Growth and inhibition of zinc anode dendrites in Zn-air batteries:Model and experiment

Zinc(Zn)-air batteries are widely used in secondary battery research owing to their high theoretical energy density,good electrochemical reversibility,stable discharge performance,and low cost of the anode active material Zn.However,the Zn anode also leads to many challenges,including dendrite growth,deformation,and hydrogen precipitation self-corrosion.In this context,Zn dendrite growth has a greater impact on the cycle lives.In this dissertation,a dendrite growth model for a Zn-air battery was established based on electrochemical phase field theory,and the effects of the charging time,anisotropy strength,and electrolyte temperature on the morphology and growth height of Zn dendrites were studied.A series of experiments was designed with different gradient influencing factors in subsequent experiments to verify the theoretical simulations,including elevated electrolyte temperatures,flowing electrolytes,and pulsed charging.The simulation results show that the growth of Zn dendrites is controlled mainly by diffusion and mass transfer processes,whereas the electrolyte temperature,flow rate,and interfacial energy anisotropy intensity are the main factors.The experimental results show that an optimal electrolyte temperature of 343.15 K,an optimal electrolyte flow rate of 40 ml·min^(-1),and an effective pulse charging mode.

Engineering hydrophobic protective layers on zinc anodes for enhanced performance in aqueous zinc-ion batteries

Aqueous zinc-ion batteries possess substantial potential for energy storage applications;however,they are hampered by challenges such as dendrite formation and uncontrolled side reactions occurring at the zinc anode.In our investigation,we sought to mitigate these issues through the utilization of in situ zinc complex formation reactions to engineer hydrophobic protective layers on the zinc anode surface.These robust interfacial layers serve as effective barriers,isolating the zinc anode from the electrolyte and active water molecules and thereby preventing hydrogen evolution and the generation of undesirable byproducts.Additionally,the presence of numerous zincophilic sites within these protective layers facilitates uniform zinc deposition while concurrently inhibiting dendrite growth.Through comprehensive evaluation of functional anodes featuring diverse functional groups and alkyl chain lengths,we meticulously scrutinized the underlying mechanisms influencing performance variations.This analysis involved precise modulation of interfacial hydrophobicity,rapid Zn^(2+)ion transport,and ordered deposition of Zn^(2+)ions.Notably,the optimized anode,fabricated with octadecylphosphate(OPA),demonstrated exceptional performance characteristics.The Zn//Zn symmetric cell exhibited remarkable longevity,exceeding 4000 h under a current density of 2 mA cm^(-2)and a capacity density of 2 mA h cm^(-2),Furthermore,when integrated with a VOH cathode,the complete cell exhibited superior capacity retention compared to anodes modified with alternative organic molecules.

Inhibitory effect of arsenic trioxide on angiogenesis and expression of vascular endothelial growth factor in gastric cancer

AIM: To investigate the inhibitory effect of As2O3 on angiogenesis of tumor and expression of vascular endothelial growth factor (VEGF) in tumor cells in vivo and in vitro. METHODS: The solid tumor model was formed in nude mice with the gastric cancer cell line SGC-7901. The animals were randomly divided into three groups. As2O3 was injected into the arsenic-treated groups (2.5 mg/kg and 5 mg/kg) and the same volume of saline solution was injected into the control group. Microvessel density (MVD) and expression of VEGF were detected with immunofluorescence laser confocal technology. Further expression of VEGF protein and VEGF mRNA was measured with Western bloting and fluorescence quantitative RT- PCR in SGC-7901 cells treated with As2O3. RESULTS: In nude mice, after treatment with 5 mg/kg and 2.5 mg/kg As2O3 respectively, about 50% and 30% tumor growth inhibition were observed correspondingly (P < 0.05, P < 0.05). Decrease in MVD appeared in As2O3-treated tumors compared with control group (P < 0.001, P < 0.001). MVD in tumors was significantly lower in 5 mg/kg group than in 2.5 mg/kg group (P < 0.01). The fluorescence intensity levels of VEGF in tumor cells were significantly lowered in the arsenic-treated groups (P < 0.01, P < 0.01). The fluorescence intensity level of VEGF in 5 mg/kg group was lower than that in 2.5 mg/ kg group (P < 0.01). In vitro, the expression of VEGF protein decreased in dose- and time-dependent manner after the treatment with As2O3, but in VEGF mRNA no significant difference was found between the control group and the treated groups. CONCLUSION: As2O3 can inhibit solid tumor growth by inhibiting the formation of new blood vessels. One of the mechanisms is that As2O3 can inhibit VEGF protein expression.

Effect of arsenic trioxide on vascular endothelial cell proliferation and expression of vascular endothelial growth factor receptors Flt-1 and KDR in gastric cancer in nude mice

AIM： To investigate the effect of arsenic trioxide （As2O3） on expression of vascular endothelial growth factor receptor-1 （VEGFR-1, Flt-1） and VEGFR-2 （KDR） in human gastric tumor cells and proliferation of vascular endothelial cells.METHODS： The solid tumor model was formed in nude mice with the gastric cancer cell line SGC-7901. The animals were treated with As2O3. Microvessel density （MVD） and expression of Flt-1 and KDR were detected by immunofluorescence laser confocal microscopy. SGC-7901 cells were treated respectively by exogenous recombinant human VEGF165 or VEGF165 ＋ As2O3. Cell viability was measured by MTT assay. Cell viability of ECV304 cells was measured by MTT assay, and cell cycle and apoptosis were analyzed using flow cytometry.RESULTS： The tumor growth inhibition was 30.33% and 50.85%, respectively, in mice treated with As2O3 2.5 and 5 mg/kg. MVD was significantly lower in arsenic-treated mice than in the control group. The fluorescence intensity levels of Flt-1 and KDR were significantly less in the arsenic-treated mice than in the control group. VEGF165 may accelerate growth of SGC7901 cells, but As2O3 may disturb the stimulating effect of VEGF165. ECV304 cell growth was suppressed by 76.51%, 71.09% and 61.49% after 48 h treatment with As2O3 at 0.5, 2.5 and 5 μmol/L, respectively. Early apoptosis in the As2O3- treated mice was 2.88-5.1 times higher than that in the controls, and late apoptosis was 1.17-1.67 times higher than that in the controls.CONCLUSION： Our results showed that As2O3 delays tumor growth, inhibits MVD, down-regulates Flt-1 and KDR expression, and disturbs the stimulating effect of VEGF165 on the growth of SGC7901 cells. These results suggest that As2O3 might delay growth of gastric tumors through inhibiting the paracrine and autocrine pathways of VEGF/VEGFRs.

Sirolimus inhibits growth of human hepatoma cells alone or combined with tacrolimus, while tacrolimus promotes cell growth

AIM: Standard immunosuppression after organ transplantation stimulates tumor growth. Sirolimus has a strong antiproliferative and a tumor inhibiting effect. The purpose is to assess the effect on tumor growth of the immunosuppressive compounds sirolimus and tacrolimus alone and in combination on cells of human hepatocellular carcinoma.METHODS: We used the human cell lines SK-Hep 1 and Hep 3B derived from hepatocellular carcinoma. Proliferation analyses after treatment with sirolimus, tacrolimus, or the combination of both were performed. FACS analyses were done to reveal cell cycle changes and apoptotic cell death. The expression of apoptosis-related proteins was estimated by Western blots.RESULTS: Sirolimus alone or combined with tacrolimus inhibited the growth of both cell lines after 5 d by up to 35% in SK-Hep 1 cells, and by up to 68% in Hep 3B cells at 25 ng/mL. Tacrolimus alone stimulated the growth by 12% after 5 ng/mL and by 25% after 25 ng/mL in Hep 3B cells. We found an increase of apoptotic Hep 3B cells from 6 to 16%, and a G1-arrest in SK-Hep 1 cells with an increase of cells from 61 to 82%, when sirolimus and tacrolimus were combined. Bcl-2 was down-regulated in Hep 3B, but not in SK-Hep 1 cells after combined treatment.CONCLUSION: Sirolimus appears to inhibit the growth of hepatocellular carcinoma cells alone and in combination with tacrolimus. Sirolimus seems to inhibit the growth stimulation of tacrolimus.

Growth inhibition of high-intensity focused ultrasound on hepatic cancer in vivo

AIM： To investigate the damaging effect of high-intensity focused ultrasound （HIFU） on cancer cells and the inhibitory effect on tumor growth. METHODS： Hurine H22 hepatic cancer cells were treated with HIFU at the same intensity for different lengths of time and at different intensities for the same length oftime in vitro, the dead cancer cells were determined by trypan blue staining. Two groups of cancer cells treated with HIFU at the lowest and highest intensity were inoculated into mice. Tumor masses were removed and weighed after 2 wk, tumor growth in each group was confirmed pathologically.RESULTS： The death rate of cancer cells treated with HIFU at 1 000 W/cm^2 for 0.5, 1, 2, 4, 8, and 12 s was 3.11±1.21%, 13.37±2.56%, 38.84±3.68%, 47.22±5.76%,87.55±7.32%, and 94.33±8.11%, respectively. A positive relationship between the death rates of cancer cells and the length of HIFU treatment time was found （r = 0.96,P〈0.01）. The death rate of cancer cells treated with HIFU at the intensity of 100, 200, 400, 600, 800, and 1 000 W/cm^2 for 8 s was 26.31±3.26%, 31.00±3.87%, 41.97±5.86%,72.23±8.12%, 94.90±8.67%, and 99.30±9.18%, respectively. A positive relationship between the death rates of cancer cells and the intensities of HIFU treatment was confirmed （r= 0.98, P〈0.01）. The cancer cells treated with HIFU at 1 000 W/cm^2 for 8 s were inoculated intomice ed into. The tumor inhibitory rate was 90.35% compared to the control （P〈0.01）. In the experimental group inoculated with the cancer cells treated with HIFU at 1 000 W/cm^2 for 0.5 s, the tumor inhibitory rate was 22.9% （P〈0.01）. By pathological examination, tumor growth was confirmed in 8 out of 14 mice （57.14%, 8/14） inoculated with the cancer cells treated with HIFU at 1 000 W/cm^2 for 8 s, which was significantly lower than that in the control （100%, 15/15, P〈O.05）.CONCLUSION： HIFU is effective on killing or damage of H22 hepatic cancer cells in vitro and on inhibiting tumor growth in mice ex vivo.

The Arabidopsis P450 protein CYP82C2 modulates jasmonateinduced root growth inhibition, defense gene expression and indole glucosinolate biosynthesis

Jasmonic acid （JA） is a fatty acid-derived signaling molecule that regulates a broad range of plant defense responses against herbivores and some microbial pathogens. Molecular genetic studies have established that JA also performs a critical role in several aspects of plant development. Here, we describe the characterization of the Arabidopsis mutantjasmonic acid-hypersensitivel-1 （jah1-1）, which is defective in several aspects of JA responses. Although the mutant exhibits increased sensitivity to JA in root growth inhibition, it shows decreased expression of JA-inducible defense genes and reduced resistance to the necrotrophic fungus Botrytis cinerea. Gene cloning studies indicate that these defects are caused by a mutation in the cytochrome P450 protein CYP82C2. We provide evidence showing that the compromised resistance of thejah1-1 mutant to B. cinerea is accompanied by decreased expression of JA-induced defense genes and reduced accumulation of JA-induced indole glucosinolates （IGs）. Conversely, the enhanced resistance to B. cinerea in CYP82C2-overexpressing plants is accompanied by increased expression of JA-induced defense genes and elevated levels of JA-induced IGs. We demonstrate that CYP82C2 affects JA-induced accumulation of the IG biosynthetic precursor tryptophan （Trp）, but not the JA-induced IAA or pathogen-induced camalexin. Together, our results support a hypothesis that CYP82C2 may act in the metabolism of Trp-derived secondary metabolites under conditions in which JA levels are elevated. Thejah1-1 mutant should thus be important in future studies toward understanding the mechanisms underlying the complexity of JA-mediated differential responses, which are important for plants to adapt their growth to the ever-changing environments.

Growth and physiological responses of Agriophyllum squarrosum to sand burial stress

Agriophyllum squarrosum is an annual desert plant widely distributed on mobile and semi-mobile dunes in all the sandy deserts of China. We studied the growth and physiological properties of A. squarrosum seedlings under different sand burial depths in 2010 and 2011 at Horqin Sandy Land, Inner Mongolia to understand the ability and mechanism that A. squarrosum withstands sand burial. The results showed that A. squarrosum had a strong ability to withstand sand burial. Its survival rate, plant height and biomass increased significantly at a burial depth 25% of seedling height and decreased significantly only when the burial depth exceeded the height of the seedlings; some plants still survived even if the burial depth reached 266% of a seedling height. The malondialdehyde （MDA） content and membrane permeability of the plant did not change significantly as long as the burial depth was not greater than the seedling height; lipid peroxidation increased and cell membranes were damaged if the burial depth was increased further. When subjected to sand burial stress, superoxide dismutase （SOD） and peroxidase （POD） activities and free proline content increased in the seedlings, while the catalase （CAT） activity and soluble sugar content decreased. Sand burial did not lead to water stress. Reductions in photosynthetic area and cell membrane damage caused by sand burial may be the major mechanisms increasing mortality and inhibiting growth of the seedling. But the increases in SOD and POD activities and proline content must play a certain role in reducing sand burial damage.

Inhibition of human gastric carcinoma cell growth by atofluding derivative N_3-o-toluyl-fluorouracil

AIM： To evaluate the growth inhibition efficacy of atofluding derivative N3-o-toluyl-fluorouracil （TFU） on human gastric carcinoma cell lines SGC-7901 and MKN-45. METHODS： Cell growth inhibition by TFU was measured by MTT and clonogenic assays without or with liver microsomal enzymes. Xenografts of cancer cells in nude mice were employed to study the anti-proliferative effects of TFU in vivo. RESULTS： TFU inhibited the growth of SGC-7901 and MKN-45 cells. However, the inhibitory effects of TFU on cell growth were not significant. The inhibition rates were enhanced in the presence of liver microsomal enzymes, ranging 4.73%-48.57% in SGC-7901 cells and 9.0%-62.02% in MKN-45 cells. In v/vo, TFU delayed the growth of SGC-7901 and MKN-45 cells in nude mice. The inhibition rates were 40.49%, 63.24%, and 75.98% in SGC-7901 cells and 40.76%, 61.41%, and 82.07% in MKN-45 cells when the oral doses were 25, 50, and 100 mg/kg, respectively. TFU treatment was generally well tolerated by mice with less than 20% reduction in body weight. CONCLUSION： TFU inhibits the growth of human gastric carcinoma cells. The inhibition rates are increased in the presence of liver microsomal enzymes. The efficacy of TFU may be associated with the sustaining release of 5-fluorouracil （5-FU） mediated by the enzymes.

Inhibitory effects of extracellular adenosine triphosphate on growth of esophageal carcinoma cells

AIM： To study the growth inhibitory effects of ATP on TE-13 human squamous esophageal carcinoma cellsin vitro.METHODS： NTT assay was used to determine the inhibition of proliferation of ATP or adenosine （ADO） on TE-13 cell line. The morphological changes of TE-13 cells induced by ATP or ADO were observed under fluorescence light microscope by acridine orange （AO）/ethidium bromide （EB） double stained cells. The intemudeosomal fragmentation of genomic DNA was detected by agarose gel electrophoresis. The apoptotic rate and cell cycle after treatment with ATP or ADO were determined by flow cytometry.RESULTS： ATP and ADO produced inhibitory effects on TE-13 cells at the concentration between 0.01 and 1.0 mmol/L. The ICs0 of TE-13 cells exposed to ATP or ADO for 48 and 72 h was 0.71 or 1.05, and 0.21 or 0.19 mmol/L, respectively. The distribution of cell cycle phase and proliferation index （PI） value of TE-13 cells changed, when being exposed to ATP or ADO at the concentrations of 0.01, 0.1, and 1 mmol/L for 48 h. ATP and ADO inhibited the cell proliferation by changing the distribution of cell cycle phase via either G0/G1 phase （ATP or ADO, 1 mmol/L） or S phase （ATP, 0.1 mmol/L） arrest. Under light microscope, the tumor cells exposed to 0.3 mmol/L ATP or ADO displayed morphological changes of apoptosis. A ladder-like pattern of DNA fragmentation was obtained from TE-13 cells treated with 0.1-1 mmol/L ATP or ADO in agarose gel electrophoresis. ATP and ADO induced apoptosis of TE-13 cells in a dose-dependent manner at the concentration between 0.03 and 1 mmol/L. The maximum apoptotic rate of TE-13 cells exposed to ATP or ADO for 48 h was 16.63% or 16.9%, respectively.CONCLUSION： ATP and ADO inhibit cell proliferation, arrest cell cycle, and induce apoptosis of TE-13 cell line.

Lentivirus mediated shRNA interference targeting MAT2B induces growth-inhibition and apoptosis in hepatocelluar carcinoma

AIM: To investigate the effects of lentivirus vector mediated short hairpin RNA interference targeting methionine adenosyltransferase 2β gene (LV-shMAT2B) on hepatocelluar carcinoma (HCC) cells. METHODS: We constructed four plasmids of RNA interference targeting the MAT2B gene. After LV-shMAT2B was transfected with L-02 cells and two kinds of HCC cells, cell viability and proliferation were measured with MTT and [3H]thymidine assays respectively. Flow cytometry was used to assess cell apoptosis. The level of S-adenosyl methionine (SAMe) in HepG2 cells was evaluated. The expressions of cyclin D1, cyclin D2, bcl-xL and bcl-xS were detected with western blot. RESULTS: We constructed LV-shMAT2B successfully. LV-shMAT2B was safe for human normal liver cells. LV-shMAT2B caused dramatic reduction in proliferation compared with controls in HCC cells Bel-7402 (P = 0.054) and HepG2 (P = 0.031). Flow cytometry analysis showed that cell apoptosis caused by LV-shMAT2B was greater in HCC cells Bel-7402 and HepG2 than in control induced by scrambled siRNA (P = 0.047), but apoptosis rates in L-02 induced by LV-shMAT2B and scrambled siRNA respectively had no significant difference. Moreover, LV-shMAT2B significantly suppressed expression of MAT2B leading to growth-inhibition effect on HCC cells by down-regulating cyclin D1. Apoptosis induced by LV-shMAT2B was involved indown-regulating bcl-xL and up-regulating bcl-xS. CONCLUSION: LV-shMAT2B can induce cell apoptosis and growth-inhibition in HCC cells. MAT2B may be a therapy target in HCC in the future.

Experimental Study on Inhibitory Effects of Diallyl Sulfide on Growth and Invasion of Human Osteosarcoma MG-63 Cells

The inhibitory effects of diallyl sulfide(DAS) derived from allicin on in vitro and in vivo proliferation of human osteosarcoma MG-63 cells and the action mechanism,and the influence of DAS on invasive capability of MG-63 cells were investigated in order to search for the novel medicines for osteosarcoma.In the in vitro experiment,MG-63 cells were treated with different concentrations of DSA,and the morphological changes of MG-63 cells were observed under an inverted phase microscope.MTT method was used to assay the proliferation of MG-63 cells.Semi-quantitative reverse transcription polymerase chain reaction(RT-PCR) was used to detect the VEGF mRNA expression level in MG-63 cells.By using Transwell invasion assay,the influence of DAS on invasive ability of MG-63 cells was tested.In the in vivo experiment,the nude mice MG-63 cells tumor-bearing model was established,and different concentrations of DAS were injected beside the tumor.Twenty-one days after treatment,the mice were killed,the tumor size and tumor inhibition rate were calculated.The microvessel density(MVD) was determined by using immunohistochemistry.In the in vitro experiment,different concentrations of DAS could obviously inhibit proliferation of MG-63 cells in a time-and concentration-dependent manner.RT-PCR revealed that the expression levels of VEGF mRNA in DSA groups(different concentrations) were significant reduced as compared with those in control group(all P<0.05).Transwell invasion assay indicated that in 20 and 40 μg/mL DAS groups,the number of migratory cells was 91.4±8.3 and 81.8±7.4 respectively,which was significantly declined as compared with that in control group(150.4±14.7,both P<0.05).In the in vivo experiment,DAS could significantly suppress the growth of MG-63 tumor-bearing tissue.Immunohistochemistry demonstrated that different concentrations(20 and 40 μg/mL) of DAS could significantly decrease MVD of MG-63 tumor-bearing tissue(all P<0.05).It was suggested that DAS could inhibit the growth of MG-63 cells probably by suppressing the expression of VEGF mRNA.

Transactivation of the TIEG1 confers growth inhibition of transforming growth factor-β-susceptible hepatocellular carcinoma cells

AIM:To investigate the role of transforming growth factor(TGF)-β-inducible early gene 1(TIEG1) in TGF-β-induced growth inhibition in hepatocellular carcinoma(HCC) cells.METHODS:Human hepatocyte and HCC cell lines with varied susceptibilities to TGF-β1 were tested by methylthiazoletetrazolium(MTT) assay.The expression changes of Smad2,Smad3,Smad4,Smad7,TIEG1 and TIEG2 gene following treatment with TGF-β1 in a TGF-β-sensitive hepatocyte cell line(MIHA),a TGF-β-sensitive hepatoma cell line(Hep3B) and two TGF-β-insensitive hepatoma cell lines(HepG2 and Bel7404) were examined.SiRNA targeting TIEG1 was transfected into Hep3B cells and the sensitivity of cells to TGF-β1 was examined.Overexpression of TIEG1 was induced by lentiviral-mediated transduction in TGF-β1-resistant hepatoma cell lines(Bel7404 and HepG2).MTT assay and 4',6-Diamidino-2-phenylindole staining were used to identify cell viability and apoptosis,respectively.The expression level of stathmin was measured by reverse transcriptase polymerase chain reaction and Western-blotting analysis,and stathmin promoter activity by TIEG1 was monitored by a luciferase reporter gene system.RESULTS:TIEG1 was significantly upregulated by TGF-β1 in the TGF-β1-sensitive HCC cell line,Hep3B,but not in the resistant cell lines.The suppression of TIEG1 by siRNAs decreased the sensitivity of Hep3B cells to TGF-β1,whereas the overexpression of TIEG1 mediated growth inhibition and apoptosis in TGF-β1-resistant HCC cell lines,which resembled those of TGF-β1-sensitive HCC cells treated with TGF-β1.Our data further suggested that stathmin was a direct target of TIEG1,as stathmin was signif icantly downregulated by TIEG1 overexpression,and stathmin promoter activity was inhibited by TIEG1 in a dose-dependent manner.CONCLUSION:Our data suggest that transactivation of TIEG1 conferred growth inhibition of TGF-β-susceptible human HCC cells.

Growth Inhibition and Apoptosis Induced by Retinoic Acid Combined with Interferon Alpha-2a on Transitional Cell Carcinoma of Bladder

Objective:To identify new favorable agents and develop novel approaches for the chemoprevention and treatment of superficial bladder cancer and investigate the effects of combination of retinoids and interferon α-2a on growth inhibition and apoptosis induction in bladder cancer cell lines. Methods:Four bladder cancer cell lines,grade 1 to 3,and two retinoids,all-trans-retinoic acid(ATRA),9-cis retinoic acid(9cRA),combined with interferon α-2a(INF),were used in the study.We compared the competence of these agents to inhibit growth,induce apoptosis,affect the expression of nuclear retinoid receptors,and modulate STAT1 protein. Results: Most of the bladder cancer cell lines were resistant to the effect of ATRA and 9cRA on growth inhibition and apoptosis induction,even at higher concentration(10 -5M).The effects of ATRA and 9c RA on cell growth and apoptosis were enhanced by INF α- 2a. Combination of ATRA and IFNα-2a induced RARβ and Stat 1 expression in three bladder cancer cell lines. Conclusion:The results demonstrated that INFα-2a synergize with the inhibitory effect of ATRA and 9c RA on the growth inhibition and apoptosis of bladder cancer cells in vitro,which suggested that it has a potential interest for the treatment of transitional cell carcinoma of bladder.

Growth Inhibition of Breast Cancer in Rat by AAV Mediated Angiostatin Gene

Objective： To observe growth inhibition effect of adeno-associated viral vectors （AAV） mediated angiostatin （ANG） gene on implanted breast cancer in rat and its mechanism. Methods： Gene transfer technique was used to transfer AAV-ANG to the tumor. Growth curves were drawn to observe the growth of breast cancer implanted in rat, and immunohistochemical method was used to detect the effects of angiostatin on microvesel density （MVD） of breast cancer implanted in rat. Results： Angiostatin inhibited the growth of breast cancer implanted in rat and decreased the microvessel density of tumor. Conclusion： Expression of an angiostatin transgene can suppress the growth of breast cancer implanted in rat through the inhibition of the growth of microvessels, surggesting that angiostatin gene transfer technique may be effective against breast cancer.

Growth-inhibitory effects of MOB2 on human hepatic carcinoma cell line SMMC-7721

AIM:To investigate the growth-inhibiting and apoptosis-inducing effects of the gene MOB2 on human hepatic carcinoma cell line SMMC-7721.METHODS:The full-length cDNA of the MOB2 gene was amplified from human umbilical vein endothelial cells.The correct full-length MOB2 cDNA was subcloned into the eukaryotic expression vector pEGFP-C1.After lipofection of the MOB2 gene into cancer cells,the levels of MOB2 protein in the cancer cells were detected by immunoblotting.To transfect the recombined plasmid vector pEGFP-CI-MOB2 into SMMC-7721 cells,the cells were cultured in Dulbecco's Modified Eagle'sMedium with 10% fetal calf serum and glutamine,and then mixed with liposomes,Lipofectamine 2000 and the plasmid vector pEGFP-CI-MOB2.RESULTS:We observed the growth and proliferation of SMMC-7721 cells containing pEGFP-CI-MOB2 and analyzed their apoptosis and growth cycle phases by flow cytometry.We successfully transfected the recombined plasmid vector pEGFP-CI-MOB2 into SMMC-7721 cells and screened for a single clone cell containing MOB2.After transfection,MOB2 enhanced growth suppression,induced apoptosis,increased the ratio of G0/G1,significantly inhibited the advance of cell cycle phase,and arrested cells in G0/G1 phase.CONCLUSION:MOB2 overexpression induces apoptosis and inhibits the growth of human hepatic cancer cells,which may be useful in gene therapy for hepatic carcinoma.

LOTUS, a potent blocker of Nogo receptor-1 causing inhibition of axonal growth

Glia-derived axonal growth inhibitory proteins limit functional repair following damage to the adult cen- tral nervous system （CNS）. Nogo proteins, myelin-as- sociated glycoprotein （MAG）, oligodendrocyte myelin glycoprotein （OMgp） and B lymphocyte stimulator （BLyS）, are 4 inhibitors that commonly interact with the neuronal receptor, Nogo receptor-1 （NgR1）, lead- ing to inhibition of axonal growth. Here, we demon- strate that lateral olfactory tract usher substance （LOTUS） binds to NgR1 and blocks the binding of all four ligands to NgR1, resulting in the suppression of axonal growth inhibition induced by these NgR1 li- gands. LOTUS allows neurons to overcome NgRl-me- diated axonal growth inhibition, raising the possibility that LOTUS may be useful in future therapeutic ap- proaches as an endogenous potent inhibitor of NgR1 for promoting neuronal regeneration.

Growth inhibition to three red tide microalgae by extracts of Ulva pertusa

Growth inhibition effect of different concentration of distilled water extract and four polar organic solvent (methanol, acetone, ether and chloroform) extracts of Ulva pertusa on three typical red tide microalgae (Heterosigma akashiwo, Alexandrium tamarense and Prorocentrum micans) were inves- tigated. Liquid-liquid fractionation and HPLC analysis for methanol extract of U. pertusa were carried out. Growth of the three microalgae was significantly inhibited by the distilled water extract of U. pertusa at relatively higher concentration. However, the cells of the three microalgae did not die completely even at high concentration. Methanol extract of U. pertusa showed the highest growth inhibition on the three mi- croalgae, and all the cells of the three microalgae were killed at relatively high concentration. The other three organic solvent extracts of U. pertusa had no apparent effect on the three microalgae. The results of bioassays and HPLC analysis suggested that the inhibitory substances in U. pertusa to the microalgal growth had relatively high polarities. H. akashiwo was the most sensitive one while A. tamarense was the most tolerant one to the growth inhibitory substances.

Inhibition of the growth and development of mosquito larvae of Culex quinquefasciatus(Diptera:Culicidae) treated with extract from leaves of Pseudocalymma alliaceum(Bignonaceae)

Objective:To determine larvicidal activity of the essential oil,hydrolat and botanical extracts derived from leaves ol Pseudocalymma alliaceum an mosquito larvae of Culex quinquefasciatus.Methods:Groups of twenty lanae were used in the larvicidal assays.The mortality,relative growth rate,the larval and pupal duration and viability was estimated.The essential oil was analyzed by solid phase microextraction using gas chromatography coupled to mass spectrometry.Results:Essential oil at 800 ppm showed larvicidal activity at 24 h with lethal values of LC_(50) and LC_(90) of 267.33 and 493.63 ppm.The hvdrolat at 20% and 10% on 2nd stage larvae showed 100%effectiveness after 24 h.The aqueous extract at 10% had a relative growth index of 0.58.while the ethanolic and methanolic extract obtained values of 0.76 and 0.70 and control reached 0.99.Larvae treated with 10% of methanol,ethanol and aqueous extract showed a reduction in larval duration of 5.00,2.20 and 4.35 days;ethanol extract at 1% provoke decrease of 2.40 days in the development and exhibited an increment of 3.30 days when treated with 0.01%.Aqueous,ethanol and methanol extracts at 10%reduced in 6.15,3.42 and 5.57 days pupal development.The main compounds were diallyl disulfide(50.05%),diallyl sulfide(11.77%) and disulfide di-2-propenyl(10.37%).Conclusions:The study demonstrated for the first time,the larvicidal activity of the essential oil and hydrolat of Pseudocalymma alliaceum:aqueous,ethanol and methanol extracts inhibited the normal growth and development of mosquito larvae,prolonging and delaying larval and pupal duration.

Reversion of malignancy in human gastric cancer MKN-45 cells through the transfection of transforming growth factor-β type Ⅱ receptor gene

Human gastric cancer MKN-45 cells which are resistant to TGF-β growth inhibition and possess TGF-β type Ⅰ and type Ⅲ receptors, but not type Ⅱ receptors, have been used as a model system to reconstitute these caflcer cells with TGF-β RII cDNA. The results of these experiments indicated that the reexpression of TGF-g RII gene in MKN-45 cells can restore their sensitivity to TGFβ growth inhibition, decrease their growth rate, reduce their cloning efficiency in soft agar and tumorigenicity in nude mice in stable transfectants, in comparison with their control MKN-45 cells. Among different RII transfectants,their difference in the changes of these parameters, as a result of the regain of autocrine negative growth control by TGF-β, is roughly proportional to their level of expression of transfected RII mRNA. From these data, it is concluded that the inactivation of TGF-β RII gene is related to the escape of growth control by TGF-β in MKN-45 cells. The importance of the study of the interplay of TGF-β and its receptor system in the negative growth control of gastric cancer, and possibly also of other cancers, is discussed.