MoFLP1, encoding a novel fungal fasciclin-like protein, is involved in conidiation and pathogenicity in Magnaporthe oryzae

Fasciclin family proteins have been identified as cell adhesion molecules in various organisms. In this study, a novel Magnaporthe oryzae fasciclin-like protein encoding gene, named MoFLP1, was isolated from a subtractive suppressive cDNA library and functionally analyzed. Sequence analysis showed that the MoFLP1 gene contains an open reading frame (ORF) of 1050 nucleotides encoding 349 amino acids with a calculated molecular weight of 35.85 kDa and a pI of 7.76. The deduced MoFLP1 protein contains a 17-amino acid secretion signal sequence and an 18-amino acid sequence with the characteristics of a glycosylphosphotidylinositol (GPI) anchor additional signal at its N- and C-terminuses, respectively. Potential N-glycosylation sites and domains involving cell adhesion were also identified in MoFLP1. Sequence analysis and subcellular localization by the expression of MoFLP1-GFP fusion construct in M. oryzae indicated that the MoFLP1 protein is probably localized on the vacuole membrane. Two MoFLP1 null mutants generated by targeted gene disruption exhibited marked reduction of conidiation, conidial adhesion, appressorium turgor, and pathogenicity. Our results indicate that fasciclin proteins play important roles in fungal development and pathogenicity in M. oryzae. Project supported by the National Natural Science Foundation of China (No. 30870101) and the Public Welfare Profession (Agriculture) Research Project (No. 200803008), China     

A CHASE domain containing protein kinaseOsCRL4, represents a newAtCRE1-like gene family in rice

AtCRE1 is known to be a cytokinin receptor inArabidopsis. TheAtCRE1 protein contains CHASE domain at the N-terminal part, followed by a transmitter (histidine kinase) domain and two receiver domains. The N-terminal CHASE domain ofAtCRE1 contains putative recognition sites for cytokinin. Five CHASE domains containing proteins were found in rice,osCRL1a,OsCRL1b,OsCRL2,OsCRL3, andOsCRL4.OsCRL1a,OsCRL1b,OsCRL2 andOsCRL3 contain the four domains existing inCRE1, whereasOsCRL4 only contains the CHASE domain and a putative Ser/Thr protein kinase domain. The authors cloned the encoding geneOsCRL4 and found that it represents a new member of the cytokinin receptor protein in rice. Project supported by the National Natural Science Foundation of China     

Purification and relationship with gastric disease of a 130 kDa (CagA) protein of<Emphasis Type="Italic">Helicobacter pylori</Emphasis>

Objective: The aims of this research were to purify and identify the 130 kDa (CagA) protein ofH. pylori clinical isolate HP97002 and evaluate the relationships between the purified 130 kDa (CagA) protein and gastric diseases. Methods: The procedure for isolating the protein included 6 mol/L guanidine extract, size exclusion chromatography and elusion from gel. Sera of 68 patients with gastric diseases (44 with chronic gastritis, 15 with atrophic gastritis, 7 with peptic ulcer disease, 2 with gastric cancer) were obtained, and the serological response to CagA was studied by Western-blot using the purified protein. Results: The purified protein was 130 kDa and preserved good antigenicity and revealed basic isoelectric point about of 8.1. Among 68 sera, 43 sera could recognize the purified protein associated with chronic gastritis 47.7% (21/44), atrophic gastritis 86.7% (13/15), peptic ulcer disease 100% (7/7), gastric cancer 100% (2/2). Compared with each other, the difference was significant (χ²=13.327,P=0.004), and 130 kDa (CagA) protein was associated with severe gastric diseases (r _s=0.442,P=0.001). Conclusion: The 130 kDa (CagA) protein was associated with severe gastric diseases. Project supported by the China Medical Board (96-628) and Zhejiang Province Hygiene Bureau (2000 A055)     

Different responses of two Mosla species to potassium limitation in relation to acid rain deposition

The increasingly serious problem of acid rain is leading to increased potassium (K) loss from soils, and in our field investigation, we found that even congenerically relative Mosla species show different tolerance to K-deficiency. A hydroponic study was conducted on the growth of two Mosla species and their morphological, physiological and stoichiometric traits in response to limited (0.35 mmol K/L), normal (3.25 mmol K/L) and excessive (6.50 mmol K/L) K concentrations. Mosla hangchowensis is an endangered plant, whereas Mosla dianthera a widespread weed. In the case of M. hangchowensis, in comparison with normal K concentration, K-limitation induced a significant reduction in net photosynthetic rate (P _n), soluble protein content, and superoxide dismutase (SOD) activity, but an increase in malondialdehyde (MDA) concentration. However, leaf mass ratio (LMR) and root mass ratio (RMR) were changed little by K-limitation. In contrast, for M. dianthera, K-limitation had little effect on P _n, soluble protein content, SOD activity, and MDA concentration, but increased LMR and RMR. Critical values of N (nitrogen): K and K:P (phosphorus) ratios in the shoots indicated that limitation in acquiring K occurred under K-limited conditions for M. hangchowensis but not for M. dianthera. We found that low K content in natural habitats was a restrictive factor in the growth and distribution of M. hangchowensis, and soil K-deficiency caused by acid rain worsened the situation of M. hangchowensis, while M. dianthera could well acclimate to the increasing K-deficiency. We suggest that controlling the acid rain and applying K fertilizers may be an effective way to rescue the endangered M. hangchowensis. Project supported by the National Natural Science Foundation of China (No. 30570113) and the Research Fund for the Doctoral Program of Higher Education (No. 20060335008), China     

Determination of the genus-specific antigens in outer membrane proteins from the strains of<Emphasis Type="Italic">Leptospira interrogans</Emphasis> and<Emphasis Type="Italic">Leptospira biflexa</Emphasis> with different virulence

Objective: To determine the existence of genus-specific antigens in outer membrane proteins (OMPs) ofleptospira with different virulence. Methods: Microscope agglutination test (MAT) was applied to detect the agglutination between commercial rabbit antiserum against leptospiral genus-specific TR/Patoc I antigen and 17 strains ofLeptospira interrongans belonging to 15 serogroups and 2 strains ofLeptospira biflexa belonging to 2 serogroups. The outer envelopes (OEs) ofLinterrogans serogroupIcterohaemorrhagiae serovarlai strainlai (56601) with strong virulence and serogroupPomona serovarpomona strainLuo (56608) with low virulence, andL. biflexa serogroupSemaranga serovarpatoc strainPatoc I without virulence were prepared by using the method reported in Auranet al. (1972). OMPs in the OEs were obtained by treatment with sodium deoxycholate. SDS-PAGE and western blot were used for analyzing the features of the OMPs on electrophoretic pattern and the immunoreactivity to the antiserum against TR/Patoc I antigen, respectively. Results: All the tested strains belonging to different leptospiral serogroups agglutinated to the antiserum against leptospiral genus-specific TR/Patoc I antigen with agglutination titers ranging from 1:256–1:512. A similar SDS-PAGE pattern of the OMPs from the three strains ofleptospira with different virulence was shown and the molecular weight of a major protein fragment in the OMPs was found to be approximately 60 Kda. A positive protein fragment with approximately 32 KDa confirmed by Western blot, was able to react with the antiserum against leptospiral genus-specific TR/Patoc I antigen, and was found in each the OMPs of the three stains ofleptospira. Conclusion: There are genus-specific antigens on the surface ofL.interrogans andL.biflexa. The OMP with molecular weight of 32 KDa may be one of the genus-specific protein antigens ofleptospira. Project (No. 39970678) supported by the National Natural Science Foundation of China     

Survival of the biocontrol agents Brevibacillus brevis ZJY-1 and Bacillus subtilis ZJY-116 on the spikes of barley in the field

Fusarium head blight (FHB) caused byFusarium graminearum is a devastating disease that results in extensive yield losses to wheat and barley. A green fluorescent protein (GFP) expressing plasmid pRP22-GFP was constructed for monitoring the colonization of two biocontrol agents,Brevibacillus brevis ZJY-116, on the spikes of barley and their effect on suppression of FHB. Survival and colonization of theBrevibacillus brevis ZJY-1 andBacillus subtilis ZJY-116 strains on spikes of barley were observed by tracking the bacterial transformants with GFP expression. Our field study revealed that plasmid pRP22-GFP was stably maintained in the bacterial strains without selective pressure. The retrieved GFP-tagged strains showed that the bacterial population fluctuation accorded with that of the rain events. Furthermore, both biocontrol strains gave significant protection against FHB on spikes of barley in fields. The greater suppression of barley FHB disease was resulted from the treatment of barley spikes with biocontrol agents before inoculation withF. graminearum. Project supported by the National Natural Science Foundation of China (No. 30230250) and Science and Technology Committee of Zhejiang Province (No. 2003C22029), China     

Evaluation of the dust and methanol extracts of Garcinia kolae for the control of Callosobruchus maculatus (F.) and Sitophilus zeamais (Mots)

Insecticidal effects of different doses of the dust and methanol extracts of Garcinia kolae on Collosobruchus maculatus and Sitophilus zeamais were tested. The dust had no significant effect on the two insects; none of them died even at 3 d after treatment. The methanol extracts, however, had rapid lethal effects on both C. maculatus and S. zeamais. The mortality of C. maculatus by the lowest concentration of methanol extracts ranged from 95%∼100% whereas in S. zeamais, the mortality ranged from 87.5%∼100% and 70%∼100% in concentrations of 1 g extract+3 ml methanol and 1 g extract+5 ml methanol, respectively, from 24 to 48 h. The least concentration of 1 g extract+15 ml methanol had no significant lethal effect on Sitophilus zeamais.      

A New Potent Inhibitor of Thrombin from the Leech Haemendipsa Yanyuanensis

1　Ｉｎｔｒｏｄｕｃｔｉｏｎ　Ｔｈｒｏｍｂｕｓｄｉｓｅａｓｅｏｆｈｅａｒｔａｎｄｂｒａｉｎｂｌｏｏｄｖｅｓｓｅｌｈａｓｂｅｃｏｍｅａｋｉｌｌｅｒｏｆｍａｎ .Ｂｅｃａｕｓｅｏｆｔｈｅｓｉｄｅｅｆｆｅｃｔｓｏｆｈａｅｍｏｒｒｈａｇｅｏｒｔｈｅｓｈｏｒｔｈａｌｆ ｌｉｆｅｉｎｖｉｖｏ ,ｔｈｅｐｒｅ ｓｅｎｔａｎｔｉｃｏａｇｕｌａｎｔｄｒｕｇｓａｒｅｕｎｓａｔｉｓｆｉｅｄ ,ａｎｄｓｅａｒｃｈ ｉｎｇｆｏｒｎｅｗａｎｔｉｃｏａｇｕｌａｎｔｓｗｉｔｈｈｉｇｈｅｒｅｆｆｉｃｉｅｎｔａｎｄｍｏｒｅｓｔａｂｉｌｅｈａ…     

李大钊《我的马克思主义观》新探——兼谈《新青年》第6卷各号的出版时间

《我的马克思主义观》是李大钊成为马克思主义者的标志,是五四新文化运动的重要转折点,也是马克思主义在中国开始广泛传播的重要标志。有关论著对它的研究和提及很多,《李大钊全集》的编注者对其注解也较多。进一步研究起来,该文中需要探究的问题仍有不少。总起来看,李大钊的《我的马克思主义观》以日本社会主义学者河上肇的《马克思的社会主义理论体系》为重要蓝本,其关于马克思主义三个组成部分与一条“金线”的核心论断也出于河上肇此文;写作和定稿时间,应为1919年7月底到8月上旬,而不可能在1919年5月之前。     

Deletion analysis of SMN1 and NAIP genes in southern Chinese children with spinal muscular atrophy

Spinal muscular atrophy (SMA) is a disorder characterized by degeneration of lower motor neurons and occasionally bulbar motor neurons leading to progressive limb and trunk paralysis as well as muscular atrophy. Three types of SMA are recognized depending on the age of onset, the maximum muscular activity achieved, and survivorship: SMA1, SMA2, and SMA3. The survival of motor neuron (SMN) gene has been identified as an SMA determining gene, whereas the neuronal apoptosis inhibitory protein (NAIP) gene is considered to be a modifying factor of the severity of SMA. The main objective of this study was to analyze the deletion of SMN1 and NAIP genes in southern Chinese children with SMA. Here, polymerase chain reaction (PCR) combined with restriction fragment length polymorphism (RFLP) was performed to detect the deletion of both exon 7 and exon 8 of SMN1 and exon 5 of NAIP in 62 southern Chinese children with strongly suspected clinical symptoms of SMA. All the 32 SMA1 patients and 76% (13/17) of SMA2 patients showed homozygous deletions for exon 7 and exon 8, and all the 13 SMA3 patients showed single deletion of SMN1 exon 7 along with 24% (4/17) of SMA2 patients. Eleven out of 32 (34%) SMA1 patients showed NAIP deletion, and none of SMA2 and SMA3 patients was found to have NAIP deletion. The findings of homozygous deletions of exon 7 and/or exon 8 of SMN1 gene confirmed the diagnosis of SMA, and suggested that the deletion of SMN1 exon 7 is a major cause of SMA in southern Chinese children, and that the NAIP gene may be a modifying factor for disease severity of SMA1. The molecular diagnosis system based on PCR-RFLP analysis can conveniently be applied in the clinical testing, genetic counseling, prenatal diagnosis and preimplantation genetic diagnosis of SMA. Project supported by the National Natural Science Foundation of China (No. J0710043), and the Natural Science Foundation of Zhejiang Province (No. 2007C33049), China     

Expression and purification of bioactive high-purity human midkine in Escherichia coli

Midkine is a heparin-binding growth factor, which plays important roles in the regulation of cell growth and differentiation. The non-tagged recombinant human midkine (rhMK) is therefore required to facilitate its functional studies of this important growth factor. In the present work, rhMK was expressed in Escherichia coli (E. coli) BL21 (DE3). The expression of midkine was efficiently induced by isopropyl-β-D-thiogalactopyranoside (IPTG). After sonication, midkine was recovered in an insoluble form, and was dissolved in guanidine hydrochloride buffer. Renaturation of the denatured protein was carried out in the defined protein refolding buffer, and the refolded protein was purified using S-Sepharose ion-exchange chromatography. The final preparation of the rhMK was greater than 98% pure as measured by sodium dodecylsulfate-polyacrylamid gel electrophoresis (SDS-PAGE) and reverse phase high performance liquid chromatography (RP-HPLC). The purified rhMK enhanced the proliferation of NIH3T3 cells. Project supported in part by the Hi-Tech Research and Development Program (863) of China (No. 2007AA02Z149) and the Science and Technology Commission of Shanghai Municipality (No. 075407071), China     

Influence of fasting on muscle composition and antioxidant defenses of market-size Sparus macrocephalus

The study was conducted to investigate fasting effects on flesh composition and antioxidant defenses of market-size Sparus macrocephalus. Two hundred fish (main initial weight 580 g) were divided into two groups (control and fasted) and reared in 6 cages. After two weeks of adaptation, group I fasted for 28 d; group II was fed normally as a control. In 3, 7, 14, 21 and 28 d, 6 fish per group were sampled for proximate flesh composition, liver antioxidant enzyme activities and malondialdehyde flesh content analyses. In fasted fish, the reduction of lipid content in muscle occurred after day 3, and, compared to controls, the content of protein decreased from day 14, the activities of liver antioxidative enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPX) increased from day 3, and flesh malondialdehyde levels increased from day 21. Flesh fat reduction shows that fasting may be used as a technique to reduce flesh lipid content in Sparus macrocephalus. However, considering flesh protein loss and the subsequent oxidative stress, the fasting technique should be used with precautions. Project (No. 2006C12098) supported by the Science and Technology Department of Zhejiang Province, China     

Protective effect of DNA-mediated immunization with liposome-encapsulated GRA4 against infection of Toxoplasma gondii

The dense granule protein 4 (GRA4) is a granular protein from Toxoplasma gondii, and is a candidate for vaccination against this parasite. In this study, the plasmid pcDNA3.1-GRA4 (pGRA4), encoding for the GRA4 antigen, was incorporated by the dehydration-rehydration method into liposomes composed of 16 mmol/L egg phosphatidylcholine (PC), 8 mmol/L dioleoyl phosphatidylethanolamine (DOPE), and 4 mmol/L 1,2-diodeoyl-3-(trimethylammonium) propane (DOTAP). C57BL/6 mice and BALB/c mice were immunized intramuscularly three times with liposome-encapsulated pGRA4 to determine whether DNA immunization could elicit a protective immune response to T. gondii. Enzyme-linked immunosorbent assay (ELISA) of sera from immunized mice showed that liposome-encapsulated pGRA4 generated high levels of IgG antibodies to GRA4. Production of primary interferon (IFN)-γ and interleukin (IL)-2 in GRA4-stimulated splenocytes from vaccinated mice suggested a modulated Th1-type response. 72.7% of C57BL/6 mice immunized with liposome-encapsulated pGRA4 survived the challenge with 80 tissue cysts of ME49 strain, whereas C57BL/6 mice immunized with pGRA4 had only a survival rate of 54.5%. When immunized BALB/c mice were intraperitoneally challenged with 10³ tachyzoites of the highly virulent RH strain, the survival time of mice immunized with liposome-encapsulated pGRA4 was markedly longer than that of other groups. Our observations show that liposome-encapsulated pGRA4 enhanced the protective effect against infection of T. gondii. Project supported by the Science Foundation of the Health Bureau of Zhejiang Province, China (Nos. 2003QN003 and 2005A001) and the Science Foundation of the Science and Technology Department of Zhejiang Province, China (No. 2006C13022)     

Review：BRCA1/2 associated hereditary breast cancer

Breast cancer is one of the leading causes of death in women today. Some of the patients are hereditary, with a large proportion characterized by mutation in BRCA1 and/or BRCA2 genes. In this review, we provide an overview of these two genes, focusing on their relationship with hereditary breast cancers. BRCA1/2 associated hereditary breast cancers have unique features that differ from the general breast cancers, including alterations in cellular molecules, pathological bases, biological behavior, and a different prevention strategy. But the outcome of BRCA1/2 associated hereditary breast cancers still remains controversial; further studies are needed to elucidate the nature of BRCA1/2 associated hereditary breast cancers. Project supported by the National Natural Science Foundation of China (No. 30772510) and the Joint Program of Ministry of Health and Zhejiang Provincal Government of China (No. WKJ2006-2-008)     

Microbial control of diamondback moth,Plutella xylostella L. (Lepidoptera: Yponomeutidae) using bacteria (Xenorhabdus nematophila) and its metabolites from the entomopathogenic nematode Steinernema carpocapsae

Cells and cell-free solutions of the culture filtrate of the bacterial symbiont, Xenorhabdus nematophila taken from the entomopathogenic nematode Steinernema carpocapsae in aqueous broth suspensions were lethal to larvae of the diamondback moth Plutella xylostella. Their application on leaves of Chinese cabbage indicated that the cells can penetrate into the insects in the absence of the nematode vector. Cell-free solutions containing metabolites were also proved as effective as bacterial cells suspension. The application of aqueous suspensions of cells of X. nematophila or solutions containing its toxic metabolites to the leaves represents a possible new strategy for controlling insect pests on foliage.     

严羽《诗辩》文本顺序问题补议

《沧浪诗话》今传"通行本"与"玉屑本"两种版本,其中《诗辩》的文本顺序存在较大差异。文章在综述已有研究成果的基础上,进一步就《诗辩》的文本顺序问题提出四点补论:其一,《沧浪诗话》原非"诗话体"著作,《诗辩》当为"辩"体论说文,而《诗人玉屑》作为诗话著作摘录《诗辩》,并不承担保存其原貌的责任;其二,若依"玉屑本",则《诗辩》开篇即蹈袭北宋《潜溪诗眼》旧话;其三,从两个"识"字可以看出被"玉屑本"腰斩为两段的文字原本存在紧密的逻辑承接关系,且"通行本"在行文脉络上更加明晰;其四,论法论品相关文字或为"通行本"编校者据"玉屑本"补入,是以此两段文字依"玉屑本"当更为合理。     

Genetic linkage map ofBrassica campestris L. using AFLP and RAPD markers

A genetic linkage map comprised of 131 loci was constructed with an F₂ population derived from an inter-subspecific cross betweenBrassica campestris L. ssp.chinensis cv. ‘aijiaohuang’ and ssp.rapifera cv. ‘qisihai’. The genetic map included 93 RAPD loci, 36 AFLP loci and 2 morphological loci organized into 10 main linkage groups (LGs) and 2 small groups, covering 1810.9cM with average distance between adjacent markers being approximately 13.8cM. The map is suitable for identification of molecular markers linked to important agronomic traits, QTL analysis, and even for marker-assisted selection in breeding programs of Chinese cabbage and turnip. Project (No. 39870509) supported by the National Natural Science Foundation of China     

Electronic structure and magnetism ofRMn6Sn6 (R=Tb, Dy)

This article reports first-principles band structure calculations forRMn₆Sn₆ (R=Tb, Dy). The calculation uses the linear muffin-tin orbitals (LMTO) method in the atomic-sphere-approximation (ASA), and yields results showing that both TbMn₆Sn₆ and DyMn₆Sn₆ are ferrimagnetic compounds with antiparallel aligned moments ofR and Mn atoms. In this research the 4f states ofR atoms are treated as localized states, i.e., the hybridization of 4f states with other valence electrons is neglected. The moments of Mn in both compounds were determined to be 2.43 μ_B and 2.38 μ_B, respectively. The considerably small additional moments for Mn from the spin-orbit coupling indicates that the spin-orbital coupling is not dominated for Mn atoms. The total moments of Tb and Dy atoms are 10.28 μ_B and 11.20 μ_B. All the calculation findings accorded well with experimental results. Project supported by the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry of China.