Objective: The purpose of this study is to evaluate the potential diagnostic benefit of SPM-based semi-quantitative FDG-PET analysis in autoimmune encephalitis (AE) compared with visual analysis by experienced neuroradiologists using a larger sample size.Methods: This observational retrospective case series study was conducted from a tertiary epilepsy center between May 2014 and March 2017. Healthy individuals without any neurologic or psychiatric diseases were recruited as control. We determined brain FDG-PET abnormal glucose metabolism on medial temporal lobe and basal ganglia using semi-quantitative analysis and compared this method with visual analysis at the same time among patients with autoantibody positive AE.Results: Twenty-eight patients with clinically diagnosed AE and 53 healthy individuals without any neurologic or psychiatric diseases were recruited. On the medial temporal lobe and the basal ganglia, semi-quantitative analysis showed consistency with the visual assessment for whom they had abnormal metabolism by visual assessment. More importantly, 56% patients on medial temporal lobe and 73% patients on the basal ganglia respectively who were not identified by visual inspection can be detected by semi-quantitative analysis, demonstrating the greater sensitivity of semi-quantitative analysis compared with visual assessment.Significance: This study showed semi-quantitative brain FDG-PET analysis was better than visual analysis in view of observing the abnormal glucose metabolism of patients with autoantibody positive AE. Semi-quantitative FDG-PET analysis appears to be a helpful tool in early diagnosis of patients with AE. The clinical diagnosis of autoimmune encephalitis (AE) can be difficult since patients manifest various unspecific symptoms that overlap with other encephalitis such as infectious encephalitis . The cliRecently, leading authorities on the diagnosis and treatment of AE convened and recommended criteria for the diagnoses of possible, probable and definite AE to mitigate the delay in initiation of therapy while awaiting autoantibody assay results . FrancesTraditionally, whole body 18F-FDG-PET has been utilized to assess for occult malignancy as a cause for AE. Recent publications have begun to explore the added value of brain 18F-FDG PET/CT in evaluating these patients –13. SeveDespite visual analysis is often the first step of brain 18F-FDG PET reading, the standardization in view of reading images and reporting results is defective now . In clinIn the present study, we modified the semi-quantitative FDG-PET analysis as a biomarker for definite, autoantibody positive AE, allowing for additional region of interest (ROI) analyses of the mesial temporal lobe and basal ganglia. We used this analysis method in patients with antibody positive AE. The aim was to evaluate the potential diagnostic value of this analysis method compared with visual analysis by experienced neuroradiologists.t(79) = 1.97; p > 0.05). The detailed demographics of participants were presented in We recruited 28 AE patients with autoantibody  in our Tertiary Epilepsy Center from May 2014 to April 2017. All patients who had undergone cerebral FDG-PET were included in this retrospective study. Fifty-three healthy individuals  without any neurologic or psychiatric diseases were recruited. The age of these two group of participants were well matched and showed no significant difference ; (2) brain MRI or FDG-PET showing signal changes consistent with limbic encephalitis; (3) autoimmune/paraneoplastic antibodies in serum and/or CSF which have been associated with autoimmune encephalitis in previous studies . PET/CT images were acquired with the use of a multidetector helical PET/CT scanner . All cerebral FDG-PET studies were done in conjunction with whole-body PET scans .The brain Previous study demonstrated that AE were usually associated with the abnormalities of glucose metabolism in some brain regions, such as medial temporal lobe and basal ganglia . Thus, thttps://www.fil.ion.ucl.ac.uk/spm/software/spm12/). During the preprocessing, the co-registration between PET metabolism images and CT structural images was firstly performed. Then, the CT images with high resolution were spatial normalized into Montreal Neurological Institute template (MNI). Thus, the PET images were also normalized into this standard templates using the computed space transformation for CT normalization. Finally, the PET scans were resampled to 2 × 2 × 2 mm3 voxels and spatially smoothed using an isotropic 6 mm full-width-half-maximal (FWHM). Additionally, to remove the bias of global metabolism, each voxel's intensity of PET scans was normalized by dividing the average of the voxels within the highest 20% of intensity . Whereas, the medial temporal lobe were identified hypo-metabolism if any of ROIs within the medial temporal lobe showed a significant lower metabolism than healthy group . For each ROI within basal ganglia, the same statistical analysis as the ROIs within medial temporal lobe was conducted on each patient.For each ROI within medial temporal lobe, the z-score was calculated as an indicator of abnormalities of glucose metabolism of each patient deviated from that of the control healthy group. Specifically, we firstly calculated the mean of whole voxels' intensities within each ROI to obtain the average metabolic of the ROI for each participant. Then, for each ROI, we calculated the mean (M) and standard deviation (SD) of the average metabolic cross all healthy individuals. Finally, for each patient, z-score was obtained by subtracting M from each ROI and then dividing the SD . The medial temporal lobe were identified hyper-metabolism if any of ROIs within the medial temporal lobe showed a significant higher metabolism than healthy group . Five patients among the 17 patients successfully detected by visual assessment showed a comparable hyper-metabolism between the right hemisphere and the left hemisphere. However, four patients showed worse abnormalities of glucose metabolism in the right hemisphere than the left hemisphere, while the other seven patients showed worse abnormalities of glucose metabolism in the left hemisphere than the right hemisphere. Besides, one patient among the 17 patients showed the hyper-metabolism only in the left hemisphere.The results of semi-quantitative analysis were also shown in On basal ganglia, 25 patients were identified as abnormal glucose metabolism. However, the other three patients weren't identified as abnormality. Thus, the sensitivity of semi-quantitative analysis on basal ganglia was about 89% (25/28).On the medial temporal lobe, as shown in More importantly, five of nine patients who were not identified by visual inspection (56%) were detected by semi-quantitative analysis, demonstrating the greater sensitivity of semi-quantitative analysis compared with visual assessment. Three of these five patients were identified as a bilateral abnormality and one patient was found to be abnormal only on left medial temporal lobe, and the other patient was found hyper-metabolism on the right medial temporal lobe while hypo-metabolism on the left medial temporal lobe.On the basal ganglia, as shown in More importantly, eight of 11 patients not identified by visual inspection (73%) were detected by semi-quantitative analysis, demonstrating the greater sensitivity of semi-quantitative analysis compared with visual assessment. Four of these eight patients were identified as bilateral hyper-metabolism, and Four of these eight patients were identified as bilateral hyper-metabolism, and one patient (subject 18) was found hyper-metabolism only on the left basal ganglia, and one patient (subject 16) was found hyper-metabolism only on the right basal ganglia, and one patient (subject 17) was found hyper-metabolism on the right basal ganglia while hypo-metabolism on the left basal ganglia, and the other one patient (subject 5) was found hypo-metabolism on the bilateral basal ganglia.t-test. On the medial temporal lobe, the mean duration of detected group was 2.33 ± 1.71 months, whereas the mean duration of missed group was 7.75 ± 1.71 months. On the basal ganglia, the mean duration of detected group was 2.60 ± 2.10 months, whereas the mean duration of missed group was 7.33 ± 2.31 months. These results suggested that the patients without abnormal metabolism through semi-quantitative analysis had significantly longer disease duration  by semi-quantitative analysis; in the other group of patients abnormal metabolism cannot be detected (missed group) by semi-quantitative analysis. We analyzed the disease duration of the two groups of patients using the independent  Seen in .To date, FDG-PET/CT is one of largest increases in the numbers of medical imaging scanners . MoreoveSince early immunotherapy can lead to better prognosis in AE, recent criteria have been improved to get early diagnosis . FDG-PETIn the present study, we described semi-quantitative brain FDG-PET findings among patients with autoantibody positive AE, and compared this method with visual analysis at the same time. Our results showed that semi-quantitative brain FDG-PET analysis can find abnormal metabolism more sensitively, suggesting that semi-quantitative brain FDG-PET analysis was better than visual analysis. This study included a larger proportion of patients with LGI1. Previous case studies found striatal hyper-metabolism and/or the medial temporal lobe hyper-metabolism in patients with LGI1 encephalitis , 31. AccBR and anti-amphiphysin encephalitis as well as anti-LGI1 encephalitis all belong to limbic encephalitis. In addition, our previous study also showed that anti-GABABR encephalitis and anti-amphiphysin encephalitis had similar clinical manifestation to anti-LGI1 encephalitis  encephalitis has been described previously , 35, 36 Although semi-quantitative analysis can detect abnormal metabolism more sensitively compared with visual analysis, the positive rate can only reach 50% on medial temporal lobe and 67% on basal ganglia, respectively. We suppose that is due to the different disease duration. Previous studies showed that FDG-PET abnormal metabolism can change with disease evolution. Intense 18F-FDG uptake can be found in bilateral limbic system at active disease status, and then the 18F-FDG uptake decreased gradually and eventually returned to normal following the clinical improvement after treatment , 13. In Limitations of this study include the retrospective design, the significant different disease duration among the patients and relative small sample size of normal metabolism by visual analysis. Because abnormal metabolism was associated with different disease status, this will lead to misunderstandings of abnormal status. Although the whole sample size of AE was large, the patients with normal metabolism by visual analysis were relatively few, this need be evaluated by enrolling more patients in the future. Moreover, the patients were enrolled at a specialized tertiary epilepsy center, making the study subject to referral bias.This study which showed semi-quantitative brain FDG-PET analysis was better than visual analysis in view of observing the abnormal glucose metabolism of patients with antibody positive AE. Semi-quantitative FDG-PET analysis appears to be a helpful tool in early diagnosis of patients with AE, especially those with normal MRI scans. Further research is needed to validate the sensitivity and specificity of semi-quantitative FDG-PET analysis in the early diagnosis of patients with AE. Besides, its usefulness for a better characterization of specific syndromes and their clinical course and response to therapy also needs to be further evaluated in prospective studies.Informed consent to participate the study and for publication for clinical details were obtained from each subject enrolled. The study was approved by the Medical Ethics Committee of Beijing Tiantan Hospital, Capital Medical University and was carried out in accordance with the Declaration of Helsinki.R-JL and JL contributed to the study concept and design, and critically revised the manuscript for important intellectual content. R-JL, X-QS, QW, and X-BZ acquired the data. JP, GZ, JL, and R-JL analyzed and interpreted the data. R-JL, JP, and JL drafted the manuscript and provided statistical expertise. QW and X-QS provided administrative, technical, and material support. X-BZ and JL supervised the study.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Prostate cancer (PCa) is one of the most commonly diagnosed cancers worldwide, accounting for almost 1 in 5 new cancer diagnoses in the US alone. The current non-invasive biomarker prostate specific antigen (PSA) has lately been presented with many limitations, such as low specificity and often associated with over-diagnosis. The dysregulation of miRNAs in cancer has been widely reported and it has often been shown to be specific, sensitive and stable, suggesting miRNAs could be a potential specific biomarker for the disease. Previously, we identified four miRNAs that are significantly upregulated in plasma from PCa patients when compared to healthy controls: miR-98-5p, miR-152-3p, miR-326 and miR-4289. This panel showed high specificity and sensitivity in detecting PCa (area under the curve (AUC) = 0.88). To investigate the specificity of these miRNAs as biomarkers for PCa, we undertook an in depth analysis on these miRNAs in cancer from the existing literature and data. Additionally, we explored their prognostic value found in the literature when available. Most studies showed these miRNAs are downregulated in cancer and this is often associated with cancer progression and poorer overall survival rate. These results suggest our four miRNA signatures could potentially become a specific PCa diagnostic tool of which prognostic potential should also be explored. Prostate cancer (PCa) is the second most commonly diagnosed cancer worldwide with more than 1.1 million new cancer cases each year, accounting for 15% of all cancers diagnosed . AlthougMicroRNAs (miRNAs) are a class of small, non-coding RNA molecules. They bind to their target mRNAs to regulate gene expression  and someWe have recently identified a panel of four miRNAs: miR-98-5p, miR-152-3p, miR-326 and miR-4289 ,28. To fMore than 300 abstracts were retrieved initially from PubMed. These included information for the four miRNAs of interest related to their dysregulation in cancer and/or survival. Those studies with less than 20 patients were discarded, leaving ~200 abstracts for further inspection. Papers with no clinical and pathological data and those that focused on the basic biology of the miRNA were excluded. This left 78 studies to be examined in further detail to extract potential data. Of these, 38 research articles reported significant results for the miRNAs of interest for their association with one or more forms of cancer, from which data was subsequently extracted and presented in this article.The main focus of this section was to compile recent reported dysregulation expression data for our miRNA signature in all cancers and assess its specificity for PCa. Cancer-related dysregulation that met our selection criteria was found for three of the miRNAs, with no reported data for miR-4289. However, two examples have been included at the end of this section to get a preliminary appreciation of what is available for miR-4289 . Additio−8) . Th. Th26]. The research done on miR-4289 is currently very limited, with only a few studies reporting its role in cancer. Because of this availability shortage, two studies are here mentioned despite of not fitting the search criteria applied. Dong et al. undertook miRNA profiling in three samples only to detect differentially expressed miRNAs between GBM patients and healthy controls. From the 752 miRNAs analysed, 115 miRNAs were found to be upregulated being miR-4289 one of them (P = 0.0002) . The secNo reported survival data in cancer was found for the miR-4289.N = 48), miR-152-3p (N = 49) and miR-326 (N = 49) and are presented in Differential expression data reported in dbDEMC 2.0 was found for three out of the four miRNAs  and all Additional information such as the experiment ID, Gene Expression Omnibus (GEO)/TCGA information, cancer subtypes, T values, P values and β values and adjusted P values are all summarised in The literature research previously presented also provided a number of validated targets for the different miRNAs that compose the signature. The method of validation, gene target effect and involved pathway or molecular function are summarised in A good usable biomarker has to be selective and objectively measured to evaluate diverse biological or pathogenic processes, as described by the Biomarker Consortium group . AmongstOur own analysis showed significantly higher expression plasma levels of miR-98-5p, miR-152-3p, miR-326 and miR-4289 in prostate cancer patients when compare to healthy individuals in two cohorts and this was associated with an increased risk of developing the disease for the first three miRNAs. Opposite results were obtained for miR-4289 between the two cohorts analysed. From our literature search, the upregulation of miR-98 and miR-326 in PCa suggests them to be PCa-specific when compared to other cancers. By contrast, miR-152 lacked specificity for this disease and not enough data for miR-4289 was available to draw any conclusions. Most studies are on tumor samples in PCa. Interestingly, the upregulation of miR-98 in recurrent PCa patients after radical prostatectomy when compared to non-recurrent patients was also reported , placingN = 100) when compared to healthy tissue (N = 14) with a highly hyper-methylated promoter region [A potential limitation of this study is the lack of a standardised miRNA extraction protocol from clinical samples available in the scientific community. As shown recently ,79, deter region , making r region ,68 and ir region ,90.In summary, cancer expression data has been reported for miR-98, miR-152 and miR-326. From the research articles included, the upregulation of miR-98 and miR-326 in PCa was shown to be highly specific. In most cases, these three miRNAs were downregulated in cancer and this was often associated with a poorer prognosis, later staging, increased tumor size, and higher pathological grades. We have reported an upregulation of the miRNA signature miR-98-5p, miR-152-3p, miR-326 and miR-4289 in PCa plasma patient samples when compared to healthy controls. Since our signature’s upregulation has been detected in plasma, and most of the dysregulated expression data comes from tissue samples, the results presented here should be taken as a summary of what data is available for these four miRNAs expressions in cancer to date. Indeed, further studies are needed to clarify their plasma and serum dysregulations in different cancers and their suitability as specific markers of PCa.The literature used within this study was obtained via the PubMed database, utilising studies conducted in the last 10 years and the strategy followed is summarised in http://www.picb.ac.cn/dbDEMC) (data extracted in March 2018) [In addition to the reported data in the literature for this signature, we extracted normalised logarithmic fold change miRNA expression data from the dedicated Differentially Expressed miRNAs in human Cancers database  . This inThose studies that reported survival data and/or expression in metastatic tissue when compared to non-metastatic disease are summarised in The upregulation of miR-98 and miR-326 in PCa were shown to be highly specific of prostate cancer as in the majority of the cases reported these two miRNAs were downregulated in cancer and they were often associated with a poorer prognosis that included later staging, larger tumor size, and higher pathological grades. The miRNA signature we have recently reported showed that miR-98-5p, miR-152-3p, miR-326 and miR-4289 were upregulated in PCa plasma patient samples when compared to healthy controls. A limitation of our study includes the nature of our samples (plasma) which differs from most of the samples’ origin found in the literature (tissue). Therefore, here we present a snapshot of the significant reported dysregulation in cancer of these four miRNAs to date.


Dyslipidemia is a major risk factor for the development of cardiovascular disease. Both statins and omega-3 fatty acids demonstrate beneficial effects on lipid concentrations. The goal was to evaluate the safety and efficacy of combination therapy with statins and omega-3 fatty acids.We performed a systematic review and meta-analysis of published data to compare the safety and efficacy of combination therapy with statins and omega-3 fatty acids versus statin monotherapy in patients with dyslipidemia. Six articles were assessed in the present meta-analysis (quantitative assessment) and qualitative assessment.In terms of efficacy, the combination treatment afforded a significantly greater reduction in total cholesterol/high-density lipoprotein cholesterol than statin alone did . However, there was no significant difference in low-density lipoprotein (LDL) cholesterol between the 2 groups. Qualitative assessment of other lipid parameters was performed. Combination therapy with statins and omega-3 fatty acids was generally more effective on lipid concentration than statin monotherapy. In terms of safety, there were no significant differences in total adverse events between the 2 groups. Gastrointestinal adverse events were found to be significantly increased in patients receiving combination therapy using the fixed-effects model .We suggest that combination therapy with statins and omega-3 fatty acids enhances lipid profile, except LDL cholesterol, compared with statin monotherapy. Nevertheless, statin and omega-3 fatty acid combination should be cautiously recommended, taking into account the clinical importance of LDL cholesterol and safety issues associated with their concomitant use. It is one of the major risk factors for the development of coronary heart disease. The condition should therefore be controlled to prevent primary or recurrent cardiovascular events. Several pharmacotherapeutic options are available to control lipid concentrations and reduce cardiovascular death and total mortality. Combination therapy may be considered in patients not responding to statin monotherapy. In such cases, fibrates, niacin, bile acid resins, or ezetimibe are used concomitantly or alone; however, the patients should be monitored closely for the development of adverse effects and drug interactions.Statins are the initial treatment choice for dyslipidemia in most patients. They are hydroxymethylglutaryl coenzyme A reductase inhibitors and have potent total and LDL cholesterol-lowering effects.–4 Lipid profile changes, including reduction in cholesterol and triglycerides, by omega-3 fatty acids have been reported in normal volunteers and patients with hyperglyceridemia.,6 Therefore, omega-3 fatty acids may be useful in patients with elevated triglyceride levels; however, the clinical role of omega-3 fatty acids is not well defined. Moreover, the safety and efficacy of combination therapy with omega-3 fatty acids and statins have not been evaluated adequately.Omega-3 polyunsaturated fatty acids are also used as an alternative therapy to regulate abnormal lipid concentrations. The efficacy of fish oil supplementation containing high omega-3 fatty acids on lipid metabolism is mediated through the reduction in very low-density lipoprotein (VLDL) production and suppression of VLDL and apolipoprotein B (apoB).–12 Nevertheless, the results of trials comparing the efficacy of statin monotherapy versus combination therapy with omega-3 fatty acids have not yet been integrated into clinical decision making. In addition, the safety of combination therapies remains controversial.Recently, several clinical trials have shown that combination therapy with statins and omega-3 fatty acids improved lipid profiles and prevented cardiovascular events.Therefore, we performed a systematic review and meta-analysis to determine the changes in lipid concentrations and the number of adverse events in the reported studies by comparing the effects of statin monotherapy versus combination therapy with statins and omega-3 fatty acids.22.1hydroxymethylglutaryl-CoA reductase inhibitors, statins, omega-3 polyunsaturated fatty acids, omega-3, omega-3 fatty acids, dyslipidemia, hypercholesterolemia, and hyperglyceridemia. The bibliographies of retrieved articles and relevant reviews were searched to identify additional eligible studies. We did not impose any publication or language limitations. The search was completed on January 15, 2017.We searched for published articles comparing the lipid-lowering effects and safety of statins and omega-3 fatty acids in patients with dyslipidemia. We searched online databases, including MEDLINE (OVID and PubMed) and the Cochrane Library. The search terms were combinations of the following PubMed MeSH terms and related text terms: 2.2Two authors independently reviewed and selected studies for inclusion in the systematic review. The inclusion criteria were as follows: the study was a randomized clinical trial; compared the efficacy and safety of fibrates and statins; included measurements of lipid profiles; and described a number of adverse events. Any disagreement in terms of inclusion of an article for evaluation was resolved by discussion. If a trial was described in more than one report, we extracted data from the most complete account and used the other publications only to clarify those data.2.3Detailed reviews of full-text articles were performed independently by 2 authors. The following data were extracted from each study: the first author's surname; the year of publication; the country in which the work was performed; the number of participants; patient characteristics ; the treatment given (regimen and period); and changes in serum lipid concentrations.2.4 The Jadad scale evaluates randomized controlled trials using the following 5 indicators: an adequate description of how randomization was achieved; the appropriateness of the randomization method; an adequate account of how the investigators were double blinded; the appropriateness of the double-blinding method chosen; and details on patient withdrawal and dropout. A score >3 was considered to reflect high-quality work. Any disagreement between the 2 authors was resolved by discussion.The methodological quality of each trial was evaluated by 2 authors with the aid of the Jadad scale.2.52.5.1In terms of evaluation of efficacy, the endpoints of analysis were changes in lipid concentrations, including changes in total cholesterol, LDL cholesterol, HDL cholesterol, triglycerides, and other related lipid concentrations. For the meta-analysis, the change in LDL cholesterol and total cholesterol/HDL cholesterol ratio, respectively, was calculated as the difference between the baseline and final measurements performed in each study group. Each mean change, with a 95% confidence interval (CI), was calculated to allow assessment of the lipid-lowering effects of statin monotherapy or combination therapy with omega-3 fatty acids.2.5.2To evaluate treatment safety, we counted the total number of adverse events and gastrointestinal adverse events in each study group, and compared these values between treatments. Relative risk (RR) values and 95% CIs were calculated to compare the frequencies of adverse events associated with the use of statin monotherapy or combination therapy with omega-3 fatty acids.2.62 test (employing Q statistics) and quantified by calculating I2 values. A fixed-effects model (the Mantel-Haenszel method) was used in the analysis. The results were compared to those yielded by a random-effects model (the Der Simonian-Laird method).Study heterogeneity was assessed using the χ,18Sensitivity analyses were performed by excluding the contribution of each study to the meta-analysis data in turn. The potential existence of publication bias was examined using Begg and Egger tests.P < .05 was considered to indicate statistical significance.All statistical analyses were performed using the Comprehensive Meta-analysis Software version 2 . All statistical tests were 2 sided and a value of 2.7This systematic review was performed based the previously reported articles. Therefore, any ethical review or approval is not applicable.33.1 Using the Jadad system, 2 studies were classified as low quality (a score of ≤2) and 4 studies as high quality (scores of ≥3).One hundred eighty-six articles were identified by the literature search. After removal of duplicates, the titles and abstracts of 137 articles were screened. Of these, 100 articles were excluded, and the full texts of the remaining 37 articles were assessed in terms of eligibility. A further 29 articles were excluded, and data from the remaining 6 articles were included in the present systematic review  treated with statin alone, and 265 adverse events occurred in 611 patients 43.4%) treated with statin plus omega-3 fatty acid  treated with statin alone and 95 occurred in 611 patients (15.5%) treated with statin plus omega-3 fatty acid . We also evaluated publication bias. The funnel plots were not obviously asymmetric. The results of Begg rank-correlation test and Egger regression test are shown in Table 44.1We performed a systematic review and meta-analysis to evaluate the safety and efficacy of statin monotherapy and combination therapy with statin and omega-3 fatty acid in patients with dyslipidemia. First, we found that changes in lipid concentration differed significantly between the 2 drug regimens. Combination therapy with statin and omega-3 fatty acid afforded greater reduction in total cholesterol/HDL cholesterol ratio. Second, treatment with statin plus omega-3 fatty acid was associated with a significant increase in the number of gastrointestinal adverse events, but not in total adverse events.–10,12 In all studies, the combination treatment afforded significantly greater reductions in non-HDL cholesterol and triglycerides than treatment with statin alone did. In addition, the combination treatment was associated with significant increases in HDL cholesterol. Five studies evaluating VLDL cholesterol showed significant cholesterol-lowering effects by the combination treatment.–11 Moreover, the combination treatment was found to be more effective in reducing total cholesterol than statin monotherapy in the 4 included studies.–10 Six studies were included to evaluate apoB, and 2 of the studies showed no significant difference between the 2 groups.–12 A significant association between combination treatment and apolipoprotein A1 increase was shown in 1 of the 4 included studies. Overall, the results of qualitative assessment showed that combination therapy with statin and omega-3 fatty acid was generally more effective in regulating lipid concentration than statin monotherapy.We performed the qualitative assessment of other lipid concentrations owing to lack of adequate number of studies or different data types to perform a meta-analysis Table . Five st,21 Based on the present meta-analysis, no significant difference in LDL cholesterol levels between statin monotherapy and combination therapy with omega-3 fatty acid was inferred; however, the offsetting effect of omega-3 fatty acid on LDL cholesterol cannot be completely excluded. This finding is in agreement with the results of previous studies in which omega-3 fatty acid consumption was reported to be associated with mild increases in LDL cholesterol in patients with hypertriglycemia and dyslipidemia.,6 This change can be explained, at least in part, by the increased rate of conversion of VLDL to LDL particles.–4LDL cholesterol is clinically the most important measurement in a lipid profile, and it is closely related to cardiovascular disease or death. are noteworthy. The meta-analysis was performed to estimate the total and cardiovascular mortalities in participants receiving omega-3 fatty acid, followed by a metaregression of dose, docosahexaenoic acid/eicosapentaenoic acid (DHA/EPA) ratio, duration of treatment, and use of lipid-lowering/statin therapy in control group. The results showed that omega-3 fatty acid had no effect on total mortality , but it reduced cardiovascular mortality . In addition, lower control group statin use and higher DHA/EPA ratio was associated with higher reduction in total mortality. In other words, statin use may mitigate the positive effects of omega-3 fatty acid. Further research or clinic trials can provide more evidence on the influence of combination therapy with statin and omega-3 fatty acid on LDL cholesterol.The results of the meta-analysis by Sethi et alIn the meta-analysis on safety, the incidence of gastrointestinal adverse events was slightly higher in patients receiving statin and omega-3 fatty acid combination therapy. Statin or omega-3 use is associated with several side effects ranging from mild to severe. Gastrointestinal problems are frequently caused by treatment with statins and omega-3 fatty acid alone, and their concomitant use may increase the risk of side effects significantly. Therefore, the adverse events should be monitored more intensively in patients receiving combination treatment.4.2There are certain limitations to our present meta-analysis. First, only a small number of relevant studies were included and data were inadequate. Second, our analysis was based on previous reports, which were not necessarily complete or accurate, and we were unable to analyze data, including sex, ages, duration, or other factors. Third, the results based on statistical analysis could be partially different from the evaluations of safety or efficacy in individual patients.Despite these limitations, to the best of our knowledge, this is the first study to compare the safety and efficacy of statin treatment alone versus combination therapy with statin and omega-3 fatty acid in patients with dyslipidemia. We derived accurate estimates of the clinical efficacies of the 2 types of treatment. Our data have greater statistical power than the data subsets of the reports included in the meta-analysis.5Overall, we suggest that combination therapy with statins and omega-3 fatty acid enhances the lipid profile, except LDL cholesterol, when compared with statin monotherapy. However, controlling LDL cholesterol levels is important for preventing cardiovascular diseases and related deaths. Furthermore, safety issues with the concomitant use of statins and omega-3 fatty acid should be considered. Thus, combination therapy with statins and omega-3 fatty acid should be cautiously recommended after assessing the benefits and risks.The contributions of the authors to the manuscript are as follows. Choi HD: study design, data collection, data analyses, coordination, and writing the manuscript; S.M.C.: study design, data collection, data analyses, and reviewing the manuscript. All authors read and approved the final manuscript.Conceptualization: Hye Duck Choi.Data curation: Hye Duck Choi, Seung Min Chae.Formal analysis: Hye Duck Choi, Seung Min Chae.Funding acquisition: Hye Duck Choi.Investigation: Hye Duck Choi, Seung Min Chae.Methodology: Hye Duck Choi.Project administration: Hye Duck Choi.Resources: Hye Duck Choi.Software: Hye Duck Choi.Supervision: Hye Duck Choi.Validation: Hye Duck Choi.Visualization: Hye Duck Choi.Writing – original draft: Hye Duck Choi.Writing – review and editing: Hye Duck Choi.Hye Duck Choi orcid: 0000-0003-2292-6827.


Diagnostic and Statistical Manual of Mental Disorders (Fifth Edition) criteria for methamphetamine dependence, with a mean (SD) body mass index of 24.7 (3.5) kg/m2 and age of 30.2 (5.1) years, were randomly assigned to one of two exercise groups: moderate intensity  and high intensity . Each group also performed a resting control session for 35 min 1 week before or after the exercise, in a counterbalanced order. Mean oxygenated hemoglobin concentration changes in the PFC when viewing visual food cues were assessed by fNIRS, and subjective feelings of appetite were self-rated using visual analog scales after moderate- or high-intensity aerobic exercise and after the resting control session. A continuous-wave NIRS device was used to obtain functional data: eight sources and seven detectors were placed on the scalp covering the PFC, resulting in 20 channels per participant. We found that moderate-intensity aerobic exercise significantly increased both, the activation of the left orbitofrontal cortex (OFC) to images of high-calorie food (P = 0.02) and subjective sensations of hunger  = 7.16, P = 0.01). To our knowledge, this study provides the first evidence that moderate-intensity aerobic exercise increases OFC activity associated with high-calorie food images and stimulates appetite in methamphetamine-dependent individuals. These changes suggest that exercise may reestablish the food reward pathway hijacked by drugs and restore sensitivity to natural rewards. This evidence may contribute to the development of specific exercise programs for populations with methamphetamine dependence.The brain prefrontal control system is critical to successful recovery from substance use disorders, and the prefrontal cortex (PFC) regulates striatal reward-related processes. Substance-dependent individuals exhibit an increased response to drug rewards and decreased response to natural, nondrug rewards. Short-term aerobic exercise can ameliorate craving and inhibitory deficits in methamphetamine users, but the effect of exercise on food reward is unknown. This study used functional near-infrared spectroscopy (fNIRS) to measure the effects of moderate- and high-intensity short-term aerobic exercise on prefrontal activity related to food images and recorded the subjective feelings of appetite in methamphetamine-dependent users. In total, 56 men who met the  Drug addiction is considered a chronic brain disease  regulates striatal reward-related processes and exhibits activity that predicts treatment outcome with respect to maintaining abstinence . Written informed consent was obtained from all participants before enrolling them in the study.2] who met the Diagnostic and Statistical Manual of Mental Disorders (Fifth Edition) criteria for MA dependence were recruited from the Drug Rehabilitation Bureau of Shi Liping in Zhejiang province. Participants were included if they met the following criteria: (1) no metabolic or chronic disease, no medical conditions, and not taking medication known to influence gastric emptying or appetite; (2) no eating disorder, psychiatric diagnoses, or neurological illness; (3) weight stable (<3 kg change in the last 3 months); (4) aged 18–45 years; (5) right-hand dominant; and (6) abstinent and receiving treatment for 3 months.In total, 56 men [mean and standard deviation (SD) age, 30.2 (5.1) years; mean (SD) body mass index (BMI), 24.7 (3.5) kg/mn = 28) and high-intensity (n = 28) exercise groups in demographic characteristics , fitness (BMI and resting heart rate), or drug use  prior to exercise intervention . We acquire dual-wavelength (760 and 850 nm) near-infrared light to measure the relative concentration changes in oxyHb and deoxyHb  using Matlab . Motion artifacts were detected within 0.5 s when the signal changed by more than 10% of the SD. Wavelet filtering was used in Homer2 to detect and remove these artifacts  at 50 rpm. Participants were allowed to warm-up for 5 min and to cool down for 5 min. The experimental exercise period was 25 min. For this period, the participant’s heart rate was maintained either within 65%–75% or within 76%–85% of their estimated maximum heart rate . Two-way repeated-measures analysis of variance was used to determine the main effects of exercise intervention (control and exercise) and exercise intensity (moderate and high) and their interaction effects on the PFC region activity and appetite measures. post hoc tests and controlling for multiple comparisons using the Benjamini-Hochberg false discovery rate procedure  = 6.66, P = 0.01) and of intensity  = 4.34, P = 0.04), and a significant interaction  = 5.33, P = 0.03) between exercise and intensity for Ch11. Furthermore, the HbO concentration changes increased significantly after moderate-intensity exercise (P = 0.02) but not high-intensity exercise  = 4.34, P = 0.04) and a significant interaction between exercise and intensity for Ch11  = 5.19, P = 0.03). The HbO concentration changes also increased significantly after moderate-intensity exercise (P = 0.02).Repeated measures analysis of variance results revealed significant main effects or interactions on the HbO concentration in 3 of 20 channels while participants were viewing high-calorie or low-calorie food images . After aexercise . We alsoSubjective feelings of appetite, in terms of hunger, fullness, and desire to eat after exercise and control sessions, are given in This study used fNIRS to examine the short-term effects of moderate- and high-intensity aerobic exercise on prefrontal brain activity and visual analog scale scores to assess the effects on appetite in individuals with MA dependence. The findings showed that activation of the OFC associated with viewing images of high-calorie foods increased in these individuals following moderate-intensity aerobic exercise compared with that at rest and that the subjective feeling of hunger increased in these individuals after exercise, especially after high-intensity exercise.Thus, consistent with our hypothesis, the present study found that short-term aerobic exercise increased OFC activation to high-calorie food cues in users of MA. Studies have found that individuals who are not drug users and are either normal weight or have obesity show different neural responses to visual food cues in the reward region; people with obesity have greater neural responses to food cues than people with normal weight, no matter whether they are hungry or full (Dimitropoulos et al., The present study showed that activation of the OFC associated with viewing images of high-calorie vs. low-calorie foods increased in these individuals following moderate-intensity aerobic exercise. This result is consistent with the finding that brain responses were positively associated with the self-rated desire to consume high-calorie food, in particular, savory food rather than sweet food. Our results also indicated that physical exercise was associated with the responsiveness of specific brain regions, and these brain region responses have been associated with the preference and desire for high-calorie foods (Killgore et al., The present study using neuroimaging is an early study of food reward among patients who are MA-dependent. By contrast, other such studies of human addiction have generally used healthy participants and focused on their neural responses to drug-related cues. Such studies have shown that drug-related cues activate the brain regions that are normally stimulated by nondrug-related rewards (Sell et al., Previous research suggests that the prefrontal control system may be the key to successful drug withdrawal. The frontal lobe, which has been shown to modulate processes associated with reward in the striatum, is one of the regions that predict treatment outcomes and has shown increased reward and cognitive control in patients who successfully remain abstinent (Garavan and Weierstall, Effects of exercise have been investigated in substance use disorder. Research has found that 10 min of exercise significantly reduces responses to images related to smoking in the OFC and dorsolateral PFC, as well as the subjective perception of cravings, consistent with reduced activation in food reward areas and better inhibitory control (Janse Van Rensburg et al., There are a number of limitations to consider in the evaluation of this study. First, our experiment was conducted in a drug rehabilitation center, with many practical and ethical restrictions regarding patient participation in studies. Therefore, it was not possible to bring people out of the center to conduct fMRI scans to measure neural activity in other brain regions. Second, The effect of an exercise intervention on food reward may vary according to an individual’s BMI (Rothemund et al., The present study reports a novel finding, to our knowledge, that short-term moderate-intensity aerobic exercise may increase neuronal responses related to food reward in the PFC region among persons who are MA-dependent. Exercise increased the appetite response, especially following high-intensity exercise. This suggests that moderate exercise may reestablish the food reward pathway acutely hijacked by drugs, restore sensitivity to natural rewards, and further promote drug withdrawal among drug-dependent users. This evidence may contribute to the development of specific exercise programs for MA-dependent populations.The datasets generated for this study are available on request to the corresponding author.The studies involving human participants were reviewed and approved by the ethics committee of the Shanghai University of Sport. The patients/participants provided their written informed consent to participate in this study.YZ and CZ contributed to the conception and design of the study. JW and YC conducted experiments and analyzed data. YZ wrote the first draft of the manuscript. HW and XL contributed to manuscript revision. All authors read and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Heroin addiction remains a significant public health problem worldwide, and relapse to heroin use following cessation of agonist maintenance treatment is common. The problems associated with use of opioid agonists mean that non-opioid therapies need to be developed to ameliorate acute and protracted opioid withdrawal syndromes.t test and Chi-square test. Data are compared between the 2 groups using repeated-measures analysis of variance, generalized estimating equations, and the paired t test.Fifteen men with opioid use disorder on methadone maintenance treatment have been enrolled from an addiction treatment center as an experimental group in this case-controlled study. This group is receiving laser meridian massage on the back, including the Bladder meridian and Governor Vessel, 3 times weekly for 4 weeks. An age-matched control group that does not receive laser meridian massage has also been enrolled. Urinary morphine levels are being checked before and after 2 and 4 weeks of treatment. Subjects are requested to self-report their number of episodes or days of heroin use and 0 to 10-point visual analogue scale scores for heroin craving/refusal to use heroin during the previous week before and after 2 and 4 weeks of treatment. Quality of life will be reported using the Short Form-12v2 before and after 4 weeks of treatment. Pulse diagnosis will be recorded and heart rate variability calculated after one single treatment session. The baseline patient characteristics will be compared between the experimental and control groups using the independent To investigate the effect of adjuvant laser meridian massage in men with opioid use disorder on methadone maintenance treatment.ClinicalTrials.gov NCT04003077. Methadone maintenance has been shown to reduce the frequency of opioid use, reduce mortality, reduce the transmission of human immunodeficiency virus and viral hepatitis, improve employment prospects, and reduce the frequency of criminal behavior. Many of these positive effects are strongly associated with higher daily methadone dose and with increasing duration of treatment. However, relapse to heroin use following cessation of agonist maintenance treatment is common. Research is lacking on when, who, and how to withdraw from opioid agonist maintenance treatment. The problems associated with use of opioid agonists have made it necessary to develop non-opioid therapies to ameliorate the symptoms of acute and protracted opioid withdrawal.,4Opioid use disorder (OUD), which most commonly manifests as heroin addiction, remains a significant public health problem worldwide. Clinically, OUD is characterized by physical dependence, as evidenced by tolerance and withdrawal, and by psychological symptoms, including drug cravings, depression, anxiety, and inability to control use of heroin.,6 Acupuncture is based on traditional Chinese medicine (TCM) theory and was developed according to the principle that human bodily functions are controlled by the “meridian” and “Qi and blood” systems. There are 365 designated acupuncture points located along 14 meridians that can be used to stimulate, balance, and harmonize the yin and yang by relieving blockages in the flow of Qi. This method of healing has been used to promote homeostasis of the body's organs.Acupuncture is becoming a popular complementary and alternative treatment in Western countries and is growing in popularity worldwide. In 1985, Dr M. Smith, the head of the US National Acupuncture Detoxification Association, developed a protocol that consisted of insertion of 5 needles without electrical stimulation bilaterally into the outer ear or auricle at points referred to as sympathetic, shenmen, kidney, lung, and liver. The National Acupuncture Detoxification Association reported that the 5-point auricular acupuncture protocol relieves the withdrawal syndrome, prevents symptoms of craving, and increases the likelihood of a patient participating in a long-term treatment program. Electroacupuncture at the Bei-Shu acupoints has been reported to attenuate the prolonged early-stage withdrawal symptoms following heroin detoxification and to suppress anxiety. Governor Vessel acupoints are commonly used in the treatment of heroin addiction. Laser acupuncture is a noninvasive technique that involves stimulation of traditional acupoints with low-intensity, non-thermal laser irradiation. Laser acupuncture integrates the positive effects of both acupuncture and low-level laser therapy. Laser meridian massage is the application of laser acupuncture that involves stimulation along the meridian with low-intensity, non-thermal laser irradiation. The aim of this study is to investigate the effect of adjuvant laser meridian massage in men with OUD on methadone maintenance treatment according to TCM theory, which advocates treatment of the meridians to ameliorate symptoms of opioid withdrawal.Stimulation of acupoints has been used in heroin addicts since 1972. Dr Wen reported that acupuncture combined with electrical stimulation at 4 body points  and 2 ear points (brainstem and shenmen) relieved the withdrawal syndrome in heroin addicts.22.1The study was approved by our institutional human ethics committee . The protocol was registered with ClinicalTrials.gov (identifier NCT04003077). Written informed consent will be obtained from all study participants. Personal information about potential and enrolled participants will be collected, shared, and maintained in an independent closet in order to protect confidentiality before, during, and after the trial.2.2This case-controlled study is being performed in the Department of Psychiatry and Chinese Medicine at Kaohsiung Chang Gung Memorial Hospital. The study was started in February 2019 and will continue until February 2020. Subjects recruited from our institution's addiction treatment center are being allocated to the experimental group . The study participants will receive 12 sessions of laser meridian massage three times weekly for 4 weeks. An age-matched control group of subjects who do not receive laser meridian massage  will also be enrolled. The study design is shown in Figure 2.3A diagnosis of OUD is confirmed using the diagnostic criteria of the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition. Male subjects aged 20 to 70 years with OUD who have received methadone maintenance treatment for at least 1 month and provided informed consent are being recruited. Psychiatrists assess each prospective participant's eligibility to be enrolled in the study. Subjects with a critical illness or human immunodeficiency virus infection, those on antidepressant or antipsychotic medication, those who have taken Chinese herbs or received acupuncture treatment during the previous 30 days, those who are unsuitable for recruitment in the opinion of the attending physician, and those who are unwilling to provide informed consent are excluded.2.4 Anticipating a 7% dropout rate, a total of 30 study participants will need to be recruited.A sample size of 28 was calculated to be needed based on 2-way repeated-measures analysis of variance with a medium effect size of 0.25, a significance level (α) of 0.05, and a desired power (1−β) of 0.80.2.52; power density, 300 mW/cm2; pulsed wave; and frequencies ; RJ-Laser, Reimers & Janssen GmbH, Waldkirch, Germany). The laser treatment will be applied to the back, including the Bladder meridian (BL11–BL25) and Governor Vessel  scores for heroin craving/refusal to use heroin (0–10 points) in the previous week before and after 2 and 4 weeks of treatment and quality of life using the Short Form-12v2 (SF-12v2) before and after 4 weeks of treatment. The participant's pulse diagnosis and heart rate variability after one single treatment session are also being recorded.A VAS score of 0 for heroin craving indicates no heroin craving and a score of 10 indicates the strongest possible heroin craving. A VAS score of 0 for refusal to use heroin indicates no refusal and a score of 10 indicates total refusal. The SF-12v2 Health Survey is a multipurpose, short-form health instrument containing 12 questions that yields an 8-scale profile of functional health and well-being , as well as two psychometrically-based physical and mental health summary measures and a preference-based health utility index. The PRO CoRE, which is part of the Smart Measurement System suite of products and Optum's upgrade to the QualityMetric Health Outcomes scoring software, will be used to score the SF-12v2 Health Survey.The reasons for patients not completing their follow-up visits or dropping out of the study, such as adverse events/intercurrent illnesses, suboptimal response to therapy, failure to return for follow-up, failure to meet the selection criteria at entry, other protocol violations, and refusal to receive treatment, are recorded, as are potential adverse events.2.7t test and chi-square test will be used to evaluate and compare the baseline patient characteristics between the experimental and control groups. The independent t test and McNemar test will be used to compare the differences between 2 groups. Repeated-measures analysis of variance, generalized estimating equations, and the paired t test will be used for comparisons between the 2 study groups. All analyses will be performed using SPSS for Windows, version 22 . A P value < .05 will be considered statistically significant.The data will be presented as the mean ± standard deviation. The independent 2.8A data monitoring committee (DMC) is not needed because laser meridian massage is a routine and noninvasive intervention.3 The prevalence of heroin use was estimated at 0.23% in the population aged 12 to 64 years in 2014  in Taiwan. Addiction to opiates such as heroin is of great importance because it has such serious sequelae, including high mortality and crime rates. Moreover, impaired cortical plasticity has been detected in heroin addicts, which might account for the addiction scores in these individuals  from the perspective of psychomotor stimulant theory or the spiral theory of addiction. Treatment of OUD includes a set of pharmacologic and psychosocial interventions aimed at reducing or ceasing opioid use, preventing future harms associated with opioid use, and improving quality of life and well-being. Pharmacologic strategies have not been successful in management of withdrawal symptoms in heroin addicts; therefore, it is time to investigate the potential benefits of complementary and alternative medicine, which has a long history in the treatment of disease and few side effects.According to the 2019 report of the United Nations Office on Drug and Crime, the global prevalence of opiate (heroin and opium) use was estimated to be 0.6% in the population aged 15 to 64 years in 2017 (29.2 million); furthermore, the number of past-year opiate users globally was 50% higher than the previously estimated 19.4 million in 2016.,19 High-frequency (100-Hz) electroacupuncture is effective in reducing withdrawal symptoms, and there is a suggestion that dynorphin suppresses symptoms of withdrawal at the spinal level. The effects of 100-Hz electroacupuncture may include activation of brain-derived neurotrophic factor in the ventral tegmental area, while multiple 100-Hz stimulation may have a cumulative effect related to acceleration of dynorphin synthesis and downregulation of cAMP response element-binding protein. Furthermore, electroacupuncture could effectively alleviate symptoms of opioid craving and depression, and transcutaneous electrical stimulation of acupoints may help to improve symptoms of insomnia and anxiety.Acupuncture can exert an antinociceptive effect by accelerating the production and release of opioid peptides in the central nervous system. The antinociceptive effect is frequency-dependent, with low-frequency (2-Hz) stimulation accelerating production of endorphin and encephalin and high-frequency (100-Hz) stimulation upregulating the dynorphin level. Laser acupuncture integrates the positive effects of both acupuncture and low-level laser therapy. Laser meridian massage is the application of laser acupuncture. Selection of the meridians for detoxification has been extensive. The meridians that include most acupoints are the Governor Vessel (23.5%) and Bladder meridian (14.7%). Therefore, laser meridian massage is expected to be an effective treatment for patients with OUD on methadone maintenance treatment.In comparison with traditional acupuncture for obtaining qi, laser acupuncture is not related to somatosensation but has the advantage of being noninvasive and aseptic. Furthermore, no heat is generated during the procedure, making laser acupuncture painless and safe.The authors thank the Biostatistics Center at Kaohsiung Chang Gung Memorial Hospital for developing the statistical analysis protocol that will be used in this research.Conceptualization: Wen-Long Hu, Meng-Chang Tsai.Data curation: Wen-Long Hu.Funding acquisition: Wen-Long Hu.Investigation: Wen-Long Hu, Meng-Chang Tsai, Chun-En Kuo, Chun-Ting Liu, Szu-Ying Wu, Tzu-Chan Wu.Methodology: Wen-Long Hu, Meng-Chang Tsai.Project administration: Wen-Long Hu, Meng-Chang Tsai, Chun-En Kuo, Chun-Ting Liu, Szu-Ying Wu, Tzu-Chan Wu.Resources: Wen-Long Hu, Meng-Chang Tsai.Supervision: Wen-Long Hu, Yu-Chiang Hung.Validation: Wen-Long Hu.Visualization: Wen-Long Hu, Yu-Chiang Hung.Writing – original draft: Wen-Long Hu.Writing – review & editing: Meng-Chang Tsai, Yu-Chiang Hung.Wen-Long Hu orcid: 0000-0001-9549-6202.


PEG10, CLDN1, ASS1, WNT7A, GLDC, and RSAD2) significantly associated with poorer prognosis and 3 genes  significantly associated with better prognosis. Combining with the data of microRNA profiles using microRNA target predicting tools, two significantly dysregulated microRNA-mediated gene expression changes in endometrial adenocarcinoma were identified: downregulated hsa-miR-127-5p with upregulated CSTB and upregulated hsa-miR-218-5p with downregulated HPGD. These findings may contribute important new insights into possible novel diagnostic or therapeutic strategies for endometrial adenocarcinoma.Endometrial cancer is one of the most common cancers in women worldwide, affecting more than 300,000 women annually. Dysregulated gene expression, especially those mediated by microRNAs, play important role in the development and progression of cancer. This study aimed to investigate differentially expressed genes in endometrial adenocarcinoma using next generation sequencing (NGS) and bioinformatics. The gene expression profiles and microRNA profiles of endometrial adenocarcinoma (cancer part) and normal endometrial tissue (non-cancer part) were assessed with NGS. We identified 56 significantly dysregulated genes, including 47 upregulated and 9 downregulated genes, in endometrial adenocarcinoma. Most of these genes were associated with defense response, response to stimulus, and immune system process, and further pathway analysis showed that human papillomavirus infection was the most significant pathway in endometrial adenocarcinoma. In addition, these genes were also associated with decreased cell death and survival as well as increased cellular movement. The analyses using Human Protein Atlas, identified 6 genes ( PTEN mutation, hMLH1 methylation, and hMSH6 mutation are important in atypical hyperplastic change of normal endometrium. Mutations in PTEN, KRAS, and CTNNB1 are associated with malignant change from atypical endometrial hyperplasia to low-grade endometrioid cancer, while P53 mutation plays an important role in advancing low-grade cancer into high-grade one P53 and HER2/neu are associated with non-endometrioid malignant transformation from normal or atrophic endometrium Cancers of the corpus uteri, primarily from the endometrium, rank as the sixth most common neoplasm in women worldwide. The incidence increased from 290,000 in 2008 to over 380,000 in 2018 Traditionally, patients suffered from endometrial cancer have a favorable treatment outcome if diagnosed in the early stage. The overall five-year survival rate of endometrial cancer is 81%, but is only 17% if distal metastasis occurs The development of next-generation sequencing (NGS) technologies provides the capability to rapidly sequence exomes, transcriptomes, and genomes at relatively low cost. The application of this technology to catalog the mutational landscapes of tumor exomes, transcriptomes, and genomes has remarkably accelerated the progress in basic and clinical cancer researches As microRNAs have the ability to repress the expression of protein-coding genes, they might contribute to the pathogenesis of various diseases including cancer Through identifying novel gene expression signature and microRNA-gene interactions in endometrial adenocarcinoma, we may provide new perspectives for the development of novel diagnostic methods, prognostic predicting tools, and therapeutic strategies of endometrial adenocarcinoma. Therefore, in this study, we would like to identify the differentially expressed gene and the potential regulatory mechanisms through microRNAs in endometrial adenocarcinoma with systematic bioinformatics analysis.® Pathway Analysis (IPA), the altered functions and pathways related to the dysregulated genes in endometrial cancer were investigated. In addition, the potential targets of the significantly dysregulated microRNAs were predicted with miRmap, TargetScan, and miRDB, and the potential microRNA-mRNA interactions in endometrial cancer were identified.The flowchart of study design is illustrated in Figure ® Reagent  as per the instruction manual. The purified RNAs were quantified at O.D.260nm with a ND-1000 spectrophotometer  and qualitatively assessed with Bioanalyzer 2100 and RNA 6000 LabChip kit . Library preparation and sequencing were performed in Welgene Biotechnology Company .The expression profiles of microRNAs and mRNAs were examined using NGS as in our previous studies p values were calculated by Cuffdiff with non-grouped samples using the "blind mode”, in which all samples were treated as replicates of a single global "condition" and used to build a model for statistical test q values were the p values adjusted with false discovery rate using the method by Benjamini and Hochberg q-value < 0.05  > 1.3) and > 2-fold changes were considered significantly differentially expressed.For transcriptome sequencing, the Agilent's SureSelect Strand Specific RNA Library Preparation Kit was used to construct the libraries, followed by AMPure XP Beads size selection. The sequence was directly determined using Illumina's sequencing-by-synthesis (SBS) technology. Sequencing data (FASTQ files) were generated by Welgene's pipeline based on Illumina's base-calling program bcl2fastq v2.2.0. After adaptor clipping and sequence quality trimming with Trimmomatics (Ver. 0.36) For small RNA sequencing, samples were prepared using Illumina sample preparation kit as per the TruSeq Small RNA Sample Preparation Guide. The 3' and 5' adaptors were ligated to the RNA, and then reverse transcription and PCR amplification were performed. The cDNA constructs were size-fractionated and purified using a 6% polyacrylamide gel electrophoresis and the bands corresponding to the 18-40 nucleotide RNA fragments (140-155 nucleotide in length with both adapters) were extracted. After sequencing on an Illumina  instrument (75 bp single-end reads), the data was processed with the Illumina software. After trimming and filtering out low-quality data with Trimmomatics http://mirmap.ezlab.org/) is an open-source software library which can provide comprehensive prediction of microRNA targets miRmap (http://www.targetscan.org) is an online database predicting the target of microRNA by searching for the presence of conserved 8mer, 7mer, and 6mer sites matching the seed region of each microRNA TargetScan (http://mirdb.org) provides web-based microRNA-target prediction and functional annotations in five species, including human, mouse, rat, dog, and chicken miRDB (https://string-db.org/) has collected and integrated this information, by consolidating known and predicted protein-protein association data of various organisms The functional interactions between expressed proteins in cells are very important and complicated. STRING database (https://david.ncifcrf.gov/) is a powerful tool for functional classification of genes p value in DAVID database which represents how specifically the genes are involved in a category. In this study, we selected EASE score = 0.1 as the default and defined pathways with a q value  <0.05 as significant.DAVID  is a database software containing large database with detailed and structured findings reviewed by experts, which was derived from thousands of biological, chemical and medical researches http://www.elixir-europe.org) as a European core resource due to its fundamental importance for a wider life science community. In this study, the prognosis-predicting values of the significantly dysregulated genes in endometrial cancer were assessed with the Human Protein Atlas. Based on the FPKM value of each gene, the patients were classified into two groups, low-expression and high-expression, to compare their prognoses. The prognosis  of each group of patients was examined and compared with Kaplan-Meier survival estimators and log-rank tests. To determine the best cut-off FPKM values for grouping the patients, all FPKM values from the 20th to 80th percentiles were used to group the patients, and the cut-off FPKM value that yielded the lowest log-rank p value was selected.The Human Protein Atlas is a Swedish-based program initiated in 2003 with the aim to map all the human proteins in cells, tissues, and organs using integration of various omics technologies, including antibody-based imaging, mass spectrometry-based proteomics, transcriptomics, and systems biology 10 > 1.3 and fold change > 2) were identified    network of 56 nodes and 67 edges (enrichment ) Figure . Most ge8) Table .We then used DAVID to analyze the biological processes, cellular components, and molecular functions associated with the 56 significantly dysregulated genes in endometrial adenocarcinoma Table . The sigUsing IPA, the associated diseases and functions of the 56 significantly dysregulated genes in endometrial adenocarcinoma were investigated Figure . The disPEG10, CLDN1, ASS1, WNT7A, GLDC, and RSAD2) and 3 genes were significantly associated with better prognosis .Using the Human Protein Atlas, the prognosis-predicting values of the 56 significantly dysregulated genes in endometrial cancer were assessed Figure  was obtamicroRNA-mRNA interactions with miRmap score ≥ 97.0, and found 34 possible microRNA-mRNA interactions (including 12 interactions between a downregulated microRNA and an upregulated mRNA and 22 interactions between an upregulated microRNA and a downregulated mRNA) involving 16 mRNAs (9 upregulated and 7 downregulated mRNAs)  were identified. We predicted the potential targets of these microRNAs with miRmap database, selecting the targets in the list of the 56 significantly dysregulated genes in endometrial cancer and the CSTB and upregulated hsa-miR-218-5p with downregulated HPGD.In the current study, significantly dysregulated genes, especially those mediated by dysregulated microRNAs, in endometrial adenocarcinoma were investigated comprehensively using an approach with NGS and bioinformatics. We found 56 significantly dysregulated genes, including 47 upregulated and 9 downregulated genes. Most of these genes were associated with defense response, response to stimulus, and immune system process, suggesting the important association between endometrial adenocarcinoma and immune response. Interestingly, the KEGG pathway analysis in STRING showed that HPV infection was the most significant pathway in endometrial adenocarcinoma, suggesting the possible role of HPV in the carcinogenesis of endometrial cancer. Further analyses using DAVID also implied that endometrial adenocarcinoma might be associated with virus infection. On the other hand, the analyses using IPA found that these 56 genes were associated with decreased cell death and survival as well as increased cellular movement, which were common behaviors of malignant cells. The analyses using Human Protein Atlas, identified 6 genes significantly associated with poorer prognosis and 3 genes significantly associated with better prognosis. Combining with the data of microRNA profiles using microRNA target predicting tools, two significantly dysregulated microRNA-mediated gene expression changes in endometrial adenocarcinoma were found: downregulated hsa-miR-127-5p with upregulated It has been debated whether endometrial cancer is associated HPV, although a strong association between HPV and cervical cancer is well-known. The close anatomical proximity to the cervix has led researchers to study whether HPV has a role in the carcinogenesis of endometrial cancer. However, a systematic review and meta-analysis revealed a pooled prevalence of HPV DNA in endometrial cancer tissue of 10.0%  with large between-study heterogeneity related to the HPV DNA detection methods, and concluded that HPV had a limited or no role in the carcinogenesis of endometrial cancer et al. have also demonstrated downregulation of miR-127 in endometrial cancer using microRNA microarray profiling et al. have shown upregulation of miR-218 in endometrial cancer tissue using quantitative reverse transcription polymerase chain reaction Aberrant expression of various microRNAs in endometrial cancer has been reported CSTB gene encodes cystatin B in humans, which may interact with cathepsin B CSTB expression was noted in the cancer tissue, which might be related to the early stage of the cancer in our patient. Further study should be taken to elucidate the role of CSTB.The HPGD gene encodes 15-hydroxyprostaglandin dehydrogenase, an important enzyme responsible for inactivating prostaglandins and associated eicosanoids via reducing the 15S-hydroxyl group, which have been demonstrated as an important factor of disease-associated pain and inflammation in patients with endometriosis HPGD expression is associated with abnormal prostaglandin metabolism in endometriosis HPGD is associated with various types of cancer, such as bladder cancer, gastrointestinal cancer, breast cancer, and cervical cancer 2 and PGF2α) via 15-hydroxyprostaglandin dehydrogenase was noted in type II endometrial cancer, while low HPGD expression was associated with worse progression-free survival and overall survival HPGD expression in endometrial adenocarcinoma tissue, which might be suppressed by upregulated miR-218-5p.The A major limitation of this study should be specified. This study was conducted mainly using the NGS data of a pair of tissue from a patient. Since it was mainly a single subject study, the findings might not be applied to other patients. However, our study provided insights to understand the pathogenic mechanisms of endometrial adenocarcinoma. After further validation, the potential targets identified in our study might provide scientific basis for developing novel diagnostic and treatment modalities for endometrial cancer.CSTB and upregulated hsa-miR-218-5p with downregulated HPGD. These findings may contribute important new insights into possible novel diagnostic or therapeutic strategies for endometrial cancer.In summary, we identified 56 significantly dysregulated genes in endometrial adenocarcinoma. These genes were involved in defense response, response to stimulus, and immune system process, as well as the pathway associated with HPV infection. In these genes, 6 genes were associated with poorer prognosis and 3 genes were associated with better prognosis. We further found two significantly dysregulated microRNA-mediated gene expression alterations in endometrial adenocarcinoma: downregulated hsa-miR-127-5p with upregulated


Pemphigus foliaceus is an autoimmune skin disease mediated by autoantibodies directed against desmoglein-1 located in the upper epidermal layer. Rituximab, a monoclonal anit-CD20 antibody depleting b-cells, offers an effective treatment possibility for therapy-resistant pemphigus foliaceus. Here, we present the case of 55-year-old man who did not respond sufficiently to conventional treatment with prednisolone, azathioprine, and cyclophosphamide, but underwent almost complete remission after rituximab treatment. The patient relapsed 7 years later, and a repeated course of rituximab infusions led to a partial remission. Pemphigus foliaceus is a rare, autoimmune blistering skin disease with an estimated incidence of < 1 million individuals in the USA and Europe , 3. The Glucocorticoids are the first-line treatment for pemphigus foliaceus. Treatment with other immunosuppressants such as azathioprine, mycophenolate mofetil or methotrexate, is also well-established . Recent A 55-year-old man with a history of progressing skin lesions over the past 8 months visited our department for the first time in spring 2011. The clinical examination revealed multiple erythematous papules and plaques with crusts on his back, chest, face, and scalp (about 40% of body surface area was involved) with no mucosal involvement  and azathioprine (100 mg/day) was started. Topical therapy with clobetasol propionate and chlorhexidine was also initiated. Furthermore, methylprednisolone infusions (750 mg) were administered once a month for 3 months. This treatment did not result in complete remission; thus, methylprednisolone was replaced with dexamethasone (300 mg) and cyclophosphamide infusions (500 mg) once a month. Azathioprine had to be discontinued due to increasing liver enzymes. The treatment with cyclophosphamide and glucocorticoids was discontinued after 5 months without achieving remission. Hence, we next treated the patient with rituximab. Therefore, two rituximab infusions (1 g each) were administered 2 weeks apart leading to a near-complete b-cell depletion in peripheral blood, a decrease in Dsg1 antibody levels (below the detection range), and an almost complete remission of the skin lesions within the next year  or two doses of 1,000 mg rituximab are administered 2 weeks apart (rheumatoid arthritis protocol). In their comprehensive review, Ahmed and Shetty  . Few casOur case report demonstrates that rituximab can be effective in treating therapy-resistant pemphigus and long-lasting remission may be achieved. A low-dose maintenance immunomodulatory treatment after rituximab therapy may prolong the remission stage. In case of a relapse, repeated treatment with rituximab is usually successful. However, this therapy is limited due to the high cost of rituximab and the risk of rare but severe side effects.MK wrote the manuscript. MW contributed to conception of the manuscript and revised it critically.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The purpose of this study is to evaluate the role of pre-miR34a rs72631823 as potential risk factor and/or prognostic marker in patients with triple negative breast cancer.114 samples of DNA from paraffin embedded breast normal tissues of patients with triple negative breast cancer and 124 samples of healthy controls were collected and analyzed for pre-miR34a rs72631823 polymorphism.p = 0.176, Fisher’s exact test). The examined polymorphism was not associated with overall survival at the univariate or multivariate Cox regression analysis .Pre-miR34a rs72631823 A allele was associated with increased TNBC risk both in univariate and multivariate analysis. The number of pre-miR34a rs72631823 AA subjects was very small and the association did not reach significance (Our case-control study suggests that pre-miR34a rs72631823 A allele is associated with increased triple negative breast cancer risk. Triple negative breast cancer (TNBC) represents 15–20% of all invasive breast cancers and is characterized by high biological heterogeneity with increased levels of distal recurrence and poor survival, besides high response rate to chemotherapy that constitutes the only standard of treatment . Single MicroRNAs are small, endogenous, non-coding RNAs of approximately 22 nucleotides that could regulate gene expression at post-transcriptional level by binding with mRNAs. Depending on the complementarity between the seed sequence of a microRNA (region of 6-8 nucleotides in the 5’ end of microRNA) and mainly the 3’untranslated region of the target mRNA molecule, microRNAs could induce degradation or translational repression of their targets , 12. ThrSequence alterations in pre-miRNA molecules seem to affect the levels of the mature microRNA . Pre-miRThis study was designed in order to investigate the role of pre-miR-34a rs72631823 polymorphism as a potential risk factor and/or a prognostic marker in patients with TNBC who have received chemotherapy, via a case-control study of 114 patients with triple negative breast cancer and 124 controls.p = 0.023, MWW) and consumed alcohol more frequently , compared to controls. No significant differences were documented in education, menopausal status, smoking rates between cases and controls. The majority of TNBC cases were T2 (61.4%), node-negative (63.2%), grade 3 (86.9%) carcinomas.Table p = 0.176, Fisher’s exact test). The multivariate analysis, adjusting for age, smoking, alcohol consumption, menopausal status, age at menarche and education, confirmed that Pre-miR34a rs72631823 A allele was associated with increased TNBC risk .Genotype frequencies, unadjusted and adjusted ORs for the examined polymorphisms are provided in Table Subgroup analyses by menopausal status reproduced the findings of the overall analysis. In premenopausal women, the adjusted OR for the allele dose-response model was 5.15 (95%CI: 1.22–21.68). Accordingly, in postmenopausal women the adjusted OR for the allele dose-response model was 2.49 (95%CI: 1.20–5.16).p = 0.413).No significant deviation from HWE was documented for the examined polymorphism (Pearson’s chi2(1) = 0.67, The results of the nested prospective study in cases are shown in Table This study is the first to highlight that pre-miR34a rs72631823 A allele is associated with nearly 3-fold increased risk of TNBC. The association was evident in premenopausal as well as postmenopausal women and persisted after adjustment for various potential confounders, including age, smoking, alcohol consumption, age at menarche and education. On the other hand, pre-miR34a rs72631823 A allele did not seem to alter the overall survival of TNBC.This is the first study that evaluates the role of pre-mir34a rs72631823 polymorphism as a potential risk factor or/and prognostic factor in TNBC. Since the examined polymorphism has been previously investigated only once in a line of pancreatic beta cells, and not in cancer, based on current knowledge our results cannot be compared to other studies. However, these findings seem to agree with previous studies stating that alterations in pre-miRNAs could affect the expression levels of genes involved in oncogenesis. The association trend between pre-mir34a rs72631823 and TNBC is in accordance with the studies of Morales S et al  and Li MSince 2005, when Iorio et al  presenteAn important element of validity of the present study is the absence of deviation of allele frequencies in controls for pre-mir34a rs72631823 concerning HWE. Discrepancies on HWE might indicate incorrect or guided sample selection by the research team, genotyping errors or incongruity of mating. Despite the originality, limitations of this case-control study should be reported. First, more studies with increased sample size should be performed to overcome the obstacle of the small number of homozygous pre-mir34a rs72631823 AA subjects. Additionally, in our study, the investigated polymorphism was evaluated in paraffin embedded normal breast tissue, which however is a very common source for DNA extraction in the studies regarding microRNAs.In conclusion, according to our data it seems that pre-mir34a rs72631823 A allele is associated with increased TNBC risk. Future well designed studies should be performed to elucidate the mechanisms underlying the effect of this SNP in TNBC, across different races.Paraffin embedded breast normal tissues were collected from 114 patients with histologically confirmed TNBC during the period 2000-2014. Operations were conducted at the Gynecology Department, “Alexandra” Hospital, Medical School, University of Athens, Greece and patients received chemotherapy at the Oncology Department, “Alexandra” Hospital, Medical School, University of Athens, Greece. The exclusion criteria were as follows: metastatic disease at diagnosis, no invasive disease, family history of breast cancer , history of prior malignancy and no signed informed consent form. Additional information regarding the histology, tumor size, grade, histological stage, lymph node infiltration, the expression levels of ki67 and p53, disease free survival (DFS) and overall survival (OS) were registered in an electronic database according to the patients’ medical records. Concerning controls, women with negative results for breast cancer on routine mammography test were selected. Controls were matched on age (+– 2 years) with patients and had no history of other malignancy. Cases and controls were Caucasian and lived in the same geographical region . Informed consent was obtained from all individual participants included in the study. This case-control study has been approved by the local Institutional Review Board.DNA from paraffin embedded normal breast tissues of patients and DNA from the blood of healthy controls was isolated using the Nucleopsin Tissue kit . Amplification of the selected sequences was performed with allele-specific PCR (polymerase chain reaction), using two allele-specific reverse primers: RG, 5′ CTTGCTGATTGCTTCCTTAC 3′ for the wild type allele and RA: 5′ CTTGCTGATTGCTTCCTTAT 3′ for the mutant allele, in combination with a common forward primer F: 5′ CACATTTCCTTCTTATCAACAG 3′ in two separate PCR reactions. The 3′-ends of the reverse primers were able to anneal to the regions that differed between the two alleles. The endogenous levels of the corresponding gene products were quantified by ELISA.Data were cross-tabulated by case-control status; Mann-Whitney Wilcoxon test (MWW) and Chi-square test were used for the comparison of demographic, lifestyle and reproductive factors in cases and controls, as appropriate. To analyze the associations between the examined polymorphisms and the risk of TNBC, three separate logistic regression models were evaluated: heterozygous vs. wild type (the most frequent homozygous genotype was set as “wild type”), homozygous vs. wild type and dose-response allele model . Unconditional logistic regression analysis was performed to estimate univariate and multivariate odds ratios (ORs) with 95% confidence intervals (CIs). The multivariate ORs were adjusted for age, smoking, alcohol, menopausal status age at menarche and education. In addition, subgroup analyses for premenopausal and postmenopausal women were conducted. The deviation of allele frequencies in controls from the Hardy-Weinberg Equilibrium (HWE) was examined with the appropriate goodness-of-fit Chi-square test, given that the deviation may denote bias . Univari