Matrix metalloproteinase­1 and microRNA­486­5p in urinary exosomes can be used to detect early lung cancer: A preliminary report.

The present study describes a novel molecular-genetic method suitable for lung cancer (LC) screening in the work-place and at community health centers. Using urinary-isolated exosomes from 35 patients with LC and 40 healthy volunteers, the expression ratio of MMP-1/CD63, and the relative expression levels of both microRNA (miRNA)-21 and miRNA-486-5p were measured. MMP-1/CD63 expression ratio was significantly higher in patients with LC than in the healthy controls {1.342 [95% confidence interval (CI): 0.890-1.974] vs. 0.600 (0.490-0.900); P<0.0001}. The relative expression of miRNA-486-5p in male healthy controls was significantly different from that in female healthy controls, whereas there was no significant difference in miRNA-21. Receiver operating characteristic curve (ROC) analysis of MMP-1/CD63 showed 92.5% sensitivity and 54.3% specificity, whereas miRNA-486-5p showed 85% sensitivity and 70.8% specificity for men, and 70.0% sensitivity and 72.7% specificity for women. The logistic regression model used to evaluate the association of LC with the combination of MMP-1/CD63 and miRNA-486-5p revealed that the area under the ROC curve was 0.954 (95% CI: 0.908-1.000), and the model had 89% sensitivity and 88% specificity after adjusting for age, sex and smoking status. These data suggested that the combined analysis of MMP-1/CD63 and miRNA-486-5p in urinary exosomes may be used to detect patients with early-stage LC in the work-place and at community health centers, although confirmational studies are warranted.

Palivizumab prophylaxis in preterm infants and subsequent wheezing/asthma: 10-year follow-up study.

BACKGROUND: Respiratory syncytial virus (RSV) causes not only infantile recurrent wheezing but also the development of asthma. To investigate whether palivizumab, an anti-RSV monoclonal antibody, prophylaxis given to preterm infants during the first RSV season reduces the incidence of subsequent recurrent wheezing and/or development of asthma, at 10 years of age. METHODS: We conducted an observational prospective multicenter (52 registered hospitals in Japan) case-control study in preterm infants with a gestational age between 33 and 35 weeks followed for 6 years. During the 2007-2008 RSV season, the decision to administer palivizumab was made based on standard medical practice (SCELIA study). Here, we followed these subjects until 10 years of age. Parents of study subjects reported the patients' physician's assessment of recurrent wheezing/asthma, using a report card and a novel mobile phone-based reporting system using the internet. The relationship between RSV infection and asthma development, as well as the relationship between other factors and asthma development, were investigated. RESULTS: Of 154 preterm infants enrolled, 113 received palivizumab during the first year of life. At 10 years, although both recurrent wheezing and development of asthma were not significantly different between the treated and untreated groups, maternal smoking with aeroallergen sensitization of the patients was significantly correlated with physician-diagnosed asthma. CONCLUSIONS: In contrast to the prior study results at 6 years, by 10 years palivizumab prophylaxis had no impact on recurrent wheezing or asthma, but there was a significant correlation between maternal passive smoking with aeroallergen sensitization and development of asthma by 10 years of age.

Shoulder function after neck dissection with level IIb preservation: a prospective observational study.

BACKGROUND: Shoulder syndrome can be reduced by preserving the spinal accessory nerve (SAN). However, it is unclear whether performing level IIb preservation will decrease the risk of SAN injury and shoulder syndrome. AIMS/OBJECTIVES: We investigated whether neck dissection with level IIb preservation can reduce shoulder dysfunction and postoperative quality of life (QOL) in head and neck cancer patients. MATERIAL AND METHODS: This prospective observational study enrolled patients who underwent neck dissection from 2011 to 2014. Patients were divided into three groups (level IIb preservation group [group 1], IIb dissection group [group 2], and IIb and V dissection group [group 3]). Postoperative shoulder function and QOL were evaluated among the three groups. RESULTS: There were a total of 35 neck sides in three groups, with nine neck sides in group 1, 16 neck sides in group 2, and 10 neck sides in group 3. Although the results showed less shoulder dysfunction in group 1 at early postoperative period. The QOL in group 1 was preserved in the early postoperative period. CONCLUSIONS AND SIGNIFICANCE: Neck dissection with level IIb preservation may help reduce shoulder syndrome and maintain QOL in the early postoperative period.

Descriptive study of chest x-ray examination in mandatory annual health examinations at the workplace in Japan.

The utility of chest x-ray examination (CXR) in mandatory annual health examinations for occupational health is debatable in Japan. This study aimed to provide basic data to consider future policies for mandatory annual health examinations in the workplace. A nationwide descriptive survey was performed to determine the rate of detection of tuberculosis, lung cancer, and other diseases through CXR in organizations associated with National Federation of Industrial Health Association. The rate of finding on CXR conducted during annual health examinations in FY2016 was evaluated. Data regarding diagnosis based on follow-up examination findings were obtained and compared with the national statistics. In addition, CXR findings were compared with the results of low-dose lung computed tomography performed at the Hitachi Health Care Center. From 121 surveyed institutions, 88 institutions with 8,669,403 workers were included. For all ages, 1.0% of examinees required follow-up examination. Among 4,764,985 workers with diagnosis data, the tuberculosis detection rate was 1.8-5.3 per 100,000 persons. For Lung cancer, 3,688,396 workers were surveyed, and 334 positive cases were detected. The lung cancer detection rate using CXR was 9.1-24.4 per 100,000 persons. From 164 cases with information regarding the clinical stage, 72 (43.9%) had Stage I lung cancer. From 40,045 workers who underwent low-dose computed tomography multiple times, 31 lung cancer cases, all with Stage I disease, were detected (detection rate: 77.4 per 100,000 persons). Our findings suggest that CXR plays a little role in the detection of active tuberculosis. With regard to LC screening, the detection rate of LC by CXR was lower, approximately 50%, than the expected rate (41.0 per 100,000 persons) of LC morbidity based on the age-sex distribution of this study population. However, the role of CXR for LC screening cannot be mentioned based on this result, because assessment of mortality reduction is essential to evaluate the role.

Evaluation of Erastin as a Therapeutic Agent Under Hypoxic Conditions in Pancreatic Cancer Cells.

BACKGROUND/AIM: In pancreatic cancer tissues, hypoxic areas exist due to poor blood flow. Attenuation of the pharmacological efficacy of existing anticancer drugs in these hypoxic areas necessitates the search for novel anticancer compounds. We aimed to determine whether erastin exhibits anticancer effects in a hypoxic environment. MATERIALS AND METHODS: Pancreatic cancer cell lines were subjected to cobalt chloride, a hypoxia-mimicking agent. Cell viability assay, measurement of reactive oxygen species, and western blotting analysis were conducted to investigate the efficacy of erastin under hypoxic environments. RESULTS: Erastin exhibited remarkable cytotoxicity and induced apoptosis under hypoxic conditions. Furthermore, erastin triggered the intracellular accumulation of reactive oxygen species in a hypoxic environment. Subsequent treatment with N-acetylcysteine, an antioxidant, markedly attenuated cytotoxicity, and apoptosis. CONCLUSION: Erastin induces cell death by accumulation of intracellular reactive oxygen species and inducing apoptosis under hypoxic conditions, proving its potential for further development as a novel anticancer compound.

A nationwide cross-sectional survey on hepatitis B and C screening among workers in Japan.

In Japan, there is no publicly funded screening for hepatitis B virus (HBV) and hepatitis C virus (HCV) infections (using HBs antigen and HCV antibody, respectively) among workers, and workplace health programmes play a crucial role in reducing viral hepatitis-related deaths. The national number of hepatitis screening tests conducted in the workplace is unknown. To provide baseline data for policy formulation, we conducted a nationwide survey to estimate these parameters using data from approximately 10.5 million workers (6.8 million men and 3.8 million women) who underwent mandatory health examinations in their workplaces between April 2016 and March 2017. Among these workers, 494,303 (5.23%, 95% confidence interval [CI] 5.22%-5.24%) and 313, 193 (3.82%, 95% CI 3.81%-3.84%) were screened for HBV and HCV, respectively. Among those who were screened, 0.28% (95% CI 0.27-0.30%) and 0.35% (95% CI 0.33-0.37%) tested positive for HBs antigen and HCV antibody, respectively. According to the age-specific prevalence from the survey an estimated 0.30 and 0.14 million workers in Japan require treatment for HBV and HCV, respectively. To reduce viral hepatitis-related deaths by efficiently identifying workers who need treatment and promoting access to treatment, one-time hepatitis screening of all workers should be considered.

Index extensor digitorum communis tendon entrapment in a growth plate injury of distal radius.

BACKGROUND: We report a rare case in which an epiphyseal injury of the distal radius caused entrapment and rupture of the index extensor digitorum communis tendon during conservative treatment. CASE PRESENTATION: A 16-year-old girl suffered epiphyseal injury of the right distal radius with palmar displacement during playing judo. Closed reduction and application of a short arm cast were performed. 4 weeks after injury, movement of her index finger was limited. Extensor tendons adhesion at the fracture site was suspected. It was found that the index extensor digitorum communis tendon had become trapped and ruptured at the site of epiphyseal injury, then reconstruction was done by tendon transfer. At 12 months after surgery, the patient had resumed judo at the same level as before the injury. CONCLUSIONS: The present case suggested that there is a risk of extensor tendon entrapment associated with closed reduction.

Characteristics of breath sound in infants with risk factors for asthma development.

BACKGROUND: Breath sound parameters have been suggested as biomarkers of the airway narrowing in children. Using a commercially available breath sound analyzer, the characteristics of the airway condition were investigated in infants with the risk factors for asthma development. METHODS: A total of 443 infants (mean age, 9.9 months; range, 3-24 months) were included in the present study. The breath sound parameters of the frequency limiting 99% of the power spectrum (F99), the roll-off from 600 to 1200 Hz (Slope) and spectrum curve indices, the total area under the curve of the dBm data (A3/AT) and the ratio of power and frequency at 50% and 75% of the highest frequency of the power spectrum (RPF75 and RPF50), were evaluated. Using an ATS-DLD based original Japanese questionnaire, we examined the characteristics of airway condition of infants. RESULTS: Finally, 283 infants in good health were included in the present study. The RPF75, RPF50, Slope and F99 in infants with positive results of allergy and atopic dermatitis were significantly increased more than those in the infants with negative result. CONCLUSIONS: Our data highlight the characteristics of breath sounds in infants with risk factors for asthma. The breath sound analysis may be useful for assessing the airways of infants for asthma development.

Verification of the International Society of Urological Pathology recommendations in Japanese patients with clear cell renal cell carcinoma.

The aim of the present study was to evaluate the validity of potential prognostic parameters of clear cell renal cell carcinoma (ccRCC) recommended by the 2012 International Society of Urological Pathology (ISUP) Consensus Conference in the Japanese population. We reviewed 406 Japanese patients with localized or locally advanced ccRCC who underwent curative surgery during 2004-2014 at Tokai University Hospital (Isehara, Japan) and were followed up for >2 years after surgery. A single pathologist reviewed all the histological slides. Morphological subtype and pathological T stage were reassigned according to the 2016 World Health Organization and TNM classifications. Sarcomatoid differentiation (SD), rhabdoid differentiation (RD), tumor necrosis (TN) and microvascular invasion (MVI) were assessed according to the 2012 ISUP recommendations. Nuclear grade was reclassified according to both the Fuhrman and the ISUP grading systems. Recurrence­free survival (RFS) and cancer-specific survival (CSS) were assessed through univariate and multivariate analyses. According to the Fuhrman grading system (group Fuhrman), TN and MVI were independent risk factors for postoperative recurrence in the multivariate analysis using the Cox proportional hazards model. According to the ISUP grading system (group ISUP), TN and MVI were independent risk factors for postoperative recurrence. In group Fuhrman, age, Fuhrman grade and TN were independent risk factors for CSS. In group ISUP, age, ISUP grade, and TN were independent risk factors for CSS. Furthermore, the group that was upgraded from Fuhrman grade 2 to ISUP grade 3 exhibited poorer CSS compared with the group that was reclassified from Fuhrman grade 2 to ISUP grade 2 (non-upgraded). Regardless of the nuclear grade, TN remained an independent predictor of RFS and CSS. To the best of our knowledge, this is the first report to prove the correlation between the 2012 ISUP recommendations and clinical outcomes in a Japanese ccRCC cohort. TN and upgrading to ISUP grade 3 were found to be potentially useful independent indicators of postoperative prognosis.

Pathogen screening and prognostic factors in children with severe ARDS of pulmonary origin.

BACKGROUND: Acute respiratory distress syndrome (ARDS) is one of the most lethal diseases encountered in the pediatric intensive care unit (PICU). The etiological pathogens and prognostic factors of severe ARDS of pulmonary origin in children with respiratory virus infections were prospectively investigated. METHODS: Enrolled children fulfilled the following criteria: (1) PICU admission; (2) age of 1 month to 16 years; (3) diagnosis of infectious pneumonia and respiratory virus infection; and (4) development of severe ARDS within 72 h after PICU admission. Pathogens were detected in the blood and tracheal lavage fluid using molecular techniques and a conventional culture system. The serum levels of inflammatory mediators on the day of PICU admission were examined. RESULTS: Fifty-seven patients (32 boys; median age, 9 months) were enrolled. Multiple virus infections, co-infection with bacteria/fungus, and bacteremia/fungemia were observed in 60%, 49%, and 32% of children, respectively. Adenovirus-B, measles virus, and cytomegalovirus were detected predominantly in tracheal lavage fluid. There were no statistically significant differences between non-survivors and survivors regarding the types of pathogen, incidence of multiple virus infection, gender, age, clinical features, and treatment. The serum levels of interferon (IFN)-γ and the IFN-γ/interleukin (IL)-10 ratio were higher in non-survivors. CONCLUSIONS: IFN-γ upregulation as detected on the day of PICU admission was found to be one of the possible prognostic factors affecting a fatal outcome. These results suggest that modulation of inflammatory responses is critical for the clinical management of children with ARDS.

Enhancement of bone regeneration by dual release of a macrophage recruitment agent and platelet-rich plasma from gelatin hydrogels.

Macrophages play an important role in regulating inflammatory responses and tissue regeneration. In the present study, their effect on bone remodeling is investigated by the simultaneous application of a macrophage recruiting agent, SEW2871 of a sphingosine-1 phosphate agonist, and platelet-rich plasma (PRP). The non-water soluble SEW2871 was solubilized in water through micelles formation with l-lactic acid grafted gelatin, and the resulting micelles with PRP were incorporated into gelatin hydrogels. Mixed SEW2871-micelles and PRP were released from gelatin hydrogels in a controlled fashion both in vitro and in vivo. In vitro migration assay revealed that the presence of PRP synergistically promoted SEW2871-induced macrophages migration. When applied to a bone defect of rats, the hydrogels incorporating mixed SEW2871-micelles and PRP recruited a higher number of macrophages than those hydrogels incorporating either SEW2871-micelles or PRP. The hydrogels incorporating mixed SEW2871-micelles and PRP enhanced the level of tumor necrosis factor (TNF)-α of pro-inflammatory cytokine, 3 days after application, while pro-inflammatory responses coupled with a significant increase in the expression level of osteoprotegerin (OPG) and interleukin (IL)-10 and transforming growth factor (TGF)-ß1 of anti-inflammatory cytokine were observed 10 days postoperatively. The hydrogels incorporating mixed SEW2871-micelles and PRP promoted bone regeneration to a significant great extent compared with those incorporating PBS and either SEW2871-micelles or PRP. It is concluded that macrophages recruitment contributed to PRP-induced bone regeneration.

Estimation of environmental control measures for tuberculosis transmission in care facilities for the elderly.

OBJECTIVES: Exogenous reinfections in tuberculosis (TB) have been reported among elderly patients in long-term care facilities. This study estimated the impact of upper room ultraviolet germicidal irradiation (UVGI) and negative air ionization on the reduction in TB infection assuming that current TB control failed. METHODS: Estimated probability distributions RA, RM, RUV+M, and RIO+M, denoting the reproduction numbers of cases with no intervention, wearing surgical mask of infector, upper room UVGI, and negative ionization with wearing mask, respectively were determined. RESULTS: It was assumed that 1 TB patient and 29 susceptibles stayed for 10 hours per day when ACH was 3; all subjects stayed for 60 consecutive days. The median RA increased from 7.38 (15th day) to 11.72 (two month). The percent reductions of RM, RUV+M, and RIO+M ranged from 52.4% (15th day) to 41.6% (two month), from 76.6% to 68.3%, and from 74.9% to 63.0%, respectively. The percent reductions of slopes; the change of median RM, RUV+M, and RIO+M for a change in length of stay, were estimated to be 50.8%, 87.3% and 73.7% when ACH was 1. CONCLUSIONS: In addition to case detection and source control measures, environmental control measures may be effective in preventing exogenous reinfection of TB in elderly care facilities.

Relationship between the use of an electronic commercial prescribing system and medical and medication errors in a teaching hospital.

OBJECTIVE: To investigate the effects of a commercial electronic prescribing system on medical and medication errors. METHODS: We conducted a retrospective before-and-after study in a teaching hospital by applying a U control chart to monthly error rates that were calculated from the number of both medical and medication errors divided by the number of patient days. RESULTS: The process mean medical error rate from April 2008 to December 2010 was 0.0213±0.0004 (mean±SD), and consecutive data points after September in 2010 were under the process mean from April 2008 to December 2010. The process mean value from January 2011 to March 2012 was 0.0177±0.0005. A significant difference was seen in the process mean medical error rate between consecutive months before December 2010 and values after January 2011 (p<0.001). From April 2008 to December 2010, the process mean medication error rate was 0.0060±0.0002, and from January 2011 to March 2012, the mean value was 0.0045±0.0002 (p<0.001). CONCLUSION: We showed that it requires time to reach a significant reduction in medical and medication error rates after implementation of an electronic prescribing system. It is important to check and support ways to adjust to procedures involved in using a new system.

Multivariate analysis of wound complications after surgery for laryngeal and hypopharyngeal cancers.

PURPOSE: The aim of this study was to identify the factors leading to postoperative complications following surgical management of laryngeal and hypopharyngeal cancers. PROCEDURES: Between 2001 and 2008, the medical records of 107 laryngeal and hypopharyngeal cancer patients requiring laryngectomy or pharyngolaryngectomy at our hospital were reviewed. The incidence of wound complications and correlation of complications with clinicopathological factors were investigated by univariate and multivariate analysis. RESULTS: The overall incidence of wound complication was 33.6%. The complication incidence was 35.2, 21.7 and 46.2% for the primary surgery, radiation and chemoradiation groups, respectively. Diabetes mellitus and bilateral paratracheal node dissection were significantly correlated and were independent risk factors according to multivariate analysis. Bleeding from a large vessel occurred in 4 patients, and there were significant correlations with chemoradiation. CONCLUSION: Preoperative chemoradiation was not a significant risk factor for wound complication in this study. However, once postoperative wound complications occurred, they tended to produce lethal outcomes.

Synergistic effects of the dual release of stromal cell-derived factor-1 and bone morphogenetic protein-2 from hydrogels on bone regeneration.

The objective of this study is to evaluate the activity of gelatin hydrogels incorporating combined stromal cell-derived factor-1 (SDF-1) and bone morphogenetic protein-2 (BMP-2) on the in vivo bone regeneration at an ulna critical-sized defect and subcutaneous site of rats, and compared with that of those incorporating either SDF-1 or BMP-2. The similar release profile of SDF-1 and BMP-2 from the hydrogels was observed with or without the combination of BMP-2 and SDF-1, respectively. An enhanced bone regeneration by the hydrogels incorporating combined SDF-1 and BMP-2 was observed. In addition, the implantation of hydrogels incorporating combined SDF-1 and BMP-2 enhanced the expression level of CXC chemokine cell-surface receptor-4 (Cxcr4), Runt-related factor-2 (Runx2), and Osteocalcin genes. The experiments with green fluorescent protein (GFP)-positive Chimeric mice revealed that the recruitment of bone marrow-derived cells was promoted and a vascular-like structure together with strong accumulation of CD31- and CD34-positive cells was observed at the site of hydrogels incorporating combined SDF-1 and BMP-2 implanted. In addition, a large fraction of CD29- and CD44-positive non-hematopoietic cells was detected. It is concluded that the combined release of SDF-1 and BMP-2 enhanced the recruitment of osteogenic cells and angiogenesis, resulting in the synergistic effect on bone regeneration.

Effects of liposome-encapsulated hemoglobin on human immune system: evaluation in immunodeficient mice reconstituted with human cord blood stem cells.

As preclinical evaluation in animals does not necessarily portray human responses, liposome-encapsulated hemoglobin (LEH), an artificial oxygen carrier, was tested in immunodeficient mice reconstituted with human hematopoietic stem cells (cord blood-transfused NOD/SCID/IL-2R(gammanull)[CB-NOG] mice). Changes in immunocompetent T-cell and B-cell composition in peripheral blood, spleen, and bone marrow were examined 2 and 7 days after 10 mL/kg of intravenous administration of LEH, empty liposome (EL), or saline using immunohistochemical and flow cytometrical techniques in wild-type mice and CB-NOG mice. Responses to intraperitoneal administration of toxic shock syndrome toxin-1 (TSST-1) under the absence or presence of LEH (10 mL/kg) were also determined 4 h and 3 days later in terms of lymphocyte composition and IL-2 plasma level in wild-type as well as CB-NOG mice. When liposome (LEH or EL) was administered to wild-type or CB-NOG mice, the composition of B-cells and T-cells in the spleen or peripheral blood failed to show any consistent or significant changes. The responses to a bacterial antigen (TSST-1) measured by IL-2 production were comparable regardless of the presence or absence of LEH in wild-type as well as in CB-NOG mice. Cellularity, distribution, and maturation of these human cells in peripheral blood, spleen, and bone marrow were comparable among the groups. The results suggest that simple LEH administration may not change immune cellularity, and LEH presence may not largely affect the early T-cell response to bacterial enterotoxins in murine as well as in reconstituted human immune systems.

Statistical analysis of post-operative complications after head and neck surgery.

The risk factors for wound complications after surgery for head and neck cancers at Tokai University Hospital were evaluated. The medical records of 71 head and neck cancer patients who underwent surgery between January 2000 and December 2002 were reviewed. The overall incidence of postoperative complications was 39.4%. Uni- and multivariate analyses of the risk factors leading to complications demonstrated that free flap use was significant. Further, we analyzed the relationship between radiotherapy and postoperative complications. There was no significant correlation between them.The prevention and treatment of postoperative complications were discussed.

A drink to healthy aging: The association between older women's use of alcohol and their health-related quality of life.

OBJECTIVES: To assess the relationship between alcohol intake and mortality in a cohort of women aged 70 and older and to explore the relationship between level of alcohol use and changes in physical and mental health-related quality of life. DESIGN: National longitudinal surveys from 1996 to 2002. SETTING: Community based. PARTICIPANTS: A national random sample of 12,432 Australian women aged 70 to 75 at baseline. MEASUREMENTS: Alcohol consumption was the factor of interest, and the main outcome measures were survival and health-related quality of life, with adjustment for potential confounders. RESULTS: Women who did not consume alcohol or who drank rarely were more likely to die (nondrinkers' hazard (HR) = ratio 1.94, 95% confidence interval (CI) = 1.4-2.6; rare drinkers' HR = 1.58, 95% CI = 1.2-2.1) than women in the low-intake reference category (1-2 drinks per day, 3-6 days per week), or if they survived, they had lower health-related quality-of-life scores on the General Health and Physical Functioning subscales of the Medical Outcomes Study 36-item Short Form Survey after adjustment for smoking, comorbidity, education, body mass index, and area of residence. Nondrinkers also scored lower on the Mental Health and Social Functioning subscales. CONCLUSION: Being a nondrinker of alcohol was associated with greater risk of death and poorer health-related quality of life. Results for other levels of intake were consistent with current Australian alcohol consumption recommendations for women and indicated that moderate alcohol intake may carry some health benefits for older women in terms of survival and quality of life.

Detection of individual over-smoking using conventional laboratory tests.

OBJECTIVE: The present study was designed to find useful markers for detecting the severity of smoking effects on the human body from conventional laboratory tests used in community health check-ups. METHODS: The subjects were 18,576 persons who visited Tokai University Hospital Health Check Clinic between January 5, 2000, and December 28, 2000. The data on their life-style information and laboratory tests were analyzed by multiple regression analysis and logistic analysis. RESULTS: White blood cell counts (WBC), the numbers of lymphocytes (Ly), hemoglobin concentration (Hb), mean corpuscular volume of red blood cells (MCV), and carcinoembriogenic antigen (CEA) were positively correlated with numbers of cigarettes consumed, while high density lipoprotein cholesterol (HDL-C) was negatively correlated, with statistically significant difference. Although the relationship between the grade of smoking habit and MCV, HDL-C and CEA was affected by alcohol intake, the WBC, MCV, HDL-C and CEA were shown to be the grading index for smoking effect regardless of drinking habit. Logistic analysis indicated that an accumulation of these marker abnormalities was not useful for differentiating smokers from non-smokers as compared with a single marker abnormality. CONCLUSION: WBC, MCV, HDL-C and CEA can be the grading index for smoking effect. Abnormalities in one or some of those values observed in healthy smokers indicate some effects of smoking on their bodies.