Background. Though Helicobacter pylori (HP) infections have progressively declined throughout most of the industrialized countries, a gradual increase in failure of HP eradication treatments is observed. Aim. To critically review evidence on the efficacy of the therapeutic availabilities for HP eradication, as yet. Methods. A selection of Clinical Trials, Systematic Reviews and Meta-analyses within the time period 2010–2012, was performed through a Medline search. Previous references were included when basically supporting the first selection. Results. An increasing rise in HP resistance rates for antimicrobial agents is currently observed. Further causes of HP treatment failure include polymorphisms of the CYP 2C19, an increased body mass index (BMI), smoking, poor compliance and re-infections. Alternative recent approaches to standard triple therapy have been attempted to increase the eradication rate, including bismuth-containing quadruple therapy, non-bismuth containing quadruple therapy, sequential therapy and levofloxacin-containing regimens. Conclusions. The main current aims should be the maintenance of a high eradication rate (>85%) of HP and the prevention of any increase in antimicrobial resistance. In the next future, the perspective of a tailored therapy could optimize eradication regimens within the different countries. Helicobacter pylori (HP) is an ubiquitous gram-negative bacterium. Its specific microbiological features allow its survival in the stomach and the colonisation of the antrum. HP expresses high levels of urease enzyme, producing NH3 and CO2 from the urea being present in the gastric juice [Helicobacter pylori infects approximately 50% of the adult population [ic juice . This meic juice . There aic juice . Helicobpulation , 6: onlyThis paper is conceived to critically review evidence on the efficacy of the therapeutic availabilities for HP eradication, available as yet. A Medline search was performed indicating as keywords “Helicobacter,” “Eradication Regimens,” “Eradication Rate,” “Epidemiology,” “Drug Resistance.” A selection of relevant English-language clinical trials, systematic reviews, and meta-analyses within the time period 2010–2012 was performed. Previous references were included when basically supporting the first selection. PubMed, Cochrane Library, Trip Database, and Ovid Medicine were used as referral databases.Jadad Scale (score 0 to 5) was used as a predetermined strategy of inclusion for clinical trials.Over recent decades, HP infections have progressively declined throughout most of the industrialized countries. HP still represents, by the way, a clinical issue among those people who were infected as children at a time when HP was endemic or among immigrants coming from endemic countries , 15. ManAt the same time, a gradual increase in failure of HP eradication treatments is found  and the Since resistance rates for antimicrobial agents rise with their indiscriminate use, clarithromycin (CAM) resistance may be due to the widespread use of this agent for upper respiratory tract infections , 20. De P < 0.00001) and a lower overall cost. Antimicrobial susceptibility testing is therefore recommended before first-line treatment for HP infection. Another important factor is polymorphism of the CYP2C19 isoenzyme [The prevalence of antimicrobial resistance might be significantly different between countries, which will greatly affect eradication rates , 31. Forsoenzyme , 36 thatsoenzyme . Gender soenzyme . The maiThe possibility of a tailored therapy will be a chance in the next future to optimize the most effective eradication regimen within the various geographical areas; for this proposal, the determination of HP genotype from gastric tissue samples or stools as well as the determination of CYP2C19 polymorphism is required .The most widely recommended treatment in international guidelines for the eradication of HP is the Standard Triple Therapy based on a proton-pump-inhibitor (PPI) to which two antibiotics (CAM plus amoxicillin (AMPC or MNZ) are associated for 7 days –41. WithAn efficacy of ranitidine bismuth-based triple therapies has also been demonstrated as retreatments . Non-bisH. pylori resistant to both CAM and MNZ, with an ITT of 76% [Because of the scarce compliance to these regimens as well as the increased resistance to both CAM and MNZ, the “sequential therapy” was introduced as a promising new treatment comprising 5 days of amoxicillin with a PPI, followed by 5 days of the same PPI but with a nitroimidazole  together with CAM. The sequential therapy was largely experimented in Italy where an increase in the eradication rate by at least 10% was observed, compared with standard regimens, independently on clarithromycin resistance , 57. ThiT of 76% . A studyT of 76% , thus suT of 76% . What fiT of 76% . The nonT of 76% .H. pylori infection is a PPI, AMPC, and CAM for 7 days [H. pylori CagA status for the efficacy of antibiotic treatment considering an infection with cytotoxic strains as a good predictive marker of successful therapy [S. boulardii and L. reuteri as an adjunct to HP eradication treatment has recently been established [In a recent trial by Basu et al.  a new rer 7 days . Accordir 7 days . Some au therapy –13, 66. ablished , 68. standard triple therapy with PPI (1-2 mg/kg/day), AMPC (50 mg/kg/day), CAM (20 mg/kg/day), or MNZ (20 mg/kg/day); bismuth-based quadruple therapy: bismuth subsalicylate (8 mg/kg/day), AMPC (50 mg/kg/day), and MNZ (20 mg/kg/day); sequential therapy: PPI (1-2 mg/kg/day) AMPC (50 mg/kg/day) for 5 days, then PPI (1-2 mg/kg/day), CAM (20 mg/kg/day), and MNZ (20 mg/kg/day) for 5 days. In 2011 the European and North American Societies of Pediatric Gastroenterology Hepatology and Nutrition (ESPGHAN and NASPGHAN) emitted evidence-based guidelines for managing HP in children . These gAs for adults, an antibiotic susceptibility test should be performed whenever a high resistance rate (>20%) in the area is known. Once treatment is completed a non invasive test should be done to ensure eradication .Over the past two decades, HP has represented a pivotal role in nonmalignant diseases of the upper gastrointestinal tract, as peptic ulcer disease (PUD), functional dyspepsia (FD), and gastroesophageal reflux disease (GERD) . IrrespeAs regards FD (as defined by Rome III criteria) a possible role of HP still remains uncertain . The patVirulence determinants of HP may be connected to gastric carcinogenesis; the use of genomewide germline analyses might offer the chance to identify high-risk groups of gastric cancer . Many different factors play a role in the development of HP infection, including geography, ethnicity, age, and socioeconomic elements. The efficacy of various available treatment strategies varies significantly within different regions. A progressive decline in infection rates has been secondary to the improvements in sanitation as well as to the use of combination anti-HP regimens over the past decades. The gradual increase in failure of HP eradication treatments is mainly secondary to antimicrobial resistance, in particular to clarithromycin.Quadruple therapy (PPI + CAM + AMPC and/or MNZ ± Bismuth) and sequential therapy  are efficacious alternatives to standard triple therapy for both adults and children. The main current effort should be to maintain the eradication rate of HP above 85% and to prevent any increase in antimicrobial resistance in the general population. The possibility of a tailored therapy will be a chance in the next future to optimize the most effective eradication regimen within the various geographical areas.


This study is aimed to evaluate the clinical significance of heart rate turbulence (HRT) parameters in predicting the prognosis in patients with chronic heart failure (CHF).From June 2011 to December 2012, a total of 104 CHF patients and 30 healthy controls were enrolled in this study. We obtained a 24-hour Holter ECG recording to assess the HRT parameters, included turbulence onset (TO), turbulence slope (TS), standard deviation of N-N intervals (SDNN), and resting heart rate (RHR). The relationships between HRT parameters and the prognosis of CHF patients were determined.1/2 rate than those with NYHA grade II/III. There were statistical differences in TS, LVEF, SDNN and RHR between the non-deteriorating group and the non-survivor group. Significant differences in TS among the three groups were also found. Furthermore, CHF patients in the non-survivor group had lower levels of TS than those in the deteriorating group. Correlation analyses indicated that TO negatively correlate with SDNN, while TS positively correlated with SDNN and left ventricular ejection fraction (LVEF). We also observed negative correlations between TS and left ventricular end-diastolic cavity dimension (LVEDD), RHR, homocysteine (Hcy) and C-reactive protein (CRP). Multivariate Cox regression analysis further confirmed that LVEF (≤30%), HRT2, SDNN and RHR were independent risk factors which can indicate poor prognosis in CHF patients.The assessment follow-up period lasted until January 31, 2013. The overall mortality of CHF patients was 9.6% (10/104). Our results revealed that CHF patients had higher levels of TO than those of healthy subjects, but the TS levels of CHF patients were lower than that of the control group. CHF patients with NYHA grade IV had higher HRTOur findings indicate that HRT may have good clinical predictive value in patients with CHF. Thus, quantifying HRT parameters could be a useful tool for predicting mortality in CHF patients. Chronic heart failure (CHF), one of the most common forms of heart failure, is a pathophysiological state in which an abnormality of cardiac function is responsible for the failure of the heart to pump blood at a rate necessary to deliver nutrients to metabolizing tissues . Despitein vitro stimulation, and cannot reflect the instantaneous malignant arrhythmias. [Clinically, the currently accepted prognostic indicators for CHF mainly consist of heart rate variability (HRV) and LVEF, which may both have some limitations to their own extents . HRV, asythmias. . As for ythmias. ,12. TherHeart rate turbulence (HRT), modulated by the baroreceptor reflex, is a physiological biphasic response of the sinus node to premature ventricular contractions, which involves a short initial acceleration followed by a deceleration of the sinus rhythm due to the baroreflex.  There arThe Ethics Committee of the First Affiliated Hospital of Harbin Medical University approved the study design. All patients gave informed consent in written to undergo diagnostic and therapeutic procedure at the time of hospitalization.The study was performed in patients diagnosed of CHF when admitted to the First Affiliated Hospital of Harbin Medical University from June 2011 to December 2012. Patients enrolled in this study must satisfy all of the following criteria: (1) all patients in the study had to meet the diagnostic criteria for CHF as specified by the 1995 World Health Organization/International Society and Federation Cardiology (WHO/ISFC); [In our present study, 104 CHF patients  were enrolled with a mean age of 61.0 ± 10.0 years . According to the NYHA classification, these patients were divided into grade II , grade III  and grade IV  groups. There were 37 patients  with ischemic heart disease and 67 patients  with nonischemic heart disease. In addition, 30 healthy controls  with a mean age of 58.0 ± 9.0 years  were enrolled in the control group after detailed medical examination, which includes medical history taking, physical examination, chest X-ray, echocardiography, Holter monitoring and blood biochemical examination.0 group with TO < 0 and TS > 2.5 ms/normal-to-normal interval ; HRT1 group with either TO ≥ 0 or TS = 2.5 ms/RR ; HRT2 group with TO ≥ 0 and TS ≤ 2.5 ms/RR  [All patients were managed under a uniform and standard assay method. HRT of CHF patients were quantified by TO and TS acquired through the 24-hour 3-lead Holter digital recordings  according to a previously published method . HRT wasbnormal) . For thebnormal) .Two-dimensional and Doppler echocardiographic examinations were performed with a HP5500 ultrasound system . In the parasternal long-axis view, M-mode echocardiograms were made of left ventricle from the left hemithorax in short-axis.The median duration of the entire study follow-up was 10.5 ± 8.3 months . All events recorded in the study were adjudicated blindly by an ad-hoc committee on the basis of pre-agreed definitions and procedures. Follow-up information was collected for all patients through outpatient service and phone calls. The end point was prospectively defined as the composite of heart failure and cardiac mortality.The assessment follow-up period lasted until January 31, 2013. Finally, the overall mortality of CHF patients was 9.6% (10/104). There were 32 CHF patients  exhibiting a deterioration of cardiac function and 62 patients showing no significant deterioration.2 or Fisher’s exact test for dichotomous parameters. Correlation between autonomic markers was assessed with Spearman correlation coefficient. Survival curves were estimated with the Kaplan-Meier method and compared by the log-rank test. Independent predictors of death were identified with multivariate Cox proportional hazards model and expressed as hazard ratio (HR) with 95% confidence interval (CI). A P value of less than 0.10 was considered statistically significant and all tests were two-sided. All analyses were performed using the SPSS 18.0 software .Data were expressed as mean ± standard deviation (mean ± SD) unless otherwise specified and qualitative data are presented as percentages. Comparative analyses between the groups were performed with the Student’s t-test for continuous variables and ΧP < 0.001), but the TS levels of CHF patients were lower than that of the control group . There was a significant difference in the rate of HRT1/2 between CHF patients and healthy controls . Furthermore, CHF patients with NYHA grade IV had higher HRT1/2 rate than those with NYHA grade II/III  , but no associations were found between TO and other factors . TS positively correlated with SDNN  and LVEF . However, there were negative associations between TS and LVEDD , RHR , Hcy  and CRP . We also observed no correlations with TS to age and TC levels (both P > 0.05). Correlations of TO and TS with relevance factors are shown in Table Correlation analyses indicated that TO negatively correlated with SDNN . Furthermore, CHF patients in the non-survivor group had lower levels of TS than those in the deteriorating group (P < 0.001).Our results suggested that there were statistical differences in TS, LVEF, SDNN and RHR between the non-deteriorating group and the non-survivor group    , LVEF ≤30%), SDNN (≤100 ms), RHR (≥75 bpm), HRT5) Table . Multiva, LVEF , HRT2, SDNN and RHR were independent risk factors for poor prognosis of CHF patients. Among these risk factors, HRT2  was the strongest predictor of CHF with a high hazard risk of 5.12 after CHF and remained highly significant after adjustment for LVEF and other clinical risk factors. The combination of abnormal TO and TS may result in tachycardia which would cause decreased heart efficiency in increased myocardial oxygen consumption, decreased myocardial perfusion during myocardial relaxation and diastolic, thereby leading to myocardial injury and aggravating or worsening heart failure [Generally, HRT is described as the acceleration and subsequent deceleration of sinus rate . Abnorma studies . In addi studies . Consist studies . Univari failure . NeverthOur findings indicated that HRT may have good clinical predictive values in patients with CHF. Thus, quantifying HRT parameters could be useful tool in predicting mortality in CHF patients. However, due to the limitations mentioned above, further detailed studies with larger sample size and longer follow-up periods are still required to confirm our results.The authors have declared no competing interests among themselves.YDC carried out the experiments and drafted the manuscript. WSJ, GS and FW analyzed the data. XHY, SL and QW contributed the material and the analysis tools. YDC and QXF participated in the design of the study and performed the statistical analysis. QXF conceived the study, and participated in its design and coordination and helped draft the manuscript. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2261/14/50/prepub


Cytotoxicity of laboratory-made functionalized multiwalled carbon nanotubes (CNTs) , compared to pristine MWCNTs, carbon black, and silica, has been assessed in human A549 pneumocytes by MTT assay and calcein/propidium iodide (PI) staining. Purity and physicochemical properties of the test nanomaterials were also determined. Subsequently, pulmonary toxic effects were assessed in rats, 16 days after MWCNTs i.t. administration (1 mg/kg b.w.), investigating lung histopathology and monitoring several markers of lung toxicity, inflammation, and fibrosis. In vitro data: calcein/PI test indicated no cell viability loss after all CNTs treatment; MTT assay showed false positive cytotoxic response, occurring not dose dependently at exceedingly low CNT concentrations (1 μg/mL). In vivo results demonstrated a general pulmonary toxicity coupled with inflammatory response, without overt signs of fibrosis and granuloma formation, irrespective of nanotube functionalization. This multitiered approach contributed to clarifying the CNT toxicity mechanisms improving the overall understanding of the possible adverse outcomes resulting from CNT exposure.Increasing application of engineered nanomaterials within occupational, environmental, and consumer settings has raised the levels of public concern regarding possible adverse effects on human health. We applied a tiered testing strategy including (i) a first  Nanotechnology is one of the fastest emerging fields involving development and manipulation of materials ≤100 nm in size. There are numerous potential perspectives for the applications arising from nanotechnology which include their use in a wide range of fields, for example, medicine, environment, occupational setting, chemistry, energy production, information and communication, heavy industry, and consumer goods. Human exposure to engineered nanomaterials (ENMs) can occur at different stages of their life cycle  and several concerns have been expressed about their potential to cause unanticipated adverse effects in humans.The remarkable diversity of engineered nanomaterials (ENMs), together with their unique properties and behaviour, complicates their risk assessment; there are currently about 50,000 different types of carbon nanotubes obtained by different raw materials and production processes. Similar diversity applies to other types of ENMs, rendering ad hoc risk assessment of all of these materials an immense task , 2.The REACH Regulation  is the current regulatory framework for chemical risk assessment and management in European Union. Although REACH should be applied to ENMs, the Technical Guidance Documents  for prepNevertheless, though particular ENMs properties and behaviour  have been described in several studies , severaFurthermore, there are major challenges in assessing exposure . ENMs are produced from many substances, in many forms and sizes, and with a variety of surface coatings. The health risk assessment for such diverse materials requires validated analytical methods permitting their characterization in bulk samples and their detection and measurement in workplace air . EffortsToday, there are not enough studies to validate the potential hazards posed by these novel materials and hence the definitive conclusions and tools, technologies, systems, and methods to obviate the risks. Several tools for the assessment of risks are still in the conceptual stage, and further, there are considerable uncertainties on how to assess nanoparticles (NPs) exposure  and what methods to use to assess toxicity.Thus, being unable to perform a quantitative risk assessment for ENMs, due to the lack of sufficient data on exposure, biokinetics, and organ toxicity, it should be made mandatory to prevent exposure by appropriate precautionary measures and practicing best industrial hygiene to avoid future shock scenarios from environmental or occupational exposures .In the safety assessment area, two basic questions still need to be addressed: (i) do nanomaterial properties necessitate a new toxicological science? and (ii) what are the biological and biokinetic properties to be considered for toxicity testing; in particular, can biology and certain mechanisms of effects of ENMs  be used as a basis for studying NPs toxicity and risk assessment? Addressing these questions is of crucial importance to define adequate strategies and establish whether ENM-tailored testing methods should be added to conventional toxicity testing protocols to comply with regulatory demand and properly characterize the ENMs potential hazard. physicochemical characterization , assisted by using reference materials, as well as the use of acellular systems to explore the reactivity of the materials in acellular environments. Subsequent steps include the use of validated cellular (in vitro) models to support evidence-based testing process, followed by a limited series of in vivo studies guided by information generated from in vitro studies, especially the toxicological data relevant to ranking of ENMs, designing appropriate exposure concentrations and defining the critical health endpoints to be monitored.According to the major institutions , 9, 10 aBiosafety should be evaluated by tests examining general toxicity, target organ toxicity, and biocompatibility in line with regulatory requirements, limiting the use of lab animals in toxicological research , 13, to in vitro and in vivo experiments. Such investigation is related to our recent studies on the pulmonary toxicity of a series of functionalized multiwalled carbon nanotubes (MWCNTs).The present work represents an example of a tiered approach for toxicity testing, employing CNTs as a practical model suitable to validate a two-level strategy combining in vitro the cytotoxicity of MWCNTs, with different degrees of functionalization, using MTT assay and calcein/propidium iodide (PI) staining. Human A549 pneumocytes, selected as potential target cells during respiratory exposure associated with occupational setting and environment , 14–18, β1), (2) interleukin-6 (IL-6), and (3) collagen (Type I) investigated by immunochemical methods, as markers of lung toxicity, inflammation, and fibrosis, respectively.Subsequently, the pulmonary toxic effects were assessed in rats 16 days after i.t. administration of the previously reported type of CNTs (1 mg/kg b.w.). Major endpoints tested included (i) histopathology of lung tissue (haematoxylin/eosin staining), (ii) apoptotic/proliferating features examined by TUNEL and PCNA immunostaining, and (iii) presence/distribution of (1) transforming growth factor-beta1   were obtained from Cheap Tubes Inc.  and CB from Aldrich-Fluka . SiOGermany) .2). MWCNTs with different functionalization were prepared as described by Fagnoni et al., 2009 . The labin vitro and in vivo exposure, exhaustive physicochemical characterization of differently functionalized CNTs was performed by IR spectroscopy, thermogravimetric analysis (TGA), and by noncontact atomic force microscopy (NC-AFM).Before the 2.It has been estimated that one functional chain is anchored each ~16 carbon atoms of the CNTs surface for the MW-COOH and each ~100 carbon atoms of the CNTs surface for the MW-NH2) and 500–2000 nm for the pristine MWCNTs. Other physicochemical characteristics are detailed in All the three types of CNTs had an outer diameter of 20–30 nm and the wall thickness was 1-2 nm. The particle length was 100–300 nm for both functionalized CNTs  analysis. Results indicated the presence of Fe and Ni impurities at concentrations of 0.25% and 1.5%, respectively, in pristine MWCNTs. Considerably lower metal concentrations were found in the functionalized nanotubes. In these materials, the content of Fe and Ni was 0.05%–0.15% and 0.3%–0.5%, respectively.The content of iron (Fe), cobalt (Co), and nickel (Ni) in both 2, SiO2, and CB) were prepared in DMEM plain or in NaCl 0.9% for the in vitro expo pure and in vivo exposure, respectively. Just before use, these suspensions were sonicated for 15 min with an ultrasonic Sonopuls  in a short break every two minutes, also vortexing the suspension on ice to further force the CNT dispersion, avoiding the agglomeration and the formation of bundle-like structures. No surfactants or solvents were used. The suspensions of the test materials were immediately used for the treatment. Previous to the experiments, a stock of 1 mg/mL and a suspension of 1 mg/kg b.w. of each test material . A549 cells from a human Caucasian lung adenocarcinoma with the alveolar type II phenotype were obtained from ECACC . The cells were cultured in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal calf serum (FCS), 2 mM L-glutamine, 50 IU/mL penicillin, and 50 At the end of the incubation period, cell cultures were examined by phase contrast microscopy using a Zeiss Axiovert 25 light microscope.4/cm2 in complete medium. After 24 h of cell attachment, plates were washed with 100 μL/well of phosphate-buffered saline (PBS). Cells were then exposed to suspensions (10 μL) of the test materials  at concentrations as 1, 10, and 100 μg/mL, for 24 or 48 h. No fetal bovine serum was used in these preparations as it was proven to interact with nanotubes [Cells were seeded in 96-well plates at a density of 1  × 10anotubes .μg/mL) and the time points of determinations (24 and 48 hr) have been derived from previous studies showing different adverse effects  caused by various carbon nanotubes.The doses tested (1–100 P < 0.05) were determined by one-way analyses of variance (ANOVA) (software package SPSS Inc. 1999).Six replicate wells were used on each 96-well plate for all the treatments, and each experiment was repeated independently three times. Identical treated cultures were taken as replicate measure for statistical tests. Significant effects -Dead (propidium iodide-red fluorescence) staining  and the MTT assay were used to assess cytotoxicity. The live/dead viability test detects cell membrane integrity and measures the number of damaged cells. Fluorescence microscopy labels both live (green colour) and dead (red colour) cells simultaneously thereby permitting visualization and enumeration of all cells by a single counting procedure. Viability was expressed as a percent of the total number of cells counted.The MTT assay uses tetrazolium salts to assess mitochondrial dehydrogenase activity. Only active mitochondria contain dehydrogenases enzymes able to cleave the tetrazolium ring and reduce MTT to insoluble dark-blue formazan crystals and, therefore, the reaction only occurs in viable cells. Absorbance, directly proportional to cell viability, was determined at 550 nm in a Biorad microplate reader. The absorbance values were normalized by the controls and expressed as percent viability.After the incubation period, all cultures were also examined and photographed in phase contrast using a Zeiss Axiovert 25 light microscope combined with a digital camera (Canon Powershot G8).All experimental procedures involving animals were performed in compliance with the European Council Directive 86/609/EEC on the care and use of laboratory animals.Adult Sprague-Dawley rats  were purchased from Charles River Italia  at least 2 weeks before treatment and allowed to acclimatize for 3 weeks. Throughout the experiment, animals were kept in an artificial 12 h light : 12 h dark cycle with humidity at 50 ± 10%. Animals were provided with rat chow (4RF21 diet) and tap water ad libitum.For the treatment, groups of 6 rats were anesthetized with pentobarbital sodium for veterinary use and were intratracheally instilled with MWCNTs, pristine or functionalized, dispersed at a dose of 1 mg/kg b.w. in NaCl 0.9% see . μL/100 g b.w.); lung preparation for microscopic evaluations was done by vascular perfusion of fixative. Briefly, the trachea was cannulated, and laparotomy was performed. The pulmonary artery was cannulated via the ventricle, and an outflow cannula was inserted into the left atrium. In quick succession, the tracheal cannula was connected to about 7 cm H2O pressure source to inflate the lungs with air, and clearing solution  was perfused via the pulmonary artery. After blood was cleared from the lungs, the perfusate was switched to fixative consisting of 4% paraformaldehyde in 0.1 M phosphate buffer (pH 7.4). After fixation, the lungs were carefully removed, washed in NaCl 0.9%, and postfixed by immersion for 7 h in 4% paraformaldehyde in 0.1 M phosphate buffer (pH 7.4); then, the tissues were dehydrated through a graded series of ethanol and finally embedded in Paraplast. Eight μM thick sections of the samples were cut in the transversal plane and collected on silane-coated slides.Sixteen days after the i.t., treated and control rats were deeply anesthetized with an overdose i.p. injection of 35% chloral hydrate  for histological examination.β1), (ii) interleukin-6 (IL-6), (iii) collagen (type I), and (iv) proliferating cell nuclear antigen (PCNA-PC10), as typical markers of general lung toxicity, inflammation, fibrosis, and cell proliferation, respectively.Immunohistochemistry was performed using commercial antibodies on rat lung specimens, to assess the presence and distribution of (i) transforming growth factor-beta1  diluted 1 : 100, (ii) a primary rabbit polyclonal antibody against collagen (type I)  diluted 1 : 400, (iii) a primary goat polyclonal antibody against IL-6 (Santa Cruz Biotechnology) diluted 1 : 100, (iv) a primary mouse monoclonal antibody against PCNA  diluted 1 : 5 in PBS. Biotinylated anti-rabbit, anti-goat, and anti-mouse secondary antibodies and an avidin biotinylated horseradish peroxidase complex  were used to reveal the sites of antigen/antibody interaction. 3,3′-Diaminobenzidine tetrahydrochloride  was used as the chromogen, followed by a light counterstaining with haematoxylin. Then, the sections were dehydrated in ethanol, cleared in xylene, and finally mounted in Eukitt .Lung sections were incubated overnight at room temperature with (i) a primary rabbit polyclonal antibody against TGFIn the case of negative controls, some sections were incubated with phosphate-buffered saline instead of the primary antibodies; no immunoreactivity was observed in these conditions.in situ detection of DNA fragmentation using the terminal deoxynucleotidyl-transferase (TUNEL) assay . The lung sections were incubated for 5 min with 20 μg mL−1 proteinase-K solution at room temperature, followed by treatment with 3% H2O2 to quench endogenous peroxidase activity. After incubation with the TUNEL solution (90 min with TdT/biotinylated dNTP and 30 min with HRP-conjugate streptavidin) in a humidified chamber at 37°C, the reaction was developed using 0.05% 3-amino-9-ethylcarbazole (AEC) in 0.1 M TRIS buffer (pH 7.6) with 0.2% H2O2; in some specimens the reaction was developed using a 0.1% DAB solution. In addition to morphological criteria, the apoptotic cell death was assayed by As a negative control, the TdT incubation was omitted; no staining was observed in these conditions.β1, and collagen (type I) was evaluated in conventional brightfield microscopy by recording the localization and intensity of labelling according to a semiquantitative scale from absent/undetectable (−) to maximal (++++). Then, to assess the significance of the immunohistochemical results, a Kruskal-Wallis nonparametric analysis of the semiquantitative data was performed. A P value of <0.05 was considered significant. (a) Scoring different specimens, the immunostaining for IL-6, TGFt-test, and differences between medians were considered significant at *P < 0.05.(b) The evaluation of PCNA- and TUNEL-cytochemically positive cells  was calculated as the percentage (Labelling Index) of a total number (about 500) of bronchiolar, alveolar, and macrophagic cells, for each animal and experimental condition, in different representative microscopic fields. Statistical analyses among the different biological situations were performed by the Student's The slides were observed and scored with a brightfield Zeiss Axioscop Plus Microscope. The images were recorded with an Olympus Camedia C-2000 Z digital camera and stored on a PC running Olympus software.2) were tested in comparison with pristine MWCNTs in cultures of A549 pneumocytes. Carbon black (CB) and silica (SiO2) were also investigated as reference nanoparticles [Laboratory-made functionalized multiwalled CNTs , with 50% viability loss at 10 μg/mL and no further cell death increase at 100 μg/mL  and CB the calcein/PI test indicated no loss of cell viability , whereas00 μg/mL . Similarμg/mL , core particle, and surface chemistry. The present data on (i) dose metric characteristics , while contrasting results were obtained from MTT and calcein/PI after pristine and functionalized MWCNTs. As far as physicochemical properties concern, although the functionalized nanomaterials possessed lower aggregate size and particle size ranges  ageristics , and  on some cell kinetic  and cytochemical parameters  investigated in rats after 16 days from a single i.t. exposure (1 mg/kg b.w.).Subsequently, the second focused pristine and lab-made MW-COOH and MW-NH2) already evidenced in the first in vitro step, as described previously and irrespectively of the nature of functionalization. After i.t. exposure to all the three different CNTs, lung morphology micrographs clearly showed a marked uptake of the CNTs into the macrophages . NoticeaHistologically, alteration of lung architecture was also observed in several areas showing collapsed thick-walled alveoli and presence of microhaemorrhagic foci , not accTUNEL and PCNA staining, employed as typical markers of apoptosis and cell proliferation, respectively, showed a significant increase of reactivity in different cell populations , as expression of (i) diverse sensitivity of different cell categories to the insult and (ii) an improved cellular turnover aimed at replacing damaged elements and finalizing a repair process Figures  and 6.β1, evidencing the cellular inflammatory reaction to the CNTs instillation  and oxidative stress , 45, 55,illation , while, The persistence of inflammation, still observable 16 days after instillation, could be explained as the consequence of failure to eliminate the inciting agent , as demonstrated by the durability of fibrocarbonaceous nanomaterial internalized by macrophages. Differently, the lack of a marked collagen accumulation could be attributable to a physiological tissue repair in which the tissue remodelling occurred moderately.These research studies on CNTs clearly indicated that certain negative properties that are typical of classic CNTs such as cytotoxicity, poor blood compatibility, inflammogenic effects, and target-organ toxicity are maintained to some extent in functionalized CNTs, supporting the view that overcoming the safety problems remains a considerable challenge for these nanomaterials.in vitro step investigation reported conflicting results on the cytotoxic effects of pristine and differently functionalized MWCNTs; specifically all the tested CNTs exhibited mild to moderate cytotoxicity when tested using MTT assay, with changes occurring not dose-dependently already at very low CNT concentrations. By contrast, the calcein/PI test showed that cell viability was unaffected by all tested CNTs, even at the higher concentrations and for longer exposure time. These diverging results highlighted some limitations intrinsical to classical in vitro cytotoxicity tests, applied to the study of the ENMs, due to the peculiar physicochemical characteristics of these new nanomaterials, fully supporting the notion [in vitro studies can be accepted in the safety evaluation of nanomaterials, calling for research approaches complementary to the in vitro studies aimed at understanding effects at all physiological levels and predicting human health hazards.In summary, our first e notion , 57 thatin vivo.Strategies for the safety assessment of ENMs and agglomerates are under discussion and development in several national and international projects. It has been suggested  to develin vivo study, being the subsequent second essential stage, has provided fundamental information, contributing to clarifying the CNT toxicity mechanisms and to understanding the different CNT toxicity targets, at different pulmonary cytochemical levels, in order to improve the overall understanding of the possible adverse outcomes resulting from CNT exposure.Since investigations in laboratory animals may give essential insight, the in vivo data demonstrated that intratracheal instilled MWCNTs can induce lung toxicity associated with inflammation, even in absence of fibrosis, irrespective of nanotubes functionalization. This latter finding suggests caution before considering chemical functionalization as a broad way to improve biocompatibility and safety characteristics of CNTs, being even more important for CNTs that are proposed for diagnostic or therapeutic applications.In particular, our These overall experimental results further support the use of a multitiered strategy for the safety assessment of the human health impact of novel nanotechnology-based products.


Cynomorium songaricum Rupr. (CS) is traditionally used to improve sexual function and treat kidney dysfunction in traditional Chinese medicine, although little is known about whether CS has effects on longevity. Here, we show that CS supplementation in the diet extends both the mean and maximum lifespan of adult female flies. The increase in lifespan with CS was correlated with higher resistance to oxidative stress and starvation and lower lipid hydroperoxides (LPO) levels. Additionally, the lifespan extension was accompanied by beneficial effects, such as improved mating readiness, increased fecundity, and suppression of age-related learning impairment in aged flies. These findings demonstrate the important antiaging effects of CS and indicate the potential applicability of dietary intervention with CS to enhance health and prevent multiple age-related diseases.Aging is highly correlated with the progressive loss of physiological function, including cognitive behavior and reproductive capacity, as well as an increased susceptibility to diseases; therefore, slowing age-related degeneration could greatly contribute to human health.  Age is the most important risk factor for the progressive loss of physiological function, increased susceptibility to diseases and mortality due to the gradual accumulation of damage at the cellular, molecular, and tissue levels . BecauseDrosophila and Caenorhabditis elegans [Curcuma longa (turmeric), has demonstrated protection against oxidative stress and aging in several animals [Accumulating evidence suggests that nutritional supplementation also increases lifespan and delays age-related syndromes. For example, catechin extracts, including epigallocatechin gallate (EGCG), from green tea prolong the lifespan and increase the antioxidative activity of  elegans , 7. Curc animals , 9. Resv animals . This sm animals . Approac animals , 13.Drosophila melanogaster, we identified a herbal medicine with a potential effect. Cynomorium songaricum Rupr. (CS), also known by its Chinese name, Suoyang, is a parasitic plant that grows in northwest China. Extracted from its stem is a widely used Chinese herbal remedy for treating sexual dysfunction, infertility, deficient kidney function, and lumbar weakness, as well as for facilitating catharsis, as described in ancient Chinese medical books. Chemical components identified in CS extracts include organic acids, triterpenes, flavonoids, polysaccharide, and steroidal compounds [Drosophila, which is an ideal organism for in vivo aging studies. Our data show significant extension of lifespan, improved resistance to environmental stress, and suppressed age-related syndromes, suggesting the possible clinical utility of CS in slowing the aging process.Given that natural compounds can decrease mortality, we considered an alternative strategy to identify traditional Chinese medicines that have prolongevity effects. In an unbiased screening test for a lower incidence of mortality in the ompounds –16. A reompounds . CS extrompounds , 19. Othompounds , 21. Yu ompounds . DespiteCanton S background and maintained in cornmeal standard media at 25°C under a 12 h light-dark cycle. Emerging adult flies were collected within 24 h and separated by sex. From early adulthood, flies were supplemented with CS as a concentrated herbal medicine purchased from Ko Da Pharmaceutical Co., Ltd.  under the good manufacturing practice (GMP) criteria. All experimental procedures were performed at 29°C with 50–60% relative humidity. CS powder was dissolved in H2O and added to cornmeal media at 10, 20, and 30 mg/mL concentrations, which were proportional to the effective daily dosage for human oral administration. In a lifespan analysis, twenty to thirty flies were raised in a food vial with or without CS; at least four vials were prepared for each treatment. Food vials were replaced every 2 to 3 days, and dead flies were counted at that time. At least three replicates were conducted per trial.The wild-type fly line used in this study was of the 2•−) that cause oxidative damage to animals, and we thus measured the resistance of the flies to this oxidative stress. Similarly, flies were transferred to vials containing a filter paper with hydrogen peroxide (H2O2)  in a 5% glucose solution to gauge the flies' resistance against H2O2-induced hydroxyl radical (OH•−) stress. We also kept flies in vials with wet papers only and evaluated their stamina against starvation-induced oxidative stress. Flies were scored for mortality at least three times daily until all were dead. Every 24 h, flies were transferred to new vials with freshly prepared solutions. At least three vials per treatment were performed, and all experiments were conducted in triplicate.Female flies were collected in a vial and pretreated with various doses of CS for 20 days at 29°C. Before testing, the flies were starved for 6 h. For oxidative stress assays, flies were transferred to food-free vials containing a filter paper saturated with 10 mM paraquat  diluted in a 5% glucose solution. Paraquat generates superoxide anions  activity using catalase and SOD assay kits . Briefly, female flies were reared on a control or experimental diet containing different doses of CS at 29°C for 20 days. For each enzyme activity assay, flies in each group were collected and homogenized in cold buffer. Supernatants were collected after centrifugation and assayed for catalase and CuZn-SOD enzyme activity according to the kit's protocol.LPO was evaluated by quantifying the malondialdehyde (MDA) level using a thiobarbaturic acid (TBA)  method . BrieflyVirgin females were collected and treated with different doses of CS for 10 or 20 days as experimental groups. The day of eclosion was designated as day 1 of adult life. To measure courtship behavior, one experimental female was paired with a virgin male, which was 9-day-old and had not received CS supplementation, in a plastic chamber. The experiment was conducted at Zeitgeber time (ZT) 3 to ZT6, which is the time that females show higher mating activities . ZT0 andn = 50–60).Virgin females fed varying doses of CS for either 10 or 20 days served as experimental groups. Ten experimental females were mated with an equal number of virgin males, which were 9-day-old and had not received CS, for 24 h in a vial. The mated flies were then transferred to a new vial for egg laying for 24 h, and the total eggs laid in the vials were counted manually. An index of fecundity was calculated as the mean daily egg production per female. At least five vials per treatment were performed (+) were paired with 60 V electric shocks and then received a second odor (CS‒) without shocks. Learning ability was determined immediately after training. To perform the test, the trained flies were trapped into the choice point of a T-maze in which they were exposed simultaneously to OCT and MCH. A performance index (PI) was calculated to represent the conditioned odor avoidance. A PI of zero indicated a 50 : 50 distribution, and a PI of 100 showed a 0 : 100 distribution away from the CS+ odor.Female flies were collected after eclosion and pretreated with 20 mg/mL CS for 20 days at 29°C. One training session was performed according to a previous study . BrieflyU-test. Other experimental comparisons were made by performing a Student's t-test compared with the control groups. Data are represented as the mean ± SEM, and a P value of <0.05 was considered significant.All experiments were conducted with at least three independent replicates, and statistical analyses were performed with SigmaStat v3.5 software . Data for lifespan extension and oxidative stress challenges represented experiments performed with a total number of flies ranging from 250–350, and the log-rank test was assessed for difference analysis. The copulation latency was not distributed normally, thus we used nonparamertric Mann-Whitney P < 0.001), and the maximum magnitude of increase in the mean lifespan was 1.18% at 20 mg/mL CS. Our results suggest that the addition of CS into the diet significantly extends lifespan. Because female flies were more responsive to CS, this sex was used for further experiments.To examine whether the application of CS can extend the fly lifespan, flies were supplemented with 10, 20, and 30 mg/mL CS immediately after adult eclosion at 29°C throughout the experimental procedure. Our data demonstrated that the lifespan extension caused by CS was gender-dependent. As shown in 2O2 which produce ROS to trigger cellular senescence and cell death. We found that CS protected females against paraquat toxicity and extended both the mean and maximum lifespan . In the H2O2 challenge, flies fed CS, especially at the 20 mg/mL dose, survived longer than untreated controls (P < 0.001), indicating that CS increased oxidative resistance against paraquat and H2O2 toxins. A similar trend showing increased resistance to starvation was also detected  but not in the 30 mg/mL CS group. These results reveal that treatment with CS can improve resistance to some oxidative stress, suggesting that the lifespan extension induced by CS might be associated with an increase in antioxidative defense.As shown in detected . The meaP < 0.05 and 0.01, resp.) and 61.8% for the 30 mg/mL CS diet (P < 0.01) compared with the control group. We also analyzed the levels of LPO, a process that refers to lipid oxidation caused by ROS, partly due to aging . CS supplementation not only increased antioxidant activity through a change in catalase activity but also decreased LPO formation, suggesting that CS protects tissue from ROS attacks.To characterize the mechanism by which CS protects against oxidative stress, we evaluated the changes in free radical scavenging enzymes in CS-feeding flies, such as catalase and SOD. The data revealed increased catalase but not CuZn-SOD activity in the CS-supplemented groups compared with the controls Figures . As Figuto aging . We obseP < 0.01) , and this result also appeared in females with a 30 mg/mL CS diet for 20 days   or 20 days (aged) was paired with a 9-day-old control virgin male in a plastic chamber to determine their copulation duration . The cop Figures . We also < 0.01) . Our obsP < 0.01, 0.05, and 0.05, resp.), but for young flies, the higher dose of CS (30 mg/mL) caused a moderate decrease in egg-laying ability   , suggestP = 0.0011), suggesting that CS has a beneficial effect on the maintenance of learning ability with age.While feeding CS to increase fly's longevity and oxidative resistance has been identified in our present study, the effects of these longevity-promoting factors on cognitive behavior are unknown. We attempted to ascertain whether CS administration could also improve learning behavior in aged flies. Because supplementation of CS at 20 mg/mL shows a significant effect on survival rate , flies e2O2, and starvation stress challenges. We also found that CS plays crucial roles in modulating mating readiness, reproductive capacity, and cognitive behavior. These results indicate that CS delays age-related syndromes during the aging process.This study shows for the first time that adult-onset CS supplementation in flies enhances longevity and increases resistance to paraquat, HWe initially investigated whether CS could prolong lifespan in both sexes of flies; unexpectedly, the lifespan extension induced by CS supplementation was only observed in females. The mean lifespan increased in females for all three tested doses, with the largest increase observed at 20 mg/mL CS , suggest2O2, which generate free radical superoxide anions and hydroxyl radicals, respectively. We also examined changes in antioxidative defense systems, especially catalase and CuZn-SOD. CS altered catalase enzyme activity but made no changes to CuZn-SOD, in contrast with what is observed for green tea extract, Rhodiola rosea, and Rosa damascene, which also extend fly lifespan [R. rosea and R. damascene failed to change these antioxidant enzymes. Our data suggest that CS may work via catalase pathway to protect flies against elevated oxidative stress with age.The underlying mechanisms by which CS extends the mean and maximum lifespan of flies remain obscure. Because aging is correlated with oxidative damage caused by ROS attacks, it is speculated that elevating oxidative resistance could yield a prolongevity effect , 32. We lifespan , 33, 34.Drosophila species D. pavani and D. gaucha, copulation duration varies with age, with older female flies showing longer copulation duration than younger flies [D. melanogaster, the copulation duration is also correlated with age. Beaver et al. found that 4-day-old males copulated for shorter durations than 2-day-old males [Because we found that the administration of CS in adults could decrease fly mortality and improve tolerance to oxidant stress, we examined whether CS could spawn other benefits, especially in courtship behavior and reproductive capacity, which are both negatively correlated with age . Copulater flies . In D. mld males . CompareR. rosea can live longer but show impaired fecundity [Prior studies have highlighted the negative correlation between lifespan extension and fecundity. Flies that undergo downregulation of the insulin signaling pathway, with DR conditioning, or supplemented with ecundity , 37, 38,ecundity , 39. Wooecundity . In contFunctional changes in synapses that contribute to altered neuronal plasticity and cognitive decline have a well-described association with age in mammals , 42. In Another mechanism for the retention of learning ability apparent in CS-fed aged flies may be correlated with enhanced antioxidant defense systems. A previous study indicated that CS has neuroprotective effects against staurosporine-induced apoptosis in human neuroblastoma cells . While sThe present results provide clues as to how CS could alleviate age-related syndromes, especially in terms of brain function, courtship behavior, and reproductive capacity, when CS is provided at a younger age. Our study provides some basic evidence supporting clinical treatment of replenishing the vital essence in traditional Chinese medicine and might provide valuable insight for developing new drugs or food additives to delay aging and the progression of age-related diseases.


Peptidyl-prolyl cis–trans isomerase NIMA-interacting 1 (PIN1) plays an important role in cancer development. The relationship between PIN1 −842G/C (rs2233678) polymorphism and cancer risk was inconclusive according to published literature.heterogeneity<0.01) and CG/CC  genotypes were associated with a significantly reduced cancer risk compared with those with wild GG genotype. Sub-group analysis revealed that the variant CG  and CG/CC  genotypes still showed an reduced risk of cancer in Asians; while no significant association was observed in Caucasians . Furthermore, sensitivity analysis confirmed the stability of results. Begg's funnel plot and Egger's test did not reveal any publication bias.A literature search, up to February 2013, was carried out using PubMed, EMBASE and the China National Knowledge Infrastructure (CNKI) database. A total of 10 case-control studies including 4619 cases and 4661 controls contributed to the quantitative analysis. Odds ratio (OR) and 95% confidence intervals (95% CI) were used to assess the strength of association. Overall, individuals with the variant CG (OR = 0.728, 95% CI: 0.585,0.906; PThis meta-analysis suggests that the PIN1 −842G/C polymorphism is associated with a significantly reduced risk of cancer, especially in Asian populations. Pro-directed phosphorylation is a critical signaling mechanism in various cellular processes, including transcription, RNA processing, cell cycle progression, cell proliferation and differentiation The gene that encodes PIN1 protein is mapped to chromosome 19p13.2. Several studies have investigated the relationship between the single nucleotide polymorphisms  in the PIN1 promoter region and risk of cancers, such as breast cancer To confirm the association between −842(G>C) polymorphisms of PIN1 gene and cancer risk, we performed this meta-analysis by pooling all eligible studies to calculate the estimate of overall cancer risk and evaluate influence of cancer types and ethnicity.A literature search was carried out using PubMed, EMBASE and China National Knowledge Infrastructure (CNKI) database up to February 2013. There was no restriction of origin or languages. Search terms included: “PIN1” or “rs2233678” in combination with “polymorphism” or “variant” and “cancer” or “neoplasm” or “malignancy”. The reference lists of each comparative study and previous reviews were manually examined to identify additional relevant studies.Studies were selected according to the following inclusion criteria: (1) case-control studies; (2) investigating the association between PIN1 rs2233678 (G>C) polymorphism and cancer risks; (3)cancers diagnosed by histopathology; (4) providing detail genotype frequencies. Studies without detail genotype frequencies were excluded. Titles and abstracts of searching results were screened and full text papers were further evaluated to confirm eligibility. Two reviewers(XH and XZ) independently selected eligible studies. Disagreement between the two reviewers was settled by discussing with the third reviewer (LZ).The following data was collected by two reviewers(XH and XZ) independently using a purpose-designed form: name of first author, publishing time, country where the study was conducted, genotyping methods, ethnicity, cancer types, source of control, number of cases and controls, genotype frequency in cases and controls. Different ethnicity descents were categorized as Asian and Caucasian. Cancer types were classified as breast cancer, squamous cancer , and other cancers . Eligible studies were defined as hospital-based (HB) and population-based (PB) according to the control source.see ), which was modified form a previous meta-analysis The quality of eligible studies was evaluated by three reviewers  independently by scoring according to a “methodological quality assessment scale” (The association strength between −842G>C (rs2233678) polymorphism and cancer risks was measured by odds ratio (OR) with 95% confidence intervals (95% CI). The estimates of pooled ORs were achieved by calculating a weighted average of OR from each study. A 95% CI was used for statistical significance test and a 95% CI without 1 for OR indicating a significantly increased or reduced cancer risk. The pooled ORs were calculated for homozygote comparison (CC versus GG), heterozygote comparison (GC versus GG), dominant (GC/CC versus GG) and recessive (CC versus GC/GG) models, assuming dominant and recessive effects of the variant G allele, respectively. Subgroup analyses were performed according to (i) cancer types, (ii) ethnicities, and (iii) source of control, to examine the impact of these factors on the association. To test the robustness of the association and characterize possible sources of statistical heterogeneity, sensitivity analysis was carried out by excluding studies one-by-one and analyzing the homogeneity and effect size for all of rest studies.2) was used to quantify the degree of heterogeneity between studies and the percentage of τ2 was used to describe the extent of heterogeneity explained Chi-square based Q test was used to check the statistical heterogeneity between studies, and the heterogeneity was considered significant when p<0.10 2 test (P<0.05 means deviated from HWE). All analyses were performed using Stata version 11.0 .HWE (Hardy-Weinberg equilibrium) was tested by Pearson's XThe flow diagram of study identification is shown in heterogeneity<0.01, heterogeneity<0.01, heterogeneity = 0.193) or recessive model OR = 0.653, 95% CI: 0.354, 1.203; Pheterogeneity = 0.088); however, a trend of reduced risk could be drawn.We observed a significantly reduced risk of cancer susceptibility in heterozygote comparison  and dominant model for breast cancer. In the sub-group analyses of squamous cancer, and other cancers, we did find any significant association between −842G/C polymorphisms in the PIN1 promoter region and cancer risk. As for hospital-based studies, we observed a significantly reduced risk of cancer susceptibility in homozygote comparison , heterozygote comparison , dominant model  and recessive model . However, for hospital-based studies, significant association between −842G/C polymorphisms in the PIN1 promoter region and reduced risks of cancers was found only in heterozygote comparison , dominant model . Ethnicity, also, affected cancer susceptibility. In Asians, there was a statistically reduced cancer risk in the comparison of heterozygote  and dominant model . Results for Asians were similar to that of overall comparisons of pooled eligible studies. In Caucasians, however, no significant association was found in each comparison. Taken together, these results revealed that −842G/C polymorphisms in the PIN1 promoter region was only associated with an increased risk of cancer in Asians.We then performed sub-group analyses to investigate the effect of cancer types, ethnicity, and source of control. As for cancer types, increased cancer risk was found in the heterozygote comparison .Heterogeneity between studies in each comparison was shown in Sensitivity analysis was performed to explore individual study's influence on the pooled results by deleting one single study each time from pooled analysis. The results showed that no individual study affected the pooled OR significantly (data not shown), since no substantial change was found.The potential publication bias of the literatures was .evaluated by funnel plot and Egger's test. No visual publication bias was found in the funnel plot . And EggIn the present meta-analysis of 10 case-control studies, including 4619 cancer cases and 4661 controls, a significant association was found between PIN1 −842G/C polymorphism and reduced cancer risk under the heterozygous and dominant genetic models. Under the homozygous and recessive genetic model, there was no significant association between PIN1 −842G/C polymorphism and cancer risk. Overall, a significant association exists between −842G/C polymorphisms in the PIN1 promoter region and cancer risk. This finding indicates that the genetic variant in PIN1 promoter region may crucially modify the susceptibility of cancers.Although PIN1 is not an oncogene itself, it is able to potentiate the function of several oncogenic pathways depending on other oncogenes In stratified analysis by cancer site, we found that −842G/C polymorphism in the PIN1 promoter region was statistically related with reduced breast cancer risks. However, we did not observe any significant association between the genetic variant and the susceptibility of other cancers. However,there are only two studies In the sub-group analysis of ethnicity, we found a significant association between −842G/C polymorphism and reduced risk of cancer in Asians but not in Caucasians. The different cancer risks in Asians and Caucasians were also reported in other meta-analyses During sub-group analyses, we found that control source also affected the association between −842G/C polymorphisms in the PIN1 promoter region and cancer risk. As for hospital-based studies, we observed a significantly reduced risk of cancer susceptibility in homozygote model, and recessive model. However, for hospital-based studies, no significant association between −842G/C polymorphisms in the PIN1 promoter region and risk of cancers was found in homozygote model, and recessive model. Further, it's worth noting that the majority (66.7%) studies of Caucasians use hospital-based controls, while most (57.1%) studies of Asians use population-based controls. So the ethnic interpretations are available to the inconsistency in control source stratification.Some limitations might be included in the meta-analysis. First, we did not search for unpublished studies, so only published studies were included in our meta-analysis. Therefore, publication bias may have occurred although no publication bias was indicated from both visualization of the funnel plot and Egger's test. Second, the sample size of the included studies was not large enough, which could decrease the statistical power to better evaluate the association between −842G/C polymorphism in the PIN1 promoter region and cancer risk. Third, most of the included studies had conducted on Asians, and a few Caucasians. Thus, more samples should be collected from Caucasians.In conclusion, this meta-analysis suggests that the −842G/C polymorphism in PIN1 gene is associated with a significantly reduced risk of cancer, especially in Asian populations. More well-designed studies focusing on different ethnicities and cancer types are warranted in the future.Table S1PRISMA Checklist.(DOC)Click here for additional data file.Table S2Scale for methodological quality assessment.(DOC)Click here for additional data file.


Associations between rs2252004 and BPH risk/aggressiveness were tested using logistic regression. Associations between rs2252004 and clinical parameters including International Prostate Symptom Score (IPSS), total prostate volume (TPV), total PSA (tPSA), and free PSA (fPSA) were evaluated by linear regression. Allele “A” in rs2252004 was significantly associated with increased risk for aggressiveness of BPH in a Chinese population . Patients with the genotype “A/A”  had an increase of IPSS and TPV after treatment . No association was observed between rs2252004, BPH risk, and baseline clinicopathological traits (All P > 0.05). Our study is the first to show that rs2252004 at 10q26 was associated with BPH aggressiveness and efficacy of BPH treatment.Recent studies reported that rs2252004 at 10q26 was significantly associated with prostate cancer (PCa) risk in a Japanese population and was subsequently confirmed in a Chinese population. We aimed to assess the relationship between this locus and risk/aggressiveness of benign prostatic hyperplasia (BPH). The current study included 426 BPH cases and 1,008 controls from Xinhua Hospital in Shanghai, China. All BPH patients were treated with  Benign prostate hyperplasia (BPH) is an independent disease with clinical symptoms similar to those of carcinoma of the prostrate (PCa), and the prevalence of histologically identifiable BPH is >50% for 60-year-old men and ~90% by age 85 years . While sWhile studies have shown that BPH causes significant morbidity, the etiology and determinants of severity of this condition remain poorly understood. According to epidemiological studies, 83.3% of PCa are associated with BPH, and 3–20% of patients who have undergone transurethral prostatectomy (TURP) or open prostatectomy for BPH subsequently develop PCa [Over 40 SNPs have been reported to contribute to PCa risk in different ethnicities . FifteenP = 1.98E − 8) in the Japanese population [A recent GWAS PCa study identified rs2252004 at 10q26 to be significantly associated with PCa risk at a genome-wide significant level  from July 2010 to July 2012. Male volunteers from multiple communities in Shanghai, China, were recruited as controls, from April 2010 to November 2010. All participants gave informed consent, and the study was approved by Xinhua's Ethics Committee prior to involvement in this study.The information we obtained from subjects included the International Prostate Symptom Score (IPSS), quality of life question (IPSS-Q1), postvoid residual volume (PVR), prostate size, serum prostate-specific antigen (PSA) level, liver and renal function, blood glucose level, and routine urine examination.BPH cases included in this study must meet the following criteria: benign prostatic enlargement (BPE) with lower urinary tract symptom (LUTS), age > 45 years, prostate size > 30 mL, IPSS > 7, PVR volume ≤ 1500 mL, and PSA < 4 ng/mL. Patients with PSA ≥ 4 ng/mL were included only after digital rectal examination, true-cut biopsy, and long-time follow-up visit of stabilized PSA in order to make sure without the presence of PCa. Exclusion criteria were history of urinary tract infection (UTI), previous lower tract surgery or procedures, and neurogenic bladder dysfunction. Inclusion criteria for controls included age 40–79 years, clear consciousness, ability to provide a blood sample, and willingness to complete a medical examination. Detailed information of controls was previously reported in Ma et al. [α-adrenergic blockers (doxazosin) and 5 mg of 5α-reductase inhibitors (finasteride) once daily for at least nine months. Patients were classified into “aggressive BPH” group if they suffered from a significant increase in the IPSS score, continuous decrease in maximum urinary flow rate or BPH related complications , or underwent an operation after the combined medication therapy. In contrast, patients with stable disease and without indications to receive invasive treatments were assigned to the nonaggressive group. Thus, 184 aggressive and 242 nonaggressive BPH cases were defined.  Our study population included 426 BPH patients and 1,008 healthy men. BPH cases were treated with a combined therapy of 4 mg  A recent study reported that rs2252004 was associated with PCa risk at a genome-wide significant level in a Japanese population. This finding was confirmed in a Chinese population and evaluated in the current study. The SNP was genotyped for all study subjects using the MassARRAY iPLEX system  at Fudan University in Shanghai, China. Two duplicates and two water samples were included in each 96-well plate as PCR-negative controls. All assays were performed in a blinded fashion. The overall genotyping rate was 98.5%. The average concordance rate between samples was 100% among the duplicated quality control samples.P values were two tailed. An alpha of 0.05 was used to claim statistical significance.  Genotype distributions of the SNP were tested by Hardy-Weinberg equilibrium (HWE). Logistic regression was used to evaluate the association between the SNP and prognosis of BPH (BPH versus controls) and aggressiveness (aggressive versus nonaggressive) with the adjustment of age. The SNP for BPH risk was also estimated using a logistic regression model, assuming an additive mode of inheritance and with the adjustment of age. Clinical variables such as IPSS, TPV, total prostate-free antigen (tPSA), and free prostate-free antigen (fPSA) were not normally distributed and were transformed using log transformations. The changes (after minus before treatment) of IPSS, TPV, tPSA, and fPSA were normally distributed and were not transformed. Association between the SNP and these quantitative clinicopathological traits was analyzed by linear models, adjusting for age. All the above statistical analyses were conducted using PLINK software . ResultsP < 0.05). Therefore, all subsequent statistical analyses were age adjusted. No significant differences in clinical characteristics were found between the aggressive and nonaggressive groups . As shown in P = 0.03). Patients who carry the risk allele “A” of rs2252004 had a 1.42 increased risk for aggressive BPH, compared with patients carrying a “C” allele . Although the frequency of allele “A” was higher in BPH cases (MAF = 0.31) than in controls (MAF = 0.28), the difference in the allele frequency between BPH cases and controls did not reach statistical significance (P = 0.23) (Genotype frequency distributions of rs2252004 were in Hardy-Weinberg Equilibrium (HWE) in both cases and controls (All  = 0.23) .P > 0.05). We did not observe a statistically significant association between genotypes of rs2252004 and basic clinical characteristics, including IPSS, TPV, tPSA, and fPSA . Men who carried the “A/A” genotype  of rs2252004 had an average increase of 1.38 in IPSS, while men carrying “A/C” or “C/C” genotypes had decreased IPSS score of 0.14 and 0.2, respectively . Similarly, men who carried the “A/A” genotype  of rs2252004 had an average increase of 4.62 mL in TPV, while men carrying “AC” or “CC” genotypes had decreased TPV of 0.3 mL and 0.84 mL . This suggested that IPSS and TPV increased after the treatment of men carrying homozygotes for the “A” allele of rs2252004. Significant associations were not observed for rs2252004 with the change of tPSA and fPSA  and 5alpha-reductase inhibitors (5-ARIs) are used for the medical therapy of BPH. The Medical Therapy of Prostatic Symptoms (MTOPS) study reported that during 4.5 years of followup, the progression of BPH, the worsening of symptoms, and BPH-related surgeries decreased more in patients prescribed doxazosin and finasteride combination therapy compared with patients prescribed doxazosin or finasteride monotherapy [BPH is a prevalent disease, especially in old men. This chronic disease has important care implications and financial risks to the health care system. Current management options for BPH include medications and prostate surgery. Taking into account adverse effects of invasive surgery and good therapeutic effect of medical management, more clinical pharmacists and patients tend to select drug therapy as a first treatment choice. At present, alpha-adrenergic receptor blockers  of these genes in a Chinese population [SRD5A2 gene were statistically significantly associated with the change of IPSS and tPSA after treatment, and rs166050 in SRD5A1 was significantly associated with the change of TPV after combined treatment [α-reductase inhibitors and a1-adrenoceptor antagonists.Previous studies have shown that interindividual variation in drug response is partly determined at the genetic level . SRD5A1 pulation . Particureatment . Howeverreatment , 22. TheITGA6 at 2q31, rs339331 in RFX6 at 6q22, and rs103294 in LILRA3, were also significantly associated with BPH risk [We recently observed that several SNPs contributing to PCa risk and identified through GWAS also play an important role in BPH risk and BPH aggressiveness. Three SNPs, rs12621278 in BPH risk . More imThe SNP rs2252004 located in 10q26 was a novel locus recently identified to be associated with PCa risk in a Japanese population and subsequently confirmed in a Chinese population . Our curBRWD2 gene, and rs2252004 was located ~234 kb upstream of the BRWD2 gene. BRWD2 is associated with abnormal pubertal development, such as cryptorchidism, sperm defects and infertility, hypogenitalism, and sparse pubic hair [The nearest gene located on the 10q26 region is the There are two limitations that need to be noted regarding the present study. Firstly, the relatively small sample size led to low statistical power to detect SNP with modest effect size. Secondly, total PSA level of 42% BPH cases exceed the 4 ng/mL. However, all these subjects received transrectal ultrasound guided prostatic true-cut biopsy to rule out PCa and vigilant follow-up observation. Further studies are needed to confirm our results in a larger sample size and with more strict inclusion criteria.Although further elucidation of the biological mechanisms that influence the efficacy of medical treatment for BPH is needed, molecular genetic approaches might provide an opportunity to identify predictive markers of clinical response. In our current study, we successfully identified significant association of rs2252004 in 10q26 and aggressiveness of BPH. This result indicates an important role of rs2252004 in prostatic disease development and future clinical application value.


Mr of approximately 92000, compatible with MMP-9, not observed in db/db mice. These results indicate that higher levels of serine proteinases in plasma may serve as potential markers for kidney changes in db/db mice, whereas a decrease in MMP-9 in the kidney may be related to the glomerular changes.Db/db mice are overweight, dyslipidemic and develop diabetic complications, relevant for similar complications in human type 2 diabetes. We have used db/db and db/+ control mice to investigate alterations in proteinase expression and activity in circulation and kidneys by SDS-PAGE zymography, electron microscopy, immunohistochemistry, Western blotting, and in situ zymography. Plasma from db/db mice contained larger amounts of serine proteinases compared to db/+ mice. Kidneys from the db/db mice had a significantly larger glomerular surface area and somewhat thicker glomerular basement membranes compared to the db/+ mice. Furthermore, kidney extracts from db/+ mice contained metalloproteinases with  One of the most important factors to control in this disease is chronic hyperglycaemia. Several complications, such as retinopathy, peripheral neuropathy, and nephropathy are correlated to elevated HbA control –6. The E control –10. In d control . Matrix  control . MMPs ha control , 14. Bot control . Shifts  control  and an i control . Changes control , 19.  We have recently shown that the levels of MMP-2 and MMP-9 were higher in serum from subjects with type 1 diabetes than in controls, whereas the levels of tissue inhibitors of metalloproteinases (TIMPs) were not affected . It has  The db/db mouse is an animal model for obesity-related diabetes and may be used to study kidney changes in diabetes . These mn = 8) of mixed gender, as well as their nondiabetic heterozygote littermates , were purchased from Harlan  or M&B  and were housed at 23°C and 55% humidity with a 12 : 12 h light-dark cycle. The mice were treated in accordance to the guidelines on accommodation and care of animals formulated by the European Convention for the Protection of Vertebrate Animals for Experimental and Other Scientific Purposes. Blood samples were collected, mixed with heparin, and centrifuged at 11.000 ×g for 10 min at 4°C and plasma isolated. Kidneys were harvested, divided in two and either snap-frozen, fixed in a Zn-based fixative (ZBF) , or fixed in 2% (w/v) glutaraldehyde, 0.5% (w/v) paraformaldehyde in 0.1 mol/l phosphate buffer, pH 7.4. The animal experiments were approved by the Animal Welfare Committee, University of Tromsø.11-12 weeks old db/db diabetic mice  in stacking gel and 7.5% (w/v) in separating gel and electroblotted to a polyvinylidene difluoride membrane . After blockage of nonspecific binding sites with 5% (w/v) non fat milk in Tris-buffered saline, blots were incubated for 1 h at room temperature with antibodies against human MMP-2  and MMP-9 . After washing, the blots were incubated for 1 h at room temperature with horseradish peroxidise- (HRP-) conjugated antibodies MMP-2: Anti-goat , MMP-9: Anti-rabbit  in blockage solution and developed with Western Blot Luminol (Santa Cruz).t-test. P values <0.05 were considered significant.Kidneys were divided in two, and one part was fixed in 2% (w/v) glutaraldehyde , 0.5% (w/v) paraformaldehyde  in 0.1 mol/L phosphate buffer, pH 7.4. Four samples of cortical tissue from each of these half kidneys were cut out, postfixed in 2% (w/v) osmium tetroxide (Agar Scientific), dehydrated in graded alcohol, and embedded in epoxy resin (Agar Scientific). Semithin sections were cut from coded blocks, stained with toluidine blue, and used for light microscopic morphometry using semiautomatic interactive image analysis . Thus, from each of 8 animals, the glomerular surface area was measured for all glomerular profiles of one section from each of 6 independent cortical tissue blocks, and the 6 largest profiles were subjected to further analyses. One block containing central sections of 2 glomeruli was chosen for ultrathin sectioning followed by uranyl acetate/lead citrate contrasting. At medium power electron microscopy, the sections of the glomerular tufts were scanned for thin capillary loops. Five such loops were subjected to high-power microscopy, and the thickness of the basement membrane measured from the 5 thinnest areas of each loop on digital images using semiautomatic interactive image analysis as above. Data from the two animal groups were analyzed for possible statistical differences using student μm thick sections of ZBF-fixed, paraffin-embedded kidneys on Superfrost Plus slides were deparaffinised in xylene and rehydrated in graded alcohol baths. Immunohistochemistry was performed as previously described [+ system-HRP for rabbit primary antibodies, DAKO ). Nuclei were counterstained with Harris haematoxylin (Chemi-Teknikk) and sections mounted with Histokit (Chemi-Teknikk). As negative control, the primary antibody was replaced by 1.5% normal goat serum (Dako) in some sections.Presence and localization of MMP-2 and MMP-9 in kidneys from 4 parallel db/db and db/+ mice was studied by immunohistochemistry using polyclonal antibodies against MMP-2 and MMP-9 . Four-escribed . In brieμm thick sections of ZBF-fixed, paraffin-embedded kidneys were deparaffinised in xylene and rehydrated in graded alcohol baths. 1 mg DQ-gelatin substrate  was dissolved in 1.0 mL Milli-Q water and diluted 1 : 50 in 50 mmol/L Tris-HCl, 150 mmol/L NaCl, 5 mmol/L CaCl2, and 0.2 mmol/L sodium azide (pH 7.6). Sections were incubated with 250 μL of this mixture in a dark humidity chamber at 37°C for 2 h, rinsed, and fixed in 4% (w/v) neutral-buffered formalin. Nuclei were counterstained with DAPI (Invitrogen). To verify the contribution of metallo- and serine proteinases, control slides were preincubated with 20 mmol/L EDTA alone or with 1 mmol/L Pefabloc for 1 h. The inhibitors were also added to the substrate. The level of autofluorescence in the tissue was evaluated by incubating control sections at −20°C for 2 h immediately after the substrate was added. Fluorescence was studied using an Olympus BX51 fluorescence microscope with CellF imaging software .For localization of gelatinolytic activity, in situ zymography was performed as previously described  on the sμL of lysis buffer (0.25% (v/v) Triton X-100 and 10 mmol/L CaCl2 in 0.1 mol/L Hepes, pH 7) per mg tissue (wet weight), using a Tissue Lyser  with 5 mm steal beads at 25 Hz, 2.5 minutes × 2 (4°C) [Unfixed kidneys from 3 parallel db/db and db/+ mice were homogenized with 20  2 (4°C) . The tubr M of approximately 24000, 72000, and 92000, but enzymes withr M above 92000 were also detected. The gelatinase bands atr M 24000 and 92000 were more prominent in plasma from db/db mice than from the control db/+ mice, whereas all other bands were similar in plasma from the two different animal groups. The difference in gelatin-degrading enzymes in samples from db/db and db/+ mice, as determined by SDS-PAGE gelatin zymography using 10% (w/v) polyacrylamide, was further analyzed by using a 7.5% (w/v) polyacrylamide gel to obtain better separation of enzymes withr M between 72000 and 92000. From this experiment  and control mice (db/+) contained gelatinolytic enzymes . The mosperiment , it is ely 92000  were derr M of approximately 45000 (arrow) was observed in all four samples but with the same amounts in diabetic and control mice. To further analyze for the presence of plasminogen activators in plasma from control and diabetic mice, samples were subjected to zymography with gels containing both gelatin and plasminogen substrates. In r M of 80000–850000 and 24000 were not affected, whereas the gelatinase with rM of 72000 was completely abolished , while the enzyme withr M of 24000 and two species withr M of approximately 92000 passed through (F1 and F2). No MMP-9 was detected in the Western blots (not shown). However, MMP-2 was detected in the fractions bound to the column (A1 and A2) as shown in r M of 72000 from both control and diabetes mice detected by gelatin zymography is MMP-2. Some of the gelatin-degrading proteinases found in plasma samples from control and diabetic mice could belong to the MMP family. To test this, we performed Western blotting experiments using antibodies against MMP-2 and MMP-9. In the initial experiments, we could not detect these MMPs with the antibodies used (not shown). Therefore, prior to Western blotting, plasma samples were subjected to gelatin affinity chromatography to remove components that could potentially interfere with Western blotting. Gelatin zymography  showed tP = 0.004). At the ultrastructural level, morphometry was performed to measure basement membrane thickness in glomerular capillary loops. There was a tendency towards thicker basement membranes in glomeruli of db/db mice compared to controls (P = 0.27). The control material contained one outlier (no. 21). If this was excluded, the difference between the two groups was statistically significant (P = 0.033).To document morphological changes in the sections of cortical kidney, tissue from eight db/db and eight db/+ mice were subjected to light microscopic morphometry. The results showed larger glomerular surface areas in sections from db/db mice than in corresponding sections from db+ mice . The difcontrols , althougThe kidney samples were also subjected to immunohistochemistry to test whether MMP-2 and MMP-9 were differently expressed in the db/db and db/+ mice. MMP-2 could be detected both in glomeruli and the tubule systems of kidneys from control and diabetes mice . HoweverTo further analyze for possible presence and activity of gelatin-degrading enzymes in the kidneys, sections from db/db and db/+ mice were subjected to in situ zymography in the absence and presence of EDTA and Pefabloc. Gelatinolytic activity was primarily detected in the tubule system in both control and diabetic mice (rM around 225000), but proteinases migrating as the pro-MMP-9 standard were only present in extracts from control db/+ mice, as shown in r M of 92000 which was of metalloproteinase nature. This enzyme was completely absent in extracts of corresponding tissues from the db/db mice. To further address the question if changes in gelatin-degrading proteinases in plasma in db/db mice are related to similar changes in the kidneys, extracts of kidney tissues were prepared and analyzed by gelatin zymography. Both extracts contained high molecular weight gelatin-degrading proteinases (r M of 92000 from the kidney extract did not bind to the gelatin-Sepharose column but to the lentil lectin column (not shown). Western blotting revealed that at least a part of the enzyme from the db/+ mice extracts withr M of 92000 binding to the lentil lectin column was MMP-9, as can be seen in  To further investigate if this metalloproteinase was MMP-9, kidney extracts were subjected to Western blotting. In the initial experiments, we could not detect MMP-9 (not shown). Extracts were, therefore, subjected to both gelatin-Sepharose and lentil lectin-Sepharose affinity chromatography. MMP-9 is known to bind gelatin through their fibronectin repeats  and to lr M of 92000 compared to extracts from the control mice. In addition, in situ zymography demonstrated that metallo- and serine proteinase activity was mainly found in the tubular system. We also show that the glomerular surface area is larger in db/db mice than in db/+ mice. Our data do also indicate an increased thickness of glomerular basement membranes in db/db mice, in line with other observations from db/db mice [In the present study, we have analyzed gelatinolytic enzymes in plasma and kidney samples from db/db mice and corresponding db/+ control mice. The results presented show that plasma from db/db mice contains increased amounts of gelatin-degrading serine proteinases compared to controls. Further, kidney extracts of db/db mice showed a complete loss of gelatinases with/db mice . The use of well-suited animal models with relevance for human diabetes and kidney complications, in particular, is an important issue. The generation of disease markers, further understanding of pathological mechanisms, testing and generation of new treatments, all depend on good and relevant animal models. The db/db mouse is one such model, in addition to, for example, the ob/ob mouse, the KKAy mouse, the New Zealand obese mouse, and the streptozotocin-induced type 1 diabetes mouse. For studies of kidney changes, the db/db mouse may be a good model system . Rat hasr M of 92000, (consistent with MMP-9) in kidney extracts from db/db mice, which is not reflected in blood samples. Changes in ECM turnover may, therefore, be more prominent in some tissues than others in diabetes. This is an important issue in diabetes research because several organs are affected by the diabetic state. The question is if there is a hyperglycaemia and time dependence of organ affections and to what extent blood markers can be used to predict potential complications in different organs at risk. The data presented show a major decrease in a gelatinase with The data presented here using plasma from db/db mice did not show the same changes in gelatinases as we observed in a previous study on type 1 diabetes in humans. Blood samples from patients with type 1 diabetes contained higher concentrations of MMP-2 and MMP-9 compared to healthy controls . Increasr M of 92000 which was not found in extracts from db/db mice. Western blotting revealed that at least part of this band corresponded to MMP-9. MMP-9 and MMP-2 are known to degrade ECM components such as collagen type IV [r M of 92000 in kidney extracts from db/db mice may contribute to such ECM accumulation. The dynamics of ECM turnover is clearly affected in the kidneys of db/db mice contributing to albuminuria and kidney changes [ The kidney extracts from db/+ control mice contained a metalloproteinase with type IV , 34. Our type IV . The dec changes , as also changes . By in sβ, has been established [β signalling in db/db mice reversed glomerulopathy and mesangial matrix expansion [ In human diabetes, it is well documented that the mesangial matrix is expanded and that the basement membranes of glomeruli are thicker than controls . The molablished . Experimxpansion , 38. Thexpansion , 40.Plasma from db/db mice contained larger amounts of serine proteinases compared to db/+ mice indicating that levels of serine proteinases in plasma may serve as a marker of kidney changes in db/db mice. Furthermore, kidney extracts from db/+ mice contained more MMP-9 than corresponding extracts from db/db mice. This decrease in MMP-9 might contribute to the altered tissue homeostasis in the kidneys of db/db mice, as demonstrated by increased glomerular surface area and thicker glomerular basement membranes, compared to the db/+ mice. There are several issues that need to be addressed in further studies on serine proteinases and MMPs in relation to diabetes and kidney complications. The actual targets for MMPs are poorly defined in kidneys, and the possible importance of MMP changes for diabetes-related pathogenesis and ECM alterations has so far not been addressed in any detail. There is a potential to use MMPs and related proteinases in plasma as markers for kidney changes. MMPs and TIMPs may also be potential targets for therapeutic interventions. The db/db mouse model is useful for studying kidney changes in diabetes , and our


To describe the prevalence and associations of posterior vitreous detachment (PVD) in a rural adult Chinese population.All eligible subjects were requested to carry out a comprehensive eye examination; PVD was a pre-specified outcome variable and was determined via biomicroscopical examination (slit-lamp biomicroscopy) with a +90-D preset lens after mydriasis. Prevalence was standardized to China population census (2000).5890 (86.2%) subjects completed the examination of slit-lamp biomicroscopy with a +90-D lens. PVD was present in 160 participants (2.7%); the standardized prevalence was 2.0% . PVD developed increasingly with age (P for trend < 0.001) for both men and women. Using a multivariate regression model, older people were found to run a higher risk of developing PVD than younger people, and women were found to have a higher risk than men . Diabetes, hypertension, smoking, drinking, and intraocular pressure (IOP) were not significantly associated with PVD.About one in fifty people is found to have PVD in this population-based study. Age and female are independently associated with PVD occurrence. The vitreous body plays important roles in both intraocular metabolic exchange and the stability of the retina. Losing its gel-like consistency with aging, the vitreous body may eventually detach from the retina and optic disc, clinically described as posterior detachment of the vitreous body (PVD) . When PVHowever, information on the prevalence of PVD is largely lacking. The limited data to date have mostly come from hospital-based and post-mortem studies ,3,8-10. The Handan Eye Study is a population-based study designed to determine the prevalence of blindness, visual impairment, and common eye diseases in a rural population of northern China. Details of the study design, sampling plan, and baseline data are reported elsewhere -16. EthiIn brief, a total of 6830 Han Chinese aged 30 years and older were randomly selected from 13 villages in Yongnian County, Handan City and were invited to participate in the Handan Eye Study between October 2006 and October 2007. 80% of the population work as peasants, and 98% are Hans. Socioeconomically, they are at the average level in mainland China .All eligible individuals underwent a comprehensive eye examination after pupil dilation, including standardized refraction and biomicroscopical examination (slit-lamp biomicroscopy) with a +90-D preset lens by a trained ophthalmologist.In this study, a senior retina specialist (DXR) would diagnose the vitreous conditions by observing kinetic movement of the posterior vitreous during saccadic movement of the eyes via a slit-lamp biomicroscopy with a +90-D preset lens. PVD was defined as separation of the posterior vitreous cortex from the retinal surface with a peripapillary ring and/or an optically empty subhyaloid space ,19. The Refractive error was defined as spherical equivalent (sphere + cylinder/2) . High myDiabetes mellitus was diagnosed from self-reported history of medication and/or a fasting plasma glucose ≥7.0 mM (126 mg/dL). Hypertension was defined as having a history of hypertension on medication or elevated blood pressure when measured in the examination (diastolic blood pressure ≥90 mmHg or systolic blood pressure ≥140 mmHg). Smoking referred to 1 to 100 cigarettes per day and continued to 100 cigarettes. Drinking referred to often drinking (at least one time per week at least 50 g per time).Statistical analysis was performed by using a statistical software package (SAS9.1.3). All statistical tests were significant if P values were less than 0.05. The association with PVD, such as gender, intraocular hypertension and health was estimated by the odds ratio (OR) and its 95% confidence interval (95% CI) with multivariate logistic regression model.Of the 6830 participants, 5890 (86.2%) completed the examination of slit-lamp biomicroscopy with a +90-D preset lens in the county hospital. 807 (11.8%) at temporary study sites in the villages and 114 (1.7%) at home completed the preset lens examination without mydriasis, which might have made it impossible to detect PVD, and, therefore, we excluded these participants from the calculation of prevalence. An additional 19 (0.28%) people had no PVD data because of media opacities or missing data. The mean age of the participants included in the analysis was 52.0 ± 11.5 years and 54.8% were female. Those who did not complete the examination were older, having higher income, with less education, and male gender and were more likely to have no health insurance.PVD was present in 160 participants (2.7%). Standardized directly by age and gender according to the national population census (China 2000), the prevalence of PVD was 2.0% . PVD was present only in 3 participants who were 30+ years old and 40+ years old. PVD was detected in 53 (53/2660 = 1.99%) male subjects and 107 (107/3230 = 3.31%) in female subjects. PVD developed increasingly with age (P for trend < 0.001) for both men and women ] (6.6%) than in subjects with moderate myopia  2.3%) and those with mild myopia and hyperopia  (2.6%). However, the trend of increasing prevalence with myopic SE was higher only in 30~39-year-old group (P for trend = 0.005), but not in other age groups (P > 0.05). The prevalence of PVD even decreased with the increase of myopic SE in 70 + -year-old group (P for trend =0.023). In high myopia group, age was not associated with prevalence of PVD (p for trend = 0.45), while in mild and moderate groups and hyperopia group, prevalence of PVD increased remarkably with age (p for trend < 0.001). . Foos . Chuo etIn our study, those with high myopia had higher prevalence (6.6%) compared with normal people (2.7%). High myopia seems to increase the risk of PVD in younger people but not in elderly and high myopia was not found to be associated with PVD in multivariate logistic regression models. This was inconsistent with previous hospital-based studies ,8,12. ThThough high myopia was not observed to be associated with the prevalence of PVD in this study, the age of onset of PVD might be earlier in eyes with high myopia than in lower myopic eyes or normal eyes Figure . And in No association was found between PVD and diabetics, hypertension, smoking, drinking or IOP. Foos et al., after comparing the prevalence of PVD in their autopsy study between diabetic subjects and the general population, found that PVD was significantly more common in the eyes of diabetics including those with no retinopathy . HoweverOur study has limitations. Although PVD was a pre-specified outcome variable in this study and based on a large population and retinal examination was conducted by an experienced ophthalmologist. We may have underestimated the prevalence: 1) The diagnosis of PVD was examiner-dependent without objective verifications like B scan or OCT examinations, although a prospective study showed that ultrasonography and slit-lamp biomicroscopy were the most reliable noninvasive clinical techniques to detect PVD prior to vitrectomy . In thisIn conclusion, about one in fifty people were found to have PVD in this adult Chinese population-based study. PVD, as a pre-specified outcome variable, was diagnosed on the basis of slit-lamp biomicroscopy with a +90-D preset lens after mydriasis alone without OCT or B-scan. Age and female were independently associated with PVD occurrence, while diabetics, hypertension, smoking, drinking, and IOP were not associated with PVD. High myopia was found to increase the risk of PVD in younger people but not in elderly.The authors declare that they have no competing interests.NLW participated in the design of the study, drafting the article, obtaining funding and supervision. ZJS, YBL participated in providing conception and design, drafting the article, data analysis and interpretation. XRD, FHW, SZL carried out the eye examination and participated in data acquisition, providing conception and design. YP, DTL participated in data analysis, revising the article. XYP participated in providing conception and design, data interpretation, revising the article. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2415/13/33/prepub