[Retracted] Mst1 regulates non­small cell lung cancer A549 cell apoptosis by inducing mitochondrial damage via ROCK1/F­actin pathways.

Subsequently to the publication of the above article, an interested reader drew to the authors' attention that certain of the EdU assay data shown in Fig. 7E on p. 2418 had already appeared in different form in a previously published paper written by different authors at different research institutes. Owing to the fact that the contentious data in the above article had already been published prior to its submission to International Journal of Oncology, the Editor has decided that this paper should be retracted from the Journal. The authors were asked for an explanation to account for these concerns, but the Editorial Office did not receive a reply. The Editor apologizes to the readership for any inconvenience caused.  [International Journal of Oncology 53: 2409­2422, 2018; DOI: 10.3892/ijo.2018.4586].

Design, synthesis and biological evaluation of novel DCLK1 inhibitor containing purine skeleton for the treatment of pancreatic cancer.

Pancreatic cancer is a highly lethal form of malignancy that continues to pose a significant and unresolved health challenge. Doublecortin-like kinase 1 (DCLK1), a serine/threonine kinase, is found to be overexpressed in pancreatic cancer and holds promise as a potential therapeutic target for this disease. However, few potent inhibitors have been reported currently. Herein, a series of novel purine, pyrrolo [2,3-d]pyrimidine, and pyrazolo [3,4-d] pyrimidine derivatives were designed, synthesized, and evaluated their biological activities in vitro. Among them, compound I-5 stood out as the most potent compound with strong inhibitory activity against DCLK1 (IC50 = 171.3 nM) and remarkable antiproliferative effects on SW1990 cell lines (IC50 = 0.6 µM). Notably, I-5 exhibited higher in vivo antitumor potency (Tumor growth inhibition value (TGI): 68.6 %) than DCLK1-IN-1 (TGI: 24.82 %) in the SW1990 xenograft model. The preliminary mechanism study demonstrated that I-5 not only inhibited SW1990 cell invasion and migration, but also decreased the expression of prominin-1 (CD133) and cluster of differentiation 44 (CD44), which are considered as differentiation markers for SW1990 stem cells. All the results indicated that I-5, a novel DCLK1 inhibitor, shows promise for further investigation in the treatment of pancreatic cancer.

An outbreak of pulmonary tuberculosis and a follow-up investigation of latent tuberculosis in a high school in an eastern city in China, 2016-2019.

BACKGROUND: In October 2016, a senior high school student was diagnosed with sputum-smear positive [SS(+)] pulmonary tuberculosis (TB). We conducted an investigation of an outbreak in the school, including among students and teachers diagnosed with latent TB, who we followed until July 2019. METHODS: We defined latent TB infection (LTBI) as a tuberculin skin test (TST) induration of 15mm or larger; probable TB as a chest radiograph indicative of TB plus productive cough/hemoptysis for at least 2 weeks, or TST induration of 15mm or larger; and confirmed TB as two or more positive sputum smears or one positive sputum smear plus a chest radiograph indicative of TB or culture positive with M. tuberculosis. We conducted mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) typing based on 24 loci in the isolates. RESULTS: Between October 2016 and July 2019, we identified 52 cases, including nine probable, six confirmed, and 37 LTBI cases. The index case-student had attended school continuously despite having TB symptoms for almost three months before being diagnosed with TB. We obtained three isolates from classmates of the index case in 2016; all had identical MIRU-VNTR alleles with the index case. The LTBI rate was lower among students (7.41%, 30/405) than among teachers (26.92%, 7/26) (rate ratio [RR] = 0.28, 95% confidential interval [CI]: 0.13-0.57). Among the 17 students who had latent TB and refused prophylaxis in October 2016, 23.53% (4/17) became probable/confirmed cases by July 2019. None of the six teachers who also refused prophylaxis became probable or confirmed cases. Of the 176 students who were TST(-) in October 2016, 1.70% (3/176) became probable/confirmed cases, and among the 20 teachers who were TST(-), 1 became a probable case. CONCLUSIONS: Delayed diagnosis of TB in the index patient may have contributed to the start of this outbreak; lack of post-exposure chemoprophylaxis facilitated spread of the outbreak. Post-exposure prophylaxis is strongly recommended for all TST-positive students; TST-negative students exposed to an SS(+) case should be followed up regularly so that prophylaxis can be started if LTBI is detected.

Simulation exercises and after action reviews - analysis of outputs during 2016-2019 to strengthen global health emergency preparedness and response.

BACKGROUND: Under the International Health Regulations (2005) [IHR (2005)] Monitoring and Evaluation Framework, after action reviews (AAR) and simulation exercises (SimEx) are two critical components which measure the functionality of a country's health emergency preparedness and response under a "real-life" event or simulated situation. The objective of this study was to describe the AAR and SimEx supported by the World Health Organization (WHO) globally in 2016-2019. METHODS: In 2016-2019, WHO supported 63 AAR and 117 SimEx, of which 42 (66.7%) AAR reports and 56 (47.9%) SimEx reports were available. We extracted key information from these reports and created two central databases for AAR and SimEx, respectively. We conducted descriptive analysis and linked the findings according to the 13 IHR (2005) core capacities. RESULTS: Among the 42 AAR and 56 SimEx available reports, AAR and SimEx were most commonly conducted in the WHO African Region (AAR: n = 32, 76.2%; SimEx: n = 32, 52.5%). The most common public health events reviewed or tested in AAR and SimEx, respectively, were epidemics and pandemics (AAR: n = 38, 90.5%; SimEx: n = 46, 82.1%). For AAR, 10 (76.9%) of the 13 IHR core capacities were reviewed at least once, with no AAR conducted for food safety, chemical events, and radiation emergencies, among the reports available. For SimEx, all 13 (100.0%) IHR capacities were tested at least once. For AAR, the most commonly reviewed IHR core capacities were health services provision (n = 41, 97.6%), risk communication (n = 39, 92.9%), national health emergency framework (n = 39, 92.9%), surveillance (n = 37, 88.1%) and laboratory (n = 35, 83.3%). For SimEx, the most commonly tested IHR core capacity were national health emergency framework (n = 56, 91.1%), followed by risk communication (n = 48, 85.7%), IHR coordination and national IHR focal point functions (n = 45, 80.4%), surveillance (n = 31, 55.4%), and health service provision (n = 29, 51.8%). For AAR, the median timeframe between the end of the event and AAR was 125 days (range = 25-399 days). CONCLUSIONS: WHO has recently published guidance for the planning, execution, and follow-up of AAR and SimEx. Through the guidance and the simplified reporting format provided, we hope to see more countries conduct AAR and SimEx and standardization in their methodology, practice, reporting and follow-up.

Long Noncoding RNA HIF1A-AS2 Promotes Non-Small Cell Lung Cancer Progression by the miR-153-5p/S100A14 Axis.

BACKGROUND: Long noncoding RNA (lncRNA) plays a critical role in initiating lung cancer. This study aims to research the function and mechanism of lncRNA HIF1A-AS2 in regulating non-small cell lung cancer (NSCLC) progression. METHODS: qRT-PCR was used to analyze gene expression. The CCK-8 assay was performed to detect cell proliferation. The Transwell assay was conducted to examine cell migration and invasion. A Caspase3 activity detection kit was utilized to analyze apoptosis. The luciferase reporter assay was carried out to research interactions of HIF1A-AS2, miR-153-5p and S100A14. RESULTS: HIF1A-AS2 expression was raised in NSCLC tissues and cell lines. The HIF1A-AS2 level was increased in advanced NSCLC tumor tissues. High HIF1A-AS2 expression was related to poor prognosis. HIF1A-AS2 knockdown decreased proliferation, migration and invasion while promoting apoptosis. HIF1A-AS2 was the sponge for miR-153-5p, and miR-153-5p targeted S100A14. HIF1A-AS2 promoted S100A14 expression through regulating miR-153-5p. CONCLUSION: The HIF1A-AS2/miR-153-5p/S100A14 axis plays a crucial role in promoting NSCLC progression.

Histone H2A Ubiquitination Reinforces Mechanical Stability and Asymmetry at the Single-Nucleosome Level.

Monoubiquitination at lysine 119 of histone H2A (ubH2A) is a prevalent post-translational modification that is associated with gene repression in the context of chromatin. However, the direct function of ubH2A on nucleosome is poorly understood. Here we identified the effect of ubH2A on nucleosome using single-molecule magnetic tweezers. We revealed that ubH2A stabilizes the nucleosome by blocking the peeling of DNA from the histone octamer. Each ubH2A reinforces one-half of the outer wrap and introduces a robust asymmetry for nucleosome unfolding. Furthermore, a real-time deubiquitination process confirmed that ubH2A-nucleosome is sequentially deubiquitinated and restored to the unmodified nucleosome state. These results provide a novel mechanism to understand the repression of the passage of RNA or DNA polymerases through the ubH2A-nucleosome barrier during gene transcription or replication.

Association between PM2.5 Exposure and All-Cause, Non-Accidental, Accidental, Different Respiratory Diseases, Sex and Age Mortality in Shenzhen, China.

Background: China is at its most important stage of air pollution control. Research on the association between air pollutants and human health is very important and necessary. The purpose of this study was to evaluate the association between PM2.5 concentrations and residents' mortality and to compare the effect of PM2.5 on the different diseases, accidental deaths, sex or age of residents from high polluted areas with less polluted areas. Methods: The semi-parametric generalized additive model (GAM) with Poisson distribution of time series analysis was used. The excess risk (ER) of mortality with the incremental increase of 10 µg/m³ in PM2.5 concentration was calculated. Concentration-response relationship curves and autocorrelation between different lags of PM2.5 were also evaluated. Results: PM2.5 exposure was significantly associated with the mortality of residents. The strongest ERs per 10 µg/m³ increase in PM2.5 were 0.74% (95% CI: 0.11⁻1.38%) for all-cause, 0.67% (95% CI: 0.01⁻1.33%) for non-accidental, 1.81% (95% CI: 0.22⁻3.42%) for accidental, 3.04% (95% CI: 0.60⁻5.55%) for total respiratory disease, 6.38% (95% CI: 2.78⁻10.11%) for chronic lower respiratory disease (CLRD), 8.24% (95% CI: 3.53⁻13.17%) for chronic obstructive pulmonary disease (COPD), 1.04% (95% CI: 0.25⁻1.84%) for male and 1.32% (95% CI: 0.46⁻2.19%) for elderly. Furthermore, important information on the concentration-response relationship curves was provided. Conclusions: PM2.5 can increase the risk of residents' mortality, even in places with less air pollution and developed economy in China.

Mst1 regulates non-small cell lung cancer A549 cell apoptosis by inducing mitochondrial damage via ROCK1/F­actin pathways.

Mammalian STE20-like kinase 1 (Mst1) is well recognized as a major tumor suppressor in cancer development, growth, metabolic reprogramming, metastasis, cell death and recurrence. However, the roles of Mst1 in non-small cell lung cancer (NSCLC) A549 cell phenotypic alterations remain to be elucidated. The present study aimed to explore the functional role and underlying mechanisms of Mst1 with regards to A549 cell proliferation, migration and apoptosis; this study focused on mitochondrial homeostasis and Rho-associated coiled-coil containing protein kinase 1 (ROCK1)/F­actin pathways. The results demonstrated that Mst1 was downregulated in A549 cells compared with in a normal pulmonary epithelial cell line. Subsequently, overexpression of Mst1 in A549 cells reduced cell viability and promoted cell apoptosis. Furthermore, overexpression of Mst1 suppressed A549 cell proliferation and migration. At the molecular level, the reintroduction of Mst1 in A549 cells led to activation of mitochondrial apoptosis, as evidenced by a reduction in mitochondrial potential, overproduction of ROS, cytochrome c release from the mitochondria into the nucleus, and upregulation of pro-apoptotic protein expression. In addition, Mst1 overexpression was closely associated with impaired mitochondrial respiratory function and suppressed cellular energy metabolism. Functional studies illustrated that Mst1 overexpression activated ROCK1/F-actin pathways, which highly regulate mitochondrial function. Inhibition of ROCK1/F-actin pathways in A549 cells sustained mitochondrial homeostasis, alleviated caspase-9-dependent mitochondrial apoptosis, enhanced cancer cell migration and increased cell proliferation. In conclusion, these data firmly established the regulatory role of Mst1 in NSCLC A549 cell survival via the modulation of ROCK1/F-actin pathways, which may provide opportunities for novel treatment modalities in clinical practice.

Thoracic outlet syndrome (TOS): A case report of a rare complication after Nuss procedure for pectus excavatum.

RATIONALE: The Nuss procedure has become a major alternative operation for patients with pectus excavatum (PE). PATIENT CONCERNS: We report a case of 27-year-old man with PE who developed thoracic outlet syndrome (TOS) after the Nuss procedure. The patient showed clinical symptoms of brachial plexus compression. DIAGNOSES: Further evaluation demonstrated a narrowed space between the first rib and the anterior scalene muscle and compressing the brachial plexus and vessels. INTERVENTIONS: Nerve nourishing medicine and rehabilitation exercising were taken to restore the muscle strength. OUTCOMES: Several months later, the clinical symptoms disappeared. LESSONS: Medicine and rehabilitation exercising may benefit the functional recovery of impaired nerve in TOS in the early stage of TOS.

Association between Airborne Fine Particulate Matter and Residents' Cardiovascular Diseases, Ischemic Heart Disease and Cerebral Vascular Disease Mortality in Areas with Lighter Air Pollution in China.

BACKGROUND: China began to carry out fine particulate matter (PM2.5) monitoring in 2013 and the amount of related research is low, especially in areas with lighter air pollution. This study aims to explore the association between PM2.5 and cardiovascular disease (CVD), ischemic heart disease (IHD) and cerebral vascular disease (EVD) mortality in areas with lighter air pollution. METHODS: Data on resident mortality, air pollution and meteorology in Shenzhen during 2013⁻2015 were collected and analyzed using semi-parametric generalized additive models (GAM) with Poisson distribution of time series analysis. RESULTS: Six pollutants were measured at seven air quality monitoring sites, including PM2.5, PM10, SO2, NO2, CO and O3. The PM2.5 daily average concentration was 35.0 ± 21.9 µg/m³; the daily average concentration range was from 7.1 µg/m³ to 137.1 µg/m³. PM2.5 concentration had significant effects on CVD, IHD and EVD mortality. While PM2.5 concentration of lag5 and lag02 rose by 10 µg/m³, the excess risk (ER) of CVD mortality were 1.50% (95% CI: 0.51⁻2.50%) and 2.09% (95% CI: 0.79⁻3.41%), respectively. While PM2.5 concentration of lag2 and lag02 rose by 10 µg/m³, the ER of IHD mortality were 2.87% (95% CI: 0.71⁻5.07%) and 3.86% (95% CI: 1.17⁻6.63%), respectively. While PM2.5 concentration of lag4 and lag04 rose by 10 µg/m³, the ER of EVD mortality were 2.09% (95% CI: 2.28⁻3.92%) and 3.08% (95% CI: 0.68⁻5.53%), respectively. CONCLUSIONS: PM2.5 increased CVD mortality. The government needs to strengthen the governance of air pollution in areas with a slight pollution.

Video-assisted thoracoscopic surgery for non-small-cell lung cancer in elderly patients: a single-center, case-matched study.

The incidence of non-small-cell lung cancer among elderly patients has increased; therefore, older patients are increasingly being considered for radical pulmonary resection. However, data regarding the outcome of video-assisted thoracoscopic surgery (VATS) in elderly patients are limited. The aim of this study was to evaluate the safety and feasibility of VATS in elderly patients with non-small-cell lung cancer. From January 2008 to January 2014, a total of 78 patients aged ≥ 70 years (elderly group) undergoing VATS for NSCLC were matched with 78 patients < 70 years (young group) by demographics, tumor characteristics, and details of surgical procedures. The elderly group was characterized by a higher incidence of hypertension (P = 0.001) and diabetes mellitus (P = 0.014), and ≥ 2 comorbidities (P = 0.009). Intraoperative variables, such as surgical duration blood loss, and transfusion rate, were not notably different between the groups. Postoperative 30-day mortality, 30-day complications, and 30-day major complications were comparable between the groups. The 5-year overall survival rates were 69% in the young group and 64% in the elderly group, respectively (P = 0.258). The 5-year disease-free survival rates were 65% in the young group and 60% in the elderly group, respectively (P = 0.327). Our results clearly demonstrated that VATS for non-small-cell lung cancer could be safely and efficacy performed in elderly patients; thus, advanced age itself should not be regarded as a contraindication for VATS.

A patient with a large intrathoracic malignant schwannoma who showed a complete clinical response to rAd-p53-combined with radiotherapy.

The prognosis of postoperatively recurred malignant schwannoma is poor and there is no effective treatment. We had a patient who was found to have a large intrathoracic tumor 1 year after surgery and could not tolerate an operation for the second time. We then decided to evaluate the synergistic effect of recombinant adenovirus-p53 (rAd-p53) combined with radiotherapy for the patient. rAd-p53 was injected intratumorally twice a week before radiotherapy, a total of 10 times, over a course of treatment. Radiotherapy then followed gene therapy at five fractions a week for 5 weeks, with a total dosage of 80.6 Gy/31f in the center part of the tumor and 62 Gy/31f in other locations. The pathological diagnosis of malignant schwannoma indicated that the p53 expression was strongly positive and vascular endothelial growth factor and Bcl-2 were positive before treatment on protein immunohistochemical staining. After treatment, the diameter of the tumor was noticeably reduced and the center part of the tumor presented as a fluid anechoic area and cavities on computed tomographic scanning. The result of the puncture biopsy showed that there were many fibronecrotic tissues and no significant tumor cells. The p53 expression was weakly positive, Vascular endothelial growth factor was negative, and Bcl-2 was weakly positive after treatment on protein immunohistochemical staining.

[Effectiveness of minimally invasive surgery for pectus excavatum in adults].

OBJECTIVE: To investigate the effect of anterior chest wall depression on the cardiac function and the effectiveness of minimally invasive surgery for pectus excavatum by comparing cardiac function and morphology between pre- and post-operation. METHODS: Between August 2009 and December 2010, 102 adult patients with pectus excavatum were treated with minimally invasive surgery, including the primary operation in 95 cases and the reoperation in 7 cases. There were 84 males and 18 females, aged 18-57 years (mean, 23.4 years). The haller index (HI) was 4.59 +/- 1.51. Of 102 patients, 59 were classified as pectus excavatum type I and 43 as type II; 42 had clinical symptoms and 19 had the physical sign of heart. The preoperative chest CT examination showed cardiac compression in all patients and heart displacement in 74 patients. The left ventricular ejection fraction (LVEF) was 68.9% +/- 6.2%. RESULTS: The procedure was successful in all patients, and no death or serious complication occurred. The patients were followed up 12-28 months (mean, 21 months). The clinical symptoms and cardiac physical sign of the patients disappeared after operation. HI was 2.70 +/- 0.33 at 12 months after operation, showing significant difference when compared with preoperative HI (t = 5.83, P = 0.00). According to Nuss's evaluation method, the results were excellent in 99 patients and good in 3 patients. CT examination showed complete relief of cardiac compression in 101 patients and mild cardiac compression in 1 patient; the heart position was normal at 12 months after operation. Electrocardiogram returned to normal in 4 patients having abnormal electrocardiogram. LVEF was 70.5% + 4.8% after operation, showing no significant difference when compared with preoperative LVEF (t = 1.08, P = 0.30). CONCLUSION; The main effects of pectus excavatum in adults on heart are compression and displacement. Cardiac compression may be relieved efficiently and the patient's clinical symptoms can be abated by minimally invasive surgery.

New system for multilocus variable-number tandem-repeat analysis of the enterohemorrhagic Escherichia coli strains belonging to three major serogroups: O157, O26, and O111.

Enterohemorrhagic Escherichia coli (EHEC), a food- and waterborne pathogen, causes diarrhea, hemorrhagic colitis, and life-threatening HUS. MLVA is a newly developed and widely accepted genotyping tool. An MLVA system for EHEC O157 involving nine genomic loci has already been established. However, the present study revealed that the above-mentioned MLVA system cannot analyze EHEC O26 and O111 isolates-the second and third most dominant EHEC serogroups in Japan, respectively. Therefore, with several modifications to the O157 system and the use of nine additional loci, we developed an expanded MLVA system applicable to EHEC O26, O111, and O157. Our MLVA system had a relatively high resolution power for each of the three serogroups: Simpson's index of diversity was 0.991 (95% CI = 0.989-0.993), 0.988 (95% CI, 0.986-0.990), and 0.986 (95% CI, 0.979-0.993) for O26, O111, and O157, respectively. This system also detected outbreak-related isolates; the isolates collected during each of the 12 O26 and O111 outbreaks formed unique clusters, and most of the repeat copy numbers among the isolates collected during the same outbreak exhibited no or single-locus variations. These results were comparable to those of cluster analyses based on PFGE profiles. Therefore, our system can complement PFGE analysis-the current golden method. Because EHEC strains of three major serogroups can be rapidly analyzed on a single platform with our expanded MLVA system, this system could be widely used in molecular epidemiological studies of EHEC infections.

[Apoptosis effects on human esophageal cancer cells by soyasaponin Bb and its machanism].

OBJECTIVE: To explore the effect and potential mechanism of apoptosis in the Eca-9706 cells induced by soyasaponin Bb. METHODS: Different concentrations of soyasaponin Bb were on the Eca-9706 cell. Cell growth suppression rate was detected by MTT. Apoptotic rate was detected by TUNEL. The expression changes of HDAC1, NF-kappaB, PTEN, cyclin D1, c-met, VEGF or caspase-3 in the SSBb were detected by immunohistochemistry and immunoblotting. RESULTS: Compared with the control group, the growth-inhibition rate and apoptotic rate were increased, the expressions of PTEN, caspase 3 were increased, the expressions of c-met, VEGF, HDAC1, NF-kappaB and cyclin D1 were decreased. CONCLUSION: SSBb can suppress Eca-9706 cell growth. SSBb can exhibit reverse effects on over expression of c-met, VEGF in Eca-9706 cells. Eca-9706 cell apoptosis can be induced by SSBb through inhibiting HDAC1-NF-kappaB and activating PETEN and caspase-3 signaling pathways.

[Epidemiologic and clinical characteristics of 59 persons suspecting of being infected by the HIV virus despite having repeated negative laboratory findings].

OBJECTIVE: To describe the epidemiologic characteristics and clinical manifestations of 59 persons recruited via an internet chat group who complained of AIDS-like symptoms, so as to formulate effective intervention strategies and measures. METHODS: Case was defined as onset of any three of the following self-reported AIDS-like symptoms in a member of relevant "internet chat groups": persistent low grade fever, rash, swollen lymph node, fatigue, diarrhea, weight loss and low CD4(+)T count. We administered an internet-based questionnaire, and invited 59 of the 88 case-persons for voluntary physical examination and laboratory testing. RESULTS: The 59 case-persons came from 22 provinces; 54 (91.5%) were men; the median age was 34 (range: 22-53) years; 84.7% of them had high-risk sexual behaviors before the onset of self-reported symptoms. The median time interval from exposure to onset was 15 d (range: 1-365 d). Blood specimens for all the 59 case-persons were tested negative for HIV and syphilis antibodies. There was also no evidence of Xenotropic murine leukemia virus-related virus infection. One case-person was tested positive for hepatitis C virus antibody. The average CD4(+)T lymphocyte count was 707/µl. Of the 59 case-persons, 57 (96.6%) sought medical care from multiple providers; 40 were diagnosed to have no physical disorders. CONCLUSION: None of the 59 case-persons had any evidence of infection with HIV or any other infectious agents that could explain their self-reported symptoms.

[Effect of proliferation, c-met and vascular endothelial growth factor expression in the Eca109/9706 cells by liposomal quercetin].

OBJECTIVE: To explore the effect of cell proliferation, c-met and vascular endothelial growth factor (VEGF) expression by liposomal quercetin in the Eca109/9706 cells induced by liposomal quercetin. METHODS: The suppressing rate of cultured Eca109/9706 cells was detected by MTT assay. After 48-hour-exposure to liposomal quercetin, the expression and cellular localization of c-met and VEGF in Eca109/9706 cells were examined by using immunohistochemistry assay and Western blotting. RESULTS: The suppressing rate of Eca109/9706 cells was in the order of LQ2 group > LQ1 group > nLQ group > control group (P < 0.05). The protein expression of c-met and VEGF was found in Eca109/9706 cells cytoplasma as well as in nuclei by using immunohistochemistry assay. The Immuno-reaction intensity of c-met and VEGF was in the order of control group > nLQ group > LQ1 group > LQ2 group (P <0.05). The western blotting showed the intensity of immunoblotting signals for c-met-IR and VEGF-IR was in the order of control group > nLQ group > LQ1 group > LQ2 group (P < 0.05). CONCLUSION: The liposomal quercetin could suppress the proliferation of culture Eca109/9706 cells, which was about to be related with the suppression high expression of c-met and VEGF.

p21 is required for atRA-mediated growth inhibition of MEPM cells, which involves RAR.

All-trans retinoic acid (atRA), a metabolite of vitamin A, is essential for embryonic development. Thus the spatial and temporal dispersal of RA must be tightly controlled. Previous studies show that excessive atRA led to growth inhibition and p21 accumulation in mouse embryonic palatal mesenchymal (MEPM) cells. We reported here the identification of p21 as a required mediator during atRA-induced growth inhibition. atRA caused a G1 arrest in the cell cycle with an increase in the proportion of cells in G0/G1 and a decrease in the proportion of cells in S phase. In addition to a marked effect on cell cycling, atRA also triggered DNA fragmentation, reflected by an increase of the fraction of cells in the sub-G(1) population. Western blot analysis revealed that atRA treatment led to an increase in p21 level and a decrease in cyclin D1 protein and Rb phosphorylation. Using luciferase assay with reporter gene regulated by p21 promoter, we showed that atRA increased the reporter activity in a dose-dependent manner; and p21 siRNA blocked the growth inhibition by atRA, suggesting that p21 is required for atRA-mediated growth inhibition. Moreover, the induction of p21 by atRA was partially attenuated when RAR was silenced with specific siRNA. atRA stimulated RARE-driven reporter gene activity dose-dependently. Using chromatin immunoprecipitation, we demonstrated that RAR protein could bind to the p21 promoter. Taken together, our results indicate p21 is responsible for atRA-induced growth inhibition of MEPM cells and RAR plays a role during this process.

Molecular characterization of enterohemorrhagic Escherichia coli O157:H7 isolates dispersed across Japan by pulsed-field gel electrophoresis and multiple-locus variable-number tandem repeat analysis.

We identified seven distinct subtypes of enterohemorrhagic Escherichia coli (EHEC) O157:H7 isolates that were derived from sporadic cases and outbreaks from multiple prefectures in Japan in 2005. A surveillance system utilizing pulsed-field gel electrophoresis (PFGE), PulseNet Japan, was used. Some strains showed indistinguishable PFGE patterns using another restriction enzyme (BlnI or SpeI) in each subtype of EHEC O157:H7 isolates that were routinely subtyped by the XbaI PFGE pattern. In order to examine the genotypic relatedness of these strains, we carried out a multiple-locus variable-number tandem repeat (VNTR) analysis (MLVA). By using the MLVA system, we found that three of seven subtypes of EHEC O157:H7 strains that were isolated from sporadic cases dispersed across multiple prefectures within a few months showed indistinguishable PFGE patterns and identical MLVA types. Strains belonging to the other four subtypes of EHEC O157:H7 in the PFGE analysis were further classified into different clusters of EHEC O157:H7. Therefore, compared to PFGE, MLVA showed greater discriminatory power with respect to analysis of the isolates in this study.

[The mechanisms and effects of lutein on inducing the cell differentiation of human esophagus cancer EC9706].

OBJECTIVE: The purpose of the study was to explore the effects and molecular mechanisms of lutein on the differentiation of esophagus cancer EC9706 cell. METHODS: EC9706 cells were seeded in 1640 medium before the addition of test compounds. The respective test compound was added in fresh medium and the control cell received the vehicle (DMSO) or Fluorouracil. The proliferation and cell cycle of EC9706 were determined by MTT assay and flow cytometry, respectively. The change in cytomorphology was investigated by using HE staining. Proliferation and differentiation cells were checked and observed by methyl green-pyronine staining. The protein expression of cyclin D1 was detected by immunohistochemistry. RESULTS: Compared with the DMSO control group, the proliferation of the EC9706 cells treated with lutein (100 microg/mL and 150 microg/mL) could markedly be decreased and the cell cycle was blocked at G0/G1 phage which caused significant changes in the cytomorphology of EC9706 cell line, and the cell malignant degree tended to drop down, the protein expression of cyclin D1 was also down-regulated significantly. CONCLUSION: Lutein can inhibit the proliferation of EC9706 cell, and promote the cancer cell differentiation. cyclin D1 may be involved in cell proliferation and differentiation events in esophageal cancer EC9706 cell, which is regulated by lutein.