Different dysregulations of CYFIP1 and CYFIP2 in distinct types of dementia.

In humans, the cytoplasmic FMR1 interacting protein (CYFIP) family consists of two members, namely CYFIP1 and CYFIP2. Both CYFIP1 and CYFIP2 function in the WAVE regulatory complex (WRC), which regulates actin polymerization. Additionally, these two proteins form a posttranscriptional regulatory complex with the fragile X mental retardation protein (FMRP), which suppresses mRNA translation. Thus, CYFIP1 and CYFIP2 are important signalling regulators at synapses, and mutations in their genes are associated with neurodevelopmental and neuropsychiatric disorders, including intellectual disabilities. Moreover, dysregulation of the CYFIP protein family is involved in Alzheimer's disease (AD). However, the relevance of the CYFIP family in other dementias is largely unknown. Here, we compared CYFIP1/2 protein levels in the post-mortem hippocampus from patients with AD, dementia with Lewy bodies (DLB), vascular dementia (VaD) and frontotemporal lobar degeneration with TDP-43 pathology (FTLD-TDP). Consistent with previous findings, CYFIP2 was reduced in AD hippocampus. In DLB and VaD hippocampus, the protein level of CYFIP2 and CYFIP1 was unaltered. Finally, an increase in the protein level of both CYFIP1 and CYFIP2 was noted in FTLD-TDP hippocampus. These findings reveal that the protein levels of the CYFIP family is distinct in different types of dementia, suggesting that the pathogenesis of these neurodegenerative disorders has divergent impacts on hippocampal synaptic function.

Rapid turnover and short-term blooms of Escherichia coli in the human gut.

Escherichia coli (E. coli) is a common microorganism that is widely present in the environment and closely related to human health. The extent of E. coli presence in the human gut has been a subject of ongoing debate. Through whole-genome shotgun metagenomic sequencing, our study revealed that E. coli exists in the human body at a low abundance (average abundance 1.21%), with occasional short-term bursts leading to temporary increases in abundance, with the highest recorded at 50.91%. Further investigations into the factors contributing to these short-term blooms of E. coli showed significant variations in strain types and genomes within fecal samples collected from the same individuals at different time points. Evolutionary tree analysis indicated that samples from different individuals crossed, suggesting a change in the dominant E. coli strains within the human gut. Therefore, it can be inferred that E. coli in the human body are more likely to be transient bacteria rather than permanent residents in the gut. The rapid rate of turnover among months (87.5% within a month) and short-term blooms of E. coli in the human body can establish "latent infections" of nonpathogenic strains in healthy individuals while also posing a potential risk of introducing pathogenic strains, thereby impacting human health. In summary, our study revealed the variation in E. coli abundance and strains within the human gut, influenced by geographic area and temporal factors. These findings contribute to a better understanding of the relationship between E. coli, the gut microbiota, and human health. IMPORTANCE Escherichia coli (E. coli) is a microorganism closely linked to human health, and its presence in the human gut has been a topic of debate. Our study, using whole-genome shotgun metagenomic sequencing, revealed that E. coli exists at a low abundance in the human body, with occasional short-term bursts leading to temporary increases. Strain and genome variations were observed within fecal samples from the same individuals at different time points, suggesting transient rather than permanent residence of E. coli in the gut. The rapid turnover rate and short-term blooms of E. coli can establish latent infections while also posing a risk of introducing pathogenic strains. These findings enhance our understanding of the relationship between E. coli, the gut microbiota, and human health.

Genetic characteristics of Blastocystis sp. ST3 at the genome level in the Chinese population.

The gut microbiota comprises the collective genomes of microbial symbionts and is composed of bacteria, fungi, viruses, and protists within the gastrointestinal tract of a host. Although the literature associated with gut microbiota is increasing, studies on eukaryotes in the human gut are just beginning to surface. Blastocystis is one of the most common intestinal parasites of humans and animals and is estimated to colonise more than 1 billion people on a global scale. However, the understanding of the genetic characteristics of Blastocystis subtype (ST) at the genome level and its relationship with other members of the gut microbiota is still limited. In this study, by surveying the prevalence and genome characteristics of Blastocystis sp. ST3 in a Chinese population (prevalence % = 6.09%), the association of Blastocystis sp. ST3 with region and time and the structure of the resident gut bacterial population was clarified. We identified novel sequences (50 mitochondrial and 41 genome sequences) and determined their genetic diversity amongst strains within Blastocystis sp. ST3 (4.14 SNPs/kb). Furthermore, we found that colonisation of Blastocystis was strongly associated with increased bacterial richness and higher abundance of several anaerobes. Finally, we performed time series sampling on two Blastocystis-positive individuals and confirmed that Blastocystis could exist continually in the human gut microbiota and persist for a long time, even for 4 years.

Research on automatic classification and detection of chicken parts based on deep learning algorithm.

Accurate classification and identification of chicken parts are critical to improve the productivity and processing speed in poultry processing plants. However, the overlapping of chicken parts has an impact on the effectiveness of the identification process. To solve this issue, this study proposed a real-time classification and detection method for chicken parts, utilizing YOLOV4 deep learning. The method can identify segmented chicken parts on the assembly line in real time and accurately, thus improving the efficiency of poultry processing. First, 600 images containing multiple chicken part samples were collected to build a chicken part dataset after using the image broadening technique, and then the dataset was divided according to the 6:2:2 division principle, with 1200 images as the training set, 400 images as the test set, and 400 images as the validation set. Second, we utilized the single-stage target detector YOLO to predict and calculate the chicken part images, obtaining the categories and positions of the chicken leg, chicken wing, and chicken breast in the image. This allowed us to achieve real-time classification and detection of chicken parts. This approach enabled real-time and efficient classification and detection of chicken parts. Finally, the mean average precision (mAP) and the processing time per image were utilized as key metrics to evaluate the effectiveness of the model. In addition, four other target detection algorithms were introduced for comparison with YOLOV4-CSPDarknet53 in this study, which include YOLOV3-Darknet53, YOLOV3-MobileNetv3, SSD-MobileNetv3, and SSD-VGG16. A comprehensive comparison test was conducted to assess the classification and detection performance of these models for chicken parts. Finally, for the chicken part dataset, the mAP of the YOLOV4-CSPDarknet53 model was 98.86% on a single image with an inference speed of 22.2 ms, which was higher than the other four models of YOLOV3-Darknet53, YOLOV3-MobileNetv3, SSD-MobileNetv3, and SSD-VGG16 mAP by 3.27%, 3.78%, 6.91%, and 6.13%, respectively. The average detection time was reduced by 13, 1.9, 6.2, and 20.3 ms, respectively. In summary, the chicken part classification and detection method proposed in this study offers numerous benefits, including the ability to detect multiple chicken parts simultaneously, as well as delivering high levels of accuracy and speed. Furthermore, this approach effectively addresses the issue of accurately identifying individual chicken parts in the presence of occlusion, thereby reducing waste on the assembly line. PRACTICAL APPLICATION: The aim of this study is to offer visual technical assistance in minimizing wastage and resource depletion during the sorting and cutting of chicken parts in poultry production and processing facilities. Furthermore, considering the diverse demands and preferences regarding chicken parts, this research can facilitate product processing that caters specifically to consumer preferences.

The Spatial Features and Temporal Changes in the Gut Microbiota of a Healthy Chinese Population.

In this study, we aimed to understand the characteristics of the gut microbial composition in a healthy Chinese population and to evaluate if they differed across different regions. In addition, we aimed to understand the changes in the gut microbial composition over time. We collected 239 fecal samples from healthy Chinese adults living in four regions and performed a 1-year time cohort study in a small population in Beijing. The Chinese gut microbiota share 34 core bacterial genera and 39 core bacterial species, which exist in all collected samples. Several disease-related microorganisms (DRMs), virulence factors, and antibiotic resistance genes were found in one or more healthy Chinese samples. Differences in gut microbiota were observed in samples from different regions, locations, individuals, and time points. Compared to other factors, time was associated with a lower degree of change in the gut microbiota. Our findings revealed spatial and temporal changes in the gut microbiota of healthy Chinese individuals. Compared to fecal microbiomes of 152 samples in the publicly released the Human Microbiome Project (HMP) project from the United States, samples in this study have higher variability in the fecal microbiome, with higher richness, Shannon diversity indices, and Pielou evenness indexes, at both the genus and species levels. The microbiota data obtained in this study will provide a detailed basis for further understanding the composition of the gut microbiota in the healthy Chinese population. IMPORTANCE China accounts for approximately 1/5th of the world's total population. Differences in environment, ethnicity, and living habits could impart unique features to the structure of the gut microbiota of Chinese individuals. In 2016, we started to investigate healthy Chinese people and their gut microbiomes. Phase I results for 16S rRNA amplicons have been released. However, owing to the limitations of 16S rRNA amplicon sequencing, the gut microbiome of a healthy Chinese population could not be examined thoroughly at the species level, and the detailed changes in the gut microbiota over time need to be investigated. To address these knowledge gaps, we started a phase II study and investigated the basis for variations in the gut microbiome composition in a healthy Chinese population at the species level using shotgun metagenomics technology. In the phase II study, we also conducted a time scale analysis of fecal samples from healthy Chinese subjects, as a pioneered study, which quantitatively clarified the changes in the gut microbiota at both the spatial and temporal levels and elucidated the distribution pattern of DRMs in healthy Chinese individuals.

Time-scale analysis of the long-term variability of human gut microbiota characteristics in Chinese individuals.

Studying the dynamics and stability of the human gut microbiota over time is important for exploring their relationship with human health and developing treatment strategies for putative microbiome-related ailments. Here, we collected stool samples from seven healthy Chinese subjects at 1-month intervals between 2016 and 2020. Sequencing and bioinformatics analyses revealed that the bacteria in the collected fecal samples fluctuated over time, and the extent of these changes increased over time. Further, the average shared proportion value obtained using Sourcetracker2 was 63.5% for samples collected from the same individual in the preceding month, and over a 3-year period, this value decreased to 40.7%. Furthermore, the proportion of different bacteria in the gut microbiota of the different subjects fluctuated to varying degrees. Therefore, our results suggested that it is important to consider the effect of time on gut microbiota composition when it is used to evaluate health. Our study opens up a new field of microbiota research, considering not just the instantaneous microbiota, but also the change of the gut microbiota over time.

Cysteine string protein alpha accumulates with early pre-synaptic dysfunction in Alzheimer's disease.

In Alzheimer's disease, synapse loss causes memory and cognitive impairment. However, the mechanisms underlying synaptic degeneration in Alzheimer's disease are not well understood. In the hippocampus, alterations in the level of cysteine string protein alpha, a molecular co-chaperone at the pre-synaptic terminal, occur prior to reductions in synaptophysin, suggesting that it is a very sensitive marker of synapse degeneration in Alzheimer's. Here, we identify putative extracellular accumulations of cysteine string alpha protein, which are proximal to beta-amyloid deposits in post-mortem human Alzheimer's brain and in the brain of a transgenic mouse model of Alzheimer's disease. Cysteine string protein alpha, at least some of which is phosphorylated at serine 10, accumulates near the core of beta-amyloid deposits and does not co-localize with hyperphosphorylated tau, dystrophic neurites or glial cells. Using super-resolution microscopy and array tomography, cysteine string protein alpha was found to accumulate to a greater extent than other pre-synaptic proteins and at a comparatively great distance from the plaque core. This indicates that cysteine string protein alpha is most sensitive to being released from pre-synapses at low concentrations of beta-amyloid oligomers. Cysteine string protein alpha accumulations were also evident in other neurodegenerative diseases, including some fronto-temporal lobar dementias and Lewy body diseases, but only in the presence of amyloid plaques. Our findings are consistent with suggestions that pre-synapses are affected early in Alzheimer's disease, and they demonstrate that cysteine string protein alpha is a more sensitive marker for early pre-synaptic dysfunction than traditional synaptic markers. We suggest that cysteine string protein alpha should be used as a pathological marker for early synaptic disruption caused by beta-amyloid.

MDACP: A Pathogen Genome and Metagenome Analysis Cloud Platform.

Pathogenic microorganism analysis based on next-generation sequencing technology is an important tool for clinical diagnosis, public health surveillance, and outbreak investigation. However, scientific researchers without the relevant background lack the time, training, or infrastructure to use large data sets or install and use command line tools. Therefore, the bioinformatic team at the Chinese Center for Disease Control and Prevention developed the Microbial Data Analysis Cloud Platform (MDACP) as a safe, professional, and efficient pathogen genetic data analysis platform for rapid microbial data mining, such as for candidate pathogen detection, genome typing, and traceability. MDACP is a web service system based on the Docker platform and can be used for data analysis on various operating systems. The platform focuses on pathogen analysis and continuously develops new analysis processes according to the analysis needs of the users. This platform has a friendly user interface and is easy to operate, allowing users to submit data through data pages or graphical clients, flexibly control parameters according to data conditions, and analyze data in parallel with multiple tasks. Researchers can quickly carry out bioinformatic analyses without coding work, promote follow-up research and information mining of projects, and improve the utilization of big data in the field of disease control. MDACP enables research personnel to conduct data analysis and management and assists clinicians and disease control personnel with mining information, such as pathogen identification, classification, and traceability.

[A new quantitative 16S rRNA amplicon sequencing method].

In recent years, 16S rRNA amplicon sequencing technology has been widely used to study human gut microbiota and to detect unknown pathogens in clinical samples. However, its resolution to bacterial population can only reach the relative abundance of genus level, and different factors affect the final bacterial profile, such as sample concentrations, PCR cycle numbers and amplification primers. In order to solve these problems, we developed a quantitative 16S rRNA amplicon sequencing method by combining random tag and internal marker method. The new methods improved the accuracy of human gut microbiota, reduced the impact of experimental operation on the results, and improved the comparability between sequencing and other molecular biological methods.

A prospective study on peptide mapping of human fatigue saliva markers based on magnetic beads.

In order to explore convenient and stable fatigue markers, we studied various high-molecular-weight peptide fragments under fatigue state and non-fatigue state in the saliva using time of flight mass spectrometry. The saliva samples were collected from 10 healthy volunteers that were in the condition of fatigue and non-fatigue, respectively. Moreover, the time of flight mass spectrometry was conducted using two kinds of sample treatment methods, the magnetic beads enrichment (MB) and direct detection of stock solution. This was followed by modeling via the mass spectra of MB and supernatant (stock solution) directly collected after centrifugation. Both MB and direct sampling produced good spectrograms between 1,000 and 15,000 Da, while some peaks were lost in the enrichment. The spectrograms in the early and late period were different in each individual. Due to the limited sample size, 20 early and 20 late spectrograms were used for modeling analysis. Three different peptides were identified in the stock solution samples that can be detected in both fatigue and non-fatigue groups. The cross validity of MB model was 92.06%, while that of the stock solution model was 95.49%. The results showed that there were different peaks within the molecular weight of 2,000-15,000 Da, which provided a scientific basis for further realization of the convenient fatigue detection method based on the biosensor technique, with important theoretical and practical significance.

Gut microbiota community characteristics and disease-related microorganism pattern in a population of healthy Chinese people.

China's population accounts for about 1/5th of the world's total population. Owing to differences in environment, race, living habits, and other factors, the structure of the intestinal flora of Chinese individuals is expected to have unique features; however, this has not been thoroughly examined. Here, we collected faecal samples from healthy adults living in three cities of China and investigated their gut microbiome using metagenomics and bioinformatics technology. We found that 11 core bacterial genera were present in all of the Chinese faecal samples; moreover, several patient characteristics (age, region, body mass index, physical exercise, smoking habits, and alcoholic drink, and yogurt consumption) were found to have different effects on the gut microbiome of healthy Chinese people. We also examined the distribution patterns of disease-related microorganisms (DRMs), revealing which DRMs can potentially be used as markers for assessment of health risk. We also developed a program called "Guthealthy" for evaluating the health status associated with the microbiome and DRM pattern in the faecal samples. The microbiota data obtained in this study will provide a basis for a healthy gut microbiome composition in the Chinese population.

Gastric Juice-Based Real-Time PCR for Tailored Helicobacter Pylori Treatment: A Practical Approach.

A gastric juice-based real-time polymerase chain reaction (PCR) assay was established to identify Helicobacter pylori infection, clarithromycin susceptibility and human CYP2C19 genotypes and to guide the choice of proton pump inhibitor (PPI), clarithromycin and amoxicillin treatment for tailored H. pylori eradication therapy. From January 2013 to November 2014, 178 consecutive dyspeptic patients were enrolled for collection of gastric biopsy samples and gastric juice by endoscopy at the Peking University Third Hospital; 105 and 73 H. pylori-positive and -negative patients, respectively, were included in this study. H. pylori infection was defined as samples with both a strongly positive rapid urease test (RUT) and positive H. pylori histology. A series of primers and probes were distributed into four reactions for identifying the H. pylori cagH gene coupled with an internal control (Rnase P gene), A2142G and A2143G mutants of the H. pylori 23S rRNA gene, and single-nucleotide polymorphisms (SNPs) G681A of CYP2C19*2 and G636A of CYP2C19*3. The E-test and DNA sequencing were used to evaluate the H. pylori clarithromycin susceptibility phenotype and genotype. The SNPs CYP2C19*2 and CYP2C19*3 were also evaluated by nucleotide sequencing. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of this gastric juice-based real-time PCR assay were evaluated by comparing with the same measures obtained through gastric biopsy-based PCR and culture. The H. pylori diagnostic sensitivities of the culture, PCR, and gastric biopsy- and gastric juice-based real-time PCR assays were 90.48% (95/105), 92.38% (97/105), 97.14% (102/105) and 100% (105/105), respectively; the specificities of the above methods were all 100%. Higher false-negative rates were found among the gastric biopsy samples assessed by culture (10.48%, 11/105), PCR (7.62%, 8/105) and real-time PCR (2.86%, 3/105) than in gastric juice by real-time PCR. Regarding clarithromycin susceptibility, a concordance of 82.98% (78/94) and discordance of 17.02% (16/94) were observed among the different methods, discrepancies that mainly represent differences between the H. pylori clarithromycin susceptibility phenotype and genotype. Three coinfections of susceptible and resistant strains were detected, with resistant-to-susceptible ratios of 1.16, 3.44, and 8.26. The CYP2C19 genotyping results from gastric juice by real-time PCR were completely in accordance with those obtained from biopsy samples by conventional PCR. This gastric juice-based real-time PCR assay is a more accurate method for detecting H. pylori infection, clarithromycin susceptibility and CYP2C19 polymorphisms. The method may be employed to inform the choice of proton pump inhibitor (PPI), clarithromycin and amoxicillin treatment for tailored H. pylori eradication therapy.

Non-pylori Helicobacters (NHPHs) Induce Shifts in Gastric Microbiota in Helicobacter pylori-Infected Patients.

To explore the effects of gastric non-H. pylori Helicobacter species(NHPH) on the structure and potential function of gastric microbiota, we employed 16S rRNA gene sequencing on 164 gastric biopsy specimens from NHPH (H. suis, H. felis, H. salomonis) /H. pylori coinfection individuals, H. pylori monoinfection individuals and healthy controls. The results demonstrated that marked structural and functional variations between H. pylori mono- and coinfection samples (HPHS, HPHF, HPHM). The changes in bacterial structure induced by NHPH are mainly attributed to their ability of gastric acid secretion inhibition as well as bacterial chemotaxis. Both the HPHS and HPHF groups showed significant increases in phylotype richness and significant decreases in ß diversity, but this trend was not found in HPHM group. Regarding the top five phyla and top thirty-five genera, the HPHS and HPHF groups had similar variation trends in relative abundance. The increased relative abundance levels of the genera Vibrio, Pseudoalteromonas, Photobacterium, and Clostridium were associated with increases in predicted signal transduction/metabolic pathways among the three coinfection groups. The relative abundance levels of bacteria involved in the formation of N-nitroso compounds were significantly decreased in the HPHS and HPHF groups (e.g., Streptococcus, Neisseria, Haemophilus, Veillonella, Clostridium, etc.). The significantly decreased relative abundance levels of the phyla Firmicutes and Bacteroidetes in the HPHS and HPHF groups were associated with the observed increases in predicted lipid metabolism pathways. The results in this study implied that NHPH can arouse the variation of structure and function of gastric microbiota, which may pave the way to further research on the pathogenesis of gastric diseases.

The distribution of jhp0940, jhp0945, jhp0947, jhp0949 and jhp0951 genes of Helicobacter pylori in China.

BACKGROUND: The plasticity region of Helicobacter pylori (H. pylori) is a large chromosomal segment containing strain-specific genes. The prevalence of the plasticity region genes of the H. pylori strains in China remains unknown. The aim of this study was to examine the status of these genes and to assess the relationship between the genes and the diseases caused by H. pylori infection. METHODS: A total of 141 strains were isolated from patients with chronic active gastritis (CAG), peptic ulcer disease (PUD) and gastric carcinoma (GC). The prevalence of jhp0940, jhp0945, jhp0947, jhp0949 and jhp0951 was determined using PCR, and the results were analyzed using the chi-squared test. RESULTS: The prevalence rates of jhp0940, jhp0945, jhp0947, jhp0949 and jhp0951 in the H. pylori strains were 42.55, 51.06, 20.57, 56.03 and 63.12%, respectively. The prevalence rates of jhp0940 were similar in the isolates from the CAG, PUD and GC patients, and there was no association between the jhp0940 status and any of the diseases. In contrast, the prevalence rates of jhp0945, jhp0947, jhp0949 and jhp0951 were significantly higher in the PUD and GC isolates than in the CAG isolates (p < 0.01). A univariate analysis showed that jhp0945, jhp0947, jhp0949 and jhp0951 increased the risk of PUD, while only jhp0951 was significantly associated with PUD in the multivariate analysis (p = 0.0149). The jhp0945-positive isolates were significantly associated with an increased risk for GC (p = 0.0097). CONCLUSION: The plasticity region genes are widely distributed in Chinese patients, and a high prevalence of these genes occurs in more serious diseases. Therefore, jhp0951 status is an independent factor associated with the development of PUD, and jhp0945 may predict the future development of GC in patients with CAG and is considered to be the best candidate disease marker for H. pylori-related diseases.

Molecular typing of Chinese Streptococcus pyogenes isolates.

Streptococcus pyogenes causes human infections ranging from mild pharyngitis and impetigo to serious diseases including necrotizing fasciitis and streptococcal toxic shock syndrome. The objective of this study was to compare molecular emm typing and pulsed field gel electrophoresis (PFGE) with multiple-locus variable-number tandem-repeat analysis (MLVA) for genotyping of Chinese S. pyogenes isolates. Molecular emm typing and PFGE were performed using standard protocols. Seven variable number tandem repeat (VNTR) loci reported in a previous study were used to genotype 169 S. pyogenes geographically-diverse isolates from China isolated from a variety of disease syndromes. Multiple-locus variable-number tandem-repeat analysis provided greater discrimination between isolates when compared to emm typing and PFGE. Removal of a single VNTR locus (Spy2) reduced the sensitivity by only 0.7%, which suggests that Spy2 was not informative for the isolates screened. The results presented support the use of MLVA as a powerful epidemiological tool for genotyping S. pyogenes clinical isolates.

High natural variability bacteria identification and typing: Helicobacter pylori analysis based on peptide mass fingerprinting.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) coupled to the original Biotyper database has a poor ability to identify Helicobacter pylori. Furthermore, the existing typing methods for H. pylori have no obvious correlation with the virulence and pathogenicity of H. pylori in East Asia. In this study, MALDI-TOF MS Biotyper system (MBS) was used to identify and type H. pylori. In addition, label-free and bioinformatics techniques were used to reveal the protein components of different types of H. pylori. A total of 56 H. pylori isolates were added to the original reference database. For the 92 H. pylori strains validated, the identification efficiency at the species level was improved from 3 (3.2%) to 82 (89.1%) strains. A new ribotyping method for H. pylori based on peptide mass fingerprinting was developed. For P1 and P2 type H. pylori, respectively, 7 specific peaks at m/z 4320, 5202, 5246, 5268, 6066, 6941, and 7128 and 2 specific peaks at m/z 5246 and 6941 were found. Between P1 and P2 type strains, 62 proteins were significantly different. A total of 206 proteins for H. pylori identification and typing were identified, of which 110 were located on the inner cell membrane and 103 were located in the cytoplasm. The major classifications of these proteins were ribosomal proteins (15.5%) and enzymes (29.6%). MBS is suitable for the identification and typing of variable bacteria such as H. pylori, particularly if further super reference spectra are constructed. BIOLOGICAL SIGNIFICANCE: Helicobacter pylori (H. pylori) possesses very high genetic variability. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) coupled to the original Biotyper system (MBS) has a poor ability to identify H. pylori isolated from China. The identification capabilities of MBS for highly variable bacterial species remain to be established. On the other side, Scholars of East Asia and Western dispute the theory that there are obvious correlations between cagA and gastric cancer. The existing typing methods for H. pylori based on cagA gene have no obvious correlation with the virulence and pathogenicity of H. pylori in East Asia. In light of this phenomenon of Asian enigma, we suppose that there are other genes beyond cagA that are correlated with the virulence of H. pylori. Here, we improved the original database using numerous H. pylori isolated from different countries and raised the identification capability of MBS from 3.2% to 89.1%. A new ribotyping method for H. pylori based on peptide mass fingerprinting was developed. Furthermore, the protein components of H. pylori identification and typing were revealed. These findings thus provide a new way for H. pylori identification, typing and the research of pathogenic mechanism.