An increasing number of monopartite begomoviruses are being identified that a satellite molecule (DNAbeta) is required to induce typical symptoms in host plants. DNAbeta encodes a single gene (termed betaC1) encoded in the complementary-sense. We have produced transgenic Nicotiana benthamiana and N. tabacum plants expressing the betaC1 gene of a DNAbeta associated with Tomato yellow leaf curl China virus (TYLCCNV), under the control of the Cauliflower mosaic virus 35S promoter. Transgenic plants expressing betaC1 showed severe developmental abnormalities in both species. Microscopic analysis of sections of both transgenic and non-transgenic N. tabacum leaves showed abnormal outgrowths of transgenic N. tabacum to be due to disorganized cell division (hyperplasia) of spongy and palisade parenchyma. Immuno-gold labeling of sections with a polyclonal antibody against the betaC1 protein showed that the betaC1 protein accumulated in the nuclei of cells. The possible biological function of the betaC1 protein was discussed.
Cell Division ; physiology ; Cell Nucleus ; genetics ; metabolism ; ultrastructure ; Cells, Cultured ; DNA, Viral ; genetics ; Geminiviridae ; genetics ; Plant Diseases ; genetics ; virology ; Plant Leaves ; cytology ; genetics ; growth & development ; metabolism ; Plants, Genetically Modified ; growth & development ; metabolism ; Recombinant Proteins ; metabolism ; Tobacco ; cytology ; growth & development ; metabolism ; ultrastructure ; Viral Proteins ; genetics ; metabolism

Objective To study their chemical components of xianglingcao(Crotalaria ferruginea) and identify their chemical structures.Methods The compounds were isolated by chromatography and their structures were identified by spectral analysis and compared with the published data.Results Ten compounds were isolated and identified as vomifolilol(Ⅰ),genistein(Ⅱ),p-hydroxybenzoic acid(Ⅲ),5,7-dihydroxy-4-methoxy-flavone-7-O-?-D-glycopyranoside(Ⅳ),octadecoic acid(Ⅴ),octacosanol(Ⅵ),?-stitosterol(Ⅶ),?-daucosterol(Ⅷ),stigmasterol(Ⅸ),and ?5,22 stigmasterol-3-O-?-D-glycopyranoside(Ⅹ).Conclusion The ten compounds are obtained from xianglingcao for the first time.

Breast Neoplasms ; metabolism ; pathology ; surgery ; Carcinoma ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Female ; Fibroma ; metabolism ; pathology ; surgery ; Humans ; Keratins ; metabolism ; Middle Aged ; Vimentin ; metabolism

Objective To study the effects of toys for laboratory animals on reproductive performance and growing development of experimental mice.Methods ICR mice were fed with toys, the informations of reproductive performance and growing development were recorded.Results All the data of reproductive performance of the test group were higher than the control group except the number of newborn mice, and showed significant difference ( P <0.05 ) .The data of growing development of the test group were higher than the control group too, but showed no significant difference ( P >0.05).Conclusion Toys for laboratory animals have good effects on reproductive performance and growing development of mice, and suggested to be used into the process of experimental mice raising.

OBJECTIVETo study the effect of Tongluo Xingnao effervescent tablets on learning and memory capacity and expression of Na(+)-K(+)-ATPase in hippocampus of rats with chronic cerebral ischemia-induced learning and memory dysfunction model.

METHODThe 2-VO method was used to establish sd rat model learning and memory dysfunction induced by chronic cerebral ischemia. The 50 rats in the successfully established model were randomly divided into the model control group, the Dihydroergotoxine Mesylate tablets group (0.7 mg x kg(-1), Tongluo Xingnao effervescent tablets high dose (7.56 g x kg(-1)), middle dose (3.78 g x kg(-1)) and low dose (1.59 g x kg(-1)) groups and the sham operation group (n = 10) as the control group. The groups were orally given 10 ml x kg(-1) x d(-1) drugs for consecutively 90 days. On the 86th day, Morris water maze was adopted for them. On the 90th day, a leaning and memory capacity test was held. The brain tissues were fixed with 10% formaldehyde and observed for pathomorphism after routine slide preparation and staining. The expression of hippocampal Na(+)-K(+)-ATPase was detected with immunohistochemistry and image quantitative analysis.

RESULTCompared with the model group, all of Tongluo Xingnao effervescent tablets groups showed significant decrease in the escape latency at the 5th day in the Morris water maze, and notable increase in the frequency of the first quadrant dwell, the frequency passing the escape platform and the frequency entering effective area (p < 0.05). According to the pathomorphological detection, the control group showed a significantly higher pathological score than the sham operation group (p < 0.01), the middle dose group showed a significantly lower pathological score than the model group (p < 0.05). According to the immunohistochemistical detection, the model control group showed a remarkably lower mean OD value of Na(+)-K(+)-ATPase than the sham operation group (p < 0.05), high and middle dose groups showed a significantly higher mean od value than the model control group (p < 0.01).

CONCLUSIONTongluo Xingnao effervescent tablets can improve the learning and memory capacity, reduce pathological changes of hippocampal tissues of rats with chronic cerebral ischemia-induced learning and memory dysfunction model, and promote the expression of Na(+)-K(+)-ATPase in hippocampus.

Animals ; Brain Ischemia ; drug therapy ; enzymology ; genetics ; psychology ; Chronic Disease ; drug therapy ; psychology ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hippocampus ; drug effects ; enzymology ; Humans ; Learning ; drug effects ; Male ; Memory ; drug effects ; Rats ; Rats, Sprague-Dawley ; Sodium-Potassium-Exchanging ATPase ; genetics ; metabolism ; Tablets ; administration & dosage

AIM:To predict and identify an HLA-A3 supertype-restricted cytotoxic T-lymphocyte(CTL) epitope derived from MAGEC2,which is utility in epitope design for the development of HLA-based vaccines and immuno-therapeutics.METHODS:HLA-A3 epitopes from MAGEC2 protein were predicted by BIMAS, SYFPEITHI and IEDB. The binding affinity of the peptides to HLA-A*03 molecule was evaluated by T2A3 cell binding assay.ELISPOT assay was used to investigate the ability of the peptides inducing specific restricted CTLs to release interferon -γ(IFN-γ).The ability of the peptides to induce T-cell response was investigated by cytotoxicity assay in vitro.RESULTS:The candidate peptides P147,P167, P196, P229 and P251 showed moderate affinity toward HLA-A3 molecule.ELISPOT assay showed that P167,P196 and P251 were able to induce specific CTLs and higher levels of IFN-γwere released.The CTLs induced by P196 and P251 were able to lyse target cells.CONCLUSION:The peptides P196 and P251 have higher binding affinity with HLA-A3 and retain immunogenicity.They are excellent HLA-A3-restricted CTL epitopes from tumor antigen MA-GEC2,which could serve as new candidates towards antitumor peptide vaccines.

AIM: To investigate the different effects of salvianolic acid and notoginseng triterpenes on proliferation, angiogenesis and expression of vascular endothelial growth factor in EA-hy926 cells in vitro. METHODS: EA-hy926 cells were cultured in vitro. Salvianolic acid and notoginseng triterpenes at concentrations of 0.4, 0.8 and 1.2 mg·L(-1) were used to culture EA-hy926 cells. EA-hy926 cells in a blank control group were grown in culture solution only. Viability of cells was assessed by CCK-8, and after treated for 12 h, capillary-like structures were examined. After 24 h culture, the expression of VEGF was detected by real-time PCR. RESULTS: Salvianolic acid at 0.4, 0.8 mg·L(-1), the same as notoginseng triterpenes, increased VEGF content in EA-hy926 cells. Expression of VEGF protein in the salvianolic acid at 1.2 mg·L(-1) group, was up-regulated as compared with notoginseng triterpenes group (P < 0.05). CONCLUSION Salvianolic acid and notoginseng triterpenes can promote EA-hy926 cell proliferation, angiogenesis and expression of VEGF protein. This analysis also provided evidence that salvianolic acid had the better effects as compared with notoginseng triterpenes.
Alkenes ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Coronary Stenosis ; drug therapy ; genetics ; metabolism ; physiopathology ; Drugs, Chinese Herbal ; pharmacology ; Endothelial Cells ; drug effects ; metabolism ; Humans ; Neovascularization, Pathologic ; drug therapy ; genetics ; metabolism ; physiopathology ; Panax notoginseng ; chemistry ; Polyphenols ; pharmacology ; Triterpenes ; pharmacology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

AIM: To investigate and analyze the changes of corneal biomechanics of normal eyes,forme frusta keratoconus eyes,subclinical keratoconus eyes and clinical keratoconus eyes by Corneal visualization Scheimpflug technology (Corvis ST),and provide clinical basis for early diagnosis of keratoconus.METHODS: Case-control study.We randomly selected 40 normal eyes as normal group,15 forme frusta keratoconus eyes as forme frusta keratoconus group,23 subclinical keratoconus eyes as subclinical keratoconus group,and 40 clinical keratoconus eyes as keratoconus group.The biomechanical parameters of each group were measured by Corvis ST.The receiver operating characteristic(ROC) curves was plotted to distinguish keratoconus from the normal cornea.RESULTS: There was no significant difference in the parameters of biomechanics between normal group and forme frusta keratoconus group (P>0.05).Compared to normal group and subclinical keratoconus group,the parameters second applanation length(AL2),first velocity of applanation (AV1),central curvature radius at highest concavity (HC-radius),deformation amplitude (DA) were revealed statistically significant differences(P<0.05).The biomechanical parameters of the keratoconic group were significantly different from those of normal group except for the second velocity of applanation (AV2),time from the start until the highest concavity(HC-time),peak distance (PD).ROC curve showed that the DA(area under the curve:0.891±0.028) was the best predictive parameter to distinguish keratoconus from the normal eyes.CONCLUSION: The corneal biomechanical parameters of forme frusta keratoconus group are not changed compared with normal group.The changes between normal group and subclinical keratoconus group should combine with other technology to further improve subclinical keratoconic screening.Compared with normal corneas,keratoconus has a great change in biomechanics,which DA diagnosis of the highest efficiency.

ObjectiveTo compare the effect of three serological methods for detection of Yersina pestis F1 antibody.MethodsF1 antibody of Yersinapestis was detected with the methods of enzyme linked immunosorbent assay(EL1SA),indirect hemagglutination assay(IHA) and gold-immunochromatography assay (GICA),respectively.ResultsThe highest antibody titer was 1 ∶ 5120 by ELISA and 1 ∶ 640 by IHA.Meanwhile,the highest antibody titer of GICA was 1∶ 1280.ConclusionsEL1SA is the most sensitive method in detection of Yersina pestis F1 antibody.The sensitivity of GICA is low and that of IHA is the lowest of three serological methods.

OBJECTIVETo evaluate the expression of nitric oxide synthase III (NOS III) mRNA in the heart, aorta, kidney and liver of spontaneously hypertensive rats (SHR).

METHODSTwo hundred and ninety-four total RNA samples were obtained from the tissues of ventricle, aortic smooth muscle, kidney and liver of SHR and normotensive rats (Wistar-Kyoto rats, WKY). RNA array was used to determine the mRNA levels of NOS III of the two groups.

RESULTSCompared with WKY, the systolic blood pressure increased significantly in SHR at 6-week-old, 8-week-old, 10-week-old and 12-week-old [(158.50 +/-7.69 vs 108.67 +/-5.89) mmHg, (174.33 +/-4.46 vs 128.50 +/-4.97) mmHg, (198.00 +/-13.45 vs 142.00 +/-3.58) mmHg, (216.67 +/-8.91 vs 141.17 +/-4.92) mmHg, P<0.01], and the ventricle/body weight ratio was significant higher at 10-week-old and 12-week-old [(4.08 +/-0.17 vs 3.59 +/-0.11, 4.05 +/-0.18 vs 3.40 +/-0.19)mg/g, P<0.01]. In the heart tissue and the kidney, the mRNA levels of NOS III were significantly increased at 6-week-old, 8-week-old, 10-week-old and 12-week-old (1.12 +/-0.18 vs 0.90 +/- 0.15, 1.46 +/- 0.34 vs 1.06 +/-0.18, 1.66 +/- 0.31 vs 1.21 +/- 0.30, 1.98 +/- 0.40 vs 1.31 +/-0.38, P <0.05) and at 4-week-old, 6-week-old, 8-week-old, 10-week-old and 12-week-old (1.10 +/- 0.21 vs 0.81 +/-0.11, 1.28 +/-0.18 vs 0.95 +/-0.13,1.31 +/-0.23 vs 0.99 +/-0.23, 1.70 +/-0.30 vs 1.08 +/-0.25, 1.83 +/-0.33 vs 1.15 +/-0.20, P<0.05 or P<0.01), respectively. There was no significant difference of the NOS III expression in the liver and no significant signals were detected in the aortic smooth muscle.

CONCLUSIONThe results provide the evidence of the increased expression of NOS III in different tissues in SHR and suggests the progressive nature of essential hypertension.

Animals ; Hypertension ; enzymology ; genetics ; Kidney ; enzymology ; Liver ; enzymology ; Male ; Myocardium ; enzymology ; Nitric Oxide Synthase ; biosynthesis ; genetics ; Nitric Oxide Synthase Type III ; Oligonucleotide Array Sequence Analysis ; methods ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY