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Polo-like kinase 1 (PLK1) is overexpressed in primary colorectal cancers.

PLK (polo-like kinase), the human counterpart of polo in Drosophila melanogaster and of CDC5 in Saccharomyces cerevisiae, belongs to a family of serine/threonine kinases. It is intimately involved in spindle formation and chromosome segregation during mitosis. The purpose of this study was to determine whether PLK1 is overexpressed in primary colorectal cancer specimens as compared with normal colon mucosa and to assess its relation to other kinases as a potential new tumor marker. In the present study, immunohistochemical analyses were performed of PLK1 expression in 78 primary colorectal cancers as well as 15 normal colorectal specimens. Furthermore, we examined the relationship between other kinases, Aurora-A and Aurora-C, and PLK1 expression. In normal colon mucosa, some crypt cells showed weakly positive staining for PLK1 in 13 out of 15 cases, the remaining cases being negative. Elevated expression of PLK1 was observed in 57 (73.1%) of the colorectal cancers, statistically significant associations being evident with pT (primary tumor invasion) (P=0.0006, Mann-Whitney U test), pN (regional lymph nodes) (P=0.008, chi2 test) and the Dukes' classification (P=0.0005, Mann-Whitney U test). Mean proliferating cell nuclear antigen-labeling index was 52.3%, with a range of 24.1% to 77.3%. Values for lesions with high and low PLK1 expression were 54.7+/-10.3% (mean+/-SD) and 45.9+/-11.9% (P=0.002, Student's t test). PLK1 was significantly associated with Aurora-A, but PLK1 staining was more diffuse and extensive than for Aurora-A or Aurora-C. Interestingly, PLK1 overexpression was significantly associated with p53 accumulation in colorectal cancers. Our results suggest overexpression of PLK1 might be of pathogenic, prognostic and proliferative importance, so that this kinase might have potential as a new tumor marker for colorectal cancers.

Adenocarcinoma↗

Rapid diagnosis of avian influenza (AI) and assessment of pathogenicity of avian H5 and H7 subtypes by molecular methods.

Avian influenza (AI) is a highly contagious viral disease of poultry that is found worldwide. There are two forms of AI: a mild form called low pathogenicity avian influenza (LPAI), and a severe form called highly pathogenic avian influenza (HPAI). HPAI is associated with the H5 and H7 subtypes of AI virus (AIV) and is subject to Federal control and International reporting. A real-time reverse transcription-polymerase chain reaction (rRT-PCR) assay has been developed and validated that can help in the early detection of AI outbreaks. The rRT-PCR assay can also be used to identify infections caused by H5 and H7 subtypes of AIV New isolates of AIV must be characterized as LPAI or HPAI for reporting and control purposes. The criteria for classification of an AI virus as HPAI are defined by the World Organization for Animal Health (OIE); the definition includes a virulence and a molecular criterion. The virulence requirement for HPAI is defined as an AIV killing 75% or more of eight inoculated chickens within 10 days. The molecular criterion is the presence of multiple dibasic amino acids at the proteolytic cleavage site of the haemagglutinin (H) protein. All HPAI viruses isolated before 2002 fulfilled both the virulence and molecular criteria. Consequently, nucleotide sequencing of the H gene and deduction of the amino acid motif at the H cleavage site has been successfully used to assess the virulence of H5 and H7 AIVs rapidly. Since 2002, however, there have been three outbreaks of HPAI where the viruses responsible for the outbreaks have either fulfilled the virulence criterion or the molecular criterion, but not both.

Amino Acid Sequence↗

[Vascular abnormalities].

Formerly, if one would decide to get some interest in vascular anomalies, one would be abashed by the multiplicity of classifications, the empiricism of treatments but mainly by the poverty of fundamental research due to the lack of animal models. Today progress in molecular Biology and imaging allow to establish a coherent classification of vascular anomalies. Thanks to the development of quantum physics, pulsed dye laser is very efficacious in the treatment of capillary malformations (port-wine stains). Plastic surgery and reconstructive microsurgery allow wide resection of venous and arteriovenous malformations in collaboration with interventional radiology. Finally, recent discoveries in molecular genetic will allow the creation of transgenic mice with vascular anomalies phenotypically similar to human malformations. So, in the near future, we will be able to test experimentally new therapeutic approaches but especially set up research protocols directed to a better understanding of the pathogenic mechanisms of these vascular anomalies.

Animals↗

[Probe into internal relation between classification of the differentiation-syndrome in traditional Chinese medicine and serum copper and zinc in lung cancer].

This article probed into the internal relations and significance between differentiation of syndrome of traditional Chinese medicine (TCM) and trace elements--copper and zinc in lung cancer patients. The serum copper and zinc of 95 patients with lung cancer and 82 healthy persons were measured. According to differentiation of syndrome of TCM types of 95 lung cancer patients were divided, and the relations with their levels of serum copper, zinc and the ratio of copper/zinc compared respectively. The authors found that there were some inner links among the differentiation syndromes and levels of serum copper, zinc and its ratio. The result showed that the level of Cu/Zn ratio could reflect increase and decrease of body resistance and pathogenic factors the level of Cu/Zn ratio was more significant than that of copper and zinc. The authors suggested that the ratio of Cu/Zn could be used as the criteria of differentiation of syndrome of TCM. It is clinically significant to combine the level of copper, zinc and its ratio with differentiation of syndrome of TCM to evaluate the severity and prognosis of the patients and to direct the treatment of them with TCM.

Adenocarcinoma↗

Bacterial plasmids--an overview.

Resistance to antimicrobial substances and various factors contributing to pathogenicity are only some of the bacterial traits that can be determined by plasmid genes. Not all plasmids are involved with resistance, nor is all resistance to antimicrobials plasmid-mediated. Nevertheless, the impact of R-plasmids on the choice of antimicrobial therapy is substantial, especially in hospital and when dealing with certain community pathogens, such as Haemophilus influenzae. The discovery of transposition elements has helped us to understand the rapidity with which certain resistance determinants develop and become disseminated among diverse microbial species. Similarly, not all determinants of microbial pathogenicity are plasmid encoded and the genes for many toxins, cellular attachment pili, iron sequestration systems, etc. are found on the bacterial chromosomes. In Yersinia, pathogenicity is even more complex in that plasmid determinants play an essential role, but only in concert with chromosomal genes. The particularly significant features of such pathogenicity-determined plasmid-mediated genes is not that they are found on extrachromosomal elements, but that they may possess greater genetic mobility. Another consequence of the presence of virulence genes on bacterial plasmids is that it makes them easier to study by the newer techniques of microbial genetics and molecular biology.

Anti-Bacterial Agents↗

Comprehensive classification of symptoms and signs reported among 218 patients with acute HIV-1 infection.

Acute HIV-1 illness presents a wide range of clinical manifestations. We assessed a classification and data reduction of clinical features among 218 patients with acute HIV-1 infection enrolled in four prospective seroincidence studies. Factor analysis allows the definition of eight factors based on groups of symptoms and signs: gastrointestinal transit disturbances, weight loss/abdominal pain, lymphadenopathy, myalgia/arthralgia, neurologic features, constitutional and mucocutaneous features, oral candidiasis, and anorexia/pharyngitis. These groups reflected the main target systems involved at the time of acute HIV-1 disease. Grouping of symptoms and signs based on groups of patients is potentially more informative than observations made on individual patients. It might facilitate diagnosis, suggest pathogenic mechanisms and reduce the number of variables for characterizing acute HIV-1 illness.

Acute Disease↗

Spondyloarthropathy is underestimated in inflammatory bowel disease: prevalence and HLA association.

OBJECTIVE: To determine the overall prevalence of spondyloarthropathy (SpA) among patients with inflammatory bowel disease (IBD) [Crohn's disease (CD) and ulcerative colitis (UC)]. METHODS: One hundred three consecutive patients with IBD from a gastroenterology unit were questioned and examined for SpA symptoms. Patients previously diagnosed with SpA were excluded. All patients were questioned and examined for SpA symptoms such as inflammatory back pain, joint swelling, enthesitis, and psoriasis or a specific family history. Radiographs were taken of all sacroiliac joints. HLA loci A, B, C, and DR were determined in all patients. RESULTS: Thirty-nine percent of the patients with IBD had clinical articular manifestations: 30% had inflammatory back pain, 10% had synovitis, and 7% had a peripheral enthesopathy. The majority (90%) of patients with rheumatic complaints fulfilled the classification criteria for SpA and 10% fulfilled the criteria for ankylosing spondylitis. Asymptomatic sacroiliitis was found in an additional 18% of the patients. Moreover, sacroiliitis, symptomatic or asymptomatic, was related to the disease duration. HLA-B27 conferred an additional risk for inflammatory low back pain in patients with IBD. CONCLUSION: Articular involvement in IBD can be classified as SpA. The appearance of SpA occurs irrespective of the extent of the bowel disease. Moreover, asymptomatic sacroiliac involvement is a common manifestation in IBD and it is related to disease duration, suggesting evidence for a related pathogenic mechanism.

Aged↗

[Evaluation of trends in ue of antibiotics and resistance in selected bacterial pathogens].

BACKGROUND: Evaluation of antibiotic consumption and the surveillance of bacterial pathogens resistance appears to be an indispensable part of antibiotic policy. The aim of this study was to evaluate dynamics of antibiotics consumption and resistance development in selected bacterial strains (Pseudomonas aueruginosa, Staphylococcus aureus and Staphylococcus epidermidis) in the environment of Faculty Hospital. METHODS AND RESULTS: The analysis was focused on the development of antibiotic consumption and monitoring of the development of bacterial resistance. Samples were taken in internal and surgical departments of General Faculty Hospital in Prague in 1992, 1995, and 1999. The evaluation of antibiotic consumption was based on Anatomical Therapeutic and Chemical classification system and the Defined Daily Doses (DDD). The consumption in the follow up period almost doubled in surgical department while in internal medicine department was decreased by almost one third. In both department, namely in the surgical one significant increase of bacterial resistance of tested antibiotics was noted. CONCLUSION: The important part of the local antibiotic policy is the monitoring of development of bacterial resistance and consumption of antibiotics.

Anti-Bacterial Agents↗

Hepatitis C virus genotypes: an investigation of type-specific differences in geographic origin and disease.

Because of the nucleotide sequence diversity of different isolates of hepatitis C virus, it has become important to clarify whether distinct genotypes of hepatitis C virus vary with respect to pathogenicity, infectivity, response to antiviral therapy and geographic clustering. We assessed nucleotide sequence variability in the 5' noncoding region of hepatitis C virus, using restriction enzymes to analyze the distribution of hepatitis C virus genotypes, in 80 patients with chronic hepatitis C virus infection. Genotypes were correlated with demographic, clinical and histological features. Thirty-seven patients were infected with type 1, 10 had type 2 and 8 had type 3, and another 23 were infected with a new distinct hepatitis C virus type now classified as type 4. Two were infected with variants whose classification are uncertain. Types 1, 2 and 3 were found in patients from the United Kingdom, southern Europe, Asia, Africa and South America. Nineteen of 23 type 4 genotype isolates were from Middle Eastern patients, compared with 0 of 37 type 1 isolates (p < 0.001). Of 21 Middle Eastern patients, 19 (90.4%) had type 4 hepatitis C virus (p = 0.001, odds ratio = 9). We found no significant difference between the mean ages or mean serum aminotransferase concentrations between the various types. Types 1, 2, 3 and 4 were found in patients with mild-to-moderate disease or severe disease. However, 21 of 29 (72.4%) patients with type 1 who underwent liver biopsy had severe chronic hepatitis, cirrhosis or hepatocellular carcinoma histologically; 8 had mild or moderate chronic hepatitis without cirrhosis (p = 0.03, odds ratio = 2.6).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

[Pathogenic antibodies in systemic lupus erythematosus: anti-LAMP antibodies].

We recently demonstrated that a monoclonal anti-DNA antibody, spontaneously produced in lupus B/W mice, recognizes the same protein(s) at the surface of several human cell types involved in lupus pathogenesis including normal human erythrocytes, normal platelets and rat neuronal tissue. This cell-surface protein(s) cross-react(s) with double-stranded DNA. We suggest to call this protein(s) LAMP [lupus associated membrane protein(s)]. Here we show that: immunoglobulins eluted from kidneys of autoimmune MRL/lpr/lpr mice strongly react with LAMP. Anti-LAMP antibodies are present in large amount in MRL/lpr and B/W mice sera. Anti-LAMP are present in 25 out of 25 human SLE sera ranged as SLE on the basis of revised American Rheumatism Associated classification. Interestingly, two of these sera did not display anti DNA anti-body activity. Taken together, these results strongly suggest a role of LAMP in the pathogeny of SLE.

Animals↗

Genetically-engineered microorganisms: I. Identification, classification, and strain history.

We have argued that accurate identification of the microorganism will form a cornerstone of the assessment of potential hazard. Appropriate methodology for identification exists, and is continually under development and refinement. Organizations such as the American Type Culture Collection will perform certified identifications for relatively low cost. Thus there appears to be little reason that an organism should not be identified insofar as current microbiology allows prior to submission for PMN review. We suggest that a complete microbiological characterization be considered an essential element of an acceptable PMN. To accomplish this, however, current institutional arrangements for the protection of trade secret information needed in the process of identification may need to be improved. An accurate identification of the strain will often provide access to important information with which to evaluate its ecology, pathogenicity, biochemistry, and genetics. Specialized texts, the scientific literature, and professional consultation are ready sources of such information. However, a major effort should be made to establish a data base that can specifically address the needs of biohazard evaluation. This could be done, in part, by collecting information about the construction, and about the behavior in the environment of genetically-engineered microorganisms that are now under development and will soon be tested or used. Identification information may also eventually be useful for the formulation of hypotheses about possible modes of harm or about relative safety, based on phylogenetic relationships. This is a very difficult undertaking at present, however. Microbial taxonomy is currently in a process of radical reevaluation as new macromolecular sequence information reveals previously unsuspected phylogenetic relationships, and disturbs categorizations based on older types of traits such as morphology, etc. This means that both inferences about relative safety and about possible modes of harm from taxonomic relationships must be highly tentative based on current information. Regulatory authorities may wish to consider requesting confirmatory DNA hybridization data or other macromolecular sequence comparisons in cases where strong arguments related to safety must be made from taxonomic information in relatively poorly studied groups of organisms. Detailed strain histories would provide valuable information for safety evaluations.(ABSTRACT TRUNCATED AT 400 WORDS)

Bacteria↗

Zinc-finger transcription factors in plants.

Several classes of zinc-finger motifs are present in transcription factors and function as parts of DNA-binding and protein-protein interaction domains. Most of the known classes of zinc-finger motifs earlier identified in other eucaryotes have also been found in a number of (putative) transcription factors in plants. In addition, some novel classes of zinc fingers have been identified in plants. Many of these proteins have been implicated in the regulation of important biological processes that are unique to plants, such as flower development, light-regulated morphogenesis and pathogen responses. Thus, plants seem to have adopted pre-existing prototype zinc-finger motifs as well as generated new zinc-finger domains to adapt them to various regulatory processes. Detailed analyses of TFIIIA-type plant zinc-finger proteins revealed unique manners of interactions with target DNA sequences, i.e. recognition of spacing, suggesting that plants have developed unique mechanisms even when proto-type functional motifs were adopted. In this review, attempts were made to summarize the current knowledge of (putative) zinc-finger transcription factors according to a structure-based classification, in view of their involvement in specific regulatory processes and interaction with target DNA sequences.

Amino Acid Sequence↗

[Persistence of Chlamydia pneumoniae in coronary plaque tissue. A contribution to infection and immune hypothesis in unstable angina pectoris].

BACKGROUND AND OBJECTIVE: There is an increasing number of pointers towards a causative connection between Chlamydia pneumoniae and atherosclerosis. But the pathogenetic mechanism and intimal structures that are involved remain unclear. Starting with the hypothesis of a chronic infection, as demonstrated by the presence of the chlamydial stress (heat-shock) protein 60 (HSP 60), the presence and localization of these bacterial products in coronary atheromas was investigated. PATIENTS AND METHODS: Coronary atheroma tissue from primary stenoses in 42 patients (36 men, 6 women, mean age 60.2 +/- 7.3 years) was studied immunohistochemically in the course of a retrospective analysis for chlamydial HSP 60. The findings in clinically acute coronary syndrome (Braunwald's classification) present in 27 patients were compared with those in 15 patients with acute angina and evaluated in relation to expression and site of predilection. RESULTS: An immune reaction to chlamydial HSP 60 was demonstrated in 27 of 42 atheromas (64%). Intact, non-atherosclerotic vessels, such as the mammary artery and sphenous vein, showed no such signals. Chlamydial HSP 60 was localized in maximally 23% of all plaque cells, mostly in macrophages/foam cells, more rarely in smooth muscle cells. Chlamydia in foam cells most often revealed ultrastructural patterns that pointed to the persistence of the pathogen. Sites of predilection of chlamydial HSP were predominantly foam cell areas and cell-poor regions, more rarely inflammatory infiltrates and areas of rupture. When comparing both types of lesion, signals for chlamydial HSP 60 were present in 21 of the 27 atheromas (78%) with unstable angina or acute myocardial infarction, but in only 6 of the 15 atheromas (40%) with stable angina. Within the group with the acute coronary syndrome, the prevalence of chronic chlamydial infection was independent of a previous myocardial infarction. CONCLUSIONS: Chlamydial HSP 60 can often be demonstrated in primary coronary stenosis of symptomatic patients. It is most frequently found in macrophages/foam cells and is highly prevalent in the acute coronary syndrome. In-situ findings suggest a pathogenetically relevant role of chronic persistent infection of Chlamydia pneumoniae in unstable coronary plaques.

Adult↗

Severe sepsis and septic shock: defining the clinical problem.

It is generally acknowledged that severe sepsis/septic shock is a major problem in clinical medicine, yet the extent of the problem and its basic immunology remain poorly defined. The generation of accurate statistics about sepsis is confounded by the imprecise and highly variable terminology used to describe sepsis by clinicians around the world. The problem of sepsis is further complicated by the remarkably diverse spectrum of illness encompassed under the term 'sepsis'. Sepsis may range in severity from mild systemic inflammation without significant clinical consequences to multisystem failure in septic shock with an exceedingly high mortality rate. Sepsis connotes a clinical syndrome that may occur in any age group, in markedly different patient populations, and in response to a multitude of microbial pathogens from multiple different anatomical sites within the human body. A concerted effort has been made to standardize definitions of sepsis by the use of international committees and consensus opinions from panels of experts in sepsis research. While consensus definitions of sepsis have proven to be of value, the lack of uniformity in interpretation of these definitions continues to be problematic by clinicians and basic researchers alike. Recently, a new conceptual framework for understanding sepsis has been developed, called the PIRO concept (predisposition, infection, response and organ dysfunction). This has been conceptually modeled from the TNM classification (tumor size, nodal spread, metastases) which has been successfully used in defining treatment and prognostic indicators in clinical oncology. Further refinements in the definitions and predisposing factors of severe sepsis should improve the understanding and management of severe sepsis and septic shock in the near future.

Adult↗

Infections due to Lancefield group F and related Streptococci (S. milleri, S. anginosus).

We can no longer accept classification of streptococci solely on the basis of hemolytic reactions or Lancefield agglutinations. While the "viridans" streptococci cannot be serologically differentiated, physiological separation of the species offers a satisfactory method of classifying human isolates. We review the microbiology of Lancefield group F and related streptococci (S. milleri, S. anginosus), emphasizing microbial ecology and current taxonomic considerations. A series of 28 patients infected with these organisms is presented. There was a striking male predominance in the series (6:1) as well as an obvious association with underlying diseases and/or antecedent trauma. The most remarkable feature of these pathogens is their predilection for abscess formation, confirming their overdue recognition as the most common "viridans" streptococcus recovered from abscesses within internal organs. We observed purulent disease of the nervous and skeletal systems, oral cavity, lung and pleural space, abdomen and subcutaneous tissues. Microbial synergy (i.e. polymicrobic infection) was not a requisite for abscess formation. Four of the five deaths in this series occurred in patients 60 years of age of older. Some degree of variability in antimicrobial susceptibility was noted, so speciation alone may not always provide sufficient information on which to base a therapeutic decision.

Abdomen↗

Phylogeny of certain biocontrol agents with special reference to nematophagous fungi based on RAPd.

A number of phylogenetic studies have been carried out on biocontrol agents having similar biological control activity. However, no work has been carried out to determine the phylogenetic relationship amongst various groups of biological control agents with varied biocontrol properties. Our aim was to derive a phylogenetic relationship between diverse biocontrol agents belonging to the deuteromycetes and determine its correlation with their spore morphology and their biocontrol activity. RAPD was used to assess genomic variability in fungi used as biological control agents which included ten isolates of nematophagous fungi such as Arthrobotrys sp., Duddingtonia sp., Paecilomyces sp. and Verticillium sp., along with two isolates of fungal biocontrol agents such as Trichoderma sp. and two isolates of entomopathogenic fungi including Beauveria sp. A plant pathogenic fungus, Verticillium alboatrum was also included to increase the diversity of Deuteromycetes used. A similarity matrix was created using Jaccard's similarity coefficient & clustering was done using unweighted pair group arithmetic mean method (UPGMA). The final dendogram was created using a combination of two programs, Freetree and TreeExplorer. The phylogenetic tree constructed from the RAPD data showed marked genetic variability among different strains of the same species. The spore morphologies of all these fungi were also studied. The phylogenetic pattern could be correlated with the conidial and conidiophore morphology, a criterion commonly used for the classification of fungi in general and Deuteromycetes in particular. Interestingly, the inferred phylogeny showed no significant grouping based on either their biological control properties or the trapping structures amongst the nematophagous fungi as reported earlier by other workers. The phylogenetic pattern was also similar to the tree obtained by comparing the 18S rRNA sequences from the database. The result clearly indicates that the classical method of classification of these deuteromycete members on the basis of their spore morphology is reliable and could be used for identification of these fungi at species level. The PCR fragment pattern polymorphism exhibited by the various species of a genus and different strains of a species indicates that construction of probes from one or more of these fragments will prove to be useful as a rapid tool for identification of species and strains of nematophagous fungi in future.

Base Sequence↗

Human papillomavirus infection and cervical neoplasia: new perspectives.

This review addresses several problems associated with human papillomavirus (HPV) infection of the cervix and lower female genital tract. These include the definition of HPV infection and its distinction from HPV-associated neoplasia, the distinction of HPV infection from reactive epithelial changes induced by other infections, and the transmission of HPV infection via the male partner. The available evidence indicates that there are two distinct intraepithelial processes in the cervix associated with HPV. One is the classical condyloma and its counterpart in immature epithelium, atypical immature metaplasia. The other is intraepithelial neoplasia, which, like classical infection, may be mature [cervical intraepithelial neoplasia (CIN) with koilocytosis] or immature (high grade CIN or carcinoma in situ). Molecular hybridization studies indicate that HPVs 6 and 11 are most commonly detected in the former, whereas HPVs 16 and 18 DNA are most common in the latter and in invasive cancer. From the clinical standpoint the most important distinction is between HPV-related disease (condyloma or CIN) and reactive changes associated with other pathogens, such as Chlamydia. The former should be removed from the cervix, whereas the latter should be treated medically or followed. It is stressed that therapy should not hinge upon the histological distinction of HPV infection from neoplasia and that all lesions should be removed, by conservative means if possible. This is underscored by the fact that a high proportion of CIN lesions contain areas identical to condyloma and that lesions with deep endocervical canal involvement, including those with features suggesting condyloma, should be treated by cone biopsy to exclude the presence of invasive cancer. Histological classifications for nonneoplastic, HPV-infected, and neoplastic epithelium are proposed. The management of the male partner is still unsettled. However, a large proportion of male partners of these patients have penile lesions and should be included in diagnostic and therapeutic protocols of women with genital HPV infections or neoplasms.

Animals↗

Gastrointestinal lymphoma.

The ability to make treatment recommendations for patients with gastrointestinal lymphoma is hampered by a lack of prospective trials and by a lack of uniformity in classification and staging. Patients with gastric diffuse large B-cell lymphoma have traditionally been treated with surgery and many physicians continue to recommend this approach. However, recent data suggest that these patients can be treated with combination chemotherapy regimens in the same manner as patients with nodal presentations of diffuse large B-cell lymphoma. There is evidence to suggest that adjuvant radiotherapy may improve the outcome for these patients. The recognition that extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue is a distinct clinicopathologic entity and the elucidation of the pathogenic role of Helicobacter pylori has revolutionized the treatment of these gastric lymphomas. Patients with localized disease should be managed with antibiotic therapy initially. Radiation therapy is extremely effective for these patients, but it should probably be reserved for patients who fail anti-H. pylori treatment.

Adult↗