase and α-amylase. All synthesised compounds showed potential α-glucosidase and α-amylase inhibitory activities. Compounds 5 g (IC 50 : 7.54 ± 1.10 μM), 5e (IC 50 : 9.00 ± 0.97 μM), and 5 h (IC 50 : 9.57 ± 0.62 μM) presented strongest inhibitory activities against α-glucosidase, that were ∼ 30 times stronger than acarbose. Compounds 5 g (IC 50 : 32.18 ± 1.66 µM), 5 h (IC 50 : 31.47 ± 1.42 µM), and 5 s (IC 50 : 30.91 ± 0.86 µM) showed strongest inhibitory activities towards α-amylase, ∼ 2.5 times stronger than acarbose. The mechanisms and docking simulation of the compounds were also studied. Compounds 5 g and 5 h exhibited bifunctional inhibitory activity against these two enzymes. Furthermore, compounds showed no toxicity against 3T3-L1 cells and HepG2 cells.HighlightsA series of bis (indol-3-yl) methanes (BIMs) were synthesised and evaluated inhibitory activities against α -glucosidase and α-amylase.Compound 5g exhibited promising activity (IC 50 = 7.54 ± 1.10 μM) against α -glucosidase.Compound 5s exhibited promising activity (IC 50 = 30.91 ± 0.86 μM) against α-amylase.In silico studies were performed to confirm the binding interactions of synthetic compounds with the enzyme active site.

An efficient alkylcyanation of in situ generated arynes by N,N-disubstituted aminomalononitriles is described, enabling the direct synthesis of 2-substituted benzonitriles.

A mild and efficient method for the synthesis of cyanoformamides from N,N-disubstituted aminomalononitriles with CsF as the promoter has been developed. This method features a wide substrate scope and high reaction efficiency, and will facilitate corresponding cyanoformamide-based biological studies and synthetic methodology development.

A facile and efficient method to form a chiral multi-substituted 3H-spiro[benzofuran-2,1'-cyclopentane] structural unit has been developed via a one-pot [3+2] cyclization/semipinacol rearrangement cascade. A catalysis system of Cu(ii)/BOX has been used to efficiently construct a key stereogenic center via a cyclization between substituted benzoquinones and allylic alcohols affording the desired products in good yields and with excellent enantioselectivities and diastereoselectivities (21 examples; up to 67% yields; up to 92% ee and up to >20 : 1 dr). This method provides an alternative strategy for the synthesis of the corresponding bioactive molecules containing spiro[benzofurancyclopentane] skeleton units.

A Cu(acac)2-catalyzed cyclization reaction of 2-cyanobenzaldehydes with 2-isocyanoacetates has been successfully developed providing an efficient strategy for the synthesis of substituted 1-aminoisoquinolines. The reaction proceeds smoothly under mild conditions with high efficiency, and might provide an alternative strategy for the synthesis of 1-aminoisoquinoline containing molecules.

An Au(i)-catalyzed cyclization/1,2-rearrangement/aromatization cascade of 3-yne-1,2-diols has been successfully realized. This reaction not only provides a new and efficient strategy for the synthesis of substituted cycloalka[b]furan compounds as well as their derivatives, but might also facilitate related biological studies.

A Zn(OTf) 2 -promoted cyclization reaction of tosylhydrazones with 2-(dimethylamino)malononitrile has been successfully developed providing an efficient strategy for the synthesis of substituted 1-tosyl-1H-pyrazoles.

Aim: Cyclovirobuxinum D (CVB-D), an alkaloid isolated from the Chinese medicinal plant Buxus microphylla, has been found to be effective to treat cardiac insufficiency, arrhythmias and coronary heart disease. In the present study, we investigated the effects of CVB-D on the inflammatory responses in lipopolysaccharide (LPS)-stimulated murine macrophages in vitro and the underlying mechanisms., Methods: Murine macrophage cell line RAW264.7 cells were incubated in the presence of LPS (0.1 μg/mL) for 24 h. The cell viability was measured using MTT assay. The release of NO and cytokines were detected using the Griess test and ELISA, respectively. The mRNA and protein levels were determined using RT-PCR and Western blot, respectively. Reporter gene assays were used to analyze the transcriptional activity of NF-κB., Results: Treatment of RAW264.7 cells with CVB-D (25-300 μmol/L) did not affect the cell viability. Pretreatment with CVB-D (50, 100 and 200 μmol/L) concentration-dependently decreased NO release and iNOS expression in LPS-treated RAW264.7 cells (its IC50 value in inhibition of NO production was 144 μmol/L). CVB-D also concentration-dependently inhibited the secretion and mRNA expression of IL-1β and IL-6 in LPS-treated RAW264.7 cells. Furthermore, CVB-D remarkably inhibited the phosphorylation of STAT1 and STAT3, as well as JAK2 in LPS-treated RAW264.7 cells, but did not affect the activation of NF-κB and MAPKs pathways. Pretreatment with the JAK2 specific inhibitor AG490 (30 μmol/L) produced similar effects on NO release and iNOS expression in LPS-treated RAW264.7 cells., Conclusion: CVB-D exerts anti-inflammatory effects in LPS-stimulated murine macrophages in vitro at least in part by blocking the JAK-STAT signaling pathway. The anti-inflammatory actions of CVB-D may contribute to its cardioprotection.

Aim: Sinomenine (SIN) is an alkaloid found in the roots and stems of Sinomenium acutum, which has been used to treat rheumatic arthritis in China and Japan. In this study we investigated the effects of SIN on osteoclast survival in vitro and the mechanisms of the actions., Methods: Mature osteoclasts were differentiated from murine monocyte/macrophage cell line RAW264.7 through incubation in the presence of receptor activator of NF-κB ligand (RANKL, 100 ng/mL) for 4 d. The cell viability was detected using the CCK-8 method. The survival and actin ring construction of the osteoclasts were scored using TRACP staining and phalloidin-FITC staining, respectively. The apoptosis of the osteoclasts was detected by DNA fragmentation and Hoechst 33258 staining, and the cell necrosis was indicated by LDH activity. The activation of caspase-3 in osteoclasts was measured using Western blotting and the caspase-3 activity colorimetric method., Results: SIN (0.25-2 mmol/L) inhibited the viability of mature osteoclasts in dose-dependent and time-dependent manners, but did not affect that of RAW264.7 cells. Consistently, SIN dose-dependently suppressed the survival of mature osteoclasts. The formation of actin ring, a marker associated with actively resorbing osteoclasts, was also impaired by the alkaloid. SIN (0.5 mmol/L) induced the apoptosis of mature osteoclasts, which was significantly attenuated in the presence of the caspase-3 inhibitor Ac-DEVD-CHO. SIN increased the cleavage of caspase-3 in mature osteoclasts in dose-dependent and time-dependent manners. Furthermore, SIN dose-dependently enhanced caspase-3 activity, which was blocked in the presence of Ac-DEVD-CHO., Conclusion: Sinomenine inhibits osteoclast survival in vitro through caspase-3-mediated apoptosis, thus it is a potential agent for treating excessive bone resorption diseases.

Background: The recent H1N1 pandemic provided an opportunity to conceptually assess the possibility of rapidly providing a "hyperimmune" human immunoglobulin (H-IVIG) to an emerging infectious disease, in useful quantities with respect to public health. Commercial-scale H-IVIG production from plasma collected from donors convalescent from or vaccinated against pandemic influenza A (H1N1) virus is described., Study Design and Methods: A special protocol was implemented for the collection, processing, and shipment of plasma from previously qualified source plasma donors, self-identifying as convalescent from or vaccinated against H1N1 influenza. A licensed IVIG manufacturing process was utilized for the preparation of two commercial lots of approximately 50 kg 10% human IVIG preparation in total. The H1N1 hemagglutination inhibition and neutralization antibody titers of the resulting H-IVIG preparations were determined and compared with standard preparations., Results: Twenty-six plasma collection centers participated in the protocol. Donor enrollment exceeded 300 donors per week and within 30 days of protocol deployment plasma was being collected at a rate of more than 2000 L/week. Manufacture of both H-IVIG lots was unremarkable and both lots met the requirements for commercial release and the bulk of the product was distributed in normal commercial channels. Examination of plasma pools and final IVIG product confirmed pandemic H1N1 antibody titers substantially higher than those collected before the emergence of the pandemic H1N1 virus., Conclusions: This work demonstrates the feasibility of producing a H-IVIG preparation at large scale relatively rapidly, with a significant enrichment in antibodies to the H1N1 influenza, achieved by donor self-identification., (© 2011 American Association of Blood Banks.)

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.

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 investigate the antitumor effects of koumine in mice bearing H22 solid tumor and its effect on the immune system of the mice., Methods: The changes in spleen and tumor weights and blood cell count were observed after koumine treatment in BALB/c athymic mice bearing H22 solid tumor, using normal saline solution and 5-Fu as the controls., Results: Koumine significantly inhibited the tumor growth in a dose-dependent manner. The spleen index and blood cell counts in koumine group showed no significant differences from those in the saline control group, but higher than those in 5-Fu group., Conclusion: Koumine can significantly inhibit the growth of H22 solid tumor without obvious inhibitory effect on the immune system in 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 compare and evaluate the biocompatibility of three kinds of dentin bonding agents Xeno III (XO), Adper Prompt (AP), Single bond2 (SB) through cell culture in vitro., Methods: Three kinds of dentin bonding agents (XO, AP, SB) were applied on the surface of the dental slices which were 5.0 mm in diameter and 0.5 mm in depth. By immersing the slices into the DMEM culture medium, the maceration extracts were obtained. Normal dental pulps of teenagers were collected and human pulp fibroblast was cultured using tissue explant method. The fifth generation pulp cells were exposed to culture medium containing different concentrations of maceration extracts (100.0%, 50.0%, 25.0%, 12.5%) for 24, 72, 120 h. At last, MTT method was used to evaluate the cytotoxicity of the dentin bonding agents on human pulp fibroblast., Results: The results showed that all three kinds of dentin bonding systems had cytotoxicity to human pulp fibroblast in different degree in vitro. The cytotoxicity of XO and AP was less than SB. The difference was statistically significant (P<0.05)., Conclusion: The results of cell culture in vitro indicated that total-etching adhesives system has more irritation to pulp than self-etching adhesives system.

Objective: To explore the dynamic changes of blood sugar and body's signs in streptozotocin diabetic animal models., Methods: Rat and mouse diabetic models were established by a single intraperitoneal (ip) injection and 5-day successive ip injections of streptozotocin respectively. Blood sugar levels were measured. The food consumption index (consumption of food/body weight) and the water consumption index (consumption of water/body weight) were calculated., Results: Sixty five point zero percent male rats received streptozotocin, 60 mg/kg ip, developed diabetes mellitus. The blood sugar remained in high level between the 15th day and the 25th day after injection, and it began to decline afterwards. By 5-day ip injections of streptozotocin, 40 mg/kg daily, 90.0% male mice developed diabetes mellitus. Dynamic changes of blood sugar of diabetic mouse were similar to those of rats, except that the blood sugar of mice did not decline as obvious as that of rats. The changes of water consumption index were in best fit with the changes of blood sugar in both models, with correlation index r>0.970., Conclusion: The blood sugar of diabetic animal model stayed in high level from the 15th day to the 25th day after the beginning of injection. And the period is suitable for observing effect of anti-diabetic drugs. The water consumption index can reflect the blood sugar levels of diabetes animals.

Objective: To investigate the changes of serum inteferon-gamma (IFN-gamma) in mice bearing S-180 tumor and explore the role of the endogenous IFN-gamma in confining the transplanted tumor by intervention with immunomodulators., Methods: Mouse models bearing S-180 solid tumor were established and subjected to intragastric administration of Ganoderma lucidum polysaccharides (GLP) or cyclosporine A (CsA) at different daily doses for 9 consecutive days. Serum IFN-gamma levels were measured in untreated tumor-bearing mice and in those after completion of GLP or CsA treatments by enzyme-linked immunosorbent assay (ELISA), and the changes of the tumor weight in the treated mice were evaluated., Results: It was found for the first time that serum IFN-gamma levels in the tumor-bearing mice increased progressively within the initial 20 days after tumor implantation. The serum IFN-gamma levels in the 3 GLP-treated groups (at daily doses of 400, 200, and 100 mg/kg) all increased, which was the most obvious in 400 mg/kg GLP-treated group, and the tumor weight decreased significantly in response to GLP treatment, but the most conspicuous effect occurred with the daily dose of 200 mg/kg, and no significant statistical correlation was found between the two parameters. CsA treatment (at 20, 10, and 5 mg/kg, respectively) resulted in reduced serum IFN-gamma levels but produced virtually no effect on the tumor weight, and no obvious correlation was found between serum IFN-gamma level and the tumor weight., Conclusion: Increased serum IFN-gamma levels following GLP treatment are not significantly correlated to tumor growth inhibition in mice, and CsA reduces serum IFN-gamma levels without affecting the tumor weight, suggesting that endogenous IFN-gamma is not a major immunomodulating factor in growth inhibition of transplanted S-180 tumor.

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 effect of Ganoderma lucidum polysaccharides (GLP) on the nucleotide contents and cell cycle distribution of the tumor cells in S180 ascitic tumor-bearing mice and explore the possible mechanism of the antitumor effect of GLP., Methods: Mice bearing S180 ascitic tumor were subjected to intragastric administration of GLP (100, 200, and 400 mg/kg), normal saline or subcutaneous injection of cyclophosphamide (CTX) at 25 mg/kg, respectively. The treatment was given once daily for 9 consecutive days, after which the ascitic tumor cells were harvested for determination of the RNA and DNA contents and their ratio as well as the cell cycle alterations. Laser scanning confocal microscopy and acridine orange staining was performed to evaluate the DNA and RNA fluorescence intensity, and flow cytometry with propidium iodide (PI) staining was utilized for cell cycle analysis of the tumor cells., Results: Compared with normal saline group, the tumor cells in the 3 GLP groups all showed reduced RNA and DNA contents, and this reduction was statistically significant in 200 mg/kg GLP group (P=0.000). Significantly reduced RNA/DNA ratio was noted in all the 3 GLP groups (P=0.003, 0.000, 0.008 corresponding to 400, 200, and 100 mg/kg groups), suggesting that ganoderma polysaccharides more effectively reduced RNA content than DNA content. CTX also resulted in reduced RNA and DNA contents but not the RNA/DNA ratio. At the doses of 400, 200, and 100 mg/kg, GLP increased the percentage of G2/G2 phase cells (P=0.003, 0.000, and 0.000) whereas CTX showed the contrary effect (P=0.000). GLP produced no obvious effect on S-phage cells but CTX significantly reduced their percentage (P=0.000). GLP at the 3 doses all decreased the percentage of G2/M phase tumor cells (P=0.014, 0.049, 0.016) and CTX again induced contrary effect (P=0.000)., Conclusion: With different effects from CTX on DNA and RNA contents and cell cycle, GLP inhibits DNA and RNA synthesis in the tumor cells by mobilizing the host immune function to interfere with the normal cell cycles, which might be one of the mechanisms for the antitumor effect of GLP.

Objective: To study the antitumor effect of saponin extracted from Tupistra chinensis Baker (STCB) against mouse sarcoma S-180 cell proliferation in vitro and in vivo and explore the primary mechanism of this effect., Methods: Cytotoxic effect of STCB on S-180 cells in vitro was evaluated by MTT colorimetry, and its effect against in vitro tumor growth was tested in Kunmin mice bearing S-180 implanted tumor. The morphological and ultrastructural changes of S-180 cells after saponin treatment in vitro were examined with light and transmission electron microscope. Flow cytometry was performed to examine the cell cycle and apoptosis of S180 cells treated with different concentrations of STCB with propidium iodide staining., Results: STCB could markedly inhibit S-180 cell proliferation in vitro with 50% inhibitory concentration of 34.64 microg/ml. STCB given by intragastric administration also significantly inhibited the growth of S-180 solid tumor, and the inhibition rate exceeded 30% at the dose of 0.5 g/kg, reaching 54.86% at 2 g/kg. Electron microscopy and flow cytometry revealed increased S180 tumor cell apoptotic rate with the increment of saponin concentration, along with increased percentage of cells in S phase and decreased cells in G(2)/M phase in response to 10 or 30 microg/ml STCB treatment. At the concentration of 60 microg/ml, however, STCB resulted in an opposite effect on the cell cycles, presumably due to its interference with mitosis at high concentrations., Conclusions: STCB inhibits the growth of S-180 cells both in vivo and in vitro possibly by inducing cell apoptosis and interfering with the cell cycle progression of the tumor cells.

Objective: To determine if Ganoderma polysaccharides can antagonize prostaglandin E2 (PGE2)-induced suppression of murine splenocyte interferongamma (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) mRNA expression., Methods: Mixed lymphocyte culture reaction was used as the experimental model. The expressions levels of IFN-gamma and TNF-alpha mRNA were measured by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR)., Results: After the cultures were treated with PGE2 for 4 h, IFN-gamma mRNA expression was reduced as compared with the control, which was especially obvious when PGE2 concentrations exceeded 10 micromol/L (P<0.01). Ganoderma polysaccharides above 100 mg/L showed partial antagonistic effect against the inhibition of IFN-gamma by PGE2 at the fixed concentration of 20 micromol/L. Further studies indicated that PGE2 (20 micromol/L) impaired the expression of TNF-alpha mRNA after an 8-hour incubation and Ganoderma polysaccharides above 100 mg/L could partially antagonize this effect., Conclusion: Ganoderma polysaccharides can partially antagonize PGE2-induced suppression of murine splenocyte IFN-gamma and TNF-alpha mRNA expression.

Objective: To observe the effects of cyclosporin A (CsA) on gene expression profiles of NIT-1 pancreatic beta cells cell line using microarray technique., Methods: NIT-1 cells were exposed to cyclosporin A treatment (10 micromol/L) for 24 h and the differential gene expressions were assessed using microarray technique., Results: After 24h of CsA treatment, 38 of the 4096 genes tested were up-regulated, including 13 genes with known functions involving stress response, cell growth and protein synthesis. Meanwhile 46 genes were down-regulated, including 25 genes with known functions involving cell growth and maturation, oxidative phosphorylation and protein synthesis. Changes of gene expression of Zfr,Tpi and Pax6 were confirmed by semi-quantitative reverse transcription polymerase chain reaction., Conclusions: CsA treatment for 24 h induces changes in the gene expression profiles of NIT-1 pancreatic beta cells. Down-regulation of the genes related to cell growth and maturation, oxidative phosphorylation and protein synthesis may partly explain the mechanisms that CsA inhibits the release of insulin from pancreatic beta cells.

Objective: To isolate and purify a new polysaccharid Streptomyces polysaccharid polysaccharide (SMP ), from cultured broth of a Streptomyces sp.strain and perform its structural analysis., Methods: Ethanol was used to precipitate the polysaccharides and macromolecules from the broth. The proteins in the precipitate were removed by Sevage method and purification of SMP was carried out by DEAE-celluloseion-exchange chromatography and Sephadex G-25 gel filtration. The chemical structure of the SMP was determined by combined application of high performance liquid chromatography (HPLC), UV, IR and 1H-NMR spectroscopy, supplemented by periodate oxidation and Smith degradation., Results: The purified SMP was neutral by a relative molecular mass of approximately 4 855. Sugar analysis showed that SMP contained glucose and fructose residues in an approximate molar ratio of 22:1 (10.96 to 0.48). The glycosidic linkages were estimated to be (1-6)- alpha-D- pyranoside form., Conclusion: SMP is characterized as a (1-6)- alpha-D- pyranose.

Objective: To investigate the effects of cyclosporine A on the proliferation and pol alpha1 mRNA expression of cultured NIT-1 beta cells., Methods: After exposure to cyclosporine A at various concentrations (0.05 to 10 micromol/L) for 48 h and 72 h, NIT-1 cell proliferation was analyzed by MTT assay and the gene expression determined by reverse transcriptional PCR (RT-PCR)., Results: Forty-eight-hour and 72-hour cyclosporine A exposure inhibited the cell proliferation in a concentration- dependent manner, and at the concentration of 10 micromol/L, cyclosporine A also decreased pol alpha1 mRNA expression after a 48-hour exposure., Conclusion: Cyclosporine A can effectively inhibit the proliferation of NIT-1 cells possibly through down-regulating the expression of pol alpha1 mRNA.

Objective: To determine the effect of koumine parenteral solution on the nervous, respiratory and cardiovascular systems of experimental animals., Methods: Mouse spontaneous activities under the influence of the koumine injection were recorded with a photoelectric counter, and canine femoral artery pressure was determined by CYS-0.5 pressure transducer, respiratory curve described with TB-611 tension transducer and electrocardiogram (ECG) recorded with subcutaneous electrodes in the extremities after the injection. The above indices were automatically sampled and processed by multifunctional signal processor after being inputt to a computer. In this experiment, we observed the changes in the general behavior of the mice and their spontaneous activities within 15 min, along with the heart rate, maximum, minimum, and mean value of cardiac electric voltage, mean arterial pressure, respiratory rate and respiratory depth of the dogs before and at 10, 20, 30, 60, 90, 120 min after koumine injection., Results: Koumine injection significantly decreased mouse spontaneous activities in moderate and high-dose groups, but did not produce obvious effect on the respiratory system, mean arterial pressure, and maximum, minimum, and mean values of cardiac electric voltage in dogs. The heart rate of the dogs did not undergo obvious changes in response to the injection at a low dose, but median and high doses of the injection produced obvious effects., Conclusion: Koumine injection has definite sedative effect in mice, and does not affect the respiratory and cardiovascular systems of dogs with the exception of the heart rate.

Objective: To determine the effects of chronic valproic acid sodium (VPA) treatment and subsequent withdrawal on the release of glutamate (Glu) and glutamine (Gln) by C6 glioma cells, so as to understand the role of Glu and Gln released by astrocytes in the antiepileptic mechanism of VPA and the rebound mechanism of VPA withdrawal., Methods: C6 glioma cells were maintained for 2 weeks in DMEM medium containing 50 mg/L VPA to establish the cell model of chronic VPA treatment. High-performance liquid chromatography (HPLC) was performed to detect the levels of Glu and Gln released by C6 glioma cells after chronic VPA treatment and subsequent withdrawal., Results: Chronic VPA treatment increased Glu release of C6 glioma cells, and subsequent VPA withdrawal resulted in sustained decrease in the Glu level, reaching the lowest level 12 h after VPA withdrawal, which was obviously lower than the control level. Gradual but significant increase of Glu level was then observed to approach the control level till 48 h after VPA withdrawal. For Gln release, chronic VPA treatment resulted in its decrease while after VPA withdrawal, gradual increase occurred to recover the control level., Conclusions: Chronic VPA treatment can increase Glu and decrease Gln release by C6 glioma cells. Glu level undergoes a rebound in response to VPA withdrawal, while the relatively even changes of Gln level in the same setting may not involve VPA withdrawal rebound mechanism.

Objective: To examine the effects of sodium valproate(VPA) treatment and withdrawal on the expression of GABA transporter-3 (GAT-3) and GABA transaminase (GABA-T) mRNA in C6 glioma cells, and to explore the role of GAT-3 and GABA-T in the rebound mechanism of VPA withdrawal., Methods: C6 glioma cells were maintained for 2 weeks in DMEM medium containing VPA (50 mg/L) to establish the cell model of chronic exposure to VPA. Semi-quantitative RT-PCR was used to examine the changes of GAT-3 and GABA-T mRNA expression in response to VPA treatment and withdrawal., Results: Chronic exposure to VPA down-regulated GAT-3 mRNA expression to 39.1% +/-0.5% from 46% +/-1.3% in the control group; After VPA withdrawal, GAT-3 mRNA expression level kept decreasing, reaching the minimum (11.7% +/-1.6%) 24 h after the withdrawal and with an increase to the level of 33.5%+/-1.1% after another 24 hr. GABA-T mRNA expression was up-regulated to 71.31% +/-8.39% from 34.77% +/-2.36% of the control level after VPA treatment, the withdrawal of which resulted in decreased GABA-T mRNA expression. Till 12 h after the withdrawal, the GABA-T mRNA expression level decreased to the minimum, 25.36% +/-7.68%., Conclusions: Chronic treatment with VPA can down-regulate GAT-3 mRNA expression and up-regulate GABA-T mRNA expression in C6 glioma cells, and this undulation may involve VPA withdrawal rebound.

Objective: To study the apoptosis-inducing effect of koumine on human colon adenocarcinoma LoVo cells in vitro., Methods: After koumine (50 mmol/L) treatment in vitro, the LoVo cells were examined under light microscope, transmission electron microscope and fluoroscope respectively for apoptosis, and the cell cycle distribution was analyzed using flow cytometry., Results: The percentage of apoptotic cells increased in a time-dependent manner after the cells were treated with koumine, whose action exhibited remarkable cell cycle specificity. The percentage of LoVo cells in G(0)/G(1) phase rose from 31.3% to 42.3% and the percentage of cells in S phase fells from 62.0% to 38.7%., Conclusion: Koumine can induce apoptosis of LoVo cells in a time-dependent manner and inhibit the DNA synthesis in LoVo cells, thereby blocking the cell cycle from G1 to S phase.

Objective: To investigate the effects of cyclosporin A on insulin release and the gene expression profiles of mitochondrial oxidative phosphorylation in NIT-1 cells., Methods: NIT-1 cells were exposed to cyclosporin A (10 micromol/L) for 24 and 48 h respectively, after which the amount of insulin release was determined by means of radioimmunoassay (RIA), and the differential expressions of Nuox23, Cox7c and Atp5K genes assessed by semi-quantitative reverse transcription polymerase chain reaction., Results: Cyclosporin A reduced insulin release in the cell culture after 24 and 48 h exposure and decreased Nuox23, Cox7c and Atp5K mRNA expressions., Conclusion: Cyclosporin A induces inhibition of insulin release in NIT-1 cells, possibly due to the reduction of ATP synthesis involving the down-regulation of the gene expression of mitochondrial oxidative phosphorylation enzymes.

Objective: Investigating the effect of GLB7 on cAMP in murine peritoneal macrophages to provide a scientific evidence for the immunomodulatory mechanism., Method: Cell culture and radio-immunological assay of cAMP were used., Result: GLB7 increased the production of cAMP in a concentration and time dependent manner in murine peritoneal macrophages., Conclusion: The immunopotentiating effect of GLB7 may be due to the activation of macrophages that leads to the increase of cAMP.

Aim: To study the effects of gypenosides (Gyp) on lymphocyte transformation and DNA polymerase II activity., Methods: Lymphocyte transformation response was induced by concanavalin A and lipopolysaccharides respectively. The activity of DNA polymerase II and DNA synthesis were assayed with TTP and [3H]TdR incorporation respectively in mixed lymphocyte culture test., Results: Gyp 2.5-20 mg L-1 enhanced splenic T- and B- cell transformation, increased the DNA synthesis and potentiated the activity of DNA polymerase II. However, Gyp > 40 mg L-1 showed contrary effects., Conclusion: Gyp regulated lymphocyte transformation and DNA synthesis by regulating DNA polymerase II activity.

The activity of DNA polymerase alpha in splenocytes of 24-month-old mice was about 35.6% lower than that of 3-month-old mice. Aged mice were intraperitoneally administered Ganoderma polysaccharides (GL-B) once a day for 4 days and then the activity of the enzyme was assessed. The results showed that GL-B at doses of 25 and 50 mg/kg-1 enhanced the activity of the enzyme in aged mouse splenocytes by 44.0 and 58.8% respectively. In addition, the mixed lymphocyte response to alloantigen, automatic proliferation and IL-2 production of splenocytes in aged mice declined as compared with that in young adult mice. GL-B (50, 100, 200 micrograms.ml-1) was found to restore those parameters to the levels of that of young mice in vitro.

Mixed lymphocyte response was used as a main model through all the experiments In a series of concentrations (25, 50, 100, and 200 micrograms/ml), Ganoderma polysaccharides (GL-B) promoted the production of interleukin 2 (IL-2) in a concentration-dependent manner after initiation of culture for 12 h and increased the total cell recovery as well as that of Lyt 2+ and L3T4+ cells after 4 days of culture. The data also show that the polysaccharides markedly enhanced the cytotoxicity of cytotoxic T lymphocytes, which was increased by 100% at the concentration of 200 micrograms/ml.