CiteSpace-based Document Information Visualization of Literature Published by Journal of Forensic Medicine from 1985 to 2018.

ABSTRACT: Objective To analyze a knowledge web of the literature published by Journal of Forensic Medicine from its founding in 1985 to 2018, describe the evolving process of forensic science research and explore the research hotspots and frontiers at present. Methods The literature that was published by Journal of Forensic Medicine from 1985 to 2018 was collected and analyzed in terms of elements, such as emerging research hotspots, high frequency keywords, authors, dispatching units, location of institution and funding, by CiteSpace5.3 information visualization analysis software. Results All disciplines of forensic medicine were continually developing and maturing, and the publication volume of the literature on forensic pathology had the highest weight; in research hotspots, the two categories, research and identification each had their own emphasis; as the main source of contributions to the journal, research institutes accounted for 38.99% of the total number of publications; Shanghai ranked first among all regions with 1 046 articles published. The number of funded articles was generally on the rise, with the number of funded articles published largest in 2015. Conclusion As an authoritative academic journal in the field of forensic science in China, Journal of Forensic Medicine carries the development of forensic science and witnesses the institutional reform of universities and colleges, and offers a wide range of communication and cooperation in terms of technicality and application. Many scholars and scientific research institutions have gained progress continually in various research directions in the form of teamwork; and emerging research hotspots will continue to play a huge role in future practical applications.

Gonadal morphogenesis and sex differentiation in cultured Ussuri catfish Tachysurus ussuriensis.

The objective of this study was to investigate the optimal developmental time to perform sex reversal in Ussuri catfish Tachysurus ussuriensis, to develop monosex breeding in aquaculture. Systematic observations of gonadal sex differentiation of P. ussiriensis were conducted. The genital ridge formed at 9 days post fertilization (dpf) and germ cells begin to proliferate at 17 dpf. The ovarian cavity began forming on 21 dpf and completed by 25 dpf while presumptive testis remained quiescent. The primary oocytes were at the chromatin nucleolus stage by 30 dpf, the peri-nucleolus stage by 44 dpf and the cortical alveoli stage by 64 dpf. The germinal vesicle migrated towards the animal pole (polarization) at 120 dpf. In presumptive testis, germ cells entered into mitosis and blood vessels appeared in the proximal gonad on 30 dpf. The efferent duct anlage appeared on 36 dpf and formation of seminal lobules with spermatogonia and lobules interstitium occurred at 120 dpf. Therefore, gonadal sex differentiation occurred earlier in females than in males, with the histological differentiation preceding cytologic differentiation in T. ussuriensis. This indicates that undifferentiated gonads directly differentiate into ovary or testis between 17 and 21 dpf and artificial induction of sexual reversal by oral steroid administration must be conducted before 17 dpf.

Plasmacytoid dendritic cells and type 1 interferon promote peripheral expansion of forkhead box protein 3(+) regulatory T cells specific for the ubiquitous RNA-binding nuclear antigen La/Sjögren's syndrome (SS)-B.

RNA-binding nuclear antigens are a major class of self-antigen to which immune tolerance is lost in rheumatic diseases. Serological tolerance to one such antigen, La/Sjögren's syndrome (SS)-B (La), is controlled by CD4(+) T cells. This study investigated peripheral tolerance to human La (hLa) by tracking the fate of hLa-specific CD4(+) T cells expressing the transgenic (Tg) 3B5.8 T cell receptor (TCR) after adoptive transfer into lymphocyte-replete recipient mice expressing hLa as a neo-self-antigen. After initial antigen-specific cell division, hLa-specific donor CD4(+) T cells expressed forkhead box protein 3 (FoxP3). Donor cells retrieved from hLa Tg recipients displayed impaired proliferation and secreted interleukin (IL)-10 in vitro in response to antigenic stimulation. Transfer of highly purified FoxP3-negative donor cells demonstrated that accumulation of hLa-specific regulatory T cells (Treg ) was due primarily to expansion of small numbers of donor Treg . Depletion of recipient plasmacytoid dendritic cells (pDC), but not B cells, severely hampered the accumulation of FoxP3(+) donor Treg in hLa Tg recipients. Recipient pDC expressed tolerogenic markers and higher levels of co-stimulatory and co-inhibitory molecules than B cells. Adoptive transfer of hLa peptide-loaded pDC into mice lacking expression of hLa recapitulated the accumulation of hLa-specific Treg . Blockade of the type 1 interferon (IFN) receptor in hLa Tg recipients of hLa-specific T cells impaired FoxP3(+) donor T cell accumulation. Therefore, peripheral expansion of Treg specific for an RNA-binding nuclear antigen is mediated by antigen-presenting pDC in a type 1 IFN-dependent manner. These results reveal a regulatory function of pDC in controlling autoreactivity to RNA-binding nuclear antigens.

Anti-idiotypic antibodies prevent the serologic detection of antiribosomal P autoantibodies in healthy adults.

A subset of SLE patients has serologically detectable autoantibodies to the ribosomal P proteins (anti-P). We reported the discovery of covert anti-P antibodies and their masking IgG-inhibitory antibodies in the sera of healthy adults. The aim of this study was to determine if these IgG-inhibitory antibodies are anti-idiotypic antibodies (anti-Ids). IgG and IgG-depleted fractions of plasma from two healthy adults were assayed for inhibition of anti-P F(ab')2 binding to the ribosomal P proteins in immunoblot. Anti-P antibody activity was completely inhibited by plasma IgG, whereas there was no inhibition by IgG-depleted plasma. IgG-inhibitory antibodies recognized a cross-reactive epitope among anti-P from different SLE patients. Plasma IgG from one healthy adult was depleted of pepsin agglutinators and generic anti-F(ab')2 antibodies by adsorption with an affinity column prepared with normal IgG F(ab')2. Unretained IgG bound exclusively to anti-P F(ab')2 in ELISA. Using four affinity columns, we isolated IgG anti-Ids to anti-P antibodies from four healthy adults. These purified anti-Ids bound to anti-P F(ab')2 from a healthy adult and SLE patients. They did not bind to F(ab')2 fragments prepared from normal IgG or anti-dsDNA. Ribosomal antigens blocked this anti-Id-Id interaction. Purified anti-Ids inhibited the binding of anti-P F(ab')2 from patients to ribosomal P proteins. SLE patients without overt anti-P antibodies also possessed IgG anti-Ids to anti-P antibodies. We conclude that IgG-inhibitory antibodies are anti-Ids to anti-P antibodies, and are directed to public idiotopes on anti-P antibodies. These anti-Ids may be part of an Id network that regulates anti-P antibody expression, and perhaps pathogenicity.

Robust thumb flexor tendon repairs with a six-strand M-Tang method, pulley venting, and early active motion.

We present the outcomes of flexor pollicis longus tendon repairs in 34 thumbs using a six-strand M-Tang repair with venting of one or two pulleys according to site of tendon laceration. The A2 pulley was vented in all three thumbs with zone 1 injury. In 31 thumbs with zone 2 injuries, the oblique pulley was vented partially or entirely. Twenty-two thumbs had both the A1 and oblique pulleys vented. Six to 46 months post-surgery, 14 thumbs with zone 2 injuries were rated excellent, 13 good, three fair and one failure according to Tang criteria. No tendon ruptures or bowstringing occurred. Fourteen of 34 thumbs had deficits in interphalangeal joint extension averaging 13°. We conclude that venting of one or two pulleys may ensure recovery of thumb function without risking tendon bowstringing and that early active thumb motion is safe with a robust tendon repair. LEVEL OF EVIDENCE: IV.

The effect of curcumin on bladder tumor in rat model.

OBJECTIVE: Bladder cancer is the most commonly malignant tumor in the urogenital tract, only next to prostate cancer with a higher incidence in China. Curcumin is the major component of curcuma longa and has multiple biological effects including anti-tumor. This study aimed to investigate the effect of curcumin on bladder cancer. MATERIALS AND METHODS: SPF-grade Wistar rats were used for establishing bladder cancer model through injection of N-methyl-N-nitrosourea (MNU). Rats were then randomly divided into experimental, model and control group. 160 µmol/L curcumin were applied in the experimental group while model group received an equal volume of saline. General condition, morphology changes and cell cycle of bladder cancer cells were examined. Meanwhile, apoptotic proteins including Bcl-2, Bax and surviving were also measured by Western blot. RESULTS: Model rats displayed fever, hematuria, decreased food and water intake, dispersed fur, lower body mass and decreased activity. Under microscopy, the bladder wall was thickened with the cauliflower-like lesion, in which significant necrotic and hemorrhagic lesions were found. Experimental group rats improved general condition without decrease of body mass. The only minor lesion was found without significant necrosis or hemorrhage without invasion into the muscular layer. The number of G1 phase cells was increased while S phase cell number was decreased after drug intervention, suggesting suppression of G1/S transition (p < 0.05). In curcumin-treated rats, the expression of Bcl-2 and Survivin were significantly decreased while Bax protein expression was significantly elevated (p < 0.05). CONCLUSIONS: Curcumin can inhibit the growth and invasion of rat bladder cancer cells, possibly through the arresting of G1/S transition and subsequently increased apoptosis.

[Effects of IL10-592 locus of AA genotype on the incidence of aGVHD and survival after HLA-matched unrelated allogeneic hematopoietic stem cell transplantation].

OBJECTIVE: To explore the impact of IL10-592 (rs1800872) single nucleic acid polymorphism (SNP) on the prognosis of HLA matched unrelated hematopoietic stem cell transplantation (HSCT). METHODS: The polymorphism of IL10-592 in 104 recipient-donor pairs and 100 healthy volunteers was analyzed with sequence based typing (SBT). RESULTS: When the genotype of IL10-592 in donors and recipients matched, AA/AA genotype had higher incidence of Ⅲ-Ⅳ aGVHD than AC/AC or CC/CC genotype (47.1%, 3.7%, 0, P=0.002). When the genotype of IL10-592 in donors and recipients mismatched, recipients with AC genotype or donors with AA genotype, there was significant different incidence of Ⅲ-ⅣaGVHD among donors or recipients with different genotype (P=0.046, P=0.041). The recipients with AA genotype had higher incidence of Ⅲ-Ⅳ aGVHD than AC or CC genotype (27.8% vs 10.2%, 11.1%; P=0.072), and higher incidence of intestinal aGVHD (22.2% vs 5.1%,11.1%; P=0.040) , lower incidence of 2-year overall survival (OS: 48.2% vs 75.1%, 85.7%; P=0.002), lower incidence of 2 year disease free survival (DFS: 48.5% vs 66.3%, 76.2%; P=0.045). Patients had higher incidence of Ⅲ-Ⅳ aGVHD with donors of AA genotype than with donors of AC or CC genotype (26.5% vs 8.9%, 0; P= 0.024), and higher incidence of intestinal aGVHD (20.4% vs 4.4%, 0; P=0.026). In multivariate analysis, the genotype of IL10-592AA in recipients and donors had increased risk of Ⅲ-Ⅳ aGVHD (OR=3.3, P= 0.049; OR=3.9, P=0.043). There were no statistical differences on the incidence of cGVHD and relapse. CONCLUSION: In HLA-10/10 matched unrelated HSCT, the presence of IL10-592 AA genotype in recipients and/or donors is an adverse factor for Ⅲ-ⅣaGVHD, worse OS and 2-year DFS.

[The impact of HLA haplotype and alleles mismatches of donor-recipient pairs on outcome of haploidentical hematopoietic stem cell transplantation with a third part cord blood unit].

OBJECTIVE: To analyze allele mismatches of HLA- A, - B, - C, - DRB1, - DQB1 and haplotype mismatch of donor- recipient pairs on the outcome of haploidentical transplantation combined with a third part cord blood unit. METHODS: 230 pairs of donor-recipient were performed HLA-A, B, C, DRB1, DQB1 typing using SBT and SSOP methods from January 2012 to December 2014. RESULTS: Pairs were divided into HLA- 5/10、6/10、7/10 and ≥8/10 groups according to HLA- A, B, C and DRB1 highresolution typing and matched degrees, the 3-year probability of overall survival (OS) for each group were 48.7%, 59.3%, 71.1%, 38.3% (P=0.068) respectively. HLA-6/10 matched group associated with significant favorable effect on OS compared with HLA- 5/10 matched one (P=0.041).When the HLA class I antigen matched on the recipient and donor, improved OS and event free survival (EFS) in HLA- 6/10 matched group than in HLA-5/10 matched one (P=0.017,P=0.088), especially in single HLA-A loci allele matched one (P=0.013,P=0.013), were observed. As to the third part cord blood unit, sharing the same haplotype with the recipient-donor pairs produced better platelet recovery than the misfit one (95.3%vs 86.2%,P= 0.007), similar result was found in terms of neutrophil recovery (98.8%vs 96.1% ,P=0.022). CONCLUSIONS: HLA locus mismatch and haplotype mismatch of the donor and recipient should be useful for selection of the most optimum donor. Co- infused of an unrelated cord blood unit sharing the same haplotype with the recipient-donor pairs could improve hematopoietic recovery.

Enzyme-linked immunosorbent assay for detection of immunoglobulin G antibody against Helicobacter pylori.

Eight strains of sonicated Helicobacter pylori (H. pylori) were pooled as group antigen to detect the serum IgG antibody to H. pylori in patients with various gastroduodenal diseases and subjects at health check-up. The enzyme-linked immunosorbent assay method used had a sensitivity of 94%, a specificity of 87%, and a positive predictive value of 96%. The prevalence of serum IgG antibody to H. pylori was 19.0% in patients with Type A gastritis, 76.0% in patients with Type B gastritis, 91.5% in duodenal ulcer, and 92.9% in gastric ulcer. Moreover, the prevalence of serum IgG antibody to H. Pylori in newborns was 53% (probably acquired from mother), in children below 10 years old was 25%. The detection rate gradually rose to 70% with the increase of age, but decreased to 40% in those over 70 years of age.

Proteomic analysis of human bone marrow mesenchymal stem cells transduced with human telomerase reverse transcriptase gene during proliferation.

OBJECTIVES: Previous studies have reported immortalization and tumorigenicity of human mesenchymal stem cells (hMSCs) transduced with exogenous human telomerase reverse transcriptase (hTERT). We also have established a line of hMSCs transduced with hTERT (hTERT-hMSCs) and we have cultured these cells for 290 population doublings (PDs) during which they demonstrated a large proliferation potential but with no tumorigenicity. The aim of this study was to investigate the protein expression profile of hTERT-hMSCs with two-dimensional gel electrophoresis and peptide mass fingerprinting by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, to be able to analyse the effects of exogenous hTERT on protein expression in hMSCs. MATERIALS AND METHODS: We generated proteome maps of primary hMSCs and hTERT-hMSCs at PD 95 and PD 275. RESULTS: A total of 1543 +/- 145 protein spots in gels of primary MSCs at PD 12, 1611 +/- 186 protein spots in gels of hTERT-hMSCs at PD 95 and 1451 +/- 126 protein spots in gels of hTERT-hMSCs at 275 PD were detected. One hundred of these were successfully identified, including 20 which were differentially expressed. CONCLUSIONS: The results suggest that sustaining levels of prohibitin and p53 expression along with differential expression of proteins in hTERT-hMSCs provide an insight into lack of transforming activity of hTERT-hMSCs during cell proliferation.

Neo-epitopes are required for immunogenicity of the La/SS-B nuclear antigen in the context of late apoptotic cells.

Mechanisms responsible for the induction of anti-nuclear autoantibodies (ANA) following exposure of the immune system to an excess of apoptotic cells are incompletely understood. In this study, the immunogenicity of late apoptotic cells expressing heterologous or syngeneic forms of La/SS-B was investigated following subcutaneous administration to A/J mice, a non-autoimmune strain in which the La antigenic system is well understood. Immunization of A/J mice with late apoptotic thymocytes taken from mice transgenic (Tg) for the human La (hLa) nuclear antigen resulted in the production of IgG ANA specific for human and mouse forms of La in the absence of foreign adjuvants. Preparations of phenotypically healthy cells expressing heterologous hLa were also immunogenic. However, hLa Tg late apoptotic cells accelerated and enhanced the apparent heterologous healthy cell-induced anti-La humoral response, while non-Tg late apoptotic cells did not. Subcutaneous administration of late apoptotic cells was insufficient to break existing tolerance to the hLa antigen in hLa Tg mice or to the endogenous mouse La (mLa) antigen in A/J mice immunized with syngeneic thymocytes, indicating a requirement for the presence of heterologous epitopes for anti-La ANA production. Lymph node dendritic cells (DC) but not B cells isolated from non-Tg mice injected with hLa Tg late apoptotic cells presented immunodominant T helper cell epitopes of hLa. These studies support a model in which the generation of neo-T cell epitopes is required for loss of tolerance to nuclear proteins after exposure of the healthy immune system to an excess of cells in late stages of apoptosis.

Recombinant ribosomal P2 protein can unmask anti-ribosomal P autoantibodies from healthy adults.

Autoantibodies to ribosomal P proteins (anti-P) are detected almost exclusively in the serum samples from patients with systemic lupus erythematosus when conventional enzyme-linked immunosorbent assay and immunoblotting techniques are used. Anti-P are not detected in serum samples from healthy adults by these techniques. By treating serum from healthy adults with ribosome-coated beads, we unexpectedly unmasked anti-P in virtually all individuals. This unmasking of anti-P occurs by the displacement of an antibody inhibitor from anti-P. We wanted to determine whether anti-P from healthy adults could also be unmasked by treatment of their serum or plasma with isolated ribosomal P proteins. Recombinant human ribosomal P2 protein was produced in bacteria as a TrpE fusion protein, resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, blotted onto nitrocellulose membranes, and isolated as strips of membranes corresponding to the size of the P2 fusion protein. Serum or plasma from six healthy adults and three patients with systemic lupus erythematosus were incubated with these strips overnight rather than for 2 hours, as is done in conventional immunoblots. Acid eluates were obtained from the strips and analyzed for antibody activity by immunoblot. Eluates from healthy adults and patients contained antibodies reactive with recombinant ribosomal P2 protein. They also reacted with the three ribosomal P proteins in purified rabbit ribosomes. Their anti-P activity was completely inhibited by a peptide corresponding to the immunodominant linear epitope of the ribosomal P proteins. The antibodies in the eluate were immunoglobulin G. We conclude that anti-P autoantibodies from healthy adults can be unmasked by affinity purification on denatured, recombinant ribosomal P proteins and that antigen excess is sufficient for inhibitor displacement.

A murine monoclonal anti-idiotype to anti-ribosomal P antibodies: production, characterization, and use in systemic lupus erythematosus.

Overt anti-ribosomal P (anti-P) autoantibodies are restricted to a subset of systemic lupus erythematosus (SLE) patients, and are potentially pathogenic. Covert anti-P are detected in all other individuals. An idiotype (Id) network is nonoperational in those with overt anti-P, whereas it is functional in all others. The aim of this study was to produce a murine monoclonal (mAb) anti-Id to characterize the anti-P Id network in SLE. BALB/c mice were immunized with F(ab')(2) fragments of IgG anti-P from a patient with a broadly cross-reactive Id. One mAb was chosen (mAb41) that reacted preferentially to the immunogen. This IgG1 mAb bound comparably in ELISAs to affinity-purified anti-P from 11 SLE patients with overt anti-P. This binding was partially inhibited with ribosomal P antigen. In contrast, it did not bind to affinity-purified control autoantibodies, nor to normal human IgG. mAb41 inhibited anti-P binding to ribosomal P antigen in immunoassays and on Jurkat cells. No change was detected in patients' anti-P antibodies over time when mAb41 was used in Id-specific ELISAs. We conclude that mAb41 is an anti-Id that recognizes a public idiotope within the antigen-combining site of anti-P antibodies. Thus, it is analogous to its human counterparts, and potentially, would modulate the pathogenicity of anti-P autoantibodies in vivo.

T-helper cell tolerance to ubiquitous nuclear antigens.

Systemic autoimmune diseases are characterized by the development of antinuclear autoantibodies. In order to understand the immunologic events leading to the development of such antibodies, knowledge of mechanisms of immune tolerance to nuclear antigens is required. By utilizing adoptive T-cell transfer strategies with transgenic mouse models expressing nuclear neo-self antigens, T-cell tolerance to the lupus-related nuclear antigens human La and nRNP A has been demonstrated. These findings also indicate the existence in normal animals of autoreactive B cells continuously presenting nuclear antigen, suggesting that nuclear antigens are not sequestered from the immune system. Investigations of CD4+ T-cell tolerance to non-nuclear antigens have revealed a number of mechanisms that protect the host from autoreactivity, including autoreactive T-cell deletion, regulatory T-cell development and anergy induction. Recent studies using T-cell receptor and neo-self nuclear antigen transgenic mice are revealing the importance of such mechanisms in maintaining tolerance to nuclear antigens. Mechanisms of tolerogenic antigen presentation, identification of tolerogenic antigen source(s) and the pathways leading to loss of tolerance to nuclear antigens in systemic autoimmune disease states are currently being sought.

Relation between vacA subtypes, cytotoxin activity, and disease in Helicobacter pylori-infected patients from The Netherlands.

OBJECTIVE: The vacuolating cytotoxin of Helicobacter pylori (H. pylori) is encoded by vacA, of which allelic variation has been described. In the U.S., H. pylori strains with the signal sequence allele s1a are associated with enhanced gastric inflammation and with peptic ulcer disease (PUD). The m1 middle region allele is linked with more severe gastric epithelial damage. However, the distribution of H. pylori genotypes and the association with disease may be different in other geographical areas. The aim of this study was to establish whether vacA types among H. pylori isolates from Dutch patients are associated with disease. METHODS: The cytotoxin activity of the H. pylori isolates from 34 PUD patients and 46 patients with functional dyspepsia (FD) was assessed by an in vitro assay using HeLa cells as indicator cells. The vacA types and cagA status of the isolates were assessed by polymerase chain reaction (PCR). RESULTS: vacA s1-type H. pylori displayed cytotoxin activity more frequently than s2 vacA-type H. pylori (p = 0.003). This difference was not significant when only cagA+ H. pylori were considered. H. pylori isolates with the m1 vacA type exhibited a higher cytotoxin activity, independent of cagA (p = 0.006). Ninety-four percent (32/34) of the PUD patients and 74% (34/46) of the FD patients were infected with s1 vacA-type H. pylori (p = 0.04). When only cagA+ H. pylori were considered, s1 vacA type was not associated with disease. In addition, neither the s1a nor s1b subtypes correlated with disease. CONCLUSIONS: An association between vacA subtypes and disease could not be established in this patient population, due to the strong linkage between vacA s1 type and cagA.

Equally high prevalences of infection with cagA-positive Helicobacter pylori in Chinese patients with peptic ulcer disease and those with chronic gastritis-associated dyspepsia.

Approximately 60% of Helicobacter pylori isolates in the Western world possess the cytotoxin-associated gene A (cagA). cagA-positive H. pylori is found to be associated with peptic ulcer disease (PUD) and gastric adenocarcinoma. To investigate the cagA status of H. pylori isolates from Chinese patients with PUD and chronic gastritis (CG), H. pylori populations from 83 patients, 48 with PUD and 35 with CG, were assessed by two different cagA-specific PCRs, Southern blotting, and colony hybridization. The combined results from PCR, Southern blotting, and colony hybridization indicate a prevalence of cagA-positive H. pylori isolates of 98% (47 of 48) among Chinese PUD patients and 100% (35 of 35) among Chinese CG patients. Amplification with primer sets 1 and 2 yielded 52 and 95% of the 82 cagA-positive Chinese H. pylori, respectively. In contrast, the sensitivity of cagA-specific PCR for cagA-positive H. pylori isolates from Dutch patients with primer set 1 was 92% (112 of 122) and that with primer set 2 was 91% (50 of 55). The prevalence of cagA-positive H. pylori populations in Chinese patients with PUD and CG is almost universally high. Therefore, cagA cannot be used as a marker for the presence of PUD in Chinese patients. Our data further suggest that allelic variation in cagA may exist and that distinct H. pylori genotypes may circulate in China and Western Europe.

cagA-positive Helicobacter pylori populations in China and The Netherlands are distinct.

The aim of this research was to study whether and to what extent Chinese cagA-positive Helicobacter pylori isolates differ from those in The Netherlands. Analysis of random amplified polymorphic DNA (RAPD)-PCR-assessed DNA fingerprints of chromosomal DNA of 24 cagA-positive H. pylori isolates from Dutch (n = 12) and Chinese (n = 10) patients yielded the absence of clustering. Based on comparison of the sequence of a 243-nucleotide part of cagA, the Dutch (group I) and Chinese (group II) H. pylori isolates formed two separate branches with high confidence limits in the phylogenetic tree. These two clusters were not observed when the sequence of a 240-bp part of glmM was used in the comparison. The number of nonsynonymous substitutions was much higher in cagA than in glmM, indicating positive selection. The average levels of divergence of cagA at the nucleotide and protein levels between group I and II isolates were found to be high, 13.3 and 17.9%, respectively. Possibly, the pathogenicity island (PAI) that has been integrated into the chromosome of the ancestor of H. pylori now circulating in China contained a different cagA than the PAI that has been integrated into the chromosome of the ancestor of H. pylori now circulating in The Netherlands. We conclude that in China and The Netherlands, two distinct cagA-positive H. pylori populations are circulating.