Purification and relationship with gastric disease of a 130 kDa (CagA) protein of Helicobacter pylori

Objective: The aims of this research were to purify and identify the 130 kDa (CagA) protein of H. 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 (rs=0.442, P=0.001). Conclusion: The 130 kDa (CagA) protein was associated with severe gastric diseases.     

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.     

Morphological and pathologic changes of experimental chronic atrophic gastritis （CAG） and the regulating mechanism of protein expression in rats

INTRODUCTION In 1998 gastrointestinal pathologists reached aconsensus on the definition of chronic atrophic gas- tritis (CAG), which was described as programmed loss of gastric gland and/or replacement by intestinal glands in gastric mucosa. CAG was recognized to be closely related with development of gastric cancer and listed as precancerous lesions in this meeting (Genta, 1998). According to Correa (1992)’s cascade of gas-tric carcinogenesis, gastric cancer was believed to develop fr…     

Study on the serum levels of soluble intercellular adhesion molecule-1 (sICAM-1) in patients with Helicobacter pylori Infection

Objective: To evaluate the interaction between serum levels of soluble intercellular adhesion molecule-1 (sICAM-1) and Helicobacter 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 of H. 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 with H. 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 of H. 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 of H. pylori colonization in gastric mucosa (rs =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 that H. pylori infection up-regulates the expression of sICAM-1.     

Expression of three essential antioxidants of Helicobacter pylori in clinical isolates

Objective

Helicobacter pylori maintains long-term persistence in the host and combats oxidative stress via many antioxidant proteins, which are expected to be relevant to bacterial-associated gastric diseases. We aimed to investigate the expression of three essential antioxidants in H. pylori strains isolated from patients with different clinical outcomes. Methods: Forty H. pylori strains were isolated from endoscopic biopsy specimens of gastric mucosa from 13 patients with gastric cancer, 13 with peptic ulcer, and 14 with gastritis. The expression of thioredoxin 1 (Trx1), arginase (RocF), and alkyl hydroperoxide reductase (AhpC) in H. pylori was measured by real-time PCR. Comparisons among multiple sample sets were analyzed using a one-way ANOVA test. Pearson’s correlation test was used to assess relationships among multiple continuous variables.

Results

Trx1 expression of H. pylori in gastric cancer and peptic ulcer tissues was higher than that in tissues with gastritis. RocF expression of H. pylori in gastric cancer tissues was higher than that in tissues exhibiting peptic ulcer and gastritis. However, we did not find any differences in AhpC expression in samples from patients with different clinical outcomes. The expression of Trx1 and RocF had a positive, linear correlation. The expression of Trx1 and AhpC had a positive correlation without a linear trend. We found no correlation between the expression of RocF and AhpC.

Conclusions

Our observations indicate that the expression of Trx1 and RocF in H. pylori might be related to gastric carcinogenesis. In H. pylori, the expression of members of the antioxidant system may be correlated and relevant to gastric cancer.     

The relationship between ulcer recurrence andHelicobacter pylori: A prospective one-year follow-up study in China

To study the relationship between ulcer recurrence andHelicobacter Pylori. 147 patients with peptic ulcer were divided intoH. Pylori infected group andH. pylori uninfected group by rapid urease test serum antibody examination and histopathology of gastric biopsy specimens. We assigned all patients to receive dual oral therapy with omeprazole 40 mg/day for 4 weeks and amoxicillin 2000 mg/day for 2 weeks. To all patients, repeated gastroscope was performed at 8 weeks and one-year after beginning therapy to record whether the ulcer was cured. Endoscopic biopsy was also performed to determineH. pylori status. One year later,H. pylori positive patients had a higher ulcer recurrent rate thanH. pylori negative patients (23. 33% vs 2. 70%; P=0.001). Ulcer recurrence was closely related toH. pylori infection.H. pylori was the risk factor for ulcer relapse.     

718例纤维胃镜活检组织检查结果

本文通过718例纤维胃镜活检组织险查结果表明,胃病的发病情况慢性浅表性胃炎>慢性萎缩性胃炎>胃溃疡>胃息肉>糜烂性胃炎>胃癌,其中以慢性浅表性胃炎患病率最高占55%。胃镜与病理诊断符合率81.06%,男女比为3:1。浅表性胃炎的程度越重合并病变越多,对中度以上不典型增生的高危人群应定期检查,以便及时治疗。     

A bug that can dig a hole in the stomach!

Barry J Marshall and J Robin Warren from Australia received the Nobel Prize for their discovery of the role of the bacteriumHelicobacter pylori in gastritis and peptic ulcer. Their findings challenged the prevailing dogma about peptic ulcer. Thanks to their pioneering work, peptic ulcer is no longer a chronic, frequently disabling condition, but a disease that can be cured by a short regimen of antibiotics and acid secretion inhibitors.     

Gastric motility functional study based on electrical bioimpedance measurements and simultaneous electrogastrography

For some time now, the research on gastric motility and function has fallen behind in the amount of research on gastric endocrine, exocrine secretion, and gastric morphology. In this paper, a noninvasive method to study gastric motility was developed, taking bioimpedance measurements over the gastric area simultaneously with the electrogastrography (EGG). This is based on the concept of observing and analyzing simultaneously the intrinsic electrical gastric activity (basic electric rhythm) and the mechanical gastric activity. Additionally, preliminary clinical studies of healthy subjects and subjects with functional dyspepsia (FD) and gastritis were carried out. The impedance gastric motility (IGM) measurements of the healthy and FD subjects were compared, along with the studies of the FD subjects before treatment and after one week and three weeks of treatment. We also compared IGM measurements of healthy subjects and subjects with erosive gastritis, along with the studies of the subjects with erosive gastritis before treatment and after one week of treatment. Results show that FD subjects have poor gastric motility (P<0.01). After a week of treatment, the gastric motility of FD subjects was not yet improved although the EGG had returned to normal by this time. By three weeks of treatment, the regular IGM rhythm returned in FD subjects. There was a significant difference of IGM parameters between the gastritis and healthy subjects (P<0.05). The EGG rhythm of the gastritis subjects returned to normal at one week post-treatment, while IGM parameters showed a trend to improvement (P>0.05), These results suggest the possibility of clinic application of the proposed method.     

消化性溃疡与蛋白质

认为消化性溃疡（Pepticulcer）病的发生、治疗和预防与食物中蛋白质的缺乏有着极为密切的关系,技正了“已经误导我们将近一个世纪的‘无酸就无溃疡’（Noacid,Noulcer）的论点”,并且指出中和胃酸和抑制胃酸的药物不可能根治消化性溃疡,只有向食物中添加足够量的蛋白质或蛋白质降解产物才是治疗和预防消化性溃疡的根本之所在！     

幽门螺旋杆菌感染与耳鼻咽喉头颈外科疾病关系研究综述

幽门螺旋杆菌与慢性胃炎及消化性溃疡密切相关,但最近研究发现与某些耳鼻咽喉头颈外科疾病亦有一定关系。本文从阐述幽门螺旋杆菌的生物学性状、诊断技术、流行病学及抗幽门螺旋杆菌治疗入手,综述了幽门螺旋杆菌与耳鼻咽喉头颈外科疾病的关系,为从系统因素出发研究耳鼻咽喉头颈外科疾病提供新思路。     

Distribution of H.pylori antigens in gastric mucosa and its significance

Objective: To investigate the distribution of H. pylori antigens in the gastric mucosa in patients with H. 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 were H. pylori negative and 84 were H. pylori positive. H. pylori antigens in the gastric mucosa were detected by immunohistochemistry. Results: The H. 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 of H. 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 (χ2=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 (x2=25.30, P=0.000). In the H. pylori negative group, no H.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.     

Distribution of H.pylori antigens in gastric mucosa and its significance

INTRODUCTION Helicobacter pylori (H.pylori) was grouped as a class I carcinogen by the International Agency for Research on Cancer in 1994. A direct relation be-tween H.pylori infection and gastric carcinogenesis was demonstrated in 1998 in an experimental animal model (Watanabe et al., 1998). However, the role of H.pylori in human gastric carcinogenesis is sup-ported almost exclusively by epidemiological data and prospective histopathological studies. So far the mechanism of H.pylor…     

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     

Purification and characterization of a novel antifungal protein from Bacillus subtilis strain B29

An antifungal protein was isolated from a culture of Bacillus subtilis strain B29. The isolation procedure comprised ion exchange chromatography on diethylaminoethyl (DEAE)-52 cellulose and gel filtration chromatography on Bio-Gel® P-100. The protein was absorbed on DEAE-cellulose and Bio-Gel® P-100. The purified antifungal fraction was designated as B29I, with a molecular mass of 42.3 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), pI value 5.69 by isoelectric focusing (IEF)-PAGE, and 97.81% purity by high performance liquid chromatography (HPLC). B29I exhibited inhibitory activity on mycelial growth in Fusarium oxysporum, Rhizoctonia solani, Fusarium moniliforme, and Sclerotinia sclerotiorum. The 50% inhibitory concentrations (IC₅₀) of its antifungal activity toward Fusarium oxysporum and Rhizoctonia solani were 45 and 112 μmol/L, respectively. B29I also demonstrated an inhibitory effect on conidial spore germination of Fusarium oxysporum and suppression of germ-tube elongation, and induced distortion, tumescence, and rupture of a portion of the germinated spores.     

Cellular stress and redox activity proteins are involved in gastric carcinogenesis associated with <Emphasis Type="Italic">Helicobacter pylori</Emphasis> infection expressing high levels of thioredoxin-1

Helicobacter pylori infection is related to the development of gastric diseases. Our previous studies showed that high thioredoxin-1 (Trx1) expression in H. pylori can promote gastric carcinogenesis. To explore the underlying molecular mechanisms, we performed an isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomic analysis of stomach tissues from Mongolian gerbil infected with H. pylori expressing high and low Trx1. Differences in the profiles of the expressed proteins were analyzed by bioinformatics and verified using Western blot analysis. We found three candidate proteins, 14-3-3α/β, glutathione-S-transferase (GST), and heat shock protein 70 (HSP70), in high Trx1 tissues compared with low Trx1 tissues and concluded that cellular stress and redox activityrelated proteins were involved in the pathogenesis of gastric cancer associated with H. pylori Trx1.     

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)     

Cloning and characterization of a glucose 6-phosphate/phosphate translocator from<Emphasis Type="Italic">Oryza sativa</Emphasis>

Plastids of nongreen tissues import carbon as a source of biosynthetic pathways and energy, and glucose 6-phosphate is the preferred hexose phosphate taken up by nongreen plastids. A cDNA clone encoding glucose 6-phosphate/phosphate translocator (GPT) was isolated from a cDNA library of immature seeds of rice and named asOsGPT. The cDNA has one uninterrupted open reading frame encoding a 42 kDa polypeptide possessing transit peptide consisting of 70 amino acid residues. TheOsGPT gene maps on chromosome 8 of rice and is linked to the quantitative trait locus for 1000-grain weight. The expression ofOsGPT is mainly restricted to heterotrophic tissues. These results suggest that glucose 6-phosphate imported viaGPT can be used for starch biosynthesis in rice nongreen plastids. Project supported by National Natural Scienc Foundation of China (No.39830250) and Natural Science Foundation of Zhejiang Province (No.2A0106), China. The nucleotide sequence data will appear in the GenBank under accession number AF375053.     

Protective effects of Salvia miltiorrhizae on the hearts of rats with severe acute pancreatits or obstructive jaundice

Objective  To investigate the therapeutic effects and mechanisms of Salvia miltiorrhizae (Danshen) in the treatment of severe acute pancreatitis (SAP)- or obstructive jaundice (OJ)-induced heart injury. Methods  A total of 288 rats were used for SAP- (n=108) and OJ-associated (n=180) experiments. The rats were randomly divided into sham-operated, model control, and Salvia miltiorrhizae-treated groups. According to the difference of time points after operation, SAP rats in each group were subdivided into 3, 6 and 12 h subgroups (n=12), whereas OJ rats were subdivided into 7, 14, 21, and 28 d subgroups (n=15). At the corresponding time points after operation, the mortality rates of the rats, the contents of endotoxin and phospholipase A₂ (PLA₂) in blood, and pathological changes of the hearts were investigated. Results  The numbers of dead SAP and OJ rats in the treated groups declined as compared with those in the model control group, but not significantly (P>0.05). The contents of endotoxin (at 6 and 12 h in SAP rats and on 7, 14, 21, and 28 d in OJ rats, respectively) and PLA₂ (at 6 and 12 h in SAP rats and on 28 d in OJ rats, respectively) in the treated group were significantly lower than those in the model control group (P<0.01 and P<0.001, respectively). Besides, myocardial pathological injuries were mitigated in SAP and OJ rats. Conclusion  In this study, we found that Salvia miltiorrhizae improved myocardial pathological changes, reduced the content of PLA₂ in blood, and decreased the mortality rates of SAP and OJ rats, exerting protective effects on the hearts of the rats. Projected supported by the Technological Foundation Project of Traditional Chinese Medicine Science of Zhejiang Province (Nos. 2003C130 and 2004C142), the Grave Foundation Project for Technological and Development of Hangzhou City (No. 2003123B19), and the Intensive Foundation Project for Technology of Hangzhou City (No. 2004Z006), China