Postoperative C-reactive Protein Is a Predictive Biomarker for Survival After Non-small Cell Lung Cancer Resection.

BACKGROUND/AIM: Preoperative C-reactive protein (CRP) is well recognized as a prognostic factor of non-small cell lung cancer (NSCLC). The present study aimed to elucidate the prognostic impact of postoperative CRP in patients with NSCLC following lung resection. PATIENTS AND METHODS: We retrospectively reviewed 336 patients with NSCLC treated with lung resection. CRP levels were measured at postoperative week 6 (CRP6w; range: 4-8 weeks). Patients were divided into two groups based on CRP6w median value (5.0 mg/l); the 5-year overall survival (OS) as well as the recurrence-free survival (RFS) was evaluated in both groups. RESULTS: Five-year OS and RFS were worse in the high-CRP6w group than in the low-CRP6w group (62.9% vs. 82.9%; p<0.001, 48.4% vs. 76.1%; p<0.001, respectively). Subgroup analysis for pathological stage I and ≥II also revealed worse OS in the high-CRP6w group. Multivariate analysis revealed an association between high CRP6w and worse OS (hazard ratio, 2.23; p<0.001). CONCLUSION: CRP6w may serve as a prognostic biomarker in patients with resected NSCLC.

Impact of the favorable prognosis of patients with lung cancer adjoining bullae.

BACKGROUND: Lung cancer adjoining bullae (LC-AB) is an uncommon manifestation. The clinical characteristics and prognosis of LC-AB remain unclear. The aim of this study is to investigate the clinical features and overall survival (OS) of patients with LC-AB following lung resection compared to non-LC-AB group. METHODS: We retrospectively investigated 291 consecutive patients with lung cancer who underwent curative resection in a single institution between April 2007 and March 2015. A total of LC-AB was 52 patients. LC-AB was determined using thin slice computed tomography (CT) imaging and pathological findings. Survival analysis was calculated using the Kaplan-Meier method. We used a Cox proportional hazards model for the univariate and multivariate analysis to identify prognostic factors. RESULTS: The LC-AB group showed a higher frequency of younger patients (P=0.017), former or current smokers (P=0.011), men (P=0.021), tumor location in the upper lobe (P=0.031), moderately or poorly differentiated tumor histology (P<0.001), pleural indentation (P=0.007), and non-adenocarcinoma histology (P=0.016) than the non-LC-AB group. The 5-year survival and recurrence-free survival (RFS) rates were significantly higher in the LC-AB group than the non-LC-AB group (88.5% vs. 74.9%, P=0.010, 75.4% vs. 61.3%, P=0.030, respectively). Multivariate analysis using a Cox proportional hazard model of OS showed that LC-AB was an independent prognostic factor [hazard ratio (HR): 0.30, 95% confidence interval (CI): 0.12-0.77, P=0.012]. CONCLUSIONS: Patients with LC-AB had better OS than those with non-LC-AB. Thus, LC-AB may be an independent favorable prognostic factor following curative resection.

Impact of Postoperative Pneumonia Developing After Discharge on Long-Term Follow-up for Resected Lung Cancer.

BACKGROUND: Postoperative nosocomial pneumonia is a common immediate complication following lung resection. However, the incidence and mortality of pneumonia developing after discharge (PDAD) for lung-resected patients during long-term observation remain unclear. The aim of this study was to investigate the clinical features of PDAD in patients with resected lung cancer. METHODS: We conducted a retrospective cohort study of 357 consecutive patients with lung cancer who had undergone lung resection at a single institution, between April 2007 and December 2016. The clinical characteristics, pathological features, and overall survival were analyzed. Propensity score matched analysis was used for the evaluation of overall survival between PDAD and non-PDAD groups with adjusted relevant confounding factors. RESULTS: PDAD was observed in 66 patients (18.5%). The cumulative incidence of PDAD was 14.9% at 3 years and 21.6% at 5 years. Mortality of PDAD was 30.3%. Multivariate analysis demonstrated that the risk factors for PDAD were age (OR 1.07; P = 0.005), oral steroid use (OR 5.62; P = 0.046), and lower-lobe resection (OR 1.87; P = 0.034). After propensity score matching, 52 patients with PDAD and 52 patients without it were compared. The incidence of PDAD resulted in a worse 5-year overall survival (56.1 vs. 69.3%; P = 0.024). The Cox proportional hazards model indicated that PDAD was associated with poor overall survival (HR 1.99, P = 0.027). CONCLUSIONS: Our findings revealed a high incidence and mortality of PDAD among patients who had undergone lung resection with long-term follow-up. Therefore, PDAD could be associated with poorer overall survival.

Prognostic Impact of Postoperative C-reactive Protein for Non-small Cell Lung Cancer Following Lobectomy.

BACKGROUND/AIM: C-reactive protein (CRP) is associated with cancer progression; however, the prognostic impact of postoperative CRP remains controversial. The aim of this study was to investigate the prognostic significance of postoperative CRP in patients with resected non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: A total of 198 consecutive patients with NSCLC that had undergone lobectomy were retrospectively reviewed. CRP was measured on postoperative days 1, 3, and 5. Patients were divided into two groups according to the median of CRP on postoperative day 3 (CRP3); the high and low CRP3 groups (n=99, respectively). RESULTS: Five-year overall survival was significantly higher in the high CRP3 group than the low CRP3 (75.3% vs. 86.5%; p=0.016), as was the 5-year recurrence-free survival (62.7% vs. 73.0%; p=0.016). Multivariate analysis revealed that high CRP3 was associated with a favorable prognosis (hazard ratio(HR)=0.36; p<0.001). CONCLUSION: High CRP3 may be a favorable prognostic predictor in patients with NSCLC following lobectomy.

Healthy Aging and Concerns Among Elderly Japanese: A Text Analysis Approach.

Japan is facing an unprecedented aging society. In 2013, 25.1% of the total population in Japan was 65 years old and over. One third of the population in Japan will be at least 65 years old in 2035; healthy aging is crucial for public health and society in Japan. Under the budget limitations of social security, policies and frequent policy changes have created great uncertainty and concern for the elderly population. Given these circumstances, we conducted questionnaire surveys in 2010 to examine concerns in this age-group. In 2010, we sent questionnaires to university graduates 65 years old and older, randomly selected from a list of an alumni association of 11 universities. The questionnaires were open-ended and asked them to write any concerns that they had about health care, health policy, health systems, and their future. We received 344 responses. Among those who responded, 271 (78.8%) respondents said that they had concerns. We conducted a text analysis (Trend Search-Keyword Associator 2008) and extracted the keywords from their responses. The keywords that were extracted frequently included hospitals, doctors, health-care system, health care, Japanese government policies, government, medical insurance system for the elderly, support, increase, copayment, health (their own), scarcities, and burdens. In this study, many elderly people in Japan are concerned about health-care expenditures, health-care systems, and health policies. This may be caused by frequent health policy changes and uncertainty due to frequent administration changes such as the lack of clarity concerning raising the out of pocket payment rate from 10% to 20% for elderly people under budget constraints in Japan. Correct policy making and determination to eliminate the concerns from this vulnerable population in Japan are necessary for healthy aging.

Characterization of in vivo tumorigenicity tests using severe immunodeficient NOD/Shi-scid IL2Rγnull mice for detection of tumorigenic cellular impurities in human cell-processed therapeutic products.

The contamination of human cell-processed therapeutic products (hCTPs) with tumorigenic cells is one of the major concerns in the manufacturing and quality control of hCTPs. However, no quantitative method for detecting the tumorigenic cellular impurities is currently standardized. NOD/Shi-scid IL2Rγnull (NOG) mice have shown high xeno-engraftment potential compared with other well-known immunodeficient strains, e.g. nude mice. Hypothesizing that tumorigenicity test using NOG mice could be a sensitive and quantitative method to detect a small amount of tumorigenic cells in hCTPs, we examined tumor formation after subcutaneous transplantation of HeLa cells, as a model of tumorigenic cells, in NOG mice and nude mice. Sixteen weeks after inoculation, the 50% tumor-producing dose (TPD50) values of HeLa cells were stable at 1.3 × 104 and 4.0 × 105 cells in NOG and nude mice, respectively, indicating a 30-fold higher sensitivity of NOG mice compared to that of nude mice. Transplanting HeLa cells embedded with Matrigel in NOG mice further decreased the TPD50 value to 7.9 × 10 cells, leading to a 5000-fold higher sensitivity, compared with that of nude mice. Additionally, when HeLa cells were mixed with 106 or 107 human mesenchymal stem cells as well as Matrigel, the TPD50 values in NOG mice were comparable to those of HeLa cells alone with Matrigel. These results suggest that the in vivo tumorigenicity test using NOG mice with Matrigel is a highly sensitive and quantitative method to detect a trace amount of tumorigenic cellular impurities in human somatic cells, which can be useful in the quality assessment of hCTPs.

[Psychophysiological effects of hand massage in geriatric facility residents].

OBJECTIVES: Geriatric facility residents experience easy decrease of their quality of life (QOL) compared with elderly people cared for in their own homes owing to environmental changes. Because complementary and alternative medicine is useful to improve QOL, a simple manual for hand massage for inexperienced volunteers was developed, and the physical and psychological effects of this method were examined. METHODS: Thirty-six female geriatric facility residents (mean age, 84.5 years) were chosen. They were divided into two groups: the control and hand-massaged groups. In the hand-massaged group, females received 15 min hand massage from volunteers in accordance with a simple manual. No hand massage was performed in the control group. Before and after the treatment, a survey using a questionnaire (7 items; e.g., tension, fatigue) and physiological measurements (e.g., blood pressure, hand skin temperature), and measurement of cortisol and chromogranin A levels in saliva samples using enzyme-linked immunosorbent assay kits were carried out. Data were analyzed by two-way analysis of variance (ANOVA) to determine the effects of group and time on the dependent variables. RESULTS: The hand-massaged group showed greater increases in skin temperature than the control group, as shown by ANOVA (p<.05). Results showed that improvements of the scores in the questionnaire (condition and feel-good items) were greater in the hand-massaged group than in the control group (p<.05). Cortisol level increased in the control group, but was not statistically significantly different from that in the hand-massaged group, as shown by ANOVA. Chromogranin A level was also not significantly different between the two groups. CONCLUSIONS: These results confirmed the benefits of complementary and alternative medicine including touching and massage found in previous studies. Hand massage prompted relaxation in geriatric facility residents in this experiment.

Effects of psychological stress on neutrophil phagocytosis and bactericidal activity in humans--a meta-analysis.

Several authors have reviewed the effects of psychological stress on lymphocyte activity. However the effect of psychological stress on neutrophil functions has not been reviewed. The present meta-analysis summarizes evidence of the effects of psychological stress on neutrophil phagocytosis and bactericidal activity collated from a MEDLINE search of the English literature. We searched the database to identify the relevant studies through April 30, 2013. Eleven studies met our inclusion criteria and we divided them into those addressing transient acute stress (3 studies, n=74), academic examinations (4 studies n=101) and chronic stress/life events (4 studies, n=193). We performed a meta-analysis of the data and calculated total standardized mean differences (SMD) to evaluate the effects of chronic stress. Transient acute stressors might both enhance and decrease these neutrophil functions. Academic examinations tended to elevate neutrophil functions. On the other hand, the total SMDs of neutrophil phagocytosis and bactericidal activity altered by chronic stress/life events were -0.589 (95% CI: -0.908 to -0.270, p<0.05) and -0.547 (95% CI: -0.845 to -0.248, p<0.05), respectively, indicating suppressive effects on these neutrophil functions. Further systematic review of more pooled studies is warranted to confirm that academic examinations might enhance, whereas chronic stress/life events might suppress these neutrophil functions.

Carcinogenicity evaluation for the application of carbon nanotubes as biomaterials in rasH2 mice.

The application of carbon nanotubes (CNTs) as biomaterials is of wide interest, and studies examining their application in medicine have had considerable significance. Biological safety is the most important factor when considering the clinical application of CNTs as biomaterials, and various toxicity evaluations are required. Among these evaluations, carcinogenicity should be examined with the highest priority; however, no report using transgenic mice to evaluate the carcinogenicity of CNTs has been published to date. Here, we performed a carcinogenicity test by implanting multi-walled CNTs (MWCNTs) into the subcutaneous tissue of rasH2 mice, using the carbon black present in black tattoo ink as a reference material for safety. The rasH2 mice did not develop neoplasms after being injected with MWCNTs; instead, MWCNTs showed lower carcinogenicity than carbon black. Such evaluations should facilitate the clinical application and development of CNTs for use in important medical fields.

Effects of life events and stress on neutrophil functions in elderly men.

BACKGROUND: Psychological stress or life events might significantly decrease neutrophil function in elderly individuals and lead to infectious diseases. However, relationships between these factors have not been examined in detail.We investigated the relationships between neutrophil function measured using the nitroblue tetrazolium (NBT) reduction test and measurements of psychological stress and life events among 81 men aged over 60 years. RESULTS: The numbers and scores for life events were significantly higher (p < 0.01, respectively) in a group with values reflecting phagocytosis below the median than in a control group. CONCLUSIONS: Chronic psychological stress due to life events decreases neutrophil functions among elderly men.

[Changes in mental health index, serum biochemical parameters and neutrophil function after start of the wheelchair use at a nursing home in Korea].

OBJECTIVES: In Korea, long-term care insurance has been provided to the elderly since 2008. We conducted a health survey of the elderly in a nursing home in Korea from 2007 to 2010. Since the introduction of the insurance system, many of the elderly have started using wheelchairs. We aimed to determine the effect of using wheelchairs on mental health index, serum biochemical parameters, and neutrophil function. METHODS: The subjects were 32 elderly persons (mean age: 74.44 ± 5.79 years) who could walk unaided in 2007 and were not bedridden until 2010. We classified the subjects into the walking group (subjects could walk unaided or with a cane until 2010; 10 men, 9 women) and the wheelchair group (subjects required the use of a wheelchair from 2007 to 2010; 2 men, 11 women). We determined stress score, stress tolerance, subjective well-being score in accordance with the Philadelphia Geriatric Center Morale Scale, and serum biochemical parameters. We also analyzed oxidant production and phagocytosis by neutrophils and estimated the balance between oxidant production and phagocytosis by correlation analysis. RESULTS: Increases in the stress tolerance of the wheelchair group in 2008 and 2009 and the subjective well-being score in 2009 in the survey were observed. However, in 2010, the stress tolerance and the subjective well-being score decreased to the levels in 2007. In 2010, the levels of serum albumin and high-density lipoprotein cholesterol of the wheelchair group were significantly lower than those of the walking group, and the balance between phagocytosis and oxidant production in the wheelchair group improved.

Detection of the onset of ischemia and carcinogenesis by hypoxia-inducible transcription factor-based in vivo bioluminescence imaging.

An animal model for the early detection of common fatal diseases such as ischemic diseases and cancer is desirable for the development of new drugs and treatment strategies. Hypoxia-inducible factor 1 (HIF-1) is a transcription factor that regulates oxygen homeostasis and plays key roles in a number of diseases, including cancer. Here, we established transgenic (Tg) mice that carry HRE/ODD-luciferase (HOL) gene, which generates bioluminescence in an HIF-1-dependent manner and was successfully used in this study to monitor HIF-1 activity in ischemic tissues. To monitor carcinogenesis in vivo, we mated HOL mice with rasH2 Tg mice, which are highly sensitive to carcinogens and are used for short-term carcinogenicity assessments. After rasH2-HOL Tg mice were treated with N-methyl-N-nitrosourea, bioluminescence was detected noninvasively as early as 9 weeks in tissues that contained papillomas and malignant lesions. These results suggest that the Tg mouse lines we established hold significant potential for monitoring the early onset of both ischemia and carcinogenesis and that these lines will be useful for screening chemicals for carcinogenic potential.

[Effect of lifestyle and health behavior on neutrophil function].

OBJECTIVE: In this study, we investigated the relationships of neutrophil functions with lifestyle factors (namely, subjective stress, exercise habits, smoking habits, alcohol-drinking habits, and self-perceived status health) and health behavior in the Chinese urban elderly. METHODS: We performed a health survey of the elderly aged 65 years or older living in Tianjin. The subjects were 42 males (69.1 ± 4.1 years old) and 41 females (69.1 ± 4.1 years old). Investigations of subjective stress, exercise habits, smoking habits, alcohol-drinking habits, and self-perceived health status were performed. The phagocytosis and superoxide productivity of neutrophils were measured by the nitroblue tetrazolium (NBT) reduction method. In addition, leukocyte count and serum total protein (TP) level were examined. RESULTS: The investigations revealed the associations of health behavior (p<0.05) and self-perceived health (p<0.10) with the balance between phagocytosis and subsequent superoxide production. CONCLUSIONS: The present study revealed that there were correlations of neutrophil functions with lifestyle factors (subjective stress, exercise habits, smoking habits, alcohol-drinking habits, and self-perceived status health) and health behavior in the Chinese urban elderly.

Casein kinase 1 delta regulates the pace of the mammalian circadian clock.

Both casein kinase 1 delta (CK1delta) and epsilon (CK1epsilon) phosphorylate core clock proteins of the mammalian circadian oscillator. To assess the roles of CK1delta and CK1epsilon in the circadian clock mechanism, we generated mice in which the genes encoding these proteins (Csnk1d and Csnk1e, respectively) could be disrupted using the Cre-loxP system. Cre-mediated excision of the floxed exon 2 from Csnk1d led to in-frame splicing and production of a deletion mutant protein (CK1delta(Delta2)). This product is nonfunctional. Mice homozygous for the allele lacking exon 2 die in the perinatal period, so we generated mice with liver-specific disruption of CK1delta. In livers from these mice, daytime levels of nuclear PER proteins, and PER-CRY-CLOCK complexes were elevated. In vitro, the half-life of PER2 was increased by approximately 20%, and the period of PER2::luciferase bioluminescence rhythms was 2 h longer than in controls. Fibroblast cultures from CK1delta-deficient embryos also had long-period rhythms. In contrast, disruption of the gene encoding CK1epsilon did not alter these circadian endpoints. These results reveal important functional differences between CK1delta and CK1epsilon: CK1delta plays an unexpectedly important role in maintaining the 24-h circadian cycle length.

Higher susceptibility of NOG mice to xenotransplanted tumors.

The purpose of tumorigenicity testing, as applied not only to cell substrates used for viral vaccine manufacture but also stem cells used for cell-based therapy, is to discriminate between cells that have the capacity to form tumors and cells that do not. Therefore, tumorigenicity testing is essential in assessing the safety of these biological materials. Recently developed NOD/Shi-scid IL2Rg(null) (NOG) mice have been shown to be superior to NOD/Shi-scid (SCID) mice for xenotransplantation of both normal and cancerous cells. To select a suitable mouse strain as a xenogenic host for tumorigenicity testing, we compared the susceptibility of NOG (T, B, and NK cell-defective), SCID (T and B cell-defective), and the traditionally used nude (T cell-defective) mice to tumor formation from xenotransplanted HeLa S3 cells. When 10(4) HeLa S3 cells were subcutaneously inoculated into the flanks of these mice, the tumor incidence on day 22 was 10/10 (100%) in NOG, 2/10 (20%) in SCID, and 0/10 (0%) in nude mice. The subcutaneous tumors formed reproducibly and semiquantitatively in a dose-dependent manner. Unexpectedly, half of the NOG mice (5/10) that had been inoculated with a mere 10(1) HeLa S3 cells formed progressively growing subcutaneous tumors on day 78. We confirmed that the engrafted tumors originated from inoculated HeLa S3 cells by immunohistochemical staining with anti-HLA antibodies. These data suggest that NOG mice may be the best choice as a suitable strain for testing tumorigenicity.

Calcitonin gene-related peptide stimulates proliferation of alveolar epithelial cells.

BACKGROUND: Alveolar epithelial cells are known as progenitor cells for the restoration from the damage in the lung. Calcitonin gene-related peptide (CGRP) has been reported to play an important role in the proliferation of various types of epithelial and endothelial cells. We investigated the effects of CGRP on the proliferation of alveolar epithelial cells in vitro and in vivo. METHODS: A549 cells were cultured in Dulbecco Modified Eagle Medium with 5% fatal bovin serum for 24 hours, then CGRP was added in vitro. The proliferation of DNA synthesis was measured using 5-bromo-2-deoxyuridine, an analog of thymidine, by enzyme-linked immunosorbent assay.As one intracellular response to CGRP, we examined activation of p44/42- extracellular signal-regulated kinase (ERK) pathway by adding CGRP, using western blotting method.Recombinant adenovirus encoding nuclear-targeted-human beta-CGRP (rhCGRP) was administered into Male Wister rat (n = 5, 10 weeks old) lungs by intratracheal instillation in vivo. 7 days after the administration of CGRP, rat lungs were harvested and histological findings and immunohistochemical staining of proliferating cell nuclear antigen (PCNA) were evaluated to examine cell proliferation. RESULTS: In vitro study, CGRP increased the proliferation of A549 cells in a dose and time dependent manner. CGRP8-37 (inhibitor of CGRP receptor) decreased CGRP induced proliferation of DNA synthesis. Phosphorylation of ERK pathway was observed within 15 minutes and peaked in one hour. U0126 (inhibitor of ERK pathway) decreased CGRP induced proliferation of DNA synthesis.In vivo study, histological examination of the lung indicated proliferation of alveolar epithelial cells in the rhCGRP-treated group and the nuclei of alveolar epithelial cells were positive for PCNA immunostaining. CONCLUSION: In this study, we conclude that CGRP stimulates proliferation of human alveolar epithelial cells in vivo and in vitro.

Carcinogenic comparative study on rasH2 mice produced by two breeding facilities.

CByB6F1-Tg(HRAS)2Jic mice (brand name: rasH2 mouse) are produced by two breeding facilities, CLEA Japan, Inc. (Fuji, Shizuoka, Japan) and Taconic (Germantown, NY, USA), and supplied world wide. To confirm carcinogenic conformity of both mice, a 26-week carcinogenicity test was performed on a total of 120 mice obtained from both facilities under the same protocol and same timing in our facility. All mice were divided into a vehicle (citrate buffer at pH 4.5, 10 ml/kg, single intraperitoneal injection) group and a MNU (N-methyl-N-nitrosourea, 75 mg/kg, single intraperitoneal injection) group. Fifteen mice of each sex were assigned to each group. The survival rate of the vehicle group was maintained at 100% for mice from both facilities at completion of the test. In the MNU group, MNU-induced tumor death occurred from 9 to 12 weeks after administration, and the final survival rate for both facilities was 6.7%. In the pathological examination, only benign tumors of lungs, spleen, forestomach and skin were observed in a few mice in the vehicle group of both facilities. In the MNU group, the incidence of forestomach papilloma/squamous cell carcinoma in mice from both facilities was 100%. The incidences of malignant lymphoma in CLEA Japan mice and Taconic mice were 86.7% and 93.3%, respectively, and no significant difference was observed (Fisher's exact probability test). Although lung adenoma and skin papilloma/keratoacanthoma, which are major MNU induced tumors in this strain, were observed in several mice from both facilities, no significant differences were found. Consequently, carcinogenic conformity of rasH2 mice derived from two breeding facilities was confirmed by the present study.

Examination of percutaneous application in a 26-week carcinogenicity test in CB6F1-TG rasH2 mice.

We examined the possibility of expanding applications of rasH2 mice, which are genetically manipulated mice for short-term carcinogenicity tests, to percutaneous application. A 26-week short-term carcinogenicity study was performed on a total of 300 mice including 75 male and female rasH2 mice each, and 75 male and female non-Tg mice each from the same litter as the rasH2 mice divided into untreated group, an ethanol group, a white Vaseline group, an acetone group, and a phorbol 12-myristate 13-acetate (TPA) group. Only shaving of dorsal skin was performed on the untreated mice. As a positive control, TPA was administered percutaneously at a dose of 2.5 microg/kg and 3 times/week for 26 weeks based on the protocol for Tg.AC mice in the ILSI/HESI international validation study. In the ethanol, white Vaseline, and acetone groups, no tumorous changes were observed on the skin at the administration site. In the TPA group, nodular changes at the administration site were observed from seven weeks after the start of administration in rasH2 mice, and the incidence in males and females was 50.0% (7/14) and 53.3% (8/15), respectively. In a pathological examination, nodules in 21.4% (3/14) of males and 46.7% (7/15) of females were diagnosed as skin papilloma or keratoacanthoma, and the rest as squamous cell hyperplasia. In the non-Tg mice, no nodules or tumorigenic changes were observed at the administration site. These findings show that percutaneous application in rasH2 mice is possible in 26-week carcinogenicity tests.