[Computed tomographic features of coronavirus disease-2019 (COVID-19) pneumonia in three groups of Iranian patients: A single center study]. / Características en tomografía computarizada de la neumonía por coronavirus-2019 (COVID-19) en tres grupos de pacientes iraníes: estudio de un solo centro.

Introduction and Objectives: The pivotal role of chest computed tomographic (CT) to diagnosis and prognosis coronavirus disease-2019 (COVID-19) is still an open field to be explored. This study was conducted to assess the CT features in confirmed cases with COVID-19. Materials and Methods: Retrospectively, initial chest CT data of 363 confirmed cases with COVID-19 were reviewed. All subjects were stratified into three groups based on patients' clinical outcomes; non-critical group (n=194), critical group (n=65), and death group (n=104). The detailed of CT findings were collected from patients' medical records and then evaluated for each group. In addition, multinomial logistic regression was used to analyze risk factors according to CT findings in three groups of patients with COVID-19. Results: Compared with the non-critical group, mixed ground-glass opacities (GGO) and consolidation lesion, pleural effusion lesion, presence of diffuse opacity in cases, more than 2 lobes involved and opacity scores were significantly higher in the critical and death groups (P<0.05). Having more mixed GGO with consolidation, pleural effusion, lack of pure GGO, more diffuse opacity, involvement of more than 2 lobes and high opacity score identified as independent risk factors of critical and death groups. Conclusion: CT images of non-critical, critical and death groups with COVID-19 had definite characteristics. CT examination plays a vital role in managing the current COVID-19 outbreak, for early detection of COVID-19 pneumonia. In addition, initial CT findings may be useful to stratify patients, which have a potentially important utility in the current global medical situation.

Distribution and phylogenetic analysis of Blastocystis sp. subtypes isolated from IBD patients and healthy individuals in Iran.

Blastocystis is a single-celled intestinal parasite commonly found in humans and a broad range of animals all over the world. In humans, its role in health and disease remains unsettled. The aim of our study was to investigate the distribution of Blastocystis and Blastocystis subtypes (ST) in patients with inflammatory bowel disease (IBD) and control subjects. A total of 71 stool samples were collected from IBD patients, 69 and 2 of whom had ulcerative colitis (UC) and Crohn's Disease (CD), respectively. Moreover, 166 stool samples from healthy subjects were included as control samples. All stool samples were cultivated, and 550-bp fragments of the small subunit ribosomal RNA gene was amplified from Blastocystis-positive cultures. All PCR-positive samples were sequenced. Blastocystis was observed in 9 (12.67%) and 35 (21.1%) IBD patients and healthy controls, respectively. There was no statistically significant correlation between IBD and presence of Blastocystis (P = 0.147). There was a statistically significant correlation between age and Blastocystis colonization in the IBD group (P < 0.05), but not among healthy controls. No significant correlation between gender and colonization was observed. ST1 and ST3 were obtained from 1 (12.5%) and 7 (87.5%) IBD patients, respectively, while in the healthy control group, subtypes 1, 2, and 3 were found in 14 (40%), 12 (34.28%), and 9 (25.72%), respectively. Phylogenetic analysis showed no variation in the distribution of subtypes nor intra-subtype genetic diversity between samples acquired from IBD patients and healthy controls. This study showed a trend towards a lower prevalence of Blastocystis in IBD patients than in control subjects. ST3 sequences isolated from IBD patients and control individuals did not appear to differ genetically.

The first study on opportunistic intestinal microsporidiosis in IBD patients receiving immunosuppressive medications in Iran.

Microsporida are known as opportunistic unicellular organisms and have recently been reclassified as fungi that have been frequently reported from patients with congenital and acquired immunity failure disorders, worldwide. However, use of immunosuppressive medications in inflammatory bowel disease (IBD) patients significantly decreases overall immunity, and increases their susceptibility to opportunistic infections. Totally, 71 stool samples were collected from IBD patients consisted of 69 ulcerative colitis (UC) patients and two Crohn's disease (CD) patients. All patients had taken immunosuppressive and/or immunomodulator drugs for at least 3 weeks. DNA was extracted from all stool samples and Nested PCR was performed using genus-specific primers based on small subunit ribosomal RNA (SSU rRNA) gene. Fisher's Exact Test was applied to evaluate statistical association between microsporidia infection and sex, age and types of IBD. Mean of age ± s.d., women and men percentage of the attended patients were 36·17 ± 11·93, 60·6%, and 39·4%, respectively. A 440-bp fragment of SSU rRNA gene attributed to Enterocytozoon bieneusi was amplified from 12·7% of IBD patients. No Encephalitozoon DNA was detected in the samples. No microsporidia-positive sample was found in CD patients. Fisher's Exact Test showed that there was no statistically significant correlation between intestinal microsporidiosis and age, sex, and IBD types with P values: 0·389, 1·00, and 1·00, respectively. This study has shown IBD patients undergoing immunosuppressive/immunomodulators medications, which may be susceptible to intestinal microsporida infection. E. bieneusi is the commonest intestinal microsporidan reported from IBD patients.

Association of diverse bacterial communities in human bile samples with biliary tract disorders: a survey using culture and polymerase chain reaction-denaturing gradient gel electrophoresis methods.

Bacterial infection is considered a predisposing factor for disorders of the biliary tract. This study aimed to determine the diversity of bacterial communities in bile samples and their involvement in the occurrence of biliary tract diseases. A total of 102 bile samples were collected during endoscopic retrograde cholangiopancreatography (ERCP). Characterization of bacteria was done using culture and polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) methods. Antimicrobial susceptibility of the isolates was determined based on the Clinical and Laboratory Standards Institute (CLSI) guidelines and identity of the nucleotide sequences of differentiated bands from the DGGE gels was determined based on GenBank data. In total, 41.2 % (42/102) of the patients showed bacterial infection in their bile samples. This infection was detected in 21 % (4/19), 45.4 % (5/11), 53.5 % (15/28), and 54.5 % (24/44) of patients with common bile duct stone, microlithiasis, malignancy, and gallbladder stone, respectively. Escherichia coli showed a significant association with gallstones. Polymicrobial infection was detected in 48 % of the patients. While results of the culture method established coexistence of biofilm-forming bacteria (Pseudomonas aeruginosa, E. coli, Klebsiella pneumoniae, Enterococcus spp., and Acinetobacter spp.) in different combinations, the presence of Capnocytophaga spp., Lactococcus spp., Bacillus spp., Staphylococcus haemolyticus, Enterobacter or Citrobacter spp., Morganella spp., Salmonella spp., and Helicobacter pylori was also characterized in these samples by the PCR-DGGE method. Multidrug resistance phenotypes (87.5 %) and resistance to third- and fourth-generation cephalosporins and quinolones were common in these strains, which could evolve through their selection by bile components. Ability for biofilm formation seems to be a need for polymicrobial infection in this organ.

Association of CagPAI integrity with severeness of Helicobacter pylori infection in patients with gastritis.

BACKGROUND AND AIM: The Helicobacter pylori cag pathogenicity island (cagPAI) is involved in delivery of CagA effector protein and peptidoglycan into host cells and also in IL-8 induction in the human gastric tissue. Diversity of cagPAI may affect disease status and clinical outcome of the infected patients. Our study was aimed to investigate diversity of this island and its intactness in Iranian patients to investigate possible associations between cagPAI integrity and pathological changes of the infected tissue. MATERIAL/PATIENTS AND METHODS: Out of the 75 patients, H. pylori strains were obtained from 30 patients with severe active gastritis (SAG) (n=11), moderate chronic gastritis (CG) (n=14) and intestinal metaplasia/dysplasia (IM) (n=5). Intactness of the cagPAI was determined using 12 sets of primer pairs specific for functionally important loci of cagPAI by polymerase chain reaction (PCR). RESULTS: The cagPAI positive strains were significantly observed in patients with SAG (52.4%) in comparison to those presenting CG (33.3%) and IM (14.3%). In addition, the presence of intact cagPAI was 87.5% in H. pylori strains isolated from patients with SAG, which was higher than those obtained from patients with CG (12.5%) or IM (0%). A significant increase in the frequency of cagα-cagY and cagW-cagT segments, as exterior proteins of the CagPAI, was illustrated in strains from SAG patients compared with those from patients with CG. CONCLUSIONS: Overall, these results strongly proposed an association between the severity of histopathological changes and intactness of cagPAI in the gastric tissue of patients infected with H. pylori.

Celiac disease and dysfunctional uterine bleeding; the efficiency of gluten free diet.

The aim of this study was to investigate the relation between Celiac disease (CD) and unexplained dysfunctional uterine bleeding (DUB) in celiac women. The celiac patients were selected from women who were referred to celiac department. Controls were selected from those women without any signs of celiac disease and matched with age. Meanwhile, a trained physician was ready to explain the study, and then in case of their allowance, a questionnaire was completed by the physician. 24 % of celiac women reported a past history of at least one menstrual cycle disorder vs 10 % of controls reported these problems (p=0.038) and higher percentage of unexplained DUB has been observed in celiac women. All celiac patients were undertaking gluten free diet for at least 3 months and the celiac patients who reported the history of DUB were again interviewed for any signs of unexplained DUB. From 12 celiac women with DUB, 10 patients reported no more unexplained DUB after getting gluten-free diet (83.3 %). The occurrence of a significant correlation between CD and DUB suggests the possibility of considering CD as one of the potential causes of abnormal uterine bleeding. Therefore, celiac disease must be seriously considered in the screening of patients with reproductive disorders (Tab. 2,Ref. 23).

Application of artificial intelligence techniques for non-alcoholic fatty liver disease diagnosis: A systematic review (2005-2023).

BACKGROUND AND OBJECTIVES: Non-alcoholic fatty liver disease (NAFLD) is a common liver disease with a rapidly growing incidence worldwide. For prognostication and therapeutic decisions, it is important to distinguish the pathological stages of NAFLD: steatosis, steatohepatitis, and liver fibrosis, which are definitively diagnosed on invasive biopsy. Non-invasive ultrasound (US) imaging, including US elastography technique, and clinical parameters can be used to diagnose and grade NAFLD and its complications. Artificial intelligence (AI) is increasingly being harnessed for developing NAFLD diagnostic models based on clinical, biomarker, or imaging data. In this work, we systemically reviewed the literature for AI-enabled NAFLD diagnostic models based on US (including elastography) and clinical (including serological) data. METHODS: We performed a comprehensive search on Google Scholar, Scopus, and PubMed search engines for articles published between January 2005 and June 2023 related to AI models for NAFLD diagnosis based on US and/or clinical parameters using the following search terms: "non-alcoholic fatty liver disease", "non-alcoholic steatohepatitis", "deep learning", "machine learning", "artificial intelligence", "ultrasound imaging", "sonography", "clinical information". RESULTS: We reviewed 64 published models that used either US (including elastography) or clinical data input to detect the presence of NAFLD, non-alcoholic steatohepatitis, and/or fibrosis, and in some cases, the severity of steatosis, inflammation, and/or fibrosis as well. The performances of the published models were summarized, and stratified by data input and algorithms used, which could be broadly divided into machine and deep learning approaches. CONCLUSION: AI models based on US imaging and clinical data can reliably detect NAFLD and its complications, thereby reducing diagnostic costs and the need for invasive liver biopsy. The models offer advantages of efficiency, accuracy, and accessibility, and serve as virtual assistants for specialists to accelerate disease diagnosis and reduce treatment costs for patients and healthcare systems.

Genetic variability of Blastocystis sp. isolated from symptomatic and asymptomatic individuals in Iran.

Blastocystis is an unusual enteric protozoan parasite of humans and many animals whose pathogenic potential is still controversial. To increase the understanding of the molecular epidemiology of this emerging parasite and due to its potential impact on public health, its subtypes (STs) in Iranian symptomatic and asymptomatic individuals were determined. A total of 100 Blastocystis isolates by microscopy and culture methods were obtained. DNA was extracted from the positive culture isolates, and the Blastocystis subtypes were identified using seven subtype-specific sequenced-tagged site (STS) primers. Four subtypes, ST3 as dominant (53 %), followed by ST1 (48 %), ST5 (33 %), and ST2 (7 %) were identified. In this study, ST1 in gastrointestinal patients compared to asymptomatic individuals was significantly dominant (p = 0.001). From 33 (33 %) mixed subtype infections, ST1, 3 (14 %) was significantly related to GI symptoms (p = 0.045), and eight mixed infections with three different STs, which are under reported, were also identified.

Molecular identification of animal isolates of Echinococcus granulosus from Iran using four mitochondrial genes.

Mitochondrial genes have more power than nuclear genes in reconstructing phylogenetic relationships among closely related species because of their faster sequence evolution. The aim of this study was to use the complete or near-complete sequences from three mitochondrial genes (cox1, nad1 and atp6) and partial sequences of the 12S rRNA gene to infer relationships among isolates of Echinococcus granulosus from Iran. Two hundred and twenty-nine isolates of E. granulosus were collected from cattle, camels, sheep, buffalo and goats from different geographical areas. Most individuals were found to possess the G1 genotype but some of the camel samples belonged to the G6 genotype. Newly designed primers for cox1, nad1 and atp6 genes amplified bands of 1830, 708 and 1157 bp for the G1 genotype and 1856, 705, 1054 bp for the G6 genotype, respectively. The result of this survey showed that atp6 and nad1 genes are good molecular markers for identifying E. granulosus isolates from a range of hosts in Iran.

Computed tomographic features of coronavirus disease-2019 (COVID-19) pneumonia in three groups of Iranian patients: A single center study.

INTRODUCTION AND OBJECTIVES: The pivotal role of chest computed tomographic (CT) to diagnosis and prognosis coronavirus disease-2019 (COVID-19) is still an open field to be explored. This study was conducted to assess the CT features in confirmed cases with COVID-19. MATERIALS AND METHODS: Retrospectively, initial chest CT data of 363 confirmed cases with COVID-19 were reviewed. All subjects were stratified into three groups based on patients' clinical outcomes; non-critical group (n=194), critical group (n=65), and death group (n=104). The detailed of CT findings were collected from patients' medical records and then evaluated for each group. In addition, multinomial logistic regression was used to analyze risk factors according to CT findings in three groups of patients with COVID-19. RESULTS: Compared with the non-critical group, mixed ground-glass opacities (GGO) and consolidation lesion, pleural effusion lesion, presence of diffuse opacity in cases, more than 2 lobes involved and opacity scores were significantly higher in the critical and death groups (P<0.05). Having more mixed GGO with consolidation, pleural effusion, lack of pure GGO, more diffuse opacity, involvement of more than 2 lobes and high opacity score identified as independent risk factors of critical and death groups. CONCLUSION: CT images of non-critical, critical and death groups with COVID-19 had definite characteristics. CT examination plays a vital role in managing the current COVID-19 outbreak, for early detection of COVID-19 pneumonia. In addition, initial CT findings may be useful to stratify patients, which have a potentially important utility in the current global medical situation.

Sentinel hospital-based surveillance of rotavirus diarrhea in iran.

BACKGROUND: Rotavirus is the most common causes of severe, acute diarrhea during childhood and is an important cause of morbidity and mortality in developing countries. We established active hospital-based surveillance of childhood diarrhea to assess the scope of severe rotavirus disease in Iran. METHODS: From May 2006 through April 2007, prospective surveillance of rotavirus diarrhea among children aged <5 years was conducted in 5 sentinel hospitals in Iran. Stool samples were tested for rotavirus using a commercially available enzyme immunoassay, and rotavirus-positive samples were genotyped using reverse-transcriptase polymerase chain reaction. RESULTS: Of 2198 children admitted to the hospital for acute gastroenteritis, 1298 (59.1%) had stool samples test positive for rotavirus by enzyme immunoassay. Of the rotavirus episodes, 85% occurred during the first 2 years of life, with the peak prevalence of severe rotavirus disease occurring from September through January. Among the 110 rotavirus-positive samples that were genotyped, G4P[8] was the most commonly detected rotavirus genotype (30.9% of strains). Other commonly detected genotypes included P[8] with G nontypeable (21.8%), G4 with P nontypeable (13.6%), G1[P8] (10.9%), and G2[P4] (5.5%). CONCLUSIONS: Rotavirus is the most common cause of severe diarrhea in Iran, which indicates that safe and effective rotavirus vaccination in Iran is a public health priority.

A 3'-untranslated region variant (rs2289046) of insulin receptor substrate 2 gene is associated with susceptibility to nonalcoholic fatty liver disease.

PURPOSE: Nonalcoholic fatty liver disease (NAFLD) is an increasing global health concern defined by excessive hepatic fat content in the absence of excessive alcohol consumption. Regarding the key role of insulin and insulin resistance in NAFLD, we investigated whether insulin receptor substrate 1 (IRS1) and insulin receptor substrate 2 (IRS2) gene variants were associated with NAFLD risk. METHODS: In this case-control study, 305 subjects including 151 cases with biopsy-proven NAFLD and 154 controls were enrolled. All the subjects were genotyped for IRS1 (rs1801278) and IRS2 (rs2289046) gene variants using PCR-RFLP method. RESULTS: Our findings showed that the IRS2 rs2289046 "GG+AG" genotype compared with "AA" genotype to be a marker of decreased NAFLD susceptibility and the difference remained significant even after adjustment for confounding factors including age, BMI, sex, smoking status, systolic blood pressure, and diastolic blood pressure (P=0.014; OR=0.50, 95%CI= 0.29-0.87). Furthermore, the IRS2 "G" allele was significantly underrepresented in the cases with NAFLD than controls (P=0.026 ; OR=0.62, 95%CI=0.41-0.94). However, no significant difference was found for IRS1 rs1801278 gene variant. CONCLUSIONS: This study suggests, for the first time, that the IRS2 gene rs2289046 variant may play a role in NAFLD susceptibility. Nevertheless, this observation warrants further investigations in other populations.

The association of vacA genotypes and Helicobacter pylori-related gastroduodenal diseases in the Middle East.

The variations in the three regions of the Helicobacter pylori vacA gene, the signal (s1 and s2), intermediate (i1 and i2) and middle regions (m1 and m2), are known to cause the differences in vacuolating activities. However, it was unclear whether these vacA genotypes are associated with the development of gastric cancer and peptic ulcer in the Middle East. The aim was to identify the prevalence of vacA genotypes in the Middle East and the association with gastroduodenal diseases. We investigated the relationship of vacA genotypes to H. pylori-related disease development by meta-analysis using previous reports of 1,646 patients from the Middle East. The frequency of the vacA s1, m1 and i1 genotypes in the Middle Eastern strains was 71.5% (1,007/1,409), 32.8% (427/1,300) and 40.7% (59/145), respectively. Importantly, the frequency of vacA s- and m-region genotypes significantly differed between the north and south parts of the Middle East countries (P < 0.001). The vacA genotypes significantly increased the risk of gastric cancer (odds ratio [OR]: 4.02, 95% confidence interval [CI]: 1.98-8.14 for the s1 genotype; 2.50, 1.62-3.85 for m1; 5.27, 1.97-14.1 for s1m1; 15.03, 4.69-48.17 for i1) and peptic ulcers (OR: 3.07, 95% CI: 2.08-4.52 for s1; 1.81, 1.36-2.42 for m1). The cagA-positive genotype frequently coincided with the s1, m1 and i1 genotypes. The vacA s- and m-region genotypes may be useful risk factors for gastrointestinal diseases in the Middle East, similar to European and American countries. Further studies will be required to evaluate the effects of the i-region genotype.

Prognostic factors in gastric cancer using log-normal censored regression model.

BACKGROUND & OBJECTIVE: Gastric cancer is one of the most common cancers in the world. It is rarely detected early, and the prognosis remains poor. Cox proportional hazard model is used to examine the relationship between survival and covariates. Parametric survival models such as log normal regression model can also be used for this analysis. We used log normal regression model in this study to evaluate prognostic factors in gastric cancer and compared with Cox model. METHODS: We retrospectively studied the 746 patients diagnosed with gastric cancer admitted in a referral hospital in Tehran, Iran, from February 2003 through January 2007. Age at diagnosis, sex, extent of wall penetration, histology type, tumour grade, tumour size, pathologic stage, lymph node metastasis and presence of metastasis were entered into a log normal model. Hazard rate (HR) was employed to interpret the risk of death and the results were compared with Cox regression. The AIC (Akaike Information Criterion) was employed to compare the efficiency of models. RESULTS: Univariate analysis indicated that with increasing age the risk of death increased significantly in both log normal and Cox models. Patients with greater tumour size were also in higher risk of death followed by those with poorly differentiated and moderately differentiated in tumour grade and advanced pathologic stage. The presence of metastasis was significant prognostic factor only in log normal analysis. In final multivariate model, age was still a significant prognostic factor in Cox regression but it was not significant in log normal model. Presence of metastasis followed by histology type were other prognostic features found significant in log normal results. Based on AIC, log normal model performed better than Cox. INTERPRETATION & CONCLUSION: Our results suggest that early detection of patients in younger age and in primary stages and grade of tumour could be important to decrease the risk of death in patients with gastric cancer. Comparison between Cox and log normal models indicated that log normal regression model can be a useful statistical model to find prognostic factors instead of Cox.

Celiac disease and Hp infection association in Iran.

BACKGROUND: we assessed the prevalence, the related symptoms, and the endoscopic and histologic gastric features of celiac disease (CD) in patients with Helicobacter pylori (Hp). METHODS: 450 dyspeptic patients were studied. Biopsies of gastric antrum and duodenum, CD serology, and total IgA were obtained. Histological findings were scored with the Marsh-Rostami criteria. RESULTS: 411 (91.3%) patients were Hp positive. Duodenal histology was normal in 385 (85.6%) patients, 124 (27.5%) had duodenitis and 28 (6.2%) showed duodenal abnormalities (Marsh I-IIIc). Twenty three/28 (82.1%) patients with malabsorption pattern were also Hp positive. Serological analysis: 12 of 31 (38.7%) positive patients had abnormal histology (Marsh I,-IIIc). Nine out 450 patients were IgA deficient; none of them was serologically positive for CD. CONCLUSION: although a high prevalence of Hp infection was found in this study, the relationship between Hp infection and CD was similar to that reported in other geographic areas.

Seroprevalence of Helicobacter pylori in Nahavand: a population-based study.

In this cross-sectional study, we evaluated H. pylori seroprevalence and the relevant factors in 1518 people aged > or = 6 years from the general population of Nahavand, western Islamic Republic of Iran. Questionnaires covering sociodemographic variables were completed by interview. Blood samples were taken from each individual. Sera were tested for anti-H. pylori IgG using commercial enzyme immunoassay. Overall, seroprevalence of H. pylori was high, 71.0% (95% CI: 69.0%-73.0%). There was a gradual increase with age. Based on multivariate adjustment, only female sex and age could be considered risk factors.

Echinococcus granulosus strain differentiation in Iran based on sequence heterogeneity in the mitochondrial 12S rRNA gene.

Parasite strain characterization is essential for the establishment of a prevention and control strategy in any endemic area. The aim of this study was to characterize different Echinococcus granulosus isolates from Iran by using DNA sequences of the mitochondrial 12S rRNA gene. Thirty livers and lungs of cattle, sheep and goats naturally infected with E. granulosus were collected from abattoirs in northern and western Iran between June and October 2007. These samples yielded 18 fertile cysts which we used for the genetic work. We designed and tested two new primer pairs which specifically amplify portions of the mitochondrial 12S rRNA gene of the two strains (G1 and G6) of E. granulosus known to occur in Iran. One primer pair amplified a fragment of 259 base pairs (bp) from only the G1 strain. The second pair amplified a fragment of 676 bp from the G6 strain. The G1 genotype was identified in all fertile cyst samples, in agreement with previous studies in Iran. Ten of our samples and a single reference sample of the G6 strain were sequenced and compared with the G1 and G6 sequences deposited in GenBank.

Seroprevalence of hepatitis E in Nahavand, Islamic Republic of Iran: a population-based study.

A 2-month community-based survey in February/March of 2003 was carried out to study the seroprevalence of hepatitis E virus (HEV) infection in Nahavand, Islamic Republic of Iran. From each of 6 urban regions of Nahavand, 304 inhabitants > or = 6 years were recruited through systematic random sampling (total 1824). Participants were tested for anti-HEV IgG using ELISA. The overall seroprevalence of HEV was 9.3% (95% CI: 8.2%-10.9%). Based on multivariate adjustment, only sex (OR = 1.61, 95% CI: 1.13-2.28) and age (OR = 1.03, 95% CI: 1.01-1.04) emerged as significant risk factors. This intermediate prevalence urges further investigations on HEV infection in the Islamic Republic of Iran.

Immunohistochemical analysis of p53, cyclinD1, RB1, c-fos and N-ras gene expression in hepatocellular carcinoma in Iran.

AIM: To study the effect of some genes especially those involved in cell cycle regulation on hepatocellular carcinoma. METHODS: Paraffin-embedded tissue samples of 25 patients (18 males and 7 females) with hepatocellular carcinoma were collected from 22 pathology centers in Tehran during 2000-2001, and stained using immunohistochemistry method (avidin-biotin-peroxidase) for detection of p53, cyclinD1, RB1, c-fos and N-ras proteins. RESULTS: Six (24%), 5 (20%), 12 (48%) and 2 samples (8%) were positive for p53, cyclinD1, C-fos and N-ras expression, respectively. Twenty-two (88%) samples had alterations in the G1 cell-cycle checkpoint protein expression (RB1 or cyclinD1). P53 positive samples showed a higher (9 times) risk of being positive for RB1 protein than p53 negative samples. Loss of expression of RB1 in association with p53 over-expression was observed in 4 (66.7%) of 6 samples. Loss of expression of RB1 was seen in all cyclinD1 positive, 20 (90.9%) N-ras negative, and 11 (50%) C-fos positive samples, respectively. CyclinD1 positive samples showed a higher (2.85 and 4.75 times) risk of being positive for c-fos and N-ras expression than cyclinD1 negative samples. CONCLUSION: The expression of p53, RB1 and c-fos genes appears to have a key role in the pathogenesis of hepatocellular carcinoma in Iran. Simultaneous overexpression of these genes is significantly associated with their loss of expression during development of hepatocellular carcinoma.