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1.  Repeatability and Reproducibility of a Double-Pass Optical Quality Analysis Device 
PLoS ONE  2015;10(2):e0117587.
Purpose
To evaluate the repeatability and reproducibility of a double-pass instrument (OQASII, Visiomereics SL, Spain), which objectively measures overall optical quality of the human eyes.
Methods
The right eye of 119 healthy subjects with best corrected visual acuity of 20/25 or better was included in this prospective, comparative, observational study. Two separate tests with OQASII were conducted sequentially on the same day by two different examiners. A week later, the first examiner conducted the third measurement. All subjects underwent three consecutive tests during each session. The repeatability and reproducibility of the modulation transfer function cut off frequency (MTF cutoff), the Strehl ratio, the OQAS values (OVs) at contrasts of 100%, 20% and 9%, and the objective scatter index (OSI) were analyzed.
Results
For MTF cutoff, Strehl ratio, OV100%, OV20%, OV9%, and OSI, the mean values were 39.32±9.75cpd, 0.22±0.06, 1.31±0.33, 1.33±0.39, 1.33±0.41, 0.60±0.42, respectively. Repeatability and reproducibility were good with a very low coefficient of variation and high interclass correlation coefficients (>0.88) for all parameters. Bland-Altman plots showed good correlation with 95% limits of agreement ranged from -6.04 to 6.78cpd, -0.05 to 0.05, -0.20 to 0.23, -0.29 to 0.32, -0.40 to 0.42, -0.23 to 0.21 in inter-observer, and -6.56 to 7.42cpd, -0.06 to 0.06, -0.22 to 0.24, -0.30 to 0.32, -0.35 to 0.34, -0.24 to 0.23 in inter-visit, respectively.
Conclusion
The OQASII system yields excellent repeatability and good reproducibility for objective measurements of overall optical quality in clinic.
doi:10.1371/journal.pone.0117587
PMCID: PMC4315520  PMID: 25646846
2.  A novel method for detection of HBVcccDNA in hepatocytes using rolling circle amplification combined with in situ PCR 
BMC Infectious Diseases  2014;14(1):608.
Background
Intrahepatic hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) is the original template for HBV replication. The persistence of cccDNA is responsible for the recurrence of HBV infection. The detection of cccDNA can help the development of new antiviral drugs against HBV replication links, and reduce the resistance and recurrence as well as to discover extrahepatic HBV infection. In situ polymerase chain reaction (IS-PCR) can be used to determine the distribution and localization of cccDNA in liver tissues, but it is hampered by its low sensitivity and specificity. We developed a novel method to detect HBV cccDNA using rolling circle amplification (RCA) combined with IS-PCR.
Methods
Biopsy liver tissues were obtained from 26 patients with HBV infection, including 10 chronic hepatitis B (CHB), 6 liver cirrhosis (LC) and 10 hepatocellular carcinoma (HCC) patients. Four pairs of primers were designed to mediating RCA for the first round amplification of HBV cccDNA specifically. The liver tissue sections from patients were treated by plasmid-safe ATP-dependent DNase (PSAD) prior to RCA. After RCA, HBV cccDNA was further amplified by a pair of selective primers labeled digoxigenin that target the gap region between the two direct repeat regions (DR1 and DR2) of the virus via IS-PCR.
Results
HBVcccDNA was expressed and located in hepatocyte nucleus in 19 patients (73.07%). Compared with the IS-PCR, the introduction of RCA increase the limit of detection. RCA combined with IS-PCR yielded strong positive signals in HCC liver tissue in spite of low copy number cccDNA (2 copies of target sequence per cell), meanwhile, no positive signal was detected via negative control.
Conclusions
RCA combined with IS-PCR is an effective and practicable method which could detect the presence of low copy number of cccDNA sensitively and specifically, and reflect the relationship between cccDNA expression level and liver tissue pathological characteristics.
doi:10.1186/s12879-014-0608-y
PMCID: PMC4264245  PMID: 25465805
Covalently closed circular DNA; Hepatitis B virus; In situ PCR; Rolling circle amplification
3.  Superoxide Production by a Manganese-Oxidizing Bacterium Facilitates Iodide Oxidation 
The release of radioactive iodine (i.e., iodine-129 and iodine-131) from nuclear reprocessing facilities is a potential threat to human health. The fate and transport of iodine are determined primarily by its redox status, but processes that affect iodine oxidation states in the environment are poorly characterized. Given the difficulty in removing electrons from iodide (I−), naturally occurring iodide oxidation processes require strong oxidants, such as Mn oxides or microbial enzymes. In this study, we examine iodide oxidation by a marine bacterium, Roseobacter sp. AzwK-3b, which promotes Mn(II) oxidation by catalyzing the production of extracellular superoxide (O2−). In the absence of Mn2+, Roseobacter sp. AzwK-3b cultures oxidized ∼90% of the provided iodide (10 μM) within 6 days, whereas in the presence of Mn(II), iodide oxidation occurred only after Mn(IV) formation ceased. Iodide oxidation was not observed during incubations in spent medium or with whole cells under anaerobic conditions or following heat treatment (boiling). Furthermore, iodide oxidation was significantly inhibited in the presence of superoxide dismutase and diphenylene iodonium (a general inhibitor of NADH oxidoreductases). In contrast, the addition of exogenous NADH enhanced iodide oxidation. Taken together, the results indicate that iodide oxidation was mediated primarily by extracellular superoxide generated by Roseobacter sp. AzwK-3b and not by the Mn oxides formed by this organism. Considering that extracellular superoxide formation is a widespread phenomenon among marine and terrestrial bacteria, this could represent an important pathway for iodide oxidation in some environments.
doi:10.1128/AEM.00400-14
PMCID: PMC3993295  PMID: 24561582
4.  Microbiota of the seminal fluid from healthy and infertile men 
Fertility and sterility  2013;100(5):1261-1269.
Objective:
To explore potential causes of male infertility by determining the composition and structure of commensal bacterial communities in seminal fluids.
Design:
Microscopy of gram stained semen samples and classification of 16S rRNA gene sequences to determine the species composition of semen bacterial communities.
Setting(s):
Clinical andrology laboratory and academic research laboratories.
Patient(s):
19 sperm donors and 58 infertility patients.
Intervention(s):
None.
Main Outcome Measure(s):
Classification of 16S rRNA gene sequences, clustering of seminal microbial communities, and multiple statistical tests.
Result(s):
High numbers of diverse kinds of bacteria were present in most samples of both sperm donors and infertility patients. The bacterial communities varied widely between subjects, but they could be clustered into six groups based on similarities in composition and the rank abundances of taxa. Overall there were no significant differences between sperm donors and infertility patients. However, multiple statistical tests showed a significant negative association between sperm quality and the presence of Anaerococcus. The results also indicated that many of the bacterial taxa identified in semen also occur in the vaginal communities of some women, especially those with bacterial vaginosis, which suggests heterosexual sex partners may share bacteria.
Conclusion(s):
Diverse kinds of bacteria were present in the human semen, there were no significant differences between sperm donors and infertility patients, The presence of Anaerococcus might be a biomarker for low sperm quality.
doi:10.1016/j.fertnstert.2013.07.1991
PMCID: PMC3888793  PMID: 23993888
semen; microbiome; bacterial communities; fertility; reproduction
5.  Biogenesis of C-Glycosyl Flavones and Profiling of Flavonoid Glycosides in Lotus (Nelumbo nucifera) 
PLoS ONE  2014;9(10):e108860.
Flavonoids in nine tissues of Nelumbo nucifera Gaertner were identified and quantified by high-performance liquid chromatography with diode array detector (HPLC-DAD) and HPLC-electrospray ionization-mass spectrometry (HPLC-ESI-MSn). Thirty-eight flavonoids were identified; eleven C-glycosides and five O-glycosides were discovered for the first time in N. nucifera. Most importantly, the C-glycosyl apigenin or luteolin detected in lotus plumules proved valuable for deep elucidation of flavonoid composition in lotus tissues and for further utilization as functional tea and medicine materials. Lotus leaves possessed the significantly highest amount of flavonoids (2.06E3±0.08 mg 100 g−1 FW) and separating and purifying the bioactive compound, quercetin 3-O-glucuronide, from leaves showed great potential. In contrast, flavonoids in flower stalks, seed coats and kernels were extremely low. Simultaneously, the optimal picking time was confirmed by comparing the compound contents in five developmental phases. Finally, we proposed the putative flavonoid biosynthesis pathway in N. nucifera.
doi:10.1371/journal.pone.0108860
PMCID: PMC4184820  PMID: 25279809
6.  Diagnostic application of PIK3CA mutation analysis in Chinese esophageal cancer patients 
Diagnostic Pathology  2014;9(1):153.
Background
The PIK3CA gene mutation was found to associate with prognosis and might affect molecular targeted therapy in esophageal carcinoma (EC). The aim of this study is to compare different methods for analyzing the PIK3CA gene mutation in EC.
Methods
Genomic DNA was extracted from 106 surgically resected EC patient tissues. The PIK3CA mutation status (exons 9 and 20) were screened by mutant-enrich liquid chip (ME-Liquidchip), Sanger sequencing, and pyrosequencing. And all samples with mutations were independently reassessed using amplification refractory mutation system (ARMS) methods again.
Results
PIK3CA mutation rates were identified as 11.3% (12/106) by ME-Liquidchip. 10 mutations occurred in exon 9 and 2 in exon 20, including G1624A:E542K (n = 4), G1633A:E545K (n = 6) and A3140G:H1047R (n = 2). The results were further verified by ARMS methods. Among these 12 cases characterized for PIK3CA mutation, however, only 7 and 6 cases were identified by Sanger sequencing (6.6%,7/106) and pyrosequencing (5.7%,6/106), respectively.
Conclusion
Sanger sequencing and pyrosequencing are less sensitive and are not efficiently applicable to the detection of PIK3CA mutation in EC samples. Choosing between ME-Liquidchip and ARMS will depend on laboratory facilities and expertise.
Virtual Slides
The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/13000_2014_153
doi:10.1186/s13000-014-0153-4
PMCID: PMC4149237  PMID: 25106743
Esophageal carcinoma; Mutation; PIK3CA gene
7.  Long Noncoding RNA-EBIC Promotes Tumor Cell Invasion by Binding to EZH2 and Repressing E-Cadherin in Cervical Cancer 
PLoS ONE  2014;9(7):e100340.
In recent years, long noncoding RNAs (lncRNAs) have been demonstrated to play key roles in tumorgenesis. However, the contributions of lncRNAs to cervical cancer (CC) remain largely unknown. In this study, differentially expressed lncRNAs and mRNAs in cervical cancer and paired peritumoral tissues were detected by transcriptome microarray analysis. We found 708 probe sets of lncRNAs increased and 836 probe sets decreased in CC tissues, while 1288 mRNA differential probe sets increased and 901 mRNA probe sets decreased. The results were validated by quantitative real-time polymerase chain reaction (qPCR). Then, we found a specific differentially expressed lncRNA can physically bind to enhancer of zeste homolog2 (EZH2) by using RNA immunoprecipitation. We termed it as EZH2-binding lncRNA in cervical cancer [lncRNA-EBIC]. Wound healing assays and Matrigel invasion assays were used to determine the function of this lncRNA by silencing it. We observed that the migration and invasion of cervical cancer cells in vitro were inhibited upon suppression of lncRNA-EBIC by siRNA. We also found that the association between lncRNA-EBIC and EZH2 was required for the repression of E-cadherin, which was a key molecular in the metastasis of cervical cancer.
Conclusion
These results demonstrated that lncRNA-EBIC was an oncogenic lncRNA, which could promote tumor cell invasion in CC by binding to EZH2 and inhibiting E-cadherin expression.
doi:10.1371/journal.pone.0100340
PMCID: PMC4090119  PMID: 25007342
8.  Effects of postmastectomy radiotherapy on prognosis in different tumor stages of breast cancer patients with positive axillary lymph nodes 
Cancer Biology & Medicine  2014;11(2):123-129.
Objective
To explore the effects of postmastectomy radiotherapy (PMRT) on the locoregional failure-free survival (LRFFS) and overall survival (OS) of breast cancer patients under different tumor stages and with one to three positive axillary lymph nodes (ALNs).
Methods
We conducted a retrospective review of 527 patients with one to three positive lymph nodes who underwent modified radical or partial mastectomy and axillary dissection from January 2000 to December 2002. The patients were divided into the T1-T2 N1 and T3-T4 N1 groups. The effects of PMRT on the LRFFS and OS of these two patient groups were analyzed using SPSS 19.0, Pearson’s χ2-test, Kaplan-Meier method, and Cox proportional hazard model.
Results
For T1-T2 N1 patients, no statistical significance was observed in the effects of PMRT on LRFFS [hazard ratio (HR)=0.726; 95% confidence interval (CI): 0.233-2.265; P=0.582] and OS (HR=0.914; 95% CI: 0.478-1.745; P=0.784) of the general patients. Extracapsular extension (ECE) and high histological grade were the risk factors for LRFFS and OS with statistical significance in multivariate analysis. Stratification analysis showed that PMRT statistically improved the clinical outcomes in high-risk patients [ECE (+), LRFFS: P=0.026, OS: P=0.007; histological grade III, LRFFS: P<0.001, OS: P=0.007] but not in low-risk patients [ECE (–), LRFFS: P=0.987, OS: P=0.502; histological grade I-II, LRFFS: P=0.816, OS: P=0.296]. For T3-T4 N1 patients, PMRT effectively improved the local control (HR=0.089; 95% CI: 0.210-0.378; P=0.001) of the general patients, whereas no statistical effect was observed on OS (HR=1.251; 95% CI: 0.597-2.622; P=0.552). Absence of estrogen receptors and progesterone receptors (ER/PR) (–) was an independent risk factor. Further stratification analysis indicated a statistical difference in LRFFS and OS between the high-risk patients with ER/PR (–) receiving PMRT and not receiving PMRT [ER/PR (–), LRFFS: P=0.046, OS: P=0.039]. However, PMRT had a beneficial effect on the reduction of locoregional recurrence (LRR) but not in total mortality [ER/PR (+), LRFFS: P<0.001, OS: P= 0.695] in T3-T4 N1 patients with ER/PR (+) who received endocrine therapy.
Conclusion
PMRT could reduce ECE (+), histological grade III-related LRR, and total mortality of T1-T2 N1 patients. T3-T4 N1 patients with ER/PR (–) could benefit from PMRT by improving LRFFS and OS. However, PMRT could only reduce LRR but failed to improve OS for T3-T4 N1 patients with ER/PR (+) who received endocrine therapy.
doi:10.7497/j.issn.2095-3941.2014.02.007
PMCID: PMC4069797  PMID: 25009754
Breast cancer; positive lymph nodes; postmastectomy radiotherapy (PMRT); locoregional failure-free survival (LRFFS); overall survival (OS)
9.  Bacterial Cellulose Membranes Used as Artificial Substitutes for Dural Defection in Rabbits 
To improve the efficacy and safety of dural repair in neurosurgical procedures, a new dural material derived from bacterial cellulose (BC) was evaluated in a rabbit model with dural defects. We prepared artificial dura mater using bacterial cellulose which was incubated and fermented from Acetobacter xylinum. The dural defects of the rabbit model were repaired with BC membranes. All surgeries were performed under sodium pentobarbital anesthesia, and all efforts were made to minimize suffering. All animals were humanely euthanized by intravenous injection of phenobarbitone, at each time point, after the operation. Then, the histocompatibility and inflammatory effects of BC were examined by histological examination, real-time fluorescent quantitative polymerase chain reaction (PCR) and Western Blot. BC membranes evenly covered the surface of brain without adhesion. There were seldom inflammatory cells surrounding the membrane during the early postoperative period. The expression of inflammatory cytokines IL-1β, IL-6 and TNF-α as well as iNOS and COX-2 were lower in the BC group compared to the control group at 7, 14 and 21 days after implantation. BC can repair dural defects in rabbit and has a decreased inflammatory response compared to traditional materials. However, the long-term effects need to be validated in larger animals.
doi:10.3390/ijms150610855
PMCID: PMC4100185  PMID: 24937688
bacterial cellulose; artificial dura mater; histocompatibility; inflammatory cytokines
10.  Reference Gene Selection for Quantitative Real-Time RT-PCR Normalization in Iris. lactea var. chinensis Roots under Cadmium, Lead, and Salt Stress Conditions 
The Scientific World Journal  2014;2014:532713.
Quantitative real time PCR (RT-qPCR) has emerged as an accurate and sensitive method to measure the gene expression. However, obtaining reliable result depends on the selection of reference genes which normalize differences among samples. In this study, we assessed the expression stability of seven reference genes, namely, ubiquitin-protein ligase UBC9 (UBC), tubulin alpha-5 (TUBLIN), eukaryotic translation initiation factor (EIF-5A), translation elongation factor EF1A (EF1α), translation elongation factor EF1B (EF1b), actin11 (ACTIN), and histone H3 (HIS), in Iris. lactea var. chinensis (I. lactea var. chinensis) root when the plants were subjected to cadmium (Cd), lead (Pb), and salt stress conditions. All seven reference genes showed a relatively wide range of threshold cycles (Ct) values in different samples. GeNorm and NormFinder algorithms were used to assess the suitable reference genes. The results from the two software units showed that EIF-5A and UBC were the most stable reference genes across all of the tested samples, while TUBLIN was unsuitable as internal controls. I. lactea var. chinensis is tolerant to Cd, Pb, and salt. Our results will benefit future research on gene expression in response to the three abiotic stresses.
doi:10.1155/2014/532713
PMCID: PMC4058273  PMID: 24977206
11.  Association between mammographic features and clinicopathological characteristics in invasive ductal carcinoma of breast cancer 
Molecular and Clinical Oncology  2014;2(4):623-629.
The aim of the present study was to evaluate the association between mammographic features and clinicopathological characteristics in invasive ductal carcinoma. A total of 231 patients were retrospectively reviewed from January, 2011 to December, 2012. Statistical analysis was performed using Fisher’s exact test, χ2 test, Spearman’s correlation and logistic regression, as appropriate. Of the 231 patients who underwent mammography, malignant calcifications were significantly more frequent in carcinomas that were human epidermal growth factor receptor 2 (HER2)-positive (P=0.001) or had a >2 cm size tumor (P=0.006). The pleomorphic-type was correlated with a p53-positive status (P=0.039) or lymph node metastasis (P=0.048), whereas the indistinct amorphous-type was associated with a HER2-positive status (P=0.026). An evident mass was frequently observed in higher Ki-67 expression-level tumors (P=0.002). In conclusion, the aforementioned correlations are noteworthy as they potentially reflect tumor attributes and may serve as a guide for treatment.
doi:10.3892/mco.2014.297
PMCID: PMC4051570  PMID: 24940507
mammography; breast neoplasm; clinic; invasive ductal carcinoma; pathology
12.  Bactericidal/permeability-increasing protein originates in both the testis and the epididymis and localizes in mouse spermatozoa 
Asian Journal of Andrology  2014;16(2):309-313.
Bactericidal/permeability–increasing protein (BPI) is an endogenous antibiotic protein with activity against gram–negative bacteria. In the present study, we examined the expression of BPI in postnatal mouse testes and epididymides as well as the subcellular localization within epididymal spermatozoa. Our results showed that, BPI mRNA was expressed in testis and epididymis independently. Throughout the epididymis, the BPI protein level gradually decreased in the epididymal epithelium in a spatial manner, specialized within the cytoplasm of clear cells in the cauda part. We detected BPI proteins in intact acrosome, implying its testicular origin; on the other hand, after the acrosome reaction, BPI proteins were observed dispersed across the entire sperm head, especially enriched at the equatorial segment. Our findings suggested a dual origin of the BPI that generated both in the testis and epididymis, and associated with mouse spermatozoa. BPI protein might be involved in the dynamics modification of the sperm plasma membrane and also the fertilization process.
doi:10.4103/1008-682X.122583
PMCID: PMC3955346  PMID: 24457839
acrosome; antimicrobial protein; bactericidal/permeability-increasing protein; epididymis; testis
13.  A New Corrosion Sensor to Determine the Start and Development of Embedded Rebar Corrosion Process at Coastal Concrete 
Sensors (Basel, Switzerland)  2013;13(10):13258-13275.
The corrosion of reinforcements induced by chloride has resulted to be one of the most frequent causes of their premature damage. Most corrosion sensors were designed to monitor corrosion state in concrete, such as Anode-Ladder-System and Corrowatch System, which are widely used to monitor chloride ingress in marine concrete. However, the monitoring principle of these corrosion sensors is based on the macro-cell test method, so erroneous information may be obtained, especially from concrete under drying or saturated conditions due to concrete resistance taking control in macro-cell corrosion. In this paper, a fast weak polarization method to test corrosion state of reinforcements based on electrochemical polarization dynamics was proposed. Furthermore, a new corrosion sensor for monitoring the corrosion state of concrete cover was developed based on the proposed test method. The sensor was tested in cement mortar, with dry-wet cycle tests to accelerate the chloride ingress rate. The results show that the corrosion sensor can effectively monitor chloride penetration into concrete with little influence of the relative humidity in the concrete. With a reasonable corrosion sensor electrode arrangement, it seems the Ohm-drop effect measured by EIS can be ignored, which makes the tested electrochemical parameters more accurate.
doi:10.3390/s131013258
PMCID: PMC3859062  PMID: 24084117
corrosion sensor; cement mortar resistance; corrosion rate; reference electrode; EIS
14.  The relationship between Lin28 and the chemotherapy response of gastric cancer 
OncoTargets and therapy  2013;6:1341-1345.
Objective
The aim of the study reported here was to identify whether a stem cell biomarker, Lin28, may predict the pathologic tumor response to neoadjuvant chemotherapy for patients with locally advanced gastric cancer.
Methods
The study enrolled 47 patients with gastric cancer who underwent neoadjuvant chemotherapy followed by surgery between July 2004 and March 2012. Cancer tissue was biopsied by gastroscopy and Lin28 expression in the tissue was measured by immunohistochemistry. Statistical analyses were performed to identify the relationship between Lin28 expression and tumor regression grade.
Results
Of the 47 cases, pathologic nonresponse was observed in 29 (61.7%) and pathologic response in 18 (38.3%). Receiver-operating characteristic curve analysis showed that the histoscore of Lin28 expression with 0.325 as a cutoff value could differentiate between pathologic response and nonresponse. Multivariable analysis showed that Lin28 expression was an independent predictive factor for pathologic response to neoadjuvant chemotherapy (P = 0.006).
Conclusion
Lin28 expression was associated with pathologic tumor response in locally advanced gastric cancer patients undergoing neoadjuvant chemotherapy. This may suggest that Lin28 can serve as a predictive biomarker for neoadjuvant chemotherapy in patients with gastric cancer.
doi:10.2147/OTT.S45705
PMCID: PMC3789745  PMID: 24098084
neoadjuvant chemotherapy; pathologic tumor response; stem cell biomarker; immunohistochemistry
15.  Analytical solution for light propagation in a two-layer tissue structure with a tilted interface for breast imaging 
Applied optics  2006;45(20):5027-5036.
Reflectance measurement of breast tissue is influenced by the underlying chest wall, which is often tilted as seen by the detection probe. We develop an analytical solution of light propagation in a two-layer tissue structure with tilted interface and refractive index difference between the layers. We validate the analytical solution with Monte Carlo simulations and phantom experiments, and a good agreement is seen. The influence of varying the tilting angle of the interface on the reflectance is discussed for two types of layered structures. Further, we apply the developed analytical solution to obtain the optical properties of breast tissue and chest wall from clinical data. Inverse calculation using the developed solution applied to the data obtained from Monte Carlo simulations shows that the optical properties of both layers are obtained with higher accuracy as compared to using a simple two-layer model ignoring the interface tilt. This is expected to improve the accuracy in estimating the optical properties of breast tissue, thus enhancing the accuracy of optical tomography of breast tumors.
PMCID: PMC3776600  PMID: 16807614
16.  Gallbladder Gallstone Disease Is Associated with Newly Diagnosed Coronary Artery Atherosclerotic Disease: A Cross-Sectional Study 
PLoS ONE  2013;8(9):e75400.
Background and Aims
The association between gallstone disease and coronary artery atherosclerotic disease (CAD) remains unclear. To clarify their relationship, patients with CAD newly diagnosed by coronary angiography were investigated in this cross-sectional study.
Methods
The study cohort consisted of 1,270 patients undergoing coronary angiography for the first time between January 2007 and September 2011. Patients with ≥50% diameter stenosis in any major coronary artery on coronary angiography were defined as being CAD positive (n = 766) and those with no stenosis as CAD negative (n = 504). Multivariate logistic regression was used to investigate the relationship between gallstone disease and CAD. The odds ratios (OR) of factors associated with CAD were calculated. In addition, CAD-positive and CAD-negative patients were matched one-to-one by age, gender and metabolic syndrome (MetS), and the association between gallbladder disease and CAD was determined.
Results
The prevalence of gallstone disease was significantly higher in CAD-positive than in CAD negative patients (149/766 [19.5%] vs 57/504 [11.3%], P<0.01). Gallstone disease was significantly associated with CAD (adjusted OR = 1.59, 95% confidence interval [CI] 1.10–2.31). Following matched pairing of 320 patients per group, gallstone disease remained significantly associated with CAD (adjusted OR = 1.69, 95% CI: 1.08–2.65).
Conclusion
Gallstone disease is strongly associated with CAD diagnosed by coronary angiography.
doi:10.1371/journal.pone.0075400
PMCID: PMC3776774  PMID: 24058685
17.  Electrochemically hydrogenated TiO2 nanotubes with improved photoelectrochemical water splitting performance 
Nanoscale Research Letters  2013;8(1):391.
One-dimensional anodic titanium oxide (ATO) nanotube arrays hold great potential as photoanode for photoelectrochemical (PEC) water splitting. In this work, we report a facile and eco-friendly electrochemical hydrogenation method to modify the electronic and PEC properties of ATO nanotube films. The hydrogenated ATO (ATO-H) electrodes present a significantly improved photocurrent of 0.65 mA/cm2 in comparison with that of pristine ATO nanotubes (0.29 mA/cm2) recorded under air mass 1.5 global illumination. The incident photon-to-current efficiency measurement suggests that the enhanced photocurrent of ATO-H nanotubes is mainly ascribed to the improved photoactivity in the UV region. We propose that the electrochemical hydrogenation induced surface oxygen vacancies contribute to the substantially enhanced electrical conductivity and photoactivity.
doi:10.1186/1556-276X-8-391
PMCID: PMC3973816  PMID: 24047205
TiO2 nanotubes; Electrochemical reductive doping; Hydrogenation; Photoelectrochemical water splitting
18.  A novel subtype of primary prostatic adenocarcinoma: A case report 
Oncology Letters  2013;6(5):1303-1306.
The present study reports the novel case of an 81-year-old male with prostatic adenocarcinoma (PAC), whose histopathological study revealed a pure urothelial carcinoma (UC) that originated, however, from the prostatic glandular epithelium. The levels of serum prostate-specific antigen (PSA) were extraordinarily high in this patient. An MRI scan indicated a prostatic neoplasm and no malignant changes were observed in the bladder or other areas of the urinary tract. Hematoxylin and eosin-stained sections revealed a diagnosis of pure UC with no other form of differentiation (typical adenocarcinoma or squamous differentiation). The immunohistochemical findings were positive for PSA and P504S, and negative for CK7, CK20, 34βE12 and p63. A diagnosis of primary PAC (solid carcinoma) originating from the prostate was made based on the clinical, histopathological and immunohistochemical observations. This case was susceptible to diagnostic errors, however, a novel subtype of primary PAC was identified and termed the UC-like subtype.
doi:10.3892/ol.2013.1557
PMCID: PMC3813718  PMID: 24179513
adenocarcinoma of the prostate; histopathological feature; urothelial carcinoma
19.  Subsequent intra-abdominal fibromatosis mimicking recurrent gastrointestinal stromal tumor 
Diagnostic Pathology  2013;8:125.
Abstract
Intra-abdominal fibromatosis (IAF) commonly develops in patients who had abdominal surgery. In rare instances, it occurs subsequent to gastrointestinal stromal tumor (GIST). This special situation has clinical significance in imatinib era. About 1000 patients with GIST in our institution from 1993 to 2010 were re-evaluated based on their clinical and pathological data, the treatment strategies and the follow-up information. We identified 2 patients who developed IAF after GIST resection. Patient 1 was a 54 year-old male and had 5 cm × 4.5 cm × 3.5 cm jejunal GIST excised on February 22, 1994. Three years later, an abdominal mass with 7 cm × 6 cm × 3 cm was identified. He was diagnosed as recurrent GIST from clinical point of view. After excision, the second tumor was confirmed to be IAF. Patient 2 was a 45-year-old male and had 6 cm × 4 cm × 3 cm duodenal GIST excised on August 19, 2008. One year later, a 4 cm mass was found at the original surgical site. The patient refused to take imatinib until the tumor increased to 8 cm six months later. The tumor continued to increase after 6 months’ imatinib therapy, decision of surgical resection was made by multidisciplinary team. The second tumor was confirmed to be IAF with size of 17 cm × 13 cm × 11 cm. Although IAF subsequent to GIST is very rare, it is of clinical significance in imatinib era as an influencing factor for making clinical decision.
Virtual slides
The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1076715989961803
doi:10.1186/1746-1596-8-125
PMCID: PMC3751073  PMID: 23902675
GIST; Intra-abdominal fibromatosis (IAF); Imatinib
20.  Safety and efficacy of a non-polymeric paclitaxel-eluting microporous stent in real-world percutaneous coronary intervention 
At present, there is an increasing focus on stents that have a biodegradable polymer coating, rather than a permanent polymer coating. This is due to the fact that following the implantation of a drug-eluting stent (DES) with a permanent polymer coating, the continued existence of the coating may result in a foreign body reaction and delayed re-endothelialization. The aim of the present study was to evaluate the safety and efficacy of a non-polymeric paclitaxel-eluting microporous (YINYI™) stent in real-life percutaneous coronary intervention (PCI) for patients with coronary artery disease (CAD). A total of 686 YINYI™ stents were implanted in 404 patients with CAD in a PCI procedure and outpatient follow-ups were performed 1, 6, 12 and 15 months subsequent to the PCI, respectively. The observation endpoints were major adverse cardiac events (MACEs), including cardiac death, non-fatal myocardial infarction (MI), restenosis, target lesion revascularization, stent thrombosis and recurrence of angina pectoris. The average follow-up time was 15 months. The results revealed that the cumulative incidences of MACEs were as follows: mortality, 0.99%; non-fatal MI, 0.74%; restenosis, 4.0%; and target lesion revascularization, 2.7%. The results at the short- and long-term clinical follow-ups indicated that YINYI™ stents are effective and safe for use in PCI for patients with CAD.
doi:10.3892/etm.2013.1217
PMCID: PMC3786829  PMID: 24137271
coronary artery disease; percutaneous coronary intervention; YINYI™ stent; non-polymeric paclitaxel-eluting microporous stent; follow-up
21.  Fatty liver index correlates with non-alcoholic fatty liver disease, but not with newly diagnosed coronary artery atherosclerotic disease in Chinese patients 
BMC Gastroenterology  2013;13:110.
Background
Fatty liver index (FLI) was recently established to predict non-alcoholic fatty liver disease (NAFLD) in general population, which is known to be associated with coronary artery atherosclerotic disease (CAD).
This study aims to investigate whether FLI correlates with NAFLD and with newly diagnosed CAD in a special Chinese population who underwent coronary angiography.
Methods
Patients with CAD (n = 231) and without CAD (n = 482) as confirmed by coronary angiography were included. Among them, 574 patients underwent B-ultrosonography were divided into NAFLD group (n = 209) and non-NAFLD group (n = 365). Correlation between FLI and NAFLD was analyzed using pearson’s correlation. The associations between FLI and NAFLD as well as CAD were assessed using logistic regression. The predictive accuracy of FLI for NAFLD was evaluated using receiver operating characteristics (ROC) curve analysis.
Results
FLI was significantly higher in NAFLD group (37.10 ± 1.95) than in non-NAFLD group (17.70 ± 1.04), P < 0.01. FLI correlated with NAFLD (r = 0.372, P < 0.001). The algorithm for FLI had a ROC-AUC of 0.721 (95% CI: 0.678–0.764) in the prediction of NAFLD. Logistic regression analysis showed that FLI was associated with NAFLD (adjusted OR = 1.038, 95% CI: 1.029-1.047, P < 0.01). The proportion of patients with CAD did not differ among the groups of FLI ≤ 30 (32.3%), 30-60 (31.0%), and ≥60 (35.3%). No significant association was found between FLI and CAD (adjusted OR = 0.992, 95% CI: 0.981-1.003 in men and OR = 0.987, 95% CI: 0.963-1.012 in women, P > 0.05).
Conclusions
FLI showed good correlation with NAFLD in patients who underwent coronary angiography, but not with newly diagnosed CAD. This might be underestimated because some patients in non-CAD group may have other underlying cardiovascular diseases.
doi:10.1186/1471-230X-13-110
PMCID: PMC3710104  PMID: 23834773
Fatty liver index; Non-alcoholic fatty liver disease; Coronary artery atherosclerotic disease; Coronary angiography
22.  Intraspinal AAV Injections Immediately Rostral to a Thoracic Spinal Cord Injury Site Efficiently Transduces Neurons in Spinal Cord and Brain 
In the vast majority of studies utilizing adeno-associated virus (AAV) in central nervous system applications, including those published with spinal cord injury (SCI) models, AAV has been administered at the level of the cell body of neurons targeted for genetic modification, resulting in transduction of neurons in the vicinity of the injection site. However, as SCI interrupts many axon tracts, it may be more beneficial to transduce a diverse pool of supraspinal neurons. We determined if descending axons severed by SCI are capable of retrogradely transporting AAV to remotely transduce a variety of brain regions. Different AAV serotypes encoding the reporter green fluorescent protein (GFP) were injected into gray and white matter immediately rostral to a spinal transection site. This resulted in the transduction of thousands of neurons within the spinal cord and in multiple regions within the brainstem that project to spinal cord. In addition, we established that different serotypes had disparate regional specificity and that AAV5 transduced the most brain and spinal cord neurons. This is the first demonstration that retrograde transport of AAV by axons severed by SCI is an effective means to transduce a collection of supraspinal neurons. Thus, we identify a novel, minimally invasive means to transduce a variety of neuronal populations within both the spinal cord and the brain following SCI. This paradigm to broadly distribute viral vectors has the potential to be an important component of a combinatorial strategy to promote functional axonal regeneration.
doi:10.1038/mtna.2013.34
PMCID: PMC3731889  PMID: 23881451
adeno-associated virus; propriospinal; retrograde transduction; spinal cord injury; supraspinal
23.  Risk of bladder cancer in patients with diabetes mellitus: an updated meta-analysis of 36 observational studies 
BMC Cancer  2013;13:310.
Background
Increasing evidence suggests that a history of diabetes mellitus (DM) may be associated with an increased risk of bladder cancer. We performed a systematic review with meta-analysis to explore this relationship.
Methods
We identified studies by a literature search of Medline (from 1 January 1966) and EMBASE (from 1 January 1974), through 29 February 2012, and by searching the reference lists of pertinent articles. Summary relative risks (RRs) with corresponding 95% confidence intervals (CIs) were calculated with a random-effects model.
Results
A total of 36 studies (9 case–control studies, 19 cohort studies and 8 cohort studies of patients with diabetes) fulfilled the inclusion criteria. Analysis of all studies showed that DM was associated with an increased risk of bladder cancer (the summary RR = 1.35, 95% CI 1.17–1.56, p < 0.001, I2 = 94.7%). In analysis stratified by study design, diabetes was positively associated with risk of bladder cancer in case–control studies (RR = 1.45, 95% CI 1.13-1.86, p = 0.005, I2 = 63.8%) and cohort studies (RR = 1.35, 95% CI 1.12-1.62, p < 0.001, I2 = 94.3%), but not in cohort studies of diabetic patients (RR = 1.25, 95% CI 0.86–1.81, p < 0.001, I2 = 97.4%). The RRs of bladder cancer were 1.38 (1.08-1.78) for men and 1.38 (0.90-2.10) for women with diabetes, respectively. Noteworthy, the relative risk of bladder cancer was negatively correlated with the duration of DM, with the higher risk of bladder cancer found among patients diagnosed within less than 5 years.
Conclusions
These findings support the hypothesis that men with diabetes have a modestly increased risk of bladder cancer, while women with diabetes were not the case.
doi:10.1186/1471-2407-13-310
PMCID: PMC3699355  PMID: 23803148
Diabetes; Bladder Cancer; Meta-analysis
24.  The GSTP1 105Val Allele Increases Breast Cancer Risk and Aggressiveness but Enhances Response to Cyclophosphamide Chemotherapy in North China 
PLoS ONE  2013;8(6):e67589.
The glutathione-S-transferase (GST) family contributes to the inactivation of various toxic compounds formed as secondary metabolites during oxidative stress. GSTP1 accounts for the majority of the GST family enzymatic activity, and the activity of GSTP1 enzyme can be altered by the presence of the Ile105Val polymorphism. In this study, we examined the polymorphic frequency of GSTP1 Ile105Val genotype in 920 breast cancer patients and 783 healthy controls in women of North China. Results showed that GSTP1 105Val allele (Ile/Val and Val/Val) was associated with a higher breast cancer risk (OR = 1.38, 95% CI: 1.14–1.69; P = 0.001) and more aggressive tumors with histological grade III (OR = 1.15, 95% CI: 1.05–1.26; P = 0.001), lymph node metastases (OR = 2.35, 95% CI: 1.72–3.21; P < 0.001), as well as ER negative (OR = 1.77, 95% CI: 1.31–2.39; P < 0.001) than those carrying the Ile/Ile allele. However, the patients with the GSTP1 105Val genotype had a better disease free survival after cyclophosphamide (CTX)-based chemotherapy than those with Ile/Ile (HR = 0.77, 95% CI: 0.45–0.91; P < 0.001). Furthermore, in vitro cellular experiments demonstrated that breast cancer cells with the GSTP1 105Val allele were significantly more sensitive to CTX-induced proliferation inhibition. Thus, we conclude that the GSTP1 105Val allele increases breast cancer risk and aggressiveness and enhance response to CTX-based chemotherapy in women of North China. Detection of the GSTP1 Ile105Val genotype may help screen for high-risk populations and direct individualized therapy.
doi:10.1371/journal.pone.0067589
PMCID: PMC3691318  PMID: 23826324
25.  The Composition of Microbiome in Larynx and the Throat Biodiversity between Laryngeal Squamous Cell Carcinoma Patients and Control Population 
PLoS ONE  2013;8(6):e66476.
The throat is an ecological assemblage involved human cells and microbiota, and the colonizing bacteria are important factors in balancing this environment. However, this bacterial community profile has thus been poorly investigated. The purpose of this study was to investigate the microbial biology of the larynx and to analyze the throat biodiversity in laryngeal carcinoma patients compared to a control population in a case-control study. Barcoded pyrosequencing analysis of the 16S rRNA gene was used. We collected tissue samples from 29 patients with laryngeal carcinoma and 31 control patients with vocal cord polyps. The findings of high-quality sequence datasets revealed 218 genera from 13 phyla in the laryngeal mucosa. The predominant communities of phyla in the larynx were Firmicutes (54%), Fusobacteria (17%), Bacteroidetes (15%), Proteobacteria (11%), and Actinobacteria (3%). The leading genera were Streptococcus (36%), Fusobacterium (15%), Prevotella (12%), Neisseria (6%), and Gemella (4%). The throat bacterial compositions were highly different between laryngeal carcinoma subjects and control population (p = 0.006). The abundance of the 26 genera was significantly different between the laryngeal cancer and control groups by metastats analysis (p<0.05). Fifteen genera may be associated with laryngeal carcinoma by partial least squares discriminant analysis (p<0.001). In summary, this study revealed the microbiota profiles in laryngeal mucosa from tissue specimens. The compositions of bacteria community in throat were different between laryngeal cancer patients and controls, and probably were related with this carcinoma. The disruption of this bio-ecological niche might be a risk factor for laryngeal carcinoma.
doi:10.1371/journal.pone.0066476
PMCID: PMC3688906  PMID: 23824228

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