ABSTRACT
Consideramos esta semblanza del Dr. Jorge Díaz Mayo, fallecido recientemente, un digno homenaje a un querido profesor de este Hospital "Enrique Cabrera"; admirado y respetado por todos, quien, por su dedicación profesional, constituyó un ejemplo no solo para los que trabajaron a su lado, sino para los que, de forma directa o indirecta, conocieron sus excelentes cualidades como médico y como ser humano. Su trayectoria como estudiante fue culminada como diplomado en Doctor en Medicina, en cuyo perfil se distinguiría con una prolífera labor durante años en varios centros médicos, en los cuales mantuvo siempre los principios y ética de médico íntegro y revolucionario, además de destacarse en actividades colaterales, políticas, deportivas y en la defensa al país. Su labor como dirigente fue ejemplar.
This is to pay tribute to the recently deceased professor Dr. Jorge Díaz Mayo, who was admired and respected by everyone during all his work at "Enrique Cabrera" Hospital. He was also an example of professional dedication not only to all the persons who worked directly with him but to those who, directly or indirectly, knew about his excellent qualities as a doctor and a human being. He ended up his student´s formation with the Degree of Doctor of Medicine. He distinguished himself for his prolific work in different medical centers for many years and he always kept the principles and ethics of upright and revolutionary doctor. He also distinguished himself for his collateral, political, and sports activities, as well as other actions for homeland defense. He was considered an inspiring leader.
ABSTRACT
Objective::To determine the chloroplast genomes of Liriope spicata var. prolifera, Ophiopogon japonicus in Sichuan and Zhejiang, analyze their sequence characteristics and complete the screening of specific DNA barcodes. Method::The chloroplast genomes of L. spicata var. prolifera, O. japonicus in Sichuan and Zhejiang were sequenced, spliced and annotated through high-throughtput sequencing technology, and the structural characteristics and phylogenetic relationships of chloroplast genomes were analyzed by bioinformatics. Result::The total length of chloroplast genome of L. spicata var. prolifera was 155 998 bp, the total content of guanine and cytosine (GC) was 37.7%, and 85 protein-coding genes, 37 transfer RNA (tRNA) genes and 8 ribosomal RNA (rRNA) genes were successfully annotated, a total of 274 simple sequence repeats (SSRs) were detected, the number of codons encoding leucine was the most, while the number of codons encoding tryptophan was the least. The total length of chloroplast genome of O. japonicus in Sichuan province was 156 078 bp, the total content of GC was 37.8%, and 85 protein-coding genes, 37 tRNA genes and 8 rRNA genes were successfully annotated, a total of 265 SSRs were detected, the number of codons encoding leucine was the most, while the number of codons encoding tryptophan was the least. The total length of chloroplast genome of O. japonicus in Zhejiang province was 156 207 bp, the total content of GC was 37.7%, and 85 protein-coding genes, 37 tRNA genes and 8 rRNA genes were successfully annotated, a total of 274 SSRs were detected with the highest number of codons encoding leucine and the lowest number of codons encoding tryptophan. Conclusion::The phylogenetic trees show that compared with O. japonicus in Sichuan province, L. spicata var. prolifera is more closely related to O. japonicus in Zhejiang province. The variation of non-coding regions of L. spicata var. prolifera, O. japonicus in Zhejiang and Sichuan is greater than that in the coding region. The entire chloroplast genome can be used as a super barcode for identifying species of Ophiopogon and Liriope.
ABSTRACT
OBJECTIVE：To study the effects of loganin on the prolife ration and apoptosis of liver cancer HepG 2 cells，and to explore its mechanism. METHODS ：CCK-8 assay was used to detect the effects of different concentrations （10，25，50，100， 150，200，300，400 µg/mL）of loganin on the proliferation activity of HepG 2 cells for 24 and 48 h. HepG 2 cells were divided into control group ，loganin low-concentration ，medium-concentration and high-concentration groups （50，100，150 μ g/mL）. After treated for 24 h，morphological changes of apoptosis of cells were detected by Hoechst 33342 fluorescence staining. The apoptosis and cycle distribution of cells were detected by flow cytometry. Western blotting was used to detect protein expression of Cyclin D1， PCNA， Bcl-2， Caspase-3， Cleaved-Caspase-3， Caspase-9 and Cleaved-Caspase- 9. RESULTS ： Loganin inhibited the proliferation of HepG 2 cells，in concentration-dependent trend. Compared with control group ，apoptosis as pyknosis and fragmentation occurred ，and the apoptosis rate increased significantly in loganin low-concentration ，medium-concentration and high-concentration groups （P＜0.01）；the cell were mainly blocked in S phase ；relative protein expression of Cyclin D 1，PCNA and Caspase- 3 were significantly decreased ，while that of Cleaved-Caspase- 3 were significantly increased in loganin low- concentration， medium-concentration and high-concentration groups （P＜0.05 or P＜0.01）； relative protein expression of Cleaved-Caspase-9 were increased significantly ，while that of Bcl- 2 and Caspase- 9 were decreased significantly in loganin medium-concentration and high-concentration groups （P＜0.05 or P＜0.01）. CONCLUSIONS ：Loganin can significantly inhibit the proliferation and induce apoptosis of HepG 2 cells，the mechanism of which may be associated with inhibiting Bcl- 2 protein expression and promoting Caspase- 3，Caspase-9 activation.
ABSTRACT
Aim To investigate the effects of Periplane-ta americana extract Ento-A on the immune function in immunosuppressed mice . Methods Immunosup-pressed mouse model was induced by intraperitoneal injection of cyclophosphamide in KM mice .To evalu-ate the effects of Ento-A on the immune function in im-munosuppressed mice , neutral red method and MTT assay were used respectively to detect the effects of En-to-A on the phagocytosis of peritoneal macrophages and T cell proliferation rate in mice; with sheep red blood cell as immunogen , the effects of Ento-A on the pro-duction of serum hemolysin were evaluated;peripheral blood was tested and immune organ index calculated . Results Compared with model control group , the high, medium and low doses of Ento-A could improve the expression of serum hemolysin in immunosup-pressed mice ( P<0.01 ) , and increase the spleen in-dex(P<0.01) and thymus index (P>0.05), signifi-cantly increased the content of WBC ( P<0.01 ) , PLT ( P<0.01 ) , HGB ( P<0.01 ) , while the contents of RBC was on the rise , with no significant difference ( P>0.05 ) in peripheral blood , significantly enhanced phagocytic function and T lymphocyte proliferative abil-ity in a dose-dependent manner ( P<0.01 ) .Conclu-sion Ento-A can enhance the immune function of im-munosuppressed mice .
ABSTRACT
Seven flavonoid compounds, kaempferol (1), quercetin (2), quercetin-3-O-β-D-glucopyranoside (3), kaempferol-3-O-β-D-glucopyranoside (4), kaempferol-3-O-α-L-rhamnoside (5), quercetin-3-O-sophoroside (6) and quercetin-3-O-rutinoside (7), were isolated from the methanolic extract of leaves of Kalanchoe prolifera. Compounds 1-7 were isolated for first time from this plant. These compounds were evaluated their cytotoxic activity against P-388 murine leukimia cells in vitro. Among those compounds kaempferol (1) and quercetin (2) showed strongest cytotoxic activity with IC₅₀ values of 4.45 ± 0.05 and 6.28 ± 0.02 µg/mL, respectively.
Subject(s)
Crassulaceae , Flavonoids , In Vitro Techniques , Kalanchoe , Methanol , Plants , QuercetinABSTRACT
AIM:To identify the expression of fermitin family homolog 2 (FERMT2) in hepatocellular carci-noma ( HCC) tissues and the effect of FERMT 2 on the cell growth and related protein expression .METHODS:Real-time PCR and immunohistochemistry were used to detect FERMT 2 expression in the HCC tissues .The technique of CRISPR/Cas9 was applied to construct stable FERMT2 knockout MHCC97H cell line.WST-1 assay and flow cytometry were used to measure the cell viability , cell-cycle distribution and cell apoptosis .Western blot was used to determine the expression of related proteins in the MHCC97H cells.RESULTS:In HCC tissues, the expression level of FERMT2 was higher than that in adjacent liver tissues (P<0.05).High expression of FERMT2 was significantly correlated with postoperative recurrence of tumor.Knockout of FERMT2 gene evidently inhibited MHCC97H cell viability and accelerated cell apoptosis .Mean-while, the expression levels of proliferating cell nuclear antigen , cyclins, cyclin-dependent kinases 2 and anti-apoptotic fac-tors were significantly downregulated in MHCC97H cells with FERMT2 knockout (P<0.05).CONCLUSION:FERMT2 may function as a promoter of hepatocarcinogenesis and progression via regulating the cell viability , cell-cycle distribution and cell apoptosis , which is related with the expression of cell cycle regulators and anti-apoptotic factors .
ABSTRACT
Aim To investigate the role of TGF-β3 in the anti-proliferation effect of ursolic acid(UA)in co-lon cancer cells and the possible molecular mechanism underlying this effect.Methods We introduced crys-tal violet staining,flow cytometry and Western blot as-say to determine the effect of UA on proliferation and apoptosis in HCT1 1 6 cells.The levels of TGF-β3, Smad2 /3 and β-catenin in HCT1 1 6 cell were evaluated by RT-PCR and Western blot.Finally,TGF-β3 inhibi-tor and recombinant adenovirus,and luciferase reporter assay were used to analyze the possible mechanism through which TGF-β3 mediated the anti-cancer effect of UA in HCT1 1 6 cells.Results UA inhibited the proliferation and induced apoptosis apparently in HCT1 1 6 cells.UA down-regulated TGF-β3 both in mRNA and in protein level.Meanwhile,UA decreased the phosphorylation of Smad2 /3 concentration depend-ently,although no significant effect was found on the total protein level of Smad2 /3 in HCT1 1 6 cells.Over-expression of TGF-β3 attenuated the inhibitory effect of UA on the proliferation of HCT1 1 6 cells,while the TGF-β3 inhibitor potentiated this effect. UA sup-pressed the transconduction of Wnt/β-catenin signaling in HCT1 1 6 cells through decreasing the level of β-catenin.Exogenous expression of TGF-β3 increased the level of β-catenin and partly reversed the UA-in-duced decrease of β-catenin.However,TGF-β3 inhib-itor potentiated the inhibitory effect of UA on β-catenin in HCT1 1 6 cells.Conclusion The anti-proliferation activity of UA in colon cancer may be partly mediated through down-regulating TGF-β3 to suppress Wnt/β-catenin signaling at least.
ABSTRACT
Aim To investigate the promoting apoptosis effect of artesunate( ART) on human colon cancer Lovo cells and its mechanisms. Methods MTT assay was performed to determine the anti-proliferative effect of artesunate. Flow cytometry assay and electron micros-copy( EM) were used to evaluate the apoptotic effect of artesunate. Luciferase reporter assay was introduced to measure the activation of Wnt/β-catenin pathway. Western blot was used to detect the pathway-related protein levels of β-catenin, GSK-3β,c-Myc and apop-tosis-related protein level of casepase-3 . Results Compared with the control group, the inhibitory rate of cell proliferation at 72 h and 320 μmol·L-1 ART was (78. 99 ± 1. 95 )% ( F =898. 301, P =0. 000 ); the cell apoptotic rate at 24 h and 160 μmol · L-1 ART was(19. 00 ± 0. 05)% and morphological signs of cell apoptosis were found by EM;the transcriptional activi-ty of TCF4/LEF at 24 h and 160 μmol·L-1 ART was (0. 36 ± 0. 30)%(F =470. 954,P <0. 01); the ex-pressions of caspase-3 and GSK-3β were significantly increased, whileβ-catenin and c-Myc were significant-ly decreased when treated with different concentrations of ART for 48 h ( P <0. 01 ) . Conclusion ART may significantly inhibit proliferation and promote apoptosis of Lovo cells probably by inactivating Wnt/β-catenin pathway.
ABSTRACT
Objective To observe the neural protection of 3-n-butylphthalide (NBP) injection in focal cerebral ischemia-reperfusion rats. Methods 160 male Sprague-Dawley rats were randomly divided into sham group (n=10), ischemia-reperfusion group (IR group, n=50), high-dose NBP treatment group (high-dose group, n=50), middle-dose NBP treatment group (middle-dose group, n=25) and low-dose NBP treatment group (low-dose group, n=25). The later 4 groups were occluded the middle cerebral artery for 2 hours and reperfused. The sham group was sacrificed 24 hours after operation, and the other groups at 6, 12, 24, 48 and 72 hours after reperfusion, in which 5 of them were stained with TdT mediated dUTP Nick End Labeling (TUNEL) to observe the neuronal apoptosis, and immunohistochemistry to observe the expression of silent information regulation 2 homolog 1 (SIRT1) and peroxisome proliferator-activated receptorγcoactivator-1α(PGC-1α);the other 5 of sham group, IR group and high-dose group were observed with quantitative real-time PCR of SIRT1 and PGC-1α. Results Compared with the IR group, the number of apoptotic cells decreased and the SIRT1 and PGC-1αpositive cells increased in all NBP groups at each time (F>160.60, P4.13, P<0.01). Conclusion NBP can protect brain from apoptosis in focal cerebral ischemia-reperfusion rats, which may relate to more ex-pression of SIRT1 and PGC-1α.
ABSTRACT
AIM: To investigate the effect of inhibiting myosin light chain kinase ( MLCK) on endothelin-1 (ET-1) induced proliferation and apoptosis of rat pulmonary artery smooth muscle cells (PASMCs).METHODS: Rat PASMCs were cultured and stimulated with ET-1.The cells were randomly divided into control group , ET-1 group and ET-1+MLCK inhibitor group (ET-1+M).Western blotting, MTT assay, [3H]-TdR incorporation and flow cytometry were employed to test the expression of myosin light chain (MLC) and MLCK, cell proliferation, cell cycle and apoptotic rate of PASMCs ,respectively .The phosphorylation of MLC was determined by glycerol-PAGE coupled with Western blotting .RE-SULTS:Compared with control group , the protein expression of MLCK and MLC phosphorylation significantly enhanced af -ter ET-1 stimulation.ET-1 markedly induced the proliferation and decreased the percentage of apoptotic rate in the PASMCs.However, pretreatment with ML-7, a MLCK inhibitor, significantly reversed the above effects induced by ET-1. CONCLUSION:MLCK inhibitor effectively inhibits the ET-1-induced proliferation and the cell cycle progression .
ABSTRACT
Objective To investigate the relationship between CYP2J2*7 mutation(G-76T) and coronary heart disease (CHD) in Chinese Hanpopulation and to study the effects of CYP2J2 geneover-expressionon the proliferation and migrationof aortic smooth muscle cells of ApoE-/- mice. Methods CYP2J2*7 genotype was detectedin 500 patients with CHD and 478 controlsubjects by the Polymerase Chain Reaction-Restriction Frag-ment Length Polymorphism (PCR-RFLP). Culturedaortic smooth muscle cells of ApoE-/- mice were divided into control group, sham transfectiongroup and CYP2J2 over-expression group. Cell proliferation and migration were investigated after CYP2J2 over-expressionby MTS and Transwell assay. Results The frequency of CYP2J2*7 in CHD group was significantly higher than that incontrol group (10.00% vs. 6.49%, P = 0.046). Same is the case in female cases(P = 0.026). Compared with these of aortic smooth muscle cells incontrol group and sham trans-fectiongroup, the cell proliferation in 24, 48, 72 h, and the cell migration in 48 h after CYP2J2 over-expression in CYP2J2 group were significantly suppressed. Conclusions CYP2J2*7 mutation might increase the risk of CHD in Chinese Han population. CYP2J2 over-expression can suppress the proliferation and migration of aortic smooth muscle cells and CYP2J2 might have the effect of anti-atherosclerosis.
ABSTRACT
Aim To investigate the anti-proliferating effect of tetrandrine ( Tet ) on colon cancer cells and its possible molecular mechanism. Methods We intro-duced crystal violet staining and flow cytometry to ana-lyze the effect of Tet on proliferation in LoVo cells. Flow cytometry was used to detect the effect of Tet on apoptosis in LoVo cells. Western blot assay was taken to analyze the effect of Tet on the expression of insulin-like growth factor binding protein 5 ( IGFBP5 ) . Final-ly, luciferase reporter assay, recombinant adenovirus mediated over-expression or silence of IGFBP5 were used to analyze the possible role of IGFBP5 in the anti-proliferating effect of Tet on colon cancer cells. Re-sults Crystal violet staining and flow cytometery anal-ysis results showed that Tet could exert an anti-prolifer-ating effect and induce apoptosis in LoVo cells. Tet de-creased the expression of IGFBP5 in a concentration-dependent manner. Tet inhibited the transcriptional ac-tivity of pTOP-luc reporter, which could be reversed by exogenous expression of IGFBP5 mostly. Similar results were found in the expression of c-Myc, but IGFPB5 knockdown couldn’ t reverse this effect. Conclusion Tet can inhibit the proliferation of colon cancer cells, and this effect may be mediated by down-regulating the expression of IGFBP5 to inhibit Wnt/β-catenin signa-ling transduction partly.
ABSTRACT
Pulmonary artery hypertension (PAH ) is a chronic progressive disease characterized by a persistent elevation of pul-monary vascular pressure,and the disease would limit the right ventricular function severely,fail the organ and even lead to death in the end.The histopathological change of PAH is fea-tured by the restructuring of pulmonary vessels,and the abnor-mal reproduction of pulmonary artery smooth muscle cells (PASMCs)in peripheral vessels is the major pathological basis of pulmonary vascular restructuring.This paper mainly reviews the research advances on signal transduction mechanisms and their inhibitors in promoting the proliferation of pulmonary artery smooth muscle cells.
ABSTRACT
Abstrac:Aim To study the effect of hydroxysafflor yellow A ( HYSA ) on the proliferation of vascular smooth muscle cells ( VSMCs) and the related molecu-lar mechanism. Methods The inhibitory effects of hydroxysafflor yellow A on VSMC proliferation was de-tected using cell culture, MTT assay, Western blot and immunohistochemical staining. Results The results showed that HYSA inhibited cell proliferation induced by PDGF in a dose-dependent (5,10,20,40 μmol· L-1 ) manner, reduced proliferating cell nuclear anti-gen ( PCNA ) expression and blocked PDGFR-MEK-ERK1/2 signaling pathway activated by PDGF in VSMCs. Conclusion HYSA inhibits VSMCs prolifer-ation via reducing the expression of PCNA and blocking signal transduction of MEK-ERK1/2 in VSMCs.
ABSTRACT
Aim To investigate the anti-proliferation effect of resveratrol (Res)on human colon cancer cells and dissect the possible mechanism underlaying this effect.Methods We introduced crystal violet staining and Western blot to analyse the anti-proliferation effect of Res on HCT1 1 6 cells.Then,we used flow cytome-try and Western blot assay to detect the Res induced apoptosis in HCT1 1 6 cells.Next,we employed the well established TCF4 /LEF luciferase reporter to meas-ure the effect of Res on Wnt/β-catenin signaling trans-duction.Finally,we took Western blot and PCR assay to analyse the effect of Res on the expression of β-cate-nin in HCT1 1 6 cells.Results Crystal violet staining and Western blot analysis showed that Res could inhib-it the proliferation of HCT1 1 6 cells in a concentration-and time dependent fashion.What’s more,Res could promote apoptosis in HCT1 1 6 cells.The transcriptional activities of TCF4 /LEF reporter were reduced by Res in a concentration-dependent fashion (P <0.05 when the concentration of Res was 20 μmol·L -1 ,and P <0.01 when the concentration of Res was 40 μmol·L -1 or 80 μmol·L -1 ).Res could decrease not only the protein level of β-catenin in HCT1 1 6 cells,but also the mRNA expression of β-catenin.Conclusion Res can inhibit the proliferation of HCT1 1 6 cells,which may be mediated at least by down-regulating the ex-pression of β-catenin to inhibit the Wnt/β-catenin sig-naling transduction.
ABSTRACT
Aim To investigate the protective effect of cinnamic aldehyde ( CA ) on hormone-induced osteo-clasts proliferation and bone resorption in vitro and its molecular mechanisms. Methods RAW264. 7 cells induced into osteoclast were treated with RANKL and M-CSF and then were divided into control group, dexa-methasone ( DEX ) group and different doses of CA (11. 6, 23. 2, 46. 4 μg·L-1 ) groups. OCs were ob-served after tartrate resistant acid phosphatase( TRAP) staining. The cell proliferation was determined by MTT assay at different time points. The expression levels of TRACP5 b in cell cultured supernatants were measured by ELISA. RT-PCR technique was applied to examine the transcriptional levels of RANK and NFATc1 . Re-sults In MTT assay, the proliferation of osteoclasts stimulated by dexamethasone was promoted seriously compared with negative control group ( P < 0. 05 ) . Meanwhile, DEX could strengthen the content of TRACP5 b and up-regulate the expressions of RANK and NFATc1 mRNA. After administration of CA, the proliferation was inhibited, while the enhanced expres-sion of TRAP5b was reversed,and the over-expressions of RANK and NFATc1 mRNA were obviously down-regulated in a time-and-dose-dependent manner ( P <0. 05 ) . Conclusion The results suggest that CA in-hibits proliferation and bone resorption of osteoclast in-duced by DEX, which may be mediated by down-regu-lation of RANK and NFATc1 mRNA.
ABSTRACT
Aim To investigate the effects of isoliquiri-tigenin(ISL)on C6 glioma cell proliferation and differ-entiation.Methods C6 glioma cells’viability and proliferation were respectively measured by SRB test. Colony formation of C6 glioma cells from different groups was assayed.After culturing the cells from each group,giemsa staining was used to observe cell mor-phology.RT-PCR was applied to detect mRNA expres-sion of GFAP.Western blot was applied to detect the expression of GFAP.Results ISL effectively inhibited the viability of C6 glioma cells when compared with the control group in a concentration-dependent manner (P<0.01).The morphological observation under light mi-croscope showed that:in the control group,most of the undifferentiated C6 cells showed long fusiform and po-lygonal shape.Compared to the control group,the C6 cells treated with ISL revealed alteration in morphology such as astrocytes with smaller smooth,round body and much finer longer,tapering processes.The cloning for-mation rate detection revealed that:the colonies in the control group semerged earlier and were larger than those experimental ones,the cloning formation rate was higher,while almost no effective cells colony emerged in ISL treated groups(P <0.01 ).Western blot and RT-PCR analysis showed that GFAP expression in the ex-perimental groups increased(P <0.01).Conclusion ISL may inhibit the proliferation of C6 glioma cells and induce their differentiation.
ABSTRACT
Objective To investigate the effects and the possible mechanism of insulin like growth factor-1 ( IGF1 ) on proliferation and migration of human umbilical vein endothelial cells ( HUVEC) in high glucose. Methods Cultured HUVEC were divided into 5 groups( control group, high glucose group, hypertonic group, high glucose+IGF 1 group, high glucose+IGF1+ AZD5363 group) . The proliferation was detected by methyl thiazolyl tetrazolium ( MTT) , the migration was detected by transwellchamber. The mRNA expressions of serine/threonine kinase( Akt) and FoxO1 were detected by real-time PCR. The protein expressions of FoxO1/p-FoxO1 and Akt/p-Akt were detec-ted by immunohistochemisty. Results Compared with the control group, the rate of cell survival in high glucose group decreased significantly(P<0. 05), the transitional was decreased significantly(P<0. 05), the mRNA ex-pression of FoxO1 was increased significantly( P<0. 05 ) , the protein expressions of FoxO1/p-FoxO1 and Akt/p-Akt were increased. Compared with high glucose group, the cell survival rate and migration rate in high glucose+IGF1 group were increased significantly, the mRNA expression of FoxO1 was decreased significantly(P<0. 05), the protein expressions of FoxO1/p-FoxO1 and Akt/p-Akt were decreased. Compared with high glucose group, the cell survival rate and migration rate in high glucose+IGF1+AZD5363 group had no significant difference, the mR-NA expression of FoxO1 was decreased significantly ( P<0. 05 ) , the protein expressions of FoxO1/p-FoxO1 had no significant difference. The mRNA expression of Akt in five groups had no significant difference. Conclusion IGF1 can improve the proliferation and migration of endothelial cell in high glucose. It shows that IGF1 acts through Akt to promote FoxO1 phosphorylation.
ABSTRACT
Objective To investigate the effects of substrate stiffness on the proliferation of human umbilical vein endothelial cells ( HUVEC) during dengue virus infection.Methods Polyacrylamide gels were prepared for cell culture [(0±4) kPa].The proliferation of HUVEC cultured on substrates with differ-ent stiffness was determined by using 3-(4,5-diethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfo-phenyl)-2H-etrazolium,inner salt ( MTS) assay.The cycle and apoptosis of HUVEC were determined by flow cytometry analysis.Dengue virus serotype 2 (DENV-2) strains were propagated and identified by con-ventional assays.The HUVEC were infected with DENV-2 strains at a MOI of 10 and cultured on traditional plastic and hydrogel substrates, respectively.The levels of nitric oxide (NO) and endothelin-1 (ET-1) were detected by nitric acid reductase assay and double antibody sandwich ELISA.Results Young′s modulus E value of the hydrogels was (3030 ±0.44) Pa.The proliferation of HUVEC and the expression of NO and ET-1 were enhanced along the increased substrate stiffness.However, no significant differences with the cell cycle and apoptosis were observed between cells cultured on different substrates.Conclusion The stiffness of substrates affected not only the proliferation of HUVEC, but also the release of cytokines during DENV-2 infection.The development of dengue fever was associated with the decreased secretion of vascular active substances as a result of blood vessel injury.The establishment of hydrogel substrates as the model of vascu-lar basement membranes might provide a new way for the in vitro investigation of the pathogenesis of DENV infection.
ABSTRACT
Aim To investigate the effects of Guaveno-ic acid (GA)on proliferation,insulin synthesis and secretion in INS-1 cells and their possible mechanism. Methods INS-1 βcells were cultured in vitro.Control group,medium group,model group,drug groups and positive group were set.INS-1 cells were treated with GA (0.3,1 ,3,1 0,30 nmol·L -1 )for 48 h.The cell proliferation was tested by MTT assay.Acid-alco-hol was used to extract the insulin in the cells and the amount of insulin synthesis of INS-1 cells was tested by RIA.5.6,1 6.7 mmol·L -1 glucose was used to chal-lenge INS-1 cells for 1 h to the insulin secretion model (BIS and GSIS)was tested,and the insulin secretion of INS-1 cells was tested via RIA.The mRNA expres-sion of insulin gene,PDX-1 and MafA was tested by q-PCR.Results Compared with medium group,GA could promote the proliferation of INS-1 cells signifi-cantly (P <0.01 )and promote the synthesis of insulin in INS-1 cells significantly (P <0.01 ).GA(0.3 ~30 nmol· L -1 )could promote the BIS,GSIS of INS-1 cells significantly (P <0.05 or P <0.01 ).GA (3,30 nmol·L -1 )could up-regulate the mRNA expression of insulin gene,PDX-1 ,MafA in INS-1 cells signifi-cantly (P <0.01 ).Conclusions GA could signifi-cantly improve the proliferation of INS-1 cells and pro-mote the insulin synthesis and secretion of INS-1 cells, which may be associated with up-regulation of insulin gene,PDX-1 ,MafA mRNA expression.