Study of serum<Emphasis Type="Italic">Helicobacter pylori</Emphasis> soluble antigen

Objective: to explore a new serological method for detectingHelicobacter pylori (H. pylori) infection. Methods: Serum soluble antigen ofH. pylori was detected by using avidin-biotin ELISA technique to evaluate the status ofH. pylori infection and for comparison with rapid urease test (RUT), histologic examination and serology. Results: The sensitivity, specificity, positive predictive value and negative predictive value were 77.46%, 91.07%, 91.67% and 76.12%, respectively. The prevalence rate of serumH. pylori soluble antigen in 138 patients undergoing endoscopy was similar to the rate obtained by¹⁴C-UBT methods (P>0.05). Conclusions: The detection of serumH. pylori soluble antigen(HpSAg) could be used as a new serological method which is accurate, and convenient, not affected by the memorizing reaction of serum antibody; is more sensitive, more specific and suitable for clinical diagnosis, and evaluation of eradication and for follow-up ofH. pylori as well as for detection in children and pregnant women. Project supported by Zhejiang Provincial Health Bureau (No. 2000A118), China     

Exact failure probability formula for strict consecutive-k-out-of-n:F systems with homogeneous Markov dependence

An exact formula is presented for calculating the failure probability of strict consecutive-k-out-of-n:F systems with homogeneous Markov dependence Project supported by the National Natural Science Foundation of China (19271084)     

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     

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)     

Study on the serum levels of soluble intercellular adhesion molecule-1 (sICAM-1) in patients withHelicobacter pylori infection

Objective: To evaluate the interaction between serum levels of soluble intercellular adhesion molecule-1 (sICAM-1) andHelicobacter pylori (H. pylori) infection in patients with chronic gastritis and peptic ulcer. Methods: The serum levels of sICAM-1 in 205 patients with chronic gastric diseases were detected by ELISA method and the status ofH. pylori was determined by histologic examination, RUT,¹⁴C-UBT, and serology. The sera obtained from 18 healthy volunteers served as controls. Results: The serum levels of sICAM-1 were significantly higher in patients withH. pylori positive than those of H. pylori negative (889.43±32.52 ng/ml vs. 747.07±30.45 ng/ml,P<0.05). The serum levels of sICAM-1 in patients with mild, moderate and severe infection ofH. pylori were 841.68±72.36 ng/ml, 905.43±37.59 ng/ml and 1012.54±49.34 ng/ml, respectively (P<0.05). The serum levels of sICAM-1, proved to be significantly correlated with the density ofH. pylori colonization in gastric mucosa (r _s=0.316,P<0.001). The serum levels of sICAM-1 in patients with chronic gastritis and peptic ulcer were significantly higher than those in healthy controls (P<0.05). Conclusions: These results indicated thatH. pylori infection up-regulates the expression of sICAM-1.     

Distribution of<Emphasis Type="Italic">H. pylori</Emphasis> antigens in gastric mucosa and its significance

Objective: To investigate the distribution ofH. pylori antigens in the gastric mucosa in patients withH. pylori infection, and the relationship between the distribution and gastric cancer. Methods: Of 112 patients confirmed by pathological study to have chronic superficial gastritis, precancerous changes (chronic atrophic gastritis, intestinal metaplasia or atypical hyperplasia) and gastric cancer, 28 wereH. pylori negative and 84 wereH. pylori positive.H. pylori antigens in the gastric mucosa were detected by immunohistochemistry. Results: TheH. pylori positive group, comprised 12 of 22 (50.0%) in the chronic superficial gastritis group, 22 of 25 (88.0%) in the precancerous changes group and 13 of 35 (37.1%) in the gastric cancer group. The positive rates ofH. pylori antigens in the cytoplasm progressively increased, respectively at 0.0% (0/12), 63.6% (14/22) and 84.6% (11/13) for the same groups (χ ²=19.76,P=0.000);H. pylori antigens were located in the mucus layer and above the neck of the mucosal gland in 9 of 12 (75.0%) cases with chronic superficial gastritis, at the neck of the mucosal gland and the isthmus in 12 of 22 (54.5%) cases with precancerous changes, below the isthmus in 9 of 13 (69.2%) cases with gastric cancer (χ ²=25.30,P=0.000). In theH. pylori negative group, noH. pylori antigen was observed. Conclusion: With the progression of chronic superficial gastritis→precancerous changes→gastric cancer,H. pylori antigens progressively migrated from the outer part to the inner part of the cell, and from the superficial to the deep gastric mucosa.     

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     

Study on the expression and mutation of human telomeric repeat binding factor ( hTRF1 ) in 10 malignant hematopoietic cell lines

Objective: Detecting the expression and mutation of human telomeric repeat binding factor (hTRF1) in 10 malignant hematopoietic cell line cells on the base of determining its genomic structure and its four pseudogenes to clarify if hTRF1 mutation is one of the factors of the activation of telomerase. Methods: hTRF1cDNA sequences were obtained from GenBank, its genome structure and pseudogenes were forecasted by BLAST and other biology information programs and then testified by sequencing. Real-time RT-PCR was used to detect the expression of hTRF1mRNA in 10 cell line cells, including myelogenous leukemia cell lines K562, HL-60, U-937, NB4, THP-1, HEL and Dami; lymphoblastic leukemia cell lines 6T-CEM, Jurkat and Raji. Telomerase activities of cells were detected by using telomeric repeat amplification (TRAP)-ELISA protocol. PCR and sequencing were used to detect mutation of each exon of hTRF1 in 10 cell line cells. Results: hTRF1 gene, mapped to 8q13, was divided into 10 exons and spans 38.6 kb. Four processed pseudogenes of hTRF1 located on chromosome 13, 18, 21 and X respectively, was named as ψhTRF1-13, ψhTRF1-18, ψhTRF1-21 and ψhTRF1-X respectively. All cell line cells showed positive telomerase activity. The expression of hTRF1 was significantly lower in malignant hematopoietic cell lines cells (0.0338, 0.0108:_0.0749) than in normal mononuclear cells (0.0493, 0.0369:_0.128) (P=0.004). But no significant mutation was found in all exons of hTRF1 in 10 cell line cells. Four variants were found in part of intron 1, 2 and 8 of hTRF1. Their infection on gene function is unknown and needs further studies. Conclusion: hTRF1 mutation is probably not one of the main factors for telomerase activation in malignant hematopoietic disease. Project supported by the National Basic Research Program (973) of China (No. 2002CB713700) and the National Natural Science Foundation of China (No. 39870339)     

Effects of jinlu, an anti-juvenile hormone on the growth, ultra-structure of the corpora allata and prothoracic gland of silkworm,Bombyx mori L

The 4th instar larvae of the silkworm,Bombyx mori L, when treated with anti-juvenile hormone (Jinlu) had its larval period extended by 2 days and the total larval period shortened by about 4 days. The conversion ratio of tetramolters into trimolters was 100%. But anti-juvenile hormone administration to the 5th instar larvae lengthened the silkworm age by one day. When anti-juvenile hormone was administered, we could find many neurosecretory granules of the brain transferred to the cells of the corpora allata, but there was little endoplasmic reticulum. In the prothoracic gland, the micropile edge was clear and there were large nucleoli, mitochondria and endoplasmic reticulum. This study was carried out to show that anti-JH compound inhibits the secretion of Juvenile hormone in silkwormBombyx mori L. The investigation revealed that the antijuvenile hormone inhibited the secretion of corpora allata and initiated the activity of the prothoracic gland. Project supported by the Government of China and Ministry of Human Resource Development, Government of India for Bilateral exchange program. Post-Doctoral Scholar in College of Animal Sciences of Zhejiang University now, from Sri Padmavathi Women’s University, India.     

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     

How is nature asymmetric?

This two-part article considers certain fundamental symmetries of nature, namely the discrete symmetries of parity (P), charge conjugation (C) and time reversal (T), and their possible violation. Recent experimental results are discussed in some depth. In this second part, we discussCP andT violation and arrive at a synthesis.     

A Note on Vulnerability in Graphs of Diameter Five （ Ⅱ ）

The line persistence of a graph G, Pt （ G ） is the minimum number of lines which must be removed to increase the diameter of G. In Ref. [7] （J. Shanghai Univ., 2003,7（4）：352-357）, we gave a characterization of graphs of diameter five with ρ1 （ G ）≥2. In this paper we will show that each of the 8 special graphs Xi （ i = 1,2,3,4,5,6,7,8） listed in condition （2） of Theorem 1 in Ref. [7] can not be deleted. Therefore the results we obtained in Ref. [7] can not in general be improved.     

Studies on the management of root-knot nematode, Meloidogyne incognita -wilt fungus, Fusarium oxysporum disease complex of green gram, Vigna radiata cv ML-1108

Studies were conducted under pot conditions to determine the comparative efficacy of carbofuran at 1 mg a.i./kg soil, bavistin at 1 mg a.i./kg soil, neem (Azadirachta indica) seed powder at 50 mg/kg soil, green mould (Trichoderma harzianum) at 50.0 ml/kg soil, rhizobacteria (Pseudomonas fluorescens) at 50.0 ml/kg soil against root-knot nematode,Meloidogyne incognita-wilt fungus,Fusarium oxysporum disease complex on green gram,Vigna radiata cv ML-1108. All the treatments significantly improved the growth of the plants as compared to untreated inoculated plants. Analysis of data showed that carbofuran andA. indica seed powder increased plant growth and yield significantly more in comparison to bavistin andP. fluorescens. Carbofuran was highly effective against nematode, bavistin against fungus,A. indica seed powder against both the pathogens and both the bioagents were moderately effective against both the pathogens.     

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。     

Proteome changes in the plasma of Papilio xuthus (Lepidoptera: Papilionidae): effect of parasitization by the endoparasitic wasp Pteromalus puparum (Hymenoptera: Pteromalidae)

Although the biochemical dissection of parasitoid-host interactions is becoming well characterized, the molecular knowledge concerning them is minimal. In order to understand the molecular bases of the host immune response to parasitoid attack, we explored the response of Papilio xuthus parasitized by the endoparasitic wasp Pteromalus puparum using proteomic approach. By examining the differential expression of plasma proteins in the parasitized and unparasitized host pupae by two-dimensional (2D) electrophoresis, 16 proteins were found to vary in relation to parasitization compared with unparasitized control samples. All of them were submitted to identification by mass spectrometry coupled with a database search. The modulated proteins were found to fall into the following functional groups: humoral or cellular immunity, detoxification, energy metabolism, and others. This study contributes insights into the molecular mechanism of the relationships between parasitoids and their host insects. Project supported by the National Basic Research Program (973) of China (No. 2006CB102005), the National Natural Science Foundation of China (Nos. 30571251 and 30170626), the Program for New Century Excellent Talents in University of the Ministry of Education of China (No. NCET-05-0513), and the Innovation Research Team Program of the Ministry of Education of China (No. IRT0535)     

Empowering Adolescent Readers: Intertextuality in Three Novels by David Almond

In Skellig, Kit’s Wilderness, and Clay, David Almond employs various types of intertextuality to enrich his narratives. Through the use of allusion, adaptation, collage, and mise-en-abyme, he encourages his adolescent readers to seek out precursor texts and to consider the interrelationships between these texts and his own. By so doing, he demonstrates the respect he has for his readers and empowers them to become active makers of meaning.

《现代汉语词典》6版量词修订浅析

《现代汉语词典》6版对5版作了进一步修订,其中对量词的修订主要体现在对已有量词的修订、新增量词、删除量词等三个方面。     

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     

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     

Active changes of lignification-related enzymes in pepper response to Glomus intraradices and/or Phytophthora capsici

The activities of enzymes responsible for lignification in pepper, pre-inoculation with arbuscular mycorrhizal (AM) fungus ofGlomus intraradices and/or infection with pathogenic strain ofPhytophthora capsici, and the biological control effect ofG. intraradices on Phytophthora blight in pepper were investigated. The experiment was carried out with four treatments: (1) plants pre-inoculated withG. intraradices (Gi), (2) plants pre-inoculated withG. intraradices and then infected withP. capsici (Gi+Pc). (3) plants infected withP. capsici (Pc), and (4) plants without any of the two microorganisms (C). Mycorrhizal colonization rate was reduced by about 10% in pathogen challenged plants. Root mortality caused by infection ofP. capsici was completely eliminated by pre-inoculation with antagonisticG. intraradices. On the ninth day after pathogen infection, Peroxidase (POD) activity increased by 116.9% in Pc-treated roots but by only 21.2% in Gi+Pc-treated roots, compared with the control, respectively. Polyphenol oxidase (PPO) and Phenylalanine ammonia-lyase (PAL) activities gradually increased during the first 3 d and dramatically decreased in Pc-treated roots but slightly decreased in Gi+Pc-treated roots, respectively. On the ninth day after pathogen infection, PPO and PAL decreased by 62.8% and 73.9% in Pc-treated roots but by only 19.8% and 19.5% in Gi+Pc-treated roots, compared with the control, respectively. Three major POD isozymes (45000, 53000 and 114000) were present in Pc-treated roots, while two major bands (53000 and 114000) and one minor band (45000) were present in spectra of Gi+Pc-treated roots, the 45000 POD isozyme was significantly suppressed byG. intraradices, suggesting that the 45000 POD isozyme was induced by the pathogen infection but not induced by the antagonisticG. intraradices. A 60000 PPO isozyme was induced in Pc-treated roots but not induced in Gi+Pc-treated roots. All these results showed the inoculation of antagonisticG. intraradices alleviates root mortality, activates changes of lignification-related enzymes and induces some of the isozymes in pepper plants infected byP. capsici. The results suggested thatG. intraradices is a potentially effective protection agent againstP. capsici. Project supported by Korea Science and Engineering Foundation (KOSEF) through the Agricultural Plants Stress Research Center (APSRC) at Chonnam National University, Korea